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Sample records for acid titration method

  1. A new method for determining the acid number of biodiesel based on coulometric titration.

    PubMed

    Barbieri Gonzaga, Fabiano; Pereira Sobral, Sidney

    2012-08-15

    A new method is proposed for determining the acid number (AN) of biodiesel using coulometric titration with potentiometric detection, basically employing a potentiostat/galvanostat and an electrochemical cell containing a platinum electrode, a silver electrode, and a combination pH electrode. The method involves a sequential application of a constant current between the platinum (cathode) and silver (anode) electrodes, followed by measuring the potential of the combination pH electrode, using an isopropanol/water mixture as solvent and LiCl as the supporting electrolyte. A preliminary evaluation of the new method, using acetic acid for doping a biodiesel sample, showed an average recovery of 100.1%. Compared to a volumetric titration-based method for determining the AN of several biodiesel samples (ranging from about 0.18 to 0.95 mg g(-1)), the new method produced statistically similar results with better repeatability. Compared to other works reported in the literature, the new method presented an average repeatability up to 3.2 times better and employed a sample size up to 20 times smaller.

  2. Radiometric acid-base titrations.

    PubMed

    Erdey, L; Gimesi, O; Szabadváry, F

    1969-03-01

    Acid-base titrations can be performed with radiometric end-point detection by use of labelled metal salts (e.g., ZnCl(2), HgCl(2)). Owing to the formation or dissolution of the corresponding hydroxide after the equivalence point, the activity of the titrated solution linearly increases or decreases as excess of standard solution is added. The end-point of the titration is determined graphically.

  3. Acid-base titrations by stepwise addition of equal volumes of titrant with special reference to automatic titrations-IV Photometric titration of an acid.

    PubMed

    Pehrsson, L; Ingman, F

    1977-02-01

    In Part I of this series, evaluation methods for potentiometric titrations of acids performed by stepwise addition of titrant were introduced. These methods are non-approximative and may therefore in principle be applied to titrations of acids of any strength. This paper demonstrates that photometric titrations performed by stepwise addition of titrant may also be evaluated by the proposed methods. The photometric method is compared with the potentiometric method of monitoring the course of a titration.

  4. Theoretical considerations and a simple method for measuring alkalinity and acidity in low-pH waters by gran titration

    USGS Publications Warehouse

    Barringer, J.L.; Johnsson, P.A.

    1996-01-01

    Titrations for alkalinity and acidity using the technique described by Gran (1952, Determination of the equivalence point in potentiometric titrations, Part II: The Analyst, v. 77, p. 661-671) have been employed in the analysis of low-pH natural waters. This report includes a synopsis of the theory and calculations associated with Gran's technique and presents a simple and inexpensive method for performing alkalinity and acidity determinations. However, potential sources of error introduced by the chemical character of some waters may limit the utility of Gran's technique. Therefore, the cost- and time-efficient method for performing alkalinity and acidity determinations described in this report is useful for exploring the suitability of Gran's technique in studies of water chemistry.

  5. Acid-base titrations by stepwise additions of equal volumes of titrant with special reference to automatic titrations-I Theory, discussion of the gran functions, the hofstee method and two proposed methods for calculating equivalence volumes.

    PubMed

    Pehrsson, L; Ingman, F; Johansson, A

    The range of acid stability constants over which the Gran functions are applicable in evaluating acid-base titrations at normal concentrations is studied and found to be quite narrow. Alternatively, the titrations can be evaluated by using non-approximative methods based upon an equation that is derived without making the simplifying assumptions leading to the Gran functions. One such method, introduced by Hofstee, is discussed and the results of a study of the effects of a systematic error in the pH-determinations upon the determination of the equivalence volume are given. Two other non-approximative methods for evaluating acid-base titrations that are particularly suitable for use in automatic titrations of many similar samples are also described. These methods yield correct values of the equivalence volume regardless of a possible systematic error in the pH-values and are particularly useful for titrations of moderately strong and very weak acids (stability constants up to 10(10)-10(11) at 0.01M concentration) where the Gran functions cannot be used.

  6. Acid Rain Analysis by Standard Addition Titration.

    ERIC Educational Resources Information Center

    Ophardt, Charles E.

    1985-01-01

    The standard addition titration is a precise and rapid method for the determination of the acidity in rain or snow samples. The method requires use of a standard buret, a pH meter, and Gran's plot to determine the equivalence point. Experimental procedures used and typical results obtained are presented. (JN)

  7. A digital image-based method for determining of total acidity in red wines using acid-base titration without indicator.

    PubMed

    Tôrres, Adamastor Rodrigues; Lyra, Wellington da Silva; de Andrade, Stéfani Iury Evangelista; Andrade, Renato Allan Navarro; da Silva, Edvan Cirino; Araújo, Mário César Ugulino; Gaião, Edvaldo da Nóbrega

    2011-05-15

    This work proposes the use of digital image-based method for determination of total acidity in red wines by means of acid-base titration without using an external indicator or any pre-treatment of the sample. Digital images present the colour of the emergent radiation which is complementary to the radiation absorbed by anthocyanines present in wines. Anthocyanines change colour depending on the pH of the medium, and from the variation of colour in the images obtained during titration, the end point can be localized with accuracy and precision. RGB-based values were employed to build titration curves, and end points were localized by second derivative curves. The official method recommends potentiometric titration with a NaOH standard solution, and sample dilution until the pH reaches 8.2-8.4. In order to illustrate the feasibility of the proposed method, titrations of ten red wines were carried out. Results were compared with the reference method, and no statistically significant difference was observed between the results by applying the paired t-test at the 95% confidence level. The proposed method yielded more precise results than the official method. This is due to the trivariate nature of the measurements (RGB), associated with digital images.

  8. CONDUCTIVITY TITRATION OF GELATIN SOLUTIONS WITH ACIDS.

    PubMed

    Hitchcock, D I

    1923-11-20

    Titrations have been made, by the conductivity method, of gelatin solutions with hydrochloric and sulphuric acids. The results indicate an end-point at about 8.6 cc. of N/10 acid per gm. of gelatin, or a combining weight of about 1,160. These results are in fair agreement with those previously obtained by the hydrogen electrode method. Better agreement between the two methods was found in the case of deaminized gelatin. The data are in accord with a purely chemical conception of the combination between protein and acid.

  9. Development of High-purity Certified Reference Materials for 17 Proteinogenic Amino Acids by Traceable Titration Methods.

    PubMed

    Kato, Megumi; Yamazaki, Taichi; Kato, Hisashi; Eyama, Sakae; Goto, Mari; Yoshioka, Mariko; Takatsu, Akiko

    2015-01-01

    To ensure the reliability of amino acid analyses, the National Metrology Institute of Japan of the National Institute of Advanced Industrial Science and Technology (NMIJ/AIST) has developed high-purity certified reference materials (CRMs) for 17 proteinogenic amino acids. These CRMs are intended for use as primary reference materials to enable the traceable quantification of amino acids. The purity of the present CRMs was determined based on two traceable methods: nonaqueous acidimetric titration and nitrogen determination by the Kjeldahl method. Since neither method could distinguish compounds with similar structures, such as amino acid-related impurities, impurities were thoroughly quantified by combining several HPLC methods, and subtracted from the obtained purity of each method. The property value of each amino acid was calculated as a weighted mean of the corrected purities by the two methods. The uncertainty of the property value was obtained by combining measurement uncertainties of the two methods, a difference between the two methods, the uncertainty from the contribution of impurities, and the uncertainty derived from inhomogeneity. The uncertainty derived from instability was considered to be negligible based on stability monitoring of some CRMs. The certified value of each amino acid, property value with uncertainty, was given for both with or without enantiomeric separation.

  10. Sequential injection titration method using second-order signals: determination of acidity in plant oils and biodiesel samples.

    PubMed

    del Río, Vanessa; Larrechi, M Soledad; Callao, M Pilar

    2010-06-15

    A new concept of flow titration is proposed and demonstrated for the determination of total acidity in plant oils and biodiesel. We use sequential injection analysis (SIA) with a diode array spectrophotometric detector linked to chemometric tools such as multivariate curve resolution-alternating least squares (MCR-ALS). This system is based on the evolution of the basic specie of an acid-base indicator, alizarine, when it comes into contact with a sample that contains free fatty acids. The gradual pH change in the reactor coil due to diffusion and reaction phenomenona allows the sequential appearance of both species of the indicator in the detector coil, recording a data matrix for each sample. The SIA-MCR-ALS method helps to reduce the amounts of sample, the reagents and the time consumed. Each determination consumes 0.413ml of sample, 0.250ml of indicator and 3ml of carrier (ethanol) and generates 3.333ml of waste. The frequency of the analysis is high (12 samples h(-1) including all steps, i.e., cleaning, preparing and analysing). The utilized reagents are of common use in the laboratory and it is not necessary to use the reagents of perfect known concentration. The method was applied to determine acidity in plant oil and biodiesel samples. Results obtained by the proposed method compare well with those obtained by the official European Community method that is time consuming and uses large amounts of organic solvents.

  11. Comparison of a miniaturized shake-flask solubility method with automated potentiometric acid/base titrations and calculated solubilities.

    PubMed

    Glomme, A; März, J; Dressman, J B

    2005-01-01

    Solubility is one of the most important parameters for lead selection and optimization during drug discovery. Its determination should therefore take place as early as possible in the process. Because of the large numbers of compounds involved and the very low amounts of each compound available in the early development stage, it is highly desirable to measure the solubility with as little compound as possible and to be able to improve the throughput of the methods used. In this work, a miniaturized shake-flask method was developed and the solubility results were compared with those measured by semiautomated potentiometric acid/base titrations and computational methods for 21 poorly soluble compounds with solubilities mostly in the range 0.03-30 microg/mL. The potentiometric method is very economical (approximately 100 microg of a poorly soluble compound is needed) and is able to create a pH/solubility profile with one single determination, but is limited to ionizable compounds. The miniaturized shake-flask method can be used for all compounds and a wide variety of media. Its precision and throughput proved superior to the potentiometric method for very poorly soluble compounds. Up to 20 compounds a week can be studied with one set-up. Calculated solubility data seem to be sufficient for a first estimate of the solubility, but they cannot currently be used as a substitute for experimental measurements at key decision points in the development process.

  12. The acid-base titration of montmorillonite

    NASA Astrophysics Data System (ADS)

    Bourg, I. C.; Sposito, G.; Bourg, A. C.

    2003-12-01

    Proton binding to clay minerals plays an important role in the chemical reactivity of soils (e.g., acidification, retention of nutrients or pollutants). If should also affect the performance of clay barriers for waste disposal. The surface acidity of clay minerals is commonly modelled empirically by assuming generic amphoteric surface sites (>SOH) on a flat surface, with fitted site densities and acidity constant. Current advances in experimental methods (notably spectroscopy) are rapidly improving our understanding of the structure and reactivity of the surface of clay minerals (arrangement of the particles, nature of the reactive surface sites, adsorption mechanisms). These developments are motivated by the difficulty of modelling the surface chemistry of mineral surfaces at the macro-scale (e.g., adsorption or titration) without a detailed (molecular-scale) picture of the mechanisms, and should be progressively incorporated into surface complexation models. In this view, we have combined recent estimates of montmorillonite surface properties (surface site density and structure, edge surface area, surface electrostatic potential) with surface site acidities obtained from the titration of alpha-Al2O3 and SiO2, and a novel method of accounting for the unknown initial net proton surface charge of the solid. The model predictions were compared to experimental titrations of SWy-1 montmorillonite and purified MX-80 bentonite in 0.1-0.5 mol/L NaClO4 and 0.005-0.5 mol/L NaNO3 background electrolytes, respectively. Most of the experimental data were appropriately described by the model after we adjusted a single parameter (silanol sites on the surface of montmorillonite were made to be slightly more acidic than those of silica). At low ionic strength and acidic pH the model underestimated the buffering capacity of the montmorillonite, perhaps due to clay swelling or to the interlayer adsorption of dissolved aluminum. The agreement between our model and the experimental

  13. Comparison of the serial dilution indicator and intragastric titration methods for measurement of meal-stimulated gastric acid secretion in man.

    PubMed

    Hogan, D L; Turken, D; Stern, A I; Isenberg, J I

    1983-11-01

    Two in vivo methods that permit quantitation of gastric acid secretion immediately after the meal are currently in use: intragastric titration and the serial dilution indicator method. During intragastric titration, intragastric pH is artificially maintained at 5.5 to 7 by the continuous addition of alkali to the gastric contents, while during serial dilution the intragastric pH is permitted to seek its natural pH. This study compared gastric acid secretion and serum gastrin in response to a liquid protein meal measured by both techniques in 10 subjects. Mean (+/- SE) 3-hr acid outputs were almost identical (53.6 +/- 6.0 mmol/3 hr with intragastric titration and 52.0 +/- 8.5 mmol/3 hr with serial dilution indicator). Furthermore, 30 min secretory responses in individual subjects were highly correlated (r = 0.98 +/- 0.01, P less than 0.001). Also, in spite of intragastric pH being less than 1.5 by 90 min after the meal during the serial dilution method, total integrated serum gastrin concentrations after the meal were similar (intragastric titration = 20.6 +/- 7.3 ng min/ml versus serial dilution indicator = 23.5 +/- 9.8 ng min/ml) and individual 30-min gastrins during the two separate tests were highly correlated (r = 0.80 +/- 0.06, P less than 0.01). It is concluded that both meal-stimulated gastric acid secretion and serum gastrin concentrations as measured by intragastric titration and by the serial dilution indicator method produced similar results.

  14. Logarithmic diagrams in acid-base titrations and estimation of titration errors.

    PubMed

    Wänninen, E

    1980-01-01

    The use of a logarithmic diagram for the estimation of the pH-value at the equivalence point and the titration error when a solution containing one or two acids is titrated with standard alkali is described.

  15. A comparison of computer methods for seawater alkalinity titrations

    NASA Astrophysics Data System (ADS)

    Barron, J. L.; Dyrssen, D.; Jones, E. P.; Wedborg, M.

    1983-04-01

    Potentiometric hydrochloric acid titration of seawater provides a powerful technique for determining components of the carbonate system. Recently, questions have been raised regarding older computer procedures for extracting the carbonate system parameters from the titration curve. We compare four evaluation methods, an early Gran method, the GEOSECS Gran method, a new modified Gran method, and a curve-fitting method. We conclude that the new modified Gran method and the curve-fitting can result in a precision of better than 0.1% but because of possible problems associated with representing all relevant chemical reactions during titration, an alkalinity standard must be established before accuracies of 0.1% can be achieved.

  16. Determination of Acidity Constants by Gradient Flow-Injection Titration

    ERIC Educational Resources Information Center

    Conceicao, Antonio C. L.; Minas da Piedade, Manuel E.

    2006-01-01

    A three-hour laboratory experiment, designed for an advanced undergraduate course in instrumental analysis that illustrates the application of the gradient chamber flow-injection titration (GCFIT) method with spectrophotometric detection to determine acidity constants is presented. The procedure involves the use of an acid-base indicator to obtain…

  17. Novel titration method for surface-functionalised silica

    NASA Astrophysics Data System (ADS)

    Hofen, Kai; Weber, Siegfried; Chan, Chiu Ping Candace; Majewski, Peter

    2011-01-01

    This paper describes three inexpensive and fast analytical methods to characterise grafted particle surfaces. The reaction of silica with (3-aminopropyl)triethoxysilane, (3-mercaptopropyl)trimethoxysilane and N-(phosphonomethyl)iminodiacetic acid hydrate, respectively, leads to NH2-, SO3H- or COOH-functionalised silica, which were characterised by X-ray photoelectron spectrometry and titration in nonaqueous media as well as with two titration methods in a water-based environment. In the work presented, factors influencing the titrations are pointed out and solutions are presented to overcome these limiting factors are shown.

  18. An Olfactory Indicator for Acid-Base Titrations.

    ERIC Educational Resources Information Center

    Flair, Mark N.; Setzer, William N.

    1990-01-01

    The use of an olfactory acid-base indicator in titrations for visually impaired students is discussed. Potential olfactory indicators include eugenol, thymol, vanillin, and thiophenol. Titrations performed with each indicator with eugenol proved to be successful. (KR)

  19. The Acid-Base Titration of a Very Weak Acid: Boric Acid

    ERIC Educational Resources Information Center

    Celeste, M.; Azevedo, C.; Cavaleiro, Ana M. V.

    2012-01-01

    A laboratory experiment based on the titration of boric acid with strong base in the presence of d-mannitol is described. Boric acid is a very weak acid and direct titration with NaOH is not possible. An auxiliary reagent that contributes to the release of protons in a known stoichiometry facilitates the acid-base titration. Students obtain the…

  20. Simple and Automated Coulometric Titration of Acid Using Nonisolated Electrodes

    ERIC Educational Resources Information Center

    Kuntzleman, Thomas S.; Kenney, Joshua B.; Hasbrouck, Scott; Collins, Michael J.; Amend, John R.

    2011-01-01

    Coulometric titrations involve the quantification of analyte by measurements of current and time. In most coulometric titrations, the anode and cathode are placed in isolated cells that are connected by a salt bridge. By contrast, the experiments described here involve coulometric titrations (of acidic protons in solution) using a silver anode and…

  1. Computer-assisted Gran titration procedure for strong acid determination

    SciTech Connect

    Phillips, M.F.; Gaffney, J.S.; Goodrich, R.W.; Tanner, R.L.

    1984-10-01

    An automated method for determining, by coulometric titration, small amounts of strong acid in the presence of weak acids is given. Essentially, a pH meter and a coulometer are coupled with a Tektronix 4052 mini-computer, and a two-step computer program then directs the titration and calculates the equivalence point by the method of Gran. A comparison of precision and accuracy of results for test solutions by manual and automated data reduction methods is presented. The method is being used successfully to analyze for the H/sup +/ content in ambient aerosol samples from aerometric field experiments, and can be used for cloud and rainwater samples as well. 3 references, 1 figure, 1 table.

  2. Acid-base titrations by stepwise addition of equal volumes of titrant with special reference to automatic titrations-II Theory of titration of mixtures of acids, polyprotic acids, acids in mixture with weak bases, and ampholytes.

    PubMed

    Pehrsson, L; Ingman, F; Johansson, S

    A general method for evaluating titration data for mixtures of acids and for acids in mixture with weak bases is presented. Procedures are given that do not require absolute [H]-data, i.e., relative [H]-data may be used. In most cases a very rough calibration of the electrode system is enough. Further, for simple systems, very approximate values of the stability constants are sufficient. As examples, the titration of the following are treated in some detail: a mixture of two acids, a diprotic acid, an acid in presence of its conjugate base, and an ampholyte.

  3. Acid Base Titrations in Nonaqueous Solvents and Solvent Mixtures

    NASA Astrophysics Data System (ADS)

    Barcza, Lajos; Buvári-Barcza, Ágnes

    2003-07-01

    The acid base determination of different substances by nonaqueous titrations is highly preferred in pharmaceutical analyses since the method is quantitative, exact, and reproducible. The modern interpretation of the reactions in nonaqueous solvents started in the last century, but several inconsistencies and unsolved problems can be found in the literature. The acid base theories of Brønsted Lowry and Lewis as well as the so-called solvent theory are outlined first, then the promoting (and leveling) and the differentiating effects are discussed on the basis of the hydrogen-bond concept. Emphasis is put on the properties of formic acid and acetic anhydride since their importance is increasing.

  4. Potentiometric Acid-Base Titrations with Activated Graphite Electrodes

    NASA Astrophysics Data System (ADS)

    Riyazuddin, P.; Devika, D.

    1997-10-01

    Dry cell graphite (DCG) electrodes activated with potassium permanganate are employed as potentiometric indicator electrodes for acid-base titrations. Special attention is given to an indicator probe comprising activated DCG-non-activiated DCG electrode couple. This combination also proves suitable for the titration of strong or weak acids.

  5. A Closer Look at Acid-Base Olfactory Titrations

    ERIC Educational Resources Information Center

    Neppel, Kerry; Oliver-Hoyo, Maria T.; Queen, Connie; Reed, Nicole

    2005-01-01

    Olfactory titrations using raw onions and eugenol as acid-base indicators are reported. An in-depth investigation on olfactory titrations is presented to include requirements for potential olfactory indicators and protocols for using garlic, onions, and vanillin as acid-base olfactory indicators are tested.

  6. Charge characteristics of humic and fulvic acids: comparative analysis by colloid titration and potentiometric titration with continuous pK-distribution function model.

    PubMed

    Bratskaya, S; Golikov, A; Lutsenko, T; Nesterova, O; Dudarchik, V

    2008-09-01

    Charge characteristics of humic and fulvic acids of a different origin (inshore soils, peat, marine sediments, and soil (lysimetric) waters) were evaluated by means of two alternative methods - colloid titration and potentiometric titration. In order to elucidate possible limitations of the colloid titration as an express method of analysis of low content of humic substances we monitored changes in acid-base properties and charge densities of humic substances with soil depth, fractionation, and origin. We have shown that both factors - strength of acidic groups and molecular weight distribution in humic and fulvic acids - can affect the reliability of colloid titration. Due to deviations from 1:1 stoichiometry in interactions of humic substances with polymeric cationic titrant, the colloid titration can underestimate total acidity (charge density) of humic substances with domination of weak acidic functional groups (pK>6) and high content of the fractions with molecular weight below 1kDa.

  7. Simultaneous determination of equivalence volumes and acid dissociation constants from potentiometric titration data.

    PubMed

    Papanastasiou, G; Ziogas, I

    1995-06-01

    New iterative methods for analysis of potentiometric titration data of (a) mixtures of weak monoprotic acids with their conjugate bases, (b) solutions of polyprotic (di- and triprotic) acids, and (c) mixtures of two diprotic acids are presented. These methods, using data exclusively resulting from the acidic region of the titration curve permits the accurate determination of the analytical concentration of one or more acids even if the titration is stopped well before the end point of the titration. For the titration of a solution containing a conjugate acid/base pair, the proposed procedure enables the extraction of the initial composition of the mixture, as well as the dissociation constant of the concerned acid. Thus, it is possible by this type of analysis to distinguish whether a weak acid has been contaminated by a strong base and define the extent of the contamination. On the other hand, for the titration of polyprotic acids, the proposed approach enables the extraction of the accurate values of the equivalence volume and the dissociation constants K(i) even when the ionization stages overlap. Finally, for the titration of a mixture of two diprotic acids the proposed procedure enables the determination of the composition of the mixture even if the sum of the concentrations of the acids is not known. This method can be used in the analysis of solutions containing two diastereoisomeric forms of a weak diprotic acid. The test of the proposed procedures by means of ideal and Monte Carlo simulated data revealed that these methods are fairly applicable even when the titration data are considerably obscured by 'noise' or contain an important systematic error. The proposed procedures were also successfully applied to experimental titration data.

  8. Sensitivity of the acid-base properties of clays to the methods of preparation and measurement. 2. Evidence from continuous potentiometric titrations.

