New valid spectrofluorimetric method for determination of selected cephalosporins in different pharmaceutical formulations using safranin as fluorophore.

PubMed

Derayea, Sayed M; Ahmed, Hytham M; Abdelmageed, Osama H; Haredy, Ahmed M

2016-01-15

A new validated spectrofluorimetric method has been developed for the determination of some cephalosporins namely; cefepime, cefaclor, cefadroxil, cefpodoxime and cefexime. The method was based on the reaction of these drugs with safranin in slightly alkaline medium (pH 8.0), to form ion-association complexes. The fluorescent products were extracted into chloroform and their fluorescence intensities were measured at 544-565 nm after excitation at 518-524 nm. The reaction conditions influencing the product formation and stability were investigated and optimized. The relative fluorescence intensity was proportional to the drug concentration in the linear ranges of 0.15-1.35, 0.35-1.25, 0.35-1.25, 0.20-1.44 and 0.20-1.25 μg/mL for cefepime, cefaclor, cefadroxil, cefpodoxime proxetil and cefexime, respectively. The detection limits were 40, 100, 100, 60 and 70 ng/mL, respectively. The performance of the developed method was evaluated in terms of Student's t-test and variance ratio F-test to find out the significance of proposed methods over the reference spectrophotometric method. Various pharmaceutical formulations were successfully analyzed using the proposed method and the results were in good agreement with those of the previously reported methods. Copyright © 2015. Published by Elsevier B.V.

New valid spectrofluorimetric method for determination of selected cephalosporins in different pharmaceutical formulations using safranin as fluorophore

NASA Astrophysics Data System (ADS)

Derayea, Sayed M.; Ahmed, Hytham M.; Abdelmageed, Osama H.; Haredy, Ahmed M.

2016-01-01

A new validated spectrofluorimetric method has been developed for the determination of some cephalosporins namely; cefepime, cefaclor, cefadroxil, cefpodoxime and cefexime. The method was based on the reaction of these drugs with safranin in slightly alkaline medium (pH 8.0), to form ion-association complexes. The fluorescent products were extracted into chloroform and their fluorescence intensities were measured at 544-565 nm after excitation at 518-524 nm. The reaction conditions influencing the product formation and stability were investigated and optimized. The relative fluorescence intensity was proportional to the drug concentration in the linear ranges of 0.15-1.35, 0.35-1.25, 0.35-1.25, 0.20-1.44 and 0.20-1.25 μg/mL for cefepime, cefaclor, cefadroxil, cefpodoxime proxetil and cefexime, respectively. The detection limits were 40, 100, 100, 60 and 70 ng/mL, respectively. The performance of the developed method was evaluated in terms of Student's t-test and variance ratio F-test to find out the significance of proposed methods over the reference spectrophotometric method. Various pharmaceutical formulations were successfully analyzed using the proposed method and the results were in good agreement with those of the previously reported methods.

Spectrofluorimetric and spectrophotometric stability-indicating methods for determination of some oxicams using 7-chloro-4-nitrobenz-2-oxa-1,3-diazole (NBD-Cl).

PubMed

Taha, Elham Anwer; Salama, Nahla Nour; Fattah, Laila El-Sayed Abdel

2006-05-01

Two sensitive and selective spectrofluorimetric and spectrophotometric stability-indicating methods have been developed for the determination of some non-steroidal anti-inflammatory oxicam derivatives namely lornoxicam (Lx), tenoxicam (Tx) and meloxicam (Mx) after their complete alkaline hydrolysis. The methods are based on derivatization of alkaline hydrolytic products with 7-chloro-4-nitrobenz-2-oxa-1,3-diazole (NBD-Cl). The products showed an absorption maximum at 460 nm for the three studied drugs and fluorescence emission peak at 535 nm in methanol. The color was stable for at least 48 h. The optimum conditions of the reaction were investigated and it was found that the reaction proceeds quantitatively at pH 8, after heating in a boiling water bath for 30 min. The methods were found to be linear in the ranges of 1-10 microg ml(-1) for Lx and Tx and 0.5-4.0 microg ml(-1) for Mx for spectrophotometric method, while 0.05-1.0 microg ml(-1) for Lx and Tx and 0.025-0.4 microg ml(-1) for Mx for the spectrofluorimetric method. The validity of the methods was assessed according to USP guidelines. Statistical analysis of the results revealed high accuracy and good precision. The suggested procedures could be used for the determination of the above mentioned drugs in pure and dosage forms as well as in the presence of their degradation products.

Spectrofluorimetric methods of stability-indicating assay of certain drugs affecting the cardiovascular system

NASA Astrophysics Data System (ADS)

Moussa, B. A.; Mohamed, M. F.; Youssef, N. F.

2011-01-01

Two stability-indicating spectrofluorimetric methods have been developed for the determination of ezetimibe and olmesartan medoxomil, drugs affecting the cardiovascular system, and validated in the presence of their degradation products. The first method, for ezetimibe, is based on an oxidative coupling reaction of ezetimibe with 3-methylbenzothiazolin-2-one hydrazone hydrochloride in the presence of cerium (IV) ammonium sulfate in an acidic medium. The quenching effect of ezetimibe on the fluorescence of excess cerous ions is measured at the emission wavelength, λem, of 345 nm with the excitation wavelength, λex, of 296 nm. Factors affecting the reaction were carefully studied and optimized. The second method, for olmesartan medoxomil, is based on measuring the native fluorescence intensity of olmesartan medoxomil in methanol at λem = 360 nm with λex = 286 nm. Regression plots revealed good linear relationships in the assay limits of 10-120 and 8-112 g/ml for ezetimibe and olmesartan medoxomil, respectively. The validity of the methods was assessed according to the United States Pharmacopeya guidelines. Statistical analysis of the results exposed good Student's t-test and F-ratio values. The introduced methods were successfully applied to the analysis of ezetimibe and olmesartan medoxomil in drug substances and drug products as well as in the presence of their degradation products.

Spectrofluorimetric assay method for glutathione and glutathione transferase using monobromobimane.

PubMed

Yakubu, S I; Yakasai, I A; Musa, A

2011-06-01

The primary role of glutathione transferase is to defend an organism from toxicities through catalyzing the reaction of glutathione (GSH) with potentially toxic compounds or metabolites to their chemically and biologically inert conjugates. The objective of the study was to develop a simple and sensitive spectrofluorimetric assay method for glutathione transferase using monobromobimane (MBB), a non fluorescent compound with electrophilic site. MBB slowly reacted with glutathione to form fluorescent glutathione conjugate and that the reaction was catalysed by glutathione transferase. Both non-enzymatic and enzymatic reaction products of MBB, in presence of GSH in phosphate buffer (pH 6.5), were measured by following increase of fluorescence at wavelength of 475nm. For validation of the assay method, the kinetic parameters such as the apparent Michaelis-Mente constants and maximum rates of conjugate formation as well as the specific activity of rat hepatic glutathione transferase were determined. The method was found to be sensitive, thus, applied to measure glutathione contents of crude preparation of rat hepatic cytosol fraction.

New Spectrofluorimetric Method with Enhanced Sensitivity for Determination of Paroxetine in Dosage Forms and Plasma

PubMed Central

Darwish, Ibrahim A.; Amer, Sawsan M.; Abdine, Heba H.; Al-Rayes, Lama I.

2008-01-01

New simple spectrofluorimetric method with enhanced sensitivity has been developed and validated for the determination of the antidepressant paroxetine (PXT) in its dosage forms and plasma. The method was based on nucleophilic substitution reaction of PXT with 4-chloro-7-nitrobenzo-2-oxa-1,3-diazole in an alkaline medium (pH 8) to form a highly fluorescent derivative that was measured at 545 nm after excitation at 490 nm. The factors affecting the reaction was carefully studied and optimized. The kinetics of the reaction was investigated, and the reaction mechanism was presented. Under the optimized conditions, linear relationship with good correlation coefficient (0.9993) was found between the fluorescence intensity and PXT concentration in the range of 80–800 ng ml−1. The limits of detection and quantitation for the method were 25 and 77 ng ml−1, respectively. The precision of the method was satisfactory; the values of relative standard deviations did not exceed 3%. The proposed method was successfully applied to the determination of PXT in its pharmaceutical tablets with good accuracy; the recovery values were 100.2 ± 1.61%. The results obtained by the proposed method were comparable with those obtained by the official method. The proposed method is superior to the previously reported spectrofluorimetric method for determination of PXT in terms of its higher sensitivity and wider linear range. The high sensitivity of the method allowed its successful application to the analysis of PXT in spiked human plasma. The proposed method is practical and valuable for its routine application in quality control and clinical laboratories for analysis of PXT. PMID:19609398

Derivatization of labetalol hydrochloride for its spectrofluorimetric and spectrophotometric determination inhuman plasma: Application to stability study

NASA Astrophysics Data System (ADS)

Omar, Mahmoud A.; Derayea, Sayed M.; Abdel-Lateef, Mohamed A.; El Hamd, Mohamed A.

2018-02-01

Two simple, selective and accurate methods were developed for the determination of Labetalol hydrochloride in pure form and pharmaceutical tablets. Both methods are based on derivatization of the studied drug with 4-chloro-7-nitrobenzo-2-oxa-1,3-diazole (NBDsbnd Cl) in alkaline medium (pH 7.5).The reaction product was measured spectrofluorimetrically at 540 nm after excitation at 476 nm (method I) or spectrophotometrically at 480 nm (method II). The calibration graphs were rectilinear over the concentration ranges of 0.10-2.0 and 1.0-11.0 μg mL- 1 for methods I and II, respectively. The proposed methods were successfully applied to the analysis of commercial tablets without interference from common excipients. Furthermore, the spectrofluorimetric method was utilized for the in vitro determination of labetalol in spiked human plasma, with a percent mean recovery (n = 3) of 97.80 ± 1.29%. Moreover, the spectrofluorimetric method was extended to examine the stability study of LBT under different stress conditions such as alkaline, acidic, oxidative, photolytic and a thermal degradation.

Derivatization of labetalol hydrochloride for its spectrofluorimetric and spectrophotometric determination inhuman plasma: Application to stability study.

PubMed

Omar, Mahmoud A; Derayea, Sayed M; Abdel-Lateef, Mohamed A; El Hamd, Mohamed A

2018-02-05

Two simple, selective and accurate methods were developed for the determination of Labetalol hydrochloride in pure form and pharmaceutical tablets. Both methods are based on derivatization of the studied drug with 4-chloro-7-nitrobenzo-2-oxa-1,3-diazole (NBDCl) in alkaline medium (pH7.5).The reaction product was measured spectrofluorimetrically at 540nm after excitation at 476nm (method I) or spectrophotometrically at 480nm (method II). The calibration graphs were rectilinear over the concentration ranges of 0.10-2.0 and 1.0-11.0μgmL -1 for methods I and II, respectively. The proposed methods were successfully applied to the analysis of commercial tablets without interference from common excipients. Furthermore, the spectrofluorimetric method was utilized for the in vitro determination of labetalol in spiked human plasma, with a percent mean recovery (n=3) of 97.80±1.29%. Moreover, the spectrofluorimetric method was extended to examine the stability study of LBT under different stress conditions such as alkaline, acidic, oxidative, photolytic and a thermal degradation. Copyright © 2017 Elsevier B.V. All rights reserved.

Net analyte signal standard addition method (NASSAM) as a novel spectrofluorimetric and spectrophotometric technique for simultaneous determination, application to assay of melatonin and pyridoxine

NASA Astrophysics Data System (ADS)

Asadpour-Zeynali, Karim; Bastami, Mohammad

2010-02-01

In this work a new modification of the standard addition method called "net analyte signal standard addition method (NASSAM)" is presented for the simultaneous spectrofluorimetric and spectrophotometric analysis. The proposed method combines the advantages of standard addition method with those of net analyte signal concept. The method can be applied for the determination of analyte in the presence of known interferents. The accuracy of the predictions against H-point standard addition method is not dependent on the shape of the analyte and interferent spectra. The method was successfully applied to simultaneous spectrofluorimetric and spectrophotometric determination of pyridoxine (PY) and melatonin (MT) in synthetic mixtures and in a pharmaceutical formulation.

Development and Validation of Spectrofluorimetric Method for Determination of Biotin in Bulk and Pharmaceutical Preparations via its Oxidation with Cerium (IV)

PubMed Central

Walash, M. I.; Rizk, M.; Sheribah, Z. A.; Salim, M. M.

2010-01-01

A simple and sensitive spectrofluorimetric method was developed for the determination of biotin in pure form and in pharmaceutical preparations. The proposed method is based on the oxidation of the drug with cerium (IV) ammonium sulfate in acidic medium. The fluorescence of the produced Cerium (III) was measured at 365 nm after excitation at 255 nm. The different experimental parameters affecting the development and stability of the reaction were carefully studied and optimized. The method is applicable over the concentration range of 30-120 ng/mL with correlation coefficient of 0.9998. The detection limit (LOD) of biotin was 2.41 ng/mL while quantitation limit (LOQ) was 7.29 ng/mL. The proposed procedure was successfully applied for the determination of biotin in pharmaceutical preparations with mean recoveries of 99.55 ± 0.83 and 101.67 ± 1.53 for biotin ampoules and capsules, respectively. The results obtained were in good agreement with those obtained using the official method. PMID:23675202

Sperm viability assessment in marine invertebrates by fluorescent staining and spectrofluorimetry: A promising tool for assessing marine pollution impact.

PubMed

Gallo, Alessandra; Boni, Raffaele; Tosti, Elisabetta

2018-01-01

The viability of spermatozoa is a crucial parameter to evaluate their quality that is an important issue in ecotoxicological studies. Here, a new method has been developed to rapidly determine the viability of spermatozoa in three marine invertebrates: the ascidian Ciona intestinalis, the sea urchin Paracentrotus lividus and the mollusc Mytilus galloprovincialis. This method employed the dual DNA fluorescent staining coupled with spectrofluorimetric analysis. The dual fluorescent staining used the SYBR-14 stained live spermatozoa and propidium iodide stained degenerated cells that had lost membrane integrity. Stain uptake was assessed by confocal microscopy and then the percentage of live and dead spermatozoa was quantified by spectrofluorimetric analysis. The microscopic examination revealed three populations of spermatozoa: living-SYBR-14 stained, dead-PI stained, and dying-doubly stained spermatozoa. The fluorescence emission peak values recorded in a spectrofluorimeter provide the portion of live and dead spermatozoa showing a significant negative correlation. The stain combination was further validated using known ratios of live and dead spermatozoa. The present study demonstrated that the dual DNA staining with SYBR-14 and propidium iodide was effective in assessing viability of spermatozoa in marine invertebrates and that spectrofluorimetric analysis can be successfully employed to evaluate the percentage of live and dead spermatozoa. The method develop herein is simple, accurate, rapid, sensitive, and cost-effective, so it could be a useful tool by which marine pollutants may be screened for spermiotoxicity. Copyright © 2017 Elsevier Inc. All rights reserved.

Spectrofluorimetric determination of tobramycin in human serum and pharmaceutical preparations by derivatization with fluorescamine.

PubMed

Tekkeli, Serife Evrim Kepekci; Önal, Armağan; Sağırlı, A Olcay

2014-02-01

A simple, sensitive and selective spectrofluorimetric method has been developed for the determination of tobramycin (TOB) in human serum and pharmaceutical preparations. The method is based on the reaction between the primary amino group of TOB and fluorescamine in borate buffer, pH 8.5, to give a highly fluorescent derivative which is measured at 469 nm after excitation at 388 nm. The fluorescence intensity was directly proportional to the concentration over the range 300-1500 ng/mL, with a limit of detection of 65 ng/mL and limit of quantitation of 215 ng/mL. All variables were investigated to optimize the reaction conditions. The method was validated according to International Conference on Harmonization guidelines in terms of specificity, linearity, limit of detection, limit of quantification, accuracy, precision and robustness. Good recoveries were obtained ranging from 97.4 to 100.64%, indicating that no interference was observed from concomitants usually present in pharmaceutical dosage forms. The method was successfully, applied for the analysis of the drug substance in its pharmaceutical preparations and spiked serum samples. Copyright © 2013 John Wiley & Sons, Ltd.

Study of stability of methotrexate in acidic solution spectrofluorimetric determination of methotrexate in pharmaceutical preparations through acid-catalyzed degradation reaction.

PubMed

Sabry, Suzy M; Abdel-Hady, M; Elsayed, M; Fahmy, Osama T; Maher, Hadir M

2003-07-14

Study of the degradation reaction of methotrexate (MTX) in acidic solution was carried out. Optimization of the experimental parameters of MTX acid hydrolysis was investigated. Spectrofluorimetric method for determination of MTX through measurement of its acid-degradation product, 4-amino-4-deoxy-10-methylpteroic acid (AMP), was developed. Stability of the standard solution of MTX prepared in sulfuric acid was discussed in the view of accelerated stability analysis. Two other comparative spectroflourimetric methods based on measuring the fluorescence intensities from either a condensation reaction with acetylacetone-formaldehyde (Hantzsch reaction) or a reaction with fluorescamine were also described. Beer's law validation, accuracy, precision, limits of detection, limits of quantification, and other aspects of analytical merit are presented in the text. The proposed methods were successfully applied for the analysis of MTX in pure drug and tablets dosage form. The sensitivity of the developed methods was favorable, so it was possible to be adopted for determination of MTX in plasma samples for routine use in high-dose MTX therapy.

Development and Validation of highly Sensitive Stability Indicating Spectrofluorimetric Method for Determination of Amlodipine in Pharmaceutical Preparations and Human Plasma.

PubMed

Mohamed, Abdel-Maaboud I; Omar, Mahmoud A; Hammad, Mohamed A; Mohamed, Abobakr A

2016-11-01

A highly sensitive and simple spectrofluorimetric method was developed for the determination of Amlodipine besylate (AML) in its pharmaceutical formulations and spiked human plasma. The proposed method is based on the investigation of the fluorescence spectral behaviour of AML in Tween-80 micellar system. In aqueous solution, the fluorescence intensity of AML was greatly enhanced (160 %) in the presence of Tween-80. The fluorescence intensity was measured at 427 nm after excitation at 385 nm. The fluorescence-concentration plot was rectilinear over the concentration range 0.1-4.0 μg/ml, with lower detection limit of 0.03 μg/ml. The suggested method was successfully applied for the analysis of AML in its commercial tablets alone or in combination with either Atorvastatin or Valsartan. The application of the proposed method was extended to the assay of AML in spiked human plasma and stability studies of AML after exposure to different forced degradation conditions, such as acidic, alkaline, photo- and oxidative conditions, according to ICH guidelines. The results were statistically compared to those obtained by comparison methods and were found to be in good agreement.

Selective spectrofluorimetric method for determination of Lisinopril in pharmaceutical preparations and in presence of hydrochlorothiazide: Application to content uniformity testing.

PubMed

Derayea, Sayed M; Badr El-Din, Khalid M; Mohammed, Fatma F

2017-12-01

A novel sensitive and cost-effective spectrofluorimetric method has been developed and validated for determination of lisinopril (an angiotensin converting enzyme inhibitor) in its pure form and pharmaceutical preparations. The method is based on the reaction of the drug with ninhydrin and phenylacetaldehyde in buffered medium (pH 7.0) to form a highly fluorescent product measured at 460 nm after excitation at 390 nm. Different experimental parameters were optimized and calibration curve was constructed. The fluorescence-concentration relationship was linear in the range of 0.15-4.0 μg mL -1 . The calculated Limit of detection (LOD) and Limit of quantitation (LOQ) were 0.04 and 0.12 μg mL -1 , respectively. The method was successfully applied for the analysis of pharmaceutical preparations containing the studied drug either alone or co-formulated with hydrochlorothiazide. The obtained results were in agreement with those of the reported method in respect to accuracy and precession. Moreover, the method was applied content uniformity testing according to United States Pharmacopeia (USP) guidelines. Copyright © 2017 John Wiley & Sons, Ltd.

Development of spectrofluorimetric method for determination of certain aminoglycoside drugs in dosage forms and human plasma through condensation with ninhydrin and phenyl acetaldehyde.

PubMed

Omar, Mahmoud A; Hammad, Mohamed A; Nagy, Dalia M; Aly, Alshymaa A

2015-02-05

A simple and sensitive spectrofluorimetric method has been developed and validated for determination of amikacin sulfate, neomycin sulfate and tobramycin in pure forms, pharmaceutical formulations and human plasma. The method was based on condensation reaction of cited drugs with ninhydrin and phenylacetaldehyde in buffered medium (pH 6) resulting in formation of fluorescent products which exhibit excitation and emission maxima at 395 and 470nm, respectively. The different experimental parameters affecting the development and stability of the reaction products were carefully studied and optimized. The calibration plots were constructed with good correlation coefficients (0.9993 for tobramycin and 0.9996 for both neomycin and amikacin). The proposed method was successfully applied for the analysis of cited drugs in dosage forms with high accuracy (98.33-101.7)±(0.80-1.26)%. The results show an excellent agreement with the reference method, indicating no significant difference in accuracy and precision. Due to its high sensitivity, the proposed method was applied successfully for determination of amikacin in real human plasma. Copyright © 2014 Elsevier B.V. All rights reserved.

Studying the association complex formation of atomoxetine and fluvoxamine with eosin Y and its application in their fluorimetric determination

NASA Astrophysics Data System (ADS)

Derayea, Sayed M.; Omar, Mahmoud A.; Abu-hassan, Ahmed A.

2018-03-01

A simple, sensitive and non-extractive spectrofluorimetric method has been developed and validated for the determination of two psychoanaleptic drugs, atomoxetine and fluvoxamine, in pure forms and pharmaceutical dosage forms. The proposed method is based on the formation of binary complexes between eosin Y and the studied drugs in the presence of a Teorell-Stenhagen buffer. The quenching of the native fluorescence of eosin Y due to complex formation with the studied drugs was measured spectrofluorimetrically at 545 nm after excitation at 302 nm. At the optimum reaction conditions, the fluorescence quenching values (ΔF) and concentrations were rectilinear over the concentration ranges of 0.2-2.2 and 0.3-2.2 µg ml-1 for atomoxetine and fluvoxamine, respectively. The developed method was successfully applied for the determination of the studied drugs in their pharmaceutical formulations with average percentage recoveries of 100.13 ± 0.66 and 99.69 ± 0.44 for atomoxetine and fluvoxamine, respectively (n = 5), without interference from common excipients.

Studying the association complex formation of atomoxetine and fluvoxamine with eosin Y and its application in their fluorimetric determination

PubMed Central

Omar, Mahmoud A.; Abu-hassan, Ahmed A.

2018-01-01

A simple, sensitive and non-extractive spectrofluorimetric method has been developed and validated for the determination of two psychoanaleptic drugs, atomoxetine and fluvoxamine, in pure forms and pharmaceutical dosage forms. The proposed method is based on the formation of binary complexes between eosin Y and the studied drugs in the presence of a Teorell–Stenhagen buffer. The quenching of the native fluorescence of eosin Y due to complex formation with the studied drugs was measured spectrofluorimetrically at 545 nm after excitation at 302 nm. At the optimum reaction conditions, the fluorescence quenching values (ΔF) and concentrations were rectilinear over the concentration ranges of 0.2–2.2 and 0.3–2.2 µg ml−1 for atomoxetine and fluvoxamine, respectively. The developed method was successfully applied for the determination of the studied drugs in their pharmaceutical formulations with average percentage recoveries of 100.13 ± 0.66 and 99.69 ± 0.44 for atomoxetine and fluvoxamine, respectively (n = 5), without interference from common excipients. PMID:29657744

An efficient spectrofluorimetric method adopts doxazosin, terazosin and alfuzosin coupling with orthophthalaldehyde: Application in human plasma

NASA Astrophysics Data System (ADS)

Omar, Mahmoud A.; Mohamed, Abdel-Maaboud I.; Derayea, Sayed M.; Hammad, Mohamed A.; Mohamed, Abobakr A.

2018-04-01

A new, selective and sensitive spectrofluorimetric method was designed for the quantitation of doxazosin (DOX), terazosin (TER) and alfuzosin (ALF) in their dosage forms and human plasma. The method adopts efficient derivatization of the studied drugs with ortho-phthalaldehyde (OPA), in the presence of 2-mercaptoethanol in borate buffer (pH 9.7) to generate a highly fluorescent isoindole derivatives, which can strongly enhance the fluorescence intensities of the studied drugs, allowing their sensitive determination at 430 nm after excitation at 337 nm. The fluorescence-concentration plots were rectilinear over the ranges (10.0-400.0) ng/mL. Detection and quantification limits were found to be (0.52-3.88) and (1.59-11.76) ng/mL, respectively. The proposed method was validated according to ICH guidelines, and successfully applied for the determination of pharmaceutical preparations of the studied drugs. Moreover, the high sensitivity of the proposed method permits its successful application to the analysis of the studied drugs in spiked human plasma with % recovery (96.12 ± 1.34-100.66 ± 0.57, n = 3). A proposal for the reaction mechanism was presented.

A validated spectrofluorimetric method for the determination of citalopram in bulk and pharmaceutical preparations based on the measurement of the silver nanoparticles-enhanced fluorescence of citalopram/terbium complexes.

PubMed

Khan, Muhammad Naeem; Shah, Jasmin; Jan, Muhammad Rasul; Lee, Sang Hak

2013-01-01

A simple, sensitive, and accurate spectrofluorimetric method was developed for the determination of citalopram in bulk and pharmaceutical preparations. The method is based on the enhancement of the weak fluorescence signal (FL) of the Tb (III)-citalopram system in the presence of silver nanoparticles. Fluorescence intensities were measured at 555 nm after excitation at 281 nm. Prepared silver nanoparticles (AgNPs) were characterized by UV-Visible spectra and transmission electron microscopy (TEM). Various factors affecting the formation of citalopram-Tb (III)-AgNPs complexes were studied and optimized. The fluorescence intensity versus concentration plot was linear over the range 0.02-14 μg mL(-1), with an excellent correlation coefficient of 0.9978. The limit of detection (LOD) and limit of quantification (LOQ) were found to be 7.15 × 10(-6) μg mL(-1) and 2.38 × 10(-5) μg mL(-1) respectively. The proposed method was found to have good reproducibility with a relative standard deviation of 3.66% (n = 6). The interference effects of common excipients found in pharmaceutical preparations were studied. The developed method was validated statistically by performing recoveries studies and successfully applied for the assay of citalopram in bulk powder and pharmaceutical preparations. Percent recoveries were found to range from 98.98% to 100.97% for bulk powder and from 96.57% to 101.77% for pharmaceutical preparations.

Derivative emission spectrofluorimetry for the simultaneous determination of guaifenesin and phenylephrine hydrochloride in pharmaceutical tablets.

PubMed

Maher, Hadir M; Alshehri, Mona M; Al-taweel, Shorog M

2015-05-01

Rapid, simple and sensitive derivative emission spectrofluorimetric methods have been developed for the simultaneous analysis of binary mixtures of guaifenesin (GUA) and phenylephrine hydrochloride (PHE). The methods are based upon measurement of the native fluorescence intensity of the two drugs at λex = 275 nm in methanolic solutions, followed by differentiation using first (D1) and second (D2) derivative techniques. The derivative fluorescence intensity-concentration plots were rectilinear over a range of 0.1-2 µg/mL for both GUA and PHE. The limits of detection were 0.027 (D1, GUA), 0.025 (D2, GUA), 0.031 (D1, PHE) and 0.033 (D2, PHE) µg/mL and limits of quantitation were 0.089 (D1, GUA), 0.083 (D2, GUA), 0.095 (D1, PHE) and 0.097 (D2, PHE) µg/mL. The proposed derivative emission spectrofluorimetric methods (D1 and D2) were successfully applied for the determination of the two compounds in binary mixtures and tablets with high precision and accuracy. The proposed methods were fully validated as per ICH guidelines. Copyright © 2014 John Wiley & Sons, Ltd.

New spectrofluorimetric and spectrophotometric methods for the determination of the analgesic drug, nalbuphine in pharmaceutical and biological fluids.

PubMed

El-Didamony, Akram M; Ali, Ismail I

2013-01-01

We describe the first studies of a simple and sensitive spectrofluorimetric and spectrophotometric methods for the analysis of nalbuphine (NLB) in dosage form and biological fluids. The spectrofluorimetric method was based on the oxidation of NLB with Ce(IV) to produce Ce(III) and its fluorescence was monitored at 352 nm after excitation at 250 nm. The spectrophotometric method involves addition of a known excess of Ce(IV) to NLB in acid medium, followed by determination of residual Ce(IV) by reacting with a fixed amount of methyl orange and measuring absorbance at 510 nm. In both methods, the amount of Ce(IV) reacted corresponds to the amount of NLB and measured fluorescence or absorbance were found to increase linearly with the concentration of NLB, which are corroborated by correlation coefficients of 0.9997 and 0.9999 for spectrofluorimetric and spectrophotometric methods, respectively. Different variables affecting the reaction conditions such as concentrations of Ce(IV), type and concentration of acid medium, reaction time, temperature, and diluting solvents were carefully studied and optimized. The accuracy and precision of the methods were evaluated on intra-day and inter-day basis. The proposed methods were successfully applied for the determination of NLB in pharmaceutical formulation and biological samples with good recoveries. Copyright © 2012 John Wiley & Sons, Ltd.

Spectrofluorimetric determination of thioridazine and flupentixol in dosage forms; application to content uniformity test.

PubMed

Attia, Tamer Z; Omar, Mahmoud A

2016-08-01

A reliable, sensitive, cheap and non-extractive spectrofluorimetric method has been developed and validated for determination of thioridazine and flupentixol based on ternary complex formation with eosin and lead(II) in the presence of methylcellulose as surfactant at pH 3.2. Under the optimum conditions, the quantitative quenching effect of investigated drugs on the native fluorescence of eosin has been investigated. The quenching of the eosin fluorescence was measured at 517 nm after excitation at 462 nm. The different experimental parameters affecting the development and stability of the reaction products were carefully studied and optimized, and the results were satisfactory. The calibration plots were constructed over the range of 0.5-3.0 µg mL(-1) . The developed method was successfully applied for determination of investigated drugs in commercial tablets without interference from common excipients. It was further applied for content uniformity testing of flupentixol in its tablets. Statistical comparisons of the results with those of the reference methods revealed excellent agreement and indicated no significant difference in accuracy and precision. Copyright © 2015 John Wiley & Sons, Ltd. Copyright © 2015 John Wiley & Sons, Ltd.

Determination of gallium at trace levels using a spectrofluorimetric method in synthetic U-Ga and Ga-As solutions.

PubMed

Kara, Derya; Fisher, Andrew; Foulkes, Mike; Hill, Steve J

2010-01-01

A simple, easy to use and selective spectrofluorimetric method for the determination of trace levels of gallium has been developed. A new Schiff base, N-o-vanillidine-2-amino-p-cresol (OVAC) was synthesized and its fluorescence activity with gallium investigated. Based on this chelation reaction, a spectrofluorimetric method has been developed for the determination of gallium in synthetically prepared Ga-U and Ga-As samples buffered at pH 4.0 using acetic acid-sodium acetate. The chelation reaction between Ga(III) and N-o-vanillidine-2-amino-p-cresol was very fast, requiring only 30min at room temperature to complex completely. The limit of detection (LOD) (3sigma) for Ga(III) was 7.17 nM (0.50 microgL(-1)), determined from the analysis of 11 different solutions of 20 microg L(-1) Ga(III). Copyright 2009 Elsevier B.V. All rights reserved.

Rapid and sensitive spectrofluorimetric determination of enrofloxacin, levofloxacin and ofloxacin with 2,3,5,6-tetrachloro- p-benzoquinone

NASA Astrophysics Data System (ADS)

Ulu, Sevgi Tatar

2009-06-01

A highly sensitive spectrofluorimetric method was developed for the first time, for the analysis of three fluoroquinolones (FQ) antibacterials, namely enrofloxacin (ENR), levofloxacin (LEV) and ofloxacin (OFL) in pharmaceutical preparations through charge transfer (CT) complex formation with 2,3,5,6-tetrachloro- p-benzoquinone (chloranil,CLA). At the optimum reaction conditions, the FQ-CLA complexes showed excitation maxima ranging from 359 to 363 nm and emission maxima ranging from 442 to 488 nm. Rectilinear calibration graphs were obtained in the concentration range of 50-1000, 50-1000 and 25-500 ng mL -1 for ENR, LEV and OFL, respectively. The detection limit was found to be 17 ng mL -1 for ENR, 17 ng mL -1 for LEV, 8 ng mL -1 for OFL, respectively. Excipients used as additive in commercial formulations did not interfere in the analysis. The method was validated according to the ICH guidelines with respect to specificity, linearity, accuracy, precision and robustness. The proposed method was successfully applied to the analysis of pharmaceutical preparations. The results obtained were in good agreement with those obtained using the official method; no significant difference in the accuracy and precision as revealed by the accepted values of t- and F-tests, respectively.

First derivative spectrofluorimetric determination of zopiclone and its degradation product, 2-amino-5-chloropyridine, in pharmaceutical formulations with preliminary tool in biological fluids for clinical evidence of zopiclone intake

NASA Astrophysics Data System (ADS)

Al-Attas, Amirah S.; Nasr, Jenny Jeehan; Shalan, Shereen; Belal, Fathalla

2017-06-01

A simple, fast, sensitive and stability-indicating derivative spectrofluorimetric method is presented for the assay of zopiclone (ZOP), a drug with hypnotic effect, and its main degradation product and major contaminant, 2-amino-5-chloropyridine (ACP). The method is based on measuring the inherent fluorescence intensity of both drugs at λex = 300 nm in methanol, then differentiation using D1 (first derivative technique). The developed method was found to be rectilinear over a range of 0.2-4 μg/mL of ZOP and 4-100 ng/mL of ACP. The limits of detection were 0.05 μg/mL of ZOP and 0.2 ng/mL of ACP with the limit of quantitation of 0.17 μg/mL of ZOP and 0.7 ng/mL of ACP. The outcoming results of the proposed method were compared to those obtained by a reference method showing no significant statistical difference between them concerning precision and accuracy. Additionally, the developed method was applied for detecting ACP in spiked human urine and plasma specimens as a tool of clinical evidence of zopiclone intake that can be easily implemented in forensic laboratories. The proposed method was validated as per ICH guidelines.

A simple and sensitive spectrofluorimetric method for analysis of some nitrofuran drugs in pharmaceutical preparations.

PubMed

Belal, Tarek Saied

2008-09-01

A simple, rapid, selective and sensitive spectrofluorimetric method was described for the analysis of three nitrofuran drugs, namely, nifuroxazide (NX), nitrofurantoin (NT) and nitrofurazone (NZ). The method involved the alkaline hydrolysis of the studied drugs by warming with 0.1 M sodium hydroxide solution then dilution with distilled water for NX or 2-propanol for NT and NZ. The formed fluorophores were measured at 465 nm (lambda (Ex) 265 nm), 458 nm (lambda (Ex) 245 nm) and 445 nm (lambda (Ex) 245 nm) for NX, NT and NZ, respectively. The reaction pathway was discussed and the structures of the fluorescent products were proposed. The different experimental parameters were studied and optimized. Regression analysis showed good correlation between fluorescence intensity and concentration over the ranges 0.08-1.00, 0.02-0.24 and 0.004-0.050 microg ml(-1) for NX, NT and NZ, respectively. The limits of detection of the method were 8.0, 1.9 and 0.3 ng ml(-1) for NX, NT and NZ, respectively. The proposed method was validated in terms of accuracy, precision and specificity, and it was successfully applied for the assay of the three nitrofurans in their different dosage forms. No interference was observed from common pharmaceutical adjuvants. The results were favorably compared with those obtained by reference spectrophotometric methods.

Spectrofluorimetric method for the determination of sulpiride in pharmaceutical preparations and human plasma through derivatization with 2-cyanoacetamide.

PubMed

Shah, Jasmin; Jan, M Rasul; Khan, M Naeem; Shah, Sultan

2013-01-01

A sensitive and accurate spectrofluorimetric method has been developed for the determination of sulpiride in pharmaceutical preparations and human plasma. The developed method is based on the derivatization reaction of 2-cyanoacetamide with sulpiride in 30% ammonical solution. The fluorescent derivatized reaction product exhibited maximum fluorescence intensity at 379 nm after excitation at 330 nm. The optimum conditions for derivatization reactions were studied and the fluorescence intensity versus concentration plot was found to be linear over the concentration range 0.2-20.0 µg/mL with a correlation coefficient of 0.9985. The limit of detection (LOD) and limit of quantification (LOQ) were found to be 0.82 and 2.73 ng/mL, respectively. The proposed method was validated according to ICH guidelines. The effects of common excipients and co-administered drugs were also studied. The accuracy of the method was checked using the standard addition method and percent recoveries were found to be in the range of 99.00-101.25% for pharmaceutical preparations and 97.00-97.80% for spiked human plasma. The method was successfully applied to commercial formulations and the results obtained for the proposed method were compared with a high-performance liquid chromatography reference method and statistically evaluated using the Student's t-test for accuracy and the variance ratio F-test for precision. A reaction pathway was also proposed. Copyright © 2012 John Wiley & Sons, Ltd.

Validated sensitive spectrofluorimetric method for determination of antihistaminic drug azelastine HCl in pure form and in pharmaceutical dosage forms: application to stability study.

PubMed

El-Masry, Amal A; Hammouda, Mohammed E A; El-Wasseef, Dalia R; El-Ashry, Saadia M

2017-03-01

A highly sensitive, simple and rapid spectrofluorimetric method was developed for the determination of azelastine HCl (AZL) in either its pure state or pharmaceutical dosage form. The proposed method was based on measuring the native fluorescence of the studied drug in 0.2 M H 2 SO 4 at λ em  = 364 nm after excitation at λ ex  = 275 nm. Different experimental parameters were studied and optimized carefully to obtain the highest fluorescence intensity. The proposed method showed a linear dependence of the fluorescence intensity on drug concentration over a concentration range of 10-250 ng/mL, with a limit of detection of 1.52 ng/mL and limit of quantitation of 4.61 ng/mL. Moreover, the method was successfully applied to pharmaceutical preparations, with percent recovery values (± SD) of 99.96 (± 0.4) and 100.1 (± 0.52) for nasal spray and eye drops, respectively. The results were in good agreement with those obtained by the comparison method, as revealed by Student's t-test and the variance ratio F-test. The method was extended to study the stability of AZL under stress conditions, where the drug was exposed to neutral, acidic, alkaline, oxidative and photolytic degradation according to International Conference on Harmonization (ICH) guidelines. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

Spectrophotometric, spectrofluorimetric, and densitometric methods for the determination of indapamide.

PubMed

Youssef, Nadia F

2003-01-01

Three sensitive spectrophotometric, spectrofluorimetric, and densitometric methods are described for the determination of indapamide. The first and second methods are based on the oxidative coupling reaction of indapamide with 3-methyl-2-benzothiazolinone hydrazone HCl (MBTH) in the presence of cerium(IV) ammonium sulfate in an acidic medium. The absorbance of the reaction product is measured at the lambdamax, 601 nm. With the same reaction, indapamide is determined by its quenching effect on the fluorescence of excess cerous ions at the emission lambdamax, 350 nm, and the excitation at lambdamax, 300 nm. The reaction conditions were optimized, and Beer's law was obeyed for indapamide at 1.2-9.6 microg/mL with mean recoveries of 99.92 +/- 0.83 and 99.97 +/- 1.11%, respectively. The third method, a stability-indicating densitometric assay, was developed for the determination of indapamide, using toluene-ethyl acetate-glacial acetic acid (69 + 30 + 1, v/v/v) as the developing system and scanning at the lambdamax, 242 nm, in the presence of the degradation product and related substance; for the indapamide concentration range of 0.6-6 microg/spot, the mean recovery was 99.73 +/- 0.71%. The proposed methods were successfully applied to the determination of indapamide in bulk powder and commercial tablets, and the results of the analysis agreed statistically with those obtained with the official method. Furthermore, the methods were validated according to the guidelines of the U.S. Pharmacopeia and also assessed by applying the standard additions technique.

First derivative emission spectrofluorimetric method for the determination of LCZ696, a newly approved FDA supramolecular complex of valsartan and sacubitril in tablets.

PubMed

Ragab, Marwa A A; Galal, Shereen M; Korany, Mohamed A; Ahmed, Aya R

2017-12-01

LCZ696 (sacubitril/valsartan, Entresto™) is a therapy lately approved by United States Food and Drug Administration (US FDA) as a heart failure therapy. It is claimed to decrease the mortality rate and hospitalization for patients with chronic heart failure. This study is considered as the first report to investigate the fluorimetric behavior of sacubitril in addition to pursuing all the different conditions that may affect its fluorescence. Various conditions were studied, for example studying the effects of organized media, solvents and pH, which may affect the fluorescence behavior of sacubitril. For the simultaneous determination of the newly approved supramolecular complex of valsartan (VAL) and sacubitril (SAC) in their tablets, a sensitive and simple first derivative spectrofluorimetric method was developed. The method involved the measurement of native fluorescence at 416 nm and 314 nm (λ ex 249 nm) for VAL and SAC, respectively. The first (D1) derivative technique was applied to the emission data to resolve a partial overlap that appeared in their emission spectra. The proposed method was successfully applied for the assay of the two drugs in their supramolecular complex LCZ696 with no interference from common pharmaceutical additives. International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH) guidelines were followed in order to validate the proposed method. Copyright © 2017 John Wiley & Sons, Ltd.

Application of normal fluorescence and stability-indicating derivative synchronous fluorescence spectroscopy for the determination of gliquidone in presence of its fluorescent alkaline degradation product

NASA Astrophysics Data System (ADS)

El-ghobashy, Mohamed R.; Yehia, Ali M.; Helmy, Aya H.; Youssef, Nadia F.

2018-01-01

Simple, smart and sensitive normal fluorescence and stability-indicating derivative synchronous spectrofluorimetric methods have been developed and validated for the determination of gliquidone in the drug substance and drug product. Normal spectrofluorimetric method of gliquidone was established in methanol at λ excitation 225 nm and λ emission 400 nm in concentration range 0.2-3 μg/ml with LOD equal 0.028. The fluorescence quantum yield of gliquidone was calculated using quinine sulfate as a reference and found to be 0.542. Stability-indicating first and third derivative synchronous fluorescence spectroscopy were successfully utilized to overcome the overlapped spectra in normal fluorescence of gliquidone and its alkaline degradation product. Derivative synchronous methods are based on using the synchronous fluorescence of gliquidone and its degradation product in methanol at Δ λ50 nm. Peak amplitude in the first derivative of synchronous fluorescence spectra was measured at 309 nm where degradation product showed zero-crossing without interference. The peak amplitudes in the third derivative of synchronous fluorescence spectra, peak to trough were measured at 316,329 nm where degradation product showed zero-crossing. The different experimental parameters affecting the normal and synchronous fluorescence intensity of gliquidone were studied and optimized. Moreover, the cited methods have been validated as per ICH guidelines. The peak amplitude-concentration plots of the derivative synchronous fluorescence were linear over the concentration range 0.05-2 μg/ml for gliquidone. Limits of detection were 0.020 and 0.022 in first and third derivative synchronous spectra, respectively. The adopted methods were successfully applied to commercial tablets and the results demonstrated that the derivative synchronous fluorescence spectroscopy is a powerful stability-indicating method, suitable for routine use with a short analysis time. Statistical comparison between the results obtained by normal fluorescence and derivative synchronous methods and the official one using student's t-test and F-ratio showed no significant difference regarding accuracy and precision.

An eco-friendly direct spectrofluorimetric method for the determination of irreversible tyrosine kinase inhibitors, neratinib and pelitinib: application to stability studies.

PubMed

Maher, H M; Alzoman, N Z; Shehata, S M

2017-03-01

A new rapid and simple stability-indicating spectrofluorimetric method has been developed for the determination of two irreversible tyrosine kinase inhibitors (TKIs), neratinib (NER) and pelitinib (PEL). The method is based upon measurement of the native fluorescence intensity of both drugs at λ ex 270 nm in aqueous borate buffer solutions (pH 10.5). The fluorescence intensity recorded at 545 nm (NER) and 465 nm (PEL) were rectilinear over the concentration range of 0.1-10 μg/mL for both drugs with a high correlation coefficient (r > 0.999). The proposed method provided low limits of detection and of quantitation of 0.07, 0.11 μg/mL (NER) and 0.02, 0.05 μg/mL (PEL), respectively. The method was successfully applied for the determination of NER and PEL in bulk powder. The proposed methods were fully validated as per the International Conference on Harmonisation (ICH) guidelines. The application of the method was extended to stability studies of both NER and PEL under different forced-degradation conditions (acidic-induced, base-induced, oxidative, wet heat, and photolytic degradation). Moreover, the kinetics of the base-induced and oxidative degradation of both drugs was investigated and the pseudo-first-order rate constants and half-lives were estimated at different temperatures. Also, an Arrhenius plot was applied to predict the stability behaviour of the two drugs at room temperature. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

Chemometrics-assisted Spectrofluorimetric Determination of Two Co-administered Drugs of Major Interaction, Methotrexate and Aspirin, in Human Urine Following Acid-induced Hydrolysis.

PubMed

Maher, Hadir M; Ragab, Marwa A A; El-Kimary, Eman I

2015-01-01

Methotrexate (MTX) is widely used to treat rheumatoid arthritis (RA), mostly along with non-steroidal anti-inflammatory drugs (NSAIDs), the most common of which is aspirin or acetyl salicylic acid (ASA). Since NSAIDs impair MTX clearance and increase its toxicity, it was necessary to develop a simple and reliable method for the monitoring of MTX levels in urine samples, when coadministered with ASA. The method was based on the spectrofluorimetric measurement of the acid-induced hydrolysis product of MTX, 4-amino-4-deoxy-10-methylpteroic acid (AMP), along with the strongly fluorescent salicylic acid (SA), a product of acid-induced hydrolysis of aspirin and its metabolites in urine. The overlapping emission spectra were resolved using the derivative method (D method). In addition, the corresponding derivative emission spectra were convoluted using discrete Fourier functions, 8-points sin xi polynomials, (D/FF method) for better elimination of interferences. Validation of the developed methods was carried out according to the ICH guidelines. Moreover, the data obtained using derivative and convoluted derivative spectra were treated using the non-parametric Theil's method (NP), compared with the least-squares parametric regression method (LSP). The results treated with Theil's method were more accurate and precise compared with LSP since the former is less affected by the outliers. This work offers the potential of both derivative and convolution using discrete Fourier functions in addition to the effectiveness of using the NP regression analysis of data. The high sensitivity obtained by the proposed methods was promising for measuring low concentration levels of the two drugs in urine samples. These methods were efficiently used to measure the drugs in human urine samples following their co-administration.

Spectrophotometric and spectrofluorimetric determination of indacaterol maleate in pure form and pharmaceutical preparations: application to content uniformity.

PubMed

El-Ashry, S M; El-Wasseef, D R; El-Sherbiny, D T; Salem, Y A

2015-09-01

Two simple, rapid, sensitive and precise spectrophotometric and spectrofluorimetric methods were developed for the determination of indacaterol maleate in bulk powder and capsules. Both methods were based on the direct measurement of the drug in methanol. In the spectrophotometric method (Method I) the absorbance was measured at 259 nm. The absorbance-concentration plot was rectilinear over the range 1.0-10.0 µg mL(-1) with a lower detection limit (LOD) of 0.078 µg mL(-1) and lower quantification limit (LOQ) of 0.238 µg mL(-1). Meanwhile in the spectrofluorimetric method (Method II) the native fluorescence was measured at 358 nm after excitation at 258 nm. The fluorescence-concentration plot was rectilinear over the range of 1.0-40.0 ng mL(-1) with an LOD of 0.075 ng mL(-1) and an LOQ of 0.226 ng mL(-1). The proposed methods were successfully applied to the determination of indacaterol maleate in capsules with average percent recoveries ± RSD% of 99.94 ± 0.96 for Method I and 99.97 ± 0.81 for Method II. In addition, the proposed methods were extended to a content uniformity test according to the United States Pharmacopoeia (USP) guidelines and were accurate, precise for the capsules studied with acceptance value 3.98 for Method I and 2.616 for Method II. Copyright © 2015 John Wiley & Sons, Ltd.

Flow injection determination of aluminium by spectrofluorimetric detection after complexation with N-o-vanillidine-2-amino-p-cresol: the application to natural waters.

PubMed

Kara, Derya; Fisher, Andrew; Hill, Steve J

2008-03-17

An on-line flow injection spectrofluorimetric method for the direct determination of aluminium in water samples is described. The method is based on the reaction of aluminium with N-o-vanillidine-2-amino-p-cresol (OVAC) in acidic medium at pH 4.0 to form a water-soluble complex. The excitation and emission wavelengths were 423.0 and 553.0nm, respectively, at which the OVAC-Al complex gave the maximum fluorescence intensity at pH 4.0 in a 50% methanol-50% water medium at 50 degrees C. An interference from fluoride ions was minimised by the addition of Be(2+). Other ions were found not to interfere at the concentrations likely to be found in natural waters. The proposed methods were validated in terms of linearity, repeatability, detection limit, accuracy and selectivity. Under these conditions, the calibration was linear up to 1000microgL(-1) (r=0.999). The limit of detection (3sigma) for the determination of Al(III) was 0.057microgL(-1) and the precision for multiple determinations of 3ngmL(-1) Al(III) prepared in ultra-pure water was found to be 0.62% (n=10). The Schiff base ligand could be used to determine ultra-trace aluminium from natural waters. Analysis of environmental certified reference materials showed good agreement with the certified values. The procedure was found to be equally applicable to both freshwater and saline solutions, including seawater.

Spectrofluorimetric determination of fluoroquinolones in pharmaceutical preparations.

PubMed

Ulu, Sevgi Tatar

2009-02-01

Simple, rapid and highly sensitive spectrofluorimetric method is presented for the determination of four fluoroquinolone (FQ) drugs, ciprofloxacin, enoxacin, norfloxacin and moxifloxacin in pharmaceutical preparations. Proposed method is based on the derivatization of FQ with 4-chloro-7-nitrobenzofurazan (NBD-Cl) in borate buffer of pH 9.0 to yield a yellow product. The optimum experimental conditions have been studied carefully. Beer's law is obeyed over the concentration range of 23.5-500 ng mL(-1) for ciprofloxacin, 28.5-700 ng mL(-1) for enoxacin, 29.5-800 ng mL(-1) for norfloxacin and 33.5-1000 ng mL(-1) for moxifloxacin using NBD-Cl reagent, respectively. The detection limits were found to be 7.0 ng mL(-1) for ciprofloxacin, 8.5 ng mL(-1) for enoxacin, 9.2 ng mL(-1) for norfloxacin and 9.98 ng mL(-1) for moxifloxacin, respectively. Intra-day and inter-day relative standard deviation and relative mean error values at three different concentrations were determined. The low relative standard deviation values indicate good precision and high recovery values indicate accuracy of the proposed methods. The method is highly sensitive and specific. The results obtained are in good agreement with those obtained by the official and reference method. The results presented in this report show that the applied spectrofluorimetric method is acceptable for the determination of the four FQ in the pharmaceutical preparations. Common excipients used as additives in pharmaceutical preparations do not interfere with the proposed method.

Spectrofluorimetric Method for Estimation of Curcumin in Rat Blood Plasma: Development and Validation

NASA Astrophysics Data System (ADS)

Trivedi, J.; Variya, B.; Gandhi, H.; Rathod, S. P.

2016-01-01

Curcumin is a medicinally important phytoconstituent of curcuminoids. The present study describes development of a simple method for estimation of curcumin in rat plasma. This method involves the use of spectrofluorimetry for evaluation of curcumin at 257 (Ex) and 504 nm (Em). Sample preparation involves only two steps: extraction of curcumin and drying the extract. Following this procedure, the samples are reconstituted with ethyl acetate, and relative fluorescence intensity is measured using a spectrofluorimeter. The method was validated as per CDER guidelines. The linearity of the method was found to be in the range of 100-500 ng/mL with accuracy and precision lying within 2% RSD. The LOD and LOQ were found to be 15.3 and 46.1 ng/mL, respectively. The method was applied for pharmacokinetic evaluation in rats, and AUC, Cmax, and Tmax were found to be 5580 ± 1006 h × ng/mL, 1526 ± 209 ng/mL, and 2.97 ± 0.28 h, respectively, with a plasma half-life of 1.14 ± 0.27 h.

Spectrofluorimetric determination of some water-soluble vitamins.

PubMed

Mohamed, Abdel-Maaboud I; Mohamed, Horria A; Abdel-Latif, Niveen M; Mohamed, Marwa R

2011-01-01

Two simple and sensitive spectrofluorimetric methods were developed for determination of three water-soluble vitamins (B1, B2, and B6) in mixtures in the presence of cyanocobalamin. The first one was for thiamine determination, which depends on the oxidation of thiamine HCl to thiochrome by iodine in an alkaline medium. The method was applied accurately to determine thiamine in binary, ternary, and quaternary mixtures with pyridoxine HCl, riboflavin, and cyanocobalamin without interference. In the second method, riboflavin and pyridoxine HCl were determined fluorimetrically in acetate buffer, pH 6. The three water-soluble vitamins (B1, B2, and B6) were determined spectrofluorimetrically in binary, ternary, and quaternary mixtures in the presence of cyanocobalamin. All variables were studied in order to optimize the reaction conditions. Linear relationship was obeyed for all studied vitamins by the proposed methods at their corresponding lambda(exc) or lambda(em). The linear calibration curves were obtained from 10 to 500 ng/mL; the correlation ranged from 0.9991 to 0.9999. The suggested procedures were applied to the analysis of the investigated vitamins in their laboratory-prepared mixtures and pharmaceutical dosage forms from different manufacturers. The RSD range was 0.46-1.02%, which indicates good precision. No interference was observed from common pharmaceutical additives. Good recoveries (97.6 +/- 0.7-101.2 +/- 0.8%) were obtained. Statistical comparison of the results with reported methods shows excellent agreement and indicates no significant difference in accuracy and precision.

Spectrofluorimetric study of host-guest complexation of ibuprofen with β-cyclodextrin and its analytical application

NASA Astrophysics Data System (ADS)

Manzoori, Jamshid L.; Amjadi, Mohammad

2003-03-01

The characteristics of host-guest complexation between β-cyclodextrin (β-CD) and two forms of ibuprofen (protonated and deprotonated) were investigated by fluorescence spectrometry. 1:1 stoichiometries for both complexes were established and their association constants at different temperatures were calculated by applying a non-linear regression method to the change in the fluorescence of ibuprofen that brought about by the presence of β-CD. The thermodynamic parameters (Δ H, Δ S and Δ G) associated with the inclusion process were also determined. Based on the obtained results, a sensitive spectrofluorimetric method for the determination of ibuprofen was developed with a linear range of 0.1-2 μg ml -1 and a detection limit of 0.03 μg ml -1. The method was applied satisfactorily to the determination of ibuprofen in pharmaceutical preparations.

Spectrofluorimetric study of host-guest complexation of ibuprofen with beta-cyclodextrin and its analytical application.

PubMed

Manzoori, Jamshid L; Amjadi, Mohammad

2003-03-15

The characteristics of host-guest complexation between beta-cyclodextrin (beta-CD) and two forms of ibuprofen (protonated and deprotonated) were investigated by fluorescence spectrometry. 1:1 stoichiometries for both complexes were established and their association constants at different temperatures were calculated by applying a non-linear regression method to the change in the fluorescence of ibuprofen that brought about by the presence of beta-CD. The thermodynamic parameters (deltaH, deltaS and deltaG) associated with the inclusion process were also determined. Based on the obtained results, a sensitive spectrofluorimetric method for the determination of ibuprofen was developed with a linear range of 0.1-2 microg ml(-1) and a detection limit of 0.03 microg ml(-1). The method was applied satisfactorily to the determination of ibuprofen in pharmaceutical preparations. Copyright 2002 Elsevier Science B.V.

A novel spectrofluorimetric method for the assay of pseudoephedrine hydrochloride in pharmaceutical formulations via derivatization with 4-chloro-7-nitrobenzofurazan.

PubMed

El-Didamony, Akram M; Gouda, Ayman A

2011-01-01

A new highly sensitive and specific spectrofluorimetric method has been developed to determine a sympathomimetic drug pseudoephedrine hydrochloride. The present method was based on derivatization with 4-chloro-7-nitrobenzofurazan in phosphate buffer at pH 7.8 to produce a highly fluorescent product which was measured at 532 nm (excitation at 475 nm). Under the optimized conditions a linear relationship and good correlation was found between the fluorescence intensity and pseudoephedrine hydrochloride concentration in the range of 0.5-5 µg mL(-1). The proposed method was successfully applied to the assay of pseudoephedrine hydrochloride in commercial pharmaceutical formulations with good accuracy and precision and without interferences from common additives. Statistical comparison of the results with a well-established method showed excellent agreement and proved that there was no significant difference in the accuracy and precision. The stoichiometry of the reaction was determined and the reaction pathway was postulated. Copyright © 2010 John Wiley & Sons, Ltd.

Determination of the anti-viral drug Ribavirin in dosage forms via micelle-enhanced spectrofluorimetric method.

PubMed

Alarfaj, Nawal A; El-Tohamy, Maha F

2013-01-01

A simple and sensitive spectrofluorimetric method was developed for the determination of Ribavirin in pharmaceutical formulations. The proposed method was based on the fluorescence spectral behaviour of Ribavirin in a sodium dodecyl sulfate (SDS) micellar system. In an aqueous acetate buffer solution of pH 4.0, the fluorescence intensity of Ribavirin was significantly enhanced by about 217% in the presence of SDS. Fluorescence intensity was measured at 396 nm after excitation at 270 nm for Ribavirin. The fluorescence-concentration plot was rectilinear over the range of 0.01-3.0 µg/mL for Ribavirin with a lower detection limit of 5.02 x 10(-3) µg/mL. The method was successfully applied to the analysis of the drug in its commercial capsules. Results were in good agreement with those obtained with the official method. The application of the proposed method was extended to stability studies of Ribavirin after exposure to different forced degradation conditions such as acidic, alkaline, photo and oxidative conditions according to ICH guidelines. Copyright © 2012 John Wiley & Sons, Ltd.

Micelle-enhanced spectrofluorimetric method for determination of cholesterol-reducing drug ezetimibe in dosage forms.

PubMed

Alarfaj, Nawal A; Aly, Fatma A

2012-01-01

A simple and sensitive spectrofluorimetric method was developed for the determination of ezetimibe in its pharmaceutical formulations. The proposed method is based on investigation of the fluorescence spectral behavior of ezetimibe in sodium dodecyl sulfate (SDS) micellar system. In aqueous solution of acetate buffer pH 5.0, the fluorescence intensity of ezetimibe was greatly enhanced, 200% enhancement, in the presence of SDS. The fluorescence intensity of ezetimibe was measured at 380 nm after excitation at 268 nm. The fluorescence-concentration plot was rectilinear over the range of 0.03-3.0 μg/mL with lower detection limit of 3.08 × 10(-3) μg/mL. The method was successfully applied to the analysis of ezetimibe in its commercial tablets; the results were in good agreement with those obtained with the reported method. The application of the proposed method was extended to the stability studies of ezetimibe after exposure to different forced degradation conditions, such as acidic, alkaline, photo and oxidative conditions, according to ICH guidelines.

Spectrofluorimetric determination of DL-N-methyl-3-phenyl-3-[(α,α,α-trifluoro-p-tolyl)oxy]propylamine in pharmaceuticals and chitosan solution.

PubMed

Rodrigues, Máira Regina

2012-02-01

A spectrofluorimetric method for the determination of D,L-N-methyl-3-phenyl-3-[(α,α,α-trifluoro-p-tolyl)oxy]propylamine, fluoxetine (F), in pharmaceuticals was evaluated in the 50.0-500.0 μg ml⁻¹ range. Linearity, sensibility, quantification and detection limit, and precision values are satisfactory. The method does not need pre-treatment and was successfully applied to the determination in pharmaceuticals and chitosan (Ch) solution. Ch has an ability to carry and absorb fat and may eventually be used together with F in slimming diets, and then interactions of Ch-F may occur. This work seeks to study these interactions by monitoring the photophysics of a drug in the presence of Ch. The results warn about the care that must be taken when both compounds are prescribed together.

Sensitive spectrofluorimetric determination of tizanidine in pharmaceutical preparations, human plasma and urine through derivatization with dansyl chloride.

PubMed

Ulu, Sevgi Tatar

2012-01-01

A sensitive spectrofluorimetric method was developed for the determination of tizanidine in human plasma, urine and pharmaceutical preparations. The method is based on reaction of tizanidine with 1-dimethylaminonaphthalene-5-sulphonyl chloride (dansyl chloride) in an alkaline medium to form a highly fluorescent derivative that was measured at 511 nm after excitation at 383 nm. The different experimental parameters affecting the fluorescence intensity of tizanidine was carefully studied and optimized. The fluorescence-concentration plots were rectilinear over the ranges 50-500 and 20-300 ng/mL for plasma and urine, respectively, detection limits of 1.81 and 0.54 ng/mL and quantification limits of 5.43 and 1.62 ng/mL for plasma and urine, respectively. The method presents good performance in terms of linearity, detection and quantification limits, precision, accuracy and specificity. The proposed method was successfully applied for the determination of tizanidine in pharmaceutical preparations. The results obtained were compared with a reference method, using t- and F-tests. Copyright © 2011 John Wiley & Sons, Ltd.

Spectrofluorimetric study of the beta-cyclodextrin-ibuprofen complex and determination of ibuprofen in pharmaceutical preparations and serum.

PubMed

Hergert, L A; Escandar, G M

2003-06-13

The inclusion complexation of ibuprofen in beta-cyclodextrin (beta-CD) has been examined by means of spectrofluorimetry at both acid and alkaline pH. The results suggest that stable 1:1 complexes are formed in both media. The analysis of the pK(a) values for ibuprofen in both the absence and presence of beta-CD (4.12 and 4.66, respectively) suggests that in the inclusion complex the carboxylic group is located outside the cyclodextrin (CD) but interacting with it. Further structural characterization of the complex was carried out by means of am1 semiempiral calculations. Based on the obtained results, a spectrofluorimetric method for the determination of ibuprofen in the presence of beta-CD at 10 degrees C was developed in the range of 4.7-58 mug ml(-1). Better limits of detection (1.6 mug ml(-1)) and quantification (4.7 mug ml(-1)) were obtained in this latter case with respect to those obtained in the absence of beta-CD. The method was satisfactorily applied to the quantification of ibuprofen in pharmaceutical preparations. A novel spectrofluorimetric determination of ibuprofen in the presence of beta-CD was also developed for serum samples at concentration levels between 5 and 70 mug ml(-1). It uses second-order fluorescence excitation-emission matrices coupled to an algorithm based on self-weighted alternating trilinear decomposition (SWATLD), and avoids resorting to separative instrumental analyses.

Highly sensitive spectrofluorimetric determination of trace amounts NADP using Europium ion-doxycycline complex probe

NASA Astrophysics Data System (ADS)

Peng, Qian; Hou, Faju; Jiang, Chongqiu

2006-09-01

A new spectrofluorimetric method was developed for determination of trace amount of Coenzyme II (NADP). Using europium ion-doxycycline (DC) as a fluorescent probe, in the buffer solution of pH 8.44, NADP can remarkably enhance the fluorescence intensity of the Eu 3+-DC complex at λ = 612 nm and the enhanced fluorescence intensity is in proportion to the concentration of NADP. Optimum conditions for the determination of NADP were also investigated. The dynamic range for the determination of NADP is 3.3 × 10 -7 to 6.1 × 10 -6 mol l -1 with detection limit of 6.8 × 10 -8 mol l -1. This method is simple, practical and relatively free interference from coexisting substances and can be successfully applied to determination of NADP in synthetic water samples and in serum samples. Moreover, the enhancement mechanisms of the fluorescence intensity in the Eu 3+-DC system and the Eu 3+-DC-NADP system have been also discussed.

Utility of eosin Y as a complexing reagent for the determination of citalopram hydrobromide in commercial dosage forms by fluorescence spectrophotometry.

PubMed

Azmi, Syed Najmul Hejaz; Al-Fazari, Ahlam; Al-Badaei, Munira; Al-Mahrazi, Ruqiya

2015-12-01

An accurate, selective and sensitive spectrofluorimetric method was developed for the determination of citalopram hydrobromide in commercial dosage forms. The method was based on the formation of a fluorescent ion-pair complex between citalopram hydrobromide and eosin Y in the presence of a disodium hydrogen phosphate/citric acid buffer solution of pH 3.4 that was extractable in dichloromethane. The extracted complex showed fluorescence intensity at λem = 554 nm after excitation at 259 nm. The calibration curve was linear over at concentrations of 2.0-26.0 µg/mL. Under optimized experimental conditions, the proposed method was validated as per ICH guidelines. The effect of common excipients used as additives was tested and the tolerance limit calculated. The limit of detection for the proposed method was 0.121 μg/mL. The proposed method was successfully applied to the determination of citalopram hydrobromide in commercial dosage forms. The results were compared with the reference RP-HPLC method. Copyright © 2015 John Wiley & Sons, Ltd.

Spectrofluorimetric assessment of hydrochlorothiazide using optical sensor nano-composite terbium ion doped in sol-gel matrix.

PubMed

Youssef, A O

2012-05-01

A new, simple, sensitive and selective spectrofluorimetric method for the determination of Hydrochlorothiazide was developed in acetonitrile at pH 6.2. The Hydrochlorothiazide can remarkably enhance the luminescence intensity of the Tb(3+) ion doped in sol-gel matrix at λ(ex) = 370 nm. The intensity of the emission band of Tb(3+) ion doped in sol-gel matrix was increased due to the energy transfer from the triplet excited state of Hydrochlorothiazide to ((5)D(4)) excited energy state of Tb(3) ion. The enhancement of the emission band of Tb(3+) ion doped in sol-gel matrix at ((5)D(4)→(7)F(5)) 545 nm was directly proportion to the concentration of Hydrochlorothiazide with a dynamic ranges of 5.0 × 10(-10)-5.0 × 10(-6) mol L(-1) and detection limit of 2.2 × 10(-11) mol L(-1).

Highly sensitive and selective spectrofluorimetric determination of metoclopramide hydrochloride in pharmaceutical tablets and serum samples using Eu3+ ion doped in sol-gel matrix.

PubMed

Attia, M S; Aboaly, M M

2010-06-30

A simple, sensitive and selective spectrofluorimetric method for the determination of Metoclopramide hydrochloride (MCP) is developed. The MCP can remarkably enhances the luminescence intensity of the Eu(3+) ion doped in sol-gel matrix at lambda(ex)=380 nm in DMSO at pH 8.7. The intensity of the emission band of Eu(3+) ion doped in sol-gel matrix increases due to energy transfer from MCP to Eu(3+) in the excited state. The enhancement of the emission band of Eu(3+) ion doped in sol-gel matrix at 617 nm was found to be directly proportional to the concentration of MCP with a dynamic range of 5 x 10(-9) - 1.0 x 10(-6) mol L(-1) and detection limit of 2.2 x10(-11) mol L(-1). Copyright 2010 Elsevier B.V. All rights reserved.

Spectrofluorimetric analysis of famotidine in pharmaceutical preparations and biological fluids by derivatization with benzoin

NASA Astrophysics Data System (ADS)

Alamgir, Malik; Khuhawar, Muhammad Yar; Memon, Saima Q.; Hayat, Amir; Zounr, Rizwan Ali

2015-01-01

A sensitive and simple spectrofluorimetric method has been developed for the analysis of famotidine, from pharmaceutical preparations and biological fluids after derivatization with benzoin. The reaction was carried out in alkaline medium with measurement of fluorescence intensity at 446 nm with excitation wavelength at 286 nm. Linear calibration was obtained with 0.5-15 μg/ml with coefficient of determination (r2) 0.997. The factors affecting the fluorescence intensity were optimized. The pharmaceutical additives and amino acid did not interfere in the determination. The mean percentage recovery (n = 4) calculated by standard addition from pharmaceutical preparation was 94.8-98.2% with relative standard deviation (RSD) 1.56-3.34% and recovery from deproteinized spiked serum and urine of healthy volunteers was 98.6-98.9% and 98.0-98.4% with RSD 0.34-0.84% and 0.29-0.87% respectively.

Validated spectroflurimetric determination of some H1 receptor antagonist drugs in pharmaceutical preparations through charge transfer complexation.

PubMed

el-Din, Mohie K Sharaf; Ibrahim, Fawzia; Eid, Manal I; Wahba, Mary E K

2012-01-01

A validated simple, rapid, and selective spectrofluorimetric method was developed for the determination of some antihistaminic H(1) receptor antagonist drugs namely ebastine (EBS), cetirizine dihydrochloride (CTZ), and fexofenadine hydrochloride (FXD). The method is based on the reaction of the cited drugs with some Π acceptors namely p-chloranilic acid (CLA), tetracyanoethylene (TCNE), and 2,3-dichloro-5,6-dicyano-p-benzoquinone (DDQ) to give highly fluorescent derivatives. The fluorescence intensity-concentration plots were rectilinear over the concentration ranges of 0.2-3.0, 0.2-2.5 and 0.15-2.0 μg/ml for EBS with CLA, DDQ, and TCNE respectively; 0.5-7.0, 0.5-6.0, and 0.2-4.0 μg/ml for CTZ with the previously mentioned reagents, and 0.2-3.5, 0.5-6.0, and 0.2-3.5 μg/ml for FXD. The factors affecting the formation of the reaction products were carefully studied and optimized. The method was applied for the determination of the studied drugs in their dosage forms. The results obtained were in good agreement with those obtained by the comparison methods. Reactions Stoichiometries of the complexes formed between the studied drugs and Π acceptors were defined by the Job's method of the continuous variation and found in 1:1 in all cases.

Comparison between Two Linear Supervised Learning Machines' Methods with Principle Component Based Methods for the Spectrofluorimetric Determination of Agomelatine and Its Degradants.

PubMed

Elkhoudary, Mahmoud M; Naguib, Ibrahim A; Abdel Salam, Randa A; Hadad, Ghada M

2017-05-01

Four accurate, sensitive and reliable stability indicating chemometric methods were developed for the quantitative determination of Agomelatine (AGM) whether in pure form or in pharmaceutical formulations. Two supervised learning machines' methods; linear artificial neural networks (PC-linANN) preceded by principle component analysis and linear support vector regression (linSVR), were compared with two principle component based methods; principle component regression (PCR) as well as partial least squares (PLS) for the spectrofluorimetric determination of AGM and its degradants. The results showed the benefits behind using linear learning machines' methods and the inherent merits of their algorithms in handling overlapped noisy spectral data especially during the challenging determination of AGM alkaline and acidic degradants (DG1 and DG2). Relative mean squared error of prediction (RMSEP) for the proposed models in the determination of AGM were 1.68, 1.72, 0.68 and 0.22 for PCR, PLS, SVR and PC-linANN; respectively. The results showed the superiority of supervised learning machines' methods over principle component based methods. Besides, the results suggested that linANN is the method of choice for determination of components in low amounts with similar overlapped spectra and narrow linearity range. Comparison between the proposed chemometric models and a reported HPLC method revealed the comparable performance and quantification power of the proposed models.

Spectrofluorimetric study on the inclusion interaction between vitamin K 3 with p-( p-sulfonated benzeneazo)calix[6]arene and determination of VK 3

NASA Astrophysics Data System (ADS)

Zhou, Yunyou; Xu, Hongwei; Wu, Lian; Liu, Chun; Lu, Qin; Wang, Lun

2008-11-01

The characteristics of host-guest complexation between p-( p-sulfonated benzeneazo) calix[6]arene (SBC6A) and vitamin K 3 (VK 3) were investigated by fluorescence spectrometry. A 1:1 stoichiometry for the complexation was established and was verified by Job's plot. An association constant of 4.95 × 10 3 L mol -1 at 20 °C was calculated by applying a deduced equation. The interaction mechanism of the inclusion complex was discussed. It was found that the fluorescence of SBC6A could be remarkably quenched by an appropriate amount of VK 3 especially when non-ionic surfactant Triton X-100 existed. According to the obtained results, a novel sensitive spectrofluorimetric method for the determination of VK 3 based on supramolecular complex was developed with a linear range of 5.0 × 10 -7-3.0 × 10 -5 mol L -1 and a detection limit of 2.0 × 10 -7 mol L -1. The proposed method was used to determine VK 3 in commercial preparations with satisfactory results.

Spectrofluorimetric study on the inclusion interaction between vitamin K3 with p-(p-sulfonated benzeneazo)calix[6]arene and determination of VK3.

PubMed

Zhou, Yunyou; Xu, Hongwei; Wu, Lian; Liu, Chun; Lu, Qin; Wang, Lun

2008-11-15

The characteristics of host-guest complexation between p-(p-sulfonated benzeneazo) calix[6]arene (SBC6A) and vitamin K3 (VK3) were investigated by fluorescence spectrometry. A 1:1 stoichiometry for the complexation was established and was verified by Job's plot. An association constant of 4.95 x 10(3)L mol(-1) at 20 degrees C was calculated by applying a deduced equation. The interaction mechanism of the inclusion complex was discussed. It was found that the fluorescence of SBC6A could be remarkably quenched by an appropriate amount of VK3 especially when non-ionic surfactant Triton X-100 existed. According to the obtained results, a novel sensitive spectrofluorimetric method for the determination of VK3 based on supramolecular complex was developed with a linear range of 5.0 x 10(-7) -3.0 x 10(-5)mol L(-1) and a detection limit of 2.0 x 10(-7)mol L(-1). The proposed method was used to determine VK3 in commercial preparations with satisfactory results.

Determination of Organothiophosphate Insecticides in Environmental Water Samples by a Very Simple and Sensitive Spectrofluorimetric Method.

PubMed

Bavili Tabrizi, Ahad; Abdollahi, Ali

2015-10-01

A simple, rapid and sensitive spectrofluorimetric method was developed for the determination of di-syston, ethion and phorate in environmental water samples. The procedure is based on the oxidation of these pesticides with cerium (IV) to produce cerium (III), and its fluorescence was monitored at 368 ± 3 nm after excitation at 257 ± 3 nm. The variables effecting oxidation of each pesticide were studied and optimized. Under the experimental conditions used, the calibration graphs were linear over the range 0.2-15, 0.1-13, 0.1-13 ng mL(-1) for di-syston, ethion and phorate, respectively. The limit of detection and quantification were in the range 0.034-0.096 and 0.112-0.316 ng mL(-1), respectively. Intra- and inter-day assay precisions, expressed as the relative standard deviation (RSD), were lower than 5.2 % and 6.7 %, respectively. Good recoveries in the range 86 %-108 % were obtained for spiked water samples. The proposed method was applied to the determination of studied pesticides in environmental water samples.

Development and validation spectroscopic methods for the determination of lomefloxacin in bulk and pharmaceutical formulations

NASA Astrophysics Data System (ADS)

El-Didamony, A. M.; Hafeez, S. M.

2016-01-01

Four simple, sensitive spectrophotometric and spectrofluorimetric methods (A-D) for the determination of antibacterial drug lomefloxacin (LMFX) in pharmaceutical formulations have been developed. Method A is based on formation of ternary complex between Pd(II), eosin and LMFX in the presence of methyl cellulose as surfactant and acetate-HCl buffer pH 4.0. Spectrophotometrically, under the optimum conditions, the ternary complex showed absorption maximum at 530 nm. Methods B and C are based on redox reaction between LMFX and KMnO4 in acid and alkaline media. In indirect spectrophotometry method B the drug solution is treated with a known excess of KMnO4 in H2SO4 medium and subsequent determination of unreacted oxidant by reacting it with safronine O in the same medium at λmax = 520 nm. Direct spectrophotometry method C involves treating the alkaline solution of LMFX with KMnO4 and measuring the bluish green product at 604 nm. Method D is based on the chelation of LMFX with Zr(IV) to produce fluorescent chelate. At the optimum reaction conditions, the drug-metal chelate showed excitation maxima at 280 nm and emission maxima at 443 nm. The optimum experimental parameters for the reactions have been studied. The validity of the described procedures was assessed. Statistical analysis of the results has been carried out revealing high accuracy and good precision. The proposed methods were successfully applied for the determination of the selected drug in pharmaceutical preparations with good recoveries.

Stability-indicating spectrofluorimetric method for determination of itopride hydrochloride in raw material and pharmaceutical formulations.

PubMed

Walash, Mohamed I; Ibrahim, Fawzia; Eid, Manal I; El Abass, Samah Abo

2013-11-01

A simple, sensitive and rapid spectrofluorimetric method for determination of itopride hydrochloride in raw material and tablets has been developed. The proposed method is based on the measurement of the native fluorescence of the drug in water at 363 nm after excitation at 255 nm. The relative fluorescence intensity-concentration plot was rectilinear over the range of 0.1-2 μg/mL (2.5 × 10(-7)-5.06 × 10(-6) mole/L), with good correlation (r = 0.9999), limit of detection of 0.015 μg/mL and a lower limit of quantification of 0.045 μg/mL. The described method was successfully applied for the determination of itopride hydrochloride in its commercial tablets with average percentage recovery of 100.11 ± 0.32 without interference from common excipients. Additionally, the proposed method can be applied for determination of itopride in combined tablets with rabeprazole or pantoprazole without prior separation. The method was extended to stability study of itopride. The drug was exposed to acidic, alkaline, oxidative and photolytic degradation according to ICH guidelines. Moreover, the method was utilized to investigate the kinetics of the alkaline, acidic and oxidative degradation of the drug. A proposal for the degradation pathways was postulated.

Aluminium sensitized spectrofluorimetric determination of fluoroquinolones in milk samples coupled with salting-out assisted liquid-liquid ultrasonic extraction

NASA Astrophysics Data System (ADS)

Xia, Qinghai; Yang, Yaling; Liu, Mousheng

2012-10-01

An aluminium sensitized spectrofluorimetric method coupled with salting-out assisted liquid-liquid ultrasonic extraction for the determination of four widely used fluoroquinolones (FQs) namely norfloxacin (NOR), ofloxacin (OFL), ciprofloxacin (CIP) and gatifloxacin (GAT) in bovine raw milk was described. The analytical procedure involves the fluorescence sensitization of aluminium (Al3+) by complexation with FQs, salting-out assisted liquid-liquid ultrasonic extraction (SALLUE), followed by spectrofluorometry. The influence of several parameters on the extraction (the salt species, the amount of salt, pH, temperature and phase volume ratio) was investigated. Under optimized experimental conditions, the detection limits of the method in milk varied from 0.009 μg/mL for NOR to 0.016 μg/mL for GAT (signal-to-noise ratio (S/N) = 3). The relative standard deviations (RSD) values were found to be relatively low (0.54-2.48% for four compounds). The calibration graph was linear from 0.015 to 2.25 μg/mL with coefficient of determinations not less than 0.9974. The methodology developed was applied to the determination of FQs in bovine raw milk samples. The main advantage of this method is simple, accurate and green. The method showed promising applications for analyzing polar analytes especially polar drugs in various sample matrices.

Partial least-squares with residual bilinearization for the spectrofluorimetric determination of pesticides. A solution of the problems of inner-filter effects and matrix interferents.

PubMed

Piccirilli, Gisela N; Escandar, Graciela M

2006-09-01

This paper demonstrates for the first time the power of a chemometric second-order algorithm for predicting, in a simple way and using spectrofluorimetric data, the concentration of analytes in the presence of both the inner-filter effect and unsuspected species. The simultaneous determination of the systemic fungicides carbendazim and thiabendazole was achieved and employed for the discussion of the scopes of the applied second-order chemometric tools: parallel factor analysis (PARAFAC) and partial least-squares with residual bilinearization (PLS/RBL). The chemometric study was performed using fluorescence excitation-emission matrices obtained after the extraction of the analytes over a C18-membrane surface. The ability of PLS/RBL to recognize and overcome the significant changes produced by thiabendazole in both the excitation and emission spectra of carbendazim is demonstrated. The high performance of the selected PLS/RBL method was established with the determination of both pesticides in artificial and real samples.

Spectrofluorimetric analysis of famotidine in pharmaceutical preparations and biological fluids by derivatization with benzoin.

PubMed

Alamgir, Malik; Khuhawar, Muhammad Yar; Memon, Saima Q; Hayat, Amir; Zounr, Rizwan Ali

2015-01-05

A sensitive and simple spectrofluorimetric method has been developed for the analysis of famotidine, from pharmaceutical preparations and biological fluids after derivatization with benzoin. The reaction was carried out in alkaline medium with measurement of fluorescence intensity at 446 nm with excitation wavelength at 286 nm. Linear calibration was obtained with 0.5-15 μg/ml with coefficient of determination (r(2)) 0.997. The factors affecting the fluorescence intensity were optimized. The pharmaceutical additives and amino acid did not interfere in the determination. The mean percentage recovery (n=4) calculated by standard addition from pharmaceutical preparation was 94.8-98.2% with relative standard deviation (RSD) 1.56-3.34% and recovery from deproteinized spiked serum and urine of healthy volunteers was 98.6-98.9% and 98.0-98.4% with RSD 0.34-0.84% and 0.29-0.87% respectively. Copyright © 2014 Elsevier B.V. All rights reserved.

Utility of Von Pechman synthesis of coumarin reaction for development of spectrofluorimetric method for quantitation of salmeterol xinafoate in pharmaceutical preparations and human plasma.

PubMed

Awad, Mohamed; Hammad, Mohamed A; Abdel-Megied, Ahmed M; Omar, Mahmoud A

2018-04-30

Simple, precise and selective spectrofluorimetric technique was evolved for quantitation of selective β 2 agonist drug namely salmeterol xinafoate (SAL). Utilizing its phenolic nature, a method was described based on the reaction of the studied drug with ethyl acetoacetate (EAA) to yield extremely fluorescent coumarin product which can be detected at 480 nm (λ ex  = 420 nm). The procedure obeys Beer's law with a correlation coefficient of r = 0.9999 in the concentration range between 500 and 5000 ng ml -1 with and 177 ng ml -1 for limit of detection (LOD) and limit of quantification (LOQ), respectively. Diverse reaction variables influencing the firmness and formation of the coumarin product were accurately examined and modified to ensure greatest sensitivity of the procedure. The proposed technique was performed and examined according to the US Food and Drug Administration (FDA) guidelines for bio-analytical methods and was efficiently applied for quantitation of SAL in both pharmaceutical preparations (% recovery = 100.06 ± 1.07) and spiked human plasma (% recovery = 96.64-97.14 ± 1.01-1.52). Copyright © 2018 John Wiley & Sons, Ltd.

Micelle-enhanced and terbium-sensitized spectrofluorimetric determination of gatifloxacin and its interaction mechanism

NASA Astrophysics Data System (ADS)

Guo, Changchuan; Wang, Lei; Hou, Zhun; Jiang, Wei; Sang, Lihong

2009-05-01

A terbium-sensitized spectrofluorimetric method using an anionic surfactant, sodium dodecyl benzene sulfonate (SDBS), was developed for the determination of gatifloxacin (GFLX). A coordination complex system of GFLX-Tb 3+-SDBS was studied. It was found that SDBS significantly enhanced the fluorescence intensity of the complex (about 11-fold). Optimal experimental conditions were determined as follows: excitation and emission wavelengths of 331 and 547 nm, pH 7.0, 2.0 × 10 -4 mol l -1 terbium (III), and 2.0 × 10 -4 mol l -1 SDBS. The enhanced fluorescence intensity of the system (Δ If) showed a good linear relationship with the concentration of GFLX over the range of 5.0 × 10 -10 to 5.0 × 10 -8 mol l -1 with a correlation coefficient of 0.9996. The detection limit (3 σ) was determined as 6.0 × 10 -11 mol l -1. This method has been successfully applied to the determination of GFLX in pharmaceuticals and human urine/serum samples. Compared with most of other methods reported, the rapid and simple procedure proposed in the text offers higher sensitivity, wider linear range, and better stability. The interaction mechanism of the system is also studied by the research of ultraviolet absorption spectra, surface tension, solution polarity and fluorescence polarization.

Spectrofluorimetric determination of thallium in silicate rocks with rhodamine b in the presence of aluminum chloride

USGS Publications Warehouse

Shnepfe, M.M.

1975-01-01

A sensitive spectrofluorimetric procedure with rhodamine B in the presence of aluminum chloride is given for determining submicrogram and microgram quantities of thallium in silicate rocks. Samples are decomposed with a mixture of hydrofluoric and nitric acids and then treated with hydrochloric acid. Thallium is extracted as its dithizonate with chloroform from an alkaline medium containing ascorbate, citrate, and cyanide and then back-extracted with dilute nitric acid. After destruction of the organic matter and treatment with bromine, hydrochloric acid, aluminum chloride, and rhodamine B, the {A table is presented}. ?? 1975.

Ion Sensor Properties of Fluorescent Schiff Bases Carrying Dipicolylamine Groups. A Simple Spectrofluorimetric Method to Determine Cu (II) in Water Samples.

PubMed

Vanlı, Elvan; Mısır, Miraç Nedim; Alp, Hakan; Ak, Tuğba; Özbek, Nurhayat; Ocak, Ümmühan; Ocak, Miraç

2017-09-01

Four fluorescent Schiff bases carrying dipicolylamine groups were designed and synthesized to determine their ion sensor properties in partial aqueous solution. The corresponding amine compound and the aldehyde compounds such as 1-naphthaldehyde, 9-anthraldehyde, phenanthrene-9-carboxaldehyde and 1-pyrenecarboxaldehyde were used to prepare the new Schiff bases. The influence of many metal cations and anions on the spectroscopic properties of the ligands was investigated in ethanol-water (1:1) by means of emission spectrometry. From the spectrofluorimetric titrations, the complexation stoichiometry and complex stability constants of the ligands with Cd 2+ , Zn 2+ , Cu 2+ and Hg 2+ ions were determined. The ligands did not interact with the anions. However, the Schiff base derived from phenanthrene-9-carboxaldehyde showed sensitivity for Cu 2+ among the tested metal ions. The phenanthrene-based Schiff base was used as analytical ligand for the simple and fast determination of Cu 2+ ion in water samples. A modified standard addition method was used to eliminate matrix effect. The linear range was from 0.3 mg/L to 3.8 μg/L. Detection and quantification limits were 0.14 and 0.43 mg/L, respectively. Maximum contaminant level goal (MCLG) for copper in drinking water according to EPA is 1.3 mg/L. The proposed method has high sensitivity to determine copper in drinking waters.

An alternative derivatization reaction to the determination of doramectin in bovine milk using spectrofluorimetry

NASA Astrophysics Data System (ADS)

Maia, Priscila M. S.; de F. Rezende, Flavia B.; Netto, Annibal D. Pereira; de C. Marques, Flávia F.

The doramectin (DOR), which belongs to the avermectins group (AVM), has a high antiparasitic activity and so it has been widely used in food-producing animals. The DOR shows low fluorescence quantum efficiency and as a consequence, chemical derivatization reactions are necessary to produce derivatives with improved luminescent properties before its determination by fluorimetry. As the presence of this compound in food represents a risk to human health, an easy, clean and low cost derivatization reaction, which is alternative to those usually employed and that enables its spectrofluorimetric determination in milk samples, was developed. Ethanolic solutions of DOR, containing sodium hydroxide at a final concentration of 0.25 mol L-1, after 60 min of heating at 50 °C, produced fluorescent signals 1000 times higher than the original ethanolic solution. Using these optimized conditions, a linear response range that extended from 50.00 to 1000 μg L-1, with a value of (R2) equal to 0.9970, was obtained. Average recovery of DOR was 92.5 ± 1.5% (n = 3) in bovine milk fortified samples submitted to a liquid-liquid extraction at low temperature and pre concentration process, indicating the usefulness and effectiveness of the proposed method. The proposed spectrofluorimetric method is an alternative to high-performance liquid chromatography (HPLC) based methods, allowing rapid and simple detection of doramectin in milk samples.

Simultaneous determination of hydroquinone, resorcinol, phenol, m-cresol and p-cresol in untreated air samples using spectrofluorimetry and a custom multiple linear regression-successive projection algorithm.

PubMed

Pistonesi, Marcelo F; Di Nezio, María S; Centurión, María E; Lista, Adriana G; Fragoso, Wallace D; Pontes, Márcio J C; Araújo, Mário C U; Band, Beatriz S Fernández

2010-12-15

In this study, a novel, simple, and efficient spectrofluorimetric method to determine directly and simultaneously five phenolic compounds (hydroquinone, resorcinol, phenol, m-cresol and p-cresol) in air samples is presented. For this purpose, variable selection by the successive projections algorithm (SPA) is used in order to obtain simple multiple linear regression (MLR) models based on a small subset of wavelengths. For comparison, partial least square (PLS) regression is also employed in full-spectrum. The concentrations of the calibration matrix ranged from 0.02 to 0.2 mg L(-1) for hydroquinone, from 0.05 to 0.6 mg L(-1) for resorcinol, and from 0.05 to 0.4 mg L(-1) for phenol, m-cresol and p-cresol; incidentally, such ranges are in accordance with the Argentinean environmental legislation. To verify the accuracy of the proposed method a recovery study on real air samples of smoking environment was carried out with satisfactory results (94-104%). The advantage of the proposed method is that it requires only spectrofluorimetric measurements of samples and chemometric modeling for simultaneous determination of five phenols. With it, air is simply sampled and no pre-treatment sample is needed (i.e., separation steps and derivatization reagents are avoided) that means a great saving of time. Copyright © 2010 Elsevier B.V. All rights reserved.

Two different spectrofluorimetric methods for simultaneous determination of gemfibrozil and rosiglitazone in human plasma.

PubMed

El-Din, Mohie M K Sharaf; Attia, Khalid A M; Nassar, Mohamed W I; Kaddah, Mohamed M Y

2010-10-15

Two accurate, reliable, and highly sensitive spectrofluorimetric methods were developed for simultaneous determination of binary mixture gemfibrozil and rosiglitazone in human plasma without prior separation steps. The first method is based on synchronous fluorescence spectrometry using double scans. At Δλ=27nm, gemfibrozil yields detectable signal that is independent of the presence of rosiglitazone. Similarly, at Δλ=120nm the signal of rosiglitazone is not influenced by the presence of gemfibrozil. Signals at two wavelengths, 301 (Δλ=27nm) and 368nm (Δλ=120nm) vary linearly with gemfibrozil and rosiglitazone concentrations over the range 100-700ngmL(-1) (for gemfibrozil) and 20-140ngmL(-1) (for rosiglitazone), respectively. The limits of detection (LOD) were 2.3 and 2.72ngmL(-1) for gemfibrozil and rosiglitazone, respectively. The second method is based on the technique of simultaneous equations (Vierodt's method), in which 258nm was selected as the excitation wavelength. Two equations are constructed based on the fact that at ( λ(EM)₂=302 nm of gemfibrozil) and (λ(EM)₂=369 nm of rosiglitazone) the fluorescence of the mixture is the sum of the individual fluorescence of gemfibrozil and rosiglitazone. The limits of detection (LOD) were 28.1 and 23.63ngmL(-1) for gemfibrozil and rosiglitazone, respectively. The proposed methods were successfully applied for the determination of the two compounds in synthetic mixtures and in human plasma with a good recovery. Copyright © 2010 Elsevier B.V. All rights reserved.

Highly sensitive spectrofluorimetric determination of lomefloxacin in spiked human plasma, urine and pharmaceutical preparations.

PubMed

Ulu, Sevgi Tatar

2009-09-01

A sensitive, simple and selective spectrofluorimetric method was developed for the determination of lomefloxacin in biological fluids and pharmaceutical preparations. The method is based on the reaction between the drug and 4-chloro-7-nitrobenzodioxazole in borate buffer of pH 8.5 to yield a highly fluorescent derivative that is measured at 533 nm after excitation at 433 nm. The calibration curves were linear over the concentration ranges of 12.5-625, 15-1500 and 20-2000 ng/mL for plasma, urine and standard solution, respectively. The limits of detection were 4.0 ng/mL in plasma, 5.0 ng/mL in urine and 7.0 ng/mL in standard solution. The intra-assay accuracy and precision in plasma ranged from 0.032 to 2.40% and 0.23 to 0.36%, respectively, while inter-assay accuracy and precision ranged from 0.45 to 2.10% and 0.25 to 0.38%, respectively. The intra-assay accuracy and precision estimated on spiked samples in urine ranged from 1.27 to 4.20% and 0.12 to 0.24%, respectively, while inter-assay accuracy and precision ranged from 1.60 to 4.00% and 0.14 to 0.25%, respectively. The mean recovery of lomefloxacin from plasma and urine was 98.34 and 98.43%, respectively. The method was successfully applied to the determination of lomefloxacin in pharmaceuticals and biological fluids.

Spectrofluorimetric assessment of chlorzoxazone and ibuprofen in pharmaceutical formulations by using Eu-tetracycline HCl optical sensor doped in sol-gel matrix.

PubMed

Attia, M S; Ramsis, M N; Khalil, L H; Hashem, S G

2012-03-01

A novel, simple, sensitive and selective spectrofluorimetric method was developed for the determination of trace amounts of chlorzoxazone and Ibuprofen in pharmaceutical tablets using optical sensor Eu-Tetracycline HCl doped in sol-gel matrix. The chlorzoxazone or Ibuprofen can remarkably enhance the luminescence intensity of Eu-Tetracycline HCl complex doped in a sol-gel matrix in dimethylformamide (DMF) at pH 9.7 and 6.3, respectively, λ(ex) = 400 nm. The enhancing of luminescence intensity peak of Eu-Tetracycline HCl complex at 617 nm is proportional to the concentration of chlorzoxazone or Ibuprofen a result that suggested profitable application as a simple optical sensor for chlorzoxazone or Ibuprofen assessment. The dynamic ranges found for the determination of chlorzoxazone and Ibuprofen concentration are 5 × 10(-9)-1 × 10(-4) and 1 × 10(-8)-7 × 10(-5) mol L(-1), and the limit of detection (LOD) and quantitation limit of detection (LOQ) are 3.1 × 10(-10), 9.6 × 10(-10) and 5.6 × 10(-10), 1.7 × 10(-9) mol L(-1), respectively.

Detection and quantification of the selective EP4 receptor antagonist CJ-023423 (grapiprant) in canine plasma by HPLC with spectrofluorimetric detection.

PubMed

Vito, Virgina De; Saba, Alessandro; Lee, Hong-Ki; Owen, Helen; Poapolathep, Amnart; Giorgi, Mario

2016-01-25

Grapiprant, a novel pharmacologically active ingredient, acts as a selective EP4 receptor antagonist whose physiological ligand is prostaglandin E2 (PGE2). It is currently under development for use in humans and dogs for the control of pain and inflammation associated with osteoarthritis. The aim of the present study was to develop an easy and sensitive method to quantify grapiprant in canine plasma and to apply the method in a canine patient. Several parameters, both in the extraction and detection method were evaluated. The final mobile phase consisted of ACN:AcONH4 (20 mM) solution, pH 4 (70:30, v/v) at a flow rate of 1 mL/min. The elution of grapiprant and IS (metoclopramide) was carried out in isocratic mode through a Synergi Polar-RP 80A analytical column (150 mm × 4.6 mm). The best excitation and emission wavelengths were 320 and 365 nm, respectively. Grapiprant was extracted from the plasma using CHCl3, which gave a recovery of 88.1 ± 10.22% and a lower limit of quantification (LLOQ) of 10 ng/mL. The method was validated in terms of linearity, limit of detection (LOD), LLOQ, selectivity, accuracy and precision, extraction recovery, stability, and inter-laboratory cross validation, according to international guidelines. The chromatographic runs were specific with no interfering peaks at the retention times of the analyte and IS, as confirmed by HPLC-MS experiments. In conclusion, this was a simple and effective method using HPLC-FL to detect grapiprant in plasma, which may be useful for future pharmacokinetic studies. Copyright © 2015 Elsevier B.V. All rights reserved.

An eco-friendly stability-indicating spectrofluorimetric method for the determination of two anticancer stereoisomer drugs in their pharmaceutical preparations following micellar enhancement: Application to kinetic degradation studies.

PubMed

El-Kimary, Eman I; El-Yazbi, Amira F

2016-06-15

A new rapid and highly sensitive stability-indicating spectrofluorimetric method was developed for the determination of two stereoisomers anticancer drugs, doxorubicin (DOX) and epirubicin (EPI) in pure form and in pharmaceutical preparations. The fluorescence spectral behavior of DOX and EPI in a sodium dodecyl sulfate (SDS) micellar system was investigated. It was found that the fluorescence intensity of DOX and EPI in an aqueous solution of phosphate buffer pH4.0 and in the presence of SDS was greatly (about two fold) enhanced and the mechanism of fluorescence enhancement effect of SDS on DOX was also investigated. The fluorescence intensity of DOX or EPI was measured at 553nm after excitation at 497nm. The plots of fluorescence intensity versus concentration were rectilinear over a range of 0.03-2μg/mL for both DOX and EPI with good correlation coefficient (r>0.999). High sensitivity to DOX and EPI was attained using the proposed method with limits of detection of 10 and 9ng/mL and limits of quantitation of 29 and 28ng/mL, for DOX and EPI, respectively. The method was successfully applied for the determination of DOX and EPI in biological fluids and in their commercial pharmaceutical preparations and the results were concordant with those obtained using a previously reported method. The application of the proposed method was extended to stability studies of DOX following different forced degradation conditions (acidic, alkaline, oxidative and photolytic) according to ICH guidelines. Moreover, the kinetics of the alkaline and oxidative degradation of DOX was investigated and the apparent first-order rate constants and half-life times were calculated. Copyright © 2016 Elsevier B.V. All rights reserved.

Plackett-Burman and Box-Behnken designs as chemometric tools for micro-determination of L-Ornithine

NASA Astrophysics Data System (ADS)

Elazazy, Marwa S.; El-Hamshary, Marwa; Sakr, Marwa; Al-Easa, Hala S.

2018-03-01

Plackett-Burman (PB) and Box-Behnken (BB) screening and response surface factorial designs were used to evaluate spectrophotometric and spectrofluorimetric approaches for the determination of L-Ornithine (ORN) as per se and in dietary supplements. Both approaches were based on the derivatization of the primary amino group of ORN via Hantzsch condensation reaction producing yellow coloured adducts (dihydrolutidine derivative). The reaction product was determined spectrophotometrically (method A) at λmax = 327 nm and spectrofluorimetrically (method B) at 480 nm (λem) after excitation at 325 nm (λex). A multivariate scheme was tailored to investigate the process numerical variables; reaction temperature, heating time, reagent volume, and pH implementing PB as a screening design followed by BB as an optimization strategy. Categorical factors including diluting solvent and sequence of addition were kept invariable. Responses of the reaction systems were the maximum absorbance (Y1) and maximum fluorescence intensity (Y2), correspondingly. Quality tools as well as ANOVA testing, before and after response transformation were used to decide upon the substantial variables. Following the optimization of reaction variables using desirability plots, calibration graphs were found to be rectilinear in the range of 6-14 μg/mL and 0.4-1.2 μg/mL for methods A and B, respectively. Both methods proved to be sensitive with detection limits (DL) of 337 and 85 ng/mL, and quantitation limits (QL) of 1086 and 283 ng/mL, for methods A and B, respectively. An interference study was performed using potential foreign species. No significant interference effect was observed on any of the proposed procedures. System performance was addressed following ICH guidelines and considering parameters such as linearity, detection and quantification limits, accuracy and precision, robustness and specificity.

Three-way spectrofluorimetric-assisted multivariate determination of nonylphenol ethoxylate and 2-naphtalene sulfonate in wastewater samples and optimization approach for their photocatalytic degradation by CoTiO3 nanostructure.

PubMed

Mohsenikia, Atefeh; Gholami, Ali; Masoum, Saeed; Abbasi, Saleheh

2017-09-01

This study presents a new strategy for the simultaneous quantification of two industrial contaminants. The excitation-emission fluorescence data matrix combined with a three-way chemometric method, such as parallel factor analysis, was used for the determination of nonylphenol ethoxylate (NPE-9) as a nonionic surfactant and 2-naphthalene sulfonate (2-NS) in waste water samples. It is noticeable that this method can resolve overlapping signal into spectral and relative concentration profiles. By spiking the known concentrations of these compounds in the wastewater samples, the accuracy of the proposed methods was validated and recoveries of the spiked values were calculated. High recoveries (i.e. 90-110%) obtained for the waste water samples indicate the present method can be used successfully to determine the analytes concentration in the environmental contaminations. The photocatalytic degradation of NPE-9 and 2-NS in aqueous solution was studied using the CoTiO 3 nanoparticles catalyst. It was synthesized by the sol-gel technique. The catalytic activity of the prepared nanoparticles was measured in a batch photoreactor containing appropriate solutions of these compounds with UV irradiation. The photodegradation process of these compounds was optimized by using the central composite design. The CoTiO 3 showed high activity for UV-photocatalytic degradation of NPE-9 and 2-NS.

Surfactant-enhanced spectrofluorimetric determination of total aflatoxins from wheat samples after magnetic solid-phase extraction using modified Fe3O4 nanoparticles

NASA Astrophysics Data System (ADS)

Manafi, Mohammad Hanif; Allahyari, Mehdi; Pourghazi, Kamyar; Amoli-Diva, Mitra; Taherimaslak, Zohreh

2015-07-01

The extraction and preconcentration of total aflatoxins (including aflatoxin B1, B2, G1, and G2) using magnetic nanoparticles based solid phase extraction (MSPE) followed by surfactant-enhanced spectrofluorimetric detection was proposed. Ethylene glycol bis-mercaptoacetate modified silica coated Fe3O4 nanoparticles as an efficient antibody-free adsorbent was successfully applied to extract aflatoxins from wheat samples. High surface area and strong magnetization properties of magnetic nanoparticles were utilized to achieve high enrichment factor (97), and satisfactory recoveries (92-105%) using only 100 mg of the adsorbent. Furthermore, the fast separation time (less than 10 min) avoids many time-consuming cartridge loading or column-passing procedures accompany with the conventional SPE. In determination step, signal enhancement was performed by formation of Triton X-100 micelles around the analytes in 15% (v/v) acetonitrile-water which dramatically increase the sensitivity of the method. Main factors affecting the extraction efficiency and signal enhancement of the analytes including pH of sample solution, desorption conditions, extraction time, sample volume, adsorbent amount, surfactant concentration and volume and time of micelle formation were evaluated and optimized. Under the optimum conditions, wide linear range of 0.1-50 ng mL-1 with low detection limit of 0.03 ng mL-1 were obtained. The developed method was successfully applied to the extraction and preconcentration of aflatoxins in three commercially available wheat samples and the results were compared with the official AOAC method.

Spectrofluorimetric determination of cefixime using terbium-danofloxacin probe

PubMed Central

Manzoori, Jamshid L.; Amjadi, Mohammad; Soltani, Naser; Jouyban, Abolghasem

2014-01-01

Objective(s): Cefixime (Cfx), is a semi-synthetic third-generation oral cephalosporin antibiotic that is prescribed for the treatment of susceptible infections. There are some procedures for the determination of Cfx in pharmaceutical formulations and biological samples. Herein a spectrofluorimetric method was proposed for Cfx determination based on the fluorescence quenching of terbium-danofloxacin (Tb3+-Dano) in the presence of Cfx. Materials and Methods: Cfx was detected based on fluorescence quenching of terbium-danofloxacin (Tb3+-Dano) in the presence of Cfx with maximum excitation and emission wavelengths at 347 nm and 545 nm, respectively. The quenched fluorescence intensity of Tb3+- Dano system is proportional to the concentration of Cfx. The optimum conditions for the determination of Cfx were studied. Results: The maximum response was achieved under optimum conditions of [Tris buffer]= 0.008 mol/l (pH 6.5), [Tb3+]=1×10-4 mol/l and [Dano]=1×10-4 mol/l. The developed method was evaluated in terms of accuracy, precision and limit of detection. The linear concentration ranges for quantification of Cfx were 8.8×10-8-8.8×10-7 mol/l and 1.1×10-7-8.8×10-7 mol/l in standard and human serum samples with the detection limits (S/N=3) of 2.8×10-8 mol/l and 3.9×10-8 mol/l, respectively. The Cfx was determined in pharmaceutical tablets and spiked serum samples and the results were satisfactory. Conclusion: This method is simple, practical and relatively interference-free for determination of Cfx in pharmaceutical tablets and serum samples. PMID:24904718

Spectrofluorimetric determination of hydrogen peroxide with 2-hydroxy-1-naphthaldehyde salicyloylhydrazone (HNSH) as the substrate for horseradish peroxidase (HRP).

PubMed

Tang, Bo; Wang, Yan; Sun, Ying; Shen, Han Xi

2002-01-01

The oxidation reaction of HNSH with H2O2 under the catalysis of HRP was studied in detail. The possible reaction mechanism was discussed. Under optimum experimental conditions, the oxidized product of HNSH had excitation and emission maxima at 296 and 414 nm, respectively. A study to prove the existence of -O-O-H in polyethylene glycols was carried out. The proposed method was successfully applied to the determination of -O-O-H in polyethylene glycols.

Spectrofluorimetric determination of 1,3,5-trinitro-1,3,5-triazacyclohexane (hexogen, RDX) as a nitramine type explosive.

PubMed

Lapat, A; Székelyhidi, L; Hornyák, I

1997-01-01

RDX is one of the most important military explosives. It is a component of some plastic explosives which are frequently used in terrorist attacks. Two fluorimetric methods have been described for the quantitative determination of RDX which are based on the detection of nitrite ions. After a basic decomposition of RDX the nitrite ion can be detected by reaction with 4-aminofluorescein and by reactions forming a lumogallion-gallium(III) complex. These fluorimetric methods have been compared to a photometric reaction (Griess reaction) for determination of nitrite ions. It has been found that the fluorimetric methods have a higher sensitivity than the photometric method and they have been used in a wider concentration range.

Enzymatic activity in the surface microlayer and subsurface water in the harbour channel

NASA Astrophysics Data System (ADS)

Perliński, Piotr; Mudryk, Zbigniew J.; Antonowicz, Józef

2017-09-01

Hydrolytic activity of eight extracellular enzymes was determined spectrofluorimetric method in the surface microlayer and subsurface water in the harbour channel in Ustka. The ranking order of the potential enzyme activity rates in the studied water layers was as follows: lipase > phosphatase > aminopeptidase > β-glucosidase > α-glucosidase > xylanase > cellulase > chitinase. The level of activity of all studied hydrolases was higher in the surface microlayer than subsurface water. No clear gradients in the level of enzymatic activity were determined along the horizontal profile of the studied channel. Activity of extracellular enzymes was strongly influenced by the season.

Determination of human serum albumin using an intramolecular charge transfer fluorescence probe: 4'-dimethylamino-2,5-dihydroxychalcone.

PubMed

Xu, Zhicheng; Yang, Weibing; Dong, Chuan

2005-09-15

A new intramolecular charge transfer fluorescence probe, namely, 4'-dimethylamino-2,5-dihydroxychalcone (DMADHC), exhibited dramatic enhancement of fluorescence intensity with an accompanying blue shift of the emission maximum when the concentration of human serum albumin (HSA) was increased. Binding to HSA also caused a progressive shift in the absorption spectrum of DMADHC, and a clear isosbestic point appeared. The binding site number and binding constant were calculated. Thermodynamic parameters were given and possible binding site was speculated. The optimum conditions for the determination of HSA were also investigated. A new, fast, and simple spectrofluorimetric method for the determination of HSA was developed. In the detection of HSA in samples of human plasma, this method gave values close to that of the Erythrosin B method.

Spectrofluorimetric study of the interaction of the mycotoxin citrinin with gold nanoparticles

USDA-ARS?s Scientific Manuscript database

Citrinin is a nephrotoxic secondary metabolite produced by certain fungal species from the Aspergillus, Penicillium, and Monascus genera. Citrinin producing species occasionally contaminate grains, and may pose food safety risks. Analysis of this mycotoxin is frequently carried out by liquid chromat...

An evaluation of the automated assay of urinary oestrogens in pregnant women

PubMed Central

Muir, G. G.; Ryan, M.; Conaill, D. U.

1970-01-01

An automated assay suitable for estimating urinary oestrogens in pregnant women has been investigated. Fluorimetry was found to have considerable advantages over colorimetry. The fluorimetric assay was simpler, more precise, more sensitive, and eliminated the need for correction for non-specific chromogens; in the assay of oestriol in pregnant women there was no need for correction for non-specific fluorescence. Spectrofluorimetric and photometric analyses, recoveries, and reproducibility show that the method offers a robust means of providing values for urinary oestrogen in pregnant women on a scale of up to 100 tests a day, the time of the assay being one and a half hours. PMID:5476876

[Determination of iron(III) in Chinese herbal medicine and tea based on fluorescence quenching of 2,4-dichro-phenylfluorone].

PubMed

Dai, Gang; Aodeng, Gao-wa

2004-07-01

A new spectrofluorimetric method for the determination of trace amount of Fe(III) has been developed. This method is based on the fluorescence quenching of 2,4-dichro-phenylfluorone due to the formation of complex Fe(III)-DCIPF. In pH 5.2-5.9 buffer solution, Fe (III) and DCIPF react on each other to form a red complex. Its composition was established by method of continuous variations and molar-ratio as Fe(III): DCIPF = 1:4, the excitation and emission wavelengths were found to be 282 and 560 nm, respectively. There is a linear relationship in the range 4-24 ng x mL(-1) for Fe(III). This method is simple and rapid, and has been applied to the determination of trace iron in Chinese herbal medicine and tea samples with satisfactory results.

Tryptophan and kynurenine determination in human hair by liquid chromatography.

PubMed

Dario, Michelli F; Freire, Thamires Batello; Pinto, Claudinéia Aparecida Sales de Oliveira; Prado, María Segunda Aurora; Baby, André R; Velasco, Maria Valéria R

2017-10-15

Tryptophan, an amino acid found in hair proteinaceous structure is used as a marker of hair photodegradation. Also, protein loss caused by several chemical/physical treatments can be inferred by tryptophan quantification. Kynurenine is a photo-oxidation product of tryptophan, expected to be detected when hair is exposed mainly to UVB (290-320nm) radiation range. Tryptophan from hair is usually quantified directly as a solid or after alkaline hydrolysis, spectrofluorimetrically. However, these types of measure are not sufficiently specific and present several interfering substances. Thus, this work aimed to propose a quantification method for both tryptophan and kynurenine in hair samples, after alkali hydrolysis process, by using high-performance liquid chromatography (HPLC) with fluorimetric and UV detection. The tryptophan and kynurenine quantification method was developed and validated. Black, white, bleached and dyed (blond and auburn) hair tresses were used in this study. Tryptophan and kynurenine were separated within ∼9min by HPLC. Both black and white virgin hair samples presented similar concentrations of tryptophan, while bleaching caused a reduction in the tryptophan content as well as dyeing process. Unexpectedly, UV/vis radiation did not promote significantly the conversion of tryptophan into its photo-oxidation product and consequently, kynurenine was not detected. Thus, this works presented an acceptable method for quantification of tryptophan and its photooxidation metabolite kynurenine in hair samples. Also, the results indicated that bleaching and dyeing processes promoted protein/amino acids loss but tryptophan is not extensively degraded in human hair by solar radiation. Copyright © 2017 Elsevier B.V. All rights reserved.

Catalytic spectrofluorimetric determination of superoxide anion radical and superoxide dismutase activity using N, N-dimethylaniline as the substrate for horseradish peroxidase (HRP)

NASA Astrophysics Data System (ADS)

Tang, Bo; Wang, Yan; Chen, Zhen-zhen

2002-10-01

The coupled reaction of N, N-dimethylaniline (DMA) with 4-aminoantipyrine (4-AAP) using superoxide anion radical (O 2-) as oxidizing agent under the catalysis of horseradish peroxidase (HRP) was studied. Based on the reaction, O 2- produced by irradiating Vitamin B 2, (V B2) was spectrophotometricly determined at 554 nm. The linear range of this method was 1.8×10 -6-1.2×10 -4 mol l -1 with a detection limit of 5.3×10 -7 mol l -1. The effect of interferences on the determination of O 2- was investigated. The proposed method was successfully applied to the determination of superoxide dismutase (SOD) activity in human blood and mouse blood.

Single excitation-emission fluorescence spectrum (EEF) for determination of cetane improver in diesel fuel.

PubMed

Insausti, Matías; Fernández Band, Beatriz S

2015-04-05

A highly sensitive spectrofluorimetric method has been developed for the determination of 2-ethylhexyl nitrate in diesel fuel. Usually, this compound is used as an additive in order to improve cetane number. The analytical method consists in building the chemometric model as a first step. Then, it is possible to quantify the analyte with only recording a single excitation-emission fluorescence spectrum (EEF), whose data are introduced in the chemometric model above mentioned. Another important characteristic of this method is that the fuel sample was used without any pre-treatment for EEF. This work provides an interest improvement to fluorescence techniques using the rapid and easily applicable EEF approach to analyze such complex matrices. Exploding EEF was the key to a successful determination, obtaining a detection limit of 0.00434% (v/v) and a limit of quantification of 0.01446% (v/v). Copyright © 2015 Elsevier B.V. All rights reserved.

New highly sensitive and selective fluorescent terbium complex for the detection of aluminium ions

NASA Astrophysics Data System (ADS)

Anwar, Zeinab M.; Ibrahim, Ibrahim A.; Kamel, Rasha M.; Abdel-Salam, Enas T.; El-Asfoury, Mahmoud H.

2018-02-01

A highly sensitive and selective spectrofluorimetric method has been developed for the rapid determination of aluminium ions. The method is based on the fluorescence enhancement of Tb complex with 3,4-dimetyl-thieno[2,3 b] thiophene-2,5-dicarboxylic acid (LN) after addition trace amount of aluminium ions. The fluorescence of the probe is monitored at the characteristic an emission wavelength of Tb3+ at 545 nm with excitation at 300 nm. Optimum detection was obtained in DMSO-H2O (2:8, v/v) and at pH 6.0 using MOPSO buffer. Under the optimum conditions linear calibration curves were obtained from 0.5 μ mol L-1 to 20 μ mol L-1 with detection limit of 0.1 μ mol L-1. Effect of interference of other ions was studied.

Analysis of quetiapine in human plasma using fluorescence spectroscopy

NASA Astrophysics Data System (ADS)

Mostafa, Islam M.; Omar, Mahmoud A.; Nagy, Dalia M.; Derayea, Sayed M.

2018-05-01

A simple and sensitive spectrofluorimetric method has been development for the assurance of quetiapine fumarate (QTF). The proposed method was utilized for measuring the fluorescence intensity of the yellow fluorescent product at 510 nm (λex 470 nm). The fluorescent product has resulted from the nucleophilic substitution reaction of QTF with 4-chloro-7-nitrobenzofurazane (NBD-Cl) in Mcllvaine buffer (pH 7.0). The diverse variables influencing the development of the reaction product were deliberately changed and optimized. The linear concentration range of the proposed method was of 0.2-2.0 μg ml-1.The limits of detection and quantitation were 0.05 and 0.17 μg ml-1, respectively. The proposed method was applied for the assurance of QTF in its tablets without interference from basic excipients. In addition, the proposed method was used for in vitro analysis of the QTF in spiked human plasma, the percent mean recovery was (n = 3) 98.82 ± 1.484%.

Assessing FRET using Spectral Techniques

PubMed Central

Leavesley, Silas J.; Britain, Andrea L.; Cichon, Lauren K.; Nikolaev, Viacheslav O.; Rich, Thomas C.

2015-01-01

Förster resonance energy transfer (FRET) techniques have proven invaluable for probing the complex nature of protein–protein interactions, protein folding, and intracellular signaling events. These techniques have traditionally been implemented with the use of one or more fluorescence band-pass filters, either as fluorescence microscopy filter cubes, or as dichroic mirrors and band-pass filters in flow cytometry. In addition, new approaches for measuring FRET, such as fluorescence lifetime and acceptor photobleaching, have been developed. Hyperspectral techniques for imaging and flow cytometry have also shown to be promising for performing FRET measurements. In this study, we have compared traditional (filter-based) FRET approaches to three spectral-based approaches: the ratio of acceptor-to-donor peak emission, linear spectral unmixing, and linear spectral unmixing with a correction for direct acceptor excitation. All methods are estimates of FRET efficiency, except for one-filter set and three-filter set FRET indices, which are included for consistency with prior literature. In the first part of this study, spectrofluorimetric data were collected from a CFP–Epac–YFP FRET probe that has been used for intracellular cAMP measurements. All comparisons were performed using the same spectrofluorimetric datasets as input data, to provide a relevant comparison. Linear spectral unmixing resulted in measurements with the lowest coefficient of variation (0.10) as well as accurate fits using the Hill equation. FRET efficiency methods produced coefficients of variation of less than 0.20, while FRET indices produced coefficients of variation greater than 8.00. These results demonstrate that spectral FRET measurements provide improved response over standard, filter-based measurements. Using spectral approaches, single-cell measurements were conducted through hyperspectral confocal microscopy, linear unmixing, and cell segmentation with quantitative image analysis. Results from these studies confirmed that spectral imaging is effective for measuring subcellular, time-dependent FRET dynamics and that additional fluorescent signals can be readily separated from FRET signals, enabling multilabel studies of molecular interactions. PMID:23929684

Assessing FRET using spectral techniques.

PubMed

Leavesley, Silas J; Britain, Andrea L; Cichon, Lauren K; Nikolaev, Viacheslav O; Rich, Thomas C

2013-10-01

Förster resonance energy transfer (FRET) techniques have proven invaluable for probing the complex nature of protein-protein interactions, protein folding, and intracellular signaling events. These techniques have traditionally been implemented with the use of one or more fluorescence band-pass filters, either as fluorescence microscopy filter cubes, or as dichroic mirrors and band-pass filters in flow cytometry. In addition, new approaches for measuring FRET, such as fluorescence lifetime and acceptor photobleaching, have been developed. Hyperspectral techniques for imaging and flow cytometry have also shown to be promising for performing FRET measurements. In this study, we have compared traditional (filter-based) FRET approaches to three spectral-based approaches: the ratio of acceptor-to-donor peak emission, linear spectral unmixing, and linear spectral unmixing with a correction for direct acceptor excitation. All methods are estimates of FRET efficiency, except for one-filter set and three-filter set FRET indices, which are included for consistency with prior literature. In the first part of this study, spectrofluorimetric data were collected from a CFP-Epac-YFP FRET probe that has been used for intracellular cAMP measurements. All comparisons were performed using the same spectrofluorimetric datasets as input data, to provide a relevant comparison. Linear spectral unmixing resulted in measurements with the lowest coefficient of variation (0.10) as well as accurate fits using the Hill equation. FRET efficiency methods produced coefficients of variation of less than 0.20, while FRET indices produced coefficients of variation greater than 8.00. These results demonstrate that spectral FRET measurements provide improved response over standard, filter-based measurements. Using spectral approaches, single-cell measurements were conducted through hyperspectral confocal microscopy, linear unmixing, and cell segmentation with quantitative image analysis. Results from these studies confirmed that spectral imaging is effective for measuring subcellular, time-dependent FRET dynamics and that additional fluorescent signals can be readily separated from FRET signals, enabling multilabel studies of molecular interactions. © 2013 International Society for Advancement of Cytometry. Copyright © 2013 International Society for Advancement of Cytometry.

Spectrofluorimetric and Potentiometric Determination of Acidity Constants of 4-(4'-Acetyloxy-3'-Methoxybenzylidene)-5-Oxazolone Derivatives.

PubMed

Taskiran, Derya Topkaya; Urut, Gulsiye Ozturk; Ayata, Sevda; Alp, Serap

2017-03-01

4-(4'-acetyloxy-3'-methoxybenzylidene)-5-oxazolone fluorescent molecules bearing four different aryl groups attached to the 2-position of 5-oxazolone ring have been investigated by spectrophotometric and potentiometric techniques in solution media. The acidity constants (pKa) of the fluorescent molecules were precisely determined in acetone, acetonitrile, dimethylformamide and in 1:1 mixture of toluene-isopropanol. The studied derivatives were titrated with tetrabutylammonium hydroxide and non-aqueous perchloric acid by scanning the basic and acidic region of the pH scale. A computerizable derivative method was used in order to descript precisely the end point and pKa values. The molecules investigated performed well-shaped and stoichiometric potentiometric titration curves.

Analytical investigation of the influence of ornidazole on the native protein fluorescence.

PubMed

Ali, Hazim M; El-Hashemy, Mohammed A

2018-05-03

A novel spectrofluorimetric method for the determination of ornidazole (ORN) in pure form and dosage forms was developed based on the influence of ORN on the native fluorescence of bovine serum albumin (BSA) in a stimulated physiological environment. The obtained data reveal that the presence of ORN has a strong quenching effect on the fluorescence of BSA through both a dynamic and a static process. The parameters of the binding of ORN to BSA were calculated at different temperatures. Thermodynamic parameters values suggest a role of electrostatic and hydrophobic forces in the binding of ORN to BSA. The investigated method for the determination of ORN is accurate, precise and sensitive with a detection limit of 0.106 μg/mL and a quantification limit of 0.353 μg/mL. The quenching method was applied successfully in the determination of ORN in pure form and dosage forms. Copyright © 2018 Elsevier B.V. All rights reserved.

A model to estimate the relative position of sites for ligands in serum albumins

NASA Astrophysics Data System (ADS)

Motta, Art Adriel Emidio de Araújo; Grassini, Maria Carolina Vilela; Cortez, Célia Martins; Silva, Dilson

2017-11-01

In this work, we present a mathematical-computational model developed to estimate the relative position of ligand binding sites in HSA and BSA, based on the theory of fluorescence quenching, considering the molecular and spectrofluorimetric differences and similarities between these two albumins. Albumin is the largest and the most abundant serum protein in vertebrates. The ability to bind xenobiotics makes albumin important to the bioavailability and effectiveness of drugs.

Determination of adenosine disodium triphosphate (ATP) using oxytetracycline-Eu 3+ as a fluorescence probe by spectrofluorimetry

NASA Astrophysics Data System (ADS)

Hou, Faju; Miao, Yanhong; Jiang, Chongqiu

2005-10-01

A new spectrofluorimetric method was developed for determination of adenosine disodium triphosphate (ATP). We studied the interactions between oxytetracycline (OTC)-Eu 3+ complex and adenosine disodium triphosphate (ATP) by using UV-vis absorption and fluorescence spectra. Using oxytetracycline (OTC)-Eu 3+ as a fluorescence probe, under the optimum conditions, ATP can remarkably enhance the fluorescence intensity of the OTC-Eu 3+ complex at λ = 612 nm and the enhanced fluorescence intensity of Eu 3+ ion is in proportion to the concentration of ATP. Optimum conditions for the determination of ATP were also investigated. The linear ranges for ATP are 8.00 × 10 -8-1.50 × 10 -6 mol L -1 with detection limits of 2.67 × 10 -9 mol L -1. This method is simple, practical and relatively free interference from coexisting substances and can be successfully applied to determination of ATP in samples. The mechanism of fluorescence enhancement between oxytetracycline (OTC)-Eu 3+ complex and ATP was also studied.

Using the Antenna Effect as a Spectroscopic Tool; Photophysics and Solution Thermodynamics of the Model Luminescent Hydroxypyridonate Complex [EuIII(3,4,3-LI(1,2-HOPO))]-

PubMed Central

Abergel, Rebecca J.; D'Aléo, Anthony; Leung, Clara Ng Pak; Shuh, David K.; Raymond, Kenneth N.

2009-01-01

While widely used in bioassays, the spectrofluorimetric method described here uses the antenna effect as a tool to probe the thermodynamic parameters of ligands that sensitize lanthanide luminescence. The Eu3+ coordination chemistry, solution thermodynamic stability and photophysical properties of the spermine-based hydroxypyridonate octadentate chelator 3,4,3-LI(1,2-HOPO) are reported. The complex [EuIII(3,4,3-LI(1,2-HOPO))]- luminesces with a long lifetime (805 μs) and a quantum yield of 7.0% in aqueous solution, at pH 7.4. These remarkable optical properties were exploited to determine the high (and proton-independent) stability of the complex (log β110 = 20.2(2)) and to define the influence of the ligand scaffold on the stability and photophysical properties. PMID:19902920

A highly selective and sensitive Tb3+-acetylacetone photo probe for the assessment of acetazolamide in pharmaceutical and serum samples

NASA Astrophysics Data System (ADS)

Youssef, A. O.

2018-04-01

A novel, simple, sensitive and selective spectrofluorimetric method was developed for the determination of Acetazolamide in pharmaceutical tablets and serum samples using photo probe Tb3+-ACAC. The Acetazolamide can remarkably quench the luminescence intensity of Tb3+-ACAC complex in DMSO at pH 6.8 and λex = 350 nm. The quenching of luminescence intensity of Tb3+-ACAC complex especially the electrical band at λem = 545 nm is used for the assessment of Acetazolamide in the pharmaceutical tablet and serum samples. The dynamic range found for the determination of Acetazolamide concentration is 4.49 × 10-9-1.28 × 10-7 mol L-1, and the limit of detection (LOD) and limit of quantification (LOQ) are (4.0 × 10-9 and 1.21 × 10-8) mol L-1, respectively.

[Urinary excretion of catecholamines in obese subjects and in diabetics (author's transl)].

PubMed

Giorgino, R; Nardelli, G M; Scardapane, R

1976-03-01

95 obese subjects, 40 diabetics and 22 normal controls were investigated. The weight of all obese subjects was at least 20% higher than the ideal weight. Catecholamine excretion was determined a few days after hospitalization to minimize the influence of environmental changes. Spectrofluorimetric estimation of adrenaline and noradrenaline in the urine was carried out according to the method of von Euler and Lihajko. Statistical analysis of the results showed a significant increase in both adrenaline and noradrenaline excretion in the group of obeses subjects compared with the diabetics. The increased catecholamine excretion may represent the response of the adrenal medulla to the stress of the disease. Such an increase may be responsible for perpheral insulin resistence and hence acts as a diabetogenic factor. The results obtained emphasize the influence of catecholamines on insulin responsiveness, possibly constituting a major contribution to the diabetic state.

Intracellular magnesium detection by fluorescent indicators.

PubMed

Trapani, Valentina; Schweigel-Röntgen, Monika; Cittadini, Achille; Wolf, Federica I

2012-01-01

Magnesium is essential for a wide variety of biochemical reactions and physiological functions, but its regulatory mechanisms (both at the cellular and at the systemic level) are still poorly characterized. Not least among the reasons for this gap are the technical difficulties in sensing minor changes occurring over a high background concentration. Specific fluorescent indicators are highly sensitive tools for dynamic evaluation of intracellular magnesium concentration. We herein discuss the main criteria to consider when choosing a magnesium-specific fluorescent indicator and provide examples among commercial as well as developmental sensors. We focus on spectrofluorimetric approaches to quantify Mg(2+) concentration in cell or mitochondria suspensions, and on imaging techniques to detect intracellular magnesium distribution and fluxes by live microscopy, reporting a detailed description of standard protocols for each method. The general guidelines we provide should be applicable to specific issues by any researcher in the field. Copyright © 2012 Elsevier Inc. All rights reserved.

Level of endogenous formaldehyde in maple syrup as determined by spectrofluorimetry.

PubMed

Lagacé, Luc; Guay, Stéphane; Martin, Nathalie

2003-01-01

The level of endogenous formaldehyde in maple syrup was established from a large number (n = 300) of authentic maple syrup samples collected during 2000 and 2001 in the province of Quebec, Canada. The average level of formaldehyde from these authentic samples was measured at 0.18 mg/kg in 2000 and 0.28 mg/kg in 2001, which is lower than previously published. These average values can be attributed to the improved spectrofluorimetric method used for the determination. However, the formaldehyde values obtained demonstrate a relatively large distribution with maximums observed at 1.04 and 1.54 mg/kg. These values are still under the maximum tolerance level of 2.0 mg/kg paraformaldehyde pesticide residue. Extensive heat treatment of maple syrup samples greatly enhanced the formaldehyde concentration of the samples, suggesting that extensive heat degradation of the sap constituents during evaporation could be responsible for the highest formaldehyde values in maple syrup.

Analysis of limette and bergamot distilled essential oils by HPLC.

PubMed

Buiarelli, Francesca; Cartoni, Giampaolo; Coccioli, Franco; Jasionowska, Renata; Mazzarino, Monica

2002-04-01

This work examines the distilled essential oils of limette and bergamot in order to assess the presence of low volatile substances such as coumarins (bergapten) which, being toxic, must be eliminated before using these oils in the food industry. The quantitative determination of coumarins was carried out by spectrofluorimetric detection. The substances present in the chromatograms, obtained by HPLC with UV detection at 254 nm, were then identified. Moreover, a new coumarin that is present in small quantities was identified using HPLC-MS.

[Determination of benzo(alpha)pyrene in food with microwave-assisted extraction].

PubMed

Zhou, Na; Luo, He-Dong; Li, Na; Li, Yao-Qun

2014-03-01

Coupling derivative technique and constant-energy synchronous fluorescence scanning technique, a method of determining benzo[alpha] pyrene in foods by second derivative constant-energy synchronous spectrofluorimetry after microwave-assisted treatment of samples was established using domestic microwave oven. The main factors of influencing the efficiency of microwave extraction were discussed, including the extraction solvent types and amounts, the microwave extraction time, microwave radiation power and cooling time. And the comparison with ultrasonic extraction was made. Low-fat food samples, which were just microwave-extracted with mixed-solvents, could be analyzed immediately by the spectrofluorimetric technique. For high-fat food samples, microwave-assisted saponification and extraction were made at the same time, thus simplifying operation steps and reducing sample analysis time. So the whole sample analysis process could be completed within one hour. This method was simple, rapid and inexpensive. In consequence, it was applied to determine benzo(a)pyrene in food with good reproducibility and the recoveries of benzo(alpha) pyrene ranged from 90.0% to 105.0% for the low fat samples and 83.3% to 94.6% for high-fat samples.

Automated laser spectrofluorimeter for monitoring of myocardial metabolism

NASA Astrophysics Data System (ADS)

Popov, A. Yu.; Salmin, V. V.; Fursov, A. A.; Stepanenko, A. V.; Sokolovich, A. G.; Salmina, A. B.; Rebenkova, A. A.; Makarov, R. A.; Provorov, A. S.

2006-09-01

Methods of optical biopsy have a series of advantages before other methods of clinical diagnostics. The high accuracy of received results enables registration even small change of concentration of substances, and the opportunity of remote registration makes methods optical biopsy by an optimum means for noninvasive methods of diagnostics in medicine. The method of the fluorescent analysis allows to investigate dynamics of changes of a functional condition of organs and tissue in norm and pathologies, called by the various factors (an inflammation, ischemia, degenerative changes). Bring the results of development of expiremental setup for the laser fluorescent analysis of physiological and functional condition of various organs and tissue of organism. In expiremental setup was used pulse UF nitric laser with length of wave generation = 337 nm. For delivery of radiation to tissue, and, also, collection of a radiation of fluorescence were used various optic fiber scheme. The expiremental setup includes automated tunable monochromator and ADC, receiving a signal from photomultiplier tube. Driving of all blocks and processing of results is realize on IBM-compatible computer with the appropriate software. Was used the synchronous detecting for reducing of a background signal. Myocard at surgical introoperation by an accompanied condition of sharp ischemia was researches on these expiremental setup. Spectrofluorimetric criteria of an estimation of a condition of viscus at peritonitis were development.

Determination of human albumin in serum and urine samples by constant-energy synchronous fluorescence method.

PubMed

Madrakian, Tayyebeh; Bagheri, Habibollah; Afkhami, Abbas

2015-08-01

A sensitive spectrofluorimetric method using constant-energy synchronous fluorescence technique is proposed for the determination of human albumin without separation. In this method, no reagent was used for enhancement of the fluorescence signal of albumin in the solution. Effects of some parameters, such as energy difference between excitation and emission monochromators (ΔE), emission and excitation slit widths and scan rate of wavelength were studied and the optimum conditions were established. For this purpose factorial design and response surface method were employed for optimization of the effective parameters on the fluorescence signal. The results showed that the scan rate of the wavelength has no significant effect on the analytical signal. The calibration curve was linear in the range 0.1-220.0 µg mL(-1) of albumin with a detection limit of 7.0 × 10(-3)  µg mL(-1). The relative standard deviations (RSD) for six replicate measurements of albumin were calculated as 2.2%, 1.7% and 1.3% for 0.5, 10.0 and 100.0 µg mL(-1) albumin, respectively. Furthermore the proposed method has been employed for the determination of albumin in human serum and urine samples. Copyright © 2014 John Wiley & Sons, Ltd.

Determination of fluoxetine in pharmaceutical and biological samples based on the silver nanoparticle enhanced fluorescence of fluoxetine-terbium complex.

PubMed

Lotfi, Ali; Manzoori, Jamshid L

2016-11-01

In this study, a simple and sensitive spectrofluorimetric method is presented for the determination of fluoxetine based on the enhancing effect of silver nanoparticles (AgNPs) on the terbium-fluoxetine fluorescence emission. The AgNPs were prepared by a simple reduction method and characterized by UV-Vis spectroscopy and transmission electron microscopy. It was indicated that these AgNPs have a remarkable amplifying effect on the terbium-sensitized fluorescence of fluoxetine. The effects of various parameters such as AgNP and Tb 3+ concentration and the pH of the media were investigated. Under obtained optimal conditions, the fluorescence intensity of the terbium-fluoxetine-AgNP system was enhanced linearly by increasing the concentration of fluoxetine in the range of 0.008 to 19 mg/L. The limit of detection (b + 3s) was 8.3 × 10 -4 mg/L. The interference effects of common species found in real samples were also studied. The method had good linearity, recovery, reproducibility and sensitivity, and was satisfactorily applied for the determination of fluoxetine in tablet formulations, human urine and plasma samples. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

Phosphorus dendrimers and photodynamic therapy. Spectroscopic studies on two dendrimer-photosensitizer complexes: Cationic phosphorus dendrimer with rose bengal and anionic phosphorus dendrimer with methylene blue.

PubMed

Dabrzalska, Monika; Zablocka, Maria; Mignani, Serge; Majoral, Jean Pierre; Klajnert-Maculewicz, Barbara

2015-08-15

Dendrimers due to their unique architecture may play an important role in drug delivery systems including chemotherapy, gene therapy and recently, photodynamic therapy as well. We investigated two dendrimer-photosensitizer systems in context of potential use of these systems in photodynamic therapy. The mixtures of an anionic phosphorus dendrimer of the second generation and methylene blue were studied by UV-vis spectroscopy while that of a cationic phosphorus dendrimer (third generation) and rose bengal were investigated by spectrofluorimetric methods. Spectroscopic analysis of these two systems revealed the formation of dendrimer-photosensitizer complexes via electrostatic interactions as well as π stacking. The stoichiometry of the rose bengal-cationic dendrimer complex was estimated to be 7:1 and 9:1 for the methylene blue-anionic dendrimer complex. The results suggest that these polyanionic or polycationic phosphorus dendrimers can be promising candidates as carriers in photodynamic therapy. Copyright © 2015 Elsevier B.V. All rights reserved.

Novel fluorimetric assay of trace analysis of epinephrine in human serum

NASA Astrophysics Data System (ADS)

Adeniyi, William K.; Wright, Ashleigh R.

2009-12-01

A simple, rapid, and sensitive spectrofluorimetric technique for the microdetermination of epinephrine in human serum is described. The investigation shows that trace amounts of epinephrine, antidepressant of clinical importance, can be determined without the conventional derivatization or use of fluorophores by diazotization. The method is based on the optimization of experimental parameters, such as pH, temperature, careful selection of excitation and emission wavelengths and on the use of anionic surfactant, sodium dodecyl sulfate (SDS), to enhance sensitivity. The measurement was carried out at 360 nm with excitation at 286 nm. Under the optimum conditions, a linear relationship was obtained between the fluorescence intensity and epinephrine concentration in the range of 0.10 and 1.0 μg/mL; the correlation coefficient and detection limit are 0.9953 and 0.05 μg/mL, respectively. Recovery tests indicated an efficiency of 95.5-98.7% by using known amounts of epinephrine spiked with human serum.

Comment: CAPN10 alleles are associated with polycystic ovary syndrome.

PubMed

Gonzalez, Alejandro; Abril, Eduardo; Roca, Alfredo; Aragón, Maria José; Figueroa, Maria José; Velarde, Pilar; Royo, José Luis; Real, Luis Miguel; Ruiz, Agustín

2002-08-01

Polycystic ovary syndrome (PCOS) is characterized by chronic anovulation infertility, hyperandrogenemia, and frequently insulin resistance. This study investigated whether polymorphisms in the CAPN10 gene are related with PCOS etiology. The allelic frequencies and genotypes of CAPN10 polymorphisms UCSNP-44, 43, 19, and 63 were determined in 55 well characterized women with polycystic ovaries and 93 unrelated healthy controls using spectrofluorimetric analyses and real-time PCR. Our data indicate that CAPN10 UCSNP-44 allele is associated with PCOS in the Spanish population (P = 0.01). These results support a role of Calpain 10 gene in PCOS susceptibility in humans.

Concentrations of cadmium and selected essential elements in malignant large intestine tissue

PubMed Central

Dziki, Adam; Kilanowicz, Anna; Sapota, Andrzej; Duda-Szymańska, Joanna; Daragó, Adam

2015-01-01

Introduction Colorectal cancer is one of the most common cancers worldwide. Incidence rates of large intestine cancer indicate a role of environmental and occupational factors. The role of essential elements and their interaction with toxic metals can contribute to the explanation of a complex mechanism by which large intestine cancer develops. Bearing this in mind, determining the levels of essential and toxic elements in tissues (organs), as well as in body fluids, seems to shed light on their role in the mode of action in malignant disease. Aim Determination of the levels of cadmium, zinc, copper, selenium, calcium, magnesium, and iron in large intestine malignant tissue. Material and methods Two intraoperative intestine sections were investigated: one from the malignant tissue and the other one from the normal tissue, collected from each person with diagnosed large intestine cancer. Cadmium, zinc, copper, calcium, magnesium, and iron levels were determined with atomic absorption spectrometry, and selenium levels by spectrofluorimetric method. Results The levels of copper, selenium, and magnesium were higher in the malignant than in normal tissues. In addition, the zinc/copper and calcium/magnesium relationship was altered in malignant tissue, where correlations were lower compared to non-malignant tissue. Conclusions The results seems to demonstrate disturbed homeostasis of some essential elements. However, it is hard to confirm their involvement in the aetiology of colorectal cancer. PMID:27110307

Sorptive thin film microextraction followed by direct solid state spectrofluorimetry: A simple, rapid and sensitive method for determination of carvedilol in human plasma.

PubMed

Karimi, Shima; Talebpour, Zahra; Adib, Noushin

2016-06-14

A poly acrylate-ethylene glycol (PA-EG) thin film is introduced for the first time as a novel polar sorbent for sorptive extraction method coupled directly to solid-state spectrofluorimetry without the necessity of a desorption step. The structure, polarity, fluorescence property and extraction performance of the developed thin film were investigated systematically. Carvedilol was used as the model analyte to evaluate the proposed method. The entire procedure involved one-step extraction of carvedilol from plasma using PA-EG thin film sorptive phase without protein precipitation. Extraction variables were studied in order to establish the best experimental conditions. Optimum extraction conditions were the followings: stirring speed of 1000 rpm, pH of 6.8, extraction temperature of 60 °C, and extraction time of 60 min. Under optimal conditions, extraction of carvedilol was carried out in spiked human plasma; and the linear range of calibration curve was 15-300 ng mL(-1) with regression coefficient of 0.998. Limit of detection (LOD) for the method was 4.5 ng mL(-1). The intra- and inter-day accuracy and precision of the proposed method were evaluated in plasma sample spiked with three concentration levels of carvedilol; yielding a recovery of 91-112% and relative standard deviation of less than 8%, respectively. The established procedure was successfully applied for quantification of carvedilol in plasma sample of a volunteer patient. The developed PA-EG thin film sorptive phase followed by solid-state spectrofluorimetric method provides a simple, rapid and sensitive approach for the analysis of carvedilol in human plasma. Copyright © 2016 Elsevier B.V. All rights reserved.

Estimation of individual sennosides in plant materials and marketed formulations by an HPTLC method.

PubMed

Shah, S A; Ravishankara, M N; Nirmal, A; Shishoo, C J; Rathod, I S; Suhagia, B N

2000-04-01

Senna is a well-known drug, used in the Ayurvedic and Allopathic systems of medicine, and is a treatment for constipation. The purgative action of senna and its formulations is due to the presence of sennosides A and B. An HPTLC method has been developed for the determination of individual sennosides (A, B, C, D) without any derivatization in marketed formulations (three tablet formulations, two granule formulations and one liquid formulation) and plant materials (senna leaf and pod). The methanolic solution of a sample was applied on a pre-coated silica gel G60 F254 TLC plate (E. Merck.) and was developed using n-propanol : ethyl acetate : water : glacial acetic acid (3 : 3 : 2 : 0.1 v/v) as the mobile phase. The relative band speeds (Rf values) obtained were 0.35, 0.25, 0.61, 0.46 for sennosides A, B, C and D, respectively. The densitometric response was monitored at 366nm. Calibration curves were found to be linear in the concentration ranges 193-1356, 402-2817, 71-497 and 132-927 ng per spot for sennosides A, B, C, and D, respectively. The correlation coefficients were found to be 0.9978, 0.9987, 0.9939 and 0.9983 respectively for sennosides A, B, C and D. The result obtained with the HPTLC method for total sennoside content was compared with the results using the pharmacopoeial methods (spectrophotometric (British Pharmacopoeia) and spectrofluorimetric (United States Pharmacopeia) using the 'F' test). The results revealed no significant difference in the three different methods for estimation of total sennoside. The proposed HPTLC method was found to be simple, specific, precise, accurate and rapid. It can be used for routine quality control of sennosides or senna-containing formulations for individual sennosides.