    PubMed

    Duc, Myriam; Gaboriaud, Fabien; Thomas, Fabien

    2005-09-01

    The effects of experimental procedures on the acid-base consumption titration curves of montmorillonite suspension were studied using continuous potentiometric titration. For that purpose, the hysteresis amplitudes between the acid and base branches were found to be useful to systematically evaluate the impacts of storage conditions (wet or dried), the atmosphere in titration reactor, the solid-liquid ratio, the time interval between successive increments, and the ionic strength. In the case of storage conditions, the increase of the hysteresis was significantly higher for longer storage of clay in suspension and drying procedures compared to "fresh" clay suspension. The titration carried out under air demonstrated carbonate contamination that could only be cancelled by performing experiments under inert gas. Interestingly, the increase of the time intervals between successive increments of titrant strongly emphasized the amplitude of hysteresis, which could be correlated with the slow kinetic process specifically observed for acid addition in acid media. Thus, such kinetic behavior is probably associated with dissolution processes of clay particles. However, the resulting curves recorded at different ionic strengths under optimized conditions did not show the common intersection point required to define point of zero charge. Nevertheless, the ionic strength dependence of the point of zero net proton charge suggested that the point of zero charge of sodic montmorillonite could be estimated as lower than 5.

  9. Titration of strong and weak acids by sequential injection analysis technique.

    PubMed

    Maskula, S; Nyman, J; Ivaska, A

    2000-05-31

    A sequential injection analysis (SIA) titration method has been developed for acid-base titrations. Strong and weak acids in different concentration ranges have been titrated with a strong base. The method is based on sequential aspiration of an acidic sample zone and only one zone of the base into a carrier stream of distilled water. On their way to the detector, the sample and the reagent zones are partially mixed due to the dispersion and thereby the base is partially neutralised by the acid. The base zone contains the indicator. An LED-spectrophotometer is used as detector. It senses the colour of the unneutralised base and the signal is recorded as a typical SIA peak. The peak area of the unreacted base was found to be proportional to the logarithm of the acid concentration. Calibration curves with good linearity were obtained for a strong acid in the concentration ranges of 10(-4)-10(-2) and 0.1-3 M. Automatic sample dilution was implemented when sulphuric acid at concentration of 6-13 M was titrated. For a weak acid, i.e. acetic acid, a linear calibration curve was obtained in the range of 3x10(-4)-8x10(-2) M. By changing the volumes of the injected sample and the reagent, different acids as well as different concentration ranges of the acids can be titrated without any other adjustments in the SIA manifold or the titration protocol. PMID:18967966

  10. Titration of strong and weak acids by sequential injection analysis technique.

    PubMed

    Maskula, S; Nyman, J; Ivaska, A

    2000-05-31

    A sequential injection analysis (SIA) titration method has been developed for acid-base titrations. Strong and weak acids in different concentration ranges have been titrated with a strong base. The method is based on sequential aspiration of an acidic sample zone and only one zone of the base into a carrier stream of distilled water. On their way to the detector, the sample and the reagent zones are partially mixed due to the dispersion and thereby the base is partially neutralised by the acid. The base zone contains the indicator. An LED-spectrophotometer is used as detector. It senses the colour of the unneutralised base and the signal is recorded as a typical SIA peak. The peak area of the unreacted base was found to be proportional to the logarithm of the acid concentration. Calibration curves with good linearity were obtained for a strong acid in the concentration ranges of 10(-4)-10(-2) and 0.1-3 M. Automatic sample dilution was implemented when sulphuric acid at concentration of 6-13 M was titrated. For a weak acid, i.e. acetic acid, a linear calibration curve was obtained in the range of 3x10(-4)-8x10(-2) M. By changing the volumes of the injected sample and the reagent, different acids as well as different concentration ranges of the acids can be titrated without any other adjustments in the SIA manifold or the titration protocol.

  11. Potentiometric titration and equivalent weight of humic acid

    USGS Publications Warehouse

    Pommer, A.M.; Breger, I.A.

    1960-01-01

    The "acid nature" of humic acid has been controversial for many years. Some investigators claim that humic acid is a true weak acid, while others feel that its behaviour during potentiometric titration can be accounted for by colloidal adsorption of hydrogen ions. The acid character of humic acid has been reinvestigated using newly-derived relationships for the titration of weak acids with strong base. Re-interpreting the potentiometric titration data published by Thiele and Kettner in 1953, it was found that Merck humic acid behaves as a weak polyelectrolytic acid having an equivalent weight of 150, a pKa of 6.8 to 7.0, and a titration exponent of about 4.8. Interdretation of similar data pertaining to the titration of phenol-formaldehyde and pyrogallol-formaldehyde resins, considered to be analogs for humic acid by Thiele and Kettner, leads to the conclusion that it is not possible to differentiate between adsorption and acid-base reaction for these substances. ?? 1960.

  12. Going Beyond, Going Further: The Preparation of Acid-Base Titration Curves.

    ERIC Educational Resources Information Center

    McClendon, Michael

    1984-01-01

    Background information, list of materials needed, and procedures used are provided for a simple technique for generating mechanically plotted acid-base titration curves. The method is suitable for second-year high school chemistry students. (JN)

  13. Acid-Base Titration of (S)-Aspartic Acid: A Circular Dichroism Spectrophotometry Experiment

    NASA Astrophysics Data System (ADS)

    Cavaleiro, Ana M. V.; Pedrosa de Jesus, Júlio D.

    2000-09-01

    The magnitude of the circular dichroism of (S)-aspartic acid in aqueous solutions at a fixed wavelength varies with the addition of strong base. This laboratory experiment consists of the circular dichroism spectrophotometric acid-base titration of (S)-aspartic acid in dilute aqueous solutions, and the use of the resulting data to determine the ionization constant of the protonated amino group. The work familiarizes students with circular dichroism and illustrates the possibility of performing titrations using a less usual instrumental method of following the course of a reaction. It shows the use of a chiroptical property in the determination of the concentration in solution of an optically active molecule, and exemplifies the use of a spectrophotometric titration in the determination of an ionization constant.

  14. A General Simulator for Acid-Base Titrations

    NASA Astrophysics Data System (ADS)

    de Levie, Robert

    1999-07-01

    General formal expressions are provided to facilitate the automatic computer calculation of acid-base titration curves of arbitrary mixtures of acids, bases, and salts, without and with activity corrections based on the Davies equation. Explicit relations are also given for the buffer strength of mixtures of acids, bases, and salts.

  15. Process monitored spectrophotometric titration coupled with chemometrics for simultaneous determination of mixtures of weak acids.

    PubMed

    Liao, Lifu; Yang, Jing; Yuan, Jintao

    2007-05-15

    A new spectrophotometric titration method coupled with chemometrics for the simultaneous determination of mixtures of weak acids has been developed. In this method, the titrant is a mixture of sodium hydroxide and an acid-base indicator, and the indicator is used to monitor the titration process. In a process of titration, both the added volume of titrant and the solution acidity at each titration point can be obtained simultaneously from an absorption spectrum by least square algorithm, and then the concentration of each component in the mixture can be obtained from the titration curves by principal component regression. The method only needs the information of absorbance spectra to obtain the analytical results, and is free of volumetric measurements. The analyses are independent of titration end point and do not need the accurate values of dissociation constants of the indicator and the acids. The method has been applied to the simultaneous determination of the mixtures of benzoic acid and salicylic acid, and the mixtures of phenol, o-chlorophenol and p-chlorophenol with satisfactory results.

  16. Titration of phosphonic acid derivatives in mixtures.

    PubMed

    Wittmann, Z

    1980-01-01

    An analytical procedure is described for the determination of the weak acids phosphonomethyliminodiacetic acid and phosphonomethyliminoacetic acid in their mixtures, and the dissociation constants of phosphonomethyliminoacetic acid are reported.

  17. Rapid method for SV40 titration.

    PubMed

    Drayman, Nir; Kler, Slava; Ben-nun-Shaul, Orly; Oppenheim, Ariella

    2010-03-01

    SV40 titer is determined traditionally by the conventional plaque assay. Plaques appear after several rounds of infection and the assay takes around two weeks, which may delay research. A simpler assay was developed, based on detection of T-antigen in the infected cells by flow cytometry. Cells grown in 6-well plates are infected with serial dilutions of the viral stock, harvested 48h post-infection, stained and analyzed for T-antigen using a flow cytometer. The viral titer is calculated based on the percentage of T-antigen positive cells. The procedure is accomplished in 2 days. Unexpectedly we found that titers on different permissive African Green Monkey kidney cell lines were consistently different, suggesting variable susceptibility to SV40 infection. The method described, optimized for SV40 titration, may be adapted readily to other viruses.

  18. Acid-base titrations for polyacids: Significance of the pK sub a and parameters in the Kern equation

    NASA Technical Reports Server (NTRS)

    Meites, L.

    1978-01-01

    A new method is suggested for calculating the dissociation constants of polyvalent acids, especially polymeric acids. In qualitative form the most significant characteristics of the titration curves are demonstrated and identified which are obtained when titrating the solutions of such acids with a standard base potentiometrically.

  19. A non-linear least-squares approach to the refinement of all parameters involved in acid-base titrations.

    PubMed

    Arena, G; Rizzarelli, E; Sammartano, S; Rigano, C

    1979-01-01

    A non-linear least-squares computer program has been written for the refinement of the parameters involved in potentiometric acid-base titrations. The program ACBA (ACid-BAse titrations) is applicable under quite general conditions to solutions containing one or more acids or bases. The method of refinement used gives the program several advantages over the other programs described previously.

  20. Microbiological titration of proteins and of single amino acid content in biological materials without purification and hydrolysis.

    PubMed

    Puppo, S; Morpurgo, G; Nardi, S; Conti, G

    1978-04-01

    A method is described for the microbiological determination of the protein content of biological materials. This method can also be adopted to titrate the concentration of a single amino acid in the protein and has the following advantages: (1) titration can be done without purification and hydrolysis of proteins; (2) the titration graph is a straight line between 25 and 800 microgram/ml; (3) protein values agree with those obtained using the Kjeldhal method; and (4) each mutant requiring one amino acid may be used to titrate the concentration of a single amino acid of the protein. The leucine content of various kinds of flour was measured with this system.

  1. Differential titration of bases in glacial acetic acid.

    PubMed

    Castellano, T; Medwick, T; Shinkai, J H; Bailey, L

    1981-01-01

    A study of bases in acetic acid and their differential titration was carried out. The overall basicity constants for 20 bases were measured in acetic acid, and the differential titration of five binary mixtures of variable delta pKb values in acetic acid was followed using a glass electrode-modified calomel electrode system. Agreement with literature values was good. A leveling diagram was constructed that indicated that bases stronger than aqueous pKb 10 are leveled to an acetous pKb 5.69, whereas weaker bases are not leveled but instead exhibit their own intrinsic basicity, with the acetous pKb to aqueous pKb values being linearly related (slope 1.18, correlation coefficient 0.962). A minimum acetous delta pKb of four units is required for the satisfactory differential titration of two bases in acetic acid.

  2. Ion-selective electrodes in potentiometric titrations; a new method for processing and evaluating titration data.

    PubMed

    Granholm, Kim; Sokalski, Tomasz; Lewenstam, Andrzej; Ivaska, Ari

    2015-08-12

    A new method to convert the potential of an ion-selective electrode to concentration or activity in potentiometric titration is proposed. The advantage of this method is that the electrode standard potential and the slope of the calibration curve do not have to be known. Instead two activities on the titration curve have to be estimated e.g. the starting activity before the titration begins and the activity at the end of the titration in the presence of large excess of titrant. This new method is beneficial when the analyte is in a complexed matrix or in a harsh environment which affects the properties of the electrode and the traditional calibration procedure with standard solutions cannot be used. The new method was implemented both in a method of linearization based on the Grans's plot and in determination of the stability constant of a complex and the concentration of the complexing ligand in the sample. The new method gave accurate results when using titrations data from experiments with samples of known composition and with real industrial harsh black liquor sample. A complexometric titration model was also developed.

  3. New method for titration of virus infectivity by immunostaining.

    PubMed

    Usuba, O; Schulman, J L; Deatly, A M; Bona, C A; Moran, T M

    1990-01-01

    We have developed a new method for titration of viruses utilizing automated microtiter technology. Compared to existing methods such as plaque assay or hemagglutination titration of influenza virus, the new method offers distinct advantages in terms of time and effort. In addition because multiple replicates can easily be employed accuracy can be increased.

  4. Ultrasonic and densimetric titration applied for acid-base reactions.

    PubMed

    Burakowski, Andrzej; Gliński, Jacek

    2014-01-01

    Classical acoustic acid-base titration was monitored using sound speed and density measurements. Plots of these parameters, as well as of the adiabatic compressibility coefficient calculated from them, exhibit changes with the volume of added titrant. Compressibility changes can be explained and quantitatively predicted theoretically in terms of Pasynski theory of non-compressible hydrates combined with that of the additivity of the hydration numbers with the amount and type of ions and molecules present in solution. It also seems that this development could be applied in chemical engineering for monitoring the course of chemical processes, since the applied experimental methods can be carried out almost independently on the medium under test (harmful, aggressive, etc.).

  5. On the accuracy of acid-base determinations from potentiometric titrations using only a few points from the titration curve.

    PubMed

    Olin, A; Wallén, B

    1977-05-01

    There are several procedures which use only a few points on the titration curve for the calculation of equivalence volumes in acid-base titrations. The accuracy of such determinations will depend on the positions of the points on the titration curve. The effects of errors in the stability constants and in the pH measurements on the accuracy of the analysis have been considered, and the results are used to establish the conditions under which these errors are minimized.

  6. Using Spreadsheets to Produce Acid-Base Titration Curves.

    ERIC Educational Resources Information Center

    Cawley, Martin James; Parkinson, John

    1995-01-01

    Describes two spreadsheets for producing acid-base titration curves, one uses relatively simple cell formulae that can be written into the spreadsheet by inexperienced students and the second uses more complex formulae that are best written by the teacher. (JRH)

  7. A fully automatic system for acid-base coulometric titrations.

    PubMed

    Cladera, A; Caro, A; Estela, J M; Cerdà, V

    1990-01-01

    An automatic system for acid-base titrations by electrogeneration of H(+) and OH(-) ions, with potentiometric end-point detection, was developed. The system includes a PC-compatible computer for instrumental control, data acquisition and processing, which allows up to 13 samples to be analysed sequentially with no human intervention.The system performance was tested on the titration of standard solutions, which it carried out with low errors and RSD. It was subsequently applied to the analysis of various samples of environmental and nutritional interest, specifically waters, soft drinks and wines.

  8. Ascorbic Acid as a Standard for Iodometric Titrations. An Analytical Experiment for General Chemistry

    NASA Astrophysics Data System (ADS)

    Silva, Cesar R.; Simoni, Jose A.; Collins, Carol H.; Volpe, Pedro L. O.

    1999-10-01

    Ascorbic acid is suggested as the weighable compound for the standardization of iodine solutions in an analytical experiment in general chemistry. The experiment involves an iodometric titration in which iodine reacts with ascorbic acid, oxidizing it to dehydroascorbic acid. The redox titration endpoint is determined by the first iodine excess that is complexed with starch, giving a deep blue-violet color. The results of the titration of iodine solution using ascorbic acid as a calibration standard were compared with the results acquired by the classic method using a standardized solution of sodium thiosulfate. The standardization of the iodine solution using ascorbic acid was accurate and precise, with the advantages of saving time and avoiding mistakes due to solution preparation. The colorless ascorbic acid solution gives a very clear and sharp titration end point with starch. It was shown by thermogravimetric analysis that ascorbic acid can be dried at 393 K for 2 h without decomposition. This experiment allows general chemistry students to perform an iodometric titration during a single laboratory period, determining with precision the content of vitamin C in pharmaceutical formulations.

  9. Titration of fatty acids solubilized in cationic and anionic micelles. Calorimetry and thermodynamic modeling.

    PubMed

    Söderman, Olle; Jönsson, Bengt; Olofsson, Gerd

    2006-02-23

    The electrostatic properties of charged surfactant micelles are investigated through titrations of fatty acid probes solubilized in the micelles. The titration process is followed by means of calorimetric measurements and by determining the pH values as a function of added base. This approach yields a complete thermodynamic description of the titration process. In particular, we find that the process is endothermic at 298 K. This is contrary to the titration of carboxylic acids in water, where DeltaH is approximately 0. To identify the main effect underlying the difference in DeltaH between titration in a micelle and water, a thermodynamic model has been developed which focuses on the transfer properties of charged and uncharged species from bulk water to the surface of a micelle and which incorporates a dielectric discontinuity at the micellar surface. The model relies on the use of the Poisson-Boltzmann equation which is solved using a finite element method. Experimental results and the model calculations imply that the dielectric discontinuity at (or near) the micellar surface plays a major role and hence must be included when analyzing the titration behavior of an acid functionality at the surface of a charged micelle.

  10. Micellar acid-base potentiometric titrations of weak acidic and/or insoluble drugs.

    PubMed

    Gerakis, A M; Koupparis, M A; Efstathiou, C E

    1993-01-01

    The effect of various surfactants [the cationics cetyl trimethyl ammonium bromide (CTAB) and cetyl pyridinium chloride (CPC), the anionic sodium dodecyl sulphate (SDS), and the nonionic polysorbate 80 (Tween 80)] on the solubility and ionization constant of some sparingly soluble weak acids of pharmaceutical interest was studied. Benzoic acid (and its 3-methyl-, 3-nitro-, and 4-tert-butyl-derivatives), acetylsalicylic acid, naproxen and iopanoic acid were chosen as model examples. Precise and accurate acid-base titrations in micellar systems were made feasible using a microcomputer-controlled titrator. The response curve, response time and potential drift of the glass electrode in the micellar systems were examined. The cationics CTAB and CPC were found to increase considerably the ionization constant of the weak acids (delta pKa ranged from -0.21 to -3.57), while the anionic SDS showed negligible effect and the nonionic Tween 80 generally decreased the ionization constants. The solubility of the acids in aqueous micellar and acidified micellar solutions was studied spectrophotometrically and it was found increased in all cases. Acetylsalicylic acid, naproxen, benzoic acid and iopanoic acid could be easily determined in raw material and some of them in pharmaceutical preparations by direct titration in CTAB-micellar system instead of using the traditional non-aqueous or back titrimetry. Precisions of 0.3-4.3% RSD and good correlation with the official tedious methods were obtained. The interference study of some excipients showed that a preliminary test should be carried out before the assay of formulations.

  11. An on-line potentiometric sequential injection titration process analyser for the determination of acetic acid.

    PubMed

    van Staden, J F; Mashamba, Mulalo G; Stefan, Raluca I

    2002-09-01

    An on-line potentiometric sequential injection titration process analyser for the determination of acetic acid is proposed. A solution of 0.1 mol L(-1) sodium chloride is used as carrier. Titration is achieved by aspirating acetic acid samples between two strong base-zone volumes into a holding coil and by channelling the stack of well-defined zones with flow reversal through a reaction coil to a potentiometric sensor where the peak widths were measured. A linear relationship between peak width and logarithm of the acid concentration was obtained in the range 1-9 g/100 mL. Vinegar samples were analysed without any sample pre-treatment. The method has a relative standard deviation of 0.4% with a sample frequency of 28 samples per hour. The results revealed good agreement between the proposed sequential injection and an automated batch titration method.

  12. Acid-base titrations by stepwise addition of equal volumes of titrant with special reference to automatic titrations-III Presentation of a fully automatic titration apparatus and of results supporting the theories given in the preceding parts.

    PubMed

    Pehrsson, L; Ingman, F

    1977-02-01

    This paper forms Part III of a series in which the first two parts describe methods for evaluating titrations performed by stepwise addition of equal volumes of titrant. The great advantage of these methods is that they do not require an accurate calibration of the electrode system. This property makes the methods very suitable for routine work. e.g., in automatic analysis. An apparatus for performing such titrations automatically is presented. Further, results of titrations of monoprotic acids, a diprotic acid, an ampholyte, a mixture of an acid with its conjugate base, and mixtures of two acids with a small difference between the stability constants are given. Most of these titrations cannot be evaluated by the Gran or Hofstee methods but yield results having errors of the order of 0.1% if the methods proposed in Parts I and II of this series are employed. The advantages of the method of stepwise addition of equal volumes of titrant combined with the proposed evaluation methods, in comparison with common methods such as titration to a preset pH, are that all the data are used in the evaluation, permitting a statistical treatment and giving better possibilities for tracing systematic errors.

  13. Determination of berkelium by the method of spectrophotometric titration

    SciTech Connect

    Frolova, L.M.; Vityutnev, V.M.; Vasil'ev, V.M.

    1987-01-01

    The method that the authors propose consists of the following: berkelium is oxidized electrochemically, spectrophotometric titration of berkelium(IV) by a solution of the reducing agent is performed, and the amount of berkelium(IV) is determined according to the volume of the titrant, and considering the degree of oxidation of berkelium(III) to berkelium(IV), the total berkelium content in the sample is also determined. In this case the necessity for preliminary determination of the molar extinction coefficient of berkelium(IV) under the experimental conditions falls away. Moreover, the radiometric method of determining the berkelium content is not used. Successful titration requires selection of a reagent which, on the one hand, would rapidly reduce berkelium(VI), but on the other hand, neither itself nor the reaction products would interfere with the measurement of the optical density of berkelium(IV). As is well known, berkelium(IV) is quantitatively and rapidly reduced by hydrogen peroxide (10, 11), hydroxylamine (11), and nitrous acid (9). After preliminary experiments, they selected hydrogen peroxide and sodium nitrite as the titrants.

  14. Trace analysis of acids and bases by conductometric titration with multiparametric non-linear regression.

    PubMed

    Coelho, Lúcia H G; Gutz, Ivano G R

    2006-03-15

    A chemometric method for analysis of conductometric titration data was introduced to extend its applicability to lower concentrations and more complex acid-base systems. Auxiliary pH measurements were made during the titration to assist the calculation of the distribution of protonable species on base of known or guessed equilibrium constants. Conductivity values of each ionized or ionizable species possibly present in the sample were introduced in a general equation where the only unknown parameters were the total concentrations of (conjugated) bases and of strong electrolytes not involved in acid-base equilibria. All these concentrations were adjusted by a multiparametric nonlinear regression (NLR) method, based on the Levenberg-Marquardt algorithm. This first conductometric titration method with NLR analysis (CT-NLR) was successfully applied to simulated conductometric titration data and to synthetic samples with multiple components at concentrations as low as those found in rainwater (approximately 10 micromol L(-1)). It was possible to resolve and quantify mixtures containing a strong acid, formic acid, acetic acid, ammonium ion, bicarbonate and inert electrolyte with accuracy of 5% or better.

  15. Macromolecular competition titration method accessing thermodynamics of the unmodified macromolecule-ligand interactions through spectroscopic titrations of fluorescent analogs.

    PubMed

    Bujalowski, Wlodzimierz; Jezewska, Maria J

    2011-01-01

    Analysis of thermodynamically rigorous binding isotherms provides fundamental information about the energetics of the ligand-macromolecule interactions and often an invaluable insight about the structure of the formed complexes. The Macromolecular Competition Titration (MCT) method enables one to quantitatively obtain interaction parameters of protein-nucleic acid interactions, which may not be available by other methods, particularly for the unmodified long polymer lattices and specific nucleic acid substrates, if the binding is not accompanied by adequate spectroscopic signal changes. The method can be applied using different fluorescent nucleic acids or fluorophores, although the etheno-derivatives of nucleic acid are especially suitable as they are relatively easy to prepare, have significant blue fluorescence, their excitation band lies far from the protein absorption spectrum, and the modification eliminates the possibility of base pairing with other nucleic acids. The MCT method is not limited to the specific size of the reference nucleic acid. Particularly, a simple analysis of the competition titration experiments is described in which the fluorescent, short fragment of nucleic acid, spanning the exact site-size of the protein-nucleic acid complex, and binding with only a 1:1 stoichiometry to the protein, is used as a reference macromolecule. Although the MCT method is predominantly discussed as applied to studying protein-nucleic acid interactions, it can generally be applied to any ligand-macromolecule system by monitoring the association reaction using the spectroscopic signal originating from the reference macromolecule in the presence of the competing macromolecule, whose interaction parameters with the ligand are to be determined.