A rapid and sensitive assay for determination of doxycycline using thioglycolic acid-capped cadmium telluride quantum dots

NASA Astrophysics Data System (ADS)

Tashkhourian, Javad; Absalan, Ghodratollah; Jafari, Marzieh; Zare, Saber

2016-01-01

A rapid, simple and inexpensive spectrofluorimetric sensor for determination of doxycycline based on its interaction with thioglycolic acid-capped cadmium telluride quantum dots (TGA/CdTe QDs) has been developed. Under the optimum experimental conditions, the sensor exhibited a fast response time of <10 s. The results revealed that doxycycline could quench the fluorescence of TGA/CdTe QDs via electron transfer from the QDs to doxycycline through a dynamic quenching mechanism. The sensor permitted determination of doxycycline in a concentration range of 1.9 × 10-6-6.1 × 10-5 mol L-1 with a detection limit of 1.1 × 10-7 mol L-1. The sensor was applied for determination of doxycycline in honey and human serum samples.

Spectrofluorimetric determination of melatonin in kernels of four different Pistacia varieties after ultrasound-assisted solid-liquid extraction.

PubMed

Oladi, Elham; Mohamadi, Maryam; Shamspur, Tayebeh; Mostafavi, Ali

2014-11-11

Melatonin is normally consumed to regulate the body's biological cycle. However it also has therapeutic properties, such as anti-tumor, anti-aging and protects the immune system. There are some reports on the presence of melatonin in edible kernels such as walnuts, but the extraction of melatonin from pistachio kernels is reported here for the first time. For this, the methanolic extract of pistachio kernels was exposed to gas chromatography/mass spectrometry analysis which confirmed the presence of melatonin. A fluorescence-based method was applied for the determination of melatonin in different extracts. When excited at λ=275 nm, the fluorescence emission intensity of melatonin was measured at λ=366 nm. Ultrasound-assisted solid-liquid extraction was used for the extraction of melatonin from pistachio kernels prior to fluorimetric determination. To achieve the highest extraction recovery, the main parameters affecting the extraction efficiency such as extracting solvent type and volume, temperature, sonication time and pH were evaluated. Under the optimized conditions, a linear dependence of fluorescence intensity on melatonin concentration was observed in the range of 0.0040-0.160 μg mL(-1), with a detection limit of 0.0036 μg mL(-1). This method was applied successfully for measuring and comparing the melatonin content in the kernels of four different varieties of Pistacia including Ahmad Aghaei, Akbari, Kalle Qouchi and Fandoghi. In addition, the results obtained were compared with those obtained using GC/MS. A good agreement was observed indicating the reliability of the proposed method. Copyright © 2014 Elsevier B.V. All rights reserved.

Spectrofluorimetric determination of melatonin in kernels of four different Pistacia varieties after ultrasound-assisted solid-liquid extraction

NASA Astrophysics Data System (ADS)

Oladi, Elham; Mohamadi, Maryam; Shamspur, Tayebeh; Mostafavi, Ali

2014-11-01

Melatonin is normally consumed to regulate the body's biological cycle. However it also has therapeutic properties, such as anti-tumor, anti-aging and protects the immune system. There are some reports on the presence of melatonin in edible kernels such as walnuts, but the extraction of melatonin from pistachio kernels is reported here for the first time. For this, the methanolic extract of pistachio kernels was exposed to gas chromatography/mass spectrometry analysis which confirmed the presence of melatonin. A fluorescence-based method was applied for the determination of melatonin in different extracts. When excited at λ = 275 nm, the fluorescence emission intensity of melatonin was measured at λ = 366 nm. Ultrasound-assisted solid-liquid extraction was used for the extraction of melatonin from pistachio kernels prior to fluorimetric determination. To achieve the highest extraction recovery, the main parameters affecting the extraction efficiency such as extracting solvent type and volume, temperature, sonication time and pH were evaluated. Under the optimized conditions, a linear dependence of fluorescence intensity on melatonin concentration was observed in the range of 0.0040-0.160 μg mL-1, with a detection limit of 0.0036 μg mL-1. This method was applied successfully for measuring and comparing the melatonin content in the kernels of four different varieties of Pistacia including Ahmad Aghaei, Akbari, Kalle Qouchi and Fandoghi. In addition, the results obtained were compared with those obtained using GC/MS. A good agreement was observed indicating the reliability of the proposed method.

New luminescent bioprobes Eu(lll)-phloroglucinol derivatives and their spectrofluorimetric, electrochemical interactions with nucleotides and DNA.

PubMed

Azab, Hassan Ahmed; Anwar, Zeinab M; Abdel-Salam, Enas T; el-Sayed-Sebak, Mahmoud

2012-01-01

Two new ligands derived from phloroglucinol 2-{[(4-methoxy benzoyl)oxy]} methyl benzoic acid[L1] and 2-{[(4-methyl benzoyl)oxy] methyl} benzoic acid[L2] were synthesized. The solid complex Eu(III)-L2 has been synthesised and characterized by elemental analysis, UV and IR spectra. The reaction of Eu(III) with the two synthesized ligands has been investigated in I = 0.1 mol dm(-3) p-toluene sulfonate by cyclic voltammetry and square wave voltammetry. The reaction of Eu (III)-L1 and Eu (III)-L2 binary complexes with nucleotide 5'-AMP, 5'-ADP, 5'-ATP, 5'- GMP, 5'-IMP, and 5'-CMP has been investigated using UV, fluorescence and electrochemical methods. The experimental conditions were selected such that self-association of the nucleotides and their complexes was negligibly small, that is, the monomeric complexes were studied. The interaction of the Eu(III)-L1 or L2 solid complexes with calf-thymus DNA has been investigated by fluorescence and electrochemical methods including cyclic voltammetery(CV), differential pulse polarography (DPP) and square wave voltammetry (SWV) on a glassy carbon electrode. The fluorescence intensity of Eu(III)-L2 complex was enhanced with the addition of DNA. Under optimal conditions in phosphate buffer pH 7.0 at 25 °C the linear range is 3-20 μM for calf thymus DNA (CT-DNA) and the corresponding determination limit is 1.8 μM.

Occupational exposure to coal tar pitch volatiles, benzo/a/pyrene and dust in tyre production.

PubMed

Rogaczewska, T; Ligocka, D

1994-01-01

Occupational exposure to coal tar pitch volatiles (CTPVs), benzo/a/pyrene (BaP) and dust was evaluated by means of individual measurements carried out in 80 workers and by stationary measurements on 16 work-posts in two divisions of the tyre producing plant. Dust and coal tar pitch volatiles concentrations in the air were determinated by the gravimetric method, measured, in the case of CPTVs, benzene-soluble fraction (BSF) with ultrasonic extraction. Benzo/a/pyrene analysis was performed using high performance liquid chromato-graphy (HPLC) with a spectrofluorimetric detector. It was found that nearly all personal sampling results for BaP were within the range < 4 divided by 142 ng/m3, except for the exposure of workers employed at weighing the raw materials (3,470-6,060 ng/m3) in the Semiproducts Division. Attention should be paid to the recorded CTPVs concentrations (benzene solubles). About 56% of the Vulcanizing Division workers and about 90% of the Semiproducts Division workers were exposed to these substances at concentrations of over 0.2 mg/m3 (hygienic standard for benzene solubles in USA). Exposure to dust (of high respirable fraction percentage > 90%) which exceeded the admissible value (4 mg/m3) was found mainly only in the workers of the Semiproducts Division at some work-posts.

Critical insight into the interaction of naringenin with human haemoglobin: A combined spectroscopic and computational modeling approaches

NASA Astrophysics Data System (ADS)

Maity, Subhajit; Chakraborty, Sandipan; Chakraborti, Abhay Sankar

2017-02-01

The present study demonstrates critical insight into the binding of a bioactive flavanone naringenin with normal human haemoglobin (NHb). Both spectrophotometric and spectrofluorimetric studies reveal that naringenin interacts with NHb. The binding affinity constant and number of binding sites appear to be approximately (1.5 ± 0.2) × 104 M-1 and 1, respectively. Static quenching seems to be an important factor in binding process, as evident from steady-state and time-resolved fluorescence spectroscopic studies. Far UV circular dichroism spectroscopy depicts that binding of naringenin to NHb causes no change in the secondary structure of the protein, which is also evident from Fourier transform infrared spectroscopic study. Free energy change (ΔG0) for naringenin-NHb interaction, determined by spectroscopic and isothermal calorimetric method, appears to be -5.67 kcal/mol and -6.90 kcal/mol, respectively, and is close to the docking energy -6.84 kcal/mol. Molecular docking suggests that naringenin binds near the cavity of the tetrameric heme protein, forming hydrogen bonds with surrounding amino acid residues. The binding site is away from the heme moieties, implicating naringenin binding does not affect the oxygen binding capacity of NHb, which makes the protein a suitable carrier of the flavonoid.

A rapid and sensitive assay for determination of doxycycline using thioglycolic acid-capped cadmium telluride quantum dots.

PubMed

Tashkhourian, Javad; Absalan, Ghodratollah; Jafari, Marzieh; Zare, Saber

2016-01-05

A rapid, simple and inexpensive spectrofluorimetric sensor for determination of doxycycline based on its interaction with thioglycolic acid-capped cadmium telluride quantum dots (TGA/CdTe QDs) has been developed. Under the optimum experimental conditions, the sensor exhibited a fast response time of <10s. The results revealed that doxycycline could quench the fluorescence of TGA/CdTe QDs via electron transfer from the QDs to doxycycline through a dynamic quenching mechanism. The sensor permitted determination of doxycycline in a concentration range of 1.9×10(-6)-6.1×10(-5)molL(-1) with a detection limit of 1.1×10(-7)molL(-1). The sensor was applied for determination of doxycycline in honey and human serum samples. Copyright © 2015 Elsevier B.V. All rights reserved.

Cytotoxic activity of proflavine diureas: synthesis, antitumor, evaluation and DNA binding properties of 1',1''-(acridin-3,6-diyl)-3',3''-dialkyldiureas.

PubMed

Kozurková, Mária; Sabolová, Danica; Janovec, Ladislav; Mikes, Jaromír; Koval', Ján; Ungvarský, Ján; Stefanisinová, Miroslava; Fedorocko, Peter; Kristian, Pavol; Imrich, Ján

2008-04-01

The synthesis of novel 1',1''-(acridin-3,6-diyl)-3',3''-dialkyldiureas was reported. Their biological activity to inhibit cell proliferation was assessed by a MTT assay on two cell lines, HeLa and HCT-116, at micromolar concentration. 1',1''-(Acridin-3,6-diyl)-3',3''-dihexyldiurea hydrochloride was active on a HCT-116 cell line with an IC(50) value of 3.1 microM. The interaction of these compounds with calf thymus DNA was investigated by a variety of spectroscopic techniques including UV-vis, fluorescence and CD spectroscopy. From spectrofluorimetric titrations, binding constants for the DNA-drug complexes were determined (K=0.9-4.2x10(5) M(-1)). Antiproliferative activity of synthesized derivatives might be related to their intercalation into DNA.

Uptake of the antileishmania drug tafenoquine follows a sterol-dependent diffusion process in Leishmania.

PubMed

Manzano, José Ignacio; Carvalho, Luis; García-Hernández, Raquel; Poveda, José Antonio; Ferragut, José Antonio; Castanys, Santiago; Gamarro, Francisco

2011-11-01

The present study was designed to elucidate the mechanism of tafenoquine uptake in Leishmania and its sterol dependence. Because tafenoquine is a fluorescent compound, spectrofluorimetric analysis allowed us to monitor its uptake by Leishmania promastigotes and intracellular amastigotes, and to evaluate the effect of temperature, energy and H+ gradient on drug entry. The influence of sterols on tafenoquine uptake in Leishmania parasites was determined in experiments using sterol-depleting agents such as methyl-β-cyclodextrin or cholesterol oxidase. Tafenoquine exhibited fast entry kinetics into Leishmania in an energy-independent, but pH- and temperature-dependent, non-saturable process. Furthermore, sterol depletion decreased tafenoquine uptake. These findings suggest that Leishmania takes up tafenoquine by a diffusion process and that decreases in membrane sterol content may induce a decrease in drug uptake.

Sequential determination of lead and cobalt in tap water and foods samples by fluorescence.

PubMed

Talio, María Carolina; Alesso, Magdalena; Acosta, María Gimena; Acosta, Mariano; Fernández, Liliana P

2014-09-01

In this work, a new procedure was developed for the separation and preconcentration of lead(II) and cobalt(II) in several water and foods samples. Complexes of metal ions with 8-hydroxyquinolein (8-HQ) were formed in aqueous solution. The proposed methodology is based on the preconcentration/separation of Pb(II) by solid-phase extraction using paper filter, followed by spectrofluorimetric determination of both metals, on the solid support and the filtered aqueous solution, respectively. The solid surface fluorescence determination was carried out at λem=455 nm (λex=385 nm) for Pb(II)-8-HQ complex and the fluorescence of Co(II)-8-HQ was determined in aqueous solution using λem=355 nm (λex=225 nm). The calibration graphs are linear in the range 0.14-8.03×10(4) μg L(-1) and 7.3×10(-2)-4.12×10(3) μg L(-1), for Pb(II) and Co(II), respectively, with a detection limit of 4.3×10(-2) and 2.19×10(-2) μg L(-1) (S/N=3). The developed methodology showed good sensitivity and adequate selectivity and it was successfully applied to the determination of trace amounts of lead and cobalt in tap waters belonging of different regions of Argentina and foods samples (milk powder, express coffee, cocoa powder) with satisfactory results. The new methodology was validated by electrothermal atomic absorption spectroscopy with adequate agreement. The proposed methodology represents a novel application of fluorescence to Pb(II) and Co(II) quantification with sensitivity and accuracy similar to atomic spectroscopies. Copyright © 2014 Elsevier B.V. All rights reserved.

A model based on spectrofluorimetry to study the interaction between glyphosate and serum albumin of Salminus brasiliensis

NASA Astrophysics Data System (ADS)

Escobar, Marta Araujo Cyrino; Cortez, Celia Martins; Silva, Dilson; Neto, Jayme da Cunha Bastos

2017-11-01

The aim of this work is to initiate an investigation on the albumin of Salminus brasiliensis (gold fish) as a biomarker of environmental actions of glyphosate. We started using a mathematical-computational model based on spectrofluorimetric measurements to study the interaction of glyphosate with gold fish albumin and human serum albumin. Salminus brasiliensis is a migratory freshwater fish species found in southern and central-western Brazil, mainly in the Prata river basin, where most of soybean plantations are set. Glyphosate is a very used herbicide in this type of crop. Differently from the organophosphorate methyl parathion, glyphosate does not form complex with HSA, and the quenching constants estimated for its binding with gold fish albumin at 20 °C and 25 °C is 1.3(± 0.3) × 104 / M e 2.5 (± 0.3) × 104 / M, respectively.

Analysis of epothilone B-induced cell death in normal ovarian cells.

PubMed

Rogalska, Aneta; Gajek, Arkadiusz; Marczak, Agnieszka

2013-12-01

We have investigated the mode of cell death induced by a new microtubule-stabilizing agent, epothilone B (EpoB, patupilone), and a clinically used medicine, paclitaxel (PTX), in normal ovarian cells. Using fluorescence microscopy, polyacrylamide gel electrophoresis preceding Western blot analysis, as well as spectrofluorimetric and colorimetric detection, we demonstrate that, compared to EpoB, PTX induced high time-dependent morphological and biochemical changes typical of apoptosis. Induction of apoptosis followed an early increase in p53 levels. Apoptosis reached its maximum at 24-48 h. At the same time, there was a significant increase in caspase-9 and -3 activity and PARP fragmentation, which suggests that an intrinsic path was involved. Apoptosis in MM14 cells was increased more by PTX than EpoB, and also induced more necrosis responsible for inflammation (1.4-fold) than EpoB. © 2013 International Federation for Cell Biology.

Interaction of sulpiride and serum albumin: Modeling from spectrofluorimetric data

NASA Astrophysics Data System (ADS)

Fragoso, Viviane Muniz da Silva; Silva, Dilson

2015-12-01

We have applied the fluorescence quenching modeling to study the process of interaction of sulpiride with human serum albumin (HSA) and bovine (BSA). Albumin is more abundant protein in blood and it emits fluorescence when excited by 260-295 nm. Sulpiride is an atypical antipsychotic used in the treatment of many psychiatric disorders. As sulpiride is fluorescent, we developed a mathematical model to analyzing the interaction of two fluorescent substances. This model was able to separate the albumin fluorescence from the quencher fluorescence. Results have shown that sulpiride quenches the fluorescence of both albumins by a static process, due to the complex formation drugalbumin. The association constants calculated for sulpiride-HSA was 2.20 (± 0.08) × 104 M-1 at 37° C, and 5.46 (± 0.20) × 104 M-1, 25 ° C, and the primary binding site to sulpiride in the albumin is located closer to the subdomain IB.

The influence of visible light and inorganic pigments on fluorescence excitation emission spectra of egg-, casein- and collagen-based painting media

NASA Astrophysics Data System (ADS)

Nevin, A.; Anglos, D.; Cather, S.; Burnstock, A.

2008-07-01

Spectrofluorimetric analysis of proteinaceous binding media is particularly promising because proteins employed in paintings are often fluorescent and media from different sources have significantly different fluorescence spectral profiles. Protein-based binding media derived from eggs, milk and animal tissue have been used for painting and for conservation, but their analysis using non-destructive techniques is complicated by interferences with pigments, their degradation and their low concentration. Changes in the fluorescence excitation emission spectra of films of binding media following artificial ageing to an equivalent of 50 and 100 years of museum lighting include the reduction of bands ascribed to tyrosine, tryptophan and Maillard reaction products and an increase in fluorescent photodegradation. Fluorescence of naturally aged paint is dependent on the nature of the pigment present and, with egg-based media, in comparison with un-pigmented films, emissions ascribed to amino acids are more pronounced.

Biophysical studies of interaction between hydrolysable tannins isolated from Oenothera gigas and Geranium sanguineum with human serum albumin.

PubMed

Sekowski, Szymon; Ionov, Maksim; Kaszuba, Mateusz; Mavlyanov, Saidmukhtar; Bryszewska, Maria; Zamaraeva, Maria

2014-11-01

Tannins, secondary plant metabolites, possess diverse biological activities and can interact with biopolymers such as lipids or proteins. Interactions between tannins and proteins depend on the structures of both and can result in changes in protein structure and activity. Because human serum albumin is the most abundant protein in plasma and responsible for interactions with important biological compounds (e.g. bilirubin) and proper blood pressure, therefore, it is very important to investigate reactions between HSA and tannins. This paper describes the interaction between human serum albumin (HSA) and two tannins: bihexahydroxydiphenoyl-trigalloylglucose (BDTG) and 1-O-galloyl-4,6-hexahydroxydiphenoyl-β-d-glucose (OGβDG), isolated from Geranium sanguineum and Oenothera gigas leafs, respectively. Optical (spectrofluorimetric) and chiral optical (circular dichroism) methods were used in this study. Fluorescence analysis demonstrated that OGβDG quenched HSA fluorescence more strongly than BDTG. Both OGβDG and BDTG formed complexes with albumin and caused a red shift of the fluorescence spectra but did not significantly change the protein secondary structure. Our studies clearly demonstrate that the tested tannins interact very strongly with human serum albumin (quenching constant K=88,277.26±407.04 M(-1) and K=55,552.67±583.07 M(-1) respectively for OGβDG and BDTG) in a manner depending on their chemical structure. Copyright © 2014 Elsevier B.V. All rights reserved.

Migration test of Bisphenol A from polycarbonate cups using excitation-emission fluorescence data with parallel factor analysis.

PubMed

Spagnuolo, M L; Marini, F; Sarabia, L A; Ortiz, M C

2017-05-15

Bisphenol A (BPA) is one of the most largely produced chemical in the world; it is used to make plastics and epoxy resins. The endocrine disruptor potential of BPA is well known, but recent researches suggest a relationship between chronic exposure to BPA, genotoxic activity and epigenetic modifications. The main source of exposure to BPA includes food contact materials (FCM). Thus simple and robust test methods are needed to improve the migration test of BPA. In this work, a non-separative, easy, fast and inexpensive spectrofluorimetric method based on the second order calibration of excitation-emission fluorescence matrices (EEMs) was proposed for the determination of BPA. For the first time, molecular fluorescence was used to identify unequivocally and quantify BPA. Trilinearity of the data tensor guarantees the uniqueness of the solution obtained through parallel factor analysis (PARAFAC), so one factor of the decomposition matches up with BPA even if other fluorophores are in the test sample. The effect of four experimental factors of the procedure on the figures of merit and the unequivocally identification was investigated by means of a D-optimal design and PARAFAC calibration. The method is linear and accurate in the range 0-720µgL -1 . The decision limit CCα and detection capability CCβ are 6.63µgL -1 and 18.85µgL -1 respectively (with probabilities of false positive and false negative fixed at 0.05). Finally the proposed method was applied to carry out a migration test from two polycarbonate cups, using 3% (w/v) acetic acid in aqueous solution as food simulant. The migrated amount of BPA was found to be 688.7µgL -1 (n=5) for the first cup and 710.5µgL -1 (n=4) for the second one, above the specific migration limit set by EFSA (European Food Safety Authority). Copyright © 2017 Elsevier B.V. All rights reserved.

DNA binding by a new metallointercalator that contains a proflavine group bearing a hanging chelating unit.

PubMed

Bazzicalupi, Carla; Bencini, Andrea; Bianchi, Antonio; Biver, Tarita; Boggioni, Alessia; Bonacchi, Sara; Danesi, Andrea; Giorgi, Claudia; Gratteri, Paola; Ingraín, Antonio Marchal; Secco, Fernando; Sissi, Claudia; Valtancoli, Barbara; Venturini, Marcella

2008-01-01

The new bifunctional molecule 3,6-diamine-9-[6,6-bis(2-aminoethyl)-1,6-diaminohexyl]acridine (D), which is characterised by both an aromatic moiety and a separate metal-complexing polyamine centre, has been synthesised. The characteristics of D and its ZnII complex ([ZnD]) (protonation and metal-complexing constants, optical properties and self-aggregation phenomena) have been analysed by means of NMR spectroscopy, potentiometric, spectrophotometric and spectrofluorimetric techniques. The equilibria and kinetics of the binding process of D and [ZnD] to calf thymus DNA have been investigated at I=0.11 M (NaCl) and 298.1 K by using spectroscopic methods and the stopped-flow technique. Static measurements show biphasic behaviour for both D-DNA and [ZnD]-DNA systems; this reveals the occurrence of two different binding processes depending on the polymer-to-dye molar ratio (P/D). The binding mode that occurs at low P/D values is interpreted in terms of external binding with a notable contribution from the polyamine residue. The binding mode at high P/D values corresponds to intercalation of the proflavine residue. Stopped-flow, circular dichroism and supercoiled-DNA unwinding experiments corroborate the proposed mechanism. Molecular-modelling studies support the intercalative process and evidence the influence of NH+...O interactions between the protonated acridine nitrogen atom and the oxygen atoms of the polyanion; these interactions play a key role in determining the conformation of DNA adducts.

Interaction of toxic azo dyes with heme protein: biophysical insights into the binding aspect of the food additive amaranth with human hemoglobin.

PubMed

Basu, Anirban; Kumar, Gopinatha Suresh

2015-05-30

A biophysical study on the interaction of the food colorant amaranth with hemoglobin was undertaken. Spectrophotometric and spectrofluorimetric studies proposed for an intimate binding interaction between the dye and the protein. The dye quenched the fluorescence of the protein remarkably and the mechanism of quenching was found to be static in nature. Synchronous fluorescence studies suggested that the polarity around the tryptophan residues was altered in the presence of amaranth whereas the polarity around tyrosine residues remained largely unaltered. 3D fluorescence, FTIR and circular dichroism results suggested that the binding reaction caused conformational changes in hemoglobin. The negative far-UV CD bands exhibited a significantly large decrease in magnitude in the presence of amaranth. From calorimetry studies it was established that the binding was driven by a large positive entropic contribution and a small but favorable enthalpy change. Copyright © 2015 Elsevier B.V. All rights reserved.

Chemodosimetric analysis in food-safety monitoring: design, synthesis, and application of a bimetallic Re(I)-Pt(II) complex for detection of dimethyl sulfide in foods.

PubMed

Chow, Cheuk-Fai; Gong, Fu-Wen; Gong, Cheng-Bin

2014-09-21

Detection of neutral biogenic sulfides plays a crucial role in food safety. A new heterobimetallic Re(I)-Pt(II) donor-acceptor complex--[Re(biq)(CO)3(CN)]-[Pt(DMSO)(Cl)2] (1, biq = 2,2'-biquinoline)--was synthesized and characterized. The X-ray crystallographic and photophysical data for 1 are reported in this study. Complex 1 indicated the luminescent chemodosimetric selectivity for dimethyl sulfide, which persisted even in the presence of a variety of interfering vapors, with a detection limit as low as 0.96 ppm. The binding constant (log K) of 1 toward dimethyl sulfide was 3.63 ± 0.03. The analyte selectivity of the complexes was found to be dependent on the ligand coordinated to the Re(I) center. Real samples (beef, chicken, and pork) were monitored real-time for gaseous dimethyl sulfide. Complex 1 shows a linear spectrofluorimetric response with increasing storage time of the meats at 30 °C.

Spectroscopic and spectrofluorimetric studies on the interaction of albendazole and trimethoprim with iodine

NASA Astrophysics Data System (ADS)

Ganesh, K.; Elango, K. P.

Raman, UV-vis, FT-IR, and fluorescence spectral techniques were employed to investigate the mechanism of interaction of albendazole (ALB) and trimethoprim (TMP) drugs with iodine. Interactions of ALB and TMP with iodine yields triiodide ion and its formation was confirmed by electronic and Raman spectra. The peaks appeared in Raman spectra of the isolated products are at around 145, 113 and 82 cm-1 are assigned to νas(I-I), νs(I-I) and δ(I3-) respectively, confirmed the presence of I3- ion. Formation constant (K), molar extinction coefficient (ɛ) and thermodynamic properties ΔH#, ΔS# and ΔG# were determined and discussed. Fluorescence quenching studies indicated that the interaction between the ALB, TMP with iodine are spontaneous and the TMP-iodine interaction is found to be stronger than that the other system. Solvent variation studies indicated that the binding constant increased with an increase in polarity of the medium.

Tissue distribution of selenium and effect of season and age on selenium content in roe deer from northwestern Poland.

PubMed

Pilarczyk, Bogumiła; Tomza-Marciniak, Agnieszka; Pilarczyk, Renata; Hendzel, Diana; Błaszczyk, Barbara; Bąkowska, Małgorzata

2011-06-01

The aim of the study was to compare selenium concentrations in different organs of roe deer from northwestern Poland. Samples of liver, kidneys, heart and lungs, collected from 74 roe deer shot during the hunting seasons of 2008-2009 in northwestern Poland, were studied. Selenium concentration in the organs was determined spectrofluorimetrically. Mean selenium concentration was 0.06 µg/g w.w. in the liver, 0.41 µg/g w.w. in the kidneys and 0.05 µg/g w.w. in the heart and lungs. Season had a significant effect on selenium concentration in the liver, kidneys, lungs and heart. In all the organs, the highest selenium concentration was found in spring and the lowest in autumn and winter. All animals studied were deficient in selenium. The low selenium concentration in the liver or heart can disturb their function, and in the future, it may be a factor contributing to the population decline of roe deer in the northwestern part of Poland.

Determination of josamycin residues in porcine tissues using high-performance liquid chromatography with pre-column derivatization and spectrofluorimetric detection.

PubMed

Leroy, P; Decolin, D; Nicolas, A; Archimbault, P

1994-12-01

A simple, selective and sensitive high-performance liquid chromatographic (HPLC) method has been developed for the measurement of josamycin residues in four porcine tissues (i.e., muscle, liver, kidney and fat). The sample preparation consisted of a homogenization step in an acetonitrile-10 mmol l-1 phosphate buffer mixture, pH 6.0 (35 + 65), centrifugation and a liquid-liquid extractive clean-up of the resulting supernatant with isooctane. Pre-column derivatization of josamycin was performed using cyclohexa-1,3-dione in ammonium acetate buffer, pH 5.0 (90 degrees C for 2 h). The derivative was chromatographed in an isocratic reversed-phase HPLC system. A LiChrospher RP 18 end-capped (5 microns) column was eluted with an acetonitrile-methanol-10 mmol l-1 phosphate buffer mixture, pH 6.0 (45 + 5 + 50). The capacity factor of the josamycin derivative was 17.5. Detection was achieved using spectrofluorimetry (lambda ex = 375 nm; lambda em = 450 nm). The structure of the derivative was assessed by using mass spectrometry. Full selectivity was obtained in the HPLC system versus other macrolide antibiotics (tylosin, spiramycin and erythromycin), aldehydes (formaldehyde, acetaldehyde and benzaldehyde) and endogenous compounds. Linearity and repeatability were tested. Correlation coefficients, for calibration curves in the range of 0.1-3.2 micrograms g-1, were greater than 0.999 for all tissues and the relative standard deviation (S(r)) was 4.9% (1.6 micrograms g-1; n = 6); recovery was higher than 88%.

Protolichesterinic acid enhances doxorubicin-induced apoptosis in HeLa cells in vitro.

PubMed

Brisdelli, Fabrizia; Perilli, Mariagrazia; Sellitri, Doriana; Bellio, Pierangelo; Bozzi, Argante; Amicosante, Gianfranco; Nicoletti, Marcello; Piovano, Marisa; Celenza, Giuseppe

2016-08-01

The aim of this study was to investigate the effect of protolichesterinic acid, a lichen secondary metabolite, on anti-proliferative activity of doxorubicin in three human cancer cell lines, HeLa, SH-SY5Y and K562 cells. The data obtained from MTT assays, performed on cells treated with protolichesterinic acid and doxorubicin alone and in combination, were analysed by the median-effect method as proposed by Chou and Talalay and the Bliss independence model. Apoptosis rate was evaluated by fluorescence microscopy, caspase-3, 8 and 9 activities were detected by spectrofluorimetric analysis and protein expression of Bim, Bid, Bax and Mcl-2 was analysed by Western blotting. The interaction of protolichesterinic acid with thioesterase domain of human fatty acid synthase (hFAS) was investigated by a molecular docking study. The in vitro activity of doxorubicin against HeLa cancer cell line, but not against SH-SY5Y and K562 cells, was synergically increased by protolichesterinic acid. The increased cytotoxicity caused by protolichesterinic acid in HeLa cells was due to a pro-apoptotic effect and was associated to caspase-3, 8 and 9 activation. The simultaneous treatment for 24h with protolichesterinic acid plus doxorubicin caused an increase of Bim protein expression and the appearance of cleaved form of Bid protein. The molecular modelling analysis showed that protolichesterinic acid seemed to behave as a competitive inhibitor of hFAS. These results suggest that protolichesterinic acid could be envisaged as an useful tool against certain types of tumor cells in combination with anticancer drugs. Copyright © 2016 Elsevier Inc. All rights reserved.

Spectrofluorimetric and molecular docking studies on the interaction of cyanidin-3-O-glucoside with whey protein, β-lactoglobulin.

PubMed

Cheng, Jing; Liu, Jian-Hua; Prasanna, Govindarajan; Jing, Pu

2017-12-01

The interaction of β-Lactoglobulin (β-Lg) with cyanidin-3-O-glucoside (C3G) was characterized using fluorescence, circular dichroism spectroscopy, and docking studies under physiological conditions. Fluorescence studies showed that β-Lg has a strong binding affinity for C3G via hydrophobic interaction with the binding constant, K a , of 3.14×10 4 M -1 at 298K. The secondary structure of β-Lg displayed an increase in the major structure of β-sheet upon binding with C3G, whereas a decrease in the minor structure of α-helix was also observed. In addition, evidenced by near UV-CD, the interaction also disrupted the environments of Trp residues. The molecular docking results illustrated that both hydrogen bonding and the hydrophobic interaction are involved as an acting force during the binding process. These results may contribute to a better understanding over the enhanced physicochemical proprieties of anthocyanins due to the complexation with milk proteins. Copyright © 2017 Elsevier B.V. All rights reserved.

Modulation of biogenic amines content by poly(propylene imine) dendrimers in rats.

PubMed

Ciepluch, Karol; Ziemba, Barbara; Janaszewska, Anna; Appelhans, Dietmar; Klajnert, Barbara; Bryszewska, Maria; Fogel, Wiesława Agnieszka

2012-09-01

Biogenic amines and polyamines participate in all vital organism functions, their levels being important function determinants. Studies were performed to check whether repeated administration of poly(propylene imine) (PPI) dendrimers, synthetic macromolecules with diaminobutane core, and peripheral primary amine groups, may influence the endogenous level of amines, as represented by the two of them: spermidine, a natural derivative of diaminobutane, and histamine. The experiment was carried out on Wistar rats. Fourth generation PPI dendrimer, as well as maltotriose-modified fourth generation PPI dendrimers with (a) cationic open sugar shell and (b) neutral dense sugar shell that possess a higher biocompatibility, was used. Applying the combination of column chromatography on Cellex P and spectrofluorimetric assays of o-phthaldialdehyde, the final amine condensation products were employed to analyze tissue spermidine and histamine outside the central nervous system. Furthermore, radioenzymatic assay was used to measure histamine levels in the brain. The obtained results indicate that in some tissues, the endogenous concentrations of histamine and spermidine may be affected by dendrimers depending on their dose and type of dendrimers.

Synthesis, DNA interaction, and cytotoxic activity of a novel proflavine-dithiazolidinone pharmacophore.

PubMed

Janovec, Ladislav; Sabolova, Danica; Kozurkova, Maria; Paulíkova, Helena; Kristian, Pavol; Ungvarský, Jan; Moravcíkova, Erika; Bajdichova, Maria; Podhradský, Dusan; Imrich, Jan

2007-01-01

Five novel proflavine-dithiazolidinone derivatives 4a-4e have been designed and synthesized by the reaction of dialkyl acridin-3,6-diyl dithioureas 3a-3e with methyl bromoacetate. The binding affinity of dithiazolidinone hydrochlorides 5a-5e with calf thymus DNA and plasmid (pUC19) DNA was investigated by a variety of spectroscopic techniques including UV-vis, fluorescence, and CD spectroscopy. The effects of 5a-5e on the thermal denaturation profiles of calf thymus DNA were also studied. From spectrophotometric and spectrofluorimetric titrations, the binding constants for the pUC19 DNA-drug complexes were determined (K = 6.2-2.2 x 104 M-1). In vitro cytotoxic activities of compounds 5a-5e toward murine leukemia cell line L1210 and human uterus carcinoma HeLa cells were also examined. 2',2' '-[(Acridin-3,6-diyl)diimino]-3',3' '-dipropyl-1,3-dithiazolidin-4-one hydrochloride (5b) showed the highest activity against these cells with IC50 values of 6.3 microM and 12.9 microM over the course of 72 h.

Characterization of the interactions of the pneumolysoid, Δ6 PLY, with human neutrophils in vitro.

PubMed

Cockeran, R; Steel, H C; Theron, A J; Mitchell, T J; Feldman, C; Anderson, R

2011-11-08

The pneumolysin toxoid, Δ6 PLY, is a prototype pneumococcal protein vaccine candidate. However, its potentially detrimental residual pro-inflammatory interactions with human neutrophils are unknown. In the current study the effects of the toxoid (8-1000 ng/ml) have been compared with those of wild-type pneumolysin (WT/PLY, 8 ng/ml) on neutrophil cytosolic Ca(2+) fluxes, generation of leukotriene B(4) (LTB(4)), and release of matrix metalloproteinase-9 (MMP-9), using spectrofluorimetric, and ELISA procedures (LTB(4) and MMP-9) respectively. Exposure of neutrophils to WT/PLY resulted in influx of Ca(2+) and significant (P<0.05) release of MMP-9 and generation of LTB(4). However, treatment of the cells with Δ6 PLY at concentrations of up to 1000 ng/ml had only trivial effects on Ca(2+) influx and no effects on either release of MMP-9 or LTB(4) production. The observed absence of pro-inflammatory interactions of Δ6 PLY with neutrophils is clearly an important property of this pneumococcal protein vaccine candidate. Copyright © 2011 Elsevier Ltd. All rights reserved.

Non-canonical amino acids bearing thiophene and bithiophene: synthesis by an Ugi multicomponent reaction and studies on ion recognition ability.

PubMed

Esteves, Cátia I C; Raposo, M Manuela M; Costa, Susana P G

2017-05-01

Novel thienyl and bithienyl amino acids with different substituents were obtained by a multicomponent Ugi reaction between a heterocyclic aldehyde, an amine, an acid and an isocyanide. Due to the presence of the sulphur heterocycle at the side chain, these unnatural amino acids are highly emissive and bear extra electron donating atoms so they were tested for their ability to act as fluorescent probes and chemosensors in the recognition of biomedically relevant ions in acetonitrile and acetonitrile/water solutions. The results obtained from spectrophotometric/spectrofluorimetric titrations in the presence of organic and inorganic anions, and alkaline; alkaline-earth and transition metal cations indicated that the bithienyl amino acid bearing a methoxy group is a selective colorimetric chemosensor for Cu 2+ , while the other (bi)thienyl amino acids act as fluorimetric chemosensors with high sensitivity towards Fe 3+ and Cu 2+ in a metal-ligand complex with 1:2 stoichiometry. The photophysical and ion sensing properties of these amino acids confirm their potential as fluorescent probes suitable for incorporation into peptidic frameworks with chemosensory ability.

Sequential determination of nickel and cadmium in tobacco, molasses and refill solutions for e-cigarettes samples by molecular fluorescence.

PubMed

Talio, María Carolina; Alesso, Magdalena; Acosta, Mariano; Wills, Verónica S; Fernández, Liliana P

2017-11-01

In this work, a new procedure was developed for separation and preconcentration of nickel(II) and cadmium(II) in several and varied tobacco samples. Tobacco samples were selected considering the main products consumed by segments of the population, in particular the age (youth) and lifestyle of the consumer. To guarantee representative samples, a randomized strategy of sampling was used. In the first step, a chemofiltration on nylon membrane is carried out employing eosin (Eo) and carbon nanotubes dispersed in sodium dodecylsulfate (SDS) solution (phosphate buffer pH 7). In this condition, Ni(II) was selectively retained on the solid support. After that, the filtrate liquid with Cd(II) was re-conditioned with acetic acid /acetate buffer solution (pH 5) and followed by detection. A spectrofluorimetric determination of both metals was carried out, on the solid support and the filtered aqueous solution, for Ni(II) and Cd(II), respectively. The solid surface fluorescence (SSF) determination was performed at λ em = 545nm (λ ex = 515nm) for Ni(II)-Eo complex and the fluorescence of Cd(II)-Eo was quantified in aqueous solution using λ em = 565nm (λ ex = 540nm). The calibration graphs resulted linear in a range of 0.058-29.35μgL -1 for Ni(II) and 0.124-56.20μgL -1 for Cd(II), with detection limits of 0.019 and 0.041μgL -1 (S/N = 3). The developed methodology shows good sensitivity and adequate selectivity, and it was successfully applied to the determination of trace amounts of nickel and cadmium present in tobacco samples (refill solutions for e-cigarettes, snuff used in narguille (molasses) and traditional tobacco) with satisfactory results. The new methodology was validated by ICP-MS with adequate agreement. The proposed methodology represents a novel fluorescence application to Ni(II) and Cd(II) quantification with sensitivity and accuracy similar to atomic spectroscopies, introducing for the first time the quenching effect on SSF. Copyright © 2017 Elsevier B.V. All rights reserved.

How to test validity in orthodontic research: a mixed dentition analysis example.

PubMed

Donatelli, Richard E; Lee, Shin-Jae

2015-02-01

The data used to test the validity of a prediction method should be different from the data used to generate the prediction model. In this study, we explored whether an independent data set is mandatory for testing the validity of a new prediction method and how validity can be tested without independent new data. Several validation methods were compared in an example using the data from a mixed dentition analysis with a regression model. The validation errors of real mixed dentition analysis data and simulation data were analyzed for increasingly large data sets. The validation results of both the real and the simulation studies demonstrated that the leave-1-out cross-validation method had the smallest errors. The largest errors occurred in the traditional simple validation method. The differences between the validation methods diminished as the sample size increased. The leave-1-out cross-validation method seems to be an optimal validation method for improving the prediction accuracy in a data set with limited sample sizes. Copyright © 2015 American Association of Orthodontists. Published by Elsevier Inc. All rights reserved.

Alteration in plasma corticosterone levels following long term oral administration of lead produces depression like symptoms in rats.

PubMed

Haider, Saida; Saleem, Sadia; Tabassum, Saiqa; Khaliq, Saima; Shamim, Saima; Batool, Zehra; Parveen, Tahira; Inam, Qurat-ul-ain; Haleem, Darakhshan J

2013-03-01

Lead toxicity is known to induce a broad range of physiological, biochemical and behavioral dysfunctions that may result in adverse effects on several organs, including the central nervous system. Long-term exposure to low levels of lead (Pb(2+)) has been shown to produce behavioral deficits in rodents and humans by affecting hypothalamic-pituitary-adrenal (HPA) axis. These deficits are thought to be associated with altered brain monoamine neurotransmission and due to changes in glucocorticoids levels. This study was designed to investigate the effects of Pb(2+)exposure on growth rate, locomotor activity, anxiety, depression, plasma corticosterone and brain serotonin (5-HT) levels in rats. Rats were exposed to lead in drinking water (500 ppm; lead acetate) for 5 weeks. The assessment of depression was done using the forced swimming test (FST). Estimation of brain 5-HT was determined by high-performance liquid chromatography with electrochemical detection. Plasma corticosterone was determined by spectrofluorimetric method. The present study showed that long term exposure to Pb(2+) significantly decreased the food intake followed by the decrease in growth rate in Pb(2+)exposed rats as compared to control group. No significant changes in open field activity were observed following Pb(2+)exposure while significant increase in anxiogenic effect was observed. Increased plasma corticosterone and decreased 5-HT levels were exhibited by Pb(2+)exposed rats as compared to controls. A significant increase in depressive like symptoms was exhibited by Pb(2+)exposed rats as compared to control rats. The results are discussed in the context of Pb(2+) inducing a stress-like response in rats leading to changes in plasma corticosterone and brain 5-HT levels via altering tryptophan pyrrolase activity.

Validation of alternative methods for toxicity testing.

PubMed Central

Bruner, L H; Carr, G J; Curren, R D; Chamberlain, M

1998-01-01

Before nonanimal toxicity tests may be officially accepted by regulatory agencies, it is generally agreed that the validity of the new methods must be demonstrated in an independent, scientifically sound validation program. Validation has been defined as the demonstration of the reliability and relevance of a test method for a particular purpose. This paper provides a brief review of the development of the theoretical aspects of the validation process and updates current thinking about objectively testing the performance of an alternative method in a validation study. Validation of alternative methods for eye irritation testing is a specific example illustrating important concepts. Although discussion focuses on the validation of alternative methods intended to replace current in vivo toxicity tests, the procedures can be used to assess the performance of alternative methods intended for other uses. Images Figure 1 PMID:9599695

Analytical difficulties facing today's regulatory laboratories: issues in method validation.

PubMed

MacNeil, James D

2012-08-01

The challenges facing analytical laboratories today are not unlike those faced in the past, although both the degree of complexity and the rate of change have increased. Challenges such as development and maintenance of expertise, maintenance and up-dating of equipment, and the introduction of new test methods have always been familiar themes for analytical laboratories, but international guidelines for laboratories involved in the import and export testing of food require management of such changes in a context which includes quality assurance, accreditation, and method validation considerations. Decisions as to when a change in a method requires re-validation of the method or on the design of a validation scheme for a complex multi-residue method require a well-considered strategy, based on a current knowledge of international guidance documents and regulatory requirements, as well the laboratory's quality system requirements. Validation demonstrates that a method is 'fit for purpose', so the requirement for validation should be assessed in terms of the intended use of a method and, in the case of change or modification of a method, whether that change or modification may affect a previously validated performance characteristic. In general, method validation involves method scope, calibration-related parameters, method precision, and recovery. Any method change which may affect method scope or any performance parameters will require re-validation. Some typical situations involving change in methods are discussed and a decision process proposed for selection of appropriate validation measures. © 2012 John Wiley & Sons, Ltd.

Cloning and characterization of a riboflavin-binding hexamerin from the larval fat body of a lepidopteran stored grain pest, Corcyra cephalonica.

PubMed

Rao, V Venkat; Ningshen, Thuirei Jacob; Chaitanya, R K; Senthilkumaran, B; Dutta-Gupta, Aparna

2016-01-01

In the present study, a riboflavin-binding hexamerin (RbHex) was cloned and characterized from the larval fat body of Corcyra cephalonica. The complete cDNA (2121bp) encodes a 706-amino acid protein with a molecular mass ~82kDa. Expression of RbHex 82 was predominant in fat body among larval tissues. Further, it is prominently expressed during the last instar larval development. Homology modeling and docking studies predicted riboflavin binding site of the hexamerin. Spectrofluorimetric analysis further confirmed riboflavin release from the hexamerin fraction. Quantitative RT-PCR studies demonstrated hormonal regulation of RbHex 82. 20-Hydroxyecdysone (20HE) had a stimulatory effect on its transcription whereas JH alone did not show any effect. However, JH in the presence of 20HE maintains the RbHex 82 expression which indicates the JH's role as a status quo factor. This study is the first to report the characterization of riboflavin-binding hexamerin in a lepidopteran pest. Further, the possibility of RbHex 82 as a pest control target is discussed. Copyright © 2016 Elsevier Inc. All rights reserved.

Neuroprotective effect of curcumin-loaded lactoferrin nano particles against rotenone induced neurotoxicity.

PubMed

Bollimpelli, V Satish; Kumar, Prashant; Kumari, Sonali; Kondapi, Anand K

2016-05-01

Curcumin is known to have neuroprotective role and possess antioxidant, anti-inflammatory activities. Rotenone, a flavonoid induced neurotoxicity in dopaminergic cells is being widely studied in Parkinson's Disease (PD) research. In the present study, curcumin loaded lactoferrin nano particles prepared by sol-oil chemistry were used to protect dopaminergic cell line SK-N-SH against rotenone induced neurotoxicity. These curcumin loaded nano particles were of 43-60 nm diameter size and around 100 nm hydrodynamic size as assessed by transmission electron microscopy, atomic force microscopy and dynamic light scattering analysis respectively. The encapsulation efficiency was 61.3% ± 2.4%. Cellular uptake of curcumin through these nano particles was confirmed by confocal imaging and spectrofluorimetric analysis. The curcumin loaded lactoferrin nanoparticles showed greater intracellular drug uptake, sustained retention and greater neuroprotection than soluble counterpart. Neuroprotective activity was characterized through viability assays and by estimating ROS levels. Furthermore rotenone induced PD like features were characterized by decrease in tyrosine hydroxylase expression and increase in α-synuclein expression. Taken together curcumin loaded lactoferrin nanoparticles could be a promising drug delivery strategy against neurotoxicity in dopaminergic neurons. Copyright © 2016 Elsevier Ltd. All rights reserved.

Bibliometrics for Social Validation.

PubMed

Hicks, Daniel J

2016-01-01

This paper introduces a bibliometric, citation network-based method for assessing the social validation of novel research, and applies this method to the development of high-throughput toxicology research at the US Environmental Protection Agency. Social validation refers to the acceptance of novel research methods by a relevant scientific community; it is formally independent of the technical validation of methods, and is frequently studied in history, philosophy, and social studies of science using qualitative methods. The quantitative methods introduced here find that high-throughput toxicology methods are spread throughout a large and well-connected research community, which suggests high social validation. Further assessment of social validation involving mixed qualitative and quantitative methods are discussed in the conclusion.

Bibliometrics for Social Validation

PubMed Central

2016-01-01

This paper introduces a bibliometric, citation network-based method for assessing the social validation of novel research, and applies this method to the development of high-throughput toxicology research at the US Environmental Protection Agency. Social validation refers to the acceptance of novel research methods by a relevant scientific community; it is formally independent of the technical validation of methods, and is frequently studied in history, philosophy, and social studies of science using qualitative methods. The quantitative methods introduced here find that high-throughput toxicology methods are spread throughout a large and well-connected research community, which suggests high social validation. Further assessment of social validation involving mixed qualitative and quantitative methods are discussed in the conclusion. PMID:28005974

Considerations regarding the validation of chromatographic mass spectrometric methods for the quantification of endogenous substances in forensics.

PubMed

Hess, Cornelius; Sydow, Konrad; Kueting, Theresa; Kraemer, Michael; Maas, Alexandra

2018-02-01

The requirement for correct evaluation of forensic toxicological results in daily routine work and scientific studies is reliable analytical data based on validated methods. Validation of a method gives the analyst tools to estimate the efficacy and reliability of the analytical method. Without validation, data might be contested in court and lead to unjustified legal consequences for a defendant. Therefore, new analytical methods to be used in forensic toxicology require careful method development and validation of the final method. Until now, there are no publications on the validation of chromatographic mass spectrometric methods for the detection of endogenous substances although endogenous analytes can be important in Forensic Toxicology (alcohol consumption marker, congener alcohols, gamma hydroxy butyric acid, human insulin and C-peptide, creatinine, postmortal clinical parameters). For these analytes, conventional validation instructions cannot be followed completely. In this paper, important practical considerations in analytical method validation for endogenous substances will be discussed which may be used as guidance for scientists wishing to develop and validate analytical methods for analytes produced naturally in the human body. Especially the validation parameters calibration model, analytical limits, accuracy (bias and precision) and matrix effects and recovery have to be approached differently. Highest attention should be paid to selectivity experiments. Copyright © 2017 Elsevier B.V. All rights reserved.

[Sensitive determination of Bi3+ by spectrofluorimetry based on graphene oxide-methylene blue system].

PubMed

Zhai, Qiu-ge; Guo, Peng; Zhou, Lin; Liu, Yan-ming

2014-08-01

Graphene oxide was prepared by the modified Hummers method and characterized by field emission scanning electron microscopy. The interaction of graphene with methylene blue was studied by UV absorption, the intensity of two main absorption peaks of methylene blue decreased significantly after the fluorescence was quenched, and the energy transfer didn't occur because the overlap of the absorption spectrum of GO and the emission spectrum of MB is too small. Therefore, the fluorescence quenching of MB and GO was static. When adding a certain amount of Bi3+ in the graphene-methylene blue system, Bi3+ replaces the methylene blue from the graphene-methylene blue complexes because Bi3+ has the smaller volume and is more positively charged. The methylene blue therefore dissociates from the GO-MB complexes, resulting in the recovery of fluorescence of the system. Furthermore, the fluorescence of the system increases with the increase in the amount of Bi3+ due to the enhanced amount of MB in the system. A novel spectrofluorimetric method was therefore developed for the sensitive determination of Bi3+. Some parameters including the concentration of methylene blue, the amount of graphene oxide, the amount of nitric acid and the sequence of reagent adding were optimized to obtain higher sensitivity. The fluorescence of the system was detected at an emission wavelength of 667 nm with excitation at 690 nm. Under the optimized conditions, the concentration of Bi3+ showed good linear relationships with the fluorescence intensity in the range of 0.5-100 micromol x L(-1), with correlation coefficients of r = 0.9955. The limits of detection for Bi3+ was 1.0 x 10(-8) mol x L(-1) (S/N=3). The selectivity of the proposed method was evaluated and the results showed that 1000-fold K+, Ca+, Na+, Mg2+, Cu2+; 100-fold Fe3+, Be2+, SiO2- Al3+, Ni2+, Sb3+, NO3-, Cl-, F-, and 20-fold Pb2+, Hg2+, Cd2+ had negligible interference with the determination of Bi3+. The method has advantages of sensitivity, rapidness and low cost. It was used for the determination of Bi3+ in environmental water samples inclu- ding pond water, rain water and sewage water with recoveries of 93.4%-105.2%.

78 FR 56718 - Draft Guidance for Industry on Bioanalytical Method Validation; Availability

Federal Register 2010, 2011, 2012, 2013, 2014

2013-09-13

...] Draft Guidance for Industry on Bioanalytical Method Validation; Availability AGENCY: Food and Drug... availability of a draft guidance for industry entitled ``Bioanalytical Method Validation.'' The draft guidance is intended to provide recommendations regarding analytical method development and validation for the...

State of the art in the validation of screening methods for the control of antibiotic residues: is there a need for further development?

PubMed

Gaudin, Valérie

2017-09-01

Screening methods are used as a first-line approach to detect the presence of antibiotic residues in food of animal origin. The validation process guarantees that the method is fit-for-purpose, suited to regulatory requirements, and provides evidence of its performance. This article is focused on intra-laboratory validation. The first step in validation is characterisation of performance, and the second step is the validation itself with regard to pre-established criteria. The validation approaches can be absolute (a single method) or relative (comparison of methods), overall (combination of several characteristics in one) or criterion-by-criterion. Various approaches to validation, in the form of regulations, guidelines or standards, are presented and discussed to draw conclusions on their potential application for different residue screening methods, and to determine whether or not they reach the same conclusions. The approach by comparison of methods is not suitable for screening methods for antibiotic residues. The overall approaches, such as probability of detection (POD) and accuracy profile, are increasingly used in other fields of application. They may be of interest for screening methods for antibiotic residues. Finally, the criterion-by-criterion approach (Decision 2002/657/EC and of European guideline for the validation of screening methods), usually applied to the screening methods for antibiotic residues, introduced a major characteristic and an improvement in the validation, i.e. the detection capability (CCβ). In conclusion, screening methods are constantly evolving, thanks to the development of new biosensors or liquid chromatography coupled to tandem-mass spectrometry (LC-MS/MS) methods. There have been clear changes in validation approaches these last 20 years. Continued progress is required and perspectives for future development of guidelines, regulations and standards for validation are presented here.

New clinical validation method for automated sphygmomanometer: a proposal by Japan ISO-WG for sphygmomanometer standard.

PubMed

Shirasaki, Osamu; Asou, Yosuke; Takahashi, Yukio

2007-12-01

Owing to fast or stepwise cuff deflation, or measuring at places other than the upper arm, the clinical accuracy of most recent automated sphygmomanometers (auto-BPMs) cannot be validated by one-arm simultaneous comparison, which would be the only accurate validation method based on auscultation. Two main alternative methods are provided by current standards, that is, two-arm simultaneous comparison (method 1) and one-arm sequential comparison (method 2); however, the accuracy of these validation methods might not be sufficient to compensate for the suspicious accuracy in lateral blood pressure (BP) differences (LD) and/or BP variations (BPV) between the device and reference readings. Thus, the Japan ISO-WG for sphygmomanometer standards has been studying a new method that might improve validation accuracy (method 3). The purpose of this study is to determine the appropriateness of method 3 by comparing immunity to LD and BPV with those of the current validation methods (methods 1 and 2). The validation accuracy of the above three methods was assessed in human participants [N=120, 45+/-15.3 years (mean+/-SD)]. An oscillometric automated monitor, Omron HEM-762, was used as the tested device. When compared with the others, methods 1 and 3 showed a smaller intra-individual standard deviation of device error (SD1), suggesting their higher reproducibility of validation. The SD1 by method 2 (P=0.004) significantly correlated with the participant's BP, supporting our hypothesis that the increased SD of device error by method 2 is at least partially caused by essential BPV. Method 3 showed a significantly (P=0.0044) smaller interparticipant SD of device error (SD2), suggesting its higher interparticipant consistency of validation. Among the methods of validation of the clinical accuracy of auto-BPMs, method 3, which showed the highest reproducibility and highest interparticipant consistency, can be proposed as being the most appropriate.

The Importance of Method Selection in Determining Product Integrity for Nutrition Research1234

PubMed Central

Mudge, Elizabeth M; Brown, Paula N

2016-01-01

The American Herbal Products Association estimates that there as many as 3000 plant species in commerce. The FDA estimates that there are about 85,000 dietary supplement products in the marketplace. The pace of product innovation far exceeds that of analytical methods development and validation, with new ingredients, matrixes, and combinations resulting in an analytical community that has been unable to keep up. This has led to a lack of validated analytical methods for dietary supplements and to inappropriate method selection where methods do exist. Only after rigorous validation procedures to ensure that methods are fit for purpose should they be used in a routine setting to verify product authenticity and quality. By following systematic procedures and establishing performance requirements for analytical methods before method development and validation, methods can be developed that are both valid and fit for purpose. This review summarizes advances in method selection, development, and validation regarding herbal supplement analysis and provides several documented examples of inappropriate method selection and application. PMID:26980823

The Importance of Method Selection in Determining Product Integrity for Nutrition Research.

PubMed

Mudge, Elizabeth M; Betz, Joseph M; Brown, Paula N

2016-03-01

The American Herbal Products Association estimates that there as many as 3000 plant species in commerce. The FDA estimates that there are about 85,000 dietary supplement products in the marketplace. The pace of product innovation far exceeds that of analytical methods development and validation, with new ingredients, matrixes, and combinations resulting in an analytical community that has been unable to keep up. This has led to a lack of validated analytical methods for dietary supplements and to inappropriate method selection where methods do exist. Only after rigorous validation procedures to ensure that methods are fit for purpose should they be used in a routine setting to verify product authenticity and quality. By following systematic procedures and establishing performance requirements for analytical methods before method development and validation, methods can be developed that are both valid and fit for purpose. This review summarizes advances in method selection, development, and validation regarding herbal supplement analysis and provides several documented examples of inappropriate method selection and application. © 2016 American Society for Nutrition.

Method validation for chemical composition determination by electron microprobe with wavelength dispersive spectrometer

NASA Astrophysics Data System (ADS)

Herrera-Basurto, R.; Mercader-Trejo, F.; Muñoz-Madrigal, N.; Juárez-García, J. M.; Rodriguez-López, A.; Manzano-Ramírez, A.

2016-07-01

The main goal of method validation is to demonstrate that the method is suitable for its intended purpose. One of the advantages of analytical method validation is translated into a level of confidence about the measurement results reported to satisfy a specific objective. Elemental composition determination by wavelength dispersive spectrometer (WDS) microanalysis has been used over extremely wide areas, mainly in the field of materials science, impurity determinations in geological, biological and food samples. However, little information is reported about the validation of the applied methods. Herein, results of the in-house method validation for elemental composition determination by WDS are shown. SRM 482, a binary alloy Cu-Au of different compositions, was used during the validation protocol following the recommendations for method validation proposed by Eurachem. This paper can be taken as a reference for the evaluation of the validation parameters more frequently requested to get the accreditation under the requirements of the ISO/IEC 17025 standard: selectivity, limit of detection, linear interval, sensitivity, precision, trueness and uncertainty. A model for uncertainty estimation was proposed including systematic and random errors. In addition, parameters evaluated during the validation process were also considered as part of the uncertainty model.

Meeting Report: Validation of Toxicogenomics-Based Test Systems: ECVAM–ICCVAM/NICEATM Considerations for Regulatory Use

PubMed Central

Corvi, Raffaella; Ahr, Hans-Jürgen; Albertini, Silvio; Blakey, David H.; Clerici, Libero; Coecke, Sandra; Douglas, George R.; Gribaldo, Laura; Groten, John P.; Haase, Bernd; Hamernik, Karen; Hartung, Thomas; Inoue, Tohru; Indans, Ian; Maurici, Daniela; Orphanides, George; Rembges, Diana; Sansone, Susanna-Assunta; Snape, Jason R.; Toda, Eisaku; Tong, Weida; van Delft, Joost H.; Weis, Brenda; Schechtman, Leonard M.

2006-01-01

This is the report of the first workshop “Validation of Toxicogenomics-Based Test Systems” held 11–12 December 2003 in Ispra, Italy. The workshop was hosted by the European Centre for the Validation of Alternative Methods (ECVAM) and organized jointly by ECVAM, the U.S. Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM), and the National Toxicology Program (NTP) Interagency Center for the Evaluation of Alternative Toxicological Methods (NICEATM). The primary aim of the workshop was for participants to discuss and define principles applicable to the validation of toxicogenomics platforms as well as validation of specific toxicologic test methods that incorporate toxicogenomics technologies. The workshop was viewed as an opportunity for initiating a dialogue between technologic experts, regulators, and the principal validation bodies and for identifying those factors to which the validation process would be applicable. It was felt that to do so now, as the technology is evolving and associated challenges are identified, would be a basis for the future validation of the technology when it reaches the appropriate stage. Because of the complexity of the issue, different aspects of the validation of toxicogenomics-based test methods were covered. The three focus areas include a) biologic validation of toxicogenomics-based test methods for regulatory decision making, b) technical and bioinformatics aspects related to validation, and c) validation issues as they relate to regulatory acceptance and use of toxicogenomics-based test methods. In this report we summarize the discussions and describe in detail the recommendations for future direction and priorities. PMID:16507466

Meeting report: Validation of toxicogenomics-based test systems: ECVAM-ICCVAM/NICEATM considerations for regulatory use.

PubMed

Corvi, Raffaella; Ahr, Hans-Jürgen; Albertini, Silvio; Blakey, David H; Clerici, Libero; Coecke, Sandra; Douglas, George R; Gribaldo, Laura; Groten, John P; Haase, Bernd; Hamernik, Karen; Hartung, Thomas; Inoue, Tohru; Indans, Ian; Maurici, Daniela; Orphanides, George; Rembges, Diana; Sansone, Susanna-Assunta; Snape, Jason R; Toda, Eisaku; Tong, Weida; van Delft, Joost H; Weis, Brenda; Schechtman, Leonard M

2006-03-01

This is the report of the first workshop "Validation of Toxicogenomics-Based Test Systems" held 11-12 December 2003 in Ispra, Italy. The workshop was hosted by the European Centre for the Validation of Alternative Methods (ECVAM) and organized jointly by ECVAM, the U.S. Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM), and the National Toxicology Program (NTP) Interagency Center for the Evaluation of Alternative Toxicological Methods (NICEATM). The primary aim of the workshop was for participants to discuss and define principles applicable to the validation of toxicogenomics platforms as well as validation of specific toxicologic test methods that incorporate toxicogenomics technologies. The workshop was viewed as an opportunity for initiating a dialogue between technologic experts, regulators, and the principal validation bodies and for identifying those factors to which the validation process would be applicable. It was felt that to do so now, as the technology is evolving and associated challenges are identified, would be a basis for the future validation of the technology when it reaches the appropriate stage. Because of the complexity of the issue, different aspects of the validation of toxicogenomics-based test methods were covered. The three focus areas include a) biologic validation of toxicogenomics-based test methods for regulatory decision making, b) technical and bioinformatics aspects related to validation, and c) validation issues as they relate to regulatory acceptance and use of toxicogenomics-based test methods. In this report we summarize the discussions and describe in detail the recommendations for future direction and priorities.

Validation approach for a fast and simple targeted screening method for 75 antibiotics in meat and aquaculture products using LC-MS/MS.

PubMed

Dubreil, Estelle; Gautier, Sophie; Fourmond, Marie-Pierre; Bessiral, Mélaine; Gaugain, Murielle; Verdon, Eric; Pessel, Dominique

2017-04-01

An approach is described to validate a fast and simple targeted screening method for antibiotic analysis in meat and aquaculture products by LC-MS/MS. The strategy of validation was applied for a panel of 75 antibiotics belonging to different families, i.e., penicillins, cephalosporins, sulfonamides, macrolides, quinolones and phenicols. The samples were extracted once with acetonitrile, concentrated by evaporation and injected into the LC-MS/MS system. The approach chosen for the validation was based on the Community Reference Laboratory (CRL) guidelines for the validation of screening qualitative methods. The aim of the validation was to prove sufficient sensitivity of the method to detect all the targeted antibiotics at the level of interest, generally the maximum residue limit (MRL). A robustness study was also performed to test the influence of different factors. The validation showed that the method is valid to detect and identify 73 antibiotics of the 75 antibiotics studied in meat and aquaculture products at the validation levels.

Experimental Validation of Model Updating and Damage Detection via Eigenvalue Sensitivity Methods with Artificial Boundary Conditions

DTIC Science & Technology

2017-09-01

VALIDATION OF MODEL UPDATING AND DAMAGE DETECTION VIA EIGENVALUE SENSITIVITY METHODS WITH ARTIFICIAL BOUNDARY CONDITIONS by Matthew D. Bouwense...VALIDATION OF MODEL UPDATING AND DAMAGE DETECTION VIA EIGENVALUE SENSITIVITY METHODS WITH ARTIFICIAL BOUNDARY CONDITIONS 5. FUNDING NUMBERS 6. AUTHOR...unlimited. EXPERIMENTAL VALIDATION OF MODEL UPDATING AND DAMAGE DETECTION VIA EIGENVALUE SENSITIVITY METHODS WITH ARTIFICIAL BOUNDARY

Brazilian Center for the Validation of Alternative Methods (BraCVAM) and the process of validation in Brazil.

PubMed

Presgrave, Octavio; Moura, Wlamir; Caldeira, Cristiane; Pereira, Elisabete; Bôas, Maria H Villas; Eskes, Chantra

2016-03-01

The need for the creation of a Brazilian centre for the validation of alternative methods was recognised in 2008, and members of academia, industry and existing international validation centres immediately engaged with the idea. In 2012, co-operation between the Oswaldo Cruz Foundation (FIOCRUZ) and the Brazilian Health Surveillance Agency (ANVISA) instigated the establishment of the Brazilian Center for the Validation of Alternative Methods (BraCVAM), which was officially launched in 2013. The Brazilian validation process follows OECD Guidance Document No. 34, where BraCVAM functions as the focal point to identify and/or receive requests from parties interested in submitting tests for validation. BraCVAM then informs the Brazilian National Network on Alternative Methods (RENaMA) of promising assays, which helps with prioritisation and contributes to the validation studies of selected assays. A Validation Management Group supervises the validation study, and the results obtained are peer-reviewed by an ad hoc Scientific Review Committee, organised under the auspices of BraCVAM. Based on the peer-review outcome, BraCVAM will prepare recommendations on the validated test method, which will be sent to the National Council for the Control of Animal Experimentation (CONCEA). CONCEA is in charge of the regulatory adoption of all validated test methods in Brazil, following an open public consultation. 2016 FRAME.

Identification and quantification of carbamate pesticides in dried lime tree flowers by means of excitation-emission molecular fluorescence and parallel factor analysis when quenching effect exists.

PubMed

Rubio, L; Ortiz, M C; Sarabia, L A

2014-04-11

A non-separative, fast and inexpensive spectrofluorimetric method based on the second order calibration of excitation-emission fluorescence matrices (EEMs) was proposed for the determination of carbaryl, carbendazim and 1-naphthol in dried lime tree flowers. The trilinearity property of three-way data was used to handle the intrinsic fluorescence of lime flowers and the difference in the fluorescence intensity of each analyte. It also made possible to identify unequivocally each analyte. Trilinearity of the data tensor guarantees the uniqueness of the solution obtained through parallel factor analysis (PARAFAC), so the factors of the decomposition match up with the analytes. In addition, an experimental procedure was proposed to identify, with three-way data, the quenching effect produced by the fluorophores of the lime flowers. This procedure also enabled the selection of the adequate dilution of the lime flowers extract to minimize the quenching effect so the three analytes can be quantified. Finally, the analytes were determined using the standard addition method for a calibration whose standards were chosen with a D-optimal design. The three analytes were unequivocally identified by the correlation between the pure spectra and the PARAFAC excitation and emission spectral loadings. The trueness was established by the accuracy line "calculated concentration versus added concentration" in all cases. Better decision limit values (CCα), in x0=0 with the probability of false positive fixed at 0.05, were obtained for the calibration performed in pure solvent: 2.97 μg L(-1) for 1-naphthol, 3.74 μg L(-1) for carbaryl and 23.25 μg L(-1) for carbendazim. The CCα values for the second calibration carried out in matrix were 1.61, 4.34 and 51.75 μg L(-1) respectively; while the values obtained considering only the pure samples as calibration set were: 2.65, 8.61 and 28.7 μg L(-1), respectively. Copyright © 2014 Elsevier B.V. All rights reserved.

Likelihood ratio data to report the validation of a forensic fingerprint evaluation method.

PubMed

Ramos, Daniel; Haraksim, Rudolf; Meuwly, Didier

2017-02-01

Data to which the authors refer to throughout this article are likelihood ratios (LR) computed from the comparison of 5-12 minutiae fingermarks with fingerprints. These LRs data are used for the validation of a likelihood ratio (LR) method in forensic evidence evaluation. These data present a necessary asset for conducting validation experiments when validating LR methods used in forensic evidence evaluation and set up validation reports. These data can be also used as a baseline for comparing the fingermark evidence in the same minutiae configuration as presented in (D. Meuwly, D. Ramos, R. Haraksim,) [1], although the reader should keep in mind that different feature extraction algorithms and different AFIS systems used may produce different LRs values. Moreover, these data may serve as a reproducibility exercise, in order to train the generation of validation reports of forensic methods, according to [1]. Alongside the data, a justification and motivation for the use of methods is given. These methods calculate LRs from the fingerprint/mark data and are subject to a validation procedure. The choice of using real forensic fingerprint in the validation and simulated data in the development is described and justified. Validation criteria are set for the purpose of validation of the LR methods, which are used to calculate the LR values from the data and the validation report. For privacy and data protection reasons, the original fingerprint/mark images cannot be shared. But these images do not constitute the core data for the validation, contrarily to the LRs that are shared.

Valid methods: the quality assurance of test method development, validation, approval, and transfer for veterinary testing laboratories.