  16. Titratable acidity of beverages influences salivary pH recovery.

    PubMed

    Tenuta, Livia Maria Andaló; Fernández, Constanza Estefany; Brandão, Ana Carolina Siqueira; Cury, Jaime Aparecido

    2015-01-01

    A low pH and a high titratable acidity of juices and cola-based beverages are relevant factors that contribute to dental erosion, but the relative importance of these properties to maintain salivary pH at demineralizing levels for long periods of time after drinking is unknown. In this crossover study conducted in vivo, orange juice, a cola-based soft drink, and a 10% sucrose solution (negative control) were tested. These drinks differ in terms of their pH (3.5 ± 0.04, 2.5 ± 0.05, and 5.9 ± 0.1, respectively) and titratable acidity (3.17 ± 0.06, 0.57 ± 0.04 and < 0.005 mmols OH- to reach pH 5.5, respectively). Eight volunteers with a normal salivary flow rate and buffering capacity kept 15 mL of each beverage in their mouth for 10 s, expectorated it, and their saliva was collected after 15, 30, 45, 60, 90, and 120 s. The salivary pH, determined using a mini pH electrode, returned to the baseline value at 30 s after expectoration of the cola-based soft drink, but only at 90 s after expectoration of the orange juice. The salivary pH increased to greater than 5.5 at 15 s after expectoration of the cola drink and at 30 s after expectoration of the orange juice. These findings suggest that the titratable acidity of a beverage influences salivary pH values after drinking acidic beverages more than the beverage pH.

  17. Using spectrophotometric titrations to characterize humic acid reactivity at environmental concentrations.

    PubMed

    Janot, Noémie; Reiller, Pascal E; Korshin, Gregory V; Benedetti, Marc F

    2010-09-01

    Potentiometric titration is a common method to characterize dissolved organic matter (DOM) reactivity. Because of the sensitivity of pH electrodes, it is necessary to work with very high DOM (>1 g/L) concentrations that are unrealistic compared to those found in natural waters (0.1 to 100 mg/L). To obtain proton binding data for concentrations closer to environmental values, spectroscopic titration methodology is a viable alternative to traditional potentiometric titrations. Spectrophotometric titrations and UV-visible spectra of a diluted solution of purified Aldrich humic acid (5 mgDOC/L) are used to estimate changes in proton binding moieties as function of pH and ionic strength after calculation of differential absorbance spectra variations. After electrostatic correction of spectrophotometric data, there is a linear operational correlation between spectrophotometric and potentiometric data which can be used as a transfer function between the two properties. Spectrophotometric titrations are then used to determine the changes of humic acid protonation after adsorption onto alpha-alumina.

  18. Determination of free acidic and alkaline residues of protein via moving reaction boundary titration in microdevice electrophoresis.

    PubMed

    Wang, Hou-yu; Li, Si; Tang, Yun-yun; Dong, Jing-yu; Fan, Liu-yin; Cao, Cheng-xi

    2013-06-21

    As two important physico-chemical parameters, the acidic and alkaline residues of protein are of evident significance for the evaluation of protein properties and the design of relevant separation and analysis. However, there is still no electrophoretic method used for the direct detection of free acidic and alkaline residues of protein. Herein, we developed the concepts of moving reaction boundary (MRB) and MRB titration, relevant MRB titration theory, and the method of microdevice electrophoresis for the determination of free acidic and alkaline residues of protein. In the MRB titration, the boundary was created with acid or alkali and target protein immobilized via highly cross-linked polyacrylamide gel (PAG). It was theoretically revealed that the number of free acidic or alkaline residues of protein was as a function of MRB displacement in the electrophoretic titration system. As a proof of concept, seven model proteins were chosen for the determination of acidic or alkaline residues of protein via MRB titration. The results showed that the numbers of free acidic and alkaline residues of proteins detected were in good agreement with those obtained from the relevant amino sequences in the NCBI database, demonstrating the feasibility of the developed concept, theory and technique. The general methodology of MRB titration has potential application for inexpensive, facilitative and informative protein structure analysis of free acidic or alkaline residues of protein.

  19. Interaction between bisphenol A and tannic Acid: Spectroscopic titration approach

    NASA Astrophysics Data System (ADS)

    Omoike, Anselm; Brandt, Benjamin

    2011-06-01

    The interaction between tannic acid (TA) and bisphenol A (BPA), an endocrine disruptor, was studied by absorption and fluorescence titration techniques. The binding constants and corresponding thermodynamic parameters at different temperatures (294, 296, 298, 300 and 303 K) were determined. The intrinsic fluorescence of BPA was strongly quenched by TA and the quenching mechanism is attributed to static quenching. The thermodynamic data revealed that the formation of TA-BPA complex was exothermic, entropic-driven, and spontaneous. Furthermore, hydrogen and van der Waals interactions seem to be the major driving forces for the formation of the nonfluorescent TA-BPA complex.

  20. Interaction between bisphenol A and tannic acid: spectroscopic titration approach.

    PubMed

    Omoike, Anselm; Brandt, Benjamin

    2011-06-01

    The interaction between tannic acid (TA) and bisphenol A (BPA), an endocrine disruptor, was studied by absorption and fluorescence titration techniques. The binding constants and corresponding thermodynamic parameters at different temperatures (294, 296, 298, 300 and 303 K) were determined. The intrinsic fluorescence of BPA was strongly quenched by TA and the quenching mechanism is attributed to static quenching. The thermodynamic data revealed that the formation of TA-BPA complex was exothermic, entropic-driven, and spontaneous. Furthermore, hydrogen and van der Waals interactions seem to be the major driving forces for the formation of the nonfluorescent TA-BPA complex.

  1. Error assessment in recombinant baculovirus titration: evaluation of different methods.

    PubMed

    Roldão, António; Oliveira, Rui; Carrondo, Manuel J T; Alves, Paula M

    2009-07-01

    The success of baculovirus/insect cells system in heterologous protein expression depends on the robustness and efficiency of the production workflow. It is essential that process parameters are controlled and include as little variability as possible. The multiplicity of infection (MOI) is the most critical factor since irreproducible MOIs caused by inaccurate estimation of viral titers hinder batch consistency and process optimization. This lack of accuracy is related to intrinsic characteristics of the method such as the inability to distinguish between infectious and non-infectious baculovirus. In this study, several methods for baculovirus titration were compared. The most critical issues identified were the incubation time and cell concentration at the time of infection. These variables influence strongly the accuracy of titers and must be defined for optimal performance of the titration method. Although the standard errors of the methods varied significantly (7-36%), titers were within the same order of magnitude; thus, viral titers can be considered independent of the method of titration. A cost analysis of the baculovirus titration methods used in this study showed that the alamarblue, real time Q-PCR and plaque assays were the most expensive techniques. The remaining methods cost on average 75% less than the former methods. Based on the cost, time and error analysis undertaken in this study, the end-point dilution assay, microculture tetrazolium assay and flow cytometric assay were found to be the techniques that combine all these three main factors better. Nevertheless, it is always recommended to confirm the accuracy of the titration either by comparison with a well characterized baculovirus reference stock or by titration using two different methods and verification of the variability of results.

  2. Sequential injection redox or acid-base titration for determination of ascorbic acid or acetic acid.

    PubMed

    Lenghor, Narong; Jakmunee, Jaroon; Vilen, Michael; Sara, Rolf; Christian, Gary D; Grudpan, Kate

    2002-12-01

    Two sequential injection titration systems with spectrophotometric detection have been developed. The first system for determination of ascorbic acid was based on redox reaction between ascorbic acid and permanganate in an acidic medium and lead to a decrease in color intensity of permanganate, monitored at 525 nm. A linear dependence of peak area obtained with ascorbic acid concentration up to 1200 mg l(-1) was achieved. The relative standard deviation for 11 replicate determinations of 400 mg l(-1) ascorbic acid was 2.9%. The second system, for acetic acid determination, was based on acid-base titration of acetic acid with sodium hydroxide using phenolphthalein as an indicator. The decrease in color intensity of the indicator was proportional to the acid content. A linear calibration graph in the range of 2-8% w v(-1) of acetic acid with a relative standard deviation of 4.8% (5.0% w v(-1) acetic acid, n=11) was obtained. Sample throughputs of 60 h(-1) were achieved for both systems. The systems were successfully applied for the assays of ascorbic acid in vitamin C tablets and acetic acid content in vinegars, respectively.

  3. Rapid determination of the equivalence volume in potentiometric acid-base titrations to a preset pH-I Theory and applications.

    PubMed

    Ivaska, A

    1974-06-01

    A new approach to shorten the time needed for an acid-base titration has been made. The method developed is based on the equation for acid-base titrations derived by Ingman and Still. The equation is transformed into such a form that only one titration point is needed to calculate the equivalence volume when the titration is carried out to a preset pH which can be chosen according to the experimental conditions. The method is used for titration of acetic acid, log K(H)(HA) = 4.65, hydroxylammonium ion, log K(H)(HA) approximately 6.2, and boric acid, log K(H)(HA) approximately 9.1, with an error of 0.1-0.5%. In titration of hydrogen ascorbate ion, log K(H)(HA) approximately 11.3, the error obtained was about 0.3-2%.

  4. Equivalence-point electromigration acid-base titration via moving neutralization boundary electrophoresis.

    PubMed

    Yang, Qing; Fan, Liu-Yin; Huang, Shan-Sheng; Zhang, Wei; Cao, Cheng-Xi

    2011-04-01

    In this paper, we developed a novel method of acid-base titration, viz. the electromigration acid-base titration (EABT), via a moving neutralization boundary (MNR). With HCl and NaOH as the model strong acid and base, respectively, we conducted the experiments on the EABT via the method of moving neutralization boundary for the first time. The experiments revealed that (i) the concentration of agarose gel, the voltage used and the content of background electrolyte (KCl) had evident influence on the boundary movement; (ii) the movement length was a function of the running time under the constant acid and base concentrations; and (iii) there was a good linearity between the length and natural logarithmic concentration of HCl under the optimized conditions, and the linearity could be used to detect the concentration of acid. The experiments further manifested that (i) the RSD values of intra-day and inter-day runs were less than 1.59 and 3.76%, respectively, indicating similar precision and stability in capillary electrophoresis or HPLC; (ii) the indicators with different pK(a) values had no obvious effect on EABT, distinguishing strong influence on the judgment of equivalence-point titration in the classic one; and (iii) the constant equivalence-point titration always existed in the EABT, rather than the classic volumetric analysis. Additionally, the EABT could be put to good use for the determination of actual acid concentrations. The experimental results achieved herein showed a new general guidance for the development of classic volumetric analysis and element (e.g. nitrogen) content analysis in protein chemistry.

  5. Acid-base titration curves for acids with very small ratios of successive dissociation constants.

    PubMed

    Campbell, B H; Meites, L

    1974-02-01

    The shapes of the potentiometric acid-base titration curves obtained in the neutralizations of polyfunctional acids or bases for which each successive dissociation constant is smaller than the following one are examined. In the region 0 < < 1 (where is the fraction of the equivalent volume of reagent that has been added) the slope of the titration curve decreases as the number j of acidic or basic sites increases. The difference between the pH-values at = 0.75 and = 0.25 has (1 j)log 9 as the lower limit of its maximum value.

  6. Developing a multipoint titration method with a variable dose implementation for anaerobic digestion monitoring.

    PubMed

    Salonen, K; Leisola, M; Eerikäinen, T

    2009-01-01

    Determination of metabolites from an anaerobic digester with an acid base titration is considered as superior method for many reasons. This paper describes a practical at line compatible multipoint titration method. The titration procedure was improved by speed and data quality. A simple and novel control algorithm for estimating a variable titrant dose was derived for this purpose. This non-linear PI-controller like algorithm does not require any preliminary information from sample. Performance of this controller is superior compared to traditional linear PI-controllers. In addition, simplification for presenting polyprotic acids as a sum of multiple monoprotic acids is introduced along with a mathematical error examination. A method for inclusion of the ionic strength effect with stepwise iteration is shown. The titration model is presented with matrix notations enabling simple computation of all concentration estimates. All methods and algorithms are illustrated in the experimental part. A linear correlation better than 0.999 was obtained for both acetate and phosphate used as model compounds with slopes of 0.98 and 1.00 and average standard deviations of 0.6% and 0.8%, respectively. Furthermore, insensitivity of the presented method for overlapping buffer capacity curves was shown.

  7. Semi-automated potentiometric titration method for uranium characterization.

    PubMed

    Cristiano, B F G; Delgado, J U; da Silva, J W S; de Barros, P D; de Araújo, R M S; Lopes, R T

    2012-07-01

    The manual version of the potentiometric titration method has been used for certification and characterization of uranium compounds. In order to reduce the analysis time and the influence of the analyst, a semi-automatic version of the method was developed in the Brazilian Nuclear Energy Commission. The method was applied with traceability assured by using a potassium dichromate primary standard. The combined standard uncertainty in determining the total concentration of uranium was around 0.01%, which is suitable for uranium characterization.

  8. 2-methoxyethanol as a solvent for conductometric acid-base titrations.

    PubMed

    Schwartz, G A; Barker, B J

    1975-09-01

    Conductometric titrations of a series of organic acids in 2-methoxyethanol were performed with 1,3-diphenylguanidine (DPG) as titrant. For benzoic, p-chlorobenzoic and picric acids, phenylphenol and barbital, excellent recoveries were obtained from well-defined conductance vs. volume plots. Results for the dicarboxylic phthalic and adipic acids were also good. However, the titration curves for the difunctional salicylic and p-hydroxybenzoic acids were not clearly defined. The results are discussed and compared with cnductometric titrations of acids in other non-aqueous solvents. Several determinations of electrolyte conductance as a function of concentration revealed that, as expected, the selected substances are weak electrolytes in methoxyethanol.

  9. A Computer-Based Simulation of an Acid-Base Titration

    ERIC Educational Resources Information Center

    Boblick, John M.

    1971-01-01

    Reviews the advantages of computer simulated environments for experiments, referring in particular to acid-base titrations. Includes pre-lab instructions and a sample computer printout of a student's use of an acid-base simulation. Ten references. (PR)

  10. Acid-base titrations in nonaqueous solvents Analysis of dimethyl sulphate.

    PubMed

    Banick, W M; Francis, E C

    1966-07-01

    A nonaqueous titrimetric procedure was developed for the determination of the dimethyl sulphate, methyl hydrogen sulphate and sulphuric acid content of dimethyl sulphate samples. Methyl hydrogen sulphate and sulphuric acid are determined by a differentiating potentiometric titration in pyridine with tributylethylammonium hydroxide. Pyridine converts the dimethyl sulphate into the weakly acidic methylpyridinium methyl sulphate which does not interfere in the titration. Dimethyl sulphate is determined by reacting it with an excess of 2-dimethylaminoethanol and titrating the excess with perchloric acid. Precision and recovery data for commercial samples of dimethyl sulphate are presented.

  11. New methods to titrate EIAV-based lentiviral vectors.

    PubMed

    Martin-Rendon, Enca; White, Linda J; Olsen, Anna; Mitrophanous, Kyriacos A; Mazarakis, Nicholas D

    2002-05-01

    Ideally, gene transfer vectors used in clinical protocols should only express the gene of interest. So far most vectors have contained marker genes to aid their titration. We have used quantitative real-time PCR to titrate equine infectious anemia virus (EIAV) vectors for gene therapy applications. Viral RNA was isolated from vector preparations and analyzed in a one-step RT-PCR reaction in which reverse transcription and amplification were combined in one tube. The PCR assay of vector stocks was quantitative and linear over four orders of magnitude. In tandem, the integration efficiency of these vectors has also been determined by real-time PCR, measuring the number of vector genomes in the target cells. We have found that these methods permit reliable and sensitive titration of lentiviral vectors independent from the expression of a transgene. They also allow us to determine the integration efficiency of different vector genomes. This technology has proved very useful, especially in the absence of marker genes and where vectors express multiple genes.

  12. Semi-automated potentiometric titration method for uranium characterization.

    PubMed

    Cristiano, B F G; Delgado, J U; da Silva, J W S; de Barros, P D; de Araújo, R M S; Lopes, R T

    2012-07-01

    The manual version of the potentiometric titration method has been used for certification and characterization of uranium compounds. In order to reduce the analysis time and the influence of the analyst, a semi-automatic version of the method was developed in the Brazilian Nuclear Energy Commission. The method was applied with traceability assured by using a potassium dichromate primary standard. The combined standard uncertainty in determining the total concentration of uranium was around 0.01%, which is suitable for uranium characterization. PMID:22154105

  13. Chemical force titrations of amine- and sulfonic acid-modified poly(dimethylsiloxane).

    PubMed

    Wang, Bin; Oleschuk, Richard D; Horton, J Hugh

    2005-02-15

    Chemical force titrations-measurements of the adhesive interaction between a pair of suitably chemically modified atomic force microscopy (AFM) tip and sample surfaces as a function of pH-have been carried out for various combinations of silanol, amine, carboxylic acid, and sulfonic acid functional groups on both tip and sample. The primary surface material studied was poly(dimethylsiloxane) (PDMS). Surface modification was carried out using a plasma oxidation process to form silanol sites; further modification with amine or sulfonic acid sites was carried out by reaction of the silanol sites with the appropriate trialkoxysilane derivative. AFM tips were also modified using trialkoxysilane compounds. In the cases of tip/sample combinations with the same functional group on each, surface pK(1/2) values could be determined. In several "mixed" tip/sample combinations, a peak appeared in the titration curve midway between the surface pK(1/2) values of the tip and sample, consistent with an ionic H-bonding model for the interactions. The amine/sulfonic acid pair showed more complex behavior; the amine-terminated tip/sulfonic acid-terminated PDMS surface force titration curve consisted of two peaks centered at pH 4 and pH 8. Reversing the tip/sample pair resulted in the peak positions being shifted upward by 1.0 pH unit. The peak appearing at lower pH is assigned to electrostatic interactions between the two oppositely charged surfaces, whereas the higher pH peak is believed to arise due to ionic H-bonding interactions. AFM images show the effects on surface patterning of amine- and sulfonic acid-modified PDMS surfaces that have undergone two different oxidation methods (air plasma oxidation and Tesla coil oxidation). The surface morphologies of freshly prepared and 24 h aged air plasma oxidized PDMS are also discussed in this study.

  14. Instrumentation and automated photometric titration procedure for total acidity determination in red wine employing a multicommuted flow system.

    PubMed

    Garcia, Ausberta Jesus Cabezas; Reis, Boaventura F

    2006-01-01

    An automated procedure for photometric titration of red wine and associated instrumentation is described. The procedure was based on the flow-batch approach implemented employing multicommutation. The photometric detection was carried out using a homemade LED-based photometer. The mixing device, LED, and photodetector were attached to the titration chamber in order to form a compact and small-sized unit. The flow system comprised an automatic injector and three-way solenoid valves, which were controlled by a microcomputer through an electronic interface card. The software, written in Quick BASIC 4.5, was designed with abilities to accomplish all steps of the titration procedure including data acquisition and real-time processing to decide about the course of titration in the following step and so forth, until the titration endpoint was reached. The usefulness of the proposed titration system was demonstrated by analyzing red wine samples. When results were compared with those obtained using the AOAC reference method, no significant difference was observed at the 95% confidence level. A relative standard deviation of ca 2% (n=9) was obtained when processing a typical red wine sample containing 7.3 gl-1 total acidity expressed as tartaric acid.

  15. Instrumentation and automated photometric titration procedure for total acidity determination in red wine employing a multicommuted flow system.

    PubMed

    Garcia, Ausberta Jesus Cabezas; Reis, Boaventura F

    2006-01-01

    An automated procedure for photometric titration of red wine and associated instrumentation is described. The procedure was based on the flow-batch approach implemented employing multicommutation. The photometric detection was carried out using a homemade LED-based photometer. The mixing device, LED, and photodetector were attached to the titration chamber in order to form a compact and small-sized unit. The flow system comprised an automatic injector and three-way solenoid valves, which were controlled by a microcomputer through an electronic interface card. The software, written in Quick BASIC 4.5, was designed with abilities to accomplish all steps of the titration procedure including data acquisition and real-time processing to decide about the course of titration in the following step and so forth, until the titration endpoint was reached. The usefulness of the proposed titration system was demonstrated by analyzing red wine samples. When results were compared with those obtained using the AOAC reference method, no significant difference was observed at the 95% confidence level. A relative standard deviation of ca 2% (n=9) was obtained when processing a typical red wine sample containing 7.3 gl-1 total acidity expressed as tartaric acid. PMID:17671625

  16. Colloid Titration--A Rapid Method for the Determination of Charged Colloid.

    ERIC Educational Resources Information Center

    Ueno, Keihei; Kina, Ken'yu

    1985-01-01

    "Colloid titration" is a volumetric method for determining charged polyelectrolytes in aqueous solutions. The principle of colloid titration, reagents used in the procedure, methods of endpoint detection, preparation of reagent solutions, general procedure used, results obtained, and pH profile of colloid titration are considered. (JN)

  17. pH-Metric determination of acid values in oilseeds without titration.

    PubMed

    Kuselman, I; Tur'yan, Y I; Burenko, T; Goldfeld, I; Anisimov, B

    1999-07-01

    A new pH-metric method for determination of acid values in oilseeds without titration has been developed in the range 0.6-10 and more mg KOH/g. The method is based on a rapid (1-2 min) selective and complete extraction of free fatty acids from an oilseed test portion into a special reagent A, separation of the solution from the solid oilseed material by centrifugation or filtration, transfer of an aliquot of the solution into a pH-metric cell with reagent B for measurement of conditional pH(1)' of the formed mixture, addition of standard acid (HCl or H(2)SO(4)) and pH(2)' measurement. The reagents are non-toxic, and the method is rapid. Its metrological parameters for Soybean, Canola and Sunflower oilseeds are satisfactory for practical purposes.

  18. pH-metric solubility. 3. Dissolution titration template method for solubility determination.

    PubMed

    Avdeef, A; Berger, C M

    2001-12-01

    The main objective of this study was to develop an effective potentiometric saturation titration protocol for determining the aqueous intrinsic solubility and the solubility-pH profile of ionizable molecules, with the specific aim of overcoming incomplete dissolution conditions, while attempting to shorten the data collection time. A modern theory of dissolution kinetics (an extension of the Noyes-Whitney approach) was applied to acid-base titration experiments. A thermodynamic method was developed, based on a three-component model, to calculate interfacial, diffusion-layer, and bulk-water reactant concentrations in saturated solutions of ionizable compounds perturbed by additions of acid/base titrant, leading to partial dissolution of the solid material. Ten commercial drugs (cimetidine, diltiazem hydrochloride, enalapril maleate, metoprolol tartrate, nadolol, propoxyphene hydrochloride, quinine hydrochloride, terfenadine, trovafloxacin mesylate, and benzoic acid) were chosen to illustrate the new titration methodology. It was shown that the new method is about 10 times faster in determining equilibrium solubility constants, compared to the traditional saturation shake-flask methods.

  19. A comparison of three methods for titration of poliovirus vaccines.

    PubMed

    Bey, E; Golombick, T

    1984-10-01

    Two methods for in vitro endpoint titration of poliovirus--the roller tube and the microtitration assay--were compared with each other and with the plaque assay, using secondary vervet monkey kidney cells and Vero cells as indicators. The roller tube method is the most reliable under difficult working conditions, but is otherwise cumbersome and expensive. The microtitre method is the most economical and the plaque assay the most sensitive. By suspending freshly trypsinized indicator cells with the virus dilutions before planting, it was possible to simplify the microtitre method considerably. The sensitivity of the plaque assay was improved for Vero cells by absorbing the virus onto freshly planted monolayers. The method was scaled down to a semi-micro level by using 24-well cell culture trays. The slower rate of plaque development under a low calcium overlay medium facilitated a more accurate plaque count.

  20. Theory of titration curves-VII The properties of derivative titration curves for strong acid-strong base and other isovalent ion-combination titrations.

    PubMed

    Meites, T; Meites, L

    1970-06-01

    This paper deals with isovalent ion-combination titrations based on reactions that can be represented by the equation M(n+) + X(n-) --> MX, where the activity of the product MX is invariant throughout a titration, and with the derivative titration curves obtained by plotting d[M(+)]/dfversus f for such titrations. It describes some of the ways in which such curves can be obtained; it compares and contrasts them both with potentiometric titration curves, which resemble them in shape, and with segmented titration curves, from which they are derived; and it discusses their properties in detail.

  1. Evaluation of the 5 and 8 pH point titration methods for monitoring anaerobic digesters treating solid waste.

    PubMed

    Vannecke, T P W; Lampens, D R A; Ekama, G A; Volcke, E I P

    2015-01-01

    Simple titration methods certainly deserve consideration for on-site routine monitoring of volatile fatty acid (VFA) concentration and alkalinity during anaerobic digestion (AD), because of their simplicity, speed and cost-effectiveness. In this study, the 5 and 8 pH point titration methods for measuring the VFA concentration and carbonate system alkalinity (H2CO3*-alkalinity) were assessed and compared. For this purpose, synthetic solutions with known H2CO3*-alkalinity and VFA concentration as well as samples from anaerobic digesters treating three different kind of solid wastes were analysed. The results of these two related titration methods were verified with photometric and high-pressure liquid chromatography measurements. It was shown that photometric measurements lead to overestimations of the VFA concentration in the case of coloured samples. In contrast, the 5 pH point titration method provides an accurate estimation of the VFA concentration, clearly corresponding with the true value. Concerning the H2CO3*-alkalinity, the most accurate and precise estimations, showing very similar results for repeated measurements, were obtained using the 8 pH point titration. Overall, it was concluded that the 5 pH point titration method is the preferred method for the practical monitoring of AD of solid wastes due to its robustness, cost efficiency and user-friendliness.

  2. Automated titration method for use on blended asphalts

    DOEpatents

    Pauli, Adam T.; Robertson, Raymond E.; Branthaver, Jan F.; Schabron, John F.

    2012-08-07

    A system for determining parameters and compatibility of a substance such as an asphalt or other petroleum substance uses titration to highly accurately determine one or more flocculation occurrences and is especially applicable to the determination or use of Heithaus parameters and optimal mixing of various asphalt stocks. In a preferred embodiment, automated titration in an oxygen gas exclusive system and further using spectrophotometric analysis (2-8) of solution turbidity is presented. A reversible titration technique enabling in-situ titration measurement of various solution concentrations is also presented.

  3. 3-methyl-2-oxazolidone as a solvent for acid-base titrations.

    PubMed

    Taphorn, J E; Davies, G M; Caruso, J A

    1975-07-01

    3-Methyl-2-oxazolidone has been evaluated as a solvent for the titration of various weak carboxylic acids and phenols. Its high dielectric constant and wide liquid range contribute to its outstanding solvent properties. Tetrabutylammonium hydroxide was the titrant. End-points were determined potentiometrically with a glass-calomel electrode system. FORTRAN computer programs were used to evaluate the results, and relative acid strengths were determined and related to the aqueous acidity. The accuracies obtained are comparable to those for other solvent media. An 800-mV potential span is available in this solvent, which may allow differentiating titrations to be performed. No titrations of acid mixtures were attempted, but it was found possible to distinguish both neutralization steps for salicylic acid and o-phthalic acid.

  4. High-precision gravimetric coulometry using the silver-perchloric acid coulometer: Titration of arsenious oxide with electrogenerated iodine.

    PubMed

    Newton, C M

    1977-06-01

    High-precision gravimetric coulometry with a silver-perchloric acid coulometer is evaluated as an alternative to the conventional titrimetric method. The loss of weight (caused by electrolytic dissolution) of a highly pure silver anode in series with the cathode of a conventional constant-current titration system is measured and related to the number of equivalents of substance titrated. The precision of the method is determined by titrations of the Standard Reference Material 83C arsenious oxide (99.99% pure) with electrogenerated iodine, using biamperometric end-point detection. Depending on the size of the sample, an ultimate precision of 25 ppm is obtained. The assay for 0.5-g samples of the SRM material is 99.993(9) +/- 0.002(5)% purity.

  5. Determination of the degree of substitution of sodium carboxymethylcellulose by potentiometric titration and use of the extended henderson-hasselbalch equation and the simplex method for the evaluation.

    PubMed

    Aggeryd, I; Olin, A

    1985-08-01

    Determination of the degree of substitution, D.S., of purified sodium carboxymethylcellulose, CMC, by potentiometric titration has been studied. Different mathematical descriptions of the acid-base properties of polyelectrolytes were tested on the titration curve. It was found that the extended Henderson-Hasselbalch equation reproduced the measured data well. It was therefore used as a basis for evaluating D.S. by the simplex method. The influence of ionic strength, I, on the shape of the titration curve was investigated and the effect of I on the precision of the titrations was evaluated. The titrations showed a precision of +/- 0.004 D.S.-units and the results were in agreement with those obtained from a dry-ashing method, within +/- 0.02 D.S.-units. The whole method, including preparation of solutions and titration, showed a precision of +/- 0.01 D.S.-units.

  6. Evaluation of the number of ionogenic groups of inulinase by acid-base titration.

    PubMed

    Kovaleva, T A; Holyavka, M G; Rezvan, S G; Kozhedub, S V

    2008-06-01

    Acid base titration showed that Aspergillus awamori inulinase includes 178 asparaginic and glutamic acid residues, 20 histidine, 10 serine, and 34 lysine and tyrosine residues. Denaturation temperature for this enzyme was calculated using analysis of the proportion of stabilizing and destabilizing amino acids in the molecule.

  7. Evaluation of different calculation methods for anti-HBs titration.

    PubMed

    Vranckx, R P

    1989-11-01

    Anti-HBs titre calculations should be standardized to make comparison between various vaccination programmes and between immunogenicity tests in animals more meaningful. For this purpose, we compared four methods of titre calculation in human and animal sera and in the IgM and IgG fractions of these sera. The four calculation methods are parallel line bioassay, Hollinger method, RIA units and regression-calibration curve. From our results, the following can be concluded: The titration curves of anti-HBs/IgM, anti-HBs/IgG and of animal sera do not always run parallel to the WHO standard curve. Thus, the objective quantification of these anti-HBs cannot be obtained with the WHO standard. The Hollinger method and the RIA units method are identical within the range S/P less than 1. They only differ by a factor of 4. Only dilutions giving results in the linear part of the dose-response curve should be considered for titre calculations. The mean of the results for different dilutions should be used to determine the titre. Within the restrictions mentioned, comparable results were obtained with the four methods.

  8. Polarographic indication of the end-point in chelometric titrations with triethylenetetraminehexa-acetic acid.

    PubMed

    Kopanica, M; Stará, V

    1974-10-01

    The application of d.c. and square-wave polarographic measurements for the indication of the end-point in titrations with triethylenetetraminehexa-acetic acid (TTHA) has been studied. TTHA has ten co-ordinating groups and forms complexes with metal to ligand ratios of either 1:1 or 2:1, depending on the metal and experimental conditions. The shape of the titration curves, determined by the dependence of the limiting current (peak current of the square-wave polarographic curve) or of the half-wave potential (peak potential) on the amount of TTHA added indicates the composition of the complex formed in the titration. The composition of this complex as determined from the studied titration curves agrees with that predicted by theory. Titrations with polarographic indication of the end-point were also applied for direct titrations of binary mixtures of metal ions. The resulting titration curves indicated the existence of mixed dinuclear complexes and also the kinetic factors involved in the reactions between two different metal ions and TTHA.

  9. FIA acid-base titrations with a new flow-through pH detector.

    PubMed

    Kahlert, H; Pörksen, J R; Behnert, J; Scholz, F

    2005-08-01

    A pH-sensitive detector for flow-through potentiometry based on a graphite/quinhydrone composite electrode was applied for flow-injection analysis (FIA) titrations. Hydrochloric acid and acetic acid were titrated by injection of samples into a sodium hydroxide carrier solution. System conditions were optimised by variation of flow rate, injection volume and titrant concentration. The parameters sampling frequency, residence time and dispersion coefficient were determined. The evaluation of peak width (time between the two inflection points on each side of the peak), peak area and slope of the raising edge of the peak lead were studied with respect to their use for calibration. Hydrochloric acid and acetic acid could be titrated down to a concentration of 2 x 10(-4) mol L(-1) using 150-microL injection volumes, which is almost ten times lower than can be achieved using colour indicators and a spectrophotometric detection.

  10. Acid-base and chelatometric photo-titrations with photosensors and membrane photosensors.

    PubMed

    Matsuo, T; Masuda, Y; Sekido, E

    1986-08-01

    Photosensors (PS) and membrane photosensors (MPS), which can be immersed in the test solution and facilitate the measurement of concentration, have been developed by miniaturizing an optical system consisting of a light source and a photocell. For use in acid-base or complexometric titrations a poly(vinyl chloride) membrane containing an acid-base or metallochromic indicator can be applied as a coating to the photocell. Spectrophotometric determination of copper(II), and photometric acid-base and chelatometric titrations have been performed with the PS and MPS systems.

  11. Estimating the organic acid contribution to coastal seawater alkalinity by potentiometric titrations in a closed cell.

    PubMed

    Muller, François L L; Bleie, Bjørn

    2008-07-01

    This paper examines the performance of a previously reported, closed cell, potentiometric titration technique [J.M. Hernández-Ayón, S.L. Belli, A. Zirino, Anal. Chim. Acta 394 (1999) 101] for the simultaneous determination of pH, total inorganic carbon (TCO2), total alkalinity (TA), and organic alkalinity (OA) in coastal seawater samples. A novel interpretation of the titration data, as recently proposed by Hernández-Ayón et al. [J.M. Hernández-Ayón, A. Zirino, A.G. Dickson, T. Camiro-Vagas, E. Valenzuela-Espinoza, Limnol. Oceanogr.: Methods 5 (2007) 225] who applied it to waters of unusually high organic matter content, was applied here to fjord surface waters collected over the duration of a phytoplankton bloom. The parameters pH and TCO2--combined with knowledge of boric, phosphate and silicate species concentrations--allowed calculation of all inorganic species that contributed to TA. This inorganic alkalinity term was then subtracted from TA to produce an estimation of OA. Although the OA values obtained were very small (2-22+/-3 micromol L(-1)), they showed a reproducible trend over time in two simultaneous experiments. The organic acids that may have contributed to OA were characterised in back titrations of acidified and CO2-stripped samples with CO2-free NaOH. Two classes of organic titratable species, with pK(a) values around 4.0+/-0.2 and 9.1+/-0.2 were detected. The first occurred in concentrations that co-varied linearly (r2=0.75) with protein-like fluorescence, indicating a marine biological source, but were only weakly correlated (r2=0.46) to OA. By contrast, Class 2 organic species were not significantly correlated to any fluorescence component of either marine or terrestrial origin but were linearly correlated to OA (r2=0.69). These new results reveal that the method proposed by Hernández-Ayón et al. [J.M. Hernández-Ayón, A. Zirino, A.G. Dickson, T. Camiro-Vagas, E. Valenzuela-Espinoza, Limnol. Oceanogr.: Methods 5 (2007) 225] for

  12. Total Acid Value Titration of Hydrotreated Biomass Fast Pyrolysis Oil: Determination of Carboxylic Acids and Phenolics with Multiple End-Point Detection

    SciTech Connect

    Christensen, E.; Alleman, T. L.; McCormick, R. L.

    2013-01-01

    Total acid value titration has long been used to estimate corrosive potential of petroleum crude oil and fuel oil products. The method commonly used for this measurement, ASTM D664, utilizes KOH in isopropanol as the titrant with potentiometric end point determination by pH sensing electrode and Ag/AgCl reference electrode with LiCl electrolyte. A natural application of the D664 method is titration of pyrolysis-derived bio-oil, which is a candidate for refinery upgrading to produce drop in fuels. Determining the total acid value of pyrolysis derived bio-oil has proven challenging and not necessarily amenable to the methodology employed for petroleum products due to the different nature of acids present. We presented an acid value titration for bio-oil products in our previous publication which also utilizes potentiometry using tetrabutylammonium hydroxide in place of KOH as the titrant and tetraethylammonium bromide in place of LiCl as the reference electrolyte to improve the detection of these types of acids. This method was shown to detect numerous end points in samples of bio-oil that were not detected by D664. These end points were attributed to carboxylic acids and phenolics based on the results of HPLC and GC-MS studies. Additional work has led to refinement of the method and it has been established that both carboxylic acids and phenolics can be determined accurately. Use of pH buffer calibration to determine half-neutralization potentials of acids in conjunction with the analysis of model compounds has allowed us to conclude that this titration method is suitable for the determination of total acid value of pyrolysis oil and can be used to differentiate and quantify weak acid species. The measurement of phenolics in bio-oil is subject to a relatively high limit of detection, which may limit the utility of titrimetric methodology for characterizing the acidic potential of pyrolysis oil and products.

  13. Potentiometric Measurement of Transition Ranges and Titration Errors for Acid/Base Indicators

    NASA Astrophysics Data System (ADS)

    Flowers, Paul A.

    1997-07-01

    Sophomore analytical chemistry courses typically devote a substantial amount of lecture time to acid/base equilibrium theory, and usually include at least one laboratory project employing potentiometric titrations. In an effort to provide students a laboratory experience that more directly supports their classroom discussions on this important topic, an experiment involving potentiometric measurement of transition ranges and titration errors for common acid/base indicators has been developed. The pH and visually-assessed color of a millimolar strong acid/base system are monitored as a function of added titrant volume, and the resultant data plotted to permit determination of the indicator's transition range and associated titration error. Student response is typically quite positive, and the measured quantities correlate reasonably well to literature values.

  14. Comparing the Titrations of Mixed-Acid Solutions Using Dropwise and Constant-Flow Techniques

    NASA Astrophysics Data System (ADS)

    Charlesworth, Paul; Seguin, Matthew J.; Chesney, David J.

    2003-11-01

    A mixed-acid solution containing hydrochloric and phosphoric acids was used to determine the error associated with performing a real-time titration. The results were compared against those obtained by performing the titration in a more traditional dropwise addition of titrant near the equivalence points. It was found that the real-time techniques resulted in significantly decreased analysis times while maintaining a low experimental error. The constant-flow techniques were implemented into two different levels of chemistry. It was found that students could successfully utilize the modified experiments. Problems associated with the techniques, major sources of error, and their solutions are discussed. In both cases, the use of the constant-flow setup has increased student recollection of key concepts, such as pKa determination, proper indicator choice, and recognizing the shape of specific titration curves by increasing student interest in the experiment.

  15. Direct sensing of total acidity by chronopotentiometric flash titrations at polymer membrane ion-selective electrodes.

    PubMed

    Gemene, Kebede L; Bakker, Eric

    2008-05-15

    Polymer membrane ion-selective electrodes containing lipophilic ionophores are traditionally interrogated by zero current potentiometry, which, ideally, gives information on the sample activity of ionic species. It is shown here that a discrete cathodic current pulse across an H (+)-selective polymeric membrane doped with the ionophore ETH 5294 may be used for the chronopotentiometric detection of pH in well-buffered samples. However, a reduction in the buffer capacity leads to large deviations from the expected Nernstian response slope. This is explained by the local depletion of hydrogen ions at the sample-membrane interface as a result of the galvanostatically imposed ion flux in direction of the membrane. This depletion is found to be a function of the total acidity of the sample and can be directly monitored chronopotentiometrically in a flash titration experiment. The subsequent application of a baseline potential pulse reverses the extraction process of the current pulse, allowing one to interrogate the sample with minimal perturbation. In one protocol, total acidity is found to be proportional to the magnitude of applied current at the flash titration end point. More conveniently, the square root of the flash titration end point time observed at a fixed applied current is a linear function of the total acid concentration. This suggests that it is possible to perform rapid localized pH titrations at ion-selective electrodes without the need for volumetric titrimetry. The technique is explored here for acetic acid, MES and citric acid with promising results. Polymeric membrane electrodes based on poly(vinyl chloride) plasticized with o-nitrophenyl octyl ether in a 1:2 mass ratio may be used for the detection of acids of up to ca. 1 mM concentration, with flash titration times on the order of a few seconds. Possible limitations of the technique are discussed, including variations of the acid diffusion coefficients and influence of electrical migration.

  16. 40 CFR Appendix A to Part 425 - Potassium Ferricyanide Titration Method

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... concentrations determined by EPA test procedure 376.1 (see 40 CFR 136.3, Table IB, parameter 66 (49 FR 43234... 40 Protection of Environment 30 2011-07-01 2011-07-01 false Potassium Ferricyanide Titration... Appendix A to Part 425—Potassium Ferricyanide Titration Method Source The potassium ferricyanide...

  17. 40 CFR Appendix A to Part 425 - Potassium Ferricyanide Titration Method

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... concentrations determined by EPA test procedure 376.1 (see 40 CFR 136.3, Table IB, parameter 66 (49 FR 43234... 40 Protection of Environment 31 2012-07-01 2012-07-01 false Potassium Ferricyanide Titration..., App. A Appendix A to Part 425—Potassium Ferricyanide Titration Method Source The...

  18. 40 CFR Appendix A to Part 425 - Potassium Ferricyanide Titration Method

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... concentrations determined by EPA test procedure 376.1 (see 40 CFR 136.3, Table IB, parameter 66 (49 FR 43234... 40 Protection of Environment 31 2013-07-01 2013-07-01 false Potassium Ferricyanide Titration..., App. A Appendix A to Part 425—Potassium Ferricyanide Titration Method Source The...

  19. 40 CFR Appendix A to Part 425 - Potassium Ferricyanide Titration Method

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... concentrations determined by EPA test procedure 376.1 (see 40 CFR 136.3, Table IB, parameter 66 (49 FR 43234... 40 Protection of Environment 30 2014-07-01 2014-07-01 false Potassium Ferricyanide Titration..., App. A Appendix A to Part 425—Potassium Ferricyanide Titration Method Source The...

  20. 40 CFR Appendix A to Part 425 - Potassium Ferricyanide Titration Method

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... concentrations determined by EPA test procedure 376.1 (see 40 CFR 136.3, Table IB, parameter 66 (49 FR 43234... 40 Protection of Environment 29 2010-07-01 2010-07-01 false Potassium Ferricyanide Titration... Appendix A to Part 425—Potassium Ferricyanide Titration Method Source The potassium ferricyanide...

  1. [Sensitivity of methods of titration of the vaccine strain of porcine fever virus].

    PubMed

    Koritskaia, M A; Demkina, M M; Vlasova, A N

    2005-01-01

    Methods of titration of the CS vaccine strain of classical swine fever virus were compared in vitro and vivo. The titration in the TL and PK-15 cell culture without cytopathic effect is based on the detection of virus antigen by labeled antibodies. The infection intensity in the cell culture virtually correlated with the antigenic and immunogenic activity of dry vaccine used for swine.

  2. Simple modification of karl-Fischer titration method for determination of water content in colored samples.

    PubMed

    Tavčar, Eva; Turk, Erika; Kreft, Samo

    2012-01-01

    The most commonly used technique for water content determination is Karl-Fischer titration with electrometric detection, requiring specialized equipment. When appropriate equipment is not available, the method can be performed through visual detection of a titration endpoint, which does not enable an analysis of colored samples. Here, we developed a method with spectrophotometric detection of a titration endpoint, appropriate for moisture determination of colored samples. The reaction takes place in a sealed 4 ml cuvette. Detection is performed at 520 nm. Titration endpoint is determined from the graph of absorbance plotted against titration volume. The method has appropriate reproducibility (RSD = 4.3%), accuracy, and linearity (R(2) = 0.997).

  3. Simple Modification of Karl-Fischer Titration Method for Determination of Water Content in Colored Samples

    PubMed Central

    Tavčar, Eva; Turk, Erika; Kreft, Samo

    2012-01-01

    The most commonly used technique for water content determination is Karl-Fischer titration with electrometric detection, requiring specialized equipment. When appropriate equipment is not available, the method can be performed through visual detection of a titration endpoint, which does not enable an analysis of colored samples. Here, we developed a method with spectrophotometric detection of a titration endpoint, appropriate for moisture determination of colored samples. The reaction takes place in a sealed 4 ml cuvette. Detection is performed at 520 nm. Titration endpoint is determined from the graph of absorbance plotted against titration volume. The method has appropriate reproducibility (RSD = 4.3%), accuracy, and linearity (R2 = 0.997). PMID:22567558

  4. Characterization of acid functional groups of carbon dots by nonlinear regression data fitting of potentiometric titration curves

    NASA Astrophysics Data System (ADS)

    Alves, Larissa A.; de Castro, Arthur H.; de Mendonça, Fernanda G.; de Mesquita, João P.

    2016-05-01

    The oxygenated functional groups present on the surface of carbon dots with an average size of 2.7 ± 0.5 nm were characterized by a variety of techniques. In particular, we discussed the fit data of potentiometric titration curves using a nonlinear regression method based on the Levenberg-Marquardt algorithm. The results obtained by statistical treatment of the titration curve data showed that the best fit was obtained considering the presence of five Brønsted-Lowry acids on the surface of the carbon dots with constant ionization characteristics of carboxylic acids, cyclic ester, phenolic and pyrone-like groups. The total number of oxygenated acid groups obtained was 5 mmol g-1, with approximately 65% (∼2.9 mmol g-1) originating from groups with pKa < 6. The methodology showed good reproducibility and stability with standard deviations below 5%. The nature of the groups was independent of small variations in experimental conditions, i.e. the mass of carbon dots titrated and initial concentration of HCl solution. Finally, we believe that the methodology used here, together with other characterization techniques, is a simple, fast and powerful tool to characterize the complex acid-base properties of these so interesting and intriguing nanoparticles.

  5. Titration of Monoprotic Acids with Sodium Hydroxide Contaminated by Sodium Carbonate.

    ERIC Educational Resources Information Center

    Michalowski, Tadeusz

    1988-01-01

    Discusses the effects of using carbon dioxide contaminated sodium hydroxide solution as a titrant for a solution of a weak monoprotic acid and the resulting distortion of the titration curve in comparison to one obtained when an uncontaminated titrant is used. (CW)

  6. Improved methylene blue two-phase titration method for determining cationic surfactant concentration in high-salinity brine.