PubMed

Wiegers, Ann L

2003-07-01

Third-party accreditation is a valuable tool to demonstrate a laboratory's competence to conduct testing. Accreditation, internationally and in the United States, has been discussed previously. However, accreditation is only I part of establishing data credibility. A validated test method is the first component of a valid measurement system. Validation is defined as confirmation by examination and the provision of objective evidence that the particular requirements for a specific intended use are fulfilled. The international and national standard ISO/IEC 17025 recognizes the importance of validated methods and requires that laboratory-developed methods or methods adopted by the laboratory be appropriate for the intended use. Validated methods are therefore required and their use agreed to by the client (i.e., end users of the test results such as veterinarians, animal health programs, and owners). ISO/IEC 17025 also requires that the introduction of methods developed by the laboratory for its own use be a planned activity conducted by qualified personnel with adequate resources. This article discusses considerations and recommendations for the conduct of veterinary diagnostic test method development, validation, evaluation, approval, and transfer to the user laboratory in the ISO/IEC 17025 environment. These recommendations are based on those of nationally and internationally accepted standards and guidelines, as well as those of reputable and experienced technical bodies. They are also based on the author's experience in the evaluation of method development and transfer projects, validation data, and the implementation of quality management systems in the area of method development.

Validity and reliability of bioelectrical impedance analysis and skinfold thickness in predicting body fat in military personnel.

PubMed

Aandstad, Anders; Holtberget, Kristian; Hageberg, Rune; Holme, Ingar; Anderssen, Sigmund A

2014-02-01

Previous studies show that body composition is related to injury risk and physical performance in soldiers. Thus, valid methods for measuring body composition in military personnel are needed. The frequently used body mass index method is not a valid measure of body composition in soldiers, but reliability and validity of alternative field methods are less investigated in military personnel. Thus, we carried out test and retest of skinfold (SKF), single frequency bioelectrical impedance analysis (SF-BIA), and multifrequency bioelectrical impedance analysis measurements in 65 male and female soldiers. Several validated equations were used to predict percent body fat from these methods. Dual-energy X-ray absorptiometry was also measured, and acted as the criterion method. Results showed that SF-BIA was the most reliable method in both genders. In women, SF-BIA was also the most valid method, whereas SKF or a combination of SKF and SF-BIA produced the highest validity in men. Reliability and validity varied substantially among the equations examined. The best methods and equations produced test-retest 95% limits of agreement below ±1% points, whereas the corresponding validity figures were ±3.5% points. Each investigator and practitioner must consider whether such measurement errors are acceptable for its specific use. Reprint & Copyright © 2014 Association of Military Surgeons of the U.S.

Construct Validity and Scoring Methods of the World Health Organization: Health and Work Performance Questionnaire Among Workers With Arthritis and Rheumatological Conditions.

PubMed

AlHeresh, Rawan; LaValley, Michael P; Coster, Wendy; Keysor, Julie J

2017-06-01

To evaluate construct validity and scoring methods of the world health organization-health and work performance questionnaire (HPQ) for people with arthritis. Construct validity was examined through hypothesis testing using the recommended guidelines of the consensus-based standards for the selection of health measurement instruments (COSMIN). The HPQ using the absolute scoring method showed moderate construct validity as four of the seven hypotheses were met. The HPQ using the relative scoring method had weak construct validity as only one of the seven hypotheses were met. The absolute scoring method for the HPQ is superior in construct validity to the relative scoring method in assessing work performance among people with arthritis and related rheumatic conditions; however, more research is needed to further explore other psychometric properties of the HPQ.

Comparative assessment of bioanalytical method validation guidelines for pharmaceutical industry.

PubMed

Kadian, Naveen; Raju, Kanumuri Siva Rama; Rashid, Mamunur; Malik, Mohd Yaseen; Taneja, Isha; Wahajuddin, Muhammad

2016-07-15

The concepts, importance, and application of bioanalytical method validation have been discussed for a long time and validation of bioanalytical methods is widely accepted as pivotal before they are taken into routine use. United States Food and Drug Administration (USFDA) guidelines issued in 2001 have been referred for every guideline released ever since; may it be European Medical Agency (EMA) Europe, National Health Surveillance Agency (ANVISA) Brazil, Ministry of Health and Labour Welfare (MHLW) Japan or any other guideline in reference to bioanalytical method validation. After 12 years, USFDA released its new draft guideline for comments in 2013, which covers the latest parameters or topics encountered in bioanalytical method validation and approached towards the harmonization of bioanalytical method validation across the globe. Even though the regulatory agencies have general agreement, significant variations exist in acceptance criteria and methodology. The present review highlights the variations, similarities and comparison between bioanalytical method validation guidelines issued by major regulatory authorities worldwide. Additionally, other evaluation parameters such as matrix effect, incurred sample reanalysis including other stability aspects have been discussed to provide an ease of access for designing a bioanalytical method and its validation complying with the majority of drug authority guidelines. Copyright © 2016. Published by Elsevier B.V.

Fuzzy-logic based strategy for validation of multiplex methods: example with qualitative GMO assays.

PubMed

Bellocchi, Gianni; Bertholet, Vincent; Hamels, Sandrine; Moens, W; Remacle, José; Van den Eede, Guy

2010-02-01

This paper illustrates the advantages that a fuzzy-based aggregation method could bring into the validation of a multiplex method for GMO detection (DualChip GMO kit, Eppendorf). Guidelines for validation of chemical, bio-chemical, pharmaceutical and genetic methods have been developed and ad hoc validation statistics are available and routinely used, for in-house and inter-laboratory testing, and decision-making. Fuzzy logic allows summarising the information obtained by independent validation statistics into one synthetic indicator of overall method performance. The microarray technology, introduced for simultaneous identification of multiple GMOs, poses specific validation issues (patterns of performance for a variety of GMOs at different concentrations). A fuzzy-based indicator for overall evaluation is illustrated in this paper, and applied to validation data for different genetically modified elements. Remarks were drawn on the analytical results. The fuzzy-logic based rules were shown to be applicable to improve interpretation of results and facilitate overall evaluation of the multiplex method.

76 FR 28664 - Method 301-Field Validation of Pollutant Measurement Methods From Various Waste Media

Federal Register 2010, 2011, 2012, 2013, 2014

2011-05-18

... ENVIRONMENTAL PROTECTION AGENCY 40 CFR Part 63 [OAR-2004-0080, FRL-9306-8] RIN 2060-AF00 Method 301--Field Validation of Pollutant Measurement Methods From Various Waste Media AGENCY: Environmental Protection Agency (EPA). ACTION: Final rule. SUMMARY: This action amends EPA's Method 301, Field Validation...

Validation of Alternative In Vitro Methods to Animal Testing: Concepts, Challenges, Processes and Tools.

PubMed

Griesinger, Claudius; Desprez, Bertrand; Coecke, Sandra; Casey, Warren; Zuang, Valérie

This chapter explores the concepts, processes, tools and challenges relating to the validation of alternative methods for toxicity and safety testing. In general terms, validation is the process of assessing the appropriateness and usefulness of a tool for its intended purpose. Validation is routinely used in various contexts in science, technology, the manufacturing and services sectors. It serves to assess the fitness-for-purpose of devices, systems, software up to entire methodologies. In the area of toxicity testing, validation plays an indispensable role: "alternative approaches" are increasingly replacing animal models as predictive tools and it needs to be demonstrated that these novel methods are fit for purpose. Alternative approaches include in vitro test methods, non-testing approaches such as predictive computer models up to entire testing and assessment strategies composed of method suites, data sources and decision-aiding tools. Data generated with alternative approaches are ultimately used for decision-making on public health and the protection of the environment. It is therefore essential that the underlying methods and methodologies are thoroughly characterised, assessed and transparently documented through validation studies involving impartial actors. Importantly, validation serves as a filter to ensure that only test methods able to produce data that help to address legislative requirements (e.g. EU's REACH legislation) are accepted as official testing tools and, owing to the globalisation of markets, recognised on international level (e.g. through inclusion in OECD test guidelines). Since validation creates a credible and transparent evidence base on test methods, it provides a quality stamp, supporting companies developing and marketing alternative methods and creating considerable business opportunities. Validation of alternative methods is conducted through scientific studies assessing two key hypotheses, reliability and relevance of the test method for a given purpose. Relevance encapsulates the scientific basis of the test method, its capacity to predict adverse effects in the "target system" (i.e. human health or the environment) as well as its applicability for the intended purpose. In this chapter we focus on the validation of non-animal in vitro alternative testing methods and review the concepts, challenges, processes and tools fundamental to the validation of in vitro methods intended for hazard testing of chemicals. We explore major challenges and peculiarities of validation in this area. Based on the notion that validation per se is a scientific endeavour that needs to adhere to key scientific principles, namely objectivity and appropriate choice of methodology, we examine basic aspects of study design and management, and provide illustrations of statistical approaches to describe predictive performance of validated test methods as well as their reliability.

Software validation applied to spreadsheets used in laboratories working under ISO/IEC 17025

NASA Astrophysics Data System (ADS)

Banegas, J. M.; Orué, M. W.

2016-07-01

Several documents deal with software validation. Nevertheless, more are too complex to be applied to validate spreadsheets - surely the most used software in laboratories working under ISO/IEC 17025. The method proposed in this work is intended to be directly applied to validate spreadsheets. It includes a systematic way to document requirements, operational aspects regarding to validation, and a simple method to keep records of validation results and modifications history. This method is actually being used in an accredited calibration laboratory, showing to be practical and efficient.

Comparative assessment of three standardized robotic surgery training methods.

PubMed

Hung, Andrew J; Jayaratna, Isuru S; Teruya, Kara; Desai, Mihir M; Gill, Inderbir S; Goh, Alvin C

2013-10-01

To evaluate three standardized robotic surgery training methods, inanimate, virtual reality and in vivo, for their construct validity. To explore the concept of cross-method validity, where the relative performance of each method is compared. Robotic surgical skills were prospectively assessed in 49 participating surgeons who were classified as follows: 'novice/trainee': urology residents, previous experience <30 cases (n = 38) and 'experts': faculty surgeons, previous experience ≥30 cases (n = 11). Three standardized, validated training methods were used: (i) structured inanimate tasks; (ii) virtual reality exercises on the da Vinci Skills Simulator (Intuitive Surgical, Sunnyvale, CA, USA); and (iii) a standardized robotic surgical task in a live porcine model with performance graded by the Global Evaluative Assessment of Robotic Skills (GEARS) tool. A Kruskal-Wallis test was used to evaluate performance differences between novices and experts (construct validity). Spearman's correlation coefficient (ρ) was used to measure the association of performance across inanimate, simulation and in vivo methods (cross-method validity). Novice and expert surgeons had previously performed a median (range) of 0 (0-20) and 300 (30-2000) robotic cases, respectively (P < 0.001). Construct validity: experts consistently outperformed residents with all three methods (P < 0.001). Cross-method validity: overall performance of inanimate tasks significantly correlated with virtual reality robotic performance (ρ = -0.7, P < 0.001) and in vivo robotic performance based on GEARS (ρ = -0.8, P < 0.0001). Virtual reality performance and in vivo tissue performance were also found to be strongly correlated (ρ = 0.6, P < 0.001). We propose the novel concept of cross-method validity, which may provide a method of evaluating the relative value of various forms of skills education and assessment. We externally confirmed the construct validity of each featured training tool. © 2013 BJU International.

Isolation and N-terminal sequencing of a novel cadmium-binding protein from Boletus edulis

NASA Astrophysics Data System (ADS)

Collin-Hansen, C.; Andersen, R. A.; Steinnes, E.

2003-05-01

A Cd-binding protein was isolated from the popular edible mushroom Boletus edulis, which is a hyperaccumulator of both Cd and Hg. Wild-growing samples of B. edulis were collected from soils rich in Cd. Cd radiotracer was added to the crude protein preparation obtained from ethanol precipitation of heat-treated cytosol. Proteins were then further separated in two consecutive steps; gel filtration and anion exchange chromatography. In both steps the Cd radiotracer profile showed only one distinct peak, which corresponded well with the profiles of endogenous Cd obtained by atomic absorption spectrophotometry (AAS). Concentrations of the essential elements Cu and Zn were low in the protein fractions high in Cd. N-terminal sequencing performed on the Cd-binding protein fractions revealed a protein with a novel amino acid sequence, which contained aromatic amino acids as well as proline. Both the N-terminal sequencing and spectrofluorimetric analysis with EDTA and ABD-F (4-aminosulfonyl-7-fluoro-2, 1, 3-benzoxadiazole) failed to detect cysteine in the Cd-binding fractions. These findings conclude that the novel protein does not belong to the metallothionein family. The results suggest a role for the protein in Cd transport and storage, and they are of importance in view of toxicology and food chemistry, but also for environmental protection.

Supramolecular polymeric chemosensor for biomedical applications: design and synthesis of a luminescent zinc metallopolymer as a chemosensor for adenine detection.

PubMed

Chow, Cheuk-Fai

2012-11-01

Adenine is an important bio-molecule that plays many crucial roles in food safety and biomedical diagnostics. Differentiating adenine from a mixture of adenosine and other nucleic bases (guanine, thymine, cytosine, and uracil) is particularly important for both biological and clinical applications. A neutral Zn(II) metallosupramolecular polymer based on acyl hydrazone derived coordination centres (P1) were generated through self-assembly polymerization. It is a linear coordination polymer that behaves like self-standing film. The synthesis, (1)H-NMR characterization, and spectroscopic properties of this supramolecular material are reported. P1 was found to be a chemosensor specific to adenine, with a luminescent enhancement. The binding properties of P1 with common nucleic bases and nucleosides reveal that this supramolecular polymer is very selective to adenine molecules (~20 to 420 times more selectivity than other nucleic bases). The formation constant (K) of P1 to adenine was found to be log K = 4.10 ± 0.02. This polymeric chemosensor produces a specific response to adenine down to 90 ppb. Spectrofluorimetric and (1)H-NMR titration studies showed that the P1 polymer allows each Zn(II) coordination centre to bind to two adenine molecules through hydrogen bonding with their imine and hydrazone protons.

Design and synthesis of heterobimetallic Ru(II)-Ln(III) complexes as chemodosimetric ensembles for the detection of biogenic amine odorants.

PubMed

Chow, Cheuk-Fai; Lam, Michael H W; Wong, Wai-Yeung

2013-09-03

The detection of neutral biogenic amines plays a crucial role in food safety. Three new heterobimetallic Ru(II)-Ln(III) donor-acceptor complexes, KPrRu, KNdRu, and KSmRu, K{[Ru((II))((t)Bubpy)(CN)4]2-Ln((III))(H2O)4} (where (t)Bubpy = 4,4'-di-tert-butyl-2,2'-bipyridine), have been synthesized and characterized. Their photophysical and X-ray crystallographic data were reported in this study. These complexes were found to be selective for biogenic amine vapors, such as histamine, putrescine, and spermidine, with a detection limit down to the ppb level. The sensitivities of these complexes to the amines were recorded as ~log K = 3.6-5.0. Submicron rods of the complexes, with a nanoscale diameter and microscale length, were obtained through a simple precipitation process. Free-standing polymeric films with different degrees of porosity were fabricated by blending the submicron rods with polystyrene polymer. The polymer with the highest level of porosity exhibited the strongest luminescence enhancement after amine exposure. Real time monitoring of gaseous biogenic amines was applied to real fish samples (Atlantic mackerel) by studying the spectrofluorimetric responses of the Ru(II)-Ln(III) blended polymer film.

Standard Setting Methods for Pass/Fail Decisions on High-Stakes Objective Structured Clinical Examinations: A Validity Study.

PubMed

Yousuf, Naveed; Violato, Claudio; Zuberi, Rukhsana W

2015-01-01

CONSTRUCT: Authentic standard setting methods will demonstrate high convergent validity evidence of their outcomes, that is, cutoff scores and pass/fail decisions, with most other methods when compared with each other. The objective structured clinical examination (OSCE) was established for valid, reliable, and objective assessment of clinical skills in health professions education. Various standard setting methods have been proposed to identify objective, reliable, and valid cutoff scores on OSCEs. These methods may identify different cutoff scores for the same examinations. Identification of valid and reliable cutoff scores for OSCEs remains an important issue and a challenge. Thirty OSCE stations administered at least twice in the years 2010-2012 to 393 medical students in Years 2 and 3 at Aga Khan University are included. Psychometric properties of the scores are determined. Cutoff scores and pass/fail decisions of Wijnen, Cohen, Mean-1.5SD, Mean-1SD, Angoff, borderline group and borderline regression (BL-R) methods are compared with each other and with three variants of cluster analysis using repeated measures analysis of variance and Cohen's kappa. The mean psychometric indices on the 30 OSCE stations are reliability coefficient = 0.76 (SD = 0.12); standard error of measurement = 5.66 (SD = 1.38); coefficient of determination = 0.47 (SD = 0.19), and intergrade discrimination = 7.19 (SD = 1.89). BL-R and Wijnen methods show the highest convergent validity evidence among other methods on the defined criteria. Angoff and Mean-1.5SD demonstrated least convergent validity evidence. The three cluster variants showed substantial convergent validity with borderline methods. Although there was a high level of convergent validity of Wijnen method, it lacks the theoretical strength to be used for competency-based assessments. The BL-R method is found to show the highest convergent validity evidences for OSCEs with other standard setting methods used in the present study. We also found that cluster analysis using mean method can be used for quality assurance of borderline methods. These findings should be further confirmed by studies in other settings.

An Engineering Method of Civil Jet Requirements Validation Based on Requirements Project Principle

NASA Astrophysics Data System (ADS)

Wang, Yue; Gao, Dan; Mao, Xuming

2018-03-01

A method of requirements validation is developed and defined to meet the needs of civil jet requirements validation in product development. Based on requirements project principle, this method will not affect the conventional design elements, and can effectively connect the requirements with design. It realizes the modern civil jet development concept, which is “requirement is the origin, design is the basis”. So far, the method has been successfully applied in civil jet aircraft development in China. Taking takeoff field length as an example, the validation process and the validation method of the requirements are detailed introduced in the study, with the hope of providing the experiences to other civil jet product design.

Validity and Feasibility of a Digital Diet Estimation Method for Use with Preschool Children: A Pilot Study

ERIC Educational Resources Information Center

Nicklas, Theresa A.; O'Neil, Carol E.; Stuff, Janice; Goodell, Lora Suzanne; Liu, Yan; Martin, Corby K.

2012-01-01

Objective: The goal of the study was to assess the validity and feasibility of a digital diet estimation method for use with preschool children in "Head Start." Methods: Preschool children and their caregivers participated in validation (n = 22) and feasibility (n = 24) pilot studies. Validity was determined in the metabolic research unit using…

VALIDATION OF ANALYTICAL METHODS AND INSTRUMENTATION FOR BERYLLIUM MEASUREMENT: REVIEW AND SUMMARY OF AVAILABLE GUIDES, PROCEDURES, AND PROTOCOLS

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ekechukwu, A.

This document proposes to provide a listing of available sources which can be used to validate analytical methods and/or instrumentation for beryllium determination. A literature review was conducted of available standard methods and publications used for method validation and/or quality control. A comprehensive listing of the articles, papers, and books reviewed is given in Appendix 1. Available validation documents and guides are listed in the appendix; each has a brief description of application and use. In the referenced sources, there are varying approaches to validation and varying descriptions of validation at different stages in method development. This discussion focuses onmore » validation and verification of fully developed methods and instrumentation that have been offered up for use or approval by other laboratories or official consensus bodies such as ASTM International, the International Standards Organization (ISO) and the Association of Official Analytical Chemists (AOAC). This review was conducted as part of a collaborative effort to investigate and improve the state of validation for measuring beryllium in the workplace and the environment. Documents and publications from the United States and Europe are included. Unless otherwise specified, all documents were published in English.« less

Validation in Support of Internationally Harmonised OECD Test Guidelines for Assessing the Safety of Chemicals.

PubMed

Gourmelon, Anne; Delrue, Nathalie

Ten years elapsed since the OECD published the Guidance document on the validation and international regulatory acceptance of test methods for hazard assessment. Much experience has been gained since then in validation centres, in countries and at the OECD on a variety of test methods that were subjected to validation studies. This chapter reviews validation principles and highlights common features that appear to be important for further regulatory acceptance across studies. Existing OECD-agreed validation principles will most likely generally remain relevant and applicable to address challenges associated with the validation of future test methods. Some adaptations may be needed to take into account the level of technique introduced in test systems, but demonstration of relevance and reliability will continue to play a central role as pre-requisite for the regulatory acceptance. Demonstration of relevance will become more challenging for test methods that form part of a set of predictive tools and methods, and that do not stand alone. OECD is keen on ensuring that while these concepts evolve, countries can continue to rely on valid methods and harmonised approaches for an efficient testing and assessment of chemicals.

Toward a Unified Validation Framework in Mixed Methods Research

ERIC Educational Resources Information Center

Dellinger, Amy B.; Leech, Nancy L.

2007-01-01

The primary purpose of this article is to further discussions of validity in mixed methods research by introducing a validation framework to guide thinking about validity in this area. To justify the use of this framework, the authors discuss traditional terminology and validity criteria for quantitative and qualitative research, as well as…

A systematic and critical review on bioanalytical method validation using the example of simultaneous quantitation of antidiabetic agents in blood.

PubMed

Fachi, Mariana Millan; Leonart, Letícia Paula; Cerqueira, Letícia Bonancio; Pontes, Flavia Lada Degaut; de Campos, Michel Leandro; Pontarolo, Roberto

2017-06-15

A systematic and critical review was conducted on bioanalytical methods validated to quantify combinations of antidiabetic agents in human blood. The aim of this article was to verify how the validation process of bioanalytical methods is performed and the quality of the published records. The validation assays were evaluated according to international guidelines. The main problems in the validation process are pointed out and discussed to help researchers to choose methods that are truly reliable and can be successfully applied for their intended use. The combination of oral antidiabetic agents was chosen as these are some of the most studied drugs and several methods are present in the literature. Moreover, this article may be applied to the validation process of all bioanalytical. Copyright © 2017 Elsevier B.V. All rights reserved.

Validation studies and proficiency testing.

PubMed

Ankilam, Elke; Heinze, Petra; Kay, Simon; Van den Eede, Guy; Popping, Bert

2002-01-01

Genetically modified organisms (GMOs) entered the European food market in 1996. Current legislation demands the labeling of food products if they contain <1% GMO, as assessed for each ingredient of the product. To create confidence in the testing methods and to complement enforcement requirements, there is an urgent need for internationally validated methods, which could serve as reference methods. To date, several methods have been submitted to validation trials at an international level; approaches now exist that can be used in different circumstances and for different food matrixes. Moreover, the requirement for the formal validation of methods is clearly accepted; several national and international bodies are active in organizing studies. Further validation studies, especially on the quantitative polymerase chain reaction methods, need to be performed to cover the rising demand for new extraction methods and other background matrixes, as well as for novel GMO constructs.

78 FR 20695 - Walk-Through Metal Detectors and Hand-Held Metal Detectors Test Method Validation

Federal Register 2010, 2011, 2012, 2013, 2014

2013-04-05

... Detectors and Hand-Held Metal Detectors Test Method Validation AGENCY: National Institute of Justice, DOJ... ensure that the test methods in the standards are properly documented, NIJ is requesting proposals (including price quotes) for test method validation efforts from testing laboratories. NIJ is also seeking...

Validity in Mixed Methods Research in Education: The Application of Habermas' Critical Theory

ERIC Educational Resources Information Center

Long, Haiying

2017-01-01

Mixed methods approach has developed into the third methodological movement in educational research. Validity in mixed methods research as an important issue, however, has not been examined as extensively as that of quantitative and qualitative research. Additionally, the previous discussions of validity in mixed methods research focus on research…

Task Validation for the AN/TPQ-36 Radar System

DTIC Science & Technology

1978-09-01

report presents the method and results of a study to validate personnel task descriptions for the new AN/TyP-Jb radar...TP.J-Sb KAPAK SVSTKM CONTENTS i ■ l.t |i- INTRODUCTION t METHOD 2 RESULTS, CONCLUSIONS, AND RECOMMENDATIONS b Task Validation 5 26B MOS... method , results, conclusions, and recommendations of the validation study. The appendixes contain the following: 1. Appendix A contains

How Many Batches Are Needed for Process Validation under the New FDA Guidance?

PubMed

Yang, Harry

2013-01-01

The newly updated FDA Guidance for Industry on Process Validation: General Principles and Practices ushers in a life cycle approach to process validation. While the guidance no longer considers the use of traditional three-batch validation appropriate, it does not prescribe the number of validation batches for a prospective validation protocol, nor does it provide specific methods to determine it. This potentially could leave manufacturers in a quandary. In this paper, I develop a Bayesian method to address the issue. By combining process knowledge gained from Stage 1 Process Design (PD) with expected outcomes of Stage 2 Process Performance Qualification (PPQ), the number of validation batches for PPQ is determined to provide a high level of assurance that the process will consistently produce future batches meeting quality standards. Several examples based on simulated data are presented to illustrate the use of the Bayesian method in helping manufacturers make risk-based decisions for Stage 2 PPQ, and they highlight the advantages of the method over traditional Frequentist approaches. The discussions in the paper lend support for a life cycle and risk-based approach to process validation recommended in the new FDA guidance. The newly updated FDA Guidance for Industry on Process Validation: General Principles and Practices ushers in a life cycle approach to process validation. While the guidance no longer considers the use of traditional three-batch validation appropriate, it does not prescribe the number of validation batches for a prospective validation protocol, nor does it provide specific methods to determine it. This potentially could leave manufacturers in a quandary. In this paper, I develop a Bayesian method to address the issue. By combining process knowledge gained from Stage 1 Process Design (PD) with expected outcomes of Stage 2 Process Performance Qualification (PPQ), the number of validation batches for PPQ is determined to provide a high level of assurance that the process will consistently produce future batches meeting quality standards. Several examples based on simulated data are presented to illustrate the use of the Bayesian method in helping manufacturers make risk-based decisions for Stage 2 PPQ, and THEY highlight the advantages of the method over traditional Frequentist approaches. The discussions in the paper lend support for a life cycle and risk-based approach to process validation recommended in the new FDA guidance.

Random Qualitative Validation: A Mixed-Methods Approach to Survey Validation

ERIC Educational Resources Information Center

Van Duzer, Eric

2012-01-01

The purpose of this paper is to introduce the process and value of Random Qualitative Validation (RQV) in the development and interpretation of survey data. RQV is a method of gathering clarifying qualitative data that improves the validity of the quantitative analysis. This paper is concerned with validity in relation to the participants'…

Content validity across methods of malnutrition assessment in patients with cancer is limited.

PubMed

Sealy, Martine J; Nijholt, Willemke; Stuiver, Martijn M; van der Berg, Marit M; Roodenburg, Jan L N; van der Schans, Cees P; Ottery, Faith D; Jager-Wittenaar, Harriët

2016-08-01

To identify malnutrition assessment methods in cancer patients and assess their content validity based on internationally accepted definitions for malnutrition. Systematic review of studies in cancer patients that operationalized malnutrition as a variable, published since 1998. Eleven key concepts, within the three domains reflected by the malnutrition definitions acknowledged by European Society for Clinical Nutrition and Metabolism (ESPEN) and the American Society for Parenteral and Enteral Nutrition (ASPEN): A: nutrient balance; B: changes in body shape, body area and body composition; and C: function, were used to classify content validity of methods to assess malnutrition. Content validity indices (M-CVIA-C) were calculated per assessment method. Acceptable content validity was defined as M-CVIA-C ≥ 0.80. Thirty-seven assessment methods were identified in the 160 included articles. Mini Nutritional Assessment (M-CVIA-C = 0.72), Scored Patient-Generated Subjective Global Assessment (M-CVIA-C = 0.61), and Subjective Global Assessment (M-CVIA-C = 0.53) scored highest M-CVIA-C. A large number of malnutrition assessment methods are used in cancer research. Content validity of these methods varies widely. None of these assessment methods has acceptable content validity, when compared against a construct based on ESPEN and ASPEN definitions of malnutrition. Copyright © 2016 Elsevier Inc. All rights reserved.

Field validation of the dnph method for aldehydes and ketones. Final report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Workman, G.S.; Steger, J.L.

1996-04-01

A stationary source emission test method for selected aldehydes and ketones has been validated. The method employs a sampling train with impingers containing 2,4-dinitrophenylhydrazine (DNPH) to derivatize the analytes. The resulting hydrazones are recovered and analyzed by high performance liquid chromatography. Nine analytes were studied; the method was validated for formaldehyde, acetaldehyde, propionaldehyde, acetophenone and isophorone. Acrolein, menthyl ethyl ketone, menthyl isobutyl ketone, and quinone did not meet the validation criteria. The study employed the validation techniques described in EPA method 301, which uses train spiking to determine bias, and collocated sampling trains to determine precision. The studies were carriedmore » out at a plywood veneer dryer and a polyester manufacturing plant.« less

Application of 2-Aminothiazoline-4-carboxylic Acid as a Forensic Marker of Cyanide Exposure.

PubMed

Rużycka, Monika; Giebułtowicz, Joanna; Fudalej, Marcin; Krajewski, Paweł; Wroczyński, Piotr

2017-02-20

Cyanides are infamous for their highly poisonous properties. Accidental cyanide poisoning occurs frequently, but occasionally, intentional poisonings also occur. Inhalation of fumes generated by fire may also cause cyanide poisoning. There are many limitations in direct analysis of cyanide. 2-Aminothiazoline-4-carboxylic acid (ATCA), a cyanide metabolite, seems to be the only surrogate that is being used in the detection of cyanide because of its stability and its cyanide-dependent quality in a biological matrix. Unfortunately, toxicokinetic studies on diverse animal models suggest significant interspecies differences; therefore, the attempt to extrapolate animal models to human models may be unsuccessful. The aim of the present study was to evaluate the use of ATCA as a forensic marker of cyanide exposure. For this purpose, post-mortem materials (blood and organs) from fire victims (n = 32) and cyanide-poisoned persons (n = 3) were collected. The distribution of ATCA in organs and its thermal stability were evaluated. The variability of cyanides in a putrid sample and in the context of their long-term and higher temperature stability was established. The presence of ATCA was detected by using an LC-MS/MS method and that of cyanide was detected spectrofluorimetrically. This is the first report on the endogenous ATCA concentrations and the determination of ATCA distribution in tissues of fire victims and cyanide-poisoned persons. It was found that blood and heart had the highest ATCA concentrations. ATCA was observed to be thermally stable even at 90 °C. Even though the cyanide concentration was not elevated in putrid samples, it was unstable during long-term storage and at higher temperature, as expected. The relationship between ATCA and cyanides was also observed. Higher ATCA concentrations were related to increased levels of cyanide in blood and organs (less prominent). ATCA seems to be a reliable forensic marker of exposure to lethal doses of cyanide.

Validating Analytical Methods

ERIC Educational Resources Information Center

Ember, Lois R.

1977-01-01

The procedures utilized by the Association of Official Analytical Chemists (AOAC) to develop, evaluate, and validate analytical methods for the analysis of chemical pollutants are detailed. Methods validated by AOAC are used by the EPA and FDA in their enforcement programs and are granted preferential treatment by the courts. (BT)

A guideline for the validation of likelihood ratio methods used for forensic evidence evaluation.

PubMed

Meuwly, Didier; Ramos, Daniel; Haraksim, Rudolf

2017-07-01

This Guideline proposes a protocol for the validation of forensic evaluation methods at the source level, using the Likelihood Ratio framework as defined within the Bayes' inference model. In the context of the inference of identity of source, the Likelihood Ratio is used to evaluate the strength of the evidence for a trace specimen, e.g. a fingermark, and a reference specimen, e.g. a fingerprint, to originate from common or different sources. Some theoretical aspects of probabilities necessary for this Guideline were discussed prior to its elaboration, which started after a workshop of forensic researchers and practitioners involved in this topic. In the workshop, the following questions were addressed: "which aspects of a forensic evaluation scenario need to be validated?", "what is the role of the LR as part of a decision process?" and "how to deal with uncertainty in the LR calculation?". The questions: "what to validate?" focuses on the validation methods and criteria and "how to validate?" deals with the implementation of the validation protocol. Answers to these questions were deemed necessary with several objectives. First, concepts typical for validation standards [1], such as performance characteristics, performance metrics and validation criteria, will be adapted or applied by analogy to the LR framework. Second, a validation strategy will be defined. Third, validation methods will be described. Finally, a validation protocol and an example of validation report will be proposed, which can be applied to the forensic fields developing and validating LR methods for the evaluation of the strength of evidence at source level under the following propositions. Copyright © 2016. Published by Elsevier B.V.

Validation sampling can reduce bias in health care database studies: an illustration using influenza vaccination effectiveness.

PubMed

Nelson, Jennifer Clark; Marsh, Tracey; Lumley, Thomas; Larson, Eric B; Jackson, Lisa A; Jackson, Michael L

2013-08-01

Estimates of treatment effectiveness in epidemiologic studies using large observational health care databases may be biased owing to inaccurate or incomplete information on important confounders. Study methods that collect and incorporate more comprehensive confounder data on a validation cohort may reduce confounding bias. We applied two such methods, namely imputation and reweighting, to Group Health administrative data (full sample) supplemented by more detailed confounder data from the Adult Changes in Thought study (validation sample). We used influenza vaccination effectiveness (with an unexposed comparator group) as an example and evaluated each method's ability to reduce bias using the control time period before influenza circulation. Both methods reduced, but did not completely eliminate, the bias compared with traditional effectiveness estimates that do not use the validation sample confounders. Although these results support the use of validation sampling methods to improve the accuracy of comparative effectiveness findings from health care database studies, they also illustrate that the success of such methods depends on many factors, including the ability to measure important confounders in a representative and large enough validation sample, the comparability of the full sample and validation sample, and the accuracy with which the data can be imputed or reweighted using the additional validation sample information. Copyright © 2013 Elsevier Inc. All rights reserved.

JaCVAM-organized international validation study of the in vivo rodent alkaline comet assay for the detection of genotoxic carcinogens: I. Summary of pre-validation study results.

PubMed

Uno, Yoshifumi; Kojima, Hajime; Omori, Takashi; Corvi, Raffaella; Honma, Masamistu; Schechtman, Leonard M; Tice, Raymond R; Burlinson, Brian; Escobar, Patricia A; Kraynak, Andrew R; Nakagawa, Yuzuki; Nakajima, Madoka; Pant, Kamala; Asano, Norihide; Lovell, David; Morita, Takeshi; Ohno, Yasuo; Hayashi, Makoto

2015-07-01

The in vivo rodent alkaline comet assay (comet assay) is used internationally to investigate the in vivo genotoxic potential of test chemicals. This assay, however, has not previously been formally validated. The Japanese Center for the Validation of Alternative Methods (JaCVAM), with the cooperation of the U.S. NTP Interagency Center for the Evaluation of Alternative Toxicological Methods (NICEATM)/the Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM), the European Centre for the Validation of Alternative Methods (ECVAM), and the Japanese Environmental Mutagen Society/Mammalian Mutagenesis Study Group (JEMS/MMS), organized an international validation study to evaluate the reliability and relevance of the assay for identifying genotoxic carcinogens, using liver and stomach as target organs. The ultimate goal of this validation effort was to establish an Organisation for Economic Co-operation and Development (OECD) test guideline. The purpose of the pre-validation studies (i.e., Phase 1 through 3), conducted in four or five laboratories with extensive comet assay experience, was to optimize the protocol to be used during the definitive validation study. Copyright © 2015 Elsevier B.V. All rights reserved.

77 FR 35393 - Biennial Progress Report of the Interagency Coordinating Committee on the Validation of...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-06-13

... Committee on the Validation of Alternative Methods (ICCVAM) AGENCY: Division of the National Toxicology... Alternative Toxicological Methods (NICEATM) announces the availability of the Biennial Progress Report 2010-2011: Interagency Coordinating Committee on the Validation of Alternative Methods. The report was...

AOAC Official MethodSM Matrix Extension Validation Study of Assurance GDSTM for the Detection of Salmonella in Selected Spices.

PubMed

Feldsine, Philip; Kaur, Mandeep; Shah, Khyati; Immerman, Amy; Jucker, Markus; Lienau, Andrew

2015-01-01

Assurance GDSTM for Salmonella Tq has been validated according to the AOAC INTERNATIONAL Methods Committee Guidelines for Validation of Microbiological Methods for Food and Environmental Surfaces for the detection of selected foods and environmental surfaces (Official Method of AnalysisSM 2009.03, Performance Tested MethodSM No. 050602). The method also completed AFNOR validation (following the ISO 16140 standard) compared to the reference method EN ISO 6579. For AFNOR, GDS was given a scope covering all human food, animal feed stuff, and environmental surfaces (Certificate No. TRA02/12-01/09). Results showed that Assurance GDS for Salmonella (GDS) has high sensitivity and is equivalent to the reference culture methods for the detection of motile and non-motile Salmonella. As part of the aforementioned validations, inclusivity and exclusivity studies, stability, and ruggedness studies were also conducted. Assurance GDS has 100% inclusivity and exclusivity among the 100 Salmonella serovars and 35 non-Salmonella organisms analyzed. To add to the scope of the Assurance GDS for Salmonella method, a matrix extension study was conducted, following the AOAC guidelines, to validate the application of the method for selected spices, specifically curry powder, cumin powder, and chili powder, for the detection of Salmonella.

Reliability and validity of the AutoCAD software method in lumbar lordosis measurement

PubMed Central

Letafatkar, Amir; Amirsasan, Ramin; Abdolvahabi, Zahra; Hadadnezhad, Malihe

2011-01-01

Objective The aim of this study was to determine the reliability and validity of the AutoCAD software method in lumbar lordosis measurement. Methods Fifty healthy volunteers with a mean age of 23 ± 1.80 years were enrolled. A lumbar lateral radiograph was taken on all participants, and the lordosis was measured according to the Cobb method. Afterward, the lumbar lordosis degree was measured via AutoCAD software and flexible ruler methods. The current study is accomplished in 2 parts: intratester and intertester evaluations of reliability as well as the validity of the flexible ruler and software methods. Results Based on the intraclass correlation coefficient, AutoCAD's reliability and validity in measuring lumbar lordosis were 0.984 and 0.962, respectively. Conclusions AutoCAD showed to be a reliable and valid method to measure lordosis. It is suggested that this method may replace those that are costly and involve health risks, such as radiography, in evaluating lumbar lordosis. PMID:22654681

Critical Values for Lawshe's Content Validity Ratio: Revisiting the Original Methods of Calculation

ERIC Educational Resources Information Center

Ayre, Colin; Scally, Andrew John

2014-01-01

The content validity ratio originally proposed by Lawshe is widely used to quantify content validity and yet methods used to calculate the original critical values were never reported. Methods for original calculation of critical values are suggested along with tables of exact binomial probabilities.

A long-term validation of the modernised DC-ARC-OES solid-sample method.

PubMed

Flórián, K; Hassler, J; Förster, O

2001-12-01

The validation procedure based on ISO 17025 standard has been used to study and illustrate both the longterm stability of the calibration process of the DC-ARC solid sample spectrometric method and the main validation criteria of the method. In the calculation of the validation characteristics depending on the linearity(calibration), also the fulfilment of predetermining criteria such as normality and homoscedasticity was checked. In order to decide whether there are any trends in the time-variation of the analytical signal or not, also the Neumann test of trend was applied and evaluated. Finally, a comparison with similar validation data of the ETV-ICP-OES method was carried out.

VALUE - A Framework to Validate Downscaling Approaches for Climate Change Studies

NASA Astrophysics Data System (ADS)

Maraun, Douglas; Widmann, Martin; Gutiérrez, José M.; Kotlarski, Sven; Chandler, Richard E.; Hertig, Elke; Wibig, Joanna; Huth, Radan; Wilke, Renate A. I.

2015-04-01

VALUE is an open European network to validate and compare downscaling methods for climate change research. VALUE aims to foster collaboration and knowledge exchange between climatologists, impact modellers, statisticians, and stakeholders to establish an interdisciplinary downscaling community. A key deliverable of VALUE is the development of a systematic validation framework to enable the assessment and comparison of both dynamical and statistical downscaling methods. Here, we present the key ingredients of this framework. VALUE's main approach to validation is user-focused: starting from a specific user problem, a validation tree guides the selection of relevant validation indices and performance measures. Several experiments have been designed to isolate specific points in the downscaling procedure where problems may occur: what is the isolated downscaling skill? How do statistical and dynamical methods compare? How do methods perform at different spatial scales? Do methods fail in representing regional climate change? How is the overall representation of regional climate, including errors inherited from global climate models? The framework will be the basis for a comprehensive community-open downscaling intercomparison study, but is intended also to provide general guidance for other validation studies.

VALUE: A framework to validate downscaling approaches for climate change studies

NASA Astrophysics Data System (ADS)

Maraun, Douglas; Widmann, Martin; Gutiérrez, José M.; Kotlarski, Sven; Chandler, Richard E.; Hertig, Elke; Wibig, Joanna; Huth, Radan; Wilcke, Renate A. I.

2015-01-01

VALUE is an open European network to validate and compare downscaling methods for climate change research. VALUE aims to foster collaboration and knowledge exchange between climatologists, impact modellers, statisticians, and stakeholders to establish an interdisciplinary downscaling community. A key deliverable of VALUE is the development of a systematic validation framework to enable the assessment and comparison of both dynamical and statistical downscaling methods. In this paper, we present the key ingredients of this framework. VALUE's main approach to validation is user- focused: starting from a specific user problem, a validation tree guides the selection of relevant validation indices and performance measures. Several experiments have been designed to isolate specific points in the downscaling procedure where problems may occur: what is the isolated downscaling skill? How do statistical and dynamical methods compare? How do methods perform at different spatial scales? Do methods fail in representing regional climate change? How is the overall representation of regional climate, including errors inherited from global climate models? The framework will be the basis for a comprehensive community-open downscaling intercomparison study, but is intended also to provide general guidance for other validation studies.

VDA, a Method of Choosing a Better Algorithm with Fewer Validations

PubMed Central

Kluger, Yuval

2011-01-01

The multitude of bioinformatics algorithms designed for performing a particular computational task presents end-users with the problem of selecting the most appropriate computational tool for analyzing their biological data. The choice of the best available method is often based on expensive experimental validation of the results. We propose an approach to design validation sets for method comparison and performance assessment that are effective in terms of cost and discrimination power. Validation Discriminant Analysis (VDA) is a method for designing a minimal validation dataset to allow reliable comparisons between the performances of different algorithms. Implementation of our VDA approach achieves this reduction by selecting predictions that maximize the minimum Hamming distance between algorithmic predictions in the validation set. We show that VDA can be used to correctly rank algorithms according to their performances. These results are further supported by simulations and by realistic algorithmic comparisons in silico. VDA is a novel, cost-efficient method for minimizing the number of validation experiments necessary for reliable performance estimation and fair comparison between algorithms. Our VDA software is available at http://sourceforge.net/projects/klugerlab/files/VDA/ PMID:22046256

Performance Ratings: Designs for Evaluating Their Validity and Accuracy.

DTIC Science & Technology

1986-07-01

ratees with substantial validity and with little bias due to the ethod for rating. Convergent validity and discriminant validity account for approximately...The expanded research design suggests that purpose for the ratings has little influence on the multitrait-multimethod properties of the ratings...Convergent and discriminant validity again account for substantial differences in the ratings of performance. Little method bias is present; both methods of

Reliability and validity of non-radiographic methods of thoracic kyphosis measurement: a systematic review.

PubMed

Barrett, Eva; McCreesh, Karen; Lewis, Jeremy

2014-02-01

A wide array of instruments are available for non-invasive thoracic kyphosis measurement. Guidelines for selecting outcome measures for use in clinical and research practice recommend that properties such as validity and reliability are considered. This systematic review reports on the reliability and validity of non-invasive methods for measuring thoracic kyphosis. A systematic search of 11 electronic databases located studies assessing reliability and/or validity of non-invasive thoracic kyphosis measurement techniques. Two independent reviewers used a critical appraisal tool to assess the quality of retrieved studies. Data was extracted by the primary reviewer. The results were synthesized qualitatively using a level of evidence approach. 27 studies satisfied the eligibility criteria and were included in the review. The reliability, validity and both reliability and validity were investigated by sixteen, two and nine studies respectively. 17/27 studies were deemed to be of high quality. In total, 15 methods of thoracic kyphosis were evaluated in retrieved studies. All investigated methods showed high (ICC ≥ .7) to very high (ICC ≥ .9) levels of reliability. The validity of the methods ranged from low to very high. The strongest levels of evidence for reliability exists in support of the Debrunner kyphometer, Spinal Mouse and Flexicurve index, and for validity supports the arcometer and Flexicurve index. Further reliability and validity studies are required to strengthen the level of evidence for the remaining methods of measurement. This should be addressed by future research. Copyright © 2013 Elsevier Ltd. All rights reserved.

VALIDATION OF A METHOD FOR ESTIMATING LONG-TERM EXPOSURES BASED ON SHORT-TERM MEASUREMENTS

EPA Science Inventory

A method for estimating long-term exposures from short-term measurements is validated using data from a recent EPA study of exposure to fine particles. The method was developed a decade ago but data to validate it did not exist until recently. In this paper, data from repeated ...

VALIDATION OF A METHOD FOR ESTIMATING LONG-TERM EXPOSURES BASED ON SHORT-TERM MEASUREMENTS

EPA Science Inventory

A method for estimating long-term exposures from short-term measurements is validated using data from a recent EPA study of exposure to fine particles. The method was developed a decade ago but long-term exposure data to validate it did not exist until recently. In this paper, ...

Discriminant Validity Assessment: Use of Fornell & Larcker criterion versus HTMT Criterion

NASA Astrophysics Data System (ADS)

Hamid, M. R. Ab; Sami, W.; Mohmad Sidek, M. H.

2017-09-01

Assessment of discriminant validity is a must in any research that involves latent variables for the prevention of multicollinearity issues. Fornell and Larcker criterion is the most widely used method for this purpose. However, a new method has emerged for establishing the discriminant validity assessment through heterotrait-monotrait (HTMT) ratio of correlations method. Therefore, this article presents the results of discriminant validity assessment using these methods. Data from previous study was used that involved 429 respondents for empirical validation of value-based excellence model in higher education institutions (HEI) in Malaysia. From the analysis, the convergent, divergent and discriminant validity were established and admissible using Fornell and Larcker criterion. However, the discriminant validity is an issue when employing the HTMT criterion. This shows that the latent variables under study faced the issue of multicollinearity and should be looked into for further details. This also implied that the HTMT criterion is a stringent measure that could detect the possible indiscriminant among the latent variables. In conclusion, the instrument which consisted of six latent variables was still lacking in terms of discriminant validity and should be explored further.

Reliability and validity of the AutoCAD software method in lumbar lordosis measurement.

PubMed

Letafatkar, Amir; Amirsasan, Ramin; Abdolvahabi, Zahra; Hadadnezhad, Malihe

2011-12-01

The aim of this study was to determine the reliability and validity of the AutoCAD software method in lumbar lordosis measurement. Fifty healthy volunteers with a mean age of 23 ± 1.80 years were enrolled. A lumbar lateral radiograph was taken on all participants, and the lordosis was measured according to the Cobb method. Afterward, the lumbar lordosis degree was measured via AutoCAD software and flexible ruler methods. The current study is accomplished in 2 parts: intratester and intertester evaluations of reliability as well as the validity of the flexible ruler and software methods. Based on the intraclass correlation coefficient, AutoCAD's reliability and validity in measuring lumbar lordosis were 0.984 and 0.962, respectively. AutoCAD showed to be a reliable and valid method to measure lordosis. It is suggested that this method may replace those that are costly and involve health risks, such as radiography, in evaluating lumbar lordosis.

Cross-Validation of Survival Bump Hunting by Recursive Peeling Methods.

PubMed

Dazard, Jean-Eudes; Choe, Michael; LeBlanc, Michael; Rao, J Sunil

2014-08-01

We introduce a survival/risk bump hunting framework to build a bump hunting model with a possibly censored time-to-event type of response and to validate model estimates. First, we describe the use of adequate survival peeling criteria to build a survival/risk bump hunting model based on recursive peeling methods. Our method called "Patient Recursive Survival Peeling" is a rule-induction method that makes use of specific peeling criteria such as hazard ratio or log-rank statistics. Second, to validate our model estimates and improve survival prediction accuracy, we describe a resampling-based validation technique specifically designed for the joint task of decision rule making by recursive peeling (i.e. decision-box) and survival estimation. This alternative technique, called "combined" cross-validation is done by combining test samples over the cross-validation loops, a design allowing for bump hunting by recursive peeling in a survival setting. We provide empirical results showing the importance of cross-validation and replication.

Cross-Validation of Survival Bump Hunting by Recursive Peeling Methods

PubMed Central

Dazard, Jean-Eudes; Choe, Michael; LeBlanc, Michael; Rao, J. Sunil

2015-01-01

We introduce a survival/risk bump hunting framework to build a bump hunting model with a possibly censored time-to-event type of response and to validate model estimates. First, we describe the use of adequate survival peeling criteria to build a survival/risk bump hunting model based on recursive peeling methods. Our method called “Patient Recursive Survival Peeling” is a rule-induction method that makes use of specific peeling criteria such as hazard ratio or log-rank statistics. Second, to validate our model estimates and improve survival prediction accuracy, we describe a resampling-based validation technique specifically designed for the joint task of decision rule making by recursive peeling (i.e. decision-box) and survival estimation. This alternative technique, called “combined” cross-validation is done by combining test samples over the cross-validation loops, a design allowing for bump hunting by recursive peeling in a survival setting. We provide empirical results showing the importance of cross-validation and replication. PMID:26997922

Validation Methods for Fault-Tolerant avionics and control systems, working group meeting 1

NASA Technical Reports Server (NTRS)

1979-01-01

The proceedings of the first working group meeting on validation methods for fault tolerant computer design are presented. The state of the art in fault tolerant computer validation was examined in order to provide a framework for future discussions concerning research issues for the validation of fault tolerant avionics and flight control systems. The development of positions concerning critical aspects of the validation process are given.

External validation of a Cox prognostic model: principles and methods

PubMed Central

2013-01-01

Background A prognostic model should not enter clinical practice unless it has been demonstrated that it performs a useful role. External validation denotes evaluation of model performance in a sample independent of that used to develop the model. Unlike for logistic regression models, external validation of Cox models is sparsely treated in the literature. Successful validation of a model means achieving satisfactory discrimination and calibration (prediction accuracy) in the validation sample. Validating Cox models is not straightforward because event probabilities are estimated relative to an unspecified baseline function. Methods We describe statistical approaches to external validation of a published Cox model according to the level of published information, specifically (1) the prognostic index only, (2) the prognostic index together with Kaplan-Meier curves for risk groups, and (3) the first two plus the baseline survival curve (the estimated survival function at the mean prognostic index across the sample). The most challenging task, requiring level 3 information, is assessing calibration, for which we suggest a method of approximating the baseline survival function. Results We apply the methods to two comparable datasets in primary breast cancer, treating one as derivation and the other as validation sample. Results are presented for discrimination and calibration. We demonstrate plots of survival probabilities that can assist model evaluation. Conclusions Our validation methods are applicable to a wide range of prognostic studies and provide researchers with a toolkit for external validation of a published Cox model. PMID:23496923

Validation sampling can reduce bias in healthcare database studies: an illustration using influenza vaccination effectiveness

PubMed Central

Nelson, Jennifer C.; Marsh, Tracey; Lumley, Thomas; Larson, Eric B.; Jackson, Lisa A.; Jackson, Michael

2014-01-01

Objective Estimates of treatment effectiveness in epidemiologic studies using large observational health care databases may be biased due to inaccurate or incomplete information on important confounders. Study methods that collect and incorporate more comprehensive confounder data on a validation cohort may reduce confounding bias. Study Design and Setting We applied two such methods, imputation and reweighting, to Group Health administrative data (full sample) supplemented by more detailed confounder data from the Adult Changes in Thought study (validation sample). We used influenza vaccination effectiveness (with an unexposed comparator group) as an example and evaluated each method’s ability to reduce bias using the control time period prior to influenza circulation. Results Both methods reduced, but did not completely eliminate, the bias compared with traditional effectiveness estimates that do not utilize the validation sample confounders. Conclusion Although these results support the use of validation sampling methods to improve the accuracy of comparative effectiveness findings from healthcare database studies, they also illustrate that the success of such methods depends on many factors, including the ability to measure important confounders in a representative and large enough validation sample, the comparability of the full sample and validation sample, and the accuracy with which data can be imputed or reweighted using the additional validation sample information. PMID:23849144

Survey of Verification and Validation Techniques for Small Satellite Software Development

NASA Technical Reports Server (NTRS)

Jacklin, Stephen A.

2015-01-01

The purpose of this paper is to provide an overview of the current trends and practices in small-satellite software verification and validation. This document is not intended to promote a specific software assurance method. Rather, it seeks to present an unbiased survey of software assurance methods used to verify and validate small satellite software and to make mention of the benefits and value of each approach. These methods include simulation and testing, verification and validation with model-based design, formal methods, and fault-tolerant software design with run-time monitoring. Although the literature reveals that simulation and testing has by far the longest legacy, model-based design methods are proving to be useful for software verification and validation. Some work in formal methods, though not widely used for any satellites, may offer new ways to improve small satellite software verification and validation. These methods need to be further advanced to deal with the state explosion problem and to make them more usable by small-satellite software engineers to be regularly applied to software verification. Last, it is explained how run-time monitoring, combined with fault-tolerant software design methods, provides an important means to detect and correct software errors that escape the verification process or those errors that are produced after launch through the effects of ionizing radiation.

Session-RPE Method for Training Load Monitoring: Validity, Ecological Usefulness, and Influencing Factors

PubMed Central

Haddad, Monoem; Stylianides, Georgios; Djaoui, Leo; Dellal, Alexandre; Chamari, Karim

2017-01-01

Purpose: The aim of this review is to (1) retrieve all data validating the Session-rating of perceived exertion (RPE)-method using various criteria, (2) highlight the rationale of this method and its ecological usefulness, and (3) describe factors that can alter RPE and users of this method should take into consideration. Method: Search engines such as SPORTDiscus, PubMed, and Google Scholar databases in the English language between 2001 and 2016 were consulted for the validity and usefulness of the session-RPE method. Studies were considered for further analysis when they used the session-RPE method proposed by Foster et al. in 2001. Participants were athletes of any gender, age, or level of competition. Studies using languages other than English were excluded in the analysis of the validity and reliability of the session-RPE method. Other studies were examined to explain the rationale of the session-RPE method and the origin of RPE. Results: A total of 950 studies cited the Foster et al. study that proposed the session RPE-method. 36 studies have examined the validity and reliability of this proposed method using the modified CR-10. Conclusion: These studies confirmed the validity and good reliability and internal consistency of session-RPE method in several sports and physical activities with men and women of different age categories (children, adolescents, and adults) among various expertise levels. This method could be used as “standing alone” method for training load (TL) monitoring purposes though some recommend to combine it with other physiological parameters as heart rate. PMID:29163016

Patterns of Cognitive Strengths and Weaknesses: Identification Rates, Agreement, and Validity for Learning Disabilities Identification

PubMed Central

Miciak, Jeremy; Fletcher, Jack M.; Stuebing, Karla; Vaughn, Sharon; Tolar, Tammy D.

2014-01-01

Purpose Few empirical investigations have evaluated LD identification methods based on a pattern of cognitive strengths and weaknesses (PSW). This study investigated the reliability and validity of two proposed PSW methods: the concordance/discordance method (C/DM) and cross battery assessment (XBA) method. Methods Cognitive assessment data for 139 adolescents demonstrating inadequate response to intervention was utilized to empirically classify participants as meeting or not meeting PSW LD identification criteria using the two approaches, permitting an analysis of: (1) LD identification rates; (2) agreement between methods; and (3) external validity. Results LD identification rates varied between the two methods depending upon the cut point for low achievement, with low agreement for LD identification decisions. Comparisons of groups that met and did not meet LD identification criteria on external academic variables were largely null, raising questions of external validity. Conclusions This study found low agreement and little evidence of validity for LD identification decisions based on PSW methods. An alternative may be to use multiple measures of academic achievement to guide intervention. PMID:24274155

An individual and dynamic Body Segment Inertial Parameter validation method using ground reaction forces.

PubMed

Hansen, Clint; Venture, Gentiane; Rezzoug, Nasser; Gorce, Philippe; Isableu, Brice

2014-05-07

Over the last decades a variety of research has been conducted with the goal to improve the Body Segment Inertial Parameters (BSIP) estimations but to our knowledge a real validation has never been completely successful, because no ground truth is available. The aim of this paper is to propose a validation method for a BSIP identification method (IM) and to confirm the results by comparing them with recalculated contact forces using inverse dynamics to those obtained by a force plate. Furthermore, the results are compared with the recently proposed estimation method by Dumas et al. (2007). Additionally, the results are cross validated with a high velocity overarm throwing movement. Throughout conditions higher correlations, smaller metrics and smaller RMSE can be found for the proposed BSIP estimation (IM) which shows its advantage compared to recently proposed methods as of Dumas et al. (2007). The purpose of the paper is to validate an already proposed method and to show that this method can be of significant advantage compared to conventional methods. Copyright © 2014 Elsevier Ltd. All rights reserved.

The Value of Qualitative Methods in Social Validity Research

ERIC Educational Resources Information Center

Leko, Melinda M.

2014-01-01

One quality indicator of intervention research is the extent to which the intervention has a high degree of social validity, or practicality. In this study, I drew on Wolf's framework for social validity and used qualitative methods to ascertain five middle schoolteachers' perceptions of the social validity of System 44®--a phonics-based reading…

Unexpected but Most Welcome: Mixed Methods for the Validation and Revision of the Participatory Evaluation Measurement Instrument

ERIC Educational Resources Information Center

Daigneault, Pierre-Marc; Jacob, Steve

2014-01-01

Although combining methods is nothing new, more contributions about why and how to mix methods for validation purposes are needed. This article presents a case of validating the inferences drawn from the Participatory Evaluation Measurement Instrument, an instrument that purports to measure stakeholder participation in evaluation. Although the…

Use of the Method of Triads in the Validation of Sodium and Potassium Intake in the Brazilian Longitudinal Study of Adult Health (ELSA-Brasil)

PubMed Central

Pereira, Taísa Sabrina Silva; Cade, Nágela Valadão; Mill, José Geraldo; Sichieri, Rosely; Molina, Maria del Carmen Bisi

2016-01-01

Introduction Biomarkers are a good choice to be used in the validation of food frequency questionnaire due to the independence of their random errors. Objective To assess the validity of the potassium and sodium intake estimated using the Food Frequency Questionnaire ELSA-Brasil. Subjects/Methods A subsample of participants in the ELSA-Brasil cohort was included in this study in 2009. Sodium and potassium intake were estimated using three methods: Semi-quantitative food frequency questionnaire, 12-hour nocturnal urinary excretion and three 24-hour food records. Correlation coefficients were calculated between the methods, and the validity coefficient was calculated using the method of triads. The 95% confidence intervals for the validity coefficient were estimated using bootstrap sampling. Exact and adjacent agreement and disagreement of the estimated sodium and potassium intake quintiles were compared among three methods. Results The sample consisted of 246 participants, aged 53±8 years, 52% of women. Validity coefficient for sodium were considered weak (рfood frequency questionnaire actual intake = 0.37 and рbiomarker actual intake = 0.21) and moderate (рfood records actual intake 0.56). The validity coefficient were higher for potassium (рfood frequency questionnaire actual intake = 0.60; рbiomarker actual intake = 0.42; рfood records actual intake = 0.79). Conclusions: The Food Frequency Questionnaire ELSA-Brasil showed good validity in estimating potassium intake in epidemiological studies. For sodium validity was weak, likely due to the non-quantification of the added salt to prepared food. PMID:28030625

Increased efficacy for in-house validation of real-time PCR GMO detection methods.

PubMed

Scholtens, I M J; Kok, E J; Hougs, L; Molenaar, B; Thissen, J T N M; van der Voet, H

2010-03-01

To improve the efficacy of the in-house validation of GMO detection methods (DNA isolation and real-time PCR, polymerase chain reaction), a study was performed to gain insight in the contribution of the different steps of the GMO detection method to the repeatability and in-house reproducibility. In the present study, 19 methods for (GM) soy, maize canola and potato were validated in-house of which 14 on the basis of an 8-day validation scheme using eight different samples and five on the basis of a more concise validation protocol. In this way, data was obtained with respect to the detection limit, accuracy and precision. Also, decision limits were calculated for declaring non-conformance (>0.9%) with 95% reliability. In order to estimate the contribution of the different steps in the GMO analysis to the total variation variance components were estimated using REML (residual maximum likelihood method). From these components, relative standard deviations for repeatability and reproducibility (RSD(r) and RSD(R)) were calculated. The results showed that not only the PCR reaction but also the factors 'DNA isolation' and 'PCR day' are important factors for the total variance and should therefore be included in the in-house validation. It is proposed to use a statistical model to estimate these factors from a large dataset of initial validations so that for similar GMO methods in the future, only the PCR step needs to be validated. The resulting data are discussed in the light of agreed European criteria for qualified GMO detection methods.

Validation of Milliflex® Quantum for Bioburden Testing of Pharmaceutical Products.

PubMed

Gordon, Oliver; Goverde, Marcel; Staerk, Alexandra; Roesti, David

2017-01-01

This article reports the validation strategy used to demonstrate that the Milliflex ® Quantum yielded non-inferior results to the traditional bioburden method. It was validated according to USP <1223>, European Pharmacopoeia 5.1.6, and Parenteral Drug Association Technical Report No. 33 and comprised the validation parameters robustness, ruggedness, repeatability, specificity, limit of detection and quantification, accuracy, precision, linearity, range, and equivalence in routine operation. For the validation, a combination of pharmacopeial ATCC strains as well as a broad selection of in-house isolates were used. In-house isolates were used in stressed state. Results were statistically evaluated regarding the pharmacopeial acceptance criterion of ≥70% recovery compared to the traditional method. Post-hoc test power calculations verified the appropriateness of the used sample size to detect such a difference. Furthermore, equivalence tests verified non-inferiority of the rapid method as compared to the traditional method. In conclusion, the rapid bioburden on basis of the Milliflex ® Quantum was successfully validated as alternative method to the traditional bioburden test. LAY ABSTRACT: Pharmaceutical drug products must fulfill specified quality criteria regarding their microbial content in order to ensure patient safety. Drugs that are delivered into the body via injection, infusion, or implantation must be sterile (i.e., devoid of living microorganisms). Bioburden testing measures the levels of microbes present in the bulk solution of a drug before sterilization, and thus it provides important information for manufacturing a safe product. In general, bioburden testing has to be performed using the methods described in the pharmacopoeias (membrane filtration or plate count). These methods are well established and validated regarding their effectiveness; however, the incubation time required to visually identify microbial colonies is long. Thus, alternative methods that detect microbial contamination faster will improve control over the manufacturing process and speed up product release. Before alternative methods may be used, they must undergo a side-by-side comparison with pharmacopeial methods. In this comparison, referred to as validation, it must be shown in a statistically verified manner that the effectiveness of the alternative method is at least equivalent to that of the pharmacopeial methods. Here we describe the successful validation of an alternative bioburden testing method based on fluorescent staining of growing microorganisms applying the Milliflex ® Quantum system by MilliporeSigma. © PDA, Inc. 2017.

Validation and Clinical Evaluation of a Novel Method To Measure Miltefosine in Leishmaniasis Patients Using Dried Blood Spot Sample Collection

PubMed Central

Rosing, H.; Hillebrand, M. J. X.; Blesson, S.; Mengesha, B.; Diro, E.; Hailu, A.; Schellens, J. H. M.; Beijnen, J. H.

2016-01-01

To facilitate future pharmacokinetic studies of combination treatments against leishmaniasis in remote regions in which the disease is endemic, a simple cheap sampling method is required for miltefosine quantification. The aims of this study were to validate a liquid chromatography-tandem mass spectrometry method to quantify miltefosine in dried blood spot (DBS) samples and to validate its use with Ethiopian patients with visceral leishmaniasis (VL). Since hematocrit (Ht) levels are typically severely decreased in VL patients, returning to normal during treatment, the method was evaluated over a range of clinically relevant Ht values. Miltefosine was extracted from DBS samples using a simple method of pretreatment with methanol, resulting in >97% recovery. The method was validated over a calibration range of 10 to 2,000 ng/ml, and accuracy and precision were within ±11.2% and ≤7.0% (≤19.1% at the lower limit of quantification), respectively. The method was accurate and precise for blood spot volumes between 10 and 30 μl and for Ht levels of 20 to 35%, although a linear effect of Ht levels on miltefosine quantification was observed in the bioanalytical validation. DBS samples were stable for at least 162 days at 37°C. Clinical validation of the method using paired DBS and plasma samples from 16 VL patients showed a median observed DBS/plasma miltefosine concentration ratio of 0.99, with good correlation (Pearson's r = 0.946). Correcting for patient-specific Ht levels did not further improve the concordance between the sampling methods. This successfully validated method to quantify miltefosine in DBS samples was demonstrated to be a valid and practical alternative to venous blood sampling that can be applied in future miltefosine pharmacokinetic studies with leishmaniasis patients, without Ht correction. PMID:26787691

Beware of external validation! - A Comparative Study of Several Validation Techniques used in QSAR Modelling.

PubMed

Majumdar, Subhabrata; Basak, Subhash C

2018-04-26

Proper validation is an important aspect of QSAR modelling. External validation is one of the widely used validation methods in QSAR where the model is built on a subset of the data and validated on the rest of the samples. However, its effectiveness for datasets with a small number of samples but large number of predictors remains suspect. Calculating hundreds or thousands of molecular descriptors using currently available software has become the norm in QSAR research, owing to computational advances in the past few decades. Thus, for n chemical compounds and p descriptors calculated for each molecule, the typical chemometric dataset today has high value of p but small n (i.e. n < p). Motivated by the evidence of inadequacies of external validation in estimating the true predictive capability of a statistical model in recent literature, this paper performs an extensive and comparative study of this method with several other validation techniques. We compared four validation methods: leave-one-out, K-fold, external and multi-split validation, using statistical models built using the LASSO regression, which simultaneously performs variable selection and modelling. We used 300 simulated datasets and one real dataset of 95 congeneric amine mutagens for this evaluation. External validation metrics have high variation among different random splits of the data, hence are not recommended for predictive QSAR models. LOO has the overall best performance among all validation methods applied in our scenario. Results from external validation are too unstable for the datasets we analyzed. Based on our findings, we recommend using the LOO procedure for validating QSAR predictive models built on high-dimensional small-sample data. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

A Generalized Pivotal Quantity Approach to Analytical Method Validation Based on Total Error.

PubMed

Yang, Harry; Zhang, Jianchun

2015-01-01

The primary purpose of method validation is to demonstrate that the method is fit for its intended use. Traditionally, an analytical method is deemed valid if its performance characteristics such as accuracy and precision are shown to meet prespecified acceptance criteria. However, these acceptance criteria are not directly related to the method's intended purpose, which is usually a gurantee that a high percentage of the test results of future samples will be close to their true values. Alternate "fit for purpose" acceptance criteria based on the concept of total error have been increasingly used. Such criteria allow for assessing method validity, taking into account the relationship between accuracy and precision. Although several statistical test methods have been proposed in literature to test the "fit for purpose" hypothesis, the majority of the methods are not designed to protect the risk of accepting unsuitable methods, thus having the potential to cause uncontrolled consumer's risk. In this paper, we propose a test method based on generalized pivotal quantity inference. Through simulation studies, the performance of the method is compared to five existing approaches. The results show that both the new method and the method based on β-content tolerance interval with a confidence level of 90%, hereafter referred to as the β-content (0.9) method, control Type I error and thus consumer's risk, while the other existing methods do not. It is further demonstrated that the generalized pivotal quantity method is less conservative than the β-content (0.9) method when the analytical methods are biased, whereas it is more conservative when the analytical methods are unbiased. Therefore, selection of either the generalized pivotal quantity or β-content (0.9) method for an analytical method validation depends on the accuracy of the analytical method. It is also shown that the generalized pivotal quantity method has better asymptotic properties than all of the current methods. Analytical methods are often used to ensure safety, efficacy, and quality of medicinal products. According to government regulations and regulatory guidelines, these methods need to be validated through well-designed studies to minimize the risk of accepting unsuitable methods. This article describes a novel statistical test for analytical method validation, which provides better protection for the risk of accepting unsuitable analytical methods. © PDA, Inc. 2015.

VALIDATION OF ANALYTICAL METHODS AND INSTRUMENTATION FOR BERYLLIUM MEASUREMENT: REVIEW AND SUMMARY OF AVAILABLE GUIDES, PROCEDURES, AND PROTOCOLS

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ekechukwu, A

Method validation is the process of evaluating whether an analytical method is acceptable for its intended purpose. For pharmaceutical methods, guidelines from the United States Pharmacopeia (USP), International Conference on Harmonisation (ICH), and the United States Food and Drug Administration (USFDA) provide a framework for performing such valications. In general, methods for regulatory compliance must include studies on specificity, linearity, accuracy, precision, range, detection limit, quantitation limit, and robustness. Elements of these guidelines are readily adapted to the issue of validation for beryllium sampling and analysis. This document provides a listing of available sources which can be used to validatemore » analytical methods and/or instrumentation for beryllium determination. A literature review was conducted of available standard methods and publications used for method validation and/or quality control. A comprehensive listing of the articles, papers and books reviewed is given in the Appendix. Available validation documents and guides are listed therein; each has a brief description of application and use. In the referenced sources, there are varying approches to validation and varying descriptions of the valication process at different stages in method development. This discussion focuses on valication and verification of fully developed methods and instrumentation that have been offered up for use or approval by other laboratories or official consensus bodies such as ASTM International, the International Standards Organization (ISO) and the Association of Official Analytical Chemists (AOAC). This review was conducted as part of a collaborative effort to investigate and improve the state of validation for measuring beryllium in the workplace and the environment. Documents and publications from the United States and Europe are included. Unless otherwise specified, all referenced documents were published in English.« less

Do placebo based validation standards mimic real batch products behaviour? Case studies.

PubMed

Bouabidi, A; Talbi, M; Bouklouze, A; El Karbane, M; Bourichi, H; El Guezzar, M; Ziemons, E; Hubert, Ph; Rozet, E

2011-06-01

Analytical methods validation is a mandatory step to evaluate the ability of developed methods to provide accurate results for their routine application. Validation usually involves validation standards or quality control samples that are prepared in placebo or reconstituted matrix made of a mixture of all the ingredients composing the drug product except the active substance or the analyte under investigation. However, one of the main concerns that can be made with this approach is that it may lack an important source of variability that come from the manufacturing process. The question that remains at the end of the validation step is about the transferability of the quantitative performance from validation standards to real authentic drug product samples. In this work, this topic is investigated through three case studies. Three analytical methods were validated using the commonly spiked placebo validation standards at several concentration levels as well as using samples coming from authentic batch samples (tablets and syrups). The results showed that, depending on the type of response function used as calibration curve, there were various degrees of differences in the results accuracy obtained with the two types of samples. Nonetheless the use of spiked placebo validation standards was showed to mimic relatively well the quantitative behaviour of the analytical methods with authentic batch samples. Adding these authentic batch samples into the validation design may help the analyst to select and confirm the most fit for purpose calibration curve and thus increase the accuracy and reliability of the results generated by the method in routine application. Copyright © 2011 Elsevier B.V. All rights reserved.