    PubMed

    Cui, Leyu; Puerto, Maura; López-Salinas, José L; Biswal, Sibani L; Hirasaki, George J

    2014-11-18

    The methylene blue (MB) two-phase titration method is a rapid and efficient method for determining the concentrations of anionic surfactants. The point at which the aqueous and chloroform phases appear equally blue is called Epton's end point. However, many inorganic anions, e.g., Cl(-), NO3(-), Br(-), and I(-), can form ion pairs with MB(+) and interfere with Epton's end point, resulting in the failure of the MB two-phase titration in high-salinity brine. Here we present a method to extend the MB two-phase titration method for determining the concentration of various cationic surfactants in both deionized water and high-salinity brine (22% total dissolved solid). A colorless end point, at which the blue color is completely transferred from the aqueous phase to the chloroform phase, is proposed as titration end point. Light absorbance at the characteristic wavelength of MB is measured using a spectrophotometer. When the absorbance falls below a threshold value of 0.04, the aqueous phase is considered colorless, indicating that the end point has been reached. By using this improved method, the overall error for the titration of a permanent cationic surfactant, e.g., dodecyltrimethylammonium bromide, in deionized (DI) water and high-salinity brine is 1.274% and 1.322% with limits of detection (LOD) of 0.149 and 0.215 mM, respectively. Compared to the traditional acid-base titration method, the error of this improved method for a switchable cationic surfactant, e.g., tertiary amine surfactant (Ethomeen C12), is 2.22% in DI water and 0.106% with LOD of 0.369 and 0.439 mM, respectively.

  7. An automatic system for acidity determination based on sequential injection titration and the monosegmented flow approach.

    PubMed

    Kozak, Joanna; Wójtowicz, Marzena; Gawenda, Nadzieja; Kościelniak, Paweł

    2011-06-15

    An automatic sequential injection system, combining monosegmented flow analysis, sequential injection analysis and sequential injection titration is proposed for acidity determination. The system enables controllable sample dilution and generation of standards of required concentration in a monosegmented sequential injection manner, sequential injection titration of the prepared solutions, data collecting, and handling. It has been tested on spectrophotometric determination of acetic, citric and phosphoric acids with sodium hydroxide used as a titrant and phenolphthalein or thymolphthalein (in the case of phosphoric acid determination) as indicators. Accuracy better than |4.4|% (RE) and repeatability better than 2.9% (RSD) have been obtained. It has been applied to the determination of total acidity in vinegars and various soft drinks. The system provides low sample (less than 0.3 mL) consumption. On average, analysis of a sample takes several minutes. PMID:21641455

  8. Suspension culture titration: A simple method for measuring baculovirus titers.

    PubMed

    Matindoost, Leila; Chan, Leslie C L; Qi, Ying Mei; Nielsen, Lars K; Reid, Steven

    2012-08-01

    The baculovirus-insect cell expression system is an important technology for the production of recombinant proteins and baculovirus-based biopesticides. Budded virus titration is critical when scaling up baculovirus production processes in suspension cultures, to ensure reproducible infections, especially when a low multiplicity of infection (MOI) is applied. In this study, a simple suspension culture titration (SCT) assay was developed that involves accurate measurements of the initial cell densities (ICDs) and peak cell densities (PCDs) of an infected culture, from which the MOI and hence the virus inoculum infectious titer can be estimated, using the established Power-Nielsen baculovirus infection model. The SCT assay was assessed in parallel with two adherent culture-based assays (MTT and AlamarBlue) for the Heliothine baculovirus HaSNPV, and was shown to be more objective, time-efficient and reproducible. The model predicted a linear correlation between log(PCD/ICD) and log(MOI), hence an alternative model-independent SCT assay was also developed, which relies on a well-replicated standard curve relating suspension culture-derived PCD/ICD ratios with plaque or endpoint assay-derived MOIs. Standard curves with excellent linearity were generated for HaSNPV and the industrially significant rAcMNPV, demonstrating the feasibility of this simple titration approach, especially in terms of its applicability to a wide range of virus infection kinetics.

  9. A novel method for the determination of the degree of deacetylation of chitosan by coulometric titration.

    PubMed

    Wang, Chao; Yuan, Fang; Pan, Jiabao; Jiao, Shining; Jin, Ling; Cai, Hongwei

    2014-09-01

    A novel method to determine the degree of deacetylation of chitosan is described. In this method, the coulometric titrant OH- is generated by the electrolysis of water. The OH- reacted with the residual hydrochloric acid in chitosan solution and the degree of deacetylation was obtained by Faraday's law. The optimized experimental parameters in this study were 1.0 mol/L KCl as supporting electrolyte, 15.00 mA as the intensity of constant current, composite glass electrode as indicating electrode couples, double platinum generated electrode-platinum wire auxiliary electrode as working electrode pairs, pH 3.80 as the titration end-point. The degree of deacetylation in the four samples, which varied from 70 to 95% were measured. The results were similar to those from 1H NMR and the standard deviations were lower than 0.5%. With merit of simplicity, convenience, quickness, high accuracy and precision, automatic detection of titration end-point and low-cost, the proposed method will be very useful in the industrial production. PMID:25020083

  10. Acid-base titrations using microfluidic paper-based analytical devices.

    PubMed

    Karita, Shingo; Kaneta, Takashi

    2014-12-16

    Rapid and simple acid-base titration was accomplished using a novel microfluidic paper-based analytical device (μPAD). The μPAD was fabricated by wax printing and consisted of ten reservoirs for reaction and detection. The reaction reservoirs contained various amounts of a primary standard substance, potassium hydrogen phthalate (KHPth), whereas a constant amount of phenolphthalein was added to all the detection reservoirs. A sample solution containing NaOH was dropped onto the center of the μPAD and was allowed to spread to the reaction reservoirs where the KHPth neutralized it. When the amount of NaOH exceeded that of the KHPth in the reaction reservoirs, unneutralized hydroxide ion penetrated the detection reservoirs, resulting in a color reaction from the phenolphthalein. Therefore, the number of the detection reservoirs with no color change determined the concentration of the NaOH in the sample solution. The titration was completed within 1 min by visually determining the end point, which required neither instrumentation nor software. The volumes of the KHPth and phenolphthalein solutions added to the corresponding reservoirs were optimized to obtain reproducible and accurate results for the concentration of NaOH. The μPADs determined the concentration of NaOH at orders of magnitude ranging from 0.01 to 1 M. An acid sample, HCl, was also determined using Na2CO3 as a primary standard substance instead of KHPth. Furthermore, the μPAD was applicable to the titrations of nitric acid, sulfuric acid, acetic acid, and ammonia solutions. The μPADs were stable for more than 1 month when stored in darkness at room temperature, although this was reduced to only 5 days under daylight conditions. The analysis of acidic hot spring water was also demonstrated in the field using the μPAD, and the results agreed well with those obtained by classic acid-base titration.

  11. Comparison of methods for accurate end-point detection of potentiometric titrations

    NASA Astrophysics Data System (ADS)

    Villela, R. L. A.; Borges, P. P.; Vyskočil, L.

    2015-01-01

    Detection of the end point in potentiometric titrations has wide application on experiments that demand very low measurement uncertainties mainly for certifying reference materials. Simulations of experimental coulometric titration data and consequential error analysis of the end-point values were conducted using a programming code. These simulations revealed that the Levenberg-Marquardt method is in general more accurate than the traditional second derivative technique used currently as end-point detection for potentiometric titrations. Performance of the methods will be compared and presented in this paper.

  12. A numerical method of finding potentiometric titration end-points by use of approximative spline functions.

    PubMed

    Ren, K

    1990-07-01

    A new numerical method of determining potentiometric titration end-points is presented. It consists in calculating the coefficients of approximative spline functions describing the experimental data (e.m.f., volume of titrant added). The end-point (the inflection point of the curve) is determined by calculating zero points of the second derivative of the approximative spline function. This spline function, unlike rational spline functions, is free from oscillations and its course is largely independent of random errors in e.m.f. measurements. The proposed method is useful for direct analysis of titration data and especially as a basis for construction of microcomputer-controlled automatic titrators. PMID:18964999

  13. Spectroscopic study on variations in illite surface properties after acid-base titration.

    PubMed

    Liu, Wen-xin; Coveney, R M; Tang, Hong-xiao

    2003-07-01

    FT-IR, Raman microscopy, XRD, 29Si and 27Al MAS NMR, were used to investigate changes in surface properties of a natural illite sample after acid-base potentiometric titration. The characteristic XRD lines indicated the presence of surface Al-Si complexes, preferable to Al(OH)3 precipitates. In the microscopic Raman spectra, the vibration peaks of Si-O and Al-O bonds diminished as a result of treatment with acid, then increased after hydroxide back titration. The varied ratio of signal intensity between (IV)Al and (VI)Al species in 27Al MAS NMR spectra, together with the stable BET surface area after acidimetric titration, suggested that edge faces and basal planes in the layer structure of illite participated in dissolution of structural components. The combined spectroscopic evidence demonstrated that the reactions between illite surfaces and acid-leaching silicic acid and aluminum ions should be considered in the model description of surface acid-base properties of the aqueous illite.

  14. Human saliva and taste responses to acids varying in anions, titratable acidity, and pH.

    PubMed

    Norris, M B; Noble, A C; Pangborn, R M

    1984-02-01

    Twenty subjects recorded perceived sourness of solutions of citric + fumaric and of citric + tartaric acids, at pH 3.5 and titratable acidity (TiA) of 4.0 g/l on a moving chart, while parotid saliva flow was recorded via a sialometer . Sourness intensity and flow were greater when citric was the minor acid than when it was dominant. Subjects varied widely in calculated volume of saliva reservoir, but not flow rate (time to 2/3 reservoir vol.). In tartaric-fumaric acid mixtures varying in pH (3.0-3.75) at a constant TiA of 4.0 g/l, and varying in TiA (3.7-4.6 g/l) at a constant pH of 3.5, sourness intensity and parotid flow increased with acidity and decreased with pH. However, eight subjects with a high flow (HF = 1.2 +/- 0.28 g/2 min) and nine subjects with a low flow (LF = 0.43 +/- 0.11 g/2 min) differed widely: (a) In response to variation in stimulus pH and TiA, HF demonstrated marked alteration in flow, but little change in sourness ; LF responded at a lower absolute level, but showed marked changes in sourness and little change in flow; (b) Salivary pH was higher and Na+ was three times greater for the HF than for the LF subjects; and (c) Salivary Ca++ showed a direct relationship with flow and pH among the HF, but an inverse relationship for the LF subjects.

  15. Hydrogen-ion titrations of amino acids and proteins in solutions containing concentrated electrolyte

    SciTech Connect

    Fergg, F.; Kuehner, D.E.; Blanch, H.W.; Prausnitz, J.M.

    1994-12-01

    This report describes a first attempt to quantify the net charge as a function of solution pH for lysozyme and {alpha}-chymotrypsin at 0.1 M, 1.0 M and 3.0 M ionic strength, (IS). The calculations are based on the residue (titratable group) pK{sub a}`s in the amino-acid sequence of the protein. To determine these pK{sub a}`s, a simple theory was used which assumes that the pK{sub a}`s are independent from each other in the protein and are equal to their pK{sub a} values in free amino-acid solution (Independent-Site Theory, IST). Residue pK{sub a}`s were obtained from amino-acid hydrogen-ion titrations at three different KCl concentrations corresponding to 0.1M, 1.0M and 3.0M ionic strength. After construction of a suitable apparatus, the experimental procedure and data reduction were computerized to perform a large number of titrations. Most measured pK{sub a}`s showed high reproducibility (the difference of pK{sub a} values observed between two experiments was less than 0.05). For IS = 0.1M, observed pK{sub a}`s agreed with literature values to within a few hundredths of a pH unit. Furthermore, the ionic-strength dependence of the pK{sub a}`s followed the trends reported in the literature, viz. pK{sub a} values decrease with increasing ionic strength until they reach a minimum at about IS = 0.5M. At still higher IS, pK{sub a}`s increase as the ionic strength rises to 3M. The known pK{sub a}`s of all titratable groups in a protein were used with the IST to give a first approximation of how the protein net charge varies with pH at high ionic strength. A comparison of the titration curves based on the IST with experimental lysozyme and {alpha}-chymotrypsin titration data indicates acceptable agreement at IS = 0.1M. However, comparison of measured and calculated titration curves at IS = 1M and IS = 3M indicates only quantitative agreement.

  16. A new oxidimetric reagent: potassium dichromate in a strong phosphoric acid medium-VI Potentiometric titration of vanadium(III) alone and in mixture with vanadium(IV).

    PubMed

    Rao, G G; Rao, P K

    1966-09-01

    Vanadium(III) can be titrated at room temperature with potassium dichromate in an 8-12M phosphoric acid medium. Two potential breaks are observed in 12M phosphoric add with 0.2N potassium dichromate, the first corresponding to the oxidation of vanadium(III) to vanadium(IV) and the second to the oxidation of vanadium(IV) to vanadium(V). In titrations with 0.05N dichromate only the first break in potential is clearly observed. The method has been extended to the titration of mixtures of vanadium(III) and vanadium(IV). Conditions have also been found for the visual titration of vanadium(III) using ferroln or barium diphenylamine sulphonate as indicator.

  17. Rapid determination of the equivalence volume in potentiometric acid-base titrations to a preset pH-II Standardizing a solution of a strong base, graphic location of equivalence volume, determination of stability constants of acids and titration of a mixture of two weak acids.

    PubMed

    Ivaska, A

    1974-06-01

    A newly proposed method of titrating weak acids with strong bases is applied to standardize a solution of a strong base, to graphic determination of equivalence volume of acetic acid with an error of 0.2%, to calculate the stability constants of hydroxylammonium ion, boric acid and hydrogen ascorbate ion and to analyse a mixture of acetic acid and ammonium ion with an error of 0.2-0.7%.

  18. Methods for growing and titrating African swine fever virus: field and laboratory samples.

    PubMed

    Carrascosa, Angel L; Bustos, M Jose; de Leon, Patricia

    2011-12-01

    Growing African swine fever virus (ASFV) isolates obtained mainly from the field, but also engineered in the laboratory, is a critical step for diagnosis, titration, or virus infection studies. This unit describes a set of methods and protocols to produce and titrate any ASFV strain in cell cultures. The procedures include (1) basic techniques to prepare virus-sensitive target cells; (2) strategies for growth, concentration, and purification of virus stocks; and (3) the semi-quantitative (end dilution) and quantitative (plaque) assays for the determination of viral titers, and the use of different ASFV-sensitive cells as targets for virus production and titration.

  19. Voltammetric iodometric titration of ascorbic acid with dead-stop end-point detection in fresh vegetables and fruit samples.

    PubMed

    Verdini, R A; Lagier, C M

    2000-07-01

    The present work describes a method for determining ascorbic acid, which combines iodometry with a voltammetric technique to detect the end point of the titration. In addition, the validity of the method applied to natural vegetable or fruit samples was assessed. The results were compared with those obtained by an accurate method such as HPLC using UV detection. Similar values of ascorbic acid for different natural samples were obtained by means of this approach (p > 0.05). The limit of quantification was 0.1 mg. This technique presents the advantage of other electroanalytical methods such as avoiding filtration or ultracentrifugation steps, with the additional benefit of using the platinum electrodes, which are routinely used in the laboratory. These facts allow a rapid and efficient quantification of ascorbic acid with very low cost of reagents and equipment.

  20. Influence of kinetics on the determination of the surface reactivity of oxide suspensions by acid-base titration.

    PubMed

    Duc, M; Adekola, F; Lefèvre, G; Fédoroff, M

    2006-11-01

    The effect of acid-base titration protocol and speed on pH measurement and surface charge calculation was studied on suspensions of gamma-alumina, hematite, goethite, and silica, whose size and porosity have been well characterized. The titration protocol has an important effect on surface charge calculation as well as on acid-base constants obtained by fitting of the titration curves. Variations of pH versus time after addition of acid or base to the suspension were interpreted as diffusion processes. Resulting apparent diffusion coefficients depend on the nature of the oxide and on its porosity.

  1. AMPHOTERIC BEHAVIOR OF COMPLEX SYSTEMS : II. TITRATION OF SULFANILIC ACID-GLYCINE MIXTURES.

    PubMed

    Stearn, A E

    1926-11-20

    Electrometric titrations of glycine, sulfanilic acid, and various mixtures of the two have been made. These mixtures are shown to give a curve which, between their respective isoelectric points, is different from that of either substance. These mixtures have a maximum buffering power at a pH which can be theoretically calculated, and which has the characteristics of an "isoelectric point of the system." Other pairs of ampholytes are shown to act in an analogous manner.

  2. Acid-base titration of melanocortin peptides: evidence of Trp rotational conformers interconversion.

    PubMed

    Fernandez, Roberto M; Vieira, Renata F F; Nakaie, Clóvis R; Lamy, M Teresa; Ito, Amando S

    2005-01-01

    Tryptophantime-resolved fluorescence was used to monitor acid-base titration properties of alpha-melanocyte stimulating hormone (alpha-MSH) and the biologically more potent analog [Nle4, D-Phe7]alpha -MSH (NDP-MSH), labeled or not with the paramagnetic amino acid probe 2,2,6,6-tetramthylpiperidine-N-oxyl-4-amino-4-carboxylic acid (Toac). Global analysis of fluorescence decay profiles measured in the pH range between 2.0 and 11.0 showed that, for each peptide, the data could be well fitted to three lifetimes whose values remained constant. The less populated short lifetime component changed little with pH and was ascribed to Trp g+ chi1 rotamer, in which electron transfer deactivation predominates over fluorescence. The long and intermediate lifetime preexponential factors interconverted along that pH interval and the result was interpreted as due to interconversion between Trp g- and trans chi1 rotamers, driven by conformational changes promoted by modifications in the ionization state of side-chain residues. The differences in the extent of interconversion in alpha-MSH and NDP-MSH are indicative of structural differences between the peptides, while titration curves suggest structural similarities between each peptide and its Toac-labeled species, in aqueous solution. Though less sensitive than fluorescence, the Toac electron spin resonance (ESR) isotropic hyperfine splitting parameter can also monitor the titration of side-chain residues located relatively far from the probe.

  3. Reduction of sample volume and waste generation in acid/base titrations using microelectrodes

    SciTech Connect

    Ekechukwu, A.A.

    1996-03-22

    The Analytical Development Section (ADS) has developed microelectrode methods for use with pH titrations and pH determinations. These microelectrode methods offer increased sensitivity and enable analyses to be done with smaller sample and buffer volumes than are used with standard size electrodes. This report establishes the technical validity of the methods and describes the application of these methods to decreased detection limits, decreased waste generation, and decreased radiation exposure.

  4. Method for improving accuracy of virus titration: standardization of plaque assay for Junin virus.

    PubMed

    Bushar, G; Sagripanti, J L

    1990-10-01

    Titrating infective virus is one of the most important and common techniques in virology. However, after many years of widespread use, the parameters governing the accuracy of titration values are still not well understood. It was found that under conditions currently used for virus titration, only a small percentage of virus in the inoculum is adsorbed onto the cells and thereby detected in the titration assay. The objective of our work was to establish the conditions for a plaque assay which could estimate more accurately the titer of Junin virus. Two different stain methods were compared and several parameters governing plaque formation were studied. The volume of the inoculum appeared as the most important factor affecting observed titer. A linear relationship between the volume of inoculum and the reciprocal apparent titer allowed us to estimate an absolute titer by extrapolation. The approach described here is likely to be applicable to the more accurate estimation of the titer of a wide range of virus.

  5. Influence of chronic respiratory acid-base disorders on acute CO2 titration curve.

    PubMed

    Adrogué, H J; Madias, N E

    1985-04-01

    We have recently shown that background presence of chronic metabolic acid-base disorder markedly alters in vivo acute CO2 titration curve. These studies were carried out to assess the influence of chronic respiratory acid-base disorders on response to acute hypercapnia and to explore whether the chronic level of plasma pH is the factor responsible for alterations in the CO2 titration curve. We compared whole-body responses to acute hypercapnia of dogs with preexisting chronic respiratory alkalosis (n = 8) with that of normal animals (n = 4) and animals with chronic respiratory acidosis (n = 13). Chronic respiratory alkalosis and acidosis, as well as the acute CO2 titrations, were produced in unanesthetized dogs within a large environmental chamber. For comparison with our data on chronic metabolic acidosis and alkalosis, plasma bicarbonate levels, which are secondarily altered in chronic respiratory acid-base disorders, were used as an index of chronic acid-base status of the animals. Results indicate that, as with chronic metabolic acid-base disorders, a larger increment in plasma bicarbonate occurs during acute hypercapnia when steady-state plasma bicarbonate is low (respiratory alkalosis) than when it is high (respiratory acidosis). Yet, in further analogy with the metabolic studies, plasma hydrogen ion concentration is better defended at higher plasma bicarbonate levels in accordance with mathematical relationships defined by the Henderson-Hasselbalch equation. Combined results demonstrate that the influence of chronic acid-base status on whole-body response to acute hypercapnia is independent of initial plasma pH.(ABSTRACT TRUNCATED AT 250 WORDS)

  6. Prediction of aluminum, uranium, and co-contaminants precipitation and adsorption during titration of acidic sediments.

    PubMed

    Tang, Guoping; Luo, Wensui; Watson, David B; Brooks, Scott C; Gu, Baohua

    2013-06-01

    Batch and column recirculation titration tests were performed with contaminated acidic sediments. A generic geochemical model was developed combining precipitation, cation exchange, and surface complexation reactions to describe the observed pH and metal ion concentrations in experiments with or without the presence of CO2. Experimental results showed a slow pH increase due to strong buffering by Al hydrolysis and precipitation and CO2 uptake. The cation concentrations generally decreased at higher pH than those observed in previous tests without CO2. Using amorphous Al(OH)3 and basaluminite precipitation reactions and a cation exchange selectivity coefficient K(Na\\Al) of 0.3, the model approximately described the observed (1) pH titration curve, (2) Ca, Mg, and Mn concentration by cation exchange, and (3) U concentrations by surface complexation with Fe hydroxides at pH < 5 and with liebigite (Ca2UO2(CO3)3·10H2O) precipitation at pH > 5. The model indicated that the formation of aqueous carbonate complexes and competition with carbonate for surface sites could inhibit U and Ni adsorption and precipitation. Our results suggested that the uncertainty in basaluminite solubility is an important source of prediction uncertainty and ignoring labile solid phase Al underestimates the base requirement in titration of acidic sediments.

  7. Prediction of aluminum, uranium, and co-contaminants precipitation and adsorption during titration of acidic sediments.

    PubMed

    Tang, Guoping; Luo, Wensui; Watson, David B; Brooks, Scott C; Gu, Baohua

    2013-06-01

    Batch and column recirculation titration tests were performed with contaminated acidic sediments. A generic geochemical model was developed combining precipitation, cation exchange, and surface complexation reactions to describe the observed pH and metal ion concentrations in experiments with or without the presence of CO2. Experimental results showed a slow pH increase due to strong buffering by Al hydrolysis and precipitation and CO2 uptake. The cation concentrations generally decreased at higher pH than those observed in previous tests without CO2. Using amorphous Al(OH)3 and basaluminite precipitation reactions and a cation exchange selectivity coefficient K(Na\\Al) of 0.3, the model approximately described the observed (1) pH titration curve, (2) Ca, Mg, and Mn concentration by cation exchange, and (3) U concentrations by surface complexation with Fe hydroxides at pH < 5 and with liebigite (Ca2UO2(CO3)3·10H2O) precipitation at pH > 5. The model indicated that the formation of aqueous carbonate complexes and competition with carbonate for surface sites could inhibit U and Ni adsorption and precipitation. Our results suggested that the uncertainty in basaluminite solubility is an important source of prediction uncertainty and ignoring labile solid phase Al underestimates the base requirement in titration of acidic sediments. PMID:23641798

  8. A combination of synchronous fluorescence spectroscopy with chemometric treatment and internal standards in non-aqueous potentiometric titrations of fulvic acids.

    PubMed

    Esteves da Silva, J C; Machado, A A

    1994-12-01

    The acid properties of a soil fulvic acid (sfua) were characterized by potentiometric titration with tetrabutylammonium hydroxide in two non-aqueous solvents with high acid-base resolution power N,N-dimethylformamide (DMF) and acetonitrile. Synchronous fluorescence spectroscopy (SyF) was also used to monitor directly the sfua status during the potentiometric titration in DMF. The potentiometric titration curves showed no clear end-point and the analysis of the sets of spectra obtained at increasing neutralization degree, with a self-modeling curve resolution method (SIMPLISMA), revealed the existence of two components with featureless concentration profiles. Internal standards (maleic, salicylic and p-hydroxylbenzoic acids) were used to determine the amounts of acid groups with different acid strengths in the two non-aqueous solvents. It was shown that the variations observed in the SyF spectra sets of the internal standards are not correlated with those observed in the sfua data. The splitting of the sfua groups in the non-aqueous titration curves seems to be forced artificially depending on the standards used.

  9. Effect of temperature on the acid-base properties of the alumina surface: microcalorimetry and acid-base titration experiments.

    PubMed

    Morel, Jean-Pierre; Marmier, Nicolas; Hurel, Charlotte; Morel-Desrosiers, Nicole

    2006-06-15

    Sorption reactions on natural or synthetic materials that can attenuate the migration of pollutants in the geosphere could be affected by temperature variations. Nevertheless, most of the theoretical models describing sorption reactions are at 25 degrees C. To check these models at different temperatures, experimental data such as the enthalpies of sorption are thus required. Highly sensitive microcalorimeters can now be used to determine the heat effects accompanying the sorption of radionuclides on oxide-water interfaces, but enthalpies of sorption cannot be extracted from microcalorimetric data without a clear knowledge of the thermodynamics of protonation and deprotonation of the oxide surface. However, the values reported in the literature show large discrepancies and one must conclude that, amazingly, this fundamental problem of proton binding is not yet resolved. We have thus undertaken to measure by titration microcalorimetry the heat effects accompanying proton exchange at the alumina-water interface at 25 degrees C. Based on (i) the surface sites speciation provided by a surface complexation model (built from acid-base titrations at 25 degrees C) and (ii) results of the microcalorimetric experiments, calculations have been made to extract the enthalpic variations associated respectively to first and second deprotonation of the alumina surface. Values obtained are deltaH1 = 80+/-10 kJ mol(-1) and deltaH2 = 5+/-3 kJ mol(-1). In a second step, these enthalpy values were used to calculate the alumina surface acidity constants at 50 degrees C via the van't Hoff equation. Then a theoretical titration curve at 50 degrees C was calculated and compared to the experimental alumina surface titration curve. Good agreement between the predicted acid-base titration curve and the experimental one was observed.

  10. The assessment of acid-base disturbance in man by the use of carbon dioxide titration curves.

    PubMed

    Stoker, J B; Kappagoda, C T; Snow, H M; Linden, R J

    1975-02-01

    1. Carbon dioxide titration curves were determined in vivo in dog and man at various degrees of acute non-respiratory acidaemia and alkalaemia. 2. The slope of the CO2 titration curve (delta log Pco2/delta pH) was found to increase with the severity of the acute non-respiratory alkalaemia the slope (delta log Pco2/delta pH) tended towards unity. 3. A simple scheme based on the CO2 titration curves determined in vivo has been proposed for the assessment of acute acid-base disturbances in man. 4. Carbon dioxide titration curves were also determined in vivo in patients with chronic respiratory and non-respiratory acidaemia and it was found that these curves were not significantly different from those obtained in states of acute acid-base disturbances. It is therefore suggested that the scheme described in this paper is applicable to all acid-base disturbances.

  11. Acid-base properties of the alumina surface: influence of the titration procedures on the microcalorimetric results.

    PubMed

    Morel, Jean-Pierre; Marmier, Nicolas; Hurel, Charlotte; Morel-Desrosiers, Nicole

    2009-10-01

    The enthalpy changes associated with the protonation and deprotonation of an alumina surface have been determined on the basis of microcalorimetry experiments and acid-base potentiometric titrations at 25 degrees C. It has been shown that the results may vary significantly according to the experimental procedure. In order to do so, the potentiometric and microcalorimetric titrations have been carried out first from an acidic pH to basic pH and second from a pH near the pH(zpc) of alumina to acidic or basic pH. It has been demonstrated that the pK(a) values deduced from the potentiometric titrations are the same whatever the experimental protocol whereas the only way to obtain meaningful enthalpies of proton exchange is to carry out microcalorimetric titrations by starting around the point of zero charge.

  12. A nine-point pH titration method to determine low-concentration VFA in municipal wastewater.

    PubMed

    Ai, Hainan; Zhang, Daijun; Lu, Peili; He, Qiang

    2011-01-01

    Characterization of volatile fatty acid (VFA) in wastewater is significant for understanding the wastewater nature and the wastewater treatment process optimization based on the usage of Activated Sludge Models (ASMs). In this study, a nine-point pH titration method was developed for the determination of low-concentration VFA in municipal wastewater. The method was evaluated using synthetic wastewater containing VFA with the concentration of 10-50 mg/l and the possible interfering buffer systems of carbonate, phosphate and ammonium similar to those in real municipal wastewater. In addition, the further evaluation was conducted through the assay of real wastewater using chromatography as reference. The results showed that the recovery of VFA in the synthetic wastewater was 92%-102 and the coefficient of variance (CV) of reduplicate measurements 1.68%-4.72%. The changing content of the buffering substances had little effect on the accuracy of the method. Moreover, the titration method was agreed with chromatography in the determination of VFA in real municipal wastewater with R(2)= 0.9987 and CV =1.3-1.7. The nine-point pH titration method is capable of satisfied determination of low-concentration VFA in municipal wastewater.

  13. Quantitative analysis of citric acid/sodium hypophosphite modified cotton by HPLC and conductometric titration.

    PubMed

    Ye, Tao; Wang, Bijia; Liu, Jian; Chen, Jiangang; Yang, Yiqi

    2015-05-01

    Isocratic HPLC was used in conjunction with conductometric titration to quantitatively examine the modification of cotton cellulose by citric acid (CA)/sodium hypophosphite (SHP). CA/SHP had been extensively used as a green crosslinking agent for enhancement of cellulose and other carbohydrate polymers without in-depth understanding of the mechanisms. The current study investigated all identifiable secondary polycarboxylic acids from CA decomposition in the CA/SHP-cellulose system under various curing conditions. It was found that CA decomposition was more sensitive to temperature compared with the desirable esterification reaction. Two crosslinking mechanisms, namely ester crosslinking and SHP crosslinking were responsible for the observed improvement in crease resistance of CA/SHP treated cotton fabrics. An oligomer of citraconic acid (CCA) and/or itaconic acid (IA) was identified as a possible contributor to fabric yellowing. Finally, the crease resistance of fabrics correlated strongly with CA preservation in polyol-added CA/SHP crosslinking systems. The dosage of polyol should be held below an inflexion point to keep the undesirable competition against cellulose minimum. The combination of HPLC and conductometric titration was demonstrated to be useful in studying the CA/SHP-cellulose crosslinking system. The findings have implications for better application of CA/SHP in polysaccharide modifications in general.

  14. Potentiometric study of reaction between periodate and iodide as their tetrabutylammonium salts in chloroform. Application to the determination of iodide and potentiometric detection of end points in acid-base titrations in chloroform.

    PubMed

    Pournaghi-Azar, M H; Farhadi, K

    1995-03-01

    A potentiometric method for the titration of tetrabutylammonium iodide (TBAI) in chloroform using tetrabutylammonium periodate (TBAPI) as a strong and suitable oxidizing reagent is described. The potentiometric conditions were optimized and the equilibrium constants of the reactions occurring during the titration were determined. The method was used for the determination of iodide both in chloroform and aqueous solutions after extraction into chloroform as ion-association with tetraphenylarsonium. The reaction between TBAPI and TBAI was also used as acid indicator for the potentiometric detection of end points of acid-base titrations in chloroform.

  15. Determination of microscopic acid-base parameters from NMR-pH titrations.

    PubMed

    Szakács, Zoltán; Kraszni, Márta; Noszál, Béla

    2004-03-01

    The theory and practice of proton microspeciation based on NMR-pH titrations are surveyed. Principles of bi-, tri-, tetra-, and n-protic microequilibrium systems are discussed. Evaluation methods are exemplified by case studies on bi- and tetraprotic biomolecules. Selection criteria and properties of 'reporter' NMR nuclei are described. Literature data on complete microspeciations of small ligands and site-specific basicity characterizations of peptides and proteins are critically reviewed.

  16. A novel method for the titration of recombinant virus stocks by ELISPOT assay.

    PubMed

    Cheong, Siew Chiat; Clément, Nathalie; Velu, Thierry; Brandenburger, Annick

    2003-05-01

    The development of vectors for gene therapy requires the definition of quality control parameters such as titration, contamination, transduction efficiency and biological effects in defined model systems. For most viral vectors, the classical titration by plaque formation is not applicable, because vectors are defective for replication and packaging cell lines are not always available. In particular, for vectors derived from the autonomous parvovirus MVM(p), the titration method used currently is based on the amplification of the viral genome inside an infected cell, which can then be revealed with a specific radioactive probe (J. Virol. 63 (1989) 1023). In situ hybridization allows to titrate wild-type virus as well as vectors, using probes that are specific for the substituted viral genes or for the transgene, respectively. This method is, however, time consuming, making the simultaneous titration of large numbers of samples difficult. The use of a radioactive probe requires an adequate facility. An ELISPOT method that allows for rapid titration of up to 23 vector stocks in one 96 well dish was devised. This method is based on the actual expression of the transgene. Compared to in situ hybridization, titers obtained by the ELISPOT method were in general equivalent or higher. However, for some vector stocks the ELISPOT titers were repeatedly lower, indicating that in situ hybridization does not give an accurate measure of transducing units. Our model system is recombinant parvovirus MVM expressing human IL2, but the method should be adaptable to other vectors expressing transgenes that are secreted and for which antibodies are available.

  17. Iodine--iodide reference electrode for potentiometric titrations of bases in acetic acid.

    PubMed

    Maccà, Carlo; Soldà, Lidia

    2002-03-01

    The suitability of the half-cell Pt/I2, KI in glacial acetic acid as reference electrode for potentiometric titrations of weak bases in the same medium was investigated. This electrode is easily prepared. It has been found very convenient in use and performing at least as well as the best electrodes used for the same purpose. Some phenomenological aspects of the behaviour of the above half-cell, of interest for the use as the reference electrode in the laboratory practice have been observed, that show important differences respect aqueous medium and other organic solvents.

  18. New approach in the treatment of data from an acid-base potentiometric titrationI. Monocomponent systems of monofunctional acids and bases.

    PubMed

    Maslarska, Vania; Tencheva, Jasmina; Budevsky, Omortag

    2003-01-01

    Based on precise analysis of the acid-base equilibrium, a new approach in the treatment of experimental data from a potentiometric titration is proposed. A new general formula giving explicitly the relation V=f([H(+)]) is derived, valid for every acid-base titration, which includes mono- and polyfunctional protolytes and their mixtures. The present study is the first practical application of this formula for the simplest case, the analysis of one monofunctional protolyte. The collected mV data during the titration are converted into pH-values by means of an auto pH-calibration procedure, thus avoiding preliminary preparation of the measuring system. The mentioned pH-calibration method is applicable also in water-organic mixtures and allows the quantitative determination of sparingly soluble substances (particularly pharmaceuticals). The treatment of the data is performed by means of ready-to-use software products, which makes the proposed approach accessible for a wide range of applications.

  19. Tested Demonstrations: Comparison of Strong Acid and Weak Acid Titration Curves.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1979-01-01

    A lecture demonstration is presented for comparing titration curves. A plot of pH vs volume of strong base is produced by connecting the external output of a pH meter to a strip recorder. Thus, pH is recorded as a function of time. (BB)

  20. A Universal Method for Fishing Target Proteins from Mixtures of Biomolecules using Isothermal Titration Calorimetry

    SciTech Connect

    Zhou, X.; Sun, Q; Kini, R; Sivaraman, J

    2008-01-01

    The most challenging tasks in biology include the identification of (1) the orphan receptor for a ligand, (2) the ligand for an orphan receptor protein, and (3) the target protein(s) for a given drug or a lead compound that are critical for the pharmacological or side effects. At present, several approaches are available, including cell- or animal-based assays, affinity labeling, solid-phase binding assays, surface plasmon resonance, and nuclear magnetic resonance. Most of these techniques are not easy to apply when the target protein is unknown and the compound is not amenable to labeling, chemical modification, or immobilization. Here we demonstrate a new universal method for fishing orphan target proteins from a complex mixture of biomolecules using isothermal titration calorimetry (ITC) as a tracking tool. We took snake venom, a crude mixture of several hundred proteins/peptides, as a model to demonstrate our proposed ITC method in tracking the isolation and purification of two distinct target proteins, a major component and a minor component. Identities of fished out target proteins were confirmed by amino acid sequencing and inhibition assays. This method has the potential to make a significant advancement in the area of identifying orphan target proteins and inhibitor screening in drug discovery and characterization.

  1. Anhydrous formic acid and acetic anhydride as solvent or additive in nonaqueous titrations.

    PubMed

    Buvári-Barcza, A; Tóth, I; Barcza, L

    2005-09-01

    The use and importance of formic acid and acetic anhydride (Ac2O) is increasing in nonaqueous acid-base titrations, but their interaction with the solutes is poorly understood. This paper attempts to clarify the effect of the solvents; NMR and spectrophotometric investigations were done to reveal the interactions between some bases and the mentioned solvents. Anhydrous formic acid is a typical protogenic solvent but both the relative permittivity and acidity are higher than those of acetic acid (mostly used in assays of bases). These differences originate from the different chemical structures: liquid acetic acid contains basically cyclic dimers while formic acid forms linear associates. Ac2O is obviously not an acidic but an aprotic (very slightly protophilic) solvent, which supposedly dissociates slightly into acetyl (CH3CO+) and acetate (AcO-) ions. In fact, some bases react with Ac2O forming an associate: the Ac+ group is bound to the delta- charged atom of the reactant while AcO- is associated with the delta+ group at appropriate distance.

  2. Acid-base titration of streptococci and the physical states of intracellular ions.

    PubMed

    Marquis, R E; Porterfield, N; Matsumura, P

    1973-05-01

    Acid titrations of intact and butanol-treated cells of Streptococcus faecalis revealed that nearly all of the intracellular K(+) ions could diffuse into the suspending medium in association with small anions, including ribonucleic acid breakdown products, when the cell membrane was damaged. In contrast, nearly all of the intracellular Mg(2+) ions appeared to be firmly bound to stable internal cell components but could be displaced reversibly by hydronium ions. The cell membrane acted as a barrier to ion movements, and Mg(2+) displacement from intact cells required more acid conditions, by as much as 2.5 pH units, than did displacement from butanol-damaged cells. Some 15 to 20% of the cell magnesium appeared to be associated with surface structures in that it could be removed at pH 7 with ethylenediaminetetraacetic acid or displaced by Co(2+), Ni(2+), Sr(2+), or La(3+). Magnesium could be displaced from isolated cell walls and membranes by hydronium ions in the pH range from 5 to 3, over which carboxyl groups were titrated. Displacement of magnesium from ribosomes also took place between pH 5 and 3, but it was more difficult to identify the magnesium-releasing groups because both protein carboxyl groups and purine and pyrimidine ring nitrogens can become protonated in this pH range. Isolated protoplast membranes remained structurally intact when completely depleted of magnesium. Furthermore, protoplasts isolated from intact cells were found to have greatly enhanced resistance to osmotic shock in acid media, even when solute loss was not extensive. Osmotic resistance was lost when the protoplasts were again placed in neutral media, and this reversibility suggested that acidification caused changes in the physical properties of membranes as well as solute leakage from cells.

  3. Simultaneous measurement of gastric acid and duodenal alkali secretion by in situ titration in health and disease.

    PubMed

    Dubey, P; Nundy, S

    1983-12-01

    We have devised a technique for simultaneously measuring the acid secretion into the stomach and alkali into the duodenum by in situ titration using a modification of the technique of Fordtran and Walsh. Using this technique, the results of acid and alkali secretion measured simultaneously were identical with those obtained using the conventional aspiration method on separate days. In response to stimulation with pentagastrin acid output was 17.2 +/- 1.4 vs 15.4 +/- 1.9 mmol/h and alkali response with secretin was 16.0 +/- 0.8 vs 14.4 +/- 1.5 mmol/h. The response to food was measured in 10 control subjects, 10 patients with duodenal ulcer, and 10 patients with pancreatitis. In controls, the acid and alkaline secretion were similar (15.8 +/- 1.7 vs 18.2 +/- 1.3 mmol/h), in patients with duodenal ulcer acid secretion was significantly greater than alkaline secretion (31.9 +/- 2.2 vs 21.9 +/- 1.7 mmol/h), and in patients with pancreatitis the alkali secretion was significantly less than acid (19.8 +/- 1.9 mmol/h acid vs 11.4 +/- 0.6 mmol/h alkali). It can, therefore, be concluded that in response to food the patients with duodenal ulcer are significant hypersecretors of acid (DU acid greater than DU alkali output) and patients with pancreatitis are significant hyposecretors of alkali (pancreatitis-alkaline output less than acid output) and normal subjects secrete equal amounts of acid and alkali.

  4. A novel EBNA-1 titration method and putative anti-EBNA-1 protein.

    PubMed

    Nonoyama, M; Wen, L T; Tabata, T; Tanaka, A

    1988-09-01

    A novel and rapid EBNA-1 titration method has been developed which uses immunoprecipitation of specific DNA-protein complexes with EBNA-1-positive serum. The method is more sensitive than the conventional immunofluorescence method and has potential value as a diagnostic reagent for clinical laboratories. TPA induction of putative anti-EBNA-1 protein of cellular origin is discussed, which may play a key role for the shift from latent to lytic replication of EBV.

  5. Isothermal titration calorimetry - a new method for the quantification of microbial degradation of trace pollutants.

    PubMed

    Mariana, F; Buchholz, F; Harms, H; Yong, Z; Yao, J; Maskow, T

    2010-07-01

    The environmental fate and, in particular, biodegradation rates of hydrophobic organic compounds (HOC) are of high interest due to the ubiquity, persistence, and potential health effects of these compounds. HOC tend to interact with bioreactor materials and sampling devices and are frequently volatile, so that conventionally derived degradation parameters are often biased. We report on the development and validation of a novel calorimetric approach that serves to gain real time information on the kinetics and the physiology of HOC bioconversion in aqueous systems while overcoming weaknesses of conventional biodegradation experiments. Soil bacteria Mycobacterium frederiksbergense LB501T, Rhodococcus erythropolis K2-3 and Pseudomonas putida G7 were exposed to pulsed titrations of dissolved anthracene, 4-(2,4-dichlorophenoxy)butyric acid or naphthalene, respectively, and the thermal responses were monitored. The combinations of strains and pollutants were selected as examples for complete and partial biodegradation and complete degradation with storage product formation, respectively. Heat production signals were interpreted thermodynamically and in terms of Michaelis-Menten kinetics. The half-saturation constant k(D) and the degradation rate r(D)(Max) were derived. Comparison with conventional methods shows the suitability to extract kinetic degradation parameters of organic trace pollutants from simple ITC experiments, while thermodynamic interpretation provided further information about the metabolic fate of HOC compounds.

  6. The active titration method for measuring local hydroxyl radical concentration

    NASA Technical Reports Server (NTRS)

    Sprengnether, Michele; Prinn, Ronald G.

    1994-01-01

    We are developing a method for measuring ambient OH by monitoring its rate of reaction with a chemical species. Our technique involves the local, instantaneous release of a mixture of saturated cyclic hydrocarbons (titrants) and perfluorocarbons (dispersants). These species must not normally be present in ambient air above the part per trillion concentration. We then track the mixture downwind using a real-time portable ECD tracer instrument. We collect air samples in canisters every few minutes for roughly one hour. We then return to the laboratory and analyze our air samples to determine the ratios of the titrant to dispersant concentrations. The trends in these ratios give us the ambient OH concentration from the relation: dlnR/dt = -k(OH). A successful measurement of OH requires that the trends in these ratios be measureable. We must not perturb ambient OH concentrations. The titrant to dispersant ratio must be spatially invariant. Finally, heterogeneous reactions of our titrant and dispersant species must be negligible relative to the titrant reaction with OH. We have conducted laboratory studies of our ability to measure the titrant to dispersant ratios as a function of concentration down to the few part per trillion concentration. We have subsequently used these results in a gaussian puff model to estimate our expected uncertainty in a field measurement of OH. Our results indicate that under a range of atmospheric conditions we expect to be able to measure OH with a sensitivity of 3x10(exp 5) cm(exp -3). In our most optimistic scenarios, we obtain a sensitivity of 1x10(exp 5) cm(exp -3). These sensitivity values reflect our anticipated ability to measure the ratio trends. However, because we are also using a rate constant to obtain our (OH) from this ratio trend, our accuracy cannot be better than that of the rate constant, which we expect to be about 20 percent.

  7. Procedure for the quantitative determination of mixtures of nucleic Acid components based on multivariate spectrophotometric Acid-base titrations.

    PubMed

    Saurina, J; Hernández-Cassou, S; Tauler, R; Izquierdo-Ridorsa, A

    1999-01-01

    A new procedure for the quantitative determination of mixtures of nucleic acid components, based on continuous spectrophotometric acid-base titrations and multivariate curve resolution, is proposed. The procedure simultaneously takes into account the spectroscopic and acid-base properties of the compounds, which leads to a higher selectivity. Furthermore, quantitative determination of an analyte in a complex mixture is performed using a synthetic solution as standard containing only the analyte of interest. An intrinsic difficulty in the analysis of spectrometric titration data is the presence of rank deficiency due to closure for the mixtures of two or more compounds. An additional problem can be encountered in some mixtures if species spectra or species concentration profiles are practically identical (rank overlap). However, even in the presence of these rank difficulties, accurate quantitation with prediction errors lower than 5% was obtained. The presence of unknown and uncalibrated interferences in the samples does not affect the quantitative determination of the analyte of interest. The proposed procedure was successfully applied to the analysis of real samples (pharmaceuticals) using synthetic external standards.