Method development and validation of potent pyrimidine derivative by UV-VIS spectrophotometer.

PubMed

Chaudhary, Anshu; Singh, Anoop; Verma, Prabhakar Kumar

2014-12-01

A rapid and sensitive ultraviolet-visible (UV-VIS) spectroscopic method was developed for the estimation of pyrimidine derivative 6-Bromo-3-(6-(2,6-dichlorophenyl)-2-(morpolinomethylamino) pyrimidine4-yl) -2H-chromen-2-one (BT10M) in bulk form. Pyrimidine derivative was monitored at 275 nm with UV detection, and there is no interference of diluents at 275 nm. The method was found to be linear in the range of 50 to 150 μg/ml. The accuracy and precision were determined and validated statistically. The method was validated as a guideline. The results showed that the proposed method is suitable for the accurate, precise, and rapid determination of pyrimidine derivative. Graphical Abstract Method development and validation of potent pyrimidine derivative by UV spectroscopy.

Curcumin-functionalized silk materials for enhancing adipogenic differentiation of bone marrow-derived human mesenchymal stem cells

PubMed Central

Li, Chunmei; Luo, Tingting; Zheng, Zhaozhu; Murphy, Amanda R.; Wang, Xiaoqin; Kaplan, David L.

2014-01-01

Curcumin, a natural phenolic compound derived from the plant Curcuma longa, was physically entrapped and stabilized in silk hydrogel films and its influence on human bone marrow-derived mesenchymal stem cells (hBMSCs) was assessed related to adipogenic differentiation. The presence of curcumin significantly reduced silk gelation time and changed the porous morphology of gel matrix, but did not change the formation of silk beta-sheet structure. Based on spectrofluorimetric analysis, curcumin likely interacted with hydrophobic residues in silk, interacting with the beta-sheet domains formed in the hydrogels. The antioxidant activity of silk film-associated curcumin remained functional over at least one month in both the dry and hydrated state. Negligible curcumin was released from silk hydrogel films over 48 hours incubation in aqueous solution. For hBMSCs cultured on silk films containing more than 0.25 mg/mL curcumin, cell proliferation was inhibited while adipogenesis was significantly promoted based on transcripts as well as oil red O staining. When hBMSCs were cultured in media containing free curcumin, both proliferation and adipogenesis of hBMSCs were inhibited when curcumin concentrations exceeded 5 μM, which is more than 1,000-times higher than the level of curcumin released from the films in aqueous solution. Thus, silk film-associated curcumin exhibited different effects on hBMSC proliferation and differentiation when compared to curcumin in solution. PMID:25132274

Synthesis and antiplasmodial activity of betulinic acid and ursolic acid analogues.

PubMed

Innocente, Adrine M; Silva, Gloria N S; Cruz, Laura Nogueira; Moraes, Miriam S; Nakabashi, Myna; Sonnet, Pascal; Gosmann, Grace; Garcia, Célia R S; Gnoatto, Simone C B

2012-10-12

More than 40% of the World population is at risk of contracting malaria, which affects primarily poor populations in tropical and subtropical areas. Antimalarial pharmacotherapy has utilised plant-derived products such as quinine and artemisinin as well as their derivatives. However, worldwide use of these antimalarials has caused the spread of resistant parasites, resulting in increased malaria morbidity and mortality. Considering that the literature has demonstrated the antimalarial potential of triterpenes, specially betulinic acid (1) and ursolic acid (2), this study investigated the antimalarial activity against P. falciparum chloroquine-sensitive 3D7 strain of some new derivatives of 1 and 2 with modifications at C-3 and C-28. The antiplasmodial study employed flow cytometry and spectrofluorimetric analyses using YOYO-1, dihydroethidium and Fluo4/AM for staining. Among the six analogues obtained, compounds 1c and 2c showed excellent activity (IC₅₀ = 220 and 175 nM, respectively) while 1a and b demonstrated good activity (IC₅₀ = 4 and 5 μM, respectively). After cytotoxicity evaluation against HEK293T cells, 1a was not toxic, while 1c and 2c showed IC₅₀ of 4 μM and a selectivity index (SI) value of 18 and 23, respectively. Moreover, compound 2c, which presents the best antiplasmodial activity, is involved in the calcium-regulated pathway(s).

Novel alanines bearing a heteroaromatic side chain: synthesis and studies on fluorescent chemosensing of metal cations with biological relevance.

PubMed

Ferreira, Rosa Cristina M; Raposo, Maria Manuela M; Costa, Susana P G

2018-06-01

A family of novel thienylbenzoxazol-5-yl-L-alanines, consisting of an alanine core bearing a benzoxazole at the side chain with a thiophene ring at position 2, substituted with different (hetero)aryl substituents, was synthesised to study the tuning of the photophysical and chemosensory properties of the resulting compounds. These novel heterocyclic alanines 3a-f and a series of structurally related bis-thienylbenzoxazolyl-alanines 3g-j were evaluated for the first time in the recognition of selected metal cations with environmental, medicinal and analytical interest such as Co 2+ , Cu 2+ , Zn 2+ and Ni 2+ , in acetonitrile solution, with the heterocycles at the side chain acting simultaneously as the coordinating and reporting units, via fluorescence changes. This behaviour can be explained by the involvement of the electron donor heteroatoms in the recognition event, through complexation of the metal cations. The spectrofluorimetric titrations showed that thienylbenzoxazolyl-alanines 3a-j and 4a,b were non-selective fluorimetric chemosensors for the above-mentioned cations, with the best results being obtained for the interaction of Cu 2+ with bis-alanine 3j and deprotected alanines 4a,b. The encouraging photophysical and metal ion sensing properties of these thienylbenzoxazolyl-alanines suggest that they can be used to obtain bioinspired fluorescent reporters for metal ion such as peptides/proteins with chemosensory/probing ability.

Methylene blue improves mitochondrial respiration and decreases oxidative stress in a substrate-dependent manner in diabetic rat hearts.

PubMed

Duicu, Oana M; Privistirescu, Andreea; Wolf, Adrian; Petruş, Alexandra; Dănilă, Maria D; Raţiu, Corina D; Muntean, Danina M; Sturza, Adrian

2017-11-01

Diabetic cardiomyopathy has been systematically associated with compromised mitochondrial energetics and increased generation of reactive oxygen species (ROS) that underlie its progression to heart failure. Methylene blue is a redox drug with reported protective effects mainly on brain mitochondria. The purpose of the present study was to characterize the effects of acute administration of methylene blue on mitochondrial respiration, H 2 O 2 production, and calcium sensitivity in rat heart mitochondria isolated from healthy and 2 months (streptozotocin-induced) diabetic rats. Mitochondrial respiratory function was assessed by high-resolution respirometry. H 2 O 2 production and calcium retention capacity were measured spectrofluorimetrically. The addition of methylene blue (0.1 μmol·L -1 ) elicited an increase in oxygen consumption of mitochondria energized with complex I and II substrates in both normal and diseased mitochondria. Interestingly, methylene blue elicited a significant increase in H 2 O 2 release in the presence of complex I substrates (glutamate and malate), but had an opposite effect in mitochondria energized with complex II substrate (succinate). No changes in the calcium retention capacity of healthy or diabetic mitochondria were found in the presence of methylene blue. In conclusion, in cardiac mitochondria isolated from diabetic and nondiabetic rat hearts, methylene blue improved respiratory function and elicited a dichotomic, substrate-dependent effect on ROS production.

Initial Validation of Ballistic Shock Transducers

DTIC Science & Technology

2017-06-05

objective of this TOP is to describe various methods and instrumentation used in the initial validation of accelerometers to be used in both Ballistic...MILITARY STANDARD 810G CHANGE NOTICE 1, METHOD 522.2, SECTION 4.4.2 ....................................... A-1 B. GLOSSARY...Procedure (TOP) is to describe various methods and instrumentation used in the initial validation of accelerometers to be used in both Ballistic

Experimental validation of structural optimization methods

NASA Technical Reports Server (NTRS)

Adelman, Howard M.

1992-01-01

The topic of validating structural optimization methods by use of experimental results is addressed. The need for validating the methods as a way of effecting a greater and an accelerated acceptance of formal optimization methods by practicing engineering designers is described. The range of validation strategies is defined which includes comparison of optimization results with more traditional design approaches, establishing the accuracy of analyses used, and finally experimental validation of the optimization results. Examples of the use of experimental results to validate optimization techniques are described. The examples include experimental validation of the following: optimum design of a trussed beam; combined control-structure design of a cable-supported beam simulating an actively controlled space structure; minimum weight design of a beam with frequency constraints; minimization of the vibration response of helicopter rotor blade; minimum weight design of a turbine blade disk; aeroelastic optimization of an aircraft vertical fin; airfoil shape optimization for drag minimization; optimization of the shape of a hole in a plate for stress minimization; optimization to minimize beam dynamic response; and structural optimization of a low vibration helicopter rotor.

Use of Bayesian Networks to Probabilistically Model and Improve the Likelihood of Validation of Microarray Findings by RT-PCR

PubMed Central

English, Sangeeta B.; Shih, Shou-Ching; Ramoni, Marco F.; Smith, Lois E.; Butte, Atul J.

2014-01-01

Though genome-wide technologies, such as microarrays, are widely used, data from these methods are considered noisy; there is still varied success in downstream biological validation. We report a method that increases the likelihood of successfully validating microarray findings using real time RT-PCR, including genes at low expression levels and with small differences. We use a Bayesian network to identify the most relevant sources of noise based on the successes and failures in validation for an initial set of selected genes, and then improve our subsequent selection of genes for validation based on eliminating these sources of noise. The network displays the significant sources of noise in an experiment, and scores the likelihood of validation for every gene. We show how the method can significantly increase validation success rates. In conclusion, in this study, we have successfully added a new automated step to determine the contributory sources of noise that determine successful or unsuccessful downstream biological validation. PMID:18790084

A validated ultra high pressure liquid chromatographic method for qualification and quantification of folic acid in pharmaceutical preparations.

PubMed

Deconinck, E; Crevits, S; Baten, P; Courselle, P; De Beer, J

2011-04-05

A fully validated UHPLC method for the identification and quantification of folic acid in pharmaceutical preparations was developed. The starting conditions for the development were calculated starting from the HPLC conditions of a validated method. These start conditions were tested on four different UHPLC columns: Grace Vision HT™ C18-P, C18, C18-HL and C18-B (2 mm × 100 mm, 1.5 μm). After selection of the stationary phase, the method was further optimised by testing two aqueous and two organic phases and by adapting to a gradient method. The obtained method was fully validated based on its measurement uncertainty (accuracy profile) and robustness tests. A UHPLC method was obtained for the identification and quantification of folic acid in pharmaceutical preparations, which will cut analysis times and solvent consumption. Copyright © 2010 Elsevier B.V. All rights reserved.

Probability of Detection (POD) as a statistical model for the validation of qualitative methods.

PubMed

Wehling, Paul; LaBudde, Robert A; Brunelle, Sharon L; Nelson, Maria T

2011-01-01

A statistical model is presented for use in validation of qualitative methods. This model, termed Probability of Detection (POD), harmonizes the statistical concepts and parameters between quantitative and qualitative method validation. POD characterizes method response with respect to concentration as a continuous variable. The POD model provides a tool for graphical representation of response curves for qualitative methods. In addition, the model allows comparisons between candidate and reference methods, and provides calculations of repeatability, reproducibility, and laboratory effects from collaborative study data. Single laboratory study and collaborative study examples are given.

Specification and Preliminary Validation of IAT (Integrated Analysis Techniques) Methods: Executive Summary.

DTIC Science & Technology

1985-03-01

conceptual framwork , and preliminary validation of IAT concepts. Planned work for FY85, including more extensive validation, is also described. 20...Developments: Required Capabilities .... ......... 10 2-1 IAT Conceptual Framework - FY85 (FEO) ..... ........... 11 2-2 Recursive Nature of Decomposition...approach: 1) Identify needs & requirements for IAT. 2) Develop IAT conceptual framework. 3) Validate IAT methods. 4) Develop applications materials. To

Methods to compute reliabilities for genomic predictions of feed intake

USDA-ARS?s Scientific Manuscript database

For new traits without historical reference data, cross-validation is often the preferred method to validate reliability (REL). Time truncation is less useful because few animals gain substantial REL after the truncation point. Accurate cross-validation requires separating genomic gain from pedigree...

Use of multiple cluster analysis methods to explore the validity of a community outcomes concept map.

PubMed

Orsi, Rebecca

2017-02-01

Concept mapping is now a commonly-used technique for articulating and evaluating programmatic outcomes. However, research regarding validity of knowledge and outcomes produced with concept mapping is sparse. The current study describes quantitative validity analyses using a concept mapping dataset. We sought to increase the validity of concept mapping evaluation results by running multiple cluster analysis methods and then using several metrics to choose from among solutions. We present four different clustering methods based on analyses using the R statistical software package: partitioning around medoids (PAM), fuzzy analysis (FANNY), agglomerative nesting (AGNES) and divisive analysis (DIANA). We then used the Dunn and Davies-Bouldin indices to assist in choosing a valid cluster solution for a concept mapping outcomes evaluation. We conclude that the validity of the outcomes map is high, based on the analyses described. Finally, we discuss areas for further concept mapping methods research. Copyright © 2016 Elsevier Ltd. All rights reserved.

The NIH analytical methods and reference materials program for dietary supplements.

PubMed

Betz, Joseph M; Fisher, Kenneth D; Saldanha, Leila G; Coates, Paul M

2007-09-01

Quality of botanical products is a great uncertainty that consumers, clinicians, regulators, and researchers face. Definitions of quality abound, and include specifications for sanitation, adventitious agents (pesticides, metals, weeds), and content of natural chemicals. Because dietary supplements (DS) are often complex mixtures, they pose analytical challenges and method validation may be difficult. In response to product quality concerns and the need for validated and publicly available methods for DS analysis, the US Congress directed the Office of Dietary Supplements (ODS) at the National Institutes of Health (NIH) to accelerate an ongoing methods validation process, and the Dietary Supplements Methods and Reference Materials Program was created. The program was constructed from stakeholder input and incorporates several federal procurement and granting mechanisms in a coordinated and interlocking framework. The framework facilitates validation of analytical methods, analytical standards, and reference materials.

Nonclinical dose formulation analysis method validation and sample analysis.

PubMed

Whitmire, Monica Lee; Bryan, Peter; Henry, Teresa R; Holbrook, John; Lehmann, Paul; Mollitor, Thomas; Ohorodnik, Susan; Reed, David; Wietgrefe, Holly D

2010-12-01

Nonclinical dose formulation analysis methods are used to confirm test article concentration and homogeneity in formulations and determine formulation stability in support of regulated nonclinical studies. There is currently no regulatory guidance for nonclinical dose formulation analysis method validation or sample analysis. Regulatory guidance for the validation of analytical procedures has been developed for drug product/formulation testing; however, verification of the formulation concentrations falls under the framework of GLP regulations (not GMP). The only current related regulatory guidance is the bioanalytical guidance for method validation. The fundamental parameters for bioanalysis and formulation analysis validations that overlap include: recovery, accuracy, precision, specificity, selectivity, carryover, sensitivity, and stability. Divergence in bioanalytical and drug product validations typically center around the acceptance criteria used. As the dose formulation samples are not true "unknowns", the concept of quality control samples that cover the entire range of the standard curve serving as the indication for the confidence in the data generated from the "unknown" study samples may not always be necessary. Also, the standard bioanalytical acceptance criteria may not be directly applicable, especially when the determined concentration does not match the target concentration. This paper attempts to reconcile the different practices being performed in the community and to provide recommendations of best practices and proposed acceptance criteria for nonclinical dose formulation method validation and sample analysis.

Prediction of adult height in girls: the Beunen-Malina-Freitas method.

PubMed

Beunen, Gaston P; Malina, Robert M; Freitas, Duarte L; Thomis, Martine A; Maia, José A; Claessens, Albrecht L; Gouveia, Elvio R; Maes, Hermine H; Lefevre, Johan

2011-12-01

The purpose of this study was to validate and cross-validate the Beunen-Malina-Freitas method for non-invasive prediction of adult height in girls. A sample of 420 girls aged 10-15 years from the Madeira Growth Study were measured at yearly intervals and then 8 years later. Anthropometric dimensions (lengths, breadths, circumferences, and skinfolds) were measured; skeletal age was assessed using the Tanner-Whitehouse 3 method and menarcheal status (present or absent) was recorded. Adult height was measured and predicted using stepwise, forward, and maximum R (2) regression techniques. Multiple correlations, mean differences, standard errors of prediction, and error boundaries were calculated. A sample of the Leuven Longitudinal Twin Study was used to cross-validate the regressions. Age-specific coefficients of determination (R (2)) between predicted and measured adult height varied between 0.57 and 0.96, while standard errors of prediction varied between 1.1 and 3.9 cm. The cross-validation confirmed the validity of the Beunen-Malina-Freitas method in girls aged 12-15 years, but at lower ages the cross-validation was less consistent. We conclude that the Beunen-Malina-Freitas method is valid for the prediction of adult height in girls aged 12-15 years. It is applicable to European populations or populations of European ancestry.

Multifactorial biogeochemical monitoring of linden alley in Moscow

NASA Astrophysics Data System (ADS)

Ermakov, Vadim; Khushvakhtova, Sabsbakhor; Tyutikov, Sergey; Danilova, Valentina; Roca, Núria; Bech, Jaume

2015-04-01

The ecological and biogeochemical assessment of the linden alley within the Kosygin Street was conducted by means of an integrated comparative study of soils, their chemical composition and morphological parameters of leaf linden. For this purpose 5 points were tested within the linden alley and 5 other points outside the highway. In soils, water extract of soil, leaf linden the content of Cu, Pb, Mn, Fe, Cd, Zn, As, Ni, Co Mo, Cr and Se were determined by AAS and spectrofluorimetric method [1]. Macrocomponents (Ca, Mg, K, Na, P, sulphates, chlorides), pH and total mineralization of water soil extract were measured by generally accepted methods. Thio-containing compounds in the leaves were determined by HPLC-NAM spectrofluorometry [2]. On level content of trace elements the soils of "contaminated" points different from background more high concentrations of lead, manganese, iron, selenium, strontium and low level of zinc. Leaf of linden from contaminated sites characterized by an increase of lead, copper, iron, zinc, arsenic, chromium, and a sharp decrease in the level of manganese and strontium. Analysis of the aqueous extracts of the soil showed a slight decrease in the pH value in the "control" points and lower content of calcium, magnesium, potassium, sodium and total mineralization of the water soil extract. The phytochelatins test in the leaves of linden was weakly effective and the degree of asymmetry of leaf lamina too. The most differences between the variants were marked by the degree of pathology leaves (chlorosis and necrosis) and the content of pigments (chlorophyll and carotene). The data obtained reflect the impact of the application of de-icing salts and automobile emissions. References 1. Ermakov V.V., Danilova V.N., Khyshvakhtova S.D. Application of HPLC-NAM spectrofluorimtry to determination of sulfur-containing compounds in the environmental objects// Science of the biosphere: Innovation. Moscow State University by M.V. Lomonosov, 2014. P. 10-12. 2. Ermakov V.V., Tyutikov S.F., Khushvakhtova S.D., Danilova V.N., Boev V.N., Barabanschikova R.N., Chudinova E.A. Peculiarities of quantitative determination of selenium in biological materials// Bulletin of the Tyumen State University Press, 2010, 3, 206-214. Supported by the Russian Foundation for Basic Research, grant number 15-05-00279a

Evaluation of coarse scale land surface remote sensing albedo product over rugged terrain

NASA Astrophysics Data System (ADS)

Wen, J.; Xinwen, L.; You, D.; Dou, B.

2017-12-01

Satellite derived Land surface albedo is an essential climate variable which controls the earth energy budget and it can be used in applications such as climate change, hydrology, and numerical weather prediction. The accuracy and uncertainty of surface albedo products should be evaluated with a reliable reference truth data prior to applications. And more literatures investigated the validation methods about the albedo validation in a flat or homogenous surface. However, the albedo performance over rugged terrain is still unknow due to the validation method limited. A multi-validation strategy is implemented to give a comprehensive albedo validation, which will involve the high resolution albedo processing, high resolution albedo validation based on in situ albedo, and the method to upscale the high resolution albedo to a coarse scale albedo. Among them, the high resolution albedo generation and the upscale method is the core step for the coarse scale albedo validation. In this paper, the high resolution albedo is generated by Angular Bin algorithm. And a albedo upscale method over rugged terrain is developed to obtain the coarse scale albedo truth. The in situ albedo located 40 sites in mountain area are selected globally to validate the high resolution albedo, and then upscaled to the coarse scale albedo by the upscale method. This paper takes MODIS and GLASS albedo product as a example, and the prelimarily results show the RMSE of MODIS and GLASS albedo product over rugged terrain are 0.047 and 0.057, respectively under the RMSE with 0.036 of high resolution albedo.

Dynamic Time Warping compared to established methods for validation of musculoskeletal models.

PubMed

Gaspar, Martin; Welke, Bastian; Seehaus, Frank; Hurschler, Christof; Schwarze, Michael

2017-04-11

By means of Multi-Body musculoskeletal simulation, important variables such as internal joint forces and moments can be estimated which cannot be measured directly. Validation can ensued by qualitative or by quantitative methods. Especially when comparing time-dependent signals, many methods do not perform well and validation is often limited to qualitative approaches. The aim of the present study was to investigate the capabilities of the Dynamic Time Warping (DTW) algorithm for comparing time series, which can quantify phase as well as amplitude errors. We contrast the sensitivity of DTW with other established metrics: the Pearson correlation coefficient, cross-correlation, the metric according to Geers, RMSE and normalized RMSE. This study is based on two data sets, where one data set represents direct validation and the other represents indirect validation. Direct validation was performed in the context of clinical gait-analysis on trans-femoral amputees fitted with a 6 component force-moment sensor. Measured forces and moments from amputees' socket-prosthesis are compared to simulated forces and moments. Indirect validation was performed in the context of surface EMG measurements on a cohort of healthy subjects with measurements taken of seven muscles of the leg, which were compared to simulated muscle activations. Regarding direct validation, a positive linear relation between results of RMSE and nRMSE to DTW can be seen. For indirect validation, a negative linear relation exists between Pearson correlation and cross-correlation. We propose the DTW algorithm for use in both direct and indirect quantitative validation as it correlates well with methods that are most suitable for one of the tasks. However, in DV it should be used together with methods resulting in a dimensional error value, in order to be able to interpret results more comprehensible. Copyright © 2017 Elsevier Ltd. All rights reserved.

Validation of a pulsed electric field process to pasteurize strawberry puree

USDA-ARS?s Scientific Manuscript database

An inexpensive data acquisition method was developed to validate the exact number and shape of the pulses applied during pulsed electric fields (PEF) processing. The novel validation method was evaluated in conjunction with developing a pasteurization PEF process for strawberry puree. Both buffered...

Use of the Method of Triads in the Validation of Sodium and Potassium Intake in the Brazilian Longitudinal Study of Adult Health (ELSA-Brasil).

PubMed

Pereira, Taísa Sabrina Silva; Cade, Nágela Valadão; Mill, José Geraldo; Sichieri, Rosely; Molina, Maria Del Carmen Bisi

2016-01-01

Biomarkers are a good choice to be used in the validation of food frequency questionnaire due to the independence of their random errors. To assess the validity of the potassium and sodium intake estimated using the Food Frequency Questionnaire ELSA-Brasil. A subsample of participants in the ELSA-Brasil cohort was included in this study in 2009. Sodium and potassium intake were estimated using three methods: Semi-quantitative food frequency questionnaire, 12-hour nocturnal urinary excretion and three 24-hour food records. Correlation coefficients were calculated between the methods, and the validity coefficient was calculated using the method of triads. The 95% confidence intervals for the validity coefficient were estimated using bootstrap sampling. Exact and adjacent agreement and disagreement of the estimated sodium and potassium intake quintiles were compared among three methods. The sample consisted of 246 participants, aged 53±8 years, 52% of women. Validity coefficient for sodium were considered weak (рfood frequency questionnaire actual intake = 0.37 and рbiomarker actual intake = 0.21) and moderate (рfood records actual intake 0.56). The validity coefficient were higher for potassium (рfood frequency questionnaire actual intake = 0.60; рbiomarker actual intake = 0.42; рfood records actual intake = 0.79). Conclusions: The Food Frequency Questionnaire ELSA-Brasil showed good validity in estimating potassium intake in epidemiological studies. For sodium validity was weak, likely due to the non-quantification of the added salt to prepared food.

International Harmonization and Cooperation in the Validation of Alternative Methods.

PubMed

Barroso, João; Ahn, Il Young; Caldeira, Cristiane; Carmichael, Paul L; Casey, Warren; Coecke, Sandra; Curren, Rodger; Desprez, Bertrand; Eskes, Chantra; Griesinger, Claudius; Guo, Jiabin; Hill, Erin; Roi, Annett Janusch; Kojima, Hajime; Li, Jin; Lim, Chae Hyung; Moura, Wlamir; Nishikawa, Akiyoshi; Park, HyeKyung; Peng, Shuangqing; Presgrave, Octavio; Singer, Tim; Sohn, Soo Jung; Westmoreland, Carl; Whelan, Maurice; Yang, Xingfen; Yang, Ying; Zuang, Valérie

The development and validation of scientific alternatives to animal testing is important not only from an ethical perspective (implementation of 3Rs), but also to improve safety assessment decision making with the use of mechanistic information of higher relevance to humans. To be effective in these efforts, it is however imperative that validation centres, industry, regulatory bodies, academia and other interested parties ensure a strong international cooperation, cross-sector collaboration and intense communication in the design, execution, and peer review of validation studies. Such an approach is critical to achieve harmonized and more transparent approaches to method validation, peer-review and recommendation, which will ultimately expedite the international acceptance of valid alternative methods or strategies by regulatory authorities and their implementation and use by stakeholders. It also allows achieving greater efficiency and effectiveness by avoiding duplication of effort and leveraging limited resources. In view of achieving these goals, the International Cooperation on Alternative Test Methods (ICATM) was established in 2009 by validation centres from Europe, USA, Canada and Japan. ICATM was later joined by Korea in 2011 and currently also counts with Brazil and China as observers. This chapter describes the existing differences across world regions and major efforts carried out for achieving consistent international cooperation and harmonization in the validation and adoption of alternative approaches to animal testing.

Determination of Ethanol in Kombucha Products: Single-Laboratory Validation, First Action 2016.12.

PubMed

Ebersole, Blake; Liu, Ying; Schmidt, Rich; Eckert, Matt; Brown, Paula N

2017-05-01

Kombucha is a fermented nonalcoholic beverage that has drawn government attention due to the possible presence of excess ethanol (≥0.5% alcohol by volume; ABV). A validated method that provides better precision and accuracy for measuring ethanol levels in kombucha is urgently needed by the kombucha industry. The current study validated a method for determining ethanol content in commercial kombucha products. The ethanol content in kombucha was measured using headspace GC with flame ionization detection. An ethanol standard curve ranging from 0.05 to 5.09% ABV was used, with correlation coefficients greater than 99.9%. The method detection limit was 0.003% ABV and the LOQ was 0.01% ABV. The RSDr ranged from 1.62 to 2.21% and the Horwitz ratio ranged from 0.4 to 0.6. The average accuracy of the method was 98.2%. This method was validated following the guidelines for single-laboratory validation by AOAC INTERNATIONAL and meets the requirements set by AOAC SMPR 2016.001, "Standard Method Performance Requirements for Determination of Ethanol in Kombucha."

DEVELOPMENT OF GUIDELINES FOR CALIBRATING, VALIDATING, AND EVALUATING HYDROLOGIC AND WATER QUALITY MODELS: ASABE ENGINEERING PRACTICE 621

USDA-ARS?s Scientific Manuscript database

Information to support application of hydrologic and water quality (H/WQ) models abounds, yet modelers commonly use arbitrary, ad hoc methods to conduct, document, and report model calibration, validation, and evaluation. Consistent methods are needed to improve model calibration, validation, and e...

A Framework for Mixing Methods in Quantitative Measurement Development, Validation, and Revision: A Case Study

ERIC Educational Resources Information Center

Luyt, Russell

2012-01-01

A framework for quantitative measurement development, validation, and revision that incorporates both qualitative and quantitative methods is introduced. It extends and adapts Adcock and Collier's work, and thus, facilitates understanding of quantitative measurement development, validation, and revision as an integrated and cyclical set of…

Evaluation of Criterion Validity for Scales with Congeneric Measures

ERIC Educational Resources Information Center

Raykov, Tenko

2007-01-01

A method for estimating criterion validity of scales with homogeneous components is outlined. It accomplishes point and interval estimation of interrelationship indices between composite scores and criterion variables and is useful for testing hypotheses about criterion validity of measurement instruments. The method can also be used with missing…

A Psychometric Study of the Bayley Scales of Infant and Toddler Development in Persian Language Children

PubMed Central

AZARI, Nadia; SOLEIMANI, Farin; VAMEGHI, Roshanak; SAJEDI, Firoozeh; SHAHSHAHANI, Soheila; KARIMI, Hossein; KRASKIAN, Adis; SHAHROKHI, Amin; TEYMOURI, Robab; GHARIB, Masoud

2017-01-01

Objective Bayley Scales of infant & toddler development is a well-known diagnostic developmental assessment tool for children aged 1–42 months. Our aim was investigating the validity & reliability of this scale in Persian speaking children. Materials & Methods The method was descriptive-analytic. Translation- back translation and cultural adaptation was done. Content & face validity of translated scale was determined by experts’ opinions. Overall, 403 children aged 1 to 42 months were recruited from health centers of Tehran, during years of 2013-2014 for developmental assessment in cognitive, communicative (receptive & expressive) and motor (fine & gross) domains. Reliability of scale was calculated through three methods; internal consistency using Cronbach’s alpha coefficient, test-retest and interrater methods. Construct validity was calculated using factor analysis and comparison of the mean scores methods. Results Cultural and linguistic changes were made in items of all domains especially on communication subscale. Content and face validity of the test were approved by experts’ opinions. Cronbach’s alpha coefficient was above 0.74 in all domains. Pearson correlation coefficient in various domains, were ≥ 0.982 in test retest method, and ≥0.993 in inter-rater method. Construct validity of the test was approved by factor analysis. Moreover, the mean scores for the different age groups were compared and statistically significant differences were observed between mean scores of different age groups, that confirms validity of the test. Conclusion The Bayley Scales of Infant and Toddler Development is a valid and reliable tool for child developmental assessment in Persian language children. PMID:28277556

Validity of three clinical performance assessments of internal medicine clerks.

PubMed

Hull, A L; Hodder, S; Berger, B; Ginsberg, D; Lindheim, N; Quan, J; Kleinhenz, M E

1995-06-01

To analyze the construct validity of three methods to assess the clinical performances of internal medicine clerks. A multitrait-multimethod (MTMM) study was conducted at the Case Western Reserve University School of Medicine to determine the convergent and divergent validity of a clinical evaluation form (CEF) completed by faculty and residents, an objective structured clinical examination (OSCE), and the medicine subject test of the National Board of Medical Examiners. Three traits were involved in the analysis: clinical skills, knowledge, and personal characteristics. A correlation matrix was computed for 410 third-year students who completed the clerkship between August 1988 and July 1991. There was a significant (p < .01) convergence of the four correlations that assessed the same traits by using different methods. However, the four convergent correlations were of moderate magnitude (ranging from .29 to .47). Divergent validity was assessed by comparing the magnitudes of the convergence correlations with the magnitudes of correlations among unrelated assessments (i.e., different traits by different methods). Seven of nine possible coefficients were smaller than the convergent coefficients, suggesting evidence of divergent validity. A significant CEF method effect was identified. There was convergent validity and some evidence of divergent validity with a significant method effect. The findings were similar for correlations corrected for attenuation. Four conclusions were reached: (1) the reliability of the OSCE must be improved, (2) the CEF ratings must be redesigned to further discriminate among the specific traits assessed, (3) additional methods to assess personal characteristics must be instituted, and (4) several assessment methods should be used to evaluate individual student performances.

Empirical Performance of Cross-Validation With Oracle Methods in a Genomics Context.

PubMed

Martinez, Josue G; Carroll, Raymond J; Müller, Samuel; Sampson, Joshua N; Chatterjee, Nilanjan

2011-11-01

When employing model selection methods with oracle properties such as the smoothly clipped absolute deviation (SCAD) and the Adaptive Lasso, it is typical to estimate the smoothing parameter by m-fold cross-validation, for example, m = 10. In problems where the true regression function is sparse and the signals large, such cross-validation typically works well. However, in regression modeling of genomic studies involving Single Nucleotide Polymorphisms (SNP), the true regression functions, while thought to be sparse, do not have large signals. We demonstrate empirically that in such problems, the number of selected variables using SCAD and the Adaptive Lasso, with 10-fold cross-validation, is a random variable that has considerable and surprising variation. Similar remarks apply to non-oracle methods such as the Lasso. Our study strongly questions the suitability of performing only a single run of m-fold cross-validation with any oracle method, and not just the SCAD and Adaptive Lasso.

Fault-tolerant clock synchronization validation methodology. [in computer systems

NASA Technical Reports Server (NTRS)

Butler, Ricky W.; Palumbo, Daniel L.; Johnson, Sally C.

1987-01-01

A validation method for the synchronization subsystem of a fault-tolerant computer system is presented. The high reliability requirement of flight-crucial systems precludes the use of most traditional validation methods. The method presented utilizes formal design proof to uncover design and coding errors and experimentation to validate the assumptions of the design proof. The experimental method is described and illustrated by validating the clock synchronization system of the Software Implemented Fault Tolerance computer. The design proof of the algorithm includes a theorem that defines the maximum skew between any two nonfaulty clocks in the system in terms of specific system parameters. Most of these parameters are deterministic. One crucial parameter is the upper bound on the clock read error, which is stochastic. The probability that this upper bound is exceeded is calculated from data obtained by the measurement of system parameters. This probability is then included in a detailed reliability analysis of the system.

Review of surface steam sterilization for validation purposes.

PubMed

van Doornmalen, Joost; Kopinga, Klaas

2008-03-01

Sterilization is an essential step in the process of producing sterile medical devices. To guarantee sterility, the process of sterilization must be validated. Because there is no direct way to measure sterility, the techniques applied to validate the sterilization process are based on statistical principles. Steam sterilization is the most frequently applied sterilization method worldwide and can be validated either by indicators (chemical or biological) or physical measurements. The steam sterilization conditions are described in the literature. Starting from these conditions, criteria for the validation of steam sterilization are derived and can be described in terms of physical parameters. Physical validation of steam sterilization appears to be an adequate and efficient validation method that could be considered as an alternative for indicator validation. Moreover, physical validation can be used for effective troubleshooting in steam sterilizing processes.

A systematic review and appraisal of methods of developing and validating lifestyle cardiovascular disease risk factors questionnaires.

PubMed

Nse, Odunaiya; Quinette, Louw; Okechukwu, Ogah

2015-09-01

Well developed and validated lifestyle cardiovascular disease (CVD) risk factors questionnaires is the key to obtaining accurate information to enable planning of CVD prevention program which is a necessity in developing countries. We conducted this review to assess methods and processes used for development and content validation of lifestyle CVD risk factors questionnaires and possibly develop an evidence based guideline for development and content validation of lifestyle CVD risk factors questionnaires. Relevant databases at the Stellenbosch University library were searched for studies conducted between 2008 and 2012, in English language and among humans. Using the following databases; pubmed, cinahl, psyc info and proquest. Search terms used were CVD risk factors, questionnaires, smoking, alcohol, physical activity and diet. Methods identified for development of lifestyle CVD risk factors were; review of literature either systematic or traditional, involvement of expert and /or target population using focus group discussion/interview, clinical experience of authors and deductive reasoning of authors. For validation, methods used were; the involvement of expert panel, the use of target population and factor analysis. Combination of methods produces questionnaires with good content validity and other psychometric properties which we consider good.

Analysis of Ethanolamines: Validation of Semi-Volatile Analysis by HPLC-MS/MS by EPA Method MS888

DOE Office of Scientific and Technical Information (OSTI.GOV)

Owens, J; Vu, A; Koester, C

The Environmental Protection Agency's (EPA) Region 5 Chicago Regional Laboratory (CRL) developed a method titled 'Analysis of Diethanolamine, Triethanolamine, n-Methyldiethanolamine, and n-Ethyldiethanolamine in Water by Single Reaction Monitoring Liquid Chromatography/Tandem Mass Spectrometry (LC/MS/MS): EPA Method MS888'. This draft standard operating procedure (SOP) was distributed to multiple EPA laboratories and to Lawrence Livermore National Laboratory, which was tasked to serve as a reference laboratory for EPA's Environmental Reference Laboratory Network (ERLN) and to develop and validate analytical procedures. The primary objective of this study was to validate and verify the analytical procedures described in 'EPA Method MS888' for analysis of themore » listed ethanolamines in aqueous samples. The gathered data from this validation study will be used to: (1) demonstrate analytical method performance; (2) generate quality control acceptance criteria; and (3) revise the SOP to provide a validated method that would be available for use during a homeland security event. The data contained in this report will be compiled, by EPA CRL, with data generated by other EPA Regional laboratories so that performance metrics of 'EPA Method MS888' can be determined.« less

A Psychometric Study of the Bayley Scales of Infant and Toddler Development in Persian Language Children.

PubMed

Azari, Nadia; Soleimani, Farin; Vameghi, Roshanak; Sajedi, Firoozeh; Shahshahani, Soheila; Karimi, Hossein; Kraskian, Adis; Shahrokhi, Amin; Teymouri, Robab; Gharib, Masoud

2017-01-01

Bayley Scales of infant & toddler development is a well-known diagnostic developmental assessment tool for children aged 1-42 months. Our aim was investigating the validity & reliability of this scale in Persian speaking children. The method was descriptive-analytic. Translation- back translation and cultural adaptation was done. Content & face validity of translated scale was determined by experts' opinions. Overall, 403 children aged 1 to 42 months were recruited from health centers of Tehran, during years of 2013-2014 for developmental assessment in cognitive, communicative (receptive & expressive) and motor (fine & gross) domains. Reliability of scale was calculated through three methods; internal consistency using Cronbach's alpha coefficient, test-retest and interrater methods. Construct validity was calculated using factor analysis and comparison of the mean scores methods. Cultural and linguistic changes were made in items of all domains especially on communication subscale. Content and face validity of the test were approved by experts' opinions. Cronbach's alpha coefficient was above 0.74 in all domains. Pearson correlation coefficient in various domains, were ≥ 0.982 in test retest method, and ≥0.993 in inter-rater method. Construct validity of the test was approved by factor analysis. Moreover, the mean scores for the different age groups were compared and statistically significant differences were observed between mean scores of different age groups, that confirms validity of the test. The Bayley Scales of Infant and Toddler Development is a valid and reliable tool for child developmental assessment in Persian language children.

A method for validation of finite element forming simulation on basis of a pointwise comparison of distance and curvature

NASA Astrophysics Data System (ADS)

Dörr, Dominik; Joppich, Tobias; Schirmaier, Fabian; Mosthaf, Tobias; Kärger, Luise; Henning, Frank

2016-10-01

Thermoforming of continuously fiber reinforced thermoplastics (CFRTP) is ideally suited to thin walled and complex shaped products. By means of forming simulation, an initial validation of the producibility of a specific geometry, an optimization of the forming process and the prediction of fiber-reorientation due to forming is possible. Nevertheless, applied methods need to be validated. Therefor a method is presented, which enables the calculation of error measures for the mismatch between simulation results and experimental tests, based on measurements with a conventional coordinate measuring device. As a quantitative measure, describing the curvature is provided, the presented method is also suitable for numerical or experimental sensitivity studies on wrinkling behavior. The applied methods for forming simulation, implemented in Abaqus explicit, are presented and applied to a generic geometry. The same geometry is tested experimentally and simulation and test results are compared by the proposed validation method.

Discriminant content validity: a quantitative methodology for assessing content of theory-based measures, with illustrative applications.

PubMed

Johnston, Marie; Dixon, Diane; Hart, Jo; Glidewell, Liz; Schröder, Carin; Pollard, Beth

2014-05-01

In studies involving theoretical constructs, it is important that measures have good content validity and that there is not contamination of measures by content from other constructs. While reliability and construct validity are routinely reported, to date, there has not been a satisfactory, transparent, and systematic method of assessing and reporting content validity. In this paper, we describe a methodology of discriminant content validity (DCV) and illustrate its application in three studies. Discriminant content validity involves six steps: construct definition, item selection, judge identification, judgement format, single-sample test of content validity, and assessment of discriminant items. In three studies, these steps were applied to a measure of illness perceptions (IPQ-R) and control cognitions. The IPQ-R performed well with most items being purely related to their target construct, although timeline and consequences had small problems. By contrast, the study of control cognitions identified problems in measuring constructs independently. In the final study, direct estimation response formats for theory of planned behaviour constructs were found to have as good DCV as Likert format. The DCV method allowed quantitative assessment of each item and can therefore inform the content validity of the measures assessed. The methods can be applied to assess content validity before or after collecting data to select the appropriate items to measure theoretical constructs. Further, the data reported for each item in Appendix S1 can be used in item or measure selection. Statement of contribution What is already known on this subject? There are agreed methods of assessing and reporting construct validity of measures of theoretical constructs, but not their content validity. Content validity is rarely reported in a systematic and transparent manner. What does this study add? The paper proposes discriminant content validity (DCV), a systematic and transparent method of assessing and reporting whether items assess the intended theoretical construct and only that construct. In three studies, DCV was applied to measures of illness perceptions, control cognitions, and theory of planned behaviour response formats. Appendix S1 gives content validity indices for each item of each questionnaire investigated. Discriminant content validity is ideally applied while the measure is being developed, before using to measure the construct(s), but can also be applied after using a measure. © 2014 The British Psychological Society.

A brief measure of attitudes toward mixed methods research in psychology.

PubMed

Roberts, Lynne D; Povee, Kate

2014-01-01

The adoption of mixed methods research in psychology has trailed behind other social science disciplines. Teaching psychology students, academics, and practitioners about mixed methodologies may increase the use of mixed methods within the discipline. However, tailoring and evaluating education and training in mixed methodologies requires an understanding of, and way of measuring, attitudes toward mixed methods research in psychology. To date, no such measure exists. In this article we present the development and initial validation of a new measure: Attitudes toward Mixed Methods Research in Psychology. A pool of 42 items developed from previous qualitative research on attitudes toward mixed methods research along with validation measures was administered via an online survey to a convenience sample of 274 psychology students, academics and psychologists. Principal axis factoring with varimax rotation on a subset of the sample produced a four-factor, 12-item solution. Confirmatory factor analysis on a separate subset of the sample indicated that a higher order four factor model provided the best fit to the data. The four factors; 'Limited Exposure,' '(in)Compatibility,' 'Validity,' and 'Tokenistic Qualitative Component'; each have acceptable internal reliability. Known groups validity analyses based on preferred research orientation and self-rated mixed methods research skills, and convergent and divergent validity analyses based on measures of attitudes toward psychology as a science and scientist and practitioner orientation, provide initial validation of the measure. This brief, internally reliable measure can be used in assessing attitudes toward mixed methods research in psychology, measuring change in attitudes as part of the evaluation of mixed methods education, and in larger research programs.

Verification and Validation Studies for the LAVA CFD Solver

NASA Technical Reports Server (NTRS)

Moini-Yekta, Shayan; Barad, Michael F; Sozer, Emre; Brehm, Christoph; Housman, Jeffrey A.; Kiris, Cetin C.

2013-01-01

The verification and validation of the Launch Ascent and Vehicle Aerodynamics (LAVA) computational fluid dynamics (CFD) solver is presented. A modern strategy for verification and validation is described incorporating verification tests, validation benchmarks, continuous integration and version control methods for automated testing in a collaborative development environment. The purpose of the approach is to integrate the verification and validation process into the development of the solver and improve productivity. This paper uses the Method of Manufactured Solutions (MMS) for the verification of 2D Euler equations, 3D Navier-Stokes equations as well as turbulence models. A method for systematic refinement of unstructured grids is also presented. Verification using inviscid vortex propagation and flow over a flat plate is highlighted. Simulation results using laminar and turbulent flow past a NACA 0012 airfoil and ONERA M6 wing are validated against experimental and numerical data.

An intercomparison of a large ensemble of statistical downscaling methods for Europe: Overall results from the VALUE perfect predictor cross-validation experiment

NASA Astrophysics Data System (ADS)

Gutiérrez, Jose Manuel; Maraun, Douglas; Widmann, Martin; Huth, Radan; Hertig, Elke; Benestad, Rasmus; Roessler, Ole; Wibig, Joanna; Wilcke, Renate; Kotlarski, Sven

2016-04-01

VALUE is an open European network to validate and compare downscaling methods for climate change research (http://www.value-cost.eu). A key deliverable of VALUE is the development of a systematic validation framework to enable the assessment and comparison of both dynamical and statistical downscaling methods. This framework is based on a user-focused validation tree, guiding the selection of relevant validation indices and performance measures for different aspects of the validation (marginal, temporal, spatial, multi-variable). Moreover, several experiments have been designed to isolate specific points in the downscaling procedure where problems may occur (assessment of intrinsic performance, effect of errors inherited from the global models, effect of non-stationarity, etc.). The list of downscaling experiments includes 1) cross-validation with perfect predictors, 2) GCM predictors -aligned with EURO-CORDEX experiment- and 3) pseudo reality predictors (see Maraun et al. 2015, Earth's Future, 3, doi:10.1002/2014EF000259, for more details). The results of these experiments are gathered, validated and publicly distributed through the VALUE validation portal, allowing for a comprehensive community-open downscaling intercomparison study. In this contribution we describe the overall results from Experiment 1), consisting of a European wide 5-fold cross-validation (with consecutive 6-year periods from 1979 to 2008) using predictors from ERA-Interim to downscale precipitation and temperatures (minimum and maximum) over a set of 86 ECA&D stations representative of the main geographical and climatic regions in Europe. As a result of the open call for contribution to this experiment (closed in Dec. 2015), over 40 methods representative of the main approaches (MOS and Perfect Prognosis, PP) and techniques (linear scaling, quantile mapping, analogs, weather typing, linear and generalized regression, weather generators, etc.) were submitted, including information both data (downscaled values) and metadata (characterizing different aspects of the downscaling methods). This constitutes the largest and most comprehensive to date intercomparison of statistical downscaling methods. Here, we present an overall validation, analyzing marginal and temporal aspects to assess the intrinsic performance and added value of statistical downscaling methods at both annual and seasonal levels. This validation takes into account the different properties/limitations of different approaches and techniques (as reported in the provided metadata) in order to perform a fair comparison. It is pointed out that this experiment alone is not sufficient to evaluate the limitations of (MOS) bias correction techniques. Moreover, it also does not fully validate PP since we don't learn whether we have the right predictors and whether the PP assumption is valid. These problems will be analyzed in the subsequent community-open VALUE experiments 2) and 3), which will be open for participation along the present year.

Determination of Phosphorus and Potassium in Commercial Inorganic Fertilizers by Inductively Coupled Plasma-Optical Emission Spectrometry: Single-Laboratory Validation, First Action 2015.18.

PubMed

Thiex, Nancy J

2016-07-01

A previously validated method for the determination of both citrate-EDTA-soluble P and K and acid-soluble P and K in commercial inorganic fertilizers by inductively coupled plasma-optical emission spectrometry was submitted to the expert review panel (ERP) for fertilizers for consideration of First Action Official Method(SM) status. The ERP evaluated the single-laboratory validation results and recommended the method for First Action Official Method status and provided recommendations for achieving Final Action. Validation materials ranging from 4.4 to 52.4% P2O5 (1.7-22.7% P) and 3-62% K2O (2.5-51.1% K) were used for the validation. Recoveries from validation materials for citrate-soluble P and K ranged from 99.3 to 124.9% P and from 98.4 to 100.7% K. Recoveries from validation materials for acid-soluble "total" P and K ranged from 95.53 to 99.40% P and from 98.36 to 107.28% K. Values of r for citrate-soluble P and K, expressed as RSD, ranged from 0.28 to 1.30% for P and from 0.41 to 1.52% for K. Values of r for total P and K, expressed as RSD, ranged from 0.71 to 1.13% for P and from 0.39 to 1.18% for K. Based on the validation data, the ERP recommended the method (with alternatives for the citrate-soluble and the acid-soluble extractions) for First Action Official Method status and provided recommendations for achieving Final Action status.

DBS-LC-MS/MS assay for caffeine: validation and neonatal application.

PubMed

Bruschettini, Matteo; Barco, Sebastiano; Romantsik, Olga; Risso, Francesco; Gennai, Iulian; Chinea, Benito; Ramenghi, Luca A; Tripodi, Gino; Cangemi, Giuliana

2016-09-01

DBS might be an appropriate microsampling technique for therapeutic drug monitoring of caffeine in infants. Nevertheless, its application presents several issues that still limit its use. This paper describes a validated DBS-LC-MS/MS method for caffeine. The results of the method validation showed an hematocrit dependence. In the analysis of 96 paired plasma and DBS clinical samples, caffeine levels measured in DBS were statistically significantly lower than in plasma but the observed differences were independent from hematocrit. These results clearly showed the need for extensive validation with real-life samples for DBS-based methods. DBS-LC-MS/MS can be considered to be a good alternative to traditional methods for therapeutic drug monitoring or PK studies in preterm infants.

Effective data validation of high-frequency data: time-point-, time-interval-, and trend-based methods.

PubMed

Horn, W; Miksch, S; Egghart, G; Popow, C; Paky, F

1997-09-01

Real-time systems for monitoring and therapy planning, which receive their data from on-line monitoring equipment and computer-based patient records, require reliable data. Data validation has to utilize and combine a set of fast methods to detect, eliminate, and repair faulty data, which may lead to life-threatening conclusions. The strength of data validation results from the combination of numerical and knowledge-based methods applied to both continuously-assessed high-frequency data and discontinuously-assessed data. Dealing with high-frequency data, examining single measurements is not sufficient. It is essential to take into account the behavior of parameters over time. We present time-point-, time-interval-, and trend-based methods for validation and repair. These are complemented by time-independent methods for determining an overall reliability of measurements. The data validation benefits from the temporal data-abstraction process, which provides automatically derived qualitative values and patterns. The temporal abstraction is oriented on a context-sensitive and expectation-guided principle. Additional knowledge derived from domain experts forms an essential part for all of these methods. The methods are applied in the field of artificial ventilation of newborn infants. Examples from the real-time monitoring and therapy-planning system VIE-VENT illustrate the usefulness and effectiveness of the methods.

An integrated bioanalytical method development and validation approach: case studies.

PubMed

Xue, Y-J; Melo, Brian; Vallejo, Martha; Zhao, Yuwen; Tang, Lina; Chen, Yuan-Shek; Keller, Karin M

2012-10-01

We proposed an integrated bioanalytical method development and validation approach: (1) method screening based on analyte's physicochemical properties and metabolism information to determine the most appropriate extraction/analysis conditions; (2) preliminary stability evaluation using both quality control and incurred samples to establish sample collection, storage and processing conditions; (3) mock validation to examine method accuracy and precision and incurred sample reproducibility; and (4) method validation to confirm the results obtained during method development. This integrated approach was applied to the determination of compound I in rat plasma and compound II in rat and dog plasma. The effectiveness of the approach was demonstrated by the superior quality of three method validations: (1) a zero run failure rate; (2) >93% of quality control results within 10% of nominal values; and (3) 99% incurred sample within 9.2% of the original values. In addition, rat and dog plasma methods for compound II were successfully applied to analyze more than 900 plasma samples obtained from Investigational New Drug (IND) toxicology studies in rats and dogs with near perfect results: (1) a zero run failure rate; (2) excellent accuracy and precision for standards and quality controls; and (3) 98% incurred samples within 15% of the original values. Copyright © 2011 John Wiley & Sons, Ltd.

Experimental design methodologies in the optimization of chiral CE or CEC separations: an overview.

PubMed

Dejaegher, Bieke; Mangelings, Debby; Vander Heyden, Yvan

2013-01-01

In this chapter, an overview of experimental designs to develop chiral capillary electrophoresis (CE) and capillary electrochromatographic (CEC) methods is presented. Method development is generally divided into technique selection, method optimization, and method validation. In the method optimization part, often two phases can be distinguished, i.e., a screening and an optimization phase. In method validation, the method is evaluated on its fit for purpose. A validation item, also applying experimental designs, is robustness testing. In the screening phase and in robustness testing, screening designs are applied. During the optimization phase, response surface designs are used. The different design types and their application steps are discussed in this chapter and illustrated by examples of chiral CE and CEC methods.

Teaching method validation in the clinical laboratory science curriculum.

PubMed

Moon, Tara C; Legrys, Vicky A

2008-01-01

With the Clinical Laboratory Improvement Amendment's (CLIA) final rule, the ability of the Clinical Laboratory Scientist (CLS) to perform method validation has become increasingly important. Knowledge of the statistical methods and procedures used in method validation is imperative for clinical laboratory scientists. However, incorporating these concepts in a CLS curriculum can be challenging, especially at a time of limited resources. This paper provides an outline of one approach to addressing these topics in lecture courses and integrating them in the student laboratory and the clinical practicum for direct application.

Applying the Mixed Methods Instrument Development and Construct Validation Process: the Transformative Experience Questionnaire

ERIC Educational Resources Information Center

Koskey, Kristin L. K.; Sondergeld, Toni A.; Stewart, Victoria C.; Pugh, Kevin J.

2018-01-01

Onwuegbuzie and colleagues proposed the Instrument Development and Construct Validation (IDCV) process as a mixed methods framework for creating and validating measures. Examples applying IDCV are lacking. We provide an illustrative case integrating the Rasch model and cognitive interviews applied to the development of the Transformative…

Validity of a digital diet estimation method for use with preschool children

USDA-ARS?s Scientific Manuscript database

The validity of using the Remote Food Photography Method (RFPM) for measuring food intake of minority preschool children's intake is not well documented. The aim of the study was to determine the validity of intake estimations made by human raters using the RFPM compared with those obtained by weigh...

Validating Accelerometry and Skinfold Measures in Youth with Down Syndrome

ERIC Educational Resources Information Center

Esposito, Phil Michael

2012-01-01

Current methods for measuring quantity and intensity of physical activity based on accelerometer output have been studied and validated in youth. These methods have been applied to youth with Down syndrome (DS) with no empirical research done to validate these measures. Similarly, individuals with DS have unique body proportions not represented by…

Validation of "Teaching and Learning Guiding Principles Instrument" for Malaysian Higher Learning Institutions

ERIC Educational Resources Information Center

Rahman, Nurulhuda Abd; Masuwai, Azwani; Tajudin, Nor'ain Mohd; Tek, Ong Eng; Adnan, Mazlini

2016-01-01

Purpose: This study was aimed at establishing, through the validation of the "Teaching and Learning Guiding Principles Instrument" (TLGPI), the validity and reliability of the underlying factor structure of the Teaching and Learning Guiding Principles (TLGP) generated by a previous study. Method: A survey method was used to collect data…

Double Cross-Validation in Multiple Regression: A Method of Estimating the Stability of Results.

ERIC Educational Resources Information Center

Rowell, R. Kevin

In multiple regression analysis, where resulting predictive equation effectiveness is subject to shrinkage, it is especially important to evaluate result replicability. Double cross-validation is an empirical method by which an estimate of invariance or stability can be obtained from research data. A procedure for double cross-validation is…

Analyzing the Validity of the Adult-Adolescent Parenting Inventory for Low-Income Populations

ERIC Educational Resources Information Center

Lawson, Michael A.; Alameda-Lawson, Tania; Byrnes, Edward

2017-01-01

Objectives: The purpose of this study was to examine the construct and predictive validity of the Adult-Adolescent Parenting Inventory (AAPI-2). Methods: The validity of the AAPI-2 was evaluated using multiple statistical methods, including exploratory factor analysis, confirmatory factor analysis, and latent class analysis. These analyses were…

Evolving the Principles and Practice of Validation for New Alternative Approaches to Toxicity Testing.

PubMed

Whelan, Maurice; Eskes, Chantra

Validation is essential for the translation of newly developed alternative approaches to animal testing into tools and solutions suitable for regulatory applications. Formal approaches to validation have emerged over the past 20 years or so and although they have helped greatly to progress the field, it is essential that the principles and practice underpinning validation continue to evolve to keep pace with scientific progress. The modular approach to validation should be exploited to encourage more innovation and flexibility in study design and to increase efficiency in filling data gaps. With the focus now on integrated approaches to testing and assessment that are based on toxicological knowledge captured as adverse outcome pathways, and which incorporate the latest in vitro and computational methods, validation needs to adapt to ensure it adds value rather than hinders progress. Validation needs to be pursued both at the method level, to characterise the performance of in vitro methods in relation their ability to detect any association of a chemical with a particular pathway or key toxicological event, and at the methodological level, to assess how integrated approaches can predict toxicological endpoints relevant for regulatory decision making. To facilitate this, more emphasis needs to be given to the development of performance standards that can be applied to classes of methods and integrated approaches that provide similar information. Moreover, the challenge of selecting the right reference chemicals to support validation needs to be addressed more systematically, consistently and in a manner that better reflects the state of the science. Above all however, validation requires true partnership between the development and user communities of alternative methods and the appropriate investment of resources.

Standardization, evaluation and early-phase method validation of an analytical scheme for batch-consistency N-glycosylation analysis of recombinant produced glycoproteins.

PubMed

Zietze, Stefan; Müller, Rainer H; Brecht, René

2008-03-01

In order to set up a batch-to-batch-consistency analytical scheme for N-glycosylation analysis, several sample preparation steps including enzyme digestions and fluorophore labelling and two HPLC-methods were established. The whole method scheme was standardized, evaluated and validated according to the requirements on analytical testing in early clinical drug development by usage of a recombinant produced reference glycoprotein (RGP). The standardization of the methods was performed by clearly defined standard operation procedures. During evaluation of the methods, the major interest was in the loss determination of oligosaccharides within the analytical scheme. Validation of the methods was performed with respect to specificity, linearity, repeatability, LOD and LOQ. Due to the fact that reference N-glycan standards were not available, a statistical approach was chosen to derive accuracy from the linearity data. After finishing the validation procedure, defined limits for method variability could be calculated and differences observed in consistency analysis could be separated into significant and incidental ones.

The predictive validity of a situational judgement test, a clinical problem solving test and the core medical training selection methods for performance in specialty training .

PubMed

Patterson, Fiona; Lopes, Safiatu; Harding, Stephen; Vaux, Emma; Berkin, Liz; Black, David

2017-02-01

The aim of this study was to follow up a sample of physicians who began core medical training (CMT) in 2009. This paper examines the long-term validity of CMT and GP selection methods in predicting performance in the Membership of Royal College of Physicians (MRCP(UK)) examinations. We performed a longitudinal study, examining the extent to which the GP and CMT selection methods (T1) predict performance in the MRCP(UK) examinations (T2). A total of 2,569 applicants from 2008-09 who completed CMT and GP selection methods were included in the study. Looking at MRCP(UK) part 1, part 2 written and PACES scores, both CMT and GP selection methods show evidence of predictive validity for the outcome variables, and hierarchical regressions show the GP methods add significant value to the CMT selection process. CMT selection methods predict performance in important outcomes and have good evidence of validity; the GP methods may have an additional role alongside the CMT selection methods. © Royal College of Physicians 2017. All rights reserved.

Investigation of Content and Face Validity and Reliability of Sociocultural Attitude towards Appearance Questionnaire-3 (SATAQ-3) among Female Adolescents

PubMed Central

Mousazadeh, Somayeh; Rakhshan, Mahnaz; Mohammadi, Fateme

2017-01-01

Objective: This study aimed to determine the psychometric properties of sociocultural attitude towards appearance questionnaire in female adolescents. Method: This was a methodological study. The English version of the questionnaire was translated into Persian, using forward-backward method. Then the face validity, content validity and reliability were checked. To ensure face validity, the questionnaire was given to 25 female adolescents, a psychologist and three nurses, who were required to evaluate the items with respect to problems, ambiguity, relativity, proper terms and grammar, and understandability. For content validity, 15 experts in psychology and nursing, who met the inclusion criteria, were required. They were asked to assess the qualitative of content validity. To determine the quantitative content validity, content validity index and content validity ratio were calculated. At the end, internal consistency of the items was assessed, using Cronbach’s alpha method. Results: According to the expert judgments, content validity ratio was 0.81 and content validity index was 0.91. Besides, the reliability of the questionnaire was confirmed with Cronbach’s alpha = 0.91, and physical and developmental areas showed the highest reliability indices. Conclusion: The aforementioned questionnaire could be used in researches to assess female adolescents’ self-concept. This can be a stepping-stone towards identification of problems and improvement of adolescents’ body image. PMID:28496497

A method for sensitivity analysis to assess the effects of measurement error in multiple exposure variables using external validation data.

PubMed

Agogo, George O; van der Voet, Hilko; van 't Veer, Pieter; Ferrari, Pietro; Muller, David C; Sánchez-Cantalejo, Emilio; Bamia, Christina; Braaten, Tonje; Knüppel, Sven; Johansson, Ingegerd; van Eeuwijk, Fred A; Boshuizen, Hendriek C

2016-10-13

Measurement error in self-reported dietary intakes is known to bias the association between dietary intake and a health outcome of interest such as risk of a disease. The association can be distorted further by mismeasured confounders, leading to invalid results and conclusions. It is, however, difficult to adjust for the bias in the association when there is no internal validation data. We proposed a method to adjust for the bias in the diet-disease association (hereafter, association), due to measurement error in dietary intake and a mismeasured confounder, when there is no internal validation data. The method combines prior information on the validity of the self-report instrument with the observed data to adjust for the bias in the association. We compared the proposed method with the method that ignores the confounder effect, and with the method that ignores measurement errors completely. We assessed the sensitivity of the estimates to various magnitudes of measurement error, error correlations and uncertainty in the literature-reported validation data. We applied the methods to fruits and vegetables (FV) intakes, cigarette smoking (confounder) and all-cause mortality data from the European Prospective Investigation into Cancer and Nutrition study. Using the proposed method resulted in about four times increase in the strength of association between FV intake and mortality. For weakly correlated errors, measurement error in the confounder minimally affected the hazard ratio estimate for FV intake. The effect was more pronounced for strong error correlations. The proposed method permits sensitivity analysis on measurement error structures and accounts for uncertainties in the reported validity coefficients. The method is useful in assessing the direction and quantifying the magnitude of bias in the association due to measurement errors in the confounders.

Content Validity of National Post Marriage Educational Program Using Mixed Methods

PubMed Central

MOHAJER RAHBARI, Masoumeh; SHARIATI, Mohammad; KERAMAT, Afsaneh; YUNESIAN, Masoud; ESLAMI, Mohammad; MOUSAVI, Seyed Abbas; MONTAZERI, Ali

2015-01-01

Background: Although the validity of content of program is mostly conducted with qualitative methods, this study used both qualitative and quantitative methods for the validation of content of post marriage training program provided for newly married couples. Content validity is a preliminary step of obtaining authorization required to install the program in country's health care system. Methods: This mixed methodological content validation study carried out in four steps with forming three expert panels. Altogether 24 expert panelists were involved in 3 qualitative and quantitative panels; 6 in the first item development one; 12 in the reduction kind, 4 of them were common with the first panel, and 10 executive experts in the last one organized to evaluate psychometric properties of CVR and CVI and Face validity of 57 educational objectives. Results: The raw data of post marriage program had been written by professional experts of Ministry of Health, using qualitative expert panel, the content was more developed by generating 3 topics and refining one topic and its respective content. In the second panel, totally six other objectives were deleted, three for being out of agreement cut of point and three on experts' consensus. The validity of all items was above 0.8 and their content validity indices (0.8–1) were completely appropriate in quantitative assessment. Conclusion: This study provided a good evidence for validation and accreditation of national post marriage program planned for newly married couples in health centers of the country in the near future. PMID:26056672

Development of an Itemwise Efficiency Scoring Method: Concurrent, Convergent, Discriminant, and Neuroimaging-Based Predictive Validity Assessed in a Large Community Sample

PubMed Central

Moore, Tyler M.; Reise, Steven P.; Roalf, David R.; Satterthwaite, Theodore D.; Davatzikos, Christos; Bilker, Warren B.; Port, Allison M.; Jackson, Chad T.; Ruparel, Kosha; Savitt, Adam P.; Baron, Robert B.; Gur, Raquel E.; Gur, Ruben C.

2016-01-01

Traditional “paper-and-pencil” testing is imprecise in measuring speed and hence limited in assessing performance efficiency, but computerized testing permits precision in measuring itemwise response time. We present a method of scoring performance efficiency (combining information from accuracy and speed) at the item level. Using a community sample of 9,498 youths age 8-21, we calculated item-level efficiency scores on four neurocognitive tests, and compared the concurrent, convergent, discriminant, and predictive validity of these scores to simple averaging of standardized speed and accuracy-summed scores. Concurrent validity was measured by the scores' abilities to distinguish men from women and their correlations with age; convergent and discriminant validity were measured by correlations with other scores inside and outside of their neurocognitive domains; predictive validity was measured by correlations with brain volume in regions associated with the specific neurocognitive abilities. Results provide support for the ability of itemwise efficiency scoring to detect signals as strong as those detected by standard efficiency scoring methods. We find no evidence of superior validity of the itemwise scores over traditional scores, but point out several advantages of the former. The itemwise efficiency scoring method shows promise as an alternative to standard efficiency scoring methods, with overall moderate support from tests of four different types of validity. This method allows the use of existing item analysis methods and provides the convenient ability to adjust the overall emphasis of accuracy versus speed in the efficiency score, thus adjusting the scoring to the real-world demands the test is aiming to fulfill. PMID:26866796

Best Practices in Stability Indicating Method Development and Validation for Non-clinical Dose Formulations.

PubMed

Henry, Teresa R; Penn, Lara D; Conerty, Jason R; Wright, Francesca E; Gorman, Gregory; Pack, Brian W

2016-11-01

Non-clinical dose formulations (also known as pre-clinical or GLP formulations) play a key role in early drug development. These formulations are used to introduce active pharmaceutical ingredients (APIs) into test organisms for both pharmacokinetic and toxicological studies. Since these studies are ultimately used to support dose and safety ranges in human studies, it is important to understand not only the concentration and PK/PD of the active ingredient but also to generate safety data for likely process impurities and degradation products of the active ingredient. As such, many in the industry have chosen to develop and validate methods which can accurately detect and quantify the active ingredient along with impurities and degradation products. Such methods often provide trendable results which are predictive of stability, thus leading to the name; stability indicating methods. This document provides an overview of best practices for those choosing to include development and validation of such methods as part of their non-clinical drug development program. This document is intended to support teams who are either new to stability indicating method development and validation or who are less familiar with the requirements of validation due to their position within the product development life cycle.

Analysis of Carbamate Pesticides: Validation of Semi-Volatile Analysis by HPLC-MS/MS by EPA Method MS666

DOE Office of Scientific and Technical Information (OSTI.GOV)

Owens, J; Koester, C

The Environmental Protection Agency's (EPA) Region 5 Chicago Regional Laboratory (CRL) developed a method for analysis of aldicarb, bromadiolone, carbofuran, oxamyl, and methomyl in water by high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS), titled Method EPA MS666. This draft standard operating procedure (SOP) was distributed to multiple EPA laboratories and to Lawrence Livermore National Laboratory, which was tasked to serve as a reference laboratory for EPA's Environmental Reference Laboratory Network (ERLN) and to develop and validate analytical procedures. The primary objective of this study was to validate and verify the analytical procedures described in MS666 for analysis of carbamatemore » pesticides in aqueous samples. The gathered data from this validation study will be used to: (1) demonstrate analytical method performance; (2) generate quality control acceptance criteria; and (3) revise the SOP to provide a validated method that would be available for use during a homeland security event. The data contained in this report will be compiled, by EPA CRL, with data generated by other EPA Regional laboratories so that performance metrics of Method EPA MS666 can be determined.« less

A Comparison of Three Methods for the Analysis of Skin Flap Viability: Reliability and Validity.

PubMed

Tim, Carla Roberta; Martignago, Cintia Cristina Santi; da Silva, Viviane Ribeiro; Dos Santos, Estefany Camila Bonfim; Vieira, Fabiana Nascimento; Parizotto, Nivaldo Antonio; Liebano, Richard Eloin

2018-05-01

Objective: Technological advances have provided new alternatives to the analysis of skin flap viability in animal models; however, the interrater validity and reliability of these techniques have yet to be analyzed. The present study aimed to evaluate the interrater validity and reliability of three different methods: weight of paper template (WPT), paper template area (PTA), and photographic analysis. Approach: Sixteen male Wistar rats had their cranially based dorsal skin flap elevated. On the seventh postoperative day, the viable tissue area and the necrotic area of the skin flap were recorded using the paper template method and photo image. The evaluation of the percentage of viable tissue was performed using three methods, simultaneously and independently by two raters. The analysis of interrater reliability and viability was performed using the intraclass correlation coefficient and Bland Altman Plot Analysis was used to visualize the presence or absence of systematic bias in the evaluations of data validity. Results: The results showed that interrater reliability for WPT, measurement of PTA, and photographic analysis were 0.995, 0.990, and 0.982, respectively. For data validity, a correlation >0.90 was observed for all comparisons made between the three methods. In addition, Bland Altman Plot Analysis showed agreement between the comparisons of the methods and the presence of systematic bias was not observed. Innovation: Digital methods are an excellent choice for assessing skin flap viability; moreover, they make data use and storage easier. Conclusion: Independently from the method used, the interrater reliability and validity proved to be excellent for the analysis of skin flaps' viability.

Global approach for the validation of an in-line Raman spectroscopic method to determine the API content in real-time during a hot-melt extrusion process.