  8. Semi-automatic version of the potentiometric titration method for characterization of uranium compounds.

    PubMed

    Cristiano, Bárbara F G; Delgado, José Ubiratan; da Silva, José Wanderley S; de Barros, Pedro D; de Araújo, Radier M S; Dias, Fábio C; Lopes, Ricardo T

    2012-09-01

    The potentiometric titration method was used for characterization of uranium compounds to be applied in intercomparison programs. The method is applied with traceability assured using a potassium dichromate primary standard. A semi-automatic version was developed to reduce the analysis time and the operator variation. The standard uncertainty in determining the total concentration of uranium was around 0.01%, which is suitable for uranium characterization and compatible with those obtained by manual techniques. PMID:22406220

  9. Canadian Tire Money: An Analogy for Use When Discussing Weak Acid Strong Base Titrations

    NASA Astrophysics Data System (ADS)

    Last, Arthur M.

    2003-12-01

    A simple analogy can often provide an instructor with a means of helping students to understand an unfamiliar concept. An analogy involving money can be particularly helpful as most students have experience in dealing with a range of financial transactions in their everyday lives. In this article, use is made of the practice of one well-known Canadian retail chain in returning to its customers a small percentage of an item's purchase price in the form of imitation bank notes that can subsequently be spent in the chain's stores. An analogy is drawn between this practice and the determination of the pKa of a weak acid by titrating it with a strong base, taking into account the hydrolysis of the anion produced.

  10. FIA titrations of phenothiazine derivatives in aqueous micellar and non-aqueous media.

    PubMed

    Nemcová, Irena; Nesmerák, Karel; Rychlovský, Petr; Koutníková, Jitka

    2005-02-15

    New methods of flow injection analysis (FIA) neutralization titrations of phenothiazine derivatives in aqueous micellar medium of a cationic surfactant using potentiometric and spectrophotometric detection were proposed; titrations with a mixing gradient chamber and high-speed titrations were compared. The FIA titration method in non-aqueous media based on an official method of determination (titration with perchloric acid in anhydrous acetic acid) was also developed. Under optimized reaction conditions and flow-through parameters, the calibration range and equations, the sensitivity, and the repeatability of all methods were found and discussed. All titrations were assayed for medicinal forms.

  11. Equilibrium titrations of acid-induced unfolding-refolding and salt-induced molten globule of cytochrome c by FT-IR spectroscopy.

    PubMed

    Dong, Aichun; Lam, Troy

    2005-04-01

    Despite extensive investigations on the acid-unfolded and acid/salt-induced molten globule(-like) states of cytochrome c using variety of techniques, structural features of the acid-unfolded state in terms of residual secondary structures and the structural transition between the acid-unfolded and acid/salt-refolded states have not been fully characterized beyond the circular dichroism (CD) spectroscopy. It is unusual that secondary structure(s) of the unfolded state leading to the molten globule state, an important protein folding intermediate, as determined by CD was not fully corroborated by independent experimental method(s). In this study, we carried out an equilibrium titration of acid-induced unfolding and subsequent acid- and salt-induced refolding of cytochrome c using Fourier transform infrared spectroscopy. The spectral profiles of the equilibrium titration reveal new structural details about the acid-unfolded state and the structural transition associated with the acid/salt-refolded molten globule(-like) states of cytochrome c.

  12. pH indicator titration: a novel fast pKa determination method.

    PubMed

    Kong, Xiaole; Zhou, Tao; Liu, Zudong; Hider, Robert C

    2007-10-01

    This study describes a fast spectrophotometic titration method for apparent ionization constant (pKa) determination. In this method, a Universal pH indicator is utilized instead of the conventional pH electrode. An autoburette is set to add HCl at a constant rate to a vigorously stirred 1 cm UV cuvette which contains sample and indicator solution. A spectrophotometer continuously records the spectra. Acquired spectral data are processed by calculating the pH from the indicator spectra in the visible region and extracting sample spectra from the UV region. Five compounds possessing pKa values in the range 2-10 were investigated. These results differed from measurements by conventional spectrophotometric titration by +/-0.05 to +/-0.10 log unit.

  13. The pH at the First Equivalence Point in the Titration of a Diprotic Acid

    NASA Astrophysics Data System (ADS)

    Ault, Addison

    2003-12-01

    Some readers will note a similarity between this approach and the one I took in a paper entitled “Do pH in Your Head” (2). In an example in that article the isoelectric pH of glycine (the pH at which the average charge of a glycine molecule is zero), has the value of 6.0, which is exactly half-way between 2.4, the pKa of the carboxyl group of glycine, and 9.6, the pKa of the ammonium group of glycine. This is what one would expect when realizing that a solution of neutral glycine right out of the bottle is equivalent to glycine obtained by titration of the conjugate acid of glycine to the first equivalence point. Those who are interested might want to consider why the isoelectric pH of an “acidic” amino acid, such as alanine, is exactly half-way between the pKa values of the two carboxyl groups, and why the isoelectric pH of a “basic” amino acid such as lysine is exactly half-way between the pKa values of the two ammonium groups.

  14. Geochemical Modeling of Reactions and Partitioning of Trace Metals and Radionuclides during Titration of Contaminated Acidic Sediments

    SciTech Connect

    Zhang, Fan; Parker, Jack C.; Luo, Wensui; Spalding, Brian Patrick; Brooks, Scott C; Watson, David B; Jardine, Philip M; Gu, Baohua

    2008-01-01

    Many geochemical reactions that control aqueous metal concentrations are directly affected by solution pH. However, changes in solution pH are strongly buffered by various aqueous phase and solid phase precipitation/dissolution and adsorption/desorption reactions. The ability to predict acid-base behavior of the soil-solution system is thus critical to predict metal transport under variable pH conditions. This study was undertaken to develop a practical generic geochemical modeling approach to predict aqueous and solid phase concentrations of metals and anions during conditions of acid or base additions. The method of Spalding and Spalding was utilized to model soil buffer capacity and pH-dependent cation exchange capacity by treating aquifer solids as a polyprotic acid. To simulate the dynamic and pH-dependent anion exchange capacity, the aquifer solids were simultaneously treated as a polyprotic base controlled by mineral precipitation/dissolution reactions. An equilibrium reaction model that describes aqueous complexation, precipitation, sorption and soil buffering with pH-dependent ion exchange was developed using HydroGeoChem v5.0 (HGC5). Comparison of model results with experimental titration data of pH, Al, Ca, Mg, Sr, Mn, Ni, Co, and SO{sub 4}{sup 2-} for contaminated sediments indicated close agreement, suggesting that the model could potentially be used to predict the acid-base behavior of the sediment-solution system under variable pH conditions.

  15. Geochemical modeling of reactions and partitioning of trace metals and radionuclides during titration of contaminated acidic sediments.

    PubMed

    Zhang, Fan; Luo, Wensui; Parker, Jack C; Spalding, Brian P; Brooks, Scott C; Watson, David B; Jardine, Philip M; Gu, Baohua

    2008-11-01

    Many geochemical reactions that control aqueous metal concentrations are directly affected by solution pH. However, changes in solution pH are strongly buffered by various aqueous phase and solid phase precipitation/dissolution and adsorption/desorption reactions. The ability to predict acid-base behavior of the soil-solution system is thus critical to predict metal transport under variable pH conditions. This studywas undertaken to develop a practical generic geochemical modeling approach to predict aqueous and solid phase concentrations of metals and anions during conditions of acid or base additions. The method of Spalding and Spalding was utilized to model soil buffer capacity and pH-dependent cation exchange capacity by treating aquifer solids as a polyprotic acid. To simulate the dynamic and pH-dependent anion exchange capacity, the aquifer solids were simultaneously treated as a polyprotic base controlled by mineral precipitation/ dissolution reactions. An equilibrium reaction model that describes aqueous complexation, precipitation, sorption and soil buffering with pH-dependent ion exchange was developed using HydroGeoChem v5.0 (HGC5). Comparison of model results with experimental titration data of pH, Al, Ca, Mg, Sr, Mn, Ni, Co, and SO4(2-) for contaminated sediments indicated close agreement suggesting that the model could potentially be used to predictthe acid-base behavior of the sediment-solution system under variable pH conditions.

  16. Assembling and Using an LED-Based Detector to Monitor Absorbance Changes during Acid-Base Titrations

    ERIC Educational Resources Information Center

    Santos, Willy G.; Cavalheiro, E´der T. G.

    2015-01-01

    A simple photometric assembly based in an LED as a light source and a photodiode as a detector is proposed in order to follow the absorbance changes as a function of the titrant volume added during the course of acid-base titrations in the presence of a suitable visual indicator. The simplicity and low cost of the electronic device allow the…

  17. Kinetic properties of two Rhizopus exo-polygalacturonase enzymes hydrolyzing galacturonic acid oligomers using isothermal titration calorimetry

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The kinetic characteristics of two Rhizopus oryzae exo-polygalacturonases acting on galacturonic acid oligomers (GalpA) were determined using isothermal titration calorimetry (ITC). RPG15 hydrolyzing (GalpA)2 demonstrated a Km of 55 uM and kcat of 10.3 s^-1^ while RPG16 was shown to have greater af...

  18. Flow cytometry as an improved method for the titration of Chlamydiaceae and other intracellular bacteria.

    PubMed

    Käser, T; Pasternak, J A; Hamonic, G; Rieder, M; Lai, K; Delgado-Ortega, M; Gerdts, V; Meurens, F

    2016-05-01

    Chlamydiaceae is a family of intracellular bacteria causing a range of diverse pathological outcomes. The most devastating human diseases are ocular infections with C. trachomatis leading to blindness and genital infections causing pelvic inflammatory disease with long-term sequelae including infertility and chronic pelvic pain. In order to enable the comparison of experiments between laboratories investigating host-chlamydia interactions, the infectious titer has to be determined. Titer determination of chlamydia is most commonly performed via microscopy of host cells infected with a serial dilution of chlamydia. However, other methods including fluorescent ELISpot (Fluorospot) and DNA Chip Scanning Technology have also been proposed to enumerate chlamydia-infected cells. For viruses, flow cytometry has been suggested as a superior alternative to standard titration methods. In this study we compared the use of flow cytometry with microscopy and Fluorospot for the titration of C. suis as a representative of other intracellular bacteria. Titer determination via Fluorospot was unreliable, while titration via microscopy led to a linear read-out range of 16 - 64 dilutions and moderate reproducibility with acceptable standard deviations within and between investigators. In contrast, flow cytometry had a vast linear read-out range of 1,024 dilutions and the lowest standard deviations given a basic training in these methods. In addition, flow cytometry was faster and material costs were lower compared to microscopy. Flow cytometry offers a fast, cheap, precise, and reproducible alternative for the titration of intracellular bacteria like C. suis. © 2016 International Society for Advancement of Cytometry. PMID:26849001

  19. Flow cytometry as an improved method for the titration of Chlamydiaceae and other intracellular bacteria.

    PubMed

    Käser, T; Pasternak, J A; Hamonic, G; Rieder, M; Lai, K; Delgado-Ortega, M; Gerdts, V; Meurens, F

    2016-05-01

    Chlamydiaceae is a family of intracellular bacteria causing a range of diverse pathological outcomes. The most devastating human diseases are ocular infections with C. trachomatis leading to blindness and genital infections causing pelvic inflammatory disease with long-term sequelae including infertility and chronic pelvic pain. In order to enable the comparison of experiments between laboratories investigating host-chlamydia interactions, the infectious titer has to be determined. Titer determination of chlamydia is most commonly performed via microscopy of host cells infected with a serial dilution of chlamydia. However, other methods including fluorescent ELISpot (Fluorospot) and DNA Chip Scanning Technology have also been proposed to enumerate chlamydia-infected cells. For viruses, flow cytometry has been suggested as a superior alternative to standard titration methods. In this study we compared the use of flow cytometry with microscopy and Fluorospot for the titration of C. suis as a representative of other intracellular bacteria. Titer determination via Fluorospot was unreliable, while titration via microscopy led to a linear read-out range of 16 - 64 dilutions and moderate reproducibility with acceptable standard deviations within and between investigators. In contrast, flow cytometry had a vast linear read-out range of 1,024 dilutions and the lowest standard deviations given a basic training in these methods. In addition, flow cytometry was faster and material costs were lower compared to microscopy. Flow cytometry offers a fast, cheap, precise, and reproducible alternative for the titration of intracellular bacteria like C. suis. © 2016 International Society for Advancement of Cytometry.

  20. A rapid method for titrating baculovirus stocks using the Sf-9 Easy Titer cell line.

    PubMed

    Hopkins, Ralph; Esposito, Dominic

    2009-09-01

    A new rapid method for titrating baculovirus stocks has been developed using a novel cell line Sf-9 Easy Titer (Sf-9ET). The Sf-9ET cell line has been transfected with plasmid DNA containing the enhanced green fluorescent protein (eGFP) gene under the control of the baculovirus polyhedrin promoter. When used in the titration assay, the Sf-9ET cells turn green when they are infected with baculovirus due to the activation of the polyhedrin promoter/eGFP complex by baculovirus gene products expressed during the infection. Using a 96-well plate format end-point dilution assay, baculovirus titers can be determined in three days using a fluorescence microscope.

  1. Titration Curves: Fact and Fiction.

    ERIC Educational Resources Information Center

    Chamberlain, John

    1997-01-01

    Discusses ways in which datalogging equipment can enable titration curves to be measured accurately and how computing power can be used to predict the shape of curves. Highlights include sources of error, use of spreadsheets to generate titration curves, titration of a weak acid with a strong alkali, dibasic acids, weak acid and weak base, and…

  2. Standardization of a neutralizing anti-vaccinia antibodies titration method: an essential step for titration of vaccinia immunoglobulins and smallpox vaccines evaluation.

    PubMed

    Leparc-Goffart, Isabelle; Poirier, Bertrand; Garin, Daniel; Tissier, Marie-Hélène; Fuchs, Florence; Crance, Jean-Marc

    2005-01-01

    The possibility of mass population vaccination with smallpox vaccine implies the development of anti-vaccinia immunoglobulins for the treatment of severe side effects following vaccination. We have chosen to develop and validate the "gold standard method" (plaque reduction neutralization assay) to titrate neutralizing anti-vaccinia antibodies in two different French laboratories belonging to the Department of Defense (CRSSA) and to the French Health Products Safety Agency (Afssaps). The results of precision, linearity and accuracy of the method led to consider the method as validated. In parallel, we have prepared and lyophilized a pool of anti-vaccinia plasma samples issued from a unique donor and qualified this preparation versus the first British standard to use it as an in-house standard with a titer of 25 international units (IU). This work will allow to titrate, in IU, sera from vaccinated persons in order (i) to titrate purified anti-vaccinia immunoglobulin preparations for vaccine severe side effect treatments; (ii) to investigate the level of neutralizing antibodies in the general population; and (iii) to investigate clinical trials of new generation smallpox vaccines. In the future, this will allow comparability of studies on either smallpox vaccines or on the serological status of the population.

  3. Utilizing the virus-induced blocking of apoptosis in an easy baculovirus titration method.

    PubMed

    Niarchos, Athanasios; Lagoumintzis, George; Poulas, Konstantinos

    2015-01-01

    Baculovirus-mediated protein expression is a robust experimental technique for producing recombinant higher-eukaryotic proteins because it combines high yields with considerable post-translational modification capabilities. In this expression system, the determination of the titer of recombinant baculovirus stocks is important to achieve the correct multiplicity of infection for effective amplification of the virus and high expression of the target protein. To overcome the drawbacks of existing titration methods (e.g., plaque assay, real-time PCR), we present a simple and reliable assay that uses the ability of baculoviruses to block apoptosis in their host cells to accurately titrate virus samples. Briefly, after incubation with serial dilutions of baculovirus samples, Sf9 cells were UV irradiated and, after apoptosis induction, they were viewed via microscopy; the presence of cluster(s) of infected cells as islets indicated blocked apoptosis. Subsequently, baculovirus titers were calculated through the determination of the 50% endpoint dilution. The method is simple, inexpensive, and does not require unique laboratory equipment, consumables or expertise; moreover, it is versatile enough to be adapted for the titration of every virus species that can block apoptosis in any culturable host cells which undergo apoptosis under specific conditions.

  4. Amperometric, Bipotentiometric, and Coulometric Titration.

    ERIC Educational Resources Information Center

    Stock, John T.

    1984-01-01

    Reviews literature on amperometric, bipotentiometric, and coulometric titration methods examining: apparatus and methodology; acid-base reactions; precipitation and complexing reactions (considering methods involving silver, mercury, EDTA or analogous reagents, and other organic compounds); and oxidation-reduction reactions (considering methods…

  5. Estimating iron and aluminum content of acid mine discharge from a north-central Pennsylvania coal field by use of acidity titration curves

    USGS Publications Warehouse

    Ott, A.N.

    1986-01-01

    Determination of acidity provides a value that denotes the quantitative capacity of the sample water to neutralize a strong base to a particular pH. However, much additional information can be obtained from this determination if a titration curve is constructed from recorded data of titrant increments and their corresponding pH values. The curve can be used to identify buffer capabilities, the acidity with respect to any pH value within the curve limit, and, in the case of acid mine drainage from north-central Pennsylvania, the identification and estimation of the concentration of dissolved ferrous iron, ferric iron, and aluminum. Through use of titration curves, a relationship was observed for the acid mine drainage between: (1) the titratable acidity (as milligrams per liter calcium carbonate) to pH 4.0 and the concentration of dissolved ferric iron; and (2) the titratable acidity (as milligrams per liter calcium carbonate) from pH 4.0 to 5.0 and the concentration of dissolved aluminum. The presence of dissolved ferrous iron can be detected by the buffering effect exhibited in the area between pH 5.5 to 7.5. The concentration of ferrous iron is estimated by difference between the concentrations of ferric iron in an oxidized and unoxidized sample. Interferences in any of the titrations from manganese, magnesium, and aluminate, appear to be negligible within the pH range of interest.

  6. A new numerical method of finding potentiometric titration end-points by use of rational spline functions.

    PubMed

    Ren, K; Ren-Kurc, A

    1986-08-01

    A new numerical method of determining the position of the inflection point of a potentiometric titration curve is presented. It consists of describing the experimental data (emf, volume data-points) by means of a rational spline function. The co-ordinates of the titration end-point are determined by analysis of the first and second derivatives of the spline function formed. The method also allows analysis of distorted titration curves which cannot be interpreted by Gran's or other computational methods. PMID:18964159

  7. A rapid method for infectivity titration of Andes hantavirus using flow cytometry.

    PubMed

    Barriga, Gonzalo P; Martínez-Valdebenito, Constanza; Galeno, Héctor; Ferrés, Marcela; Lozach, Pierre-Yves; Tischler, Nicole D

    2013-11-01

    The focus assay is currently the most commonly used technique for hantavirus titer determination. This method requires an incubation time of between 5 and 11 days to allow the appearance of foci after several rounds of viral infection. The following work presents a rapid Andes virus (ANDV) titration assay, based on viral nucleocapsid protein (N) detection in infected cells by flow cytometry. To this end, an anti-N monoclonal antibody was used that was developed and characterized previously. ANDV N could be detected as early as 6 h post-infection, while viral release was not observed until 24-48 h post-infection. Given that ANDV detection was performed during its first round of infection, a time reduction for titer determination was possible and provided results in only two days. The viral titer was calculated from the percentage of N positive cells and agreed with focus assay titers. Furthermore, the assay was applied to quantify the inhibition of ANDV cell entry by patient sera and by preventing endosome acidification. This novel hantavirus titration assay is a highly quantitative and sensitive tool that facilitates infectivity titration of virus stocks, rapid screening for antiviral drugs, and may be further used to detect and quantify infectious virus in human samples.

  8. Acid-base equilibria in ethylene glycol--III: selection of titration conditions in ethylene glycol medium, protolysis constants of alkaloids in ethylene glycol and its mixtures.

    PubMed

    Zikolov, P; Zikolova, T; Budevsky, O

    1976-08-01

    Theoretical titration curves are used for the selection of appropriate conditions for the acid-base volumetric determination of weak bases in ethylene glycol medium. The theoretical curves for titration of some alkaloids are deduced graphically on the basis of the logarithmic concentration diagram. The acid-base constants used for the construction of the theoretical titration curves were determined by potentiometric titration in a cell without liquid junction, equipped with a glass and a silver-silver chloride electrode. It is shown that the alkaloids investigated can be determined accurately by visual or potentiometric titration. The same approach for the selection of titration conditions seems to be applicable to other non-aqueous amphiprotic solvents.

  9. Simple sensor for potentiometric titrations

    SciTech Connect

    Selig, W.

    1982-01-01

    A sensor for potentiometric titrations was prepared by coating a spectroscopic graphite rod with a solution of poly(vinyl chloride) and dioctylphthalate in tetrahydrofuran. The reference was an Ag/AgCl single-junction electrode. The sensor was used in the following potentiometric titrations: (1) precipitation titrations, (2) acid-base titrations, (3) compleximetric titrations, and (4) redox titrations. A survey of its use in such titrations is presented. Preparation of the coated-graphite sensor is simple and rapid. Moreover, it is quite inexpensive. A limitation is its applicability in aqueous media only, because organic solvents will dissolve the membrane. 5 figures, 4 tables.

  10. Titrations using an apparatus for recording the antilogarithm of pH or pM.

    PubMed

    Johansson, A

    1974-12-01

    A titration apparatus plotting either the concentration of sample ion or the concentration of titrant ion has been tested. An antilog apparatus, converting measured e.m.f. values into concentrations is connected to ordinary titration equipment. The instrument has been tested by means of acid-base titrations (titrations of mixtures of weak acids and of a weak and a strong acid), precipitation titrations (determination of the chloride concentration in tap-water, titration of mixtures of halides), titrations with ion-selective electrodes (determination of the fluoride content of toothpaste) and complexometric titrations (determination of copper with EDTA, using mercuric ion as indicator ion and amalgamated silver rod as indicator electrode, or using a copper-selective indicator electrode). The method considerably simplifies the evaluation of the results as compared to conventional potentiometric titrations.

  11. Surface characteristic changes of dental ceramics after cyclic immersion in acidic agents and titratable acidity.

    PubMed

    Junpoom, Peerapong; Kukiattrakoon, Boonlert; Hengtrakool, Chanothai

    2010-12-01

    The potential erosive effect of acidic food, sour fruits and drinks on all-ceramic restorations used in dentistry has not been clearly documented. Surface characteristic changes have been evaluated and compared for disc-shaped specimens (diameter 12.0 mm and thickness 2.0 mm) of fluorapatite-leucite and fluorapatite ceramics using various storage agents (deionized water, citrate buffer solution, pineapple juice, green mango juice, cola soft drink and 4% acetic acid). Immersion in pineapple juice, green mango juice, cola soft drink and 4% acetic acid for 16 hours produce significant increases in surface roughness for both types of ceramics investigated.

  12. Prediction of uranium and technetium sorption during titration of contaminated acidic groundwater

    SciTech Connect

    Zhang, Fan; Parker, Jack C.; Watson, David B; Jardine, Philip M; Gu, Baohua

    2010-01-01

    This study investigates uranium and technetium sorption onto aluminum and iron hydroxides during titration of acidic groundwater. The contaminated groundwater exhibits oxic conditions with high concentrations of NO{sub 3}{sup -}, SO{sub 4}{sup 2-}, U, Tc, and various metal cations. More than 90% of U and Tc was removed from the aqueous phase as Al and Fe precipitated above pH 5.5, but was partially resolublized at higher pH values. An equilibrium hydrolysis and precipitation reaction model adequately described variations in aqueous concentrations of metal cations. An anion exchange reaction model was incorporated to simulate sulfate, U and Tc sorption onto variably charged (pH-dependent) Al and Fe hydroxides. Modeling results indicate that competitive sorption/desorption on mixed mineral phases needs to be considered to adequately predict U and Tc mobility. The model could be useful for future studies of the speciation of U, Tc and co-existing ions during pre- and post-groundwater treatment practices.