PubMed

Netchacovitch, L; Thiry, J; De Bleye, C; Dumont, E; Cailletaud, J; Sacré, P-Y; Evrard, B; Hubert, Ph; Ziemons, E

2017-08-15

Since the Food and Drug Administration (FDA) published a guidance based on the Process Analytical Technology (PAT) approach, real-time analyses during manufacturing processes are in real expansion. In this study, in-line Raman spectroscopic analyses were performed during a Hot-Melt Extrusion (HME) process to determine the Active Pharmaceutical Ingredient (API) content in real-time. The method was validated based on a univariate and a multivariate approach and the analytical performances of the obtained models were compared. Moreover, on one hand, in-line data were correlated with the real API concentration present in the sample quantified by a previously validated off-line confocal Raman microspectroscopic method. On the other hand, in-line data were also treated in function of the concentration based on the weighing of the components in the prepared mixture. The importance of developing quantitative methods based on the use of a reference method was thus highlighted. The method was validated according to the total error approach fixing the acceptance limits at ±15% and the α risk at ±5%. This method reaches the requirements of the European Pharmacopeia norms for the uniformity of content of single-dose preparations. The validation proves that future results will be in the acceptance limits with a previously defined probability. Finally, the in-line validated method was compared with the off-line one to demonstrate its ability to be used in routine analyses. Copyright © 2017 Elsevier B.V. All rights reserved.

Analytical Method Development and Validation for the Quantification of Acetone and Isopropyl Alcohol in the Tartaric Acid Base Pellets of Dipyridamole Modified Release Capsules by Using Headspace Gas Chromatographic Technique

PubMed Central

2018-01-01

A simple, sensitive, accurate, robust headspace gas chromatographic method was developed for the quantitative determination of acetone and isopropyl alcohol in tartaric acid-based pellets of dipyridamole modified release capsules. The residual solvents acetone and isopropyl alcohol were used in the manufacturing process of the tartaric acid-based pellets of dipyridamole modified release capsules by considering the solubility of the dipyridamole and excipients in the different manufacturing stages. The method was developed and optimized by using fused silica DB-624 (30 m × 0.32 mm × 1.8 µm) column with the flame ionization detector. The method validation was carried out with regard to the guidelines for validation of analytical procedures Q2 demanded by the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH). All the validation characteristics were meeting the acceptance criteria. Hence, the developed and validated method can be applied for the intended routine analysis. PMID:29686931

Analytical Method Development and Validation for the Quantification of Acetone and Isopropyl Alcohol in the Tartaric Acid Base Pellets of Dipyridamole Modified Release Capsules by Using Headspace Gas Chromatographic Technique.

PubMed

Valavala, Sriram; Seelam, Nareshvarma; Tondepu, Subbaiah; Jagarlapudi, V Shanmukha Kumar; Sundarmurthy, Vivekanandan

2018-01-01

A simple, sensitive, accurate, robust headspace gas chromatographic method was developed for the quantitative determination of acetone and isopropyl alcohol in tartaric acid-based pellets of dipyridamole modified release capsules. The residual solvents acetone and isopropyl alcohol were used in the manufacturing process of the tartaric acid-based pellets of dipyridamole modified release capsules by considering the solubility of the dipyridamole and excipients in the different manufacturing stages. The method was developed and optimized by using fused silica DB-624 (30 m × 0.32 mm × 1.8  µ m) column with the flame ionization detector. The method validation was carried out with regard to the guidelines for validation of analytical procedures Q2 demanded by the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH). All the validation characteristics were meeting the acceptance criteria. Hence, the developed and validated method can be applied for the intended routine analysis.

ICH guidance in practice: validated reversed-phase HPLC method for the determination of active mangiferin from extracts of Mangifera indica Linn.

PubMed

Gowda, Nagaraj; Kumar, Pradeep; Panghal, Surender; Rajshree, Mashru

2010-02-01

This study presents the development and validation of a reversed-phase liquid chromatographic method for the determination of mangiferin (MGN) in alcoholic extracts of mangifera indica. A Lichrospher 100 C(18)-ODS (250 x 4.6 mm, 5 microm size) (Merck, Whitehouse Station, NJ) prepacked column and a mobile phase of potassium dihydrogen orthophosphate (0.01M) pH 2.7 +/- 0.2-acetonitrile (15:85, v/v) with the flow rate of 1 mL/min was used. MGN detection was achieved at a wavelength monitored at 254 nm with SPD-M 10A vp PDA detector or SPD 10AD vp UV detector in combination with class LC 10A software. The proposed method was validated as prescribed by International Conference on Harmonization (ICH) with respect to linearity, specificity, accuracy, precision, stability, and quantification. The method validation was realized using alcoholic extracts and raw materials of leaves and barks. All the validation parameters were within the acceptable limits, and the developed analytical method can successfully be applied for MGN determination.

A multivariate model and statistical method for validating tree grade lumber yield equations

Treesearch

Donald W. Seegrist

1975-01-01

Lumber yields within lumber grades can be described by a multivariate linear model. A method for validating lumber yield prediction equations when there are several tree grades is presented. The method is based on multivariate simultaneous test procedures.

Validation of Skills, Knowledge and Experience in Lifelong Learning in Europe

ERIC Educational Resources Information Center

Ogunleye, James

2012-01-01

The paper examines systems of validation of skills and experience as well as the main methods/tools currently used for validating skills and knowledge in lifelong learning. The paper uses mixed methods--a case study research and content analysis of European Union policy documents and frameworks--as a basis for this research. The selection of the…

Evaluating the Social Validity of the Early Start Denver Model: A Convergent Mixed Methods Study

ERIC Educational Resources Information Center

Ogilvie, Emily; McCrudden, Matthew T.

2017-01-01

An intervention has social validity to the extent that it is socially acceptable to participants and stakeholders. This pilot convergent mixed methods study evaluated parents' perceptions of the social validity of the Early Start Denver Model (ESDM), a naturalistic behavioral intervention for children with autism. It focused on whether the parents…

40 CFR Appendix D to Part 63 - Alternative Validation Procedure for EPA Waste and Wastewater Methods

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 14 2010-07-01 2010-07-01 false Alternative Validation Procedure for EPA Waste and Wastewater Methods D Appendix D to Part 63 Protection of Environment ENVIRONMENTAL... POLLUTANTS FOR SOURCE CATEGORIES (CONTINUED) Pt. 63, App. D Appendix D to Part 63—Alternative Validation...

40 CFR Appendix D to Part 63 - Alternative Validation Procedure for EPA Waste and Wastewater Methods

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 14 2011-07-01 2011-07-01 false Alternative Validation Procedure for EPA Waste and Wastewater Methods D Appendix D to Part 63 Protection of Environment ENVIRONMENTAL... POLLUTANTS FOR SOURCE CATEGORIES (CONTINUED) Pt. 63, App. D Appendix D to Part 63—Alternative Validation...

40 CFR Appendix D to Part 63 - Alternative Validation Procedure for EPA Waste and Wastewater Methods

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 15 2014-07-01 2014-07-01 false Alternative Validation Procedure for EPA Waste and Wastewater Methods D Appendix D to Part 63 Protection of Environment ENVIRONMENTAL... POLLUTANTS FOR SOURCE CATEGORIES (CONTINUED) Pt. 63, App. D Appendix D to Part 63—Alternative Validation...

40 CFR Appendix D to Part 63 - Alternative Validation Procedure for EPA Waste and Wastewater Methods

Code of Federal Regulations, 2013 CFR

2013-07-01

... 40 Protection of Environment 15 2013-07-01 2013-07-01 false Alternative Validation Procedure for EPA Waste and Wastewater Methods D Appendix D to Part 63 Protection of Environment ENVIRONMENTAL... POLLUTANTS FOR SOURCE CATEGORIES (CONTINUED) Pt. 63, App. D Appendix D to Part 63—Alternative Validation...

40 CFR Appendix D to Part 63 - Alternative Validation Procedure for EPA Waste and Wastewater Methods

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 15 2012-07-01 2012-07-01 false Alternative Validation Procedure for EPA Waste and Wastewater Methods D Appendix D to Part 63 Protection of Environment ENVIRONMENTAL... POLLUTANTS FOR SOURCE CATEGORIES (CONTINUED) Pt. 63, App. D Appendix D to Part 63—Alternative Validation...

Patterns of Cognitive Strengths and Weaknesses: Identification Rates, Agreement, and Validity for Learning Disabilities Identification

ERIC Educational Resources Information Center

Miciak, Jeremy; Fletcher, Jack M.; Stuebing, Karla K.; Vaughn, Sharon; Tolar, Tammy D.

2014-01-01

Few empirical investigations have evaluated learning disabilities (LD) identification methods based on a pattern of cognitive strengths and weaknesses (PSW). This study investigated the reliability and validity of two proposed PSW methods: the concordance/discordance method (C/DM) and cross battery assessment (XBA) method. Cognitive assessment…

Evaluation of the confusion matrix method in the validation of an automated system for measuring feeding behaviour of cattle.

PubMed

Ruuska, Salla; Hämäläinen, Wilhelmiina; Kajava, Sari; Mughal, Mikaela; Matilainen, Pekka; Mononen, Jaakko

2018-03-01

The aim of the present study was to evaluate empirically confusion matrices in device validation. We compared the confusion matrix method to linear regression and error indices in the validation of a device measuring feeding behaviour of dairy cattle. In addition, we studied how to extract additional information on classification errors with confusion probabilities. The data consisted of 12 h behaviour measurements from five dairy cows; feeding and other behaviour were detected simultaneously with a device and from video recordings. The resulting 216 000 pairs of classifications were used to construct confusion matrices and calculate performance measures. In addition, hourly durations of each behaviour were calculated and the accuracy of measurements was evaluated with linear regression and error indices. All three validation methods agreed when the behaviour was detected very accurately or inaccurately. Otherwise, in the intermediate cases, the confusion matrix method and error indices produced relatively concordant results, but the linear regression method often disagreed with them. Our study supports the use of confusion matrix analysis in validation since it is robust to any data distribution and type of relationship, it makes a stringent evaluation of validity, and it offers extra information on the type and sources of errors. Copyright © 2018 Elsevier B.V. All rights reserved.

A practical approach for linearity assessment of calibration curves under the International Union of Pure and Applied Chemistry (IUPAC) guidelines for an in-house validation of method of analysis.

PubMed

Sanagi, M Marsin; Nasir, Zalilah; Ling, Susie Lu; Hermawan, Dadan; Ibrahim, Wan Aini Wan; Naim, Ahmedy Abu

2010-01-01

Linearity assessment as required in method validation has always been subject to different interpretations and definitions by various guidelines and protocols. However, there are very limited applicable implementation procedures that can be followed by a laboratory chemist in assessing linearity. Thus, this work proposes a simple method for linearity assessment in method validation by a regression analysis that covers experimental design, estimation of the parameters, outlier treatment, and evaluation of the assumptions according to the International Union of Pure and Applied Chemistry guidelines. The suitability of this procedure was demonstrated by its application to an in-house validation for the determination of plasticizers in plastic food packaging by GC.

Verification, Validation, and Solution Quality in Computational Physics: CFD Methods Applied to Ice Sheet Physics

NASA Technical Reports Server (NTRS)

Thompson, David E.

2005-01-01

Procedures and methods for veri.cation of coding algebra and for validations of models and calculations used in the aerospace computational fluid dynamics (CFD) community would be ef.cacious if used by the glacier dynamics modeling community. This paper presents some of those methods, and how they might be applied to uncertainty management supporting code veri.cation and model validation for glacier dynamics. The similarities and differences between their use in CFD analysis and the proposed application of these methods to glacier modeling are discussed. After establishing sources of uncertainty and methods for code veri.cation, the paper looks at a representative sampling of veri.cation and validation efforts that are underway in the glacier modeling community, and establishes a context for these within an overall solution quality assessment. Finally, a vision of a new information architecture and interactive scienti.c interface is introduced and advocated.

Development and validation of a UPLC-MS/MS method for simultaneous determination of fotagliptin and its two major metabolites in human plasma and urine.

PubMed

Wang, Zhenlei; Jiang, Ji; Hu, Pei; Zhao, Qian

2017-02-01

Fotagliptin is a novel dipeptidyl peptidase IV inhibitor under clinical development for the treatment of Type II diabetes mellitus. The objective of this study was to develop and validate a specific and sensitive ultra-performance liquid chromatography (UPLC)-MS/MS method for simultaneous determination of fotagliptin and its two major metabolites in human plasma and urine. Methodology & results: After being pretreated using an automatized procedure, the plasma and urine samples were separated and detected using a UPLC-ESI-MS/MS method, which was validated following the international guidelines. A selective and sensitive UPLC-MS/MS method was first developed and validated for quantifying fotagliptin and its metabolite in human plasma and urine. The method was successfully applied to support the clinical study of fotagliptin in Chinese healthy subjects.

A brief measure of attitudes toward mixed methods research in psychology

PubMed Central

Roberts, Lynne D.; Povee, Kate

2014-01-01

The adoption of mixed methods research in psychology has trailed behind other social science disciplines. Teaching psychology students, academics, and practitioners about mixed methodologies may increase the use of mixed methods within the discipline. However, tailoring and evaluating education and training in mixed methodologies requires an understanding of, and way of measuring, attitudes toward mixed methods research in psychology. To date, no such measure exists. In this article we present the development and initial validation of a new measure: Attitudes toward Mixed Methods Research in Psychology. A pool of 42 items developed from previous qualitative research on attitudes toward mixed methods research along with validation measures was administered via an online survey to a convenience sample of 274 psychology students, academics and psychologists. Principal axis factoring with varimax rotation on a subset of the sample produced a four-factor, 12-item solution. Confirmatory factor analysis on a separate subset of the sample indicated that a higher order four factor model provided the best fit to the data. The four factors; ‘Limited Exposure,’ ‘(in)Compatibility,’ ‘Validity,’ and ‘Tokenistic Qualitative Component’; each have acceptable internal reliability. Known groups validity analyses based on preferred research orientation and self-rated mixed methods research skills, and convergent and divergent validity analyses based on measures of attitudes toward psychology as a science and scientist and practitioner orientation, provide initial validation of the measure. This brief, internally reliable measure can be used in assessing attitudes toward mixed methods research in psychology, measuring change in attitudes as part of the evaluation of mixed methods education, and in larger research programs. PMID:25429281

Using digital photography in a clinical setting: a valid, accurate, and applicable method to assess food intake.

PubMed

Winzer, Eva; Luger, Maria; Schindler, Karin

2018-06-01

Regular monitoring of food intake is hardly integrated in clinical routine. Therefore, the aim was to examine the validity, accuracy, and applicability of an appropriate and also quick and easy-to-use tool for recording food intake in a clinical setting. Two digital photography methods, the postMeal method with a picture after the meal, the pre-postMeal method with a picture before and after the meal, and the visual estimation method (plate diagram; PD) were compared against the reference method (weighed food records; WFR). A total of 420 dishes from lunch (7 weeks) were estimated with both photography methods and the visual method. Validity, applicability, accuracy, and precision of the estimation methods, and additionally food waste, macronutrient composition, and energy content were examined. Tests of validity revealed stronger correlations for photography methods (postMeal: r = 0.971, p < 0.001; pre-postMeal: r = 0.995, p < 0.001) compared to the visual estimation method (r = 0.810; p < 0.001). The pre-postMeal method showed smaller variability (bias < 1 g) and also smaller overestimation and underestimation. This method accurately and precisely estimated portion sizes in all food items. Furthermore, the total food waste was 22% for lunch over the study period. The highest food waste was observed in salads and the lowest in desserts. The pre-postMeal digital photography method is valid, accurate, and applicable in monitoring food intake in clinical setting, which enables a quantitative and qualitative dietary assessment. Thus, nutritional care might be initiated earlier. This method might be also advantageous for quantitative and qualitative evaluation of food waste, with a resultantly reduction in costs.

Vacuum decay container closure integrity leak test method development and validation for a lyophilized product-package system.

PubMed

Patel, Jayshree; Mulhall, Brian; Wolf, Heinz; Klohr, Steven; Guazzo, Dana Morton

2011-01-01

A leak test performed according to ASTM F2338-09 Standard Test Method for Nondestructive Detection of Leaks in Packages by Vacuum Decay Method was developed and validated for container-closure integrity verification of a lyophilized product in a parenteral vial package system. This nondestructive leak test method is intended for use in manufacturing as an in-process package integrity check, and for testing product stored on stability in lieu of sterility tests. Method development and optimization challenge studies incorporated artificially defective packages representing a range of glass vial wall and sealing surface defects, as well as various elastomeric stopper defects. Method validation required 3 days of random-order replicate testing of a test sample population of negative-control, no-defect packages and positive-control, with-defect packages. Positive-control packages were prepared using vials each with a single hole laser-drilled through the glass vial wall. Hole creation and hole size certification was performed by Lenox Laser. Validation study results successfully demonstrated the vacuum decay leak test method's ability to accurately and reliably detect those packages with laser-drilled holes greater than or equal to approximately 5 μm in nominal diameter. All development and validation studies were performed at Whitehouse Analytical Laboratories in Whitehouse, NJ, under the direction of consultant Dana Guazzo of RxPax, LLC, using a VeriPac 455 Micro Leak Test System by Packaging Technologies & Inspection (Tuckahoe, NY). Bristol Myers Squibb (New Brunswick, NJ) fully subsidized all work. A leak test performed according to ASTM F2338-09 Standard Test Method for Nondestructive Detection of Leaks in Packages by Vacuum Decay Method was developed and validated to detect defects in stoppered vial packages containing lyophilized product for injection. This nondestructive leak test method is intended for use in manufacturing as an in-process package integrity check, and for testing product stored on stability in lieu of sterility tests. Test method validation study results proved the method capable of detecting holes laser-drilled through the glass vial wall greater than or equal to 5 μm in nominal diameter. Total test time is less than 1 min per package. All method development and validation studies were performed at Whitehouse Analytical Laboratories in Whitehouse, NJ, under the direction of consultant Dana Guazzo of RxPax, LLC, using a VeriPac 455 Micro Leak Test System by Packaging Technologies & Inspection (Tuckahoe, NY). Bristol Myers Squibb (New Brunswick, NJ) fully subsidized all work.

Simulation verification techniques study: Simulation performance validation techniques document. [for the space shuttle system

NASA Technical Reports Server (NTRS)

Duncan, L. M.; Reddell, J. P.; Schoonmaker, P. B.

1975-01-01

Techniques and support software for the efficient performance of simulation validation are discussed. Overall validation software structure, the performance of validation at various levels of simulation integration, guidelines for check case formulation, methods for real time acquisition and formatting of data from an all up operational simulator, and methods and criteria for comparison and evaluation of simulation data are included. Vehicle subsystems modules, module integration, special test requirements, and reference data formats are also described.

Empirical Performance of Cross-Validation With Oracle Methods in a Genomics Context

PubMed Central

Martinez, Josue G.; Carroll, Raymond J.; Müller, Samuel; Sampson, Joshua N.; Chatterjee, Nilanjan

2012-01-01

When employing model selection methods with oracle properties such as the smoothly clipped absolute deviation (SCAD) and the Adaptive Lasso, it is typical to estimate the smoothing parameter by m-fold cross-validation, for example, m = 10. In problems where the true regression function is sparse and the signals large, such cross-validation typically works well. However, in regression modeling of genomic studies involving Single Nucleotide Polymorphisms (SNP), the true regression functions, while thought to be sparse, do not have large signals. We demonstrate empirically that in such problems, the number of selected variables using SCAD and the Adaptive Lasso, with 10-fold cross-validation, is a random variable that has considerable and surprising variation. Similar remarks apply to non-oracle methods such as the Lasso. Our study strongly questions the suitability of performing only a single run of m-fold cross-validation with any oracle method, and not just the SCAD and Adaptive Lasso. PMID:22347720

STATISTICAL VALIDATION OF SULFATE QUANTIFICATION METHODS USED FOR ANALYSIS OF ACID MINE DRAINAGE

EPA Science Inventory

Turbidimetric method (TM), ion chromatography (IC) and inductively coupled plasma atomic emission spectrometry (ICP-AES) with and without acid digestion have been compared and validated for the determination of sulfate in mining wastewater. Analytical methods were chosen to compa...

Validated spectrophotometric method for the determination, spectroscopic characterization and thermal structural analysis of duloxetine with 1,2-naphthoquinone-4-sulphonate

NASA Astrophysics Data System (ADS)

Ulu, Sevgi Tatar; Elmali, Fikriye Tuncel

2012-03-01

A novel, selective, sensitive and simple spectrophotometric method was developed and validated for the determination of the antidepressant duloxetine hydrochloride in pharmaceutical preparation. The method was based on the reaction of duloxetine hydrochloride with 1,2-naphthoquinone-4-sulphonate (NQS) in alkaline media to yield orange colored product. The formation of this complex was also confirmed by UV-visible, FTIR, 1H NMR, Mass spectra techniques and thermal analysis. This method was validated for various parameters according to ICH guidelines. Beer's law is obeyed in a range of 5.0-60 μg/mL at the maximum absorption wavelength of 480 nm. The detection limit is 0.99 μg/mL and the recovery rate is in a range of 98.10-99.57%. The proposed methods was validated and applied to the determination of duloxetine hydrochloride in pharmaceutical preparation. The results were statistically analyzed and compared to those of a reference UV spectrophotometric method.

[Validation and verfication of microbiology methods].

PubMed

Camaró-Sala, María Luisa; Martínez-García, Rosana; Olmos-Martínez, Piedad; Catalá-Cuenca, Vicente; Ocete-Mochón, María Dolores; Gimeno-Cardona, Concepción

2015-01-01

Clinical microbiologists should ensure, to the maximum level allowed by the scientific and technical development, the reliability of the results. This implies that, in addition to meeting the technical criteria to ensure their validity, they must be performed with a number of conditions that allows comparable results to be obtained, regardless of the laboratory that performs the test. In this sense, the use of recognized and accepted reference methodsis the most effective tool for these guarantees. The activities related to verification and validation of analytical methods has become very important, as there is continuous development, as well as updating techniques and increasingly complex analytical equipment, and an interest of professionals to ensure quality processes and results. The definitions of validation and verification are described, along with the different types of validation/verification, and the types of methods, and the level of validation necessary depending on the degree of standardization. The situations in which validation/verification is mandatory and/or recommended is discussed, including those particularly related to validation in Microbiology. It stresses the importance of promoting the use of reference strains as controls in Microbiology and the use of standard controls, as well as the importance of participation in External Quality Assessment programs to demonstrate technical competence. The emphasis is on how to calculate some of the parameters required for validation/verification, such as the accuracy and precision. The development of these concepts can be found in the microbiological process SEIMC number 48: «Validation and verification of microbiological methods» www.seimc.org/protocols/microbiology. Copyright © 2013 Elsevier España, S.L.U. y Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.

Multiple Versus Single Set Validation of Multivariate Models to Avoid Mistakes.

PubMed

Harrington, Peter de Boves

2018-01-02

Validation of multivariate models is of current importance for a wide range of chemical applications. Although important, it is neglected. The common practice is to use a single external validation set for evaluation. This approach is deficient and may mislead investigators with results that are specific to the single validation set of data. In addition, no statistics are available regarding the precision of a derived figure of merit (FOM). A statistical approach using bootstrapped Latin partitions is advocated. This validation method makes an efficient use of the data because each object is used once for validation. It was reviewed a decade earlier but primarily for the optimization of chemometric models this review presents the reasons it should be used for generalized statistical validation. Average FOMs with confidence intervals are reported and powerful, matched-sample statistics may be applied for comparing models and methods. Examples demonstrate the problems with single validation sets.

Validity and Interrater Reliability of the Visual Quarter-Waste Method for Assessing Food Waste in Middle School and High School Cafeteria Settings.

PubMed

Getts, Katherine M; Quinn, Emilee L; Johnson, Donna B; Otten, Jennifer J

2017-11-01

Measuring food waste (ie, plate waste) in school cafeterias is an important tool to evaluate the effectiveness of school nutrition policies and interventions aimed at increasing consumption of healthier meals. Visual assessment methods are frequently applied in plate waste studies because they are more convenient than weighing. The visual quarter-waste method has become a common tool in studies of school meal waste and consumption, but previous studies of its validity and reliability have used correlation coefficients, which measure association but not necessarily agreement. The aims of this study were to determine, using a statistic measuring interrater agreement, whether the visual quarter-waste method is valid and reliable for assessing food waste in a school cafeteria setting when compared with the gold standard of weighed plate waste. To evaluate validity, researchers used the visual quarter-waste method and weighed food waste from 748 trays at four middle schools and five high schools in one school district in Washington State during May 2014. To assess interrater reliability, researcher pairs independently assessed 59 of the same trays using the visual quarter-waste method. Both validity and reliability were assessed using a weighted κ coefficient. For validity, as compared with the measured weight, 45% of foods assessed using the visual quarter-waste method were in almost perfect agreement, 42% of foods were in substantial agreement, 10% were in moderate agreement, and 3% were in slight agreement. For interrater reliability between pairs of visual assessors, 46% of foods were in perfect agreement, 31% were in almost perfect agreement, 15% were in substantial agreement, and 8% were in moderate agreement. These results suggest that the visual quarter-waste method is a valid and reliable tool for measuring plate waste in school cafeteria settings. Copyright © 2017 Academy of Nutrition and Dietetics. Published by Elsevier Inc. All rights reserved.

Predictive Validity of Measures of the Pathfinder Scaling Algorithm on Programming Performance: Alternative Assessment Strategy for Programming Education

ERIC Educational Resources Information Center

Lau, Wilfred W. F.; Yuen, Allan H. K.

2009-01-01

Recent years have seen a shift in focus from assessment of learning to assessment for learning and the emergence of alternative assessment methods. However, the reliability and validity of these methods as assessment tools are still questionable. In this article, we investigated the predictive validity of measures of the Pathfinder Scaling…

Validation of an Instrument to Measure High School Students' Attitudes toward Fitness Testing

ERIC Educational Resources Information Center

Mercier, Kevin; Silverman, Stephen

2014-01-01

Purpose: The purpose of this investigation was to develop an instrument that has scores that are valid and reliable for measuring students' attitudes toward fitness testing. Method: The method involved the following steps: (a) an elicitation study, (b) item development, (c) a pilot study, and (d) a validation study. The pilot study included 427…

Sensitivity-Uncertainty Based Nuclear Criticality Safety Validation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Brown, Forrest B.

2016-09-20

These are slides from a seminar given to the University of Mexico Nuclear Engineering Department. Whisper is a statistical analysis package developed to support nuclear criticality safety validation. It uses the sensitivity profile data for an application as computed by MCNP6 along with covariance files for the nuclear data to determine a baseline upper-subcritical-limit for the application. Whisper and its associated benchmark files are developed and maintained as part of MCNP6, and will be distributed with all future releases of MCNP6. Although sensitivity-uncertainty methods for NCS validation have been under development for 20 years, continuous-energy Monte Carlo codes such asmore » MCNP could not determine the required adjoint-weighted tallies for sensitivity profiles. The recent introduction of the iterated fission probability method into MCNP led to the rapid development of sensitivity analysis capabilities for MCNP6 and the development of Whisper. Sensitivity-uncertainty based methods represent the future for NCS validation – making full use of today’s computer power to codify past approaches based largely on expert judgment. Validation results are defensible, auditable, and repeatable as needed with different assumptions and process models. The new methods can supplement, support, and extend traditional validation approaches.« less

Validation of laboratory-scale recycling test method of paper PSA label products

Treesearch

Carl Houtman; Karen Scallon; Richard Oldack

2008-01-01

Starting with test methods and a specification developed by the U.S. Postal Service (USPS) Environmentally Benign Pressure Sensitive Adhesive Postage Stamp Program, a laboratory-scale test method and a specification were developed and validated for pressure-sensitive adhesive labels, By comparing results from this new test method and pilot-scale tests, which have been...

The Method Effect in Communicative Testing.

ERIC Educational Resources Information Center

Canale, Michael

1981-01-01

A focus on test validity includes a consideration of the way a test measures that which it proposes to test; in other words, the validity of a test depends on method as well as content. This paper examines three areas of concern: (1) some features of communication that test method should reflect, (2) the main components of method, and (3) some…

Introduction to Validation of Analytical Methods: Potentiometric Determination of CO[subscript 2

ERIC Educational Resources Information Center

Hipólito-Nájera, A. Ricardo; Moya-Hernandez, M. Rosario; Gomez-Balderas, Rodolfo; Rojas-Hernandez, Alberto; Romero-Romo, Mario

2017-01-01

Validation of analytical methods is a fundamental subject for chemical analysts working in chemical industries. These methods are also relevant for pharmaceutical enterprises, biotechnology firms, analytical service laboratories, government departments, and regulatory agencies. Therefore, for undergraduate students enrolled in majors in the field…

Development and validation of reversed phase high performance liquid chromatography method for determination of dexpanthenol in pharmaceutical formulations.

PubMed

Kulikov, A U; Zinchenko, A A

2007-02-19

This paper describes the validation of an isocratic HPLC method for the assay of dexpanthenol in aerosol and gel. The method employs the Vydac Proteins C4 column with a mobile phase of aqueous solution of trifluoroacetic acid and UV detection at 206 nm. A linear response (r>0.9999) was observed in the range of 13.0-130 microg mL(-1). The method shows good recoveries and intra and inter-day relative standard deviations were less than 1.0%. Validation parameters as specificity, accuracy and robustness were also determined. The method can be used for dexpanthenol assay of panthenol aerosol and gel with dexpanthenol as the method separates dexpanthenol from aerosol or gel excipients.

A validated UHPLC-tandem mass spectrometry method for quantitative analysis of flavonolignans in milk thistle (Silybum marianum) extracts.

PubMed

Graf, Tyler N; Cech, Nadja B; Polyak, Stephen J; Oberlies, Nicholas H

2016-07-15

Validated methods are needed for the analysis of natural product secondary metabolites. These methods are particularly important to translate in vitro observations to in vivo studies. Herein, a method is reported for the analysis of the key secondary metabolites, a series of flavonolignans and a flavonoid, from an extract prepared from the seeds of milk thistle [Silybum marianum (L.) Gaertn. (Asteraceae)]. This report represents the first UHPLC MS-MS method validated for quantitative analysis of these compounds. The method takes advantage of the excellent resolution achievable with UHPLC to provide a complete analysis in less than 7min. The method is validated using both UV and MS detectors, making it applicable in laboratories with different types of analytical instrumentation available. Lower limits of quantitation achieved with this method range from 0.0400μM to 0.160μM with UV and from 0.0800μM to 0.160μM with MS. The new method is employed to evaluate variability in constituent composition in various commercial S. marianum extracts, and to show that storage of the milk thistle compounds in DMSO leads to degradation. Copyright © 2016 Elsevier B.V. All rights reserved.

Elaboration and validation of the method for the quantification of the emetic toxin of Bacillus cereus as described in EN-ISO 18465 - Microbiology of the food chain - Quantitative determination of emetic toxin (cereulide) using LC-MS/MS.

PubMed

In 't Veld, P H; van der Laak, L F J; van Zon, M; Biesta-Peters, E G

2018-04-12

A method for the quantification of the Bacillus cereus emetic toxin (cereulide) was developed and validated. The method principle is based on LC-MS as this is the most sensitive and specific method for cereulide. Therefore the study design is different from the microbiological methods validated under this mandate. As the method had to be developed a two stage validation study approach was used. The first stage (pre-study) focussed on the method applicability and the experience of the laboratories with the method. Based on the outcome of the pre-study and comments received during voting at CEN and ISO level a final method was agreed to be used for the second stage the (final) validation of the method. In the final (validation) study samples of cooked rice (both artificially contaminated with cereulide or contaminated with B. cereus for production of cereulide in the rice) and 6 other food matrices (fried rice dish, cream pastry with chocolate, hotdog sausage, mini pancakes, vanilla custard and infant formula) were used. All these samples were spiked by the participating laboratories using standard solutions of cereulide supplied by the organising laboratory. The results of the study indicate that the method is fit for purpose. Repeatability values were obtained of 0.6 μg/kg at low level spike (ca. 5 μg/kg) and 7 to 9.6 μg/kg at high level spike (ca. 75 μg/kg). Reproducibility at low spike level ranged from 0.6 to 0.9 μg/kg and from 8.7 to 14.5 μg/kg at high spike level. Recovery from the spiked samples ranged between 96.5% for mini-pancakes to 99.3% for fries rice dish. Copyright © 2018. Published by Elsevier B.V.

[Data validation methods and discussion on Chinese materia medica resource survey].

PubMed

Zhang, Yue; Ma, Wei-Feng; Zhang, Xiao-Bo; Zhu, Shou-Dong; Guo, Lan-Ping; Wang, Xing-Xing

2013-07-01

From the beginning of the fourth national survey of the Chinese materia medica resources, there were 22 provinces have conducted pilots. The survey teams have reported immense data, it put forward the very high request to the database system construction. In order to ensure the quality, it is necessary to check and validate the data in database system. Data validation is important methods to ensure the validity, integrity and accuracy of census data. This paper comprehensively introduce the data validation system of the fourth national survey of the Chinese materia medica resources database system, and further improve the design idea and programs of data validation. The purpose of this study is to promote the survey work smoothly.

Validation of spatial variability in downscaling results from the VALUE perfect predictor experiment

NASA Astrophysics Data System (ADS)

Widmann, Martin; Bedia, Joaquin; Gutiérrez, Jose Manuel; Maraun, Douglas; Huth, Radan; Fischer, Andreas; Keller, Denise; Hertig, Elke; Vrac, Mathieu; Wibig, Joanna; Pagé, Christian; Cardoso, Rita M.; Soares, Pedro MM; Bosshard, Thomas; Casado, Maria Jesus; Ramos, Petra

2016-04-01

VALUE is an open European network to validate and compare downscaling methods for climate change research. Within VALUE a systematic validation framework to enable the assessment and comparison of both dynamical and statistical downscaling methods has been developed. In the first validation experiment the downscaling methods are validated in a setup with perfect predictors taken from the ERA-interim reanalysis for the period 1997 - 2008. This allows to investigate the isolated skill of downscaling methods without further error contributions from the large-scale predictors. One aspect of the validation is the representation of spatial variability. As part of the VALUE validation we have compared various properties of the spatial variability of downscaled daily temperature and precipitation with the corresponding properties in observations. We have used two test validation datasets, one European-wide set of 86 stations, and one higher-density network of 50 stations in Germany. Here we present results based on three approaches, namely the analysis of i.) correlation matrices, ii.) pairwise joint threshold exceedances, and iii.) regions of similar variability. We summarise the information contained in correlation matrices by calculating the dependence of the correlations on distance and deriving decorrelation lengths, as well as by determining the independent degrees of freedom. Probabilities for joint threshold exceedances and (where appropriate) non-exceedances are calculated for various user-relevant thresholds related for instance to extreme precipitation or frost and heat days. The dependence of these probabilities on distance is again characterised by calculating typical length scales that separate dependent from independent exceedances. Regionalisation is based on rotated Principal Component Analysis. The results indicate which downscaling methods are preferable if the dependency of variability at different locations is relevant for the user.

Validation of microbiological testing in cardiovascular tissue banks: results of a quality round trial.

PubMed

de By, Theo M M H; McDonald, Carl; Süßner, Susanne; Davies, Jill; Heng, Wee Ling; Jashari, Ramadan; Bogers, Ad J J C; Petit, Pieter

2017-11-01

Surgeons needing human cardiovascular tissue for implantation in their patients are confronted with cardiovascular tissue banks that use different methods to identify and decontaminate micro-organisms. To elucidate these differences, we compared the quality of processing methods in 20 tissue banks and 1 reference laboratory. We did this to validate the results for accepting or rejecting tissue. We included the decontamination methods used and the influence of antibiotic cocktails and residues with results and controls. The minor details of the processes were not included. To compare the outcomes of microbiological testing and decontamination methods of heart valve allografts in cardiovascular tissue banks, an international quality round was organized. Twenty cardiovascular tissue banks participated in this quality round. The quality round method was validated first and consisted of sending purposely contaminated human heart valve tissue samples with known micro-organisms to the participants. The participants identified the micro-organisms using their local decontamination methods. Seventeen of the 20 participants correctly identified the micro-organisms; if these samples were heart valves to be released for implantation, 3 of the 20 participants would have decided to accept their result for release. Decontamination was shown not to be effective in 13 tissue banks because of growth of the organisms after decontamination. Articles in the literature revealed that antibiotics are effective at 36°C and not, or less so, at 2-8°C. The decontamination procedure, if it is validated, will ensure that the tissue contains no known micro-organisms. This study demonstrates that the quality round method of sending contaminated tissues and assessing the results of the microbiological cultures is an effective way of validating the processes of tissue banks. Only when harmonization, based on validated methods, has been achieved, will surgeons be able to fully rely on the methods used and have confidence in the consistent sterility of the tissue grafts. Tissue banks should validate their methods so that all stakeholders can trust the outcomes. © The Author 2017. Published by Oxford University Press on behalf of the European Association for Cardio-Thoracic Surgery. All rights reserved.

A new validated method for the simultaneous determination of benzocaine, propylparaben and benzyl alcohol in a bioadhesive gel by HPLC.

PubMed

Pérez-Lozano, P; García-Montoya, E; Orriols, A; Miñarro, M; Ticó, J R; Suñé-Negre, J M

2005-10-04

A new HPLC-RP method has been developed and validated for the simultaneous determination of benzocaine, two preservatives (propylparaben (nipasol) and benzyl alcohol) and degradation products of benzocaine in a semisolid pharmaceutical dosage form (benzocaine gel). The method uses a Nucleosil 120 C18 column and gradient elution. The mobile phase consisted of a mixture of methanol and glacial acetic acid (10%, v/v) at different proportion according to a time-schedule programme, pumped at a flow rate of 2.0 ml min(-1). The DAD detector was set at 258 nm. The validation study was carried out fulfilling the ICH guidelines in order to prove that the new analytical method, meets the reliability characteristics, and these characteristics showed the capacity of analytical method to keep, throughout the time, the fundamental criteria for validation: selectivity, linearity, precision, accuracy and sensitivity. The method was applied during the quality control of benzocaine gel in order to quantify the drug (benzocaine), preservatives and degraded products and proved to be suitable for rapid and reliable quality control method.

Alternative methods to determine headwater benefits

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bao, Y.S.; Perlack, R.D.; Sale, M.J.

1997-11-10

In 1992, the Federal Energy Regulatory Commission (FERC) began using a Flow Duration Analysis (FDA) methodology to assess headwater benefits in river basins where use of the Headwater Benefits Energy Gains (HWBEG) model may not result in significant improvements in modeling accuracy. The purpose of this study is to validate the accuracy and appropriateness of the FDA method for determining energy gains in less complex basins. This report presents the results of Oak Ridge National Laboratory`s (ORNL`s) validation of the FDA method. The validation is based on a comparison of energy gains using the FDA method with energy gains calculatedmore » using the MWBEG model. Comparisons of energy gains are made on a daily and monthly basis for a complex river basin (the Alabama River Basin) and a basin that is considered relatively simple hydrologically (the Stanislaus River Basin). In addition to validating the FDA method, ORNL was asked to suggest refinements and improvements to the FDA method. Refinements and improvements to the FDA method were carried out using the James River Basin as a test case.« less

Bioanalytical method development and validation for the determination of glycine in human cerebrospinal fluid by ion-pair reversed-phase liquid chromatography-tandem mass spectrometry.

PubMed

Jiang, Jian; James, Christopher A; Wong, Philip

2016-09-05

A LC-MS/MS method has been developed and validated for the determination of glycine in human cerebrospinal fluid (CSF). The validated method used artificial cerebrospinal fluid as a surrogate matrix for calibration standards. The calibration curve range for the assay was 100-10,000ng/mL and (13)C2, (15)N-glycine was used as an internal standard (IS). Pre-validation experiments were performed to demonstrate parallelism with surrogate matrix and standard addition methods. The mean endogenous glycine concentration in a pooled human CSF determined on three days by using artificial CSF as a surrogate matrix and the method of standard addition was found to be 748±30.6 and 768±18.1ng/mL, respectively. A percentage difference of -2.6% indicated that artificial CSF could be used as a surrogate calibration matrix for the determination of glycine in human CSF. Quality control (QC) samples, except the lower limit of quantitation (LLOQ) QC and low QC samples, were prepared by spiking glycine into aliquots of pooled human CSF sample. The low QC sample was prepared from a separate pooled human CSF sample containing low endogenous glycine concentrations, while the LLOQ QC sample was prepared in artificial CSF. Standard addition was used extensively to evaluate matrix effects during validation. The validated method was used to determine the endogenous glycine concentrations in human CSF samples. Incurred sample reanalysis demonstrated reproducibility of the method. Copyright © 2016 Elsevier B.V. All rights reserved.

Low-cost extrapolation method for maximal LTE radio base station exposure estimation: test and validation.

PubMed

Verloock, Leen; Joseph, Wout; Gati, Azeddine; Varsier, Nadège; Flach, Björn; Wiart, Joe; Martens, Luc

2013-06-01

An experimental validation of a low-cost method for extrapolation and estimation of the maximal electromagnetic-field exposure from long-term evolution (LTE) radio base station installations are presented. No knowledge on downlink band occupation or service characteristics is required for the low-cost method. The method is applicable in situ. It only requires a basic spectrum analyser with appropriate field probes without the need of expensive dedicated LTE decoders. The method is validated both in laboratory and in situ, for a single-input single-output antenna LTE system and a 2×2 multiple-input multiple-output system, with low deviations in comparison with signals measured using dedicated LTE decoders.

Expression and characterization of an iron-regulated hemin-binding protein, HbpA, from Leptospira interrogans serovar Lai.

PubMed

Asuthkar, Swapna; Velineni, Sridhar; Stadlmann, Johannes; Altmann, Friedrich; Sritharan, Manjula

2007-09-01

In an earlier study, based on the ferric enterobactin receptor FepA of Escherichia coli, we identified and modeled a TonB-dependent outer membrane receptor protein (LB191) from the genome of Leptospira interrogans serovar Lai. Based on in silico analysis, we hypothesized that this protein was an iron-dependent hemin-binding protein. In this study, we provide experimental evidence to prove that this protein, termed HbpA (hemin-binding protein A), is indeed an iron-regulated hemin-binding protein. We cloned and expressed the full-length 81-kDa recombinant rHbpA protein and a truncated 55-kDa protein from L. interrogans serovar Lai, both of which bind hemin-agarose. Assay of hemin-associated peroxidase activity and spectrofluorimetric analysis provided confirmatory evidence of hemin binding by HbpA. Immunofluorescence studies by confocal microscopy and the microscopic agglutination test demonstrated the surface localization and the iron-regulated expression of HbpA in L. interrogans. Southern blot analysis confirmed our earlier observation that the hbpA gene was present only in some of the pathogenic serovars and was absent in Leptospira biflexa. Hemin-agarose affinity studies showed another hemin-binding protein with a molecular mass of approximately 44 kDa, whose expression was independent of iron levels. This protein was seen in several serovars, including nonpathogenic L. biflexa. Sequence analysis and immunoreactivity with specific antibodies showed this protein to be LipL41.

Calcium Stearate as an Effective Alternative to Poly(vinyl alcohol) in Poly-Lactic-co-Glycolic Acid Nanoparticles Synthesis.

PubMed

Cella, Claudia; Gerges, Irini; Milani, Paolo; Lenardi, Cristina; Argentiere, Simona

2017-02-13

Poly(d,l-lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) are among the most studied systems for drug and gene targeting. So far, the synthesis of stable and uniform PLGA NPs has involved the use of a large excess of polyvinyl surfactants such as poly(vinyl alcohol) (PVA) and polyvinylpyrrolidone (PVP), whose removal requires multistep purification procedures of high ecological and economic impact. Hence the development of environment-friendly and cost-effective synthetic procedures for the synthesis of PLGA NPs would effectively boost their use in clinics. This work aims to address this issue by investigating more efficacious alternatives to the so far employed polyvinyl surfactants. More specifically, we developed an innovative synthetic process to achieve stable and uniformly distributed PLGA NPs that involves the use of calcium stearate (CSt), gaining benefits of its high biocompatibility and efficacy at low concentrations and avoiding consequently expensive purification steps. With the help of minimum quantities of polysorbate 60 and sorbitane monostearate, CSt-stabilized PLGA NPs with different sizes and structures were synthesized. The influence of CSt on the encapsulation efficiency of bioactive molecules has been also investigated. The effective encapsulation of both hydrophobic (curcumin) and hydrophilic (fibrinogen labeled with Alexa647) biomolecules into NPs was demonstrated by confocal microscopy, and their release quantified by spectrofluorimetric analyses. Finally, degradation and cytotoxicity studies showed that CSt stabilized NPs were stable under physiological conditions and with good biocompatibility, thus looking promising for further investigation as controlled release devices.

Validation of biological activity testing procedure of recombinant human interleukin-7.

PubMed

Lutsenko, T N; Kovalenko, M V; Galkin, O Yu

2017-01-01

Validation procedure for method of monitoring the biological activity of reсombinant human interleukin-7 has been developed and conducted according to the requirements of national and international recommendations. This method is based on the ability of recombinant human interleukin-7 to induce proliferation of T lymphocytes. It has been shown that to control the biological activity of recombinant human interleukin-7 peripheral blood mononuclear cells (PBMCs) derived from blood or cell lines can be used. Validation charac­teristics that should be determined depend on the method, type of product or object test/measurement and biological test systems used in research. The validation procedure for the method of control of biological activity of recombinant human interleukin-7 in peripheral blood mononuclear cells showed satisfactory results on all parameters tested such as specificity, accuracy, precision and linearity.

Global Land Product Validation Protocols: An Initiative of the CEOS Working Group on Calibration and Validation to Evaluate Satellite-derived Essential Climate Variables

NASA Astrophysics Data System (ADS)

Guillevic, P. C.; Nickeson, J. E.; Roman, M. O.; camacho De Coca, F.; Wang, Z.; Schaepman-Strub, G.

2016-12-01

The Global Climate Observing System (GCOS) has specified the need to systematically produce and validate Essential Climate Variables (ECVs). The Committee on Earth Observation Satellites (CEOS) Working Group on Calibration and Validation (WGCV) and in particular its subgroup on Land Product Validation (LPV) is playing a key coordination role leveraging the international expertise required to address actions related to the validation of global land ECVs. The primary objective of the LPV subgroup is to set standards for validation methods and reporting in order to provide traceable and reliable uncertainty estimates for scientists and stakeholders. The Subgroup is comprised of 9 focus areas that encompass 10 land surface variables. The activities of each focus area are coordinated by two international co-leads and currently include leaf area index (LAI) and fraction of absorbed photosynthetically active radiation (FAPAR), vegetation phenology, surface albedo, fire disturbance, snow cover, land cover and land use change, soil moisture, land surface temperature (LST) and emissivity. Recent additions to the focus areas include vegetation indices and biomass. The development of best practice validation protocols is a core activity of CEOS LPV with the objective to standardize the evaluation of land surface products. LPV has identified four validation levels corresponding to increasing spatial and temporal representativeness of reference samples used to perform validation. Best practice validation protocols (1) provide the definition of variables, ancillary information and uncertainty metrics, (2) describe available data sources and methods to establish reference validation datasets with SI traceability, and (3) describe evaluation methods and reporting. An overview on validation best practice components will be presented based on the LAI and LST protocol efforts to date.

ASTM Validates Air Pollution Test Methods

ERIC Educational Resources Information Center

Chemical and Engineering News, 1973

1973-01-01

The American Society for Testing and Materials (ASTM) has validated six basic methods for measuring pollutants in ambient air as the first part of its Project Threshold. Aim of the project is to establish nationwide consistency in measuring pollutants; determining precision, accuracy and reproducibility of 35 standard measuring methods. (BL)

Evaluation of the repeatability and reproducibility of a suite of qPCR based microbial source tracking methods

EPA Science Inventory

Many PCR-based methods for microbial source tracking (MST) have been developed and validated within individual research laboratories. Inter-laboratory validation of these methods, however, has been minimal, and the effects of protocol standardization regimes have not been thor...

75 FR 25867 - National Toxicology Program (NTP) Interagency Center for the Evaluation of Alternative...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-05-10

... validation studies. NICEATM and ICCVAM work collaboratively to evaluate new and improved test methods... for nomination of test methods for validation studies, and guidelines for submission of test methods... for human and veterinary vaccine post-licensing potency and safety testing. Plenary and breakout...

A hydrostatic weighing method using total lung capacity and a small tank.

PubMed Central

Warner, J G; Yeater, R; Sherwood, L; Weber, K

1986-01-01

The purpose of this study was to establish the validity and reliability of a hydrostatic weighing method using total lung capacity (measuring vital capacity with a respirometer at the time of weighing) the prone position, and a small oblong tank. The validity of the method was established by comparing the TLC prone (tank) method against three hydrostatic weighing methods administered in a pool. The three methods included residual volume seated, TLC seated and TLC prone. Eighty male and female subjects were underwater weighed using each of the four methods. Validity coefficients for per cent body fat between the TLC prone (tank) method and the RV seated (pool), TLC seated (pool) and TLC prone (pool) methods were .98, .99 and .99, respectively. A randomised complete block ANOVA found significant differences between the RV seated (pool) method and each of the three TLC methods with respect to both body density and per cent body fat. The differences were negligible with respect to HW error. Reliability of the TLC prone (tank) method was established by weighing twenty subjects three different times with ten-minute time intervals between testing. Multiple correlations yielded reliability coefficients for body density and per cent body fat values of .99 and .99, respectively. It was concluded that the TLC prone (tank) method is valid, reliable and a favourable method of hydrostatic weighing. PMID:3697596

A hydrostatic weighing method using total lung capacity and a small tank.

PubMed

Warner, J G; Yeater, R; Sherwood, L; Weber, K

1986-03-01

The purpose of this study was to establish the validity and reliability of a hydrostatic weighing method using total lung capacity (measuring vital capacity with a respirometer at the time of weighing) the prone position, and a small oblong tank. The validity of the method was established by comparing the TLC prone (tank) method against three hydrostatic weighing methods administered in a pool. The three methods included residual volume seated, TLC seated and TLC prone. Eighty male and female subjects were underwater weighed using each of the four methods. Validity coefficients for per cent body fat between the TLC prone (tank) method and the RV seated (pool), TLC seated (pool) and TLC prone (pool) methods were .98, .99 and .99, respectively. A randomised complete block ANOVA found significant differences between the RV seated (pool) method and each of the three TLC methods with respect to both body density and per cent body fat. The differences were negligible with respect to HW error. Reliability of the TLC prone (tank) method was established by weighing twenty subjects three different times with ten-minute time intervals between testing. Multiple correlations yielded reliability coefficients for body density and per cent body fat values of .99 and .99, respectively. It was concluded that the TLC prone (tank) method is valid, reliable and a favourable method of hydrostatic weighing.

History and development of the Schmidt-Hunter meta-analysis methods.

PubMed

Schmidt, Frank L

2015-09-01

In this article, I provide answers to the questions posed by Will Shadish about the history and development of the Schmidt-Hunter methods of meta-analysis. In the 1970s, I headed a research program on personnel selection at the US Office of Personnel Management (OPM). After our research showed that validity studies have low statistical power, OPM felt a need for a better way to demonstrate test validity, especially in light of court cases challenging selection methods. In response, we created our method of meta-analysis (initially called validity generalization). Results showed that most of the variability of validity estimates from study to study was because of sampling error and other research artifacts such as variations in range restriction and measurement error. Corrections for these artifacts in our research and in replications by others showed that the predictive validity of most tests was high and generalizable. This conclusion challenged long-standing beliefs and so provoked resistance, which over time was overcome. The 1982 book that we published extending these methods to research areas beyond personnel selection was positively received and was followed by expanded books in 1990, 2004, and 2014. Today, these methods are being applied in a wide variety of areas. Copyright © 2015 John Wiley & Sons, Ltd.

Evaluating the statistical performance of less applied algorithms in classification of worldview-3 imagery data in an urbanized landscape

NASA Astrophysics Data System (ADS)

Ranaie, Mehrdad; Soffianian, Alireza; Pourmanafi, Saeid; Mirghaffari, Noorollah; Tarkesh, Mostafa

2018-03-01

In recent decade, analyzing the remotely sensed imagery is considered as one of the most common and widely used procedures in the environmental studies. In this case, supervised image classification techniques play a central role. Hence, taking a high resolution Worldview-3 over a mixed urbanized landscape in Iran, three less applied image classification methods including Bagged CART, Stochastic gradient boosting model and Neural network with feature extraction were tested and compared with two prevalent methods: random forest and support vector machine with linear kernel. To do so, each method was run ten time and three validation techniques was used to estimate the accuracy statistics consist of cross validation, independent validation and validation with total of train data. Moreover, using ANOVA and Tukey test, statistical difference significance between the classification methods was significantly surveyed. In general, the results showed that random forest with marginal difference compared to Bagged CART and stochastic gradient boosting model is the best performing method whilst based on independent validation there was no significant difference between the performances of classification methods. It should be finally noted that neural network with feature extraction and linear support vector machine had better processing speed than other.

Development and validation of a discriminative dissolution method for atorvastatin calcium tablets using in vivo data by LC and UV methods.

PubMed

Machado, J C; Lange, A D; Todeschini, V; Volpato, N M

2014-02-01

A dissolution method to analyze atorvastatin tablets using in vivo data for RP and test pilot (PB) was developed and validated. The appropriate conditions were determined after solubility tests using different media, and sink conditions were established. The conditions used were equipment paddle at 50 rpm and 900 mL of potassium phosphate buffer pH 6.0 as dissolution medium. In vivo release profiles were obtained from the bioequivalence study of RP and the generic candidate PB. The fraction of dose absorbed was calculated using the Loo-Riegelman method. It was necessary to use a scale factor of time similar to 6.0, to associate the values of absorbed fraction and dissolved fraction, obtaining an in vivo-in vitro correlation level A. The dissolution method to quantify the amount of drug dissolved was validated using high-performance liquid chromatography and ultraviolet spectrophotometry, and validated according to the USP protocol. The discriminative power of dissolution conditions was assessed using two different pilot batches of atorvastatin tablets (PA and PB) and RP. The dissolution test was validated and may be used as a discriminating method in quality control and in the development of the new formulations.

Analysis of Phosphonic Acids: Validation of Semi-Volatile Analysis by HPLC-MS/MS by EPA Method MS999

DOE Office of Scientific and Technical Information (OSTI.GOV)

Owens, J; Vu, A; Koester, C

The Environmental Protection Agency's (EPA) Region 5 Chicago Regional Laboratory (CRL) developed a method titled Analysis of Diisopropyl Methylphosphonate, Ethyl Hydrogen Dimethylamidophosphate, Isopropyl Methylphosphonic Acid, Methylphosphonic Acid, and Pinacolyl Methylphosphonic Acid in Water by Multiple Reaction Monitoring Liquid Chromatography/Tandem Mass Spectrometry: EPA Version MS999. This draft standard operating procedure (SOP) was distributed to multiple EPA laboratories and to Lawrence Livermore National Laboratory, which was tasked to serve as a reference laboratory for EPA's Environmental Reference Laboratory Network (ERLN) and to develop and validate analytical procedures. The primary objective of this study was to validate and verify the analytical procedures describedmore » in EPA Method MS999 for analysis of the listed phosphonic acids and surrogates in aqueous samples. The gathered data from this validation study will be used to: (1) demonstrate analytical method performance; (2) generate quality control acceptance criteria; and (3) revise the SOP to provide a validated method that would be available for use during a homeland security event. The data contained in this report will be compiled, by EPA CRL, with data generated by other EPA Regional laboratories so that performance metrics of EPA Method MS999 can be determined.« less

Testing and Validation of Computational Methods for Mass Spectrometry.

PubMed

Gatto, Laurent; Hansen, Kasper D; Hoopmann, Michael R; Hermjakob, Henning; Kohlbacher, Oliver; Beyer, Andreas

2016-03-04

High-throughput methods based on mass spectrometry (proteomics, metabolomics, lipidomics, etc.) produce a wealth of data that cannot be analyzed without computational methods. The impact of the choice of method on the overall result of a biological study is often underappreciated, but different methods can result in very different biological findings. It is thus essential to evaluate and compare the correctness and relative performance of computational methods. The volume of the data as well as the complexity of the algorithms render unbiased comparisons challenging. This paper discusses some problems and challenges in testing and validation of computational methods. We discuss the different types of data (simulated and experimental validation data) as well as different metrics to compare methods. We also introduce a new public repository for mass spectrometric reference data sets ( http://compms.org/RefData ) that contains a collection of publicly available data sets for performance evaluation for a wide range of different methods.

Validity and reliability of a method for assessment of cervical vertebral maturation.

PubMed

Zhao, Xiao-Guang; Lin, Jiuxiang; Jiang, Jiu-Hui; Wang, Qingzhu; Ng, Sut Hong

2012-03-01

To evaluate the validity and reliability of the cervical vertebral maturation (CVM) method with a longitudinal sample. Eighty-six cephalograms from 18 subjects (5 males and 13 females) were selected from the longitudinal database. Total mandibular length was measured on each film; an increased rate served as the gold standard in examination of the validity of the CVM method. Eleven orthodontists, after receiving intensive training in the CVM method, evaluated all films twice. Kendall's W and the weighted kappa statistic were employed. Kendall's W values were higher than 0.8 at both times, indicating strong interobserver reproducibility, but interobserver agreement was documented twice at less than 50%. A wide range of intraobserver agreement was noted (40.7%-79.1%), and substantial intraobserver reproducibility was proved by kappa values (0.53-0.86). With regard to validity, moderate agreement was reported between the gold standard and observer staging at the initial time (kappa values 0.44-0.61). However, agreement seemed to be unacceptable for clinical use, especially in cervical stage 3 (26.8%). Even though the validity and reliability of the CVM method proved statistically acceptable, we suggest that many other growth indicators should be taken into consideration in evaluating adolescent skeletal maturation.

Working towards accreditation by the International Standards Organization 15189 Standard: how to validate an in-house developed method an example of lead determination in whole blood by electrothermal atomic absorption spectrometry.

PubMed

Garcia Hejl, Carine; Ramirez, Jose Manuel; Vest, Philippe; Chianea, Denis; Renard, Christophe

2014-09-01

Laboratories working towards accreditation by the International Standards Organization (ISO) 15189 standard are required to demonstrate the validity of their analytical methods. The different guidelines set by various accreditation organizations make it difficult to provide objective evidence that an in-house method is fit for the intended purpose. Besides, the required performance characteristics tests and acceptance criteria are not always detailed. The laboratory must choose the most suitable validation protocol and set the acceptance criteria. Therefore, we propose a validation protocol to evaluate the performance of an in-house method. As an example, we validated the process for the detection and quantification of lead in whole blood by electrothermal absorption spectrometry. The fundamental parameters tested were, selectivity, calibration model, precision, accuracy (and uncertainty of measurement), contamination, stability of the sample, reference interval, and analytical interference. We have developed a protocol that has been applied successfully to quantify lead in whole blood by electrothermal atomic absorption spectrometry (ETAAS). In particular, our method is selective, linear, accurate, and precise, making it suitable for use in routine diagnostics.

Semi-automating the manual literature search for systematic reviews increases efficiency.

PubMed

Chapman, Andrea L; Morgan, Laura C; Gartlehner, Gerald

2010-03-01

To minimise retrieval bias, manual literature searches are a key part of the search process of any systematic review. Considering the need to have accurate information, valid results of the manual literature search are essential to ensure scientific standards; likewise efficient approaches that minimise the amount of personnel time required to conduct a manual literature search are of great interest. The objective of this project was to determine the validity and efficiency of a new manual search method that utilises the scopus database. We used the traditional manual search approach as the gold standard to determine the validity and efficiency of the proposed scopus method. Outcome measures included completeness of article detection and personnel time involved. Using both methods independently, we compared the results based on accuracy of the results, validity and time spent conducting the search, efficiency. Regarding accuracy, the scopus method identified the same studies as the traditional approach indicating its validity. In terms of efficiency, using scopus led to a time saving of 62.5% compared with the traditional approach (3 h versus 8 h). The scopus method can significantly improve the efficiency of manual searches and thus of systematic reviews.

Comprehensive validation scheme for in situ fiber optics dissolution method for pharmaceutical drug product testing.

PubMed

Mirza, Tahseen; Liu, Qian Julie; Vivilecchia, Richard; Joshi, Yatindra

2009-03-01

There has been a growing interest during the past decade in the use of fiber optics dissolution testing. Use of this novel technology is mainly confined to research and development laboratories. It has not yet emerged as a tool for end product release testing despite its ability to generate in situ results and efficiency improvement. One potential reason may be the lack of clear validation guidelines that can be applied for the assessment of suitability of fiber optics. This article describes a comprehensive validation scheme and development of a reliable, robust, reproducible and cost-effective dissolution test using fiber optics technology. The test was successfully applied for characterizing the dissolution behavior of a 40-mg immediate-release tablet dosage form that is under development at Novartis Pharmaceuticals, East Hanover, New Jersey. The method was validated for the following parameters: linearity, precision, accuracy, specificity, and robustness. In particular, robustness was evaluated in terms of probe sampling depth and probe orientation. The in situ fiber optic method was found to be comparable to the existing manual sampling dissolution method. Finally, the fiber optic dissolution test was successfully performed by different operators on different days, to further enhance the validity of the method. The results demonstrate that the fiber optics technology can be successfully validated for end product dissolution/release testing. (c) 2008 Wiley-Liss, Inc. and the American Pharmacists Association

Proposal of a method for the evaluation of inaccuracy of home sphygmomanometers.

PubMed

Akpolat, Tekin

2009-10-01

There is no formal protocol for evaluating the individual accuracy of home sphygmomanometers. The aims of this study were to propose a method for achieving accuracy in automated home sphygmomanometers and to test the applicability of the defined method. The purposes of this method were to avoid major inaccuracies and to estimate the optimal circumstance for individual accuracy. The method has three stages and sequential measurement of blood pressure is used. The tested devices were categorized into four groups: accurate, acceptable, inaccurate and very inaccurate (major inaccuracy). The defined method takes approximately 10 min (excluding relaxation time) and was tested on three different occasions. The application of the method has shown that inaccuracy is a common problem among non-tested devices, that validated devices are superior to those that are non-validated or whose validation status is unknown, that major inaccuracy is common, especially in non-tested devices and that validation does not guarantee individual accuracy. A protocol addressing the accuracy of a particular sphygmomanometer in an individual patient is required, and a practical method has been suggested to achieve this. This method can be modified, but the main idea and approach should be preserved unless a better method is proposed. The purchase of validated devices and evaluation of accuracy for the purchased device in an individual patient will improve the monitoring of self-measurement of blood pressure at home. This study addresses device inaccuracy, but errors related to the patient, observer or blood pressure measurement technique should not be underestimated, and strict adherence to the manufacturer's instructions is essential.

Correcting for Optimistic Prediction in Small Data Sets

PubMed Central

Smith, Gordon C. S.; Seaman, Shaun R.; Wood, Angela M.; Royston, Patrick; White, Ian R.

2014-01-01

The C statistic is a commonly reported measure of screening test performance. Optimistic estimation of the C statistic is a frequent problem because of overfitting of statistical models in small data sets, and methods exist to correct for this issue. However, many studies do not use such methods, and those that do correct for optimism use diverse methods, some of which are known to be biased. We used clinical data sets (United Kingdom Down syndrome screening data from Glasgow (1991–2003), Edinburgh (1999–2003), and Cambridge (1990–2006), as well as Scottish national pregnancy discharge data (2004–2007)) to evaluate different approaches to adjustment for optimism. We found that sample splitting, cross-validation without replication, and leave-1-out cross-validation produced optimism-adjusted estimates of the C statistic that were biased and/or associated with greater absolute error than other available methods. Cross-validation with replication, bootstrapping, and a new method (leave-pair-out cross-validation) all generated unbiased optimism-adjusted estimates of the C statistic and had similar absolute errors in the clinical data set. Larger simulation studies confirmed that all 3 methods performed similarly with 10 or more events per variable, or when the C statistic was 0.9 or greater. However, with lower events per variable or lower C statistics, bootstrapping tended to be optimistic but with lower absolute and mean squared errors than both methods of cross-validation. PMID:24966219

Towards a full integration of optimization and validation phases: An analytical-quality-by-design approach.

PubMed

Hubert, C; Houari, S; Rozet, E; Lebrun, P; Hubert, Ph

2015-05-22

When using an analytical method, defining an analytical target profile (ATP) focused on quantitative performance represents a key input, and this will drive the method development process. In this context, two case studies were selected in order to demonstrate the potential of a quality-by-design (QbD) strategy when applied to two specific phases of the method lifecycle: the pre-validation study and the validation step. The first case study focused on the improvement of a liquid chromatography (LC) coupled to mass spectrometry (MS) stability-indicating method by the means of the QbD concept. The design of experiments (DoE) conducted during the optimization step (i.e. determination of the qualitative design space (DS)) was performed a posteriori. Additional experiments were performed in order to simultaneously conduct the pre-validation study to assist in defining the DoE to be conducted during the formal validation step. This predicted protocol was compared to the one used during the formal validation. A second case study based on the LC/MS-MS determination of glucosamine and galactosamine in human plasma was considered in order to illustrate an innovative strategy allowing the QbD methodology to be incorporated during the validation phase. An operational space, defined by the qualitative DS, was considered during the validation process rather than a specific set of working conditions as conventionally performed. Results of all the validation parameters conventionally studied were compared to those obtained with this innovative approach for glucosamine and galactosamine. Using this strategy, qualitative and quantitative information were obtained. Consequently, an analyst using this approach would be able to select with great confidence several working conditions within the operational space rather than a given condition for the routine use of the method. This innovative strategy combines both a learning process and a thorough assessment of the risk involved. Copyright © 2015 Elsevier B.V. All rights reserved.

Real-Time PCR Method for Detection of Salmonella spp. in Environmental Samples.

PubMed

Kasturi, Kuppuswamy N; Drgon, Tomas

2017-07-15

The methods currently used for detecting Salmonella in environmental samples require 2 days to produce results and have limited sensitivity. Here, we describe the development and validation of a real-time PCR Salmonella screening method that produces results in 18 to 24 h. Primers and probes specific to the gene invA , group D, and Salmonella enterica serovar Enteritidis organisms were designed and evaluated for inclusivity and exclusivity using a panel of 329 Salmonella isolates representing 126 serovars and 22 non- Salmonella organisms. The invA - and group D-specific sets identified all the isolates accurately. The PCR method had 100% inclusivity and detected 1 to 2 copies of Salmonella DNA per reaction. Primers specific for Salmonella -differentiating fragment 1 (Sdf-1) in conjunction with the group D set had 100% inclusivity for 32 S Enteritidis isolates and 100% exclusivity for the 297 non-Enteritidis Salmonella isolates. Single-laboratory validation performed on 1,741 environmental samples demonstrated that the PCR method detected 55% more positives than the V itek i mmuno d iagnostic a ssay s ystem (VIDAS) method. The PCR results correlated well with the culture results, and the method did not report any false-negative results. The receiver operating characteristic (ROC) analysis documented excellent agreement between the results from the culture and PCR methods (area under the curve, 0.90; 95% confidence interval of 0.76 to 1.0) confirming the validity of the PCR method. IMPORTANCE This validated PCR method detects 55% more positives for Salmonella in half the time required for the reference method, VIDAS. The validated PCR method will help to strengthen public health efforts through rapid screening of Salmonella spp. in environmental samples.

Real-Time PCR Method for Detection of Salmonella spp. in Environmental Samples

PubMed Central

Drgon, Tomas

2017-01-01

ABSTRACT The methods currently used for detecting Salmonella in environmental samples require 2 days to produce results and have limited sensitivity. Here, we describe the development and validation of a real-time PCR Salmonella screening method that produces results in 18 to 24 h. Primers and probes specific to the gene invA, group D, and Salmonella enterica serovar Enteritidis organisms were designed and evaluated for inclusivity and exclusivity using a panel of 329 Salmonella isolates representing 126 serovars and 22 non-Salmonella organisms. The invA- and group D-specific sets identified all the isolates accurately. The PCR method had 100% inclusivity and detected 1 to 2 copies of Salmonella DNA per reaction. Primers specific for Salmonella-differentiating fragment 1 (Sdf-1) in conjunction with the group D set had 100% inclusivity for 32 S. Enteritidis isolates and 100% exclusivity for the 297 non-Enteritidis Salmonella isolates. Single-laboratory validation performed on 1,741 environmental samples demonstrated that the PCR method detected 55% more positives than the Vitek immunodiagnostic assay system (VIDAS) method. The PCR results correlated well with the culture results, and the method did not report any false-negative results. The receiver operating characteristic (ROC) analysis documented excellent agreement between the results from the culture and PCR methods (area under the curve, 0.90; 95% confidence interval of 0.76 to 1.0) confirming the validity of the PCR method. IMPORTANCE This validated PCR method detects 55% more positives for Salmonella in half the time required for the reference method, VIDAS. The validated PCR method will help to strengthen public health efforts through rapid screening of Salmonella spp. in environmental samples. PMID:28500041

76 FR 28781 - Independent Scientific Peer Review Panel Report: Evaluation of the Validation Status of an In...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-05-18

... 29-30, 2011, to evaluate the validation status of the LUMI-CELL[supreg] (BG1Luc ER TA) test method...: http://iccvam.niehs.nih.gov/docs/endo_docs/EDPRPRept2011.pdf or by contacting NICEATM (see ADDRESSES). The report contains (1) the Panel's evaluation of the validation status of the test method and (2) the...

Validation of Diagnostic Measures Based on Latent Class Analysis: A Step Forward in Response Bias Research

ERIC Educational Resources Information Center

Thomas, Michael L.; Lanyon, Richard I.; Millsap, Roger E.

2009-01-01

The use of criterion group validation is hindered by the difficulty of classifying individuals on latent constructs. Latent class analysis (LCA) is a method that can be used for determining the validity of scales meant to assess latent constructs without such a priori classifications. The authors used this method to examine the ability of the L…

Hopes and Cautions for Instrument-Based Evaluation of Consent Capacity: Results of a Construct Validity Study of Three Instruments

PubMed Central

Moye, Jennifer; Azar, Annin R.; Karel, Michele J.; Gurrera, Ronald J.

2016-01-01

Does instrument based evaluation of consent capacity increase the precision and validity of competency assessment or does ostensible precision provide a false sense of confidence without in fact improving validity? In this paper we critically examine the evidence for construct validity of three instruments for measuring four functional abilities important in consent capacity: understanding, appreciation, reasoning, and expressing a choice. Instrument based assessment of these abilities is compared through investigation of a multi-trait multi-method matrix in 88 older adults with mild to moderate dementia. Results find variable support for validity. There appears to be strong evidence for good hetero-method validity for the measurement of understanding, mixed evidence for validity in the measurement of reasoning, and strong evidence for poor hetero-method validity for the concepts of appreciation and expressing a choice, although the latter is likely due to extreme range restrictions. The development of empirically based tools for use in capacity evaluation should ultimately enhance the reliability and validity of assessment, yet clearly more research is needed to define and measure the constructs of decisional capacity. We would also emphasize that instrument based assessment of capacity is only one part of a comprehensive evaluation of competency which includes consideration of diagnosis, psychiatric and/or cognitive symptomatology, risk involved in the situation, and individual and cultural differences. PMID:27330455

Practical Aspects of Designing and Conducting Validation Studies Involving Multi-study Trials.

PubMed

Coecke, Sandra; Bernasconi, Camilla; Bowe, Gerard; Bostroem, Ann-Charlotte; Burton, Julien; Cole, Thomas; Fortaner, Salvador; Gouliarmou, Varvara; Gray, Andrew; Griesinger, Claudius; Louhimies, Susanna; Gyves, Emilio Mendoza-de; Joossens, Elisabeth; Prinz, Maurits-Jan; Milcamps, Anne; Parissis, Nicholaos; Wilk-Zasadna, Iwona; Barroso, João; Desprez, Bertrand; Langezaal, Ingrid; Liska, Roman; Morath, Siegfried; Reina, Vittorio; Zorzoli, Chiara; Zuang, Valérie

This chapter focuses on practical aspects of conducting prospective in vitro validation studies, and in particular, by laboratories that are members of the European Union Network of Laboratories for the Validation of Alternative Methods (EU-NETVAL) that is coordinated by the EU Reference Laboratory for Alternatives to Animal Testing (EURL ECVAM). Prospective validation studies involving EU-NETVAL, comprising a multi-study trial involving several laboratories or "test facilities", typically consist of two main steps: (1) the design of the validation study by EURL ECVAM and (2) the execution of the multi-study trial by a number of qualified laboratories within EU-NETVAL, coordinated and supported by EURL ECVAM. The approach adopted in the conduct of these validation studies adheres to the principles described in the OECD Guidance Document on the Validation and International Acceptance of new or updated test methods for Hazard Assessment No. 34 (OECD 2005). The context and scope of conducting prospective in vitro validation studies is dealt with in Chap. 4 . Here we focus mainly on the processes followed to carry out a prospective validation of in vitro methods involving different laboratories with the ultimate aim of generating a dataset that can support a decision in relation to the possible development of an international test guideline (e.g. by the OECD) or the establishment of performance standards.

Comparison of on-site field measured inorganic arsenic in rice with laboratory measurements using a field deployable method: Method validation.

PubMed

Mlangeni, Angstone Thembachako; Vecchi, Valeria; Norton, Gareth J; Raab, Andrea; Krupp, Eva M; Feldmann, Joerg

2018-10-15

A commercial arsenic field kit designed to measure inorganic arsenic (iAs) in water was modified into a field deployable method (FDM) to measure iAs in rice. While the method has been validated to give precise and accurate results in the laboratory, its on-site field performance has not been evaluated. This study was designed to test the method on-site in Malawi in order to evaluate its accuracy and precision in determination of iAs on-site by comparing with a validated reference method and giving original data on inorganic arsenic in Malawian rice and rice-based products. The method was validated by using the established laboratory-based HPLC-ICPMS. Statistical tests indicated there were no significant differences between on-site and laboratory iAs measurements determined using the FDM (p = 0.263, ά = 0.05) and between on-site measurements and measurements determined using HPLC-ICP-MS (p = 0.299, ά = 0.05). This method allows quick (within 1 h) and efficient screening of rice containing iAs concentrations on-site. Copyright © 2018 Elsevier Ltd. All rights reserved.

A New Green Method for the Quantitative Analysis of Enrofloxacin by Fourier-Transform Infrared Spectroscopy.

PubMed

Rebouças, Camila Tavares; Kogawa, Ana Carolina; Salgado, Hérida Regina Nunes

2018-05-18

Background: A green analytical chemistry method was developed for quantification of enrofloxacin in tablets. The drug, a second-generation fluoroquinolone, was first introduced in veterinary medicine for the treatment of various bacterial species. Objective: This study proposed to develop, validate, and apply a reliable, low-cost, fast, and simple IR spectroscopy method for quantitative routine determination of enrofloxacin in tablets. Methods: The method was completely validated according to the International Conference on Harmonisation guidelines, showing accuracy, precision, selectivity, robustness, and linearity. Results: It was linear over the concentration range of 1.0-3.0 mg with correlation coefficients >0.9999 and LOD and LOQ of 0.12 and 0.36 mg, respectively. Conclusions: Now that this IR method has met performance qualifications, it can be adopted and applied for the analysis of enrofloxacin tablets for production process control. The validated method can also be utilized to quantify enrofloxacin in tablets and thus is an environmentally friendly alternative for the routine analysis of enrofloxacin in quality control. Highlights: A new green method for the quantitative analysis of enrofloxacin by Fourier-Transform Infrared spectroscopy was validated. It is a fast, clean and low-cost alternative for the evaluation of enrofloxacin tablets.

Cross-validation of the Beunen-Malina method to predict adult height.

PubMed

Beunen, Gaston P; Malina, Robert M; Freitas, Duarte I; Maia, José A; Claessens, Albrecht L; Gouveia, Elvio R; Lefevre, Johan

2010-08-01

The purpose of this study was to cross-validate the Beunen-Malina method for non-invasive prediction of adult height. Three hundred and eight boys aged 13, 14, 15 and 16 years from the Madeira Growth Study were observed at annual intervals in 1996, 1997 and 1998 and re-measured 7-8 years later. Height, sitting height and the triceps and subscapular skinfolds were measured; skeletal age was assessed using the Tanner-Whitehouse 2 method. Adult height was measured and predicted using the Beunen-Malina method. Maturity groups were classified using relative skeletal age (skeletal age minus chronological age). Pearson correlations, mean differences and standard errors of estimate (SEE) were calculated. Age-specific correlations between predicted and measured adult height vary between 0.70 and 0.85, while age-specific SEE varies between 3.3 and 4.7 cm. The correlations and SEE are similar to those obtained in the development of the original Beunen-Malina method. The Beunen-Malina method is a valid method to predict adult height in adolescent boys and can be used in European populations or populations from European ancestry. Percentage of predicted adult height is a non-invasive valid method to assess biological maturity.

Finite element analysis of dental implants with validation: to what extent can we expect the model to predict biological phenomena? A literature review and proposal for classification of a validation process.

PubMed

Chang, Yuanhan; Tambe, Abhijit Anil; Maeda, Yoshinobu; Wada, Masahiro; Gonda, Tomoya

2018-03-08

A literature review of finite element analysis (FEA) studies of dental implants with their model validation process was performed to establish the criteria for evaluating validation methods with respect to their similarity to biological behavior. An electronic literature search of PubMed was conducted up to January 2017 using the Medical Subject Headings "dental implants" and "finite element analysis." After accessing the full texts, the context of each article was searched using the words "valid" and "validation" and articles in which these words appeared were read to determine whether they met the inclusion criteria for the review. Of 601 articles published from 1997 to 2016, 48 that met the eligibility criteria were selected. The articles were categorized according to their validation method as follows: in vivo experiments in humans (n = 1) and other animals (n = 3), model experiments (n = 32), others' clinical data and past literature (n = 9), and other software (n = 2). Validation techniques with a high level of sufficiency and efficiency are still rare in FEA studies of dental implants. High-level validation, especially using in vivo experiments tied to an accurate finite element method, needs to become an established part of FEA studies. The recognition of a validation process should be considered when judging the practicality of an FEA study.

Development of a refractive error quality of life scale for Thai adults (the REQ-Thai).

PubMed

Sukhawarn, Roongthip; Wiratchai, Nonglak; Tatsanavivat, Pyatat; Pitiyanuwat, Somwung; Kanato, Manop; Srivannaboon, Sabong; Guyatt, Gordon H

2011-08-01

To develop a scale for measuring refractive error quality of life (QOL) for Thai adults. The full survey comprised 424 respondents from 5 medical centers in Bangkok and from 3 medical centers in Chiangmai, Songkla and KhonKaen provinces. Participants were emmetropes and persons with refractive correction with visual acuity of 20/30 or better An item reduction process was employed by combining 3 methods-expert opinion, impact method and item-total correlation methods. The classical reliability testing and the validity testing including convergent, discriminative and construct validity was performed. The developed questionnaire comprised 87 items in 6 dimensions: 1) quality of vision, 2) visual function, 3) social function, 4) psychological function, 5) symptoms and 6) refractive correction problems. It is the 5-level Likert scale type. The Cronbach's Alpha coefficients of its dimensions ranged from 0.756 to 0. 979. All validity testing were shown to be valid. The construct validity was validated by the confirmatory factor analysis. A short version questionnaire comprised 48 items with good reliability and validity was also developed. This is the first validated instrument for measuring refractive error quality of life for Thai adults that was developed with strong research methodology and large sample size.

Improving the quality of discrete-choice experiments in health: how can we assess validity and reliability?

PubMed

Janssen, Ellen M; Marshall, Deborah A; Hauber, A Brett; Bridges, John F P

2017-12-01

The recent endorsement of discrete-choice experiments (DCEs) and other stated-preference methods by regulatory and health technology assessment (HTA) agencies has placed a greater focus on demonstrating the validity and reliability of preference results. Areas covered: We present a practical overview of tests of validity and reliability that have been applied in the health DCE literature and explore other study qualities of DCEs. From the published literature, we identify a variety of methods to assess the validity and reliability of DCEs. We conceptualize these methods to create a conceptual model with four domains: measurement validity, measurement reliability, choice validity, and choice reliability. Each domain consists of three categories that can be assessed using one to four procedures (for a total of 24 tests). We present how these tests have been applied in the literature and direct readers to applications of these tests in the health DCE literature. Based on a stakeholder engagement exercise, we consider the importance of study characteristics beyond traditional concepts of validity and reliability. Expert commentary: We discuss study design considerations to assess the validity and reliability of a DCE, consider limitations to the current application of tests, and discuss future work to consider the quality of DCEs in healthcare.

JaCVAM-organized international validation study of the in vivo rodent alkaline comet assay for detection of genotoxic carcinogens: II. Summary of definitive validation study results.

PubMed

Uno, Yoshifumi; Kojima, Hajime; Omori, Takashi; Corvi, Raffaella; Honma, Masamistu; Schechtman, Leonard M; Tice, Raymond R; Beevers, Carol; De Boeck, Marlies; Burlinson, Brian; Hobbs, Cheryl A; Kitamoto, Sachiko; Kraynak, Andrew R; McNamee, James; Nakagawa, Yuzuki; Pant, Kamala; Plappert-Helbig, Ulla; Priestley, Catherine; Takasawa, Hironao; Wada, Kunio; Wirnitzer, Uta; Asano, Norihide; Escobar, Patricia A; Lovell, David; Morita, Takeshi; Nakajima, Madoka; Ohno, Yasuo; Hayashi, Makoto

2015-07-01

The in vivo rodent alkaline comet assay (comet assay) is used internationally to investigate the in vivo genotoxic potential of test chemicals. This assay, however, has not previously been formally validated. The Japanese Center for the Validation of Alternative Methods (JaCVAM), with the cooperation of the U.S. NTP Interagency Center for the Evaluation of Alternative Toxicological Methods (NICEATM)/the Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM), the European Centre for the Validation of Alternative Methods (ECVAM), and the Japanese Environmental Mutagen Society/Mammalian Mutagenesis Study Group (JEMS/MMS), organized an international validation study to evaluate the reliability and relevance of the assay for identifying genotoxic carcinogens, using liver and stomach as target organs. The ultimate goal of this exercise was to establish an Organisation for Economic Co-operation and Development (OECD) test guideline. The study protocol was optimized in the pre-validation studies, and then the definitive (4th phase) validation study was conducted in two steps. In the 1st step, assay reproducibility was confirmed among laboratories using four coded reference chemicals and the positive control ethyl methanesulfonate. In the 2nd step, the predictive capability was investigated using 40 coded chemicals with known genotoxic and carcinogenic activity (i.e., genotoxic carcinogens, genotoxic non-carcinogens, non-genotoxic carcinogens, and non-genotoxic non-carcinogens). Based on the results obtained, the in vivo comet assay is concluded to be highly capable of identifying genotoxic chemicals and therefore can serve as a reliable predictor of rodent carcinogenicity. Copyright © 2015 Elsevier B.V. All rights reserved.

Validation of a spectrophotometric assay method for bisoprolol using picric acid.

PubMed

Panainte, Alina-Diana; Bibire, Nela; Tântaru, Gladiola; Apostu, M; Vieriu, Mădălina

2013-01-01

Bisoprolol is a drug belonging to beta blockers drugs used primarily for the treatment of cardiovascular diseases. A spectrophotometric method for quantitative determination of bisoprolol was developed based on the formation of a complex combination between bisoprolol and picric acid. The complex combination of bisoprolol and picric acid has a maximum absorbance peak at 420 nm. Optimum working conditions were established and the method was validated. The method presented a good linearity in the concentration range 5-120 microg/ml (regression coefficient r2 = 0.9992). The RSD for the precision of the method was 1.74 and for the intermediate precision 1.43, and recovery values ranged between 98.25-101.48%. The proposed and validated spectrophotometric method for the determination of bisoprolol is simple and cost effective.

The construct validity of session RPE during an intensive camp in young male Karate athletes.

PubMed

Padulo, Johnny; Chaabène, Helmi; Tabben, Montassar; Haddad, Monoem; Gevat, Cecilia; Vando, Stefano; Maurino, Lucio; Chaouachi, Anis; Chamari, Karim

2014-04-01

the aim of this study was to assess the validity of the session rating of perceived exertion (RPE) method and two objective HR-based methods for quantifying karate's training load (TL) in young Karatekas. eleven athletes (age 12.50±1.84 years) participated in this study. The training period/camp was performed on 5 consecutive days with two training session (s) per-day (d). Construct validity of RPE method in young Karate athletes, was studied by correlation analysis between RPE session's training load and both Edwards and Banister's training impulse score' method. significant relationship was found between inter-day (n-11 × d-5 × s-2 = 110) sessions RPE and Edwards (r values from 0.84 to 0.92 p < 0.001) and Banister's (r values from 0.84 to 0.97 p < 0.001), respectively. this study showed that session-RPE can be considered a valid method for quantifying karate's training load in young karate athletes.

Evaluating the dynamic response of in-flight thrust calculation techniques during throttle transients

NASA Technical Reports Server (NTRS)

Ray, Ronald J.

1994-01-01

New flight test maneuvers and analysis techniques for evaluating the dynamic response of in-flight thrust models during throttle transients have been developed and validated. The approach is based on the aircraft and engine performance relationship between thrust and drag. Two flight test maneuvers, a throttle step and a throttle frequency sweep, were developed and used in the study. Graphical analysis techniques, including a frequency domain analysis method, were also developed and evaluated. They provide quantitative and qualitative results. Four thrust calculation methods were used to demonstrate and validate the test technique. Flight test applications on two high-performance aircraft confirmed the test methods as valid and accurate. These maneuvers and analysis techniques were easy to implement and use. Flight test results indicate the analysis techniques can identify the combined effects of model error and instrumentation response limitations on the calculated thrust value. The methods developed in this report provide an accurate approach for evaluating, validating, or comparing thrust calculation methods for dynamic flight applications.

Validation of newly developed and redesigned key indicator methods for assessment of different working conditions with physical workloads based on mixed-methods design: a study protocol

PubMed Central

Liebers, Falk; Brandstädt, Felix; Schust, Marianne; Serafin, Patrick; Schäfer, Andreas; Gebhardt, Hansjürgen; Hartmann, Bernd; Steinberg, Ulf

2017-01-01

Introduction The impact of work-related musculoskeletal disorders is considerable. The assessment of work tasks with physical workloads is crucial to estimate the work-related health risks of exposed employees. Three key indicator methods are available for risk assessment regarding manual lifting, holding and carrying of loads; manual pulling and pushing of loads; and manual handling operations. Three further KIMs for risk assessment regarding whole-body forces, awkward body postures and body movement have been developed de novo. In addition, the development of a newly drafted combined method for mixed exposures is planned. All methods will be validated regarding face validity, reliability, convergent validity, criterion validity and further aspects of utility under practical conditions. Methods and analysis As part of the joint project MEGAPHYS (multilevel risk assessment of physical workloads), a mixed-methods study is being designed for the validation of KIMs and conducted in companies of different sizes and branches in Germany. Workplaces are documented and analysed by observations, applying KIMs, interviews and assessment of environmental conditions. Furthermore, a survey among the employees at the respective workplaces takes place with standardised questionnaires, interviews and physical examinations. It is intended to include 1200 employees at 120 different workplaces. For analysis of the quality criteria, recommendations of the COSMIN checklist (COnsensus-based Standards for the selection of health Measurement INstruments) will be taken into account. Ethics and dissemination The study was planned and conducted in accordance with the German Medical Professional Code and the Declaration of Helsinki as well as the German Federal Data Protection Act. The design of the study was approved by ethics committees. We intend to publish the validated KIMs in 2018. Results will be published in peer-reviewed journals, presented at international meetings and disseminated to actual users for practical application. PMID:28827239

Objective assessment based on motion-related metrics and technical performance in laparoscopic suturing.

PubMed

Sánchez-Margallo, Juan A; Sánchez-Margallo, Francisco M; Oropesa, Ignacio; Enciso, Silvia; Gómez, Enrique J

2017-02-01

The aim of this study is to present the construct and concurrent validity of a motion-tracking method of laparoscopic instruments based on an optical pose tracker and determine its feasibility as an objective assessment tool of psychomotor skills during laparoscopic suturing. A group of novice ([Formula: see text] laparoscopic procedures), intermediate (11-100 laparoscopic procedures) and experienced ([Formula: see text] laparoscopic procedures) surgeons performed three intracorporeal sutures on an ex vivo porcine stomach. Motion analysis metrics were recorded using the proposed tracking method, which employs an optical pose tracker to determine the laparoscopic instruments' position. Construct validation was measured for all 10 metrics across the three groups and between pairs of groups. Concurrent validation was measured against a previously validated suturing checklist. Checklists were completed by two independent surgeons over blinded video recordings of the task. Eighteen novices, 15 intermediates and 11 experienced surgeons took part in this study. Execution time and path length travelled by the laparoscopic dissector presented construct validity. Experienced surgeons required significantly less time ([Formula: see text]), travelled less distance using both laparoscopic instruments ([Formula: see text]) and made more efficient use of the work space ([Formula: see text]) compared with novice and intermediate surgeons. Concurrent validation showed strong correlation between both the execution time and path length and the checklist score ([Formula: see text] and [Formula: see text], [Formula: see text]). The suturing performance was successfully assessed by the motion analysis method. Construct and concurrent validity of the motion-based assessment method has been demonstrated for the execution time and path length metrics. This study demonstrates the efficacy of the presented method for objective evaluation of psychomotor skills in laparoscopic suturing. However, this method does not take into account the quality of the suture. Thus, future works will focus on developing new methods combining motion analysis and qualitative outcome evaluation to provide a complete performance assessment to trainees.

Comparing Thermal Process Validation Methods for Salmonella Inactivation on Almond Kernels.

PubMed

Jeong, Sanghyup; Marks, Bradley P; James, Michael K

2017-01-01

Ongoing regulatory changes are increasing the need for reliable process validation methods for pathogen reduction processes involving low-moisture products; however, the reliability of various validation methods has not been evaluated. Therefore, the objective was to quantify accuracy and repeatability of four validation methods (two biologically based and two based on time-temperature models) for thermal pasteurization of almonds. Almond kernels were inoculated with Salmonella Enteritidis phage type 30 or Enterococcus faecium (NRRL B-2354) at ~10 8 CFU/g, equilibrated to 0.24, 0.45, 0.58, or 0.78 water activity (a w ), and then heated in a pilot-scale, moist-air impingement oven (dry bulb 121, 149, or 177°C; dew point <33.0, 69.4, 81.6, or 90.6°C; v air = 2.7 m/s) to a target lethality of ~4 log. Almond surface temperatures were measured in two ways, and those temperatures were used to calculate Salmonella inactivation using a traditional (D, z) model and a modified model accounting for process humidity. Among the process validation methods, both methods based on time-temperature models had better repeatability, with replication errors approximately half those of the surrogate ( E. faecium ). Additionally, the modified model yielded the lowest root mean squared error in predicting Salmonella inactivation (1.1 to 1.5 log CFU/g); in contrast, E. faecium yielded a root mean squared error of 1.2 to 1.6 log CFU/g, and the traditional model yielded an unacceptably high error (3.4 to 4.4 log CFU/g). Importantly, the surrogate and modified model both yielded lethality predictions that were statistically equivalent (α = 0.05) to actual Salmonella lethality. The results demonstrate the importance of methodology, a w , and process humidity when validating thermal pasteurization processes for low-moisture foods, which should help processors select and interpret validation methods to ensure product safety.

Display format, highlight validity, and highlight method: Their effects on search performance

NASA Technical Reports Server (NTRS)

Donner, Kimberly A.; Mckay, Tim D.; Obrien, Kevin M.; Rudisill, Marianne

1991-01-01

Display format and highlight validity were shown to affect visual display search performance; however, these studies were conducted on small, artificial displays of alphanumeric stimuli. A study manipulating these variables was conducted using realistic, complex Space Shuttle information displays. A 2x2x3 within-subjects analysis of variance found that search times were faster for items in reformatted displays than for current displays. Responses to valid applications of highlight were significantly faster than responses to non or invalidly highlighted applications. The significant format by highlight validity interaction showed that there was little difference in response time to both current and reformatted displays when the highlight validity was applied; however, under the non or invalid highlight conditions, search times were faster with reformatted displays. A separate within-subject analysis of variance of display format, highlight validity, and several highlight methods did not reveal a main effect of highlight method. In addition, observed display search times were compared to search time predicted by Tullis' Display Analysis Program. Benefits of highlighting and reformatting displays to enhance search and the necessity to consider highlight validity and format characteristics in tandem for predicting search performance are discussed.

Validation conform ISO-15189 of assays in the field of autoimmunity: Joint efforts in The Netherlands.

PubMed

Mulder, Leontine; van der Molen, Renate; Koelman, Carin; van Leeuwen, Ester; Roos, Anja; Damoiseaux, Jan

2018-05-01

ISO 15189:2012 requires validation of methods used in the medical laboratory, and lists a series of performance parameters for consideration to include. Although these performance parameters are feasible for clinical chemistry analytes, application in the validation of autoimmunity tests is a challenge. Lack of gold standards or reference methods in combination with the scarcity of well-defined diagnostic samples of patients with rare diseases make validation of new assays difficult. The present manuscript describes the initiative of Dutch medical immunology laboratory specialists to combine efforts and perform multi-center validation studies of new assays in the field of autoimmunity. Validation data and reports are made available to interested Dutch laboratory specialists. Copyright © 2018 Elsevier B.V. All rights reserved.

Modeling, implementation, and validation of arterial travel time reliability.

DOT National Transportation Integrated Search

2013-11-01

Previous research funded by Florida Department of Transportation (FDOT) developed a method for estimating : travel time reliability for arterials. This method was not initially implemented or validated using field data. This : project evaluated and r...

Development and validation of a stability indicating HPLC method for determination of lisinopril, lisinopril degradation product and parabens in the lisinopril extemporaneous formulation.

PubMed

Beasley, Christopher A; Shaw, Jessica; Zhao, Zack; Reed, Robert A

2005-03-09

The purpose of the research described herein was to develop and validate a stability-indicating HPLC method for lisinopril, lisinopril degradation product (DKP), methyl paraben and propyl paraben in a lisinopril extemporaneous formulation. The method developed in this report is selective for the components listed above, in the presence of the complex and chromatographically rich matrix presented by the Bicitra and Ora-Sweet SF formulation diluents. The method was also shown to have adequate sensitivity with a detection limit of 0.0075 microg/mL (0.03% of lisinopril method concentration). The validation elements investigated showed that the method has acceptable specificity, recovery, linearity, solution stability, and method precision. Acceptable robustness indicates that the assay method remains unaffected by small but deliberate variations, which are described in ICH Q2A and Q2B guidelines.

Quantitative bioanalysis of strontium in human serum by inductively coupled plasma-mass spectrometry

PubMed Central

Somarouthu, Srikanth; Ohh, Jayoung; Shaked, Jonathan; Cunico, Robert L; Yakatan, Gerald; Corritori, Suzana; Tami, Joe; Foehr, Erik D

2015-01-01

Aim: A bioanalytical method using inductively-coupled plasma-mass spectrometry to measure endogenous levels of strontium in human serum was developed and validated. Results & methodology: This article details the experimental procedures used for the method development and validation thus demonstrating the application of the inductively-coupled plasma-mass spectrometry method for quantification of strontium in human serum samples. The assay was validated for specificity, linearity, accuracy, precision, recovery and stability. Significant endogenous levels of strontium are present in human serum samples ranging from 19 to 96 ng/ml with a mean of 34.6 ± 15.2 ng/ml (SD). Discussion & conclusion: Calibration procedures and sample pretreatment were simplified for high throughput analysis. The validation demonstrates that the method was sensitive, selective for quantification of strontium (88Sr) and is suitable for routine clinical testing of strontium in human serum samples. PMID:28031925

Cleaning verification: A five parameter study of a Total Organic Carbon method development and validation for the cleaning assessment of residual detergents in manufacturing equipment.

PubMed

Li, Xue; Ahmad, Imad A Haidar; Tam, James; Wang, Yan; Dao, Gina; Blasko, Andrei

2018-02-05

A Total Organic Carbon (TOC) based analytical method to quantitate trace residues of clean-in-place (CIP) detergents CIP100 ® and CIP200 ® on the surfaces of pharmaceutical manufacturing equipment was developed and validated. Five factors affecting the development and validation of the method were identified: diluent composition, diluent volume, extraction method, location for TOC sample preparation, and oxidant flow rate. Key experimental parameters were optimized to minimize contamination and to improve the sensitivity, recovery, and reliability of the method. The optimized concentration of the phosphoric acid in the swabbing solution was 0.05M, and the optimal volume of the sample solution was 30mL. The swab extraction method was 1min sonication. The use of a clean room, as compared to an isolated lab environment, was not required for method validation. The method was demonstrated to be linear with a correlation coefficient (R) of 0.9999. The average recoveries from stainless steel surfaces at multiple spike levels were >90%. The repeatability and intermediate precision results were ≤5% across the 2.2-6.6ppm range (50-150% of the target maximum carry over, MACO, limit). The method was also shown to be sensitive with a detection limit (DL) of 38ppb and a quantitation limit (QL) of 114ppb. The method validation demonstrated that the developed method is suitable for its intended use. The methodology developed in this study is generally applicable to the cleaning verification of any organic detergents used for the cleaning of pharmaceutical manufacturing equipment made of electropolished stainless steel material. Copyright © 2017 Elsevier B.V. All rights reserved.

Predicting implementation from organizational readiness for change: a study protocol

PubMed Central

2011-01-01

Background There is widespread interest in measuring organizational readiness to implement evidence-based practices in clinical care. However, there are a number of challenges to validating organizational measures, including inferential bias arising from the halo effect and method bias - two threats to validity that, while well-documented by organizational scholars, are often ignored in health services research. We describe a protocol to comprehensively assess the psychometric properties of a previously developed survey, the Organizational Readiness to Change Assessment. Objectives Our objective is to conduct a comprehensive assessment of the psychometric properties of the Organizational Readiness to Change Assessment incorporating methods specifically to address threats from halo effect and method bias. Methods and Design We will conduct three sets of analyses using longitudinal, secondary data from four partner projects, each testing interventions to improve the implementation of an evidence-based clinical practice. Partner projects field the Organizational Readiness to Change Assessment at baseline (n = 208 respondents; 53 facilities), and prospectively assesses the degree to which the evidence-based practice is implemented. We will conduct predictive and concurrent validities using hierarchical linear modeling and multivariate regression, respectively. For predictive validity, the outcome is the change from baseline to follow-up in the use of the evidence-based practice. We will use intra-class correlations derived from hierarchical linear models to assess inter-rater reliability. Two partner projects will also field measures of job satisfaction for convergent and discriminant validity analyses, and will field Organizational Readiness to Change Assessment measures at follow-up for concurrent validity (n = 158 respondents; 33 facilities). Convergent and discriminant validities will test associations between organizational readiness and different aspects of job satisfaction: satisfaction with leadership, which should be highly correlated with readiness, versus satisfaction with salary, which should be less correlated with readiness. Content validity will be assessed using an expert panel and modified Delphi technique. Discussion We propose a comprehensive protocol for validating a survey instrument for assessing organizational readiness to change that specifically addresses key threats of bias related to halo effect, method bias and questions of construct validity that often go unexplored in research using measures of organizational constructs. PMID:21777479

Teaching Analytical Method Transfer through Developing and Validating Then Transferring Dissolution Testing Methods for Pharmaceuticals

ERIC Educational Resources Information Center

Kimaru, Irene; Koether, Marina; Chichester, Kimberly; Eaton, Lafayette

2017-01-01

Analytical method transfer (AMT) and dissolution testing are important topics required in industry that should be taught in analytical chemistry courses. Undergraduate students in senior level analytical chemistry laboratory courses at Kennesaw State University (KSU) and St. John Fisher College (SJFC) participated in development, validation, and…

Validity of the remote food photography method against doubly labeled water among minority preschoolers

USDA-ARS?s Scientific Manuscript database

The aim of this study was to determine the validity of energy intake (EI) estimations made using the remote food photography method (RFPM) compared to the doubly labeled water (DLW) method in minority preschool children in a free-living environment. Seven days of food intake and spot urine samples...

Triangulation, Respondent Validation, and Democratic Participation in Mixed Methods Research

ERIC Educational Resources Information Center

Torrance, Harry

2012-01-01

Over the past 10 years or so the "Field" of "Mixed Methods Research" (MMR) has increasingly been exerting itself as something separate, novel, and significant, with some advocates claiming paradigmatic status. Triangulation is an important component of mixed methods designs. Triangulation has its origins in attempts to validate research findings…

Illustrating a Mixed-Method Approach for Validating Culturally Specific Constructs

ERIC Educational Resources Information Center

Hitchcock, J.H.; Nastasi, B.K.; Dai, D.Y.; Newman, J.; Jayasena, A.; Bernstein-Moore, R.; Sarkar, S.; Varjas, K.

2005-01-01

The purpose of this article is to illustrate a mixed-method approach (i.e., combining qualitative and quantitative methods) for advancing the study of construct validation in cross-cultural research. The article offers a detailed illustration of the approach using the responses 612 Sri Lankan adolescents provided to an ethnographic survey. Such…

Uncertainty estimates of purity measurements based on current information: toward a "live validation" of purity methods.

PubMed

Apostol, Izydor; Kelner, Drew; Jiang, Xinzhao Grace; Huang, Gang; Wypych, Jette; Zhang, Xin; Gastwirt, Jessica; Chen, Kenneth; Fodor, Szilan; Hapuarachchi, Suminda; Meriage, Dave; Ye, Frank; Poppe, Leszek; Szpankowski, Wojciech

2012-12-01

To predict precision and other performance characteristics of chromatographic purity methods, which represent the most widely used form of analysis in the biopharmaceutical industry. We have conducted a comprehensive survey of purity methods, and show that all performance characteristics fall within narrow measurement ranges. This observation was used to develop a model called Uncertainty Based on Current Information (UBCI), which expresses these performance characteristics as a function of the signal and noise levels, hardware specifications, and software settings. We applied the UCBI model to assess the uncertainty of purity measurements, and compared the results to those from conventional qualification. We demonstrated that the UBCI model is suitable to dynamically assess method performance characteristics, based on information extracted from individual chromatograms. The model provides an opportunity for streamlining qualification and validation studies by implementing a "live validation" of test results utilizing UBCI as a concurrent assessment of measurement uncertainty. Therefore, UBCI can potentially mitigate the challenges associated with laborious conventional method validation and facilitates the introduction of more advanced analytical technologies during the method lifecycle.

Development and validation of a discriminating in vitro dissolution method for a poorly soluble drug, olmesartan medoxomil: comparison between commercial tablets.

PubMed

Bajerski, Lisiane; Rossi, Rochele Cassanta; Dias, Carolina Lupi; Bergold, Ana Maria; Fröehlich, Pedro Eduardo

2010-06-01

A dissolution test for tablets containing 40 mg of olmesartan medoxomil (OLM) was developed and validated using both LC-UV and UV methods. After evaluation of the sink condition, dissolution medium, and stability of the drug, the method was validated using USP apparatus 2, 50 rpm rotation speed, and 900 ml of deaerated H(2)O + 0.5% sodium lauryl sulfate (w/v) at pH 6.8 (adjusted with 18% phosphoric acid) as the dissolution medium. The model-independent method using difference factor (f(1)) and similarity factor (f(2)), model-dependent method, and dissolution efficiency were employed to compare dissolution profiles. The kinetic parameters of drug release were also investigated. The obtained results provided adequate dissolution profiles. The developed dissolution test was validated according to international guidelines. Since there is no monograph for this drug in tablets, the dissolution method presented here can be used as a quality control test for OLM in this dosage form, especially in a batch to batch evaluation.

The construct validity of session RPE during an intensive camp in young male Karate athletes

PubMed Central

Padulo, Johnny; Chaabène, Helmi; Tabben, Montassar; Haddad, Monoem; Gevat, Cecilia; Vando, Stefano; Maurino, Lucio; Chaouachi, Anis; Chamari, Karim

2014-01-01

Summary Background: the aim of this study was to assess the validity of the session rating of perceived exertion (RPE) method and two objective HR-based methods for quantifying karate’s training load (TL) in young Karatekas. Methods: eleven athletes (age 12.50±1.84 years) participated in this study. The training period/camp was performed on 5 consecutive days with two training session (s) per-day (d). Construct validity of RPE method in young Karate athletes, was studied by correlation analysis between RPE session’s training load and both Edwards and Banister’s training impulse score’ method. Results: significant relationship was found between inter-day (n-11 × d-5 × s-2 = 110) sessions RPE and Edwards (r values from 0.84 to 0.92 p < 0.001) and Banister’s (r values from 0.84 to 0.97 p < 0.001), respectively Conclusion: this study showed that session-RPE can be considered a valid method for quantifying karate’s training load in young karate athletes. PMID:25332921

Optimization and Validation of a Sensitive Method for HPLC-PDA Simultaneous Determination of Torasemide and Spironolactone in Human Plasma using Central Composite Design.

PubMed

Subramanian, Venkatesan; Nagappan, Kannappan; Sandeep Mannemala, Sai

2015-01-01

A sensitive, accurate, precise and rapid HPLC-PDA method was developed and validated for the simultaneous determination of torasemide and spironolactone in human plasma using Design of experiments. Central composite design was used to optimize the method using content of acetonitrile, concentration of buffer and pH of mobile phase as independent variables, while the retention factor of spironolactone, resolution between torasemide and phenobarbitone; and retention time of phenobarbitone were chosen as dependent variables. The chromatographic separation was achieved on Phenomenex C(18) column and the mobile phase comprising 20 mM potassium dihydrogen ortho phosphate buffer (pH-3.2) and acetonitrile in 82.5:17.5 v/v pumped at a flow rate of 1.0 mL min(-1). The method was validated according to USFDA guidelines in terms of selectivity, linearity, accuracy, precision, recovery and stability. The limit of quantitation values were 80 and 50 ng mL(-1) for torasemide and spironolactone respectively. Furthermore, the sensitivity and simplicity of the method suggests the validity of method for routine clinical studies.

[Validation of measurement methods and estimation of uncertainty of measurement of chemical agents in the air at workstations].

PubMed

Dobecki, Marek

2012-01-01

This paper reviews the requirements for measurement methods of chemical agents in the air at workstations. European standards, which have a status of Polish standards, comprise some requirements and information on sampling strategy, measuring techniques, type of samplers, sampling pumps and methods of occupational exposure evaluation at a given technological process. Measurement methods, including air sampling and analytical procedure in a laboratory, should be appropriately validated before intended use. In the validation process, selected methods are tested and budget of uncertainty is set up. The validation procedure that should be implemented in the laboratory together with suitable statistical tools and major components of uncertainity to be taken into consideration, were presented in this paper. Methods of quality control, including sampling and laboratory analyses were discussed. Relative expanded uncertainty for each measurement expressed as a percentage, should not exceed the limit of values set depending on the type of occupational exposure (short-term or long-term) and the magnitude of exposure to chemical agents in the work environment.

Determination of serum levels of imatinib mesylate in patients with chronic myeloid leukemia: validation and application of a new analytical method to monitor treatment compliance

PubMed Central

Rezende, Vinícius Marcondes; Rivellis, Ariane Julio; Gomes, Melissa Medrano; Dörr, Felipe Augusto; Novaes, Mafalda Megumi Yoshinaga; Nardinelli, Luciana; Costa, Ariel Lais de Lima; Chamone, Dalton de Alencar Fisher; Bendit, Israel

2013-01-01

Objective The goal of this study was to monitor imatinib mesylate therapeutically in the Tumor Biology Laboratory, Department of Hematology and Hemotherapy, Hospital das Clínicas, Faculdade de Medicina, Universidade de São Paulo (USP). A simple and sensitive method to quantify imatinib and its metabolite (CGP74588) in human serum was developed and fully validated in order to monitor treatment compliance. Methods The method used to quantify these compounds in serum included protein precipitation extraction followed by instrumental analysis using high performance liquid chromatography coupled with mass spectrometry. The method was validated for several parameters, including selectivity, precision, accuracy, recovery and linearity. Results The parameters evaluated during the validation stage exhibited satisfactory results based on the Food and Drug Administration and the Brazilian Health Surveillance Agency (ANVISA) guidelines for validating bioanalytical methods. These parameters also showed a linear correlation greater than 0.99 for the concentration range between 0.500 µg/mL and 10.0 µg/mL and a total analysis time of 13 minutes per sample. This study includes results (imatinib serum concentrations) for 308 samples from patients being treated with imatinib mesylate. Conclusion The method developed in this study was successfully validated and is being efficiently used to measure imatinib concentrations in samples from chronic myeloid leukemia patients to check treatment compliance. The imatinib serum levels of patients achieving a major molecular response were significantly higher than those of patients who did not achieve this result. These results are thus consistent with published reports concerning other populations. PMID:23741187

Development and validation of a UPLC method for the determination of duloxetine hydrochloride residues on pharmaceutical manufacturing equipment surfaces

PubMed Central

Kumar, Navneet; Sangeetha, D.; Balakrishna, P.

2011-01-01

Background: In pharmaceutical industries, it is very important to remove drug residues from the equipment and areas used. The cleaning procedure must be validated, so special attention must be devoted to the methods used for analysis of trace amounts of drugs. A rapid, sensitive, and specific reverse phase ultra-performance liquid chromatographic (UPLC) method was developed for the quantitative determination of duloxetine in cleaning validation swab samples. Material and Methods: The method was validated using an Acquity UPLC™ HSS T3 (100 × 2.1 mm2) 1.8 μm column with a isocratic mobile phase containing a mixture of 0.01 M potassium dihydrogen orthophosphate, pH adjusted to 3.0 with orthophosphoric acid and acetonitrile (60:40 v/v). The flow rate of the mobile phase was 0.4 ml/min with a column temperature of 40°C and detection wavelength at 230 nm. Cotton swabs, moisten with extraction solution (90% methanol and 10% water), were used to remove any residue of drug from stainless steel, glass and silica surfaces, and give recoveries >80% at four concentration levels. Results: The precision of the results, reported as the relative standard deviation, were below 1.5%. The calibration curve was linear over a concentration range from 0.02 to 5.0 μg/ml with a correlation coefficient of 0.999. The detection limit and quantitation limit were 0.006 and 0.02 μg/ml, respectively. The method was validated over a concentration range of 0.05–5.0 μg/ml. Conclusion: The developed method was validated with respect to specificity, linearity, limit of detection and quantification, accuracy, precision, and robustness. PMID:23781449

Developmental validation of a Nextera XT mitogenome Illumina MiSeq sequencing method for high-quality samples.

PubMed

Peck, Michelle A; Sturk-Andreaggi, Kimberly; Thomas, Jacqueline T; Oliver, Robert S; Barritt-Ross, Suzanne; Marshall, Charla

2018-05-01

Generating mitochondrial genome (mitogenome) data from reference samples in a rapid and efficient manner is critical to harnessing the greater power of discrimination of the entire mitochondrial DNA (mtDNA) marker. The method of long-range target enrichment, Nextera XT library preparation, and Illumina sequencing on the MiSeq is a well-established technique for generating mitogenome data from high-quality samples. To this end, a validation was conducted for this mitogenome method processing up to 24 samples simultaneously along with analysis in the CLC Genomics Workbench and utilizing the AQME (AFDIL-QIAGEN mtDNA Expert) tool to generate forensic profiles. This validation followed the Federal Bureau of Investigation's Quality Assurance Standards (QAS) for forensic DNA testing laboratories and the Scientific Working Group on DNA Analysis Methods (SWGDAM) validation guidelines. The evaluation of control DNA, non-probative samples, blank controls, mixtures, and nonhuman samples demonstrated the validity of this method. Specifically, the sensitivity was established at ≥25 pg of nuclear DNA input for accurate mitogenome profile generation. Unreproducible low-level variants were observed in samples with low amplicon yields. Further, variant quality was shown to be a useful metric for identifying sequencing error and crosstalk. Success of this method was demonstrated with a variety of reference sample substrates and extract types. These studies further demonstrate the advantages of using NGS techniques by highlighting the quantitative nature of heteroplasmy detection. The results presented herein from more than 175 samples processed in ten sequencing runs, show this mitogenome sequencing method and analysis strategy to be valid for the generation of reference data. Copyright © 2018 Elsevier B.V. All rights reserved.

Optimization and single-laboratory validation of a method for the determination of flavonolignans in milk thistle seeds by high-performance liquid chromatography with ultraviolet detection.

PubMed

Mudge, Elizabeth; Paley, Lori; Schieber, Andreas; Brown, Paula N

2015-10-01

Seeds of milk thistle, Silybum marianum (L.) Gaertn., are used for treatment and prevention of liver disorders and were identified as a high priority ingredient requiring a validated analytical method. An AOAC International expert panel reviewed existing methods and made recommendations concerning method optimization prior to validation. A series of extraction and separation studies were undertaken on the selected method for determining flavonolignans from milk thistle seeds and finished products to address the review panel recommendations. Once optimized, a single-laboratory validation study was conducted. The method was assessed for repeatability, accuracy, selectivity, LOD, LOQ, analyte stability, and linearity. Flavonolignan content ranged from 1.40 to 52.86% in raw materials and dry finished products and ranged from 36.16 to 1570.7 μg/mL in liquid tinctures. Repeatability for the individual flavonolignans in raw materials and finished products ranged from 1.03 to 9.88% RSDr, with HorRat values between 0.21 and 1.55. Calibration curves for all flavonolignan concentrations had correlation coefficients of >99.8%. The LODs for the flavonolignans ranged from 0.20 to 0.48 μg/mL at 288 nm. Based on the results of this single-laboratory validation, this method is suitable for the quantitation of the six major flavonolignans in milk thistle raw materials and finished products, as well as multicomponent products containing dandelion, schizandra berry, and artichoke extracts. It is recommended that this method be adopted as First Action Official Method status by AOAC International.

Evaluating the efficiency of spectral resolution of univariate methods manipulating ratio spectra and comparing to multivariate methods: An application to ternary mixture in common cold preparation

NASA Astrophysics Data System (ADS)

Moustafa, Azza Aziz; Salem, Hesham; Hegazy, Maha; Ali, Omnia

2015-02-01

Simple, accurate, and selective methods have been developed and validated for simultaneous determination of a ternary mixture of Chlorpheniramine maleate (CPM), Pseudoephedrine HCl (PSE) and Ibuprofen (IBF), in tablet dosage form. Four univariate methods manipulating ratio spectra were applied, method A is the double divisor-ratio difference spectrophotometric method (DD-RD). Method B is double divisor-derivative ratio spectrophotometric method (DD-RD). Method C is derivative ratio spectrum-zero crossing method (DRZC), while method D is mean centering of ratio spectra (MCR). Two multivariate methods were also developed and validated, methods E and F are Principal Component Regression (PCR) and Partial Least Squares (PLSs). The proposed methods have the advantage of simultaneous determination of the mentioned drugs without prior separation steps. They were successfully applied to laboratory-prepared mixtures and to commercial pharmaceutical preparation without any interference from additives. The proposed methods were validated according to the ICH guidelines. The obtained results were statistically compared with the official methods where no significant difference was observed regarding both accuracy and precision.

Utility of NIST Whole-Genome Reference Materials for the Technical Validation of a Multigene Next-Generation Sequencing Test.

PubMed

Shum, Bennett O V; Henner, Ilya; Belluoccio, Daniele; Hinchcliffe, Marcus J

2017-07-01

The sensitivity and specificity of next-generation sequencing laboratory developed tests (LDTs) are typically determined by an analyte-specific approach. Analyte-specific validations use disease-specific controls to assess an LDT's ability to detect known pathogenic variants. Alternatively, a methods-based approach can be used for LDT technical validations. Methods-focused validations do not use disease-specific controls but use benchmark reference DNA that contains known variants (benign, variants of unknown significance, and pathogenic) to assess variant calling accuracy of a next-generation sequencing workflow. Recently, four whole-genome reference materials (RMs) from the National Institute of Standards and Technology (NIST) were released to standardize methods-based validations of next-generation sequencing panels across laboratories. We provide a practical method for using NIST RMs to validate multigene panels. We analyzed the utility of RMs in validating a novel newborn screening test that targets 70 genes, called NEO1. Despite the NIST RM variant truth set originating from multiple sequencing platforms, replicates, and library types, we discovered a 5.2% false-negative variant detection rate in the RM truth set genes that were assessed in our validation. We developed a strategy using complementary non-RM controls to demonstrate 99.6% sensitivity of the NEO1 test in detecting variants. Our findings have implications for laboratories or proficiency testing organizations using whole-genome NIST RMs for testing. Copyright © 2017 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved.

Survey Instrument Validity Part I: Principles of Survey Instrument Development and Validation in Athletic Training Education Research

ERIC Educational Resources Information Center

Burton, Laura J.; Mazerolle, Stephanie M.

2011-01-01

Context: Instrument validation is an important facet of survey research methods and athletic trainers must be aware of the important underlying principles. Objective: To discuss the process of survey development and validation, specifically the process of construct validation. Background: Athletic training researchers frequently employ the use of…

Sample Size and Power Estimates for a Confirmatory Factor Analytic Model in Exercise and Sport: A Monte Carlo Approach

ERIC Educational Resources Information Center

Myers, Nicholas D.; Ahn, Soyeon; Jin, Ying

2011-01-01

Monte Carlo methods can be used in data analytic situations (e.g., validity studies) to make decisions about sample size and to estimate power. The purpose of using Monte Carlo methods in a validity study is to improve the methodological approach within a study where the primary focus is on construct validity issues and not on advancing…

An Investigation of Pre-Service Middle School Mathematics Teachers' Ability to Conduct Valid Proofs, Methods Used, and Reasons for Invalid Arguments

ERIC Educational Resources Information Center

Demiray, Esra; Isiksal Bostan, Mine

2017-01-01

The purposes of this study are to investigate Turkish pre-service middle school mathematics teachers' ability in conducting valid proofs for statements regarding numbers and algebra in terms of their year of enrollment in a teacher education program, to determine the proof methods used in their valid proofs, and to examine the reasons for their…

Multiple Imputation based Clustering Validation (MIV) for Big Longitudinal Trial Data with Missing Values in eHealth.

PubMed

Zhang, Zhaoyang; Fang, Hua; Wang, Honggang

2016-06-01

Web-delivered trials are an important component in eHealth services. These trials, mostly behavior-based, generate big heterogeneous data that are longitudinal, high dimensional with missing values. Unsupervised learning methods have been widely applied in this area, however, validating the optimal number of clusters has been challenging. Built upon our multiple imputation (MI) based fuzzy clustering, MIfuzzy, we proposed a new multiple imputation based validation (MIV) framework and corresponding MIV algorithms for clustering big longitudinal eHealth data with missing values, more generally for fuzzy-logic based clustering methods. Specifically, we detect the optimal number of clusters by auto-searching and -synthesizing a suite of MI-based validation methods and indices, including conventional (bootstrap or cross-validation based) and emerging (modularity-based) validation indices for general clustering methods as well as the specific one (Xie and Beni) for fuzzy clustering. The MIV performance was demonstrated on a big longitudinal dataset from a real web-delivered trial and using simulation. The results indicate MI-based Xie and Beni index for fuzzy-clustering are more appropriate for detecting the optimal number of clusters for such complex data. The MIV concept and algorithms could be easily adapted to different types of clustering that could process big incomplete longitudinal trial data in eHealth services.

Multiple Imputation based Clustering Validation (MIV) for Big Longitudinal Trial Data with Missing Values in eHealth

PubMed Central

Zhang, Zhaoyang; Wang, Honggang

2016-01-01

Web-delivered trials are an important component in eHealth services. These trials, mostly behavior-based, generate big heterogeneous data that are longitudinal, high dimensional with missing values. Unsupervised learning methods have been widely applied in this area, however, validating the optimal number of clusters has been challenging. Built upon our multiple imputation (MI) based fuzzy clustering, MIfuzzy, we proposed a new multiple imputation based validation (MIV) framework and corresponding MIV algorithms for clustering big longitudinal eHealth data with missing values, more generally for fuzzy-logic based clustering methods. Specifically, we detect the optimal number of clusters by auto-searching and -synthesizing a suite of MI-based validation methods and indices, including conventional (bootstrap or cross-validation based) and emerging (modularity-based) validation indices for general clustering methods as well as the specific one (Xie and Beni) for fuzzy clustering. The MIV performance was demonstrated on a big longitudinal dataset from a real web-delivered trial and using simulation. The results indicate MI-based Xie and Beni index for fuzzy-clustering is more appropriate for detecting the optimal number of clusters for such complex data. The MIV concept and algorithms could be easily adapted to different types of clustering that could process big incomplete longitudinal trial data in eHealth services. PMID:27126063

Measuring Adverse Events in Helicopter Emergency Medical Services: Establishing Content Validity

PubMed Central

Patterson, P. Daniel; Lave, Judith R.; Martin-Gill, Christian; Weaver, Matthew D.; Wadas, Richard J.; Arnold, Robert M.; Roth, Ronald N.; Mosesso, Vincent N.; Guyette, Francis X.; Rittenberger, Jon C.; Yealy, Donald M.

2015-01-01

Introduction We sought to create a valid framework for detecting Adverse Events (AEs) in the high-risk setting of Helicopter Emergency Medical Services (HEMS). Methods We assembled a panel of 10 expert clinicians (n=6 emergency medicine physicians and n=4 prehospital nurses and flight paramedics) affiliated with a large multi-state HEMS organization in the Northeast U.S. We used a modified Delphi technique to develop a framework for detecting AEs associated with the treatment of critically ill or injured patients. We used a widely applied measure, the Content Validity Index (CVI), to quantify the validity of the framework’s content. Results The expert panel of 10 clinicians reached consensus on a common AE definition and four-step protocol/process for AE detection in HEMS. The consensus-based framework is composed of three main components: 1) a trigger tool, 2) a method for rating proximal cause, and 3) a method for rating AE severity. The CVI findings isolate components of the framework considered content valid. Conclusions We demonstrate a standardized process for the development of a content valid framework for AE detection. The framework is a model for the development of a method for AE identification in other settings, including ground-based EMS. PMID:24003951

PRN 96-1: Tolerance Enforcement Methods - Independent Laboratory Validation by Petitioner

EPA Pesticide Factsheets

This notice is intended to clarify the requirements for submission of an Independent Laboratory Validation to accompany new pesticide analytical methods and does not contain additional data requirements.This notice supersedes PR Notice 88-5.

Testing Standard Reliability Criteria

ERIC Educational Resources Information Center

Sherry, David

2017-01-01

Maul's paper, "Rethinking Traditional Methods of Survey Validation" (Andrew Maul), contains two stages. First he presents empirical results that cast doubt on traditional methods for validating psychological measurement instruments. These results motivate the second stage, a critique of current conceptions of psychological measurement…

Contemporary Test Validity in Theory and Practice: A Primer for Discipline-Based Education Researchers

PubMed Central

Reeves, Todd D.; Marbach-Ad, Gili

2016-01-01

Most discipline-based education researchers (DBERs) were formally trained in the methods of scientific disciplines such as biology, chemistry, and physics, rather than social science disciplines such as psychology and education. As a result, DBERs may have never taken specific courses in the social science research methodology—either quantitative or qualitative—on which their scholarship often relies so heavily. One particular aspect of (quantitative) social science research that differs markedly from disciplines such as biology and chemistry is the instrumentation used to quantify phenomena. In response, this Research Methods essay offers a contemporary social science perspective on test validity and the validation process. The instructional piece explores the concepts of test validity, the validation process, validity evidence, and key threats to validity. The essay also includes an in-depth example of a validity argument and validation approach for a test of student argument analysis. In addition to DBERs, this essay should benefit practitioners (e.g., lab directors, faculty members) in the development, evaluation, and/or selection of instruments for their work assessing students or evaluating pedagogical innovations. PMID:26903498

Validation and use of three complementary analytical methods (LC-FLS, LC-MS/MS and ICP-MS) to evaluate the pharmacokinetics, biodistribution and stability of motexafin gadolinium in plasma and tissues.

PubMed

Miles, Dale R; Mesfin, Mimi; Mody, Tarak D; Stiles, Mark; Lee, Jean; Fiene, John; Denis, Bernie; Boswell, Garry W

2006-05-01

Liquid chromatography-fluorescence (LC-FLS), liquid chromatography-tandem mass spectrometry (LC-MS/MS) and inductively coupled plasma-mass spectrometry (ICP-MS) methods were developed and validated for the evaluation of motexafin gadolinium (MGd, Xcytrin) pharmacokinetics and biodistribution in plasma and tissues. The LC-FLS method exhibited the greatest sensitivity (0.0057 microg mL(-1)), and was used for pharmacokinetic, biodistribution, and protein binding studies with small sample sizes or low MGd concentrations. The LC-MS/MS method, which exhibited a short run time and excellent selectivity, was used for routine clinical plasma sample analysis. The ICP-MS method, which measured total Gd, was used in conjunction with LC methods to assess MGd stability in plasma. All three methods were validated using human plasma. The LC-FLS method was also validated using plasma, liver and kidneys from mice and rats. All three methods were shown to be accurate, precise and robust for each matrix validated. For three mice, the mean (standard deviation) concentration of MGd in plasma/tissues taken 5 hr after dosing with 23 mg kg(-1) MGd was determined by LC-FLS as follows: plasma (0.025+/-0.002 microg mL(-1)), liver (2.89+/-0.45 microg g(-1)), and kidney (6.09+/-1.05 microg g(-1)). Plasma samples from a subset of patients with brain metastases from extracranial tumors were analyzed using both LC-MS/MS and ICP-MS methods. For a representative patient, > or = 90% of the total Gd in plasma was accounted for as MGd over the first hour post dosing. By 24 hr post dosing, 63% of total Gd was accounted for as MGd, indicating some metabolism of MGd.

Development and validation of stability indicating HPLC methods for related substances and assay analyses of amoxicillin and potassium clavulanate mixtures.

PubMed

Bellur Atici, Esen; Yazar, Yücel; Ağtaş, Çağan; Ridvanoğlu, Nurten; Karlığa, Bekir

2017-03-20

Antibacterial combinations consisting of the semisynthetic antibiotic amoxicillin (amox) and the β-lactamase inhibitor potassium clavulanate (clav) are commonly used and several chromatographic methods were reported for their quantification in mixtures. In the present work, single HPLC method for related substances analyses of amoxicillin and potassium clavulanate mixtures was developed and validated according to international conference on harmonization (ICH) guidelines. Eighteen amoxicillin and six potassium clavulanate impurities were successfully separated from each other by using triple gradient elution using a Thermo Hypersil Zorbax BDS C18 (250 mm×4.6mm, 3μm) column with 50μL injection volumes at a wavelength of 215nm. Commercially unavailable impurities were formed by degradation of amoxicillin and potassium clavulanate, identified by LC-MS studies and used during analytical method development and validation studies. Also, process related amoxicillin impurity-P was synthesized and characterized by using nuclear magnetic resonance (NMR) and mass spectroscopy (MS) for the first time. As complementary of this work, an assay method for amoxicillin and potassium clavulanate mixtures was developed and validated; stress-testing and stability studies of amox/clav mixtures was carried out under specified conditions according to ICH and analyzed by using validated stability-indicating assay and related substances methods. Copyright © 2016 Elsevier B.V. All rights reserved.

Postcraniometric sex and ancestry estimation in South Africa: a validation study.

PubMed

Liebenberg, Leandi; Krüger, Gabriele C; L'Abbé, Ericka N; Stull, Kyra E

2018-05-24

With the acceptance of the Daubert criteria as the standards for best practice in forensic anthropological research, more emphasis is being placed on the validation of published methods. Methods, both traditional and novel, need to be validated, adjusted, and refined for optimal performance within forensic anthropological analyses. Recently, a custom postcranial database of modern South Africans was created for use in Fordisc 3.1. Classification accuracies of up to 85% for ancestry estimation and 98% for sex estimation were achieved using a multivariate approach. To measure the external validity and report more realistic performance statistics, an independent sample was tested. The postcrania from 180 black, white, and colored South Africans were measured and classified using the custom postcranial database. A decrease in accuracy was observed for both ancestry estimation (79%) and sex estimation (95%) of the validation sample. When incorporating both sex and ancestry simultaneously, the method achieved 70% accuracy, and 79% accuracy when sex-specific ancestry analyses were run. Classification matrices revealed that postcrania were more likely to misclassify as a result of ancestry rather than sex. While both sex and ancestry influence the size of an individual, sex differences are more marked in the postcranial skeleton and are therefore easier to identify. The external validity of the postcranial database was verified and therefore shown to be a useful tool for forensic casework in South Africa. While the classification rates were slightly lower than the original method, this is expected when a method is generalized.

Validation of a multi-residue method for the determination of several antibiotic groups in honey by LC-MS/MS.

PubMed

Bohm, Detlef A; Stachel, Carolin S; Gowik, Petra

2012-07-01

The presented multi-method was developed for the confirmation of 37 antibiotic substances from the six antibiotic groups: macrolides, lincosamides, quinolones, tetracyclines, pleuromutilines and diamino-pyrimidine derivatives. All substances were analysed simultaneously in a single analytical run with the same procedure, including an extraction with buffer, a clean-up by solid-phase extraction, and the measurement by liquid chromatography tandem mass spectrometry in ESI+ mode. The method was validated on the basis of an in-house validation concept with factorial design by combination of seven factors to check the robustness in a concentration range of 5-50 μg kg(-1). The honeys used were of different types with regard to colour and origin. The values calculated for the validation parameters-decision limit CCα (range, 7.5-12.9 μg kg(-1)), detection capability CCβ (range, 9.4-19.9 μg kg(-1)), within-laboratory reproducibility RSD(wR) (<20% except for tulathromycin with 23.5% and tylvalosin with 21.4 %), repeatability RSD(r) (<20% except for tylvalosin with 21.1%), and recovery (range, 92-106%)-were acceptable and in agreement with the criteria of Commission Decision 2002/657/EC. The validation results showed that the method was applicable for the residue analysis of antibiotics in honey to substances with and without recommended concentrations, although some changes had been tested during validation to determine the robustness of the method.

Initial interlaboratory validation of an analytical method for the determination of lead in canned tuna to be used for monitoring and regulatory purposes.

PubMed

Santiago, E C; Bello, F B B

2003-06-01

The Association of Official Analytical Chemists (AOAC) Standard Method 972.23 (dry ashing and flame atomic absorption spectrophotometry (FAAS)), applied to the analysis of lead in tuna, was validated in three selected local laboratories to determine the acceptability of the method to both the Codex Alimentarius Commission (Codex) and the European Union (EU) Commission for monitoring lead in canned tuna. Initial validation showed that the standard AOAC method as performed in the three participating laboratories cannot satisfy the Codex/EU proposed criteria for the method detection limit for monitoring lead in fish at the present regulation level of 0.5 mg x kg(-1). Modification of the standard method by chelation/concentration of the digest solution before FAAS analysis showed that the modified method has the potential to meet Codex/EU criteria on sensitivity, accuracy and precision at the specified regulation level.

Reliability and Validity of the Footprint Assessment Method Using Photoshop CS5 Software in Young People with Down Syndrome.

PubMed

Gutiérrez-Vilahú, Lourdes; Massó-Ortigosa, Núria; Rey-Abella, Ferran; Costa-Tutusaus, Lluís; Guerra-Balic, Myriam

2016-05-01

People with Down syndrome present skeletal abnormalities in their feet that can be analyzed by commonly used gold standard indices (the Hernández-Corvo index, the Chippaux-Smirak index, the Staheli arch index, and the Clarke angle) based on footprint measurements. The use of Photoshop CS5 software (Adobe Systems Software Ireland Ltd, Dublin, Ireland) to measure footprints has been validated in the general population. The present study aimed to assess the reliability and validity of this footprint assessment technique in the population with Down syndrome. Using optical podography and photography, 44 footprints from 22 patients with Down syndrome (11 men [mean ± SD age, 23.82 ± 3.12 years] and 11 women [mean ± SD age, 24.82 ± 6.81 years]) were recorded in a static bipedal standing position. A blinded observer performed the measurements using a validated manual method three times during the 4-month study, with 2 months between measurements. Test-retest was used to check the reliability of the Photoshop CS5 software measurements. Validity and reliability were obtained by intraclass correlation coefficient (ICC). The reliability test for all of the indices showed very good values for the Photoshop CS5 method (ICC, 0.982-0.995). Validity testing also found no differences between the techniques (ICC, 0.988-0.999). The Photoshop CS5 software method is reliable and valid for the study of footprints in young people with Down syndrome.

[Selection of risk and diagnosis in diabetic polyneuropathy. Validation of method of new systems].

PubMed

Jurado, Jerónimo; Caula, Jacinto; Pou i Torelló, Josep Maria

2006-06-30

In a previous study we developed a specific algorithm, the polyneuropathy selection method (PSM) with 4 parameters (age, HDL-C, HbA1c, and retinopathy), to select patients at risk of diabetic polyneuropathy (DPN). We also developed a simplified method for DPN diagnosis: outpatient polyneuropathy diagnosis (OPD), with 4 variables (symptoms and 3 objective tests). To confirm the validity of conventional tests for DPN diagnosis; to validate the discriminatory power of the PSM and the diagnostic value of OPD by evaluating their relationship to electrodiagnosis studies and objective clinical neurological assessment; and to evaluate the correlation of DPN and pro-inflammatory status. Cross-sectional, crossed association for PSM validation. Paired samples for OPD validation. Primary care in 3 counties. Random sample of 75 subjects from the type-2 diabetes census for PSM evaluation. Thirty DPN patients and 30 non-DPN patients (from 2 DM2 sub-groups in our earlier study) for OPD evaluation. The gold standard for DPN diagnosis will be studied by means of a clinical neurological study (symptoms, physical examination, and sensitivity tests) and electrodiagnosis studies (sensitivity and motor EMG). Risks of neuropathy, macroangiopathy and pro-inflammatory status (PCR, TNF soluble fraction and total TGF-beta1) will be studied in every subject. Electrodiagnosis studies should confirm the validity of conventional tests for DPN diagnosis. PSM and OPD will be valid methods for selecting patients at risk and diagnosing DPN. There will be a significant relationship between DPN and pro-inflammatory tests.

Cross-validation to select Bayesian hierarchical models in phylogenetics.

PubMed

Duchêne, Sebastián; Duchêne, David A; Di Giallonardo, Francesca; Eden, John-Sebastian; Geoghegan, Jemma L; Holt, Kathryn E; Ho, Simon Y W; Holmes, Edward C

2016-05-26

Recent developments in Bayesian phylogenetic models have increased the range of inferences that can be drawn from molecular sequence data. Accordingly, model selection has become an important component of phylogenetic analysis. Methods of model selection generally consider the likelihood of the data under the model in question. In the context of Bayesian phylogenetics, the most common approach involves estimating the marginal likelihood, which is typically done by integrating the likelihood across model parameters, weighted by the prior. Although this method is accurate, it is sensitive to the presence of improper priors. We explored an alternative approach based on cross-validation that is widely used in evolutionary analysis. This involves comparing models according to their predictive performance. We analysed simulated data and a range of viral and bacterial data sets using a cross-validation approach to compare a variety of molecular clock and demographic models. Our results show that cross-validation can be effective in distinguishing between strict- and relaxed-clock models and in identifying demographic models that allow growth in population size over time. In most of our empirical data analyses, the model selected using cross-validation was able to match that selected using marginal-likelihood estimation. The accuracy of cross-validation appears to improve with longer sequence data, particularly when distinguishing between relaxed-clock models. Cross-validation is a useful method for Bayesian phylogenetic model selection. This method can be readily implemented even when considering complex models where selecting an appropriate prior for all parameters may be difficult.

Reliability and Validity of the Footprint Assessment Method Using Photoshop CS5 Software.

PubMed

Gutiérrez-Vilahú, Lourdes; Massó-Ortigosa, Núria; Costa-Tutusaus, Lluís; Guerra-Balic, Myriam

2015-05-01

Several sophisticated methods of footprint analysis currently exist. However, it is sometimes useful to apply standard measurement methods of recognized evidence with an easy and quick application. We sought to assess the reliability and validity of a new method of footprint assessment in a healthy population using Photoshop CS5 software (Adobe Systems Inc, San Jose, California). Forty-two footprints, corresponding to 21 healthy individuals (11 men with a mean ± SD age of 20.45 ± 2.16 years and 10 women with a mean ± SD age of 20.00 ± 1.70 years) were analyzed. Footprints were recorded in static bipedal standing position using optical podography and digital photography. Three trials for each participant were performed. The Hernández-Corvo, Chippaux-Smirak, and Staheli indices and the Clarke angle were calculated by manual method and by computerized method using Photoshop CS5 software. Test-retest was used to determine reliability. Validity was obtained by intraclass correlation coefficient (ICC). The reliability test for all of the indices showed high values (ICC, 0.98-0.99). Moreover, the validity test clearly showed no difference between techniques (ICC, 0.99-1). The reliability and validity of a method to measure, assess, and record the podometric indices using Photoshop CS5 software has been demonstrated. This provides a quick and accurate tool useful for the digital recording of morphostatic foot study parameters and their control.

Proposal for risk-based scientific approach on full and partial validation for general changes in bioanalytical method.

PubMed

Mochizuki, Ayumi; Ieki, Katsunori; Kamimori, Hiroshi; Nagao, Akemi; Nakai, Keiko; Nakayama, Akira; Nanba, Eitaro

2018-04-01

The guidance and several guidelines on bioanalytical method validation, which were issued by the US FDA, EMA and Ministry of Health, Labour and Welfare, list the 'full' validation parameters; however, none of these provide any details for 'partial' validation. Japan Bioanalysis Forum approved a total of three annual discussion groups from 2012 to 2014. In the discussion groups, members from pharmaceutical companies and contract research organizations discussed the details of partial validation from a risk assessment viewpoint based on surveys focusing on bioanalysis of small molecules using LC-MS/MS in Japan. This manuscript presents perspectives and recommendations for most conceivable changes that can be made to full and partial validations by members of the discussion groups based on their experiences and discussions at the Japan Bioanalysis Forum Symposium.

A Tale of Two Methods: Chart and Interview Methods for Identifying Delirium

PubMed Central

Saczynski, Jane S.; Kosar, Cyrus M.; Xu, Guoquan; Puelle, Margaret R.; Schmitt, Eva; Jones, Richard N.; Marcantonio, Edward R.; Wong, Bonnie; Isaza, Ilean; Inouye, Sharon K.

2014-01-01

Background Interview and chart-based methods for identifying delirium have been validated. However, relative strengths and limitations of each method have not been described, nor has a combined approach (using both interviews and chart), been systematically examined. Objectives To compare chart and interview-based methods for identification of delirium. Design, Setting and Participants Participants were 300 patients aged 70+ undergoing major elective surgery (majority were orthopedic surgery) interviewed daily during hospitalization for delirium using the Confusion Assessment Method (CAM; interview-based method) and whose medical charts were reviewed for delirium using a validated chart-review method (chart-based method). We examined rate of agreement on the two methods and patient characteristics of those identified using each approach. Predictive validity for clinical outcomes (length of stay, postoperative complications, discharge disposition) was compared. In the absence of a gold-standard, predictive value could not be calculated. Results The cumulative incidence of delirium was 23% (n= 68) by the interview-based method, 12% (n=35) by the chart-based method and 27% (n=82) by the combined approach. Overall agreement was 80%; kappa was 0.30. The methods differed in detection of psychomotor features and time of onset. The chart-based method missed delirium in CAM-identified patients laacking features of psychomotor agitation or inappropriate behavior. The CAM-based method missed chart-identified cases occurring during the night shift. The combined method had high predictive validity for all clinical outcomes. Conclusions Interview and chart-based methods have specific strengths for identification of delirium. A combined approach captures the largest number and the broadest range of delirium cases. PMID:24512042

Development and Validation of a Method for Determining Tridimensional Angular Displacements with Special Applications to Ice Hockey Motions.

ERIC Educational Resources Information Center

Gagnon, Micheline; And Others

1983-01-01

A method for determining the tridimensional angular displacement of skates during the two-legged stop in ice hockey was developed and validated. The angles were measured by geometry, using a cinecamera and specially equipped skates. The method provides a new tool for kinetic analyses of skating movements. (Authors/PP)

Determination of Trace Elements in Uranium by HPLC-ID-ICP-MS: NTNFC Final Report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Manard, Benjamin Thomas; Wylie, Ernest Miller II; Xu, Ning

This report covers the FY 16 effort for the HPLC-ID-ICP-MS methodology 1) sub-method validation for the group I&II elements, 2) sub-method stood-up and validation for REE, 3) sub-method development for the transition element, and 4) completion of a comprehensive SOP for three families of elements.

Chemometric and biological validation of a capillary electrophoresis metabolomic experiment of Schistosoma mansoni infection in mice.

PubMed

Garcia-Perez, Isabel; Angulo, Santiago; Utzinger, Jürg; Holmes, Elaine; Legido-Quigley, Cristina; Barbas, Coral

2010-07-01

Metabonomic and metabolomic studies are increasingly utilized for biomarker identification in different fields, including biology of infection. The confluence of improved analytical platforms and the availability of powerful multivariate analysis software have rendered the multiparameter profiles generated by these omics platforms a user-friendly alternative to the established analysis methods where the quality and practice of a procedure is well defined. However, unlike traditional assays, validation methods for these new multivariate profiling tools have yet to be established. We propose a validation for models obtained by CE fingerprinting of urine from mice infected with the blood fluke Schistosoma mansoni. We have analysed urine samples from two sets of mice infected in an inter-laboratory experiment where different infection methods and animal husbandry procedures were employed in order to establish the core biological response to a S. mansoni infection. CE data were analysed using principal component analysis. Validation of the scores consisted of permutation scrambling (100 repetitions) and a manual validation method, using a third of the samples (not included in the model) as a test or prediction set. The validation yielded 100% specificity and 100% sensitivity, demonstrating the robustness of these models with respect to deciphering metabolic perturbations in the mouse due to a S. mansoni infection. A total of 20 metabolites across the two experiments were identified that significantly discriminated between S. mansoni-infected and noninfected control samples. Only one of these metabolites, allantoin, was identified as manifesting different behaviour in the two experiments. This study shows the reproducibility of CE-based metabolic profiling methods for disease characterization and screening and highlights the importance of much needed validation strategies in the emerging field of metabolomics.

Consensus methods: review of original methods and their main alternatives used in public health

PubMed Central

Bourrée, Fanny; Michel, Philippe; Salmi, Louis Rachid

2008-01-01

Summary Background Consensus-based studies are increasingly used as decision-making methods, for they have lower production cost than other methods (observation, experimentation, modelling) and provide results more rapidly. The objective of this paper is to describe the principles and methods of the four main methods, Delphi, nominal group, consensus development conference and RAND/UCLA, their use as it appears in peer-reviewed publications and validation studies published in the healthcare literature. Methods A bibliographic search was performed in Pubmed/MEDLINE, Banque de Données Santé Publique (BDSP), The Cochrane Library, Pascal and Francis. Keywords, headings and qualifiers corresponding to a list of terms and expressions related to the consensus methods were searched in the thesauri, and used in the literature search. A search with the same terms and expressions was performed on Internet using the website Google Scholar. Results All methods, precisely described in the literature, are based on common basic principles such as definition of subject, selection of experts, and direct or remote interaction processes. They sometimes use quantitative assessment for ranking items. Numerous variants of these methods have been described. Few validation studies have been implemented. Not implementing these basic principles and failing to describe the methods used to reach the consensus were both frequent reasons contributing to raise suspicion regarding the validity of consensus methods. Conclusion When it is applied to a new domain with important consequences in terms of decision making, a consensus method should be first validated. PMID:19013039

Verification and Validation Process for Progressive Damage and Failure Analysis Methods in the NASA Advanced Composites Consortium

NASA Technical Reports Server (NTRS)

Wanthal, Steven; Schaefer, Joseph; Justusson, Brian; Hyder, Imran; Engelstad, Stephen; Rose, Cheryl

2017-01-01

The Advanced Composites Consortium is a US Government/Industry partnership supporting technologies to enable timeline and cost reduction in the development of certified composite aerospace structures. A key component of the consortium's approach is the development and validation of improved progressive damage and failure analysis methods for composite structures. These methods will enable increased use of simulations in design trade studies and detailed design development, and thereby enable more targeted physical test programs to validate designs. To accomplish this goal with confidence, a rigorous verification and validation process was developed. The process was used to evaluate analysis methods and associated implementation requirements to ensure calculation accuracy and to gage predictability for composite failure modes of interest. This paper introduces the verification and validation process developed by the consortium during the Phase I effort of the Advanced Composites Project. Specific structural failure modes of interest are first identified, and a subset of standard composite test articles are proposed to interrogate a progressive damage analysis method's ability to predict each failure mode of interest. Test articles are designed to capture the underlying composite material constitutive response as well as the interaction of failure modes representing typical failure patterns observed in aerospace structures.

The effect of adopting new storage methods for extending product validity periods on manufacturers expected inventory costs.

PubMed

Chen, Po-Yu

2014-01-01

The validness of the expiration dates (validity period) that manufacturers provide on food product labels is a crucial food safety problem. Governments must study how to use their authority by implementing fair awards and punishments to prompt manufacturers into adopting rigorous considerations, such as the effect of adopting new storage methods for extending product validity periods on expected costs. Assuming that a manufacturer sells fresh food or drugs, this manufacturer must respond to current stochastic demands at each unit of time to determine the purchase amount of products for sale. If this decision maker is capable and an opportunity arises, new packaging methods (e.g., aluminum foil packaging, vacuum packaging, high-temperature sterilization after glass packaging, or packaging with various degrees of dryness) or storage methods (i.e., adding desiccants or various antioxidants) can be chosen to extend the validity periods of products. To minimize expected costs, this decision maker must be aware of the processing costs of new storage methods, inventory standards, inventory cycle lengths, and changes in relationships between factors such as stochastic demand functions in a cycle. Based on these changes in relationships, this study established a mathematical model as a basis for discussing the aforementioned topics.

Möglichkeiten und Grenzen der Validierung flächenhaft modellierter Nitrateinträge ins Grundwasser mit der N2/Ar-Methode

NASA Astrophysics Data System (ADS)

Eschenbach, Wolfram; Budziak, Dörte; Elbracht, Jörg; Höper, Heinrich; Krienen, Lisa; Kunkel, Ralf; Meyer, Knut; Well, Reinhard; Wendland, Frank

2018-06-01

Valid models for estimating nitrate emissions from agriculture to groundwater are an indispensable forecasting tool. A major challenge for model validation is the spatial and temporal inconsistency between data from groundwater monitoring points and modelled nitrate inputs into groundwater, and the fact that many existing groundwater monitoring wells cannot be used for validation. With the help of the N2/Ar-method, groundwater monitoring wells in areas with reduced groundwater can now be used for model validation. For this purpose, 484 groundwater monitoring wells were sampled in Lower Saxony. For the first time, modelled potential nitrate concentrations in groundwater recharge (from the DENUZ model) were compared with nitrate input concentrations, which were calculated using the N2/Ar method. The results show a good agreement between both methods for glacial outwash plains and moraine deposits. Although the nitrate degradation processes in groundwater and soil merge seamlessly in areas with a shallow groundwater table, the DENUZ model only calculates denitrification in the soil zone. The DENUZ model thus predicts 27% higher nitrate emissions into the groundwater than the N2/Ar method in such areas. To account for high temporal and spatial variability of nitrate emissions into groundwater, a large number of groundwater monitoring points must be investigated for model validation.

Evaluation of biologic occupational risk control practices: quality indicators development and validation.

PubMed

Takahashi, Renata Ferreira; Gryschek, Anna Luíza F P L; Izumi Nichiata, Lúcia Yasuko; Lacerda, Rúbia Aparecida; Ciosak, Suely Itsuko; Gir, Elucir; Padoveze, Maria Clara

2010-05-01

There is growing demand for the adoption of qualification systems for health care practices. This study is aimed at describing the development and validation of indicators for evaluation of biologic occupational risk control programs. The study involved 3 stages: (1) setting up a research team, (2) development of indicators, and (3) validation of the indicators by a team of specialists recruited to validate each attribute of the developed indicators. The content validation method was used for the validation, and a psychometric scale was developed for the specialists' assessment. A consensus technique was used, and every attribute that obtained a Content Validity Index of at least 0.75 was approved. Eight indicators were developed for the evaluation of the biologic occupational risk prevention program, with emphasis on accidents caused by sharp instruments and occupational tuberculosis prevention. The indicators included evaluation of the structure, process, and results at the prevention and biologic risk control levels. The majority of indicators achieved a favorable consensus regarding all validated attributes. The developed indicators were considered validated, and the method used for construction and validation proved to be effective. Copyright (c) 2010 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Mosby, Inc. All rights reserved.

Investigation of a Sybr-Green-Based Method to Validate DNA Sequences for DNA Computing

DTIC Science & Technology

2005-05-01

OF A SYBR-GREEN-BASED METHOD TO VALIDATE DNA SEQUENCES FOR DNA COMPUTING 6. AUTHOR(S) Wendy Pogozelski, Salvatore Priore, Matthew Bernard ...simulated annealing. Biochemistry, 35, 14077-14089. 15 Pogozelski, W.K., Bernard , M.P. and Macula, A. (2004) DNA code validation using...and Clark, B.F.C. (eds) In RNA Biochemistry and Biotechnology, NATO ASI Series, Kluwer Academic Publishers. Zucker, M. and Stiegler , P. (1981

Validating for Use and Interpretation: A Mixed Methods Contribution Illustrated

ERIC Educational Resources Information Center

Morell, Linda; Tan, Rachael Jin Bee

2009-01-01

Researchers in the areas of psychology and education strive to understand the intersections among validity, educational measurement, and cognitive theory. Guided by a mixed model conceptual framework, this study investigates how respondents' opinions inform the validation argument. Validity evidence for a science assessment was collected through…

Novel Automated Morphometric and Kinematic Handwriting Assessment: A Validity Study in Children with ASD and ADHD

ERIC Educational Resources Information Center

Dirlikov, Benjamin; Younes, Laurent; Nebel, Mary Beth; Martinelli, Mary Katherine; Tiedemann, Alyssa Nicole; Koch, Carolyn A.; Fiorilli, Diana; Bastian, Amy J.; Denckla, Martha Bridge; Miller, Michael I.; Mostofsky, Stewart H.

2017-01-01

This study presents construct validity for a novel automated morphometric and kinematic handwriting assessment, including (1) convergent validity, establishing reliability of automated measures with traditional manual-derived Minnesota Handwriting Assessment (MHA), and (2) discriminant validity, establishing that the automated methods distinguish…

Formal verification and testing: An integrated approach to validating Ada programs

NASA Technical Reports Server (NTRS)

Cohen, Norman H.

1986-01-01

An integrated set of tools called a validation environment is proposed to support the validation of Ada programs by a combination of methods. A Modular Ada Validation Environment (MAVEN) is described which proposes a context in which formal verification can fit into the industrial development of Ada software.

Adolescent Domain Screening Inventory-Short Form: Development and Initial Validation

ERIC Educational Resources Information Center

Corrigan, Matthew J.

2017-01-01

This study sought to develop a short version of the ADSI, and investigate its psychometric properties. Methods: This is a secondary analysis. Analysis to determine the Cronbach's Alpha, correlations to determine concurrent criterion validity and known instrument validity and a logistic regression to determine predictive validity were conducted.…

Initial Reliability and Validity of the Perceived Social Competence Scale

ERIC Educational Resources Information Center

Anderson-Butcher, Dawn; Iachini, Aidyn L.; Amorose, Anthony J.

2008-01-01

Objective: This study describes the development and validation of a perceived social competence scale that social workers can easily use to assess children's and youth's social competence. Method: Exploratory and confirmatory factor analyses were conducted on a calibration and a cross-validation sample of youth. Predictive validity was also…

A program needs-driven approach to selecting dietary assessment methods for decision-making in food fortification programs.

PubMed

Coates, Jennifer; Colaiezzi, Brooke; Fiedler, John L; Wirth, James; Lividini, Keith; Rogers, Beatrice

2012-09-01

Dietary assessment data are essential for designing, monitoring, and evaluating food fortification and other food-based nutrition programs. Planners and managers must understand the validity, usefulness, and cost tradeoffs of employing alternative dietary assessment methods, but little guidance exists. To identify and apply criteria to assess the tradeoffs of using alternative dietary methods for meeting fortification programming needs. Twenty-five semistructured expert interviews were conducted and literature was reviewed for information on the validity, usefulness, and cost of using 24-hour recalls, Food Frequency Questionnaires/Fortification Rapid Assessment Tool (FFQ/FRAT), Food Balance Sheets (FBS), and Household Consumption and Expenditures Surveys (HCES) for program stage-specific information needs. Criteria were developed and applied to construct relative rankings of the four methods. Needs assessment: HCES offers the greatest suitability at the lowest cost for estimating the risk of inadequate intakes, but relative to 24-hour recall compromises validity. HCES should be used to identify vehicles and to estimate coverage and likely impact due to its low cost and moderate-to-high validity. Baseline assessment: 24-hour recall should be applied using a representative sample. Monitoring: A simple, low-cost FFQ can be used to monitor coverage. Impact evaluation: 24-hour recall should be used to assess changes in nutrient intakes. FBS have low validity relative to other methods for all programmatic purposes. Each dietary assessment method has strengths and weaknesses that vary by context and purpose. Method selection must be driven by the program's data needs, the suitability of the methods for the purpose, and a clear understanding of the tradeoffs involved.

Validation and Verification (V and V) Testing on Midscale Flame Resistant (FR) Test Method

DTIC Science & Technology

2016-12-16

Method for Evaluation of Flame Resistant Clothing for Protection against Fire Simulations Using an Instrumented Manikin. Validation and...complement (not replace) the capabilities of the ASTM F1930 Standard Test Method for Evaluation of Flame Resistant Clothing for Protection against Fire ...Engineering Center (NSRDEC) to complement the ASTM F1930 Standard Test Method for Evaluation of Flame Resistant Clothing for Protection against Fire

Concept Development for Future Domains: A New Method of Knowledge Elicitation

DTIC Science & Technology

2005-06-01

Procedure: U.S. Army Research Institute for the Behavioral and Social Sciences (ARI) examined methods to generate, refine, test , and validate new...generate, elaborate, refine, describe, test , and validate new Future Force concepts relating to doctrine, tactics, techniques, procedures, unit and team...System (Harvey, 1993), and the Job Element Method (Primoff & Eyde , 1988). Figure 1 provides a more comprehensive list of task analytic methods. Please see

HPLC-MS/MS method for dexmedetomidine quantification with Design of Experiments approach: application to pediatric pharmacokinetic study.

PubMed

Szerkus, Oliwia; Struck-Lewicka, Wiktoria; Kordalewska, Marta; Bartosińska, Ewa; Bujak, Renata; Borsuk, Agnieszka; Bienert, Agnieszka; Bartkowska-Śniatkowska, Alicja; Warzybok, Justyna; Wiczling, Paweł; Nasal, Antoni; Kaliszan, Roman; Markuszewski, Michał Jan; Siluk, Danuta

2017-02-01

The purpose of this work was to develop and validate a rapid and robust LC-MS/MS method for the determination of dexmedetomidine (DEX) in plasma, suitable for analysis of a large number of samples. Systematic approach, Design of Experiments, was applied to optimize ESI source parameters and to evaluate method robustness, therefore, a rapid, stable and cost-effective assay was developed. The method was validated according to US FDA guidelines. LLOQ was determined at 5 pg/ml. The assay was linear over the examined concentration range (5-2500 pg/ml), Results: Experimental design approach was applied for optimization of ESI source parameters and evaluation of method robustness. The method was validated according to the US FDA guidelines. LLOQ was determined at 5 pg/ml. The assay was linear over the examined concentration range (R 2 > 0.98). The accuracies, intra- and interday precisions were less than 15%. The stability data confirmed reliable behavior of DEX under tested conditions. Application of Design of Experiments approach allowed for fast and efficient analytical method development and validation as well as for reduced usage of chemicals necessary for regular method optimization. The proposed technique was applied to determination of DEX pharmacokinetics in pediatric patients undergoing long-term sedation in the intensive care unit.

External model validation of binary clinical risk prediction models in cardiovascular and thoracic surgery.

PubMed

Hickey, Graeme L; Blackstone, Eugene H

2016-08-01

Clinical risk-prediction models serve an important role in healthcare. They are used for clinical decision-making and measuring the performance of healthcare providers. To establish confidence in a model, external model validation is imperative. When designing such an external model validation study, thought must be given to patient selection, risk factor and outcome definitions, missing data, and the transparent reporting of the analysis. In addition, there are a number of statistical methods available for external model validation. Execution of a rigorous external validation study rests in proper study design, application of suitable statistical methods, and transparent reporting. Copyright © 2016 The American Association for Thoracic Surgery. Published by Elsevier Inc. All rights reserved.

Validation of asthma recording in electronic health records: a systematic review

PubMed Central

Nissen, Francis; Quint, Jennifer K; Wilkinson, Samantha; Mullerova, Hana; Smeeth, Liam; Douglas, Ian J

2017-01-01

Objective To describe the methods used to validate asthma diagnoses in electronic health records and summarize the results of the validation studies. Background Electronic health records are increasingly being used for research on asthma to inform health services and health policy. Validation of the recording of asthma diagnoses in electronic health records is essential to use these databases for credible epidemiological asthma research. Methods We searched EMBASE and MEDLINE databases for studies that validated asthma diagnoses detected in electronic health records up to October 2016. Two reviewers independently assessed the full text against the predetermined inclusion criteria. Key data including author, year, data source, case definitions, reference standard, and validation statistics (including sensitivity, specificity, positive predictive value [PPV], and negative predictive value [NPV]) were summarized in two tables. Results Thirteen studies met the inclusion criteria. Most studies demonstrated a high validity using at least one case definition (PPV >80%). Ten studies used a manual validation as the reference standard; each had at least one case definition with a PPV of at least 63%, up to 100%. We also found two studies using a second independent database to validate asthma diagnoses. The PPVs of the best performing case definitions ranged from 46% to 58%. We found one study which used a questionnaire as the reference standard to validate a database case definition; the PPV of the case definition algorithm in this study was 89%. Conclusion Attaining high PPVs (>80%) is possible using each of the discussed validation methods. Identifying asthma cases in electronic health records is possible with high sensitivity, specificity or PPV, by combining multiple data sources, or by focusing on specific test measures. Studies testing a range of case definitions show wide variation in the validity of each definition, suggesting this may be important for obtaining asthma definitions with optimal validity. PMID:29238227

Validation of newly developed and redesigned key indicator methods for assessment of different working conditions with physical workloads based on mixed-methods design: a study protocol.

PubMed

Klussmann, Andre; Liebers, Falk; Brandstädt, Felix; Schust, Marianne; Serafin, Patrick; Schäfer, Andreas; Gebhardt, Hansjürgen; Hartmann, Bernd; Steinberg, Ulf

2017-08-21

The impact of work-related musculoskeletal disorders is considerable. The assessment of work tasks with physical workloads is crucial to estimate the work-related health risks of exposed employees. Three key indicator methods are available for risk assessment regarding manual lifting, holding and carrying of loads; manual pulling and pushing of loads; and manual handling operations. Three further KIMs for risk assessment regarding whole-body forces, awkward body postures and body movement have been developed de novo. In addition, the development of a newly drafted combined method for mixed exposures is planned. All methods will be validated regarding face validity, reliability, convergent validity, criterion validity and further aspects of utility under practical conditions. As part of the joint project MEGAPHYS (multilevel risk assessment of physical workloads), a mixed-methods study is being designed for the validation of KIMs and conducted in companies of different sizes and branches in Germany. Workplaces are documented and analysed by observations, applying KIMs, interviews and assessment of environmental conditions. Furthermore, a survey among the employees at the respective workplaces takes place with standardised questionnaires, interviews and physical examinations. It is intended to include 1200 employees at 120 different workplaces. For analysis of the quality criteria, recommendations of the COSMIN checklist (COnsensus-based Standards for the selection of health Measurement INstruments) will be taken into account. The study was planned and conducted in accordance with the German Medical Professional Code and the Declaration of Helsinki as well as the German Federal Data Protection Act. The design of the study was approved by ethics committees. We intend to publish the validated KIMs in 2018. Results will be published in peer-reviewed journals, presented at international meetings and disseminated to actual users for practical application. © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2017. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

Geographic and temporal validity of prediction models: Different approaches were useful to examine model performance

PubMed Central

Austin, Peter C.; van Klaveren, David; Vergouwe, Yvonne; Nieboer, Daan; Lee, Douglas S.; Steyerberg, Ewout W.

2017-01-01

Objective Validation of clinical prediction models traditionally refers to the assessment of model performance in new patients. We studied different approaches to geographic and temporal validation in the setting of multicenter data from two time periods. Study Design and Setting We illustrated different analytic methods for validation using a sample of 14,857 patients hospitalized with heart failure at 90 hospitals in two distinct time periods. Bootstrap resampling was used to assess internal validity. Meta-analytic methods were used to assess geographic transportability. Each hospital was used once as a validation sample, with the remaining hospitals used for model derivation. Hospital-specific estimates of discrimination (c-statistic) and calibration (calibration intercepts and slopes) were pooled using random effects meta-analysis methods. I2 statistics and prediction interval width quantified geographic transportability. Temporal transportability was assessed using patients from the earlier period for model derivation and patients from the later period for model validation. Results Estimates of reproducibility, pooled hospital-specific performance, and temporal transportability were on average very similar, with c-statistics of 0.75. Between-hospital variation was moderate according to I2 statistics and prediction intervals for c-statistics. Conclusion This study illustrates how performance of prediction models can be assessed in settings with multicenter data at different time periods. PMID:27262237

Validity of self-reported mechanical demands for occupational epidemiologic research of musculoskeletal disorders

PubMed Central

Barrero, Lope H; Katz, Jeffrey N; Dennerlein, Jack T

2012-01-01

Objectives To describe the relation of the measured validity of self-reported mechanical demands (self-reports) with the quality of validity assessments and the variability of the assessed exposure in the study population. Methods We searched for original articles, published between 1990 and 2008, reporting the validity of self-reports in three major databases: EBSCOhost, Web of Science, and PubMed. Identified assessments were classified by methodological characteristics (eg, type of self-report and reference method) and exposure dimension was measured. We also classified assessments by the degree of comparability between the self-report and the employed reference method, and the variability of the assessed exposure in the study population. Finally, we examined the association of the published validity (r) with this degree of comparability, as well as with the variability of the exposure variable in the study population. Results Of the 490 assessments identified, 75% used observation-based reference measures and 55% tested self-reports of posture duration and movement frequency. Frequently, validity studies did not report demographic information (eg, education, age, and gender distribution). Among assessments reporting correlations as a measure of validity, studies with a better match between the self-report and the reference method, and studies conducted in more heterogeneous populations tended to report higher correlations [odds ratio (OR) 2.03, 95% confidence interval (95% CI) 0.89–4.65 and OR 1.60, 95% CI 0.96–2.61, respectively]. Conclusions The reported data support the hypothesis that validity depends on study-specific factors often not examined. Experimentally manipulating the testing setting could lead to a better understanding of the capabilities and limitations of self-reported information. PMID:19562235

Development and Validation of New Discriminative Dissolution Method for Carvedilol Tablets

PubMed Central

Raju, V.; Murthy, K. V. R.

2011-01-01

The objective of the present study was to develop and validate a discriminative dissolution method for evaluation of carvedilol tablets. Different conditions such as type of dissolution medium, volume of dissolution medium and rotation speed of paddle were evaluated. The best in vitro dissolution profile was obtained using Apparatus II (paddle), 50 rpm, 900 ml of pH 6.8 phosphate buffer as dissolution medium. The drug release was evaluated by high-performance liquid chromatographic method. The dissolution method was validated according to current ICH and FDA guidelines using parameters such as the specificity, accuracy, precision and stability were evaluated and obtained results were within the acceptable range. The comparison of the obtained dissolution profiles of three different products were investigated using ANOVA-based, model-dependent and model-independent methods, results showed that there is significant difference between the products. The dissolution test developed and validated was adequate for its higher discriminative capacity in differentiating the release characteristics of the products tested and could be applied for development and quality control of carvedilol tablets. PMID:22923865

Development and Validation of High Performance Liquid Chromatography Method for Determination Atorvastatin in Tablet

NASA Astrophysics Data System (ADS)

Yugatama, A.; Rohmani, S.; Dewangga, A.

2018-03-01

Atorvastatin is the primary choice for dyslipidemia treatment. Due to patent expiration of atorvastatin, the pharmaceutical industry makes copy of the drug. Therefore, the development methods for tablet quality tests involving atorvastatin concentration on tablets needs to be performed. The purpose of this research was to develop and validate the simple atorvastatin tablet analytical method by HPLC. HPLC system used in this experiment consisted of column Cosmosil C18 (150 x 4,6 mm, 5 µm) as the stationary reverse phase chomatography, a mixture of methanol-water at pH 3 (80:20 v/v) as the mobile phase, flow rate of 1 mL/min, and UV detector at wavelength of 245 nm. Validation methods were including: selectivity, linearity, accuracy, precision, limit of detection (LOD), and limit of quantitation (LOQ). The results of this study indicate that the developed method had good validation including selectivity, linearity, accuracy, precision, LOD, and LOQ for analysis of atorvastatin tablet content. LOD and LOQ were 0.2 and 0.7 ng/mL, and the linearity range were 20 - 120 ng/mL.

Center of pressure based segment inertial parameters validation

PubMed Central

Rezzoug, Nasser; Gorce, Philippe; Isableu, Brice; Venture, Gentiane

2017-01-01

By proposing efficient methods for estimating Body Segment Inertial Parameters’ (BSIP) estimation and validating them with a force plate, it is possible to improve the inverse dynamic computations that are necessary in multiple research areas. Until today a variety of studies have been conducted to improve BSIP estimation but to our knowledge a real validation has never been completely successful. In this paper, we propose a validation method using both kinematic and kinetic parameters (contact forces) gathered from optical motion capture system and a force plate respectively. To compare BSIPs, we used the measured contact forces (Force plate) as the ground truth, and reconstructed the displacements of the Center of Pressure (COP) using inverse dynamics from two different estimation techniques. Only minor differences were seen when comparing the estimated segment masses. Their influence on the COP computation however is large and the results show very distinguishable patterns of the COP movements. Improving BSIP techniques is crucial and deviation from the estimations can actually result in large errors. This method could be used as a tool to validate BSIP estimation techniques. An advantage of this approach is that it facilitates the comparison between BSIP estimation methods and more specifically it shows the accuracy of those parameters. PMID:28662090

Interlaboratory study of a liquid chromatography method for erythromycin: determination of uncertainty.

PubMed

Dehouck, P; Vander Heyden, Y; Smeyers-Verbeke, J; Massart, D L; Marini, R D; Chiap, P; Hubert, Ph; Crommen, J; Van de Wauw, W; De Beer, J; Cox, R; Mathieu, G; Reepmeyer, J C; Voigt, B; Estevenon, O; Nicolas, A; Van Schepdael, A; Adams, E; Hoogmartens, J

2003-08-22

Erythromycin is a mixture of macrolide antibiotics produced by Saccharopolyspora erythreas during fermentation. A new method for the analysis of erythromycin by liquid chromatography has previously been developed. It makes use of an Astec C18 polymeric column. After validation in one laboratory, the method was now validated in an interlaboratory study. Validation studies are commonly used to test the fitness of the analytical method prior to its use for routine quality testing. The data derived in the interlaboratory study can be used to make an uncertainty statement as well. The relationship between validation and uncertainty statement is not clear for many analysts and there is a need to show how the existing data, derived during validation, can be used in practice. Eight laboratories participated in this interlaboratory study. The set-up allowed the determination of the repeatability variance, s(2)r and the between-laboratory variance, s(2)L. Combination of s(2)r and s(2)L results in the reproducibility variance s(2)R. It has been shown how these data can be used in future by a single laboratory that wants to make an uncertainty statement concerning the same analysis.

On the Simulation of Sea States with High Significant Wave Height for the Validation of Parameter Retrieval Algorithms for Future Altimetry Missions

NASA Astrophysics Data System (ADS)

Kuschenerus, Mieke; Cullen, Robert

2016-08-01

To ensure reliability and precision of wave height estimates for future satellite altimetry missions such as Sentinel 6, reliable parameter retrieval algorithms that can extract significant wave heights up to 20 m have to be established. The retrieved parameters, i.e. the retrieval methods need to be validated extensively on a wide range of possible significant wave heights. Although current missions require wave height retrievals up to 20 m, there is little evidence of systematic validation of parameter retrieval methods for sea states with wave heights above 10 m. This paper provides a definition of a set of simulated sea states with significant wave height up to 20 m, that allow simulation of radar altimeter response echoes for extreme sea states in SAR and low resolution mode. The simulated radar responses are used to derive significant wave height estimates, which can be compared with the initial models, allowing precision estimations of the applied parameter retrieval methods. Thus we establish a validation method for significant wave height retrieval for sea states causing high significant wave heights, to allow improved understanding and planning of future satellite altimetry mission validation.

Validating Remotely Sensed Land Surface Evapotranspiration Based on Multi-scale Field Measurements

NASA Astrophysics Data System (ADS)

Jia, Z.; Liu, S.; Ziwei, X.; Liang, S.

2012-12-01

The land surface evapotranspiration plays an important role in the surface energy balance and the water cycle. There have been significant technical and theoretical advances in our knowledge of evapotranspiration over the past two decades. Acquisition of the temporally and spatially continuous distribution of evapotranspiration using remote sensing technology has attracted the widespread attention of researchers and managers. However, remote sensing technology still has many uncertainties coming from model mechanism, model inputs, parameterization schemes, and scaling issue in the regional estimation. Achieving remotely sensed evapotranspiration (RS_ET) with confident certainty is required but difficult. As a result, it is indispensable to develop the validation methods to quantitatively assess the accuracy and error sources of the regional RS_ET estimations. This study proposes an innovative validation method based on multi-scale evapotranspiration acquired from field measurements, with the validation results including the accuracy assessment, error source analysis, and uncertainty analysis of the validation process. It is a potentially useful approach to evaluate the accuracy and analyze the spatio-temporal properties of RS_ET at both the basin and local scales, and is appropriate to validate RS_ET in diverse resolutions at different time-scales. An independent RS_ET validation using this method was presented over the Hai River Basin, China in 2002-2009 as a case study. Validation at the basin scale showed good agreements between the 1 km annual RS_ET and the validation data such as the water balanced evapotranspiration, MODIS evapotranspiration products, precipitation, and landuse types. Validation at the local scale also had good results for monthly, daily RS_ET at 30 m and 1 km resolutions, comparing to the multi-scale evapotranspiration measurements from the EC and LAS, respectively, with the footprint model over three typical landscapes. Although some validation experiments demonstrated that the models yield accurate estimates at flux measurement sites, the question remains whether they are performing well over the broader landscape. Moreover, a large number of RS_ET products have been released in recent years. Thus, we also pay attention to the cross-validation method of RS_ET derived from multi-source models. "The Multi-scale Observation Experiment on Evapotranspiration over Heterogeneous Land Surfaces: Flux Observation Matrix" campaign is carried out at the middle reaches of the Heihe River Basin, China in 2012. Flux measurements from an observation matrix composed of 22 EC and 4 LAS are acquired to investigate the cross-validation of multi-source models over different landscapes. In this case, six remote sensing models, including the empirical statistical model, the one-source and two-source models, the Penman-Monteith equation based model, the Priestley-Taylor equation based model, and the complementary relationship based model, are used to perform an intercomparison. All the results from the two cases of RS_ET validation showed that the proposed validation methods are reasonable and feasible.

TMATS/ IHAL/ DDML Schema Validation

DTIC Science & Technology

2017-02-01

task was to create a method for performing IRIG eXtensible Markup Language (XML) schema validation. As opposed to XML instance document validation...TMATS / IHAL / DDML Schema Validation, RCC 126-17, February 2017 vii Acronyms DDML Data Display Markup Language HUD heads-up display iNET...system XML eXtensible Markup Language TMATS / IHAL / DDML Schema Validation, RCC 126-17, February 2017 viii This page intentionally left blank

Interlaboratory validation of an improved U.S. Food and Drug Administration method for detection of Cyclospora cayetanensis in produce using TaqMan real-time PCR

USDA-ARS?s Scientific Manuscript database

A collaborative validation study was performed to evaluate the performance of a new U.S. Food and Drug Administration method developed for detection of the protozoan parasite, Cyclospora cayetanensis, on cilantro and raspberries. The method includes a sample preparation step in which oocysts are re...

CFD Analysis of the SBXC Glider Airframe

DTIC Science & Technology

2016-06-01

mathematically on finite element methods. To validate and verify the methodology developed, a mathematical comparison was made with the previous research data...greater than 15 m/s. 14. SUBJECT TERMS finite element method, computational fluid dynamics, Y Plus, mesh element quality, aerodynamic data, fluid...based mathematically on finite element methods. To validate and verify the methodology developed, a mathematical comparison was made with the

Determination of vitamin C in foods: current state of method validation.

PubMed

Spínola, Vítor; Llorent-Martínez, Eulogio J; Castilho, Paula C

2014-11-21

Vitamin C is one of the most important vitamins, so reliable information about its content in foodstuffs is a concern to both consumers and quality control agencies. However, the heterogeneity of food matrixes and the potential degradation of this vitamin during its analysis create enormous challenges. This review addresses the development and validation of high-performance liquid chromatography methods for vitamin C analysis in food commodities, during the period 2000-2014. The main characteristics of vitamin C are mentioned, along with the strategies adopted by most authors during sample preparation (freezing and acidification) to avoid vitamin oxidation. After that, the advantages and handicaps of different analytical methods are discussed. Finally, the main aspects concerning method validation for vitamin C analysis are critically discussed. Parameters such as selectivity, linearity, limit of quantification, and accuracy were studied by most authors. Recovery experiments during accuracy evaluation were in general satisfactory, with usual values between 81 and 109%. However, few methods considered vitamin C stability during the analytical process, and the study of the precision was not always clear or complete. Potential future improvements regarding proper method validation are indicated to conclude this review. Copyright © 2014. Published by Elsevier B.V.

Measurement of Harm Outcomes in Older Adults after Hospital Discharge: Reliability and Validity

PubMed Central

Douglas, Alison; Letts, Lori; Eva, Kevin; Richardson, Julie

2012-01-01

Objectives. Defining and validating a measure of safety contributes to further validation of clinical measures. The objective was to define and examine the psychometric properties of the outcome “incidents of harm.” Methods. The Incident of Harm Caregiver Questionnaire was administered to caregivers of older adults discharged from hospital by telephone. Caregivers completed daily logs for one month and medical charts were examined. Results. Test-retest reliability (n = 38) was high for the occurrence of an incident of harm (yes/no; kappa = 1.0) and the type of incident (agreement = 100%). Validation against daily logs found no disagreement regarding occurrence or types of incidents. Validation with medical charts found no disagreement regarding incident occurrence and disagreement in half regarding incident type. Discussion. The data support the Incident of Harm Caregiver Questionnaire as a reliable and valid estimation of incidents for this sample and are important to researchers as a method to measure safety when validating clinical measures. PMID:22649728

What is the best method for assessing lower limb force-velocity relationship?

PubMed

Giroux, C; Rabita, G; Chollet, D; Guilhem, G

2015-02-01

This study determined the concurrent validity and reliability of force, velocity and power measurements provided by accelerometry, linear position transducer and Samozino's methods, during loaded squat jumps. 17 subjects performed squat jumps on 2 separate occasions in 7 loading conditions (0-60% of the maximal concentric load). Force, velocity and power patterns were averaged over the push-off phase using accelerometry, linear position transducer and a method based on key positions measurements during squat jump, and compared to force plate measurements. Concurrent validity analyses indicated very good agreement with the reference method (CV=6.4-14.5%). Force, velocity and power patterns comparison confirmed the agreement with slight differences for high-velocity movements. The validity of measurements was equivalent for all tested methods (r=0.87-0.98). Bland-Altman plots showed a lower agreement for velocity and power compared to force. Mean force, velocity and power were reliable for all methods (ICC=0.84-0.99), especially for Samozino's method (CV=2.7-8.6%). Our findings showed that present methods are valid and reliable in different loading conditions and permit between-session comparisons and characterization of training-induced effects. While linear position transducer and accelerometer allow for examining the whole time-course of kinetic patterns, Samozino's method benefits from a better reliability and ease of processing. © Georg Thieme Verlag KG Stuttgart · New York.

Ratiometric, single-dye, pH-sensitive inhibited laser-induced fluorescence for the characterization of mixing and mass transfer

NASA Astrophysics Data System (ADS)

Lacassagne, Tom; Simoëns, Serge; El Hajem, Mahmoud; Champagne, Jean-Yves

2018-01-01

Inhibited planar laser-induced fluorescence (I-PLIF) techniques are widely used for heat and mass transfer studies in fluid mechanics. They allow the visualization of instantaneous two-dimensional field of a passive or reactive scalar, providing that this scalar acts as an inhibitor to the fluorescence of a specific molecule, and that this molecule is homogeneously mixed in the fluid at a known concentration. Local scalar values are deduced from fluorescence recordings thanks to preliminary calibration procedure. When confronted with non-optically thin systems, however, the knowledge of the excitation intensity distribution in the region of interest is also required, and this information is most of the time hard to obtain. To overcome that problem, two-color ratiometric PLIF techniques ( {I}^ {r}-PLIF) have been developed. In these methods, the ratio of two different fluorescence wavelengths triggered by the same excitation is used as an indicator of the scalar value. Such techniques have been used for temperature measurements in several studies but never, to the author's knowledge, for pH tracking and acid-base mixing, despite the frequent use of the one-color version in mass transfer studies. In the present work, a ratiometric pH-sensitive-inhibited PLIF technique ( {I}_ {pH}^ {r}-PLIF) using fluorescein sodium as a single dye and applicable to complex geometries and flows is developed. Theoretical considerations show that the ratio of the two-color fluorescence intensities should only depend on the dye's spectral quantum yield, itself pH-dependent. A detailed spectrofluorimetric study of fluorescein reveals that this ratio strictly increases with the pH for two well-chosen spectral bands (fluorescence colors). A similar trend is found when using sCmos cameras equipped with optical filters to record fluorescence signals. The method is then experimented on a test flow, a turbulent acidic jet injected in an initially pH-neutral volume of fluid. The results obtained using the ratiometric version are consistent with single-color technique measurements, but excitation intensity heterogeneity is more efficiently accounted for, with a much smaller time needed for data treatment and without requiring the knowledge of laser paths across the fluid. This new technique is also able to reduce the impact of some unwanted experimental features such as time-varying excitation intensity or reflections at interfaces. It can be of great interest for further applications to multiphase mass transfer studies.

The use of National Weather Service Data to Compute the Dose to the MEOI.

PubMed

Vickers, Linda

2018-05-01

The Turner method is the "benchmark method" for computing the stability class that is used to compute the X/Q (s m). The Turner method should be used to ascertain the validity of X/Q results determined by other methods. This paper used site-specific meteorological data obtained from the National Weather Service. The Turner method described herein is simple, quick, accurate, and transparent because all of the data, calculations, and results are visible for verification and validation with published literature.

Validity of a Measure of Assertiveness

ERIC Educational Resources Information Center

Galassi, John P.; Galassi, Merna D.

1974-01-01

This study was concerned with further validation of a measure of assertiveness. Concurrent validity was established for the College Self-Expression Scale using the method of contrasted groups and through correlations of self-and judges' ratings of assertiveness. (Author)

Implementing an X-ray validation pipeline for the Protein Data Bank

DOE Office of Scientific and Technical Information (OSTI.GOV)

Gore, Swanand; Velankar, Sameer; Kleywegt, Gerard J., E-mail: gerard@ebi.ac.uk

2012-04-01

The implementation of a validation pipeline, based on community recommendations, for future depositions of X-ray crystal structures in the Protein Data Bank is described. There is an increasing realisation that the quality of the biomacromolecular structures deposited in the Protein Data Bank (PDB) archive needs to be assessed critically using established and powerful validation methods. The Worldwide Protein Data Bank (wwPDB) organization has convened several Validation Task Forces (VTFs) to advise on the methods and standards that should be used to validate all of the entries already in the PDB as well as all structures that will be deposited inmore » the future. The recommendations of the X-ray VTF are currently being implemented in a software pipeline. Here, ongoing work on this pipeline is briefly described as well as ways in which validation-related information could be presented to users of structural data.« less

Validity evidence based on test content.

PubMed

Sireci, Stephen; Faulkner-Bond, Molly

2014-01-01

Validity evidence based on test content is one of the five forms of validity evidence stipulated in the Standards for Educational and Psychological Testing developed by the American Educational Research Association, American Psychological Association, and National Council on Measurement in Education. In this paper, we describe the logic and theory underlying such evidence and describe traditional and modern methods for gathering and analyzing content validity data. A comprehensive review of the literature and of the aforementioned Standards is presented. For educational tests and other assessments targeting knowledge and skill possessed by examinees, validity evidence based on test content is necessary for building a validity argument to support the use of a test for a particular purpose. By following the methods described in this article, practitioners have a wide arsenal of tools available for determining how well the content of an assessment is congruent with and appropriate for the specific testing purposes.

Validation of Multilevel Constructs: Validation Methods and Empirical Findings for the EDI

ERIC Educational Resources Information Center

Forer, Barry; Zumbo, Bruno D.

2011-01-01

The purposes of this paper are to highlight the foundations of multilevel construct validation, describe two methodological approaches and associated analytic techniques, and then apply these approaches and techniques to the multilevel construct validation of a widely-used school readiness measure called the Early Development Instrument (EDI;…

Validation of Ion Chromatographic Method for Determination of Standard Inorganic Anions in Treated and Untreated Drinking Water

NASA Astrophysics Data System (ADS)

Ivanova, V.; Surleva, A.; Koleva, B.

2018-06-01

An ion chromatographic method for determination of fluoride, chloride, nitrate and sulphate in untreated and treated drinking waters was described. An automated 850 IC Professional, Metrohm system equipped with conductivity detector and Metrosep A Supp 7-250 (250 x 4 mm) column was used. The validation of the method was performed for simultaneous determination of all studied analytes and the results have showed that the validated method fits the requirements of the current water legislation. The main analytical characteristics were estimated for each of studied analytes: limits of detection, limits of quantification, working and linear ranges, repeatability and intermediate precision, recovery. The trueness of the method was estimated by analysis of certified reference material for soft drinking water. Recovery test was performed on spiked drinking water samples. An uncertainty was estimated. The method was applied for analysis of drinking waters before and after chlorination.

A review of MRI evaluation of demyelination in cuprizone murine model

NASA Astrophysics Data System (ADS)

Krutenkova, E.; Pan, E.; Khodanovich, M.

2015-11-01

The cuprizone mouse model of non-autoimmune demyelination reproduces some phenomena of multiple sclerosis and is appropriate for validation and specification of a new method of non-invasive diagnostics. In the review new data which are collected using the new MRI method are compared with one or more conventional MRI tools. Also the paper reviewed the validation of MRI approaches using histological or immunohistochemical methods. Luxol fast blue histological staining and myelin basic protein immunostaining is widespread. To improve the accuracy of non-invasive conventional MRI, multimodal scanning could be applied. The new quantitative MRI method of fast mapping of the macromolecular proton fraction is a reliable biomarker of myelin in the brain and can be used for research of demyelination in animals. To date, a validation of MPF method on the CPZ mouse model of demyelination is not performed, although this method is probably the best way to evaluate demyelination using MRI.

Approach to method development and validation in capillary electrophoresis for enantiomeric purity testing of active basic pharmaceutical ingredients.

PubMed

Sokoliess, Torsten; Köller, Gerhard

2005-06-01

A chiral capillary electrophoresis system allowing the determination of the enantiomeric purity of an investigational new drug was developed using a generic method development approach for basic analytes. The method was optimized in terms of type and concentration of both cyclodextrin (CD) and electrolyte, buffer pH, temperature, voltage, and rinsing procedure. Optimal chiral separation of the analyte was obtained using an electrolyte with 2.5% carboxymethyl-beta-CD in 25 mM NaH2PO4 (pH 4.0). Interchanging the inlet and outlet vials after each run improved the method's precision. To assure the method's suitability for the control of enantiomeric impurities in pharmaceutical quality control, its specificity, linearity, precision, accuracy, and robustness were validated according to the requirements of the International Conference on Harmonization. The usefulness of our generic method development approach for the validation of robustness was demonstrated.

International ring trial for the validation of an event-specific Golden Rice 2 quantitative real-time polymerase chain reaction method.

PubMed

Jacchia, Sara; Nardini, Elena; Bassani, Niccolò; Savini, Christian; Shim, Jung-Hyun; Trijatmiko, Kurniawan; Kreysa, Joachim; Mazzara, Marco

2015-05-27

This article describes the international validation of the quantitative real-time polymerase chain reaction (PCR) detection method for Golden Rice 2. The method consists of a taxon-specific assay amplifying a fragment of rice Phospholipase D α2 gene, and an event-specific assay designed on the 3' junction between transgenic insert and plant DNA. We validated the two assays independently, with absolute quantification, and in combination, with relative quantification, on DNA samples prepared in haploid genome equivalents. We assessed trueness, precision, efficiency, and linearity of the two assays, and the results demonstrate that both the assays independently assessed and the entire method fulfill European and international requirements for methods for genetically modified organism (GMO) testing, within the dynamic range tested. The homogeneity of the results of the collaborative trial between Europe and Asia is a good indicator of the robustness of the method.

QUANTITATIVE CANCER RISK ASSESSMENT METHODOLOGY USING SHORT-TERM GENETIC BIOASSAYS: THE COMPARATIVE POTENCY METHOD

EPA Science Inventory

Quantitative risk assessment is fraught with many uncertainties. The validity of the assumptions underlying the methods employed are often difficult to test or validate. Cancer risk assessment has generally employed either human epidemiological data from relatively high occupatio...

Validation of an isotope dilution, ICP-MS method based on internal mass bias correction for the determination of trace concentrations of Hg in sediment cores.

PubMed

Ciceri, E; Recchia, S; Dossi, C; Yang, L; Sturgeon, R E

2008-01-15

The development and validation of a method for the determination of mercury in sediments using a sector field inductively coupled plasma mass spectrometer (SF-ICP-MS) for detection is described. The utilization of isotope dilution (ID) calibration is shown to solve analytical problems related to matrix composition. Mass bias is corrected using an internal mass bias correction technique, validated against the traditional standard bracketing method. The overall analytical protocol is validated against NRCC PACS-2 marine sediment CRM. The estimated limit of detection is 12ng/g. The proposed procedure was applied to the analysis of a real sediment core sampled to a depth of 160m in Lake Como, where Hg concentrations ranged from 66 to 750ng/g.

What Do You Think You Are Measuring? A Mixed-Methods Procedure for Assessing the Content Validity of Test Items and Theory-Based Scaling

PubMed Central

Koller, Ingrid; Levenson, Michael R.; Glück, Judith

2017-01-01

The valid measurement of latent constructs is crucial for psychological research. Here, we present a mixed-methods procedure for improving the precision of construct definitions, determining the content validity of items, evaluating the representativeness of items for the target construct, generating test items, and analyzing items on a theoretical basis. To illustrate the mixed-methods content-scaling-structure (CSS) procedure, we analyze the Adult Self-Transcendence Inventory, a self-report measure of wisdom (ASTI, Levenson et al., 2005). A content-validity analysis of the ASTI items was used as the basis of psychometric analyses using multidimensional item response models (N = 1215). We found that the new procedure produced important suggestions concerning five subdimensions of the ASTI that were not identifiable using exploratory methods. The study shows that the application of the suggested procedure leads to a deeper understanding of latent constructs. It also demonstrates the advantages of theory-based item analysis. PMID:28270777

Validation of a headspace trap gas chromatography and mass spectrometry method for the quantitative analysis of volatile compounds from degraded rapeseed oil.

PubMed

Sghaier, Lilia; Cordella, Christophe B Y; Rutledge, Douglas N; Watiez, Mickaël; Breton, Sylvie; Sassiat, Patrick; Thiebaut, Didier; Vial, Jérôme

2016-05-01

Due to lipid oxidation, off-flavors, characterized by a fishy odor, are emitted during the heating of rapeseed oil in a fryer and affect the flavor of rapeseed oil even at low concentrations. Thus, there is a need for analytical methods to identify and quantify these products. To study the headspace composition of degraded rapeseed oil, and more specifically the compounds responsible for the fishy odor, a headspace trap gas chromatography with mass spectrometry method was developed and validated. Six volatile compounds formed during the degradation of rapeseed oil were quantified: 1-penten-3-one, (Z)-4-heptenal, hexanal, nonanal, (E,E)-heptadienal, and (E)-2-heptenal. Validation using accuracy profiles allowed us to determine the valid ranges of concentrations for each compound, with acceptance limits of 40% and tolerance limits of 80%. This method was then successfully applied to real samples of degraded oils. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Simultaneous quantification of five major active components in capsules of the traditional Chinese medicine ‘Shu-Jin-Zhi-Tong’ by high performance liquid chromatography

PubMed Central

Yang, Xing-Xin; Zhang, Xiao-Xia; Chang, Rui-Miao; Wang, Yan-Wei; Li, Xiao-Ni

2011-01-01

A simple and reliable high performance liquid chromatography (HPLC) method has been developed for the simultaneous quantification of five major bioactive components in ‘Shu-Jin-Zhi-Tong’ capsules (SJZTC), for the purposes of quality control of this commonly prescribed traditional Chinese medicine. Under the optimum conditions, excellent separation was achieved, and the assay was fully validated in terms of linearity, precision, repeatability, stability and accuracy. The validated method was applied successfully to the determination of the five compounds in SJZTC samples from different production batches. The HPLC method can be used as a valid analytical method to evaluate the intrinsic quality of SJZTC. PMID:29403711

Development and validation of a high performance liquid chromatography assay for 17alpha-methyltestosterone in fish feed.

PubMed

Marwah, Ashok; Marwah, Padma; Lardy, Henry

2005-09-25

17alpha-Methyltestosterone (MT) is used to manipulate the gender of a variety of fish species. A high performance liquid chromatography (HPLC) internal standard method for the determination of 17alpha-methyltestosterone in fish feed using 3beta-methoxy-17beta-hydroxyandrost-5-en-7-one as internal standard (IS) has been developed. The method has been validated for the quantitation of MT in fish feed using 245 nm UV absorbance as the parent wavelength and 255 nm as a qualifier wavelength. The method was validated in the concentration range of 15.0-120 mg/kg of 17alpha-methyltestosterone in fish feed. Method was also found to be suitable for other feeds.

Iridology: A systematic review.

PubMed

Ernst, E

1999-02-01

Iridologists claim to be able to diagnose medical conditions through abnormalities of pigmentation in the iris. This technique is popular in many countries. Therefore it is relevant to ask whether it is valid. To systematically review all interpretable tests of the validity of iridology as a diagnostic tool. DATA SOURCE AND EXTRACTION: Three independent literature searches were performed to identify all blinded tests. Data were extracted in a predefined, standardized fashion. Four case control studies were found. The majority of these investigations suggests that iridology is not a valid diagnostic method. The validity of iridology as a diagnostic tool is not supported by scientific evaluations. Patients and therapists should be discouraged from using this method.

Grid workflow validation using ontology-based tacit knowledge: A case study for quantitative remote sensing applications

NASA Astrophysics Data System (ADS)

Liu, Jia; Liu, Longli; Xue, Yong; Dong, Jing; Hu, Yingcui; Hill, Richard; Guang, Jie; Li, Chi

2017-01-01

Workflow for remote sensing quantitative retrieval is the ;bridge; between Grid services and Grid-enabled application of remote sensing quantitative retrieval. Workflow averts low-level implementation details of the Grid and hence enables users to focus on higher levels of application. The workflow for remote sensing quantitative retrieval plays an important role in remote sensing Grid and Cloud computing services, which can support the modelling, construction and implementation of large-scale complicated applications of remote sensing science. The validation of workflow is important in order to support the large-scale sophisticated scientific computation processes with enhanced performance and to minimize potential waste of time and resources. To research the semantic correctness of user-defined workflows, in this paper, we propose a workflow validation method based on tacit knowledge research in the remote sensing domain. We first discuss the remote sensing model and metadata. Through detailed analysis, we then discuss the method of extracting the domain tacit knowledge and expressing the knowledge with ontology. Additionally, we construct the domain ontology with Protégé. Through our experimental study, we verify the validity of this method in two ways, namely data source consistency error validation and parameters matching error validation.

Ultrasensitive NIR-SERRS Probes with Multiplexed Ratiometric Quantification for In Vivo Antibody Leads Validation.

PubMed

Kang, Homan; Jeong, Sinyoung; Jo, Ahla; Chang, Hyejin; Yang, Jin-Kyoung; Jeong, Cheolhwan; Kyeong, San; Lee, Youn Woo; Samanta, Animesh; Maiti, Kaustabh Kumar; Cha, Myeong Geun; Kim, Taek-Keun; Lee, Sukmook; Jun, Bong-Hyun; Chang, Young-Tae; Chung, Junho; Lee, Ho-Young; Jeong, Dae Hong; Lee, Yoon-Sik

2018-02-01

Immunotargeting ability of antibodies may show significant difference between in vitro and in vivo. To select antibody leads with high affinity and specificity, it is necessary to perform in vivo validation of antibody candidates following in vitro antibody screening. Herein, a robust in vivo validation of anti-tetraspanin-8 antibody candidates against human colon cancer using ratiometric quantification method is reported. The validation is performed on a single mouse and analyzed by multiplexed surface-enhanced Raman scattering using ultrasensitive and near infrared (NIR)-active surface-enhanced resonance Raman scattering nanoprobes (NIR-SERRS dots). The NIR-SERRS dots are composed of NIR-active labels and Au/Ag hollow-shell assembled silica nanospheres. A 93% of NIR-SERRS dots is detectable at a single-particle level and signal intensity is 100-fold stronger than that from nonresonant molecule-labeled spherical Au NPs (80 nm). The result of SERRS-based antibody validation is comparable to that of the conventional method using single-photon-emission computed tomography. The NIR-SERRS-based strategy is an alternate validation method which provides cost-effective and accurate multiplexing measurements for antibody-based drug development. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Update of Standard Practices for New Method Validation in Forensic Toxicology.

PubMed

Wille, Sarah M R; Coucke, Wim; De Baere, Thierry; Peters, Frank T

2017-01-01

International agreement concerning validation guidelines is important to obtain quality forensic bioanalytical research and routine applications as it all starts with the reporting of reliable analytical data. Standards for fundamental validation parameters are provided in guidelines as those from the US Food and Drug Administration (FDA), the European Medicines Agency (EMA), the German speaking Gesellschaft fur Toxikologie und Forensische Chemie (GTFCH) and the Scientific Working Group of Forensic Toxicology (SWGTOX). These validation parameters include selectivity, matrix effects, method limits, calibration, accuracy and stability, as well as other parameters such as carryover, dilution integrity and incurred sample reanalysis. It is, however, not easy for laboratories to implement these guidelines into practice as these international guidelines remain nonbinding protocols, that depend on the applied analytical technique, and that need to be updated according the analyst's method requirements and the application type. In this manuscript, a review of the current guidelines and literature concerning bioanalytical validation parameters in a forensic context is given and discussed. In addition, suggestions for the experimental set-up, the pros and cons of statistical approaches and adequate acceptance criteria for the validation of bioanalytical applications are given. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

Measuring adverse events in helicopter emergency medical services: establishing content validity.

PubMed

Patterson, P Daniel; Lave, Judith R; Martin-Gill, Christian; Weaver, Matthew D; Wadas, Richard J; Arnold, Robert M; Roth, Ronald N; Mosesso, Vincent N; Guyette, Francis X; Rittenberger, Jon C; Yealy, Donald M

2014-01-01

We sought to create a valid framework for detecting adverse events (AEs) in the high-risk setting of helicopter emergency medical services (HEMS). We assembled a panel of 10 expert clinicians (n = 6 emergency medicine physicians and n = 4 prehospital nurses and flight paramedics) affiliated with a large multistate HEMS organization in the Northeast US. We used a modified Delphi technique to develop a framework for detecting AEs associated with the treatment of critically ill or injured patients. We used a widely applied measure, the content validity index (CVI), to quantify the validity of the framework's content. The expert panel of 10 clinicians reached consensus on a common AE definition and four-step protocol/process for AE detection in HEMS. The consensus-based framework is composed of three main components: (1) a trigger tool, (2) a method for rating proximal cause, and (3) a method for rating AE severity. The CVI findings isolate components of the framework considered content valid. We demonstrate a standardized process for the development of a content-valid framework for AE detection. The framework is a model for the development of a method for AE identification in other settings, including ground-based EMS.

A validation method for near-infrared spectroscopy based tissue oximeters for cerebral and somatic tissue oxygen saturation measurements.

PubMed

Benni, Paul B; MacLeod, David; Ikeda, Keita; Lin, Hung-Mo

2018-04-01

We describe the validation methodology for the NIRS based FORE-SIGHT ELITE ® (CAS Medical Systems, Inc., Branford, CT, USA) tissue oximeter for cerebral and somatic tissue oxygen saturation (StO 2 ) measurements for adult subjects submitted to the United States Food and Drug Administration (FDA) to obtain clearance for clinical use. This validation methodology evolved from a history of NIRS validations in the literature and FDA recommended use of Deming regression and bootstrapping statistical validation methods. For cerebral validation, forehead cerebral StO 2 measurements were compared to a weighted 70:30 reference (REF CX B ) of co-oximeter internal jugular venous and arterial blood saturation of healthy adult subjects during a controlled hypoxia sequence, with a sensor placed on the forehead. For somatic validation, somatic StO 2 measurements were compared to a weighted 70:30 reference (REF CX S ) of co-oximetry central venous and arterial saturation values following a similar protocol, with sensors place on the flank, quadriceps muscle, and calf muscle. With informed consent, 25 subjects successfully completed the cerebral validation study. The bias and precision (1 SD) of cerebral StO 2 compared to REF CX B was -0.14 ± 3.07%. With informed consent, 24 subjects successfully completed the somatic validation study. The bias and precision of somatic StO 2 compared to REF CX S was 0.04 ± 4.22% from the average of flank, quadriceps, and calf StO 2 measurements to best represent the global whole body REF CX S . The NIRS validation methods presented potentially provide a reliable means to test NIRS monitors and qualify them for clinical use.

In-house validation of a liquid chromatography-tandem mass spectrometry method for the determination of selective androgen receptor modulators (SARMS) in bovine urine.

PubMed

Schmidt, Kathrin S; Mankertz, Joachim

2018-06-01

A sensitive and robust LC-MS/MS method allowing the rapid screening and confirmation of selective androgen receptor modulators in bovine urine was developed and successfully validated according to Commission Decision 2002/657/EC, chapter 3.1.3 'alternative validation', by applying a matrix-comprehensive in-house validation concept. The confirmation of the analytes in the validation samples was achieved both on the basis of the MRM ion ratios as laid down in Commission Decision 2002/657/EC and by comparison of their enhanced product ion (EPI) spectra with a reference mass spectral library by making use of the QTRAP technology. Here, in addition to the MRM survey scan, EPI spectra were generated in a data-dependent way according to an information-dependent acquisition criterion. Moreover, stability studies of the analytes in solution and in matrix according to an isochronous approach proved the stability of the analytes in solution and in matrix for at least the duration of the validation study. To identify factors that have a significant influence on the test method in routine analysis, a factorial effect analysis was performed. To this end, factors considered to be relevant for the method in routine analysis (e.g. operator, storage duration of the extracts before measurement, different cartridge lots and different hydrolysis conditions) were systematically varied on two levels. The examination of the extent to which these factors influence the measurement results of the individual analytes showed that none of the validation factors exerts a significant influence on the measurement results.

Validation of Gujarati Version of ABILOCO-Kids Questionnaire

PubMed Central

Diwan, Jasmin; Patel, Pankaj; Bansal, Ankita B.

2015-01-01

Background ABILOCO-Kids is a measure of locomotion ability for children with cerebral palsy (CP) aged 6 to 15 years & is available in English & French. Aim To validate the Gujarati version of ABILOCO-Kids questionnaire to be used in clinical research on Gujarati population. Materials and Methods ABILOCO-Kids questionnaire was translated into Gujarati from English using forward-backward-forward method. To ensure face & content validity of Gujarati version using group consensus method, each item was examined by group of experts having mean experience of 24.62 years in field of paediatric and paediatric physiotherapy. Each item was analysed for content, meaning, wording, format, ease of administration & scoring. Each item was scored by expert group as either accepted, rejected or accepted with modification. Procedure was continued until 80% of consensus for all items. Concurrent validity was examined on 55 children with Cerebral Palsy (6-15 years) of all Gross Motor Functional Classification System (GMFCS) level & all clinical types by correlating score of ABILOCO-Kids with Gross Motor Functional Measure & GMFCS. Result In phase 1 of validation, 16 items were accepted as it is; 22 items accepted with modification & 3 items went for phase 2 validation. For concurrent validity, highly significant positive correlation was found between score of ABILOCO-Kids & total GMFM (r=0.713, p<0.005) & highly significant negative correlation with GMFCS (r= -0.778, p<0.005). Conclusion Gujarati translated version of ABILOCO-Kids questionnaire has good face & content validity as well as concurrent validity which can be used to measure caregiver reported locomotion ability in children with CP. PMID:26557603

Assessment of the Validity of the Research Diagnostic Criteria for Temporomandibular Disorders: Overview and Methodology

PubMed Central

Schiffman, Eric L.; Truelove, Edmond L.; Ohrbach, Richard; Anderson, Gary C.; John, Mike T.; List, Thomas; Look, John O.

2011-01-01

AIMS The purpose of the Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD) Validation Project was to assess the diagnostic validity of this examination protocol. An overview is presented, including Axis I and II methodology and descriptive statistics for the study participant sample. This paper details the development of reliable methods to establish the reference standards for assessing criterion validity of the Axis I RDC/TMD diagnoses. Validity testing for the Axis II biobehavioral instruments was based on previously validated reference standards. METHODS The Axis I reference standards were based on the consensus of 2 criterion examiners independently performing a comprehensive history, clinical examination, and evaluation of imaging. Intersite reliability was assessed annually for criterion examiners and radiologists. Criterion exam reliability was also assessed within study sites. RESULTS Study participant demographics were comparable to those of participants in previous studies using the RDC/TMD. Diagnostic agreement of the criterion examiners with each other and with the consensus-based reference standards was excellent with all kappas ≥ 0.81, except for osteoarthrosis (moderate agreement, k = 0.53). Intrasite criterion exam agreement with reference standards was excellent (k ≥ 0.95). Intersite reliability of the radiologists for detecting computed tomography-disclosed osteoarthrosis and magnetic resonance imaging-disclosed disc displacement was good to excellent (k = 0.71 and 0.84, respectively). CONCLUSION The Validation Project study population was appropriate for assessing the reliability and validity of the RDC/TMD Axis I and II. The reference standards used to assess the validity of Axis I TMD were based on reliable and clinically credible methods. PMID:20213028

Development of a framework for international certification by OIE of diagnostic tests validated as fit for purpose.

PubMed

Wright, P; Edwards, S; Diallo, A; Jacobson, R

2006-01-01

Historically, the OIE has focused on test methods applicable to trade and the international movement of animals and animal products. With its expanding role as the World Organisation for Animal Health, the OIE has recognised the need to evaluate test methods relative to specific diagnostic applications other than trade. In collaboration with its international partners, the OIE solicited input from experts through consultants' meetings on the development of guidelines for validation and certification of diagnostic assays for infectious animal diseases. Recommendations from the first meeting were formally adopted and have subsequently been acted upon by the OIE. A validation template has been developed that specifically requires a test to be fit or suited for its intended purpose (e.g. as a screening or a confirmatory test). This is a key criterion for validation. The template incorporates four distinct stages of validation, each of which has bearing on the evaluation of fitness for purpose. The OIE has just recently created a registry for diagnostic tests that fulfil these validation requirements. Assay developers are invited to submit validation dossiers to the OIE for evaluation by a panel of experts. Recognising that validation is an incremental process, tests methods achieving at least the first stages of validation may be provisionally accepted. To provide additional confidence in assay performance, the OIE, through its network of Reference Laboratories, has embarked on the development of evaluation panels. These panels would contain specially selected test samples that would assist in verifying fitness for purpose.

Development of a framework for international certification by the OIE of diagnostic tests validated as fit for purpose.

PubMed

Wright, P; Edwards, S; Diallo, A; Jacobson, R

2007-01-01

Historically, the OIE has focussed on test methods applicable to trade and the international movement of animals and animal products. With its expanding role as the World Organisation for Animal Health, the OIE has recognised the need to evaluate test methods relative to specific diagnostic applications other than trade. In collaboration with its international partners, the OIE solicited input from experts through consultants meetings on the development of guidelines for validation and certification of diagnostic assays for infectious animal diseases. Recommendations from the first meeting were formally adopted and have subsequently been acted upon by the OIE. A validation template has been developed that specifically requires a test to be fit or suited for its intended purpose (e.g. as a screening or a confirmatory test). This is a key criterion for validation. The template incorporates four distinct stages of validation, each of which has bearing on the evaluation of fitness for purpose. The OIE has just recently created a registry for diagnostic tests that fulfil these validation requirements. Assay developers are invited to submit validation dossiers to the OIE for evaluation by a panel of experts. Recognising that validation is an incremental process, tests methods achieving at least the first stages of validation may be provisionally accepted. To provide additional confidence in assay performance, the OIE, through its network of Reference Laboratories, has embarked on the development of evaluation panels. These panels would contain specially selected test samples that would assist in verifying fitness for purpose.

Validity and reliability of the Myotest accelerometric system for the assessment of vertical jump height.

PubMed

Casartelli, Nicola; Müller, Roland; Maffiuletti, Nicola A

2010-11-01

The aim of the present study was to verify the validity and reliability of the Myotest accelerometric system (Myotest SA, Sion, Switzerland) for the assessment of vertical jump height. Forty-four male basketball players (age range: 9-25 years) performed series of squat, countermovement and repeated jumps during 2 identical test sessions separated by 2-15 days. Flight height was simultaneously quantified with the Myotest system and validated photoelectric cells (Optojump). Two calculation methods were used to estimate the jump height from Myotest recordings: flight time (Myotest-T) and vertical takeoff velocity (Myotest-V). Concurrent validity was investigated comparing Myotest-T and Myotest-V to the criterion method (Optojump), and test-retest reliability was also examined. As regards validity, Myotest-T overestimated jumping height compared to Optojump (p < 0.001) with a systematic bias of approximately 7 cm, even though random errors were low (2.7 cm) and intraclass correlation coefficients (ICCs) where high (>0.98), that is, excellent validity. Myotest-V overestimated jumping height compared to Optojump (p < 0.001), with high random errors (>12 cm), high limits of agreement ratios (>36%), and low ICCs (<0.75), that is, poor validity. As regards reliability, Myotest-T showed high ICCs (range: 0.92-0.96), whereas Myotest-V showed low ICCs (range: 0.56-0.89), and high random errors (>9 cm). In conclusion, Myotest-T is a valid and reliable method for the assessment of vertical jump height, and its use is legitimate for field-based evaluations, whereas Myotest-V is neither valid nor reliable.

Learning to recognize rat social behavior: Novel dataset and cross-dataset application.

PubMed

Lorbach, Malte; Kyriakou, Elisavet I; Poppe, Ronald; van Dam, Elsbeth A; Noldus, Lucas P J J; Veltkamp, Remco C

2018-04-15

Social behavior is an important aspect of rodent models. Automated measuring tools that make use of video analysis and machine learning are an increasingly attractive alternative to manual annotation. Because machine learning-based methods need to be trained, it is important that they are validated using data from different experiment settings. To develop and validate automated measuring tools, there is a need for annotated rodent interaction datasets. Currently, the availability of such datasets is limited to two mouse datasets. We introduce the first, publicly available rat social interaction dataset, RatSI. We demonstrate the practical value of the novel dataset by using it as the training set for a rat interaction recognition method. We show that behavior variations induced by the experiment setting can lead to reduced performance, which illustrates the importance of cross-dataset validation. Consequently, we add a simple adaptation step to our method and improve the recognition performance. Most existing methods are trained and evaluated in one experimental setting, which limits the predictive power of the evaluation to that particular setting. We demonstrate that cross-dataset experiments provide more insight in the performance of classifiers. With our novel, public dataset we encourage the development and validation of automated recognition methods. We are convinced that cross-dataset validation enhances our understanding of rodent interactions and facilitates the development of more sophisticated recognition methods. Combining them with adaptation techniques may enable us to apply automated recognition methods to a variety of animals and experiment settings. Copyright © 2017 Elsevier B.V. All rights reserved.

Quantitative determination and sampling of azathioprine residues for cleaning validation in production area.

PubMed

Fazio, Tatiana Tatit; Singh, Anil Kumar; Kedor-Hackmann, Erika Rosa Maria; Santoro, Maria Inês Rocha Miritello

2007-03-12

Cleaning validation is an integral part of current good manufacturing practices in any pharmaceutical industry. Nowadays, azathioprine and several other pharmacologically potent pharmaceuticals are manufactured in same production area. Carefully designed cleaning validation and its evaluation can ensure that residues of azathioprine will not carry over and cross contaminate the subsequent product. The aim of this study was to validate simple analytical method for verification of residual azathioprine in equipments used in the production area and to confirm efficiency of cleaning procedure. The HPLC method was validated on a LC system using Nova-Pak C18 (3.9 mm x 150 mm, 4 microm) and methanol-water-acetic acid (20:80:1, v/v/v) as mobile phase at a flow rate of 1.0 mL min(-1). UV detection was made at 280 nm. The calibration curve was linear over a concentration range from 2.0 to 22.0 microg mL(-1) with a correlation coefficient of 0.9998. The detection limit (DL) and quantitation limit (QL) were 0.09 and 0.29 microg mL(-1), respectively. The intra-day and inter-day precision expressed as relative standard deviation (R.S.D.) were below 2.0%. The mean recovery of method was 99.19%. The mean extraction-recovery from manufacturing equipments was 83.5%. The developed UV spectrophotometric method could only be used as limit method to qualify or reject cleaning procedure in production area. Nevertheless, the simplicity of spectrophotometric method makes it useful for routine analysis of azathioprine residues on cleaned surface and as an alternative to proposed HPLC method.

Inter-Disciplinary Collaboration in Support of the Post-Standby TREAT Mission

DOE Office of Scientific and Technical Information (OSTI.GOV)

DeHart, Mark; Baker, Benjamin; Ortensi, Javier

Although analysis methods have advanced significantly in the last two decades, high fidelity multi- physics methods for reactors systems have been under development for only a few years and are not presently mature nor deployed. Furthermore, very few methods provide the ability to simulate rapid transients in three dimensions. Data for validation of advanced time-dependent multi- physics is sparse; at TREAT, historical data were not collected for the purpose of validating three-dimensional methods, let alone multi-physics simulations. Existing data continues to be collected to attempt to simulate the behavior of experiments and calibration transients, but it will be insufficient formore » the complete validation of analysis methods used for TREAT transient simulations. Hence, a 2018 restart will most likely occur without the direct application of advanced modeling and simulation methods. At present, the current INL modeling and simulation team plans to work with TREAT operations staff in performing reactor simulations with MAMMOTH, in parallel with the software packages currently being used in preparation for core restart (e.g., MCNP5, RELAP5, ABAQUS). The TREAT team has also requested specific measurements to be performed during startup testing, currently scheduled to run from February to August of 2018. These startup measurements will be crucial in validating the new analysis methods in preparation for ultimate application for TREAT operations and experiment design. This document describes the collaboration between modeling and simulation staff and restart, operations, instrumentation and experiment development teams to be able to effectively interact and achieve successful validation work during restart testing.« less