  13. Influenza virus titration, antigenic characterization, and serological methods for antibody detection.

    PubMed

    Klimov, Alexander; Balish, Amanda; Veguilla, Vic; Sun, Hong; Schiffer, Jarad; Lu, Xiuhua; Katz, Jacqueline M; Hancock, Kathy

    2012-01-01

    This chapter describes some commonly used methods of influenza virus titration, antigenic characterization, and serological methods by antibody detection. These methods are essential not only for virus characterization but also for identifying new antigenic variants, vaccine strain selection, and sero-epidemiologic studies of influenza virus transmission and prevalence. Virus titration methods such as the hemagglutination assay, 50% egg or tissue culture infectious dose, and plaque assay are employed to determine the amount of virus particles in a sample. The hemagglutination inhibition assay is a reliable, relatively simple and inexpensive technique to antigenically characterize isolates of influenza viruses. Serological methods such as virus neutralization and hemagglutination inhibition are the fundamental tools used in sero-epidemiologic studies of influenza virus transmission and prevalence and in the evaluation of vaccine immunogenicity. While serological methods rarely yield an early diagnosis of acute influenza virus infection, well-timed, paired acute, and convalescent serum samples may establish the diagnosis of a recent influenza infection even when attempts to detect the virus are negative.

  14. Protonated polynucleotides structures - 22.CD study of the acid-base titration of poly(dG).poly(dC).

    PubMed

    Marck, C; Thiele, D; Schneider, C; Guschlbauer, W

    1978-06-01

    The acid-base titration (pH 8 --> pH 2.5 --> pH 8) of eleven mixing curve samples of the poly(dG) plus poly(dC) system has been performed in 0.15 M NaCl. Upon protonation, poly(dG).poly(dC) gives rise to an acid complex, in various amounts according to the origin of the sample. We have established that the hysteresis of the acid-base titration is due to the non-reversible formation of an acid complex, and the liberation of the homopolymers at the end of the acid titration and during the base titration: the homopolymer mixtures remain stable up to pH 7. A 1G:1C stoichiometry appears to be the most probable for the acid complex, a 1G:2C stoichiometry, as found in poly(C(+)).poly(I).poly(C) or poly(C(+)).poly(G).poly(C), cannot be rejected. In the course of this study, evidence has been found that the structural consequences of protonation could be similar for both double stranded poly(dG).poly(dC) and G-C rich DNA's: 1) protonation starts near pH 6, dissociation of the acid complex of poly(dG).poly(dC) and of protonated DNA take place at pH 3; 2) the CD spectrum computed for the acid polymer complex displays a positive peak at 255 nm as found in the acid spectra of DNA's; 3) double stranded poly(dG).poly(dC) embedded in triple-stranded poly(dG).poly(dG).poly(dC) should be in the A-form and appears to be prevented from the proton induced conformational change. The neutral triple stranded poly(dG).poly(dG).poly(dC) appears therefore responsible, although indirectly, for the complexity and variability of the acid titration of poly(dG).poly(dC) samples.

  15. Net proton-hydroxyl permeability of large unilamellar liposomes measured by an acid-base titration technique.

    PubMed

    Nichols, J W; Deamer, D W

    1980-04-01

    The net proton-hydroxyl permeability of large unilamellar liposomes has been measured by an acid-base pulse titration technique and has been determined to be several orders of magnitude greater than that measured for other monovalent ions. This permeability is relatively insensitive to variations in lipid composition. Proton permeability and hydroxyl permeability vary with pH 6 to 8, and this variation can occur in the absence of alterations in surface charge density resulting from titrations of acidic and basic groups on the lipids. In order to account for the exceptionally high proton-hydroxyl permeability with respect to other monovalent ions, we propose that protons or hydroxyls or both interact with clusters of hydrogen-bonded water molecules in the lipid bilayer, such that they are transferred across the bilayer by rearrangement of hydrogen bonds in a manner similar to their transport in water and ice.

  16. A potential high-throughput method for the determination of lipase activity by potentiometric flow injection titrations.

    PubMed

    Vahl, Katja; Kahlert, Heike; Böttcher, Dominique; Wardenga, Rainer; Komorsky-Lovrić, Sebojka; Bornscheuer, Uwe; Scholz, Fritz

    2008-03-01

    Potentiometric FIA titrations were performed to determine enzyme activities of lipase type B from Candida antarctica, CAL-B. Two substrates, triacetin and tributyrin were hydrolyzed in phosphate buffer solutions, and the concentration change of the base component of the buffer was titrated in a carrier solution containing hydrochloric acid and potassium chloride. The system was calibrated with butyric acid and acetic acid, respectively. FIA titration peaks were evaluated with respect to peak height and peak area. Butyric acid and acetic acid could be titrated in the buffer solution from 3x10(-3) mol L(-1) to 0.1 mol L(-1). The detection limit of enzyme activity was determined to be 0.07 U mL(-1) (15 min reaction time) and the minimum activity was calculated to be 0.035 units corresponding to 35 nmol min(-1). The specific activities of lipase B for the hydrolysis of tributyrin and triacetin were determined as 16+/-2 U mg(-1) and 2+/-0.2 U mg(-1) (per mg commercial lipase preparation), respectively.

  17. [Semiautomatic potentiometric titration of gastric juice].

    PubMed

    Mnuskina, M M; Meerov, G I; Nadezhina, L S; Semenov, V P; Chekanina, M I; Savchenko, V P; Grinzaĭd, E L

    1990-01-01

    Using commercial equipment manufactured in this country, the authors have assembled a unit for semiautomatic potentiometric titration and developed a method for such titration, permitting a rapid determination of free and bound HCl and the total acidity of the gastric juice. The equipment and method are recommended for wide use at laboratories of therapeutic institutions, of biologic factories manufacturing medicinal commercial gastric juice, and of research institutes.

  18. Determination of total fluoride in HF/HNO3/H2SiF6 etch solutions by new potentiometric titration methods.

    PubMed

    Weinreich, Wenke; Acker, Jörg; Gräber, Iris

    2007-03-30

    In the photovoltaic industry the etching of silicon in HF/HNO(3) solutions is a decisive process for cleaning wafer surfaces or to produce certain surface morphologies like polishing or texturization. With regard to cost efficiency, a maximal utilisation of etch baths in combination with highest quality and accuracy is strived. To provide an etch bath control realised by a replenishment with concentrated acids the main constituents of these HF/HNO(3) etch solutions including the reaction product H(2)SiF(6) have to be analysed. Two new methods for the determination of the total fluoride content in an acidic etch solution based on the precipitation titration with La(NO(3))(3) are presented within this paper. The first method bases on the proper choice of the reaction conditions, since free fluoride ions have to be liberated from HF and H(2)SiF(6) at the same time to be detected by a fluoride ion-selective electrode (F-ISE). Therefore, the sample is adjusted to a pH of 8 for total cleavage of the SiF(6)(2-) anion and titrated in absence of buffers. In a second method, the titration with La(NO(3))(3) is followed by a change of the pH-value using a HF resistant glass-electrode. Both methods provide consistent values, whereas the analysis is fast and accurate, and thus, applicable for industrial process control. PMID:19071540

  19. Aggregation property of glycyrrhizic acid and its interaction with cyclodextrins analyzed by dynamic light scattering, isothermal titration calorimetry, and NMR.

    PubMed

    Izutani, Yusuke; Kanaori, Kenji; Oda, Masayuki

    2014-06-17

    The structural properties of glycyrrhizic acid, a sweet-tasting constituent of Glycyrrhiza glabra, and its interaction with cyclodextrins were analyzed using dynamic light scattering, isothermal titration calorimetry, and NMR. The dynamic light scattering and NMR studies showed that glycyrrhizic acid forms a water-soluble aggregate that disperses upon the addition of γ-cyclodextrin. The high sweetness of glycyrrhizic acid can be closely correlated with this aggregation, because the multimers of glycyrrhizic acid can simultaneously bind to the sweet taste receptors on the human tongue. The isothermal titration calorimetry experiments demonstrated that γ-cyclodextrin binds to glycyrrhizic acid more strongly than β-cyclodextrin, however, both reactions are accompanied by a favorable change in binding entropy. Considering the large negative change in heat capacity that is observed during the binding of γ-cyclodextrin, the main driving force for the binding is hydrophobic interactions with dehydration, which is typical for inclusion complex. NMR experiments showed that γ-cyclodextrin interacts with the central part of the aglycone moiety, not the glucuronic acid moieties, resulting in high binding affinity. It should also be noted that the two distinct complexes of glycyrrhizic acid with γ-cyclodextrin would exist in aqueous solution.

  20. Development of a novel baculovirus titration method using the Enzyme-linked immunosorbent spot (ELISPOT) assay.

    PubMed

    Wang, Wei; Cheng, Tong; Ma, Ke; Xia, Dezhen; Wang, Yongmei; Liu, Jian; Du, Hailian; Shih, James Wai Kuo; Zhang, Jun; Zhao, Qinjian; Xia, Ningshao

    2013-03-01

    The baculovirus expression vector system (BEVS) is one of the most powerful methods for production of recombinant proteins for research or commercial purposes. Titration of viable virus in insect cell culture is often required when BEVS is used for basic research or bioprocessing. An enzyme-linked immunosorbent spot (ELISPOT) assay using monoclonal antibodies against the major capsid protein VP39 of both Autographa californica nuclear polyhedrosis virus (AcMNPV) and Bombyx mori nuclear polyhedrosis virus (BmNPV) was developed for baculovirus quantitation at 48h post-infection. The titer was determined by visualizing infected insect cells as blue spots and automated spot counting was achieved with ELISPOT hardware and software. Log-scale comparison of the results between the ELISPOT assay and a conventional end point dilution assay using a fluorescent marker showed a good correlation for both AcMNPV (R(2)=0.9980, p<0.05) and BmNPV (R(2)=0.9834, p<0.05). In conclusion, a novel, rapid and semi-automated procedure for titrating baculovirus was developed based on the specific immunostaining of infected cells followed by automated spot counting.

  1. A comparison of titration methods for live avian encephalomyelitis virus vaccines.

    PubMed

    Nicholas, R A; Hopkins, I G; Southern, S J; Thornton, D H

    1986-01-01

    Factors associated with the indirect fluorescent antibody test used for the titration of avian encephalomyelitis virus (AEV) in chick embryo brain cell cultures were examined for their influence on virus replication. It was found that virus should be inoculated onto semi-confluent cell cultures and adsorbed for two hours at room temperature. The cells should then be examined for fluorescence after five days' incubation. Using these conditions, the cell culture assay was compared with the embryo and chick assays for its ability to estimate the virus content of live commercial AEV vaccines. In most cases titres obtained by the chick assay were slightly, but not significantly, higher than those obtained in the cell culture assay, although the reliability of the chick assay was, at times, questionable. In all cases titres obtained in the embryo assay were low. It is recommended that the cell culture assay be adopted as the method of choice for titrating AEV vaccines because it is rapid, reproducible, specific and greatly reduces the requirement for experimental animals.

  2. Drug-binding energetics of human α-1-acid glycoprotein assessed by isothermal titration calorimetry and molecular docking simulations

    PubMed Central

    Huang, Johnny X.; Cooper, Matthew A.; Baker, Mark A.; Azad, Mohammad A.K.; Nation, Roger L.; Li, Jian; Velkov, Tony

    2012-01-01

    This study utilizes sensitive, modern isothermal titration calorimetric (ITC) methods to characterize the microscopic thermodynamic parameters that drive the binding of basic drugs to α-1-acid glycoprotein (AGP) and thereby rationalize the thermodynamic data in relation to docking models and crystallographic structures of the drug-AGP complexes. The binding of basic compounds from the tricyclic antidepressant series, together with miaserine, chlorpromazine, disopyramide and cimetidine all displayed an exothermically driven binding interaction with AGP. The impact of protonation/deprotonation events, ionic strength, temperature and the individual selectivity of the A and F1*S AGP variants on drug-binding thermodynamics were characterized. A correlation plot of the thermodynamic parameters for all of the test compounds revealed enthalpy-entropy compensation is in effect. The exothermic binding energetics of the test compounds were driven by a combination of favorable (negative) enthalpic (ΔH°) and favorable (positive) entropic (ΔS°) contributions to the Gibbs free energy (ΔG°). Collectively, the data imply that the free energies that drive drug binding to AGP and its relationship to drug-serum residency evolve from the complex interplay of enthalpic and entropic forces from interactions with explicit combinations of hydrophobic and polar side-chain sub-domains within the multi-lobed AGP ligand binding cavity. PMID:23192962

  3. Dye uptake assay: an efficient and sensitive method for human interferon titration.

    PubMed

    Pidot, A L

    1971-10-01

    Currently, human interferon (IF) assays are generally performed by plaque reduction or visual cytopathic effect methods. Both are time-consuming, subjective in interpretation, and, in the case of the latter, relatively insensitive. An adaption of the dye uptake method for human IF titration which uses foreskin-derived fibroblasts and vesicular stomatitis virus is described. This assay is reproducible and sensitive (1 unit = 3 international units). By direct comparison, however, it is somewhat less sensitive than the plaque reduction assay (1 unit = 1.1 international units). This assay is especially recommended for use in needed clinical investigations of human IF because of its technical simplicity, allowing efficient handling of large numbers of specimens.

  4. A method of alphavirus replicon particle titration based on expression of functional replicase/transcriptase.

    PubMed

    Gangolli, Seema S; Vasilakis, Nikolaos; Kovacs, Gerald R; Zamb, Timothy J; Kowalski, Jacek

    2003-05-01

    Alphavirus replicon particles are being exploited for a variety of purposes both in vitro as gene expression vectors, and in vivo as vaccines or gene therapy vectors. There is a need for a simple and universal method of titration of replicon particles that is independent of expression of the foreign protein. We devised a method that uses modified vaccinia virus Ankara (MVA) as an indicator virus, to deliver a Venezuelan equine encephalitis virus (VEE) defective helper RNA encoding green fluorescent protein (GFP). Co-infection of cells with the MVA-based indicator and Venezuelan equine encephalitis virus replicon particles (VRP) results in expression of the GFP gene. VRP titer is readily determined by counting fluorescent cells.

  5. [Rigorous algorithms for calculating the exact concentrations and activity levels of all the different species during acid-base titrations in water].

    PubMed

    Burgot, G; Burgot, J L

    2000-10-01

    The principles of two algorithms allowing the calculations of the concentration and activity levels of the different species during acid-base titrations in water are described. They simulate titrations at constant and variable ionic strengths respectively. They are designed so acid and base strengths, their concentrations and the titrant volume added can be chosen freely. The calculations are based on rigorous equations with a general scope. They are sufficiently compact to be processed on pocket calculators. The algorithms can easily simulate pH-metric, spectrophotometric, conductometric and calorimetric titrations, and hence allow determining concentrations and some physico-chemical constants related to the occurring chemical systems.

  6. A one-step colorimetric acid-base titration sensor using a complementary color changing coordination system.

    PubMed

    Cho, Hui Hun; Kim, Si Hyun; Heo, Jun Hyuk; Moon, Young Eel; Choi, Young Hun; Lim, Dong Cheol; Han, Kwon-Hoon; Lee, Jung Heon

    2016-06-21

    We report the development of a colorimetric sensor that allows for the quantitative measurement of the acid content via acid-base titration in a single-step. In order to create the sensor, we used a cobalt coordination system (Co-complex sensor) that changes from greenish blue colored Co(H2O)4(OH)2 to pink colored Co(H2O)6(2+) after neutralization. Greenish blue and pink are two complementary colors with a strong contrast. As a certain amount of acid is introduced to the Co-complex sensor, a portion of greenish blue colored Co(H2O)4(OH)2 changes to pink colored Co(H2O)6(2+), producing a different color. As the ratio of greenish blue and pink in the Co-complex sensor is determined by the amount of neutralization reaction occurring between Co(H2O)4(OH)2 and an acid, the sensor produced a spectrum of green, yellow green, brown, orange, and pink colors depending on the acid content. In contrast, the color change appeared only beyond the end point for normal acid-base titration. When we mixed this Co-complex sensor with different concentrations of citric acid, tartaric acid, and malic acid, three representative organic acids in fruits, we observed distinct color changes for each sample. This color change could also be observed in real fruit juice. When we treated the Co-complex sensor with real tangerine juice, it generated diverse colors depending on the concentration of citric acid in each sample. These results provide a new angle on simple but quantitative measurements of analytes for on-site usage in various applications, such as in food, farms, and the drug industry. PMID:27143645

  7. A one-step colorimetric acid-base titration sensor using a complementary color changing coordination system.

    PubMed

    Cho, Hui Hun; Kim, Si Hyun; Heo, Jun Hyuk; Moon, Young Eel; Choi, Young Hun; Lim, Dong Cheol; Han, Kwon-Hoon; Lee, Jung Heon

    2016-06-21

    We report the development of a colorimetric sensor that allows for the quantitative measurement of the acid content via acid-base titration in a single-step. In order to create the sensor, we used a cobalt coordination system (Co-complex sensor) that changes from greenish blue colored Co(H2O)4(OH)2 to pink colored Co(H2O)6(2+) after neutralization. Greenish blue and pink are two complementary colors with a strong contrast. As a certain amount of acid is introduced to the Co-complex sensor, a portion of greenish blue colored Co(H2O)4(OH)2 changes to pink colored Co(H2O)6(2+), producing a different color. As the ratio of greenish blue and pink in the Co-complex sensor is determined by the amount of neutralization reaction occurring between Co(H2O)4(OH)2 and an acid, the sensor produced a spectrum of green, yellow green, brown, orange, and pink colors depending on the acid content. In contrast, the color change appeared only beyond the end point for normal acid-base titration. When we mixed this Co-complex sensor with different concentrations of citric acid, tartaric acid, and malic acid, three representative organic acids in fruits, we observed distinct color changes for each sample. This color change could also be observed in real fruit juice. When we treated the Co-complex sensor with real tangerine juice, it generated diverse colors depending on the concentration of citric acid in each sample. These results provide a new angle on simple but quantitative measurements of analytes for on-site usage in various applications, such as in food, farms, and the drug industry.

  8. A Unified Titration Formula

    NASA Astrophysics Data System (ADS)

    Glaister, P.

    1999-01-01

    In a number of standard titrations, a volume of a monoprotic base MOH at a specific concentration is added to a volume of a monoprotic acid HA at a specific concentrations. Four different types of titration are possible, depending on whether the acid and base are strong or weak. In a recent article, some of these cases are discussed in detail. However, a single unifying formula covering all four cases has been determined.

  9. Titration of a Solid Acid Monitored by X-Ray Diffraction

    ERIC Educational Resources Information Center

    Dungey, Keenan E.; Epstein, Paul

    2007-01-01

    An experiment is described to introduce students to an important class of solid-state reactions while reinforcing concepts of titration by using a pH meter and a powder X-ray diffractometer. The experiment was successful in teaching students the abstract concepts of solid-state structure and diffraction by applying the diffraction concepts learned…

  10. Comparative methods of titration of Silverwater virus, an arthropod borne virus.

    PubMed

    Belloncik, S; Boisvert, J

    1978-01-01

    Silverwater virus (SIL) was titrated in vivo and in vitro. Using a microculture technique, TCID50 was found to be much more sensitive than both the plaque forming assay and the LD50. The latter was the least sensitive mode of titration of Silverwater virus. The addition of DEAE dextran and trypsin to the viral inoculum increased the number and diameter of the plaques.

  11. Application of flow injection gradient titration based on the standard addition and dilution procedure to the determination of total acidity in vinegars and soft drinks.

    PubMed

    Wójtowicz, Marzena; Kozak, Joanna; Górnacka, Dorota; Koscielniak, Pawel

    2008-12-01

    The proposed approach relies on successive dilution of a solution containing a sample with standard addition and on titration of the solutions obtained until receiving a signal lower than the signal measured for the sample alone. Equations are derived for subsequent dilution factors for the automatic flow injection system applied. The overall calibration strategy including the possibility of prediction of the number of required standard solutions and of positioning the sample signal within the calibration range is presented. The method has been tested on the spectrophotometric determination of hydrochloric acid at two concentration levels (0.193 and 1.21 mol L(-1)) with sodium hydroxide as titrant. The results of accuracy better than |3.9|% (RE) and repeatability better than 1.3% (RSD) were obtained. The method has been applied to the determination of total acidity in vinegars and soft drinks. The average time necessary to analyze one sample is about 10 min. PMID:19075470

  12. Reference test methods for total water in lint cotton by Karl Fischer Titration and low temperature distillation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In a study of comparability of total water contents (%) of conditioned cottons by Karl Fischer Titration (KFT) and Low Temperature Distillation (LTD) reference methods, we demonstrated a match of averaged results based on a large number of replications and weighing the test specimens at the same tim...

  13. Effects of dairy factory, milk casein content and titratable acidity on coagulation properties in Trentingrana dairy industry.

    PubMed

    Penasa, Mauro; Toffanin, Valentina; Cologna, Nicola; Cassandro, Martino; De Marchi, Massimo

    2016-05-01

    The objective of the present study was to investigate the effect of environmental factors, milk casein content and titratable acidity on milk coagulation properties (MCP) of samples routinely collected in the Trento province (northeast Italy) under field conditions. Rennet coagulation time (RCT, min), curd-firming time (k20, min) and curd firmness (a30, mm) were determined by Formagraph on 14 971 samples from 635 herds associated to 17 dairy factories. Besides MCP, fat, protein, and casein percentages, titratable acidity (TA), and somatic cell and bacterial counts were available. A standardised index of milk aptitude to coagulate (IAC) was derived using information of RCT and a30. An analysis of variance was conducted on MCP and IAC using a fixed effects linear model. Approximately 3% of milk samples did not form a curd within the testing time (30 min) and k20 was missing for 26% of milks. The percentage of samples without information on k20 largely differed among dairy factories (1·7-20·9%). Significant differences were estimated between the best and the worst dairy factory for RCT (-2 min), k20 (-1·2 min), a30 (+3·4 mm) and IAC (+2·6 points). Milk casein content and TA were important factors in explaining the variation of MCP and IAC, supporting the central role of these two traits on technological properties. The Trento province is heterogeneous in terms of dairy systems and this could explain the differences among dairy factories.

  14. Effects of dairy factory, milk casein content and titratable acidity on coagulation properties in Trentingrana dairy industry.

    PubMed

    Penasa, Mauro; Toffanin, Valentina; Cologna, Nicola; Cassandro, Martino; De Marchi, Massimo

    2016-05-01

    The objective of the present study was to investigate the effect of environmental factors, milk casein content and titratable acidity on milk coagulation properties (MCP) of samples routinely collected in the Trento province (northeast Italy) under field conditions. Rennet coagulation time (RCT, min), curd-firming time (k20, min) and curd firmness (a30, mm) were determined by Formagraph on 14 971 samples from 635 herds associated to 17 dairy factories. Besides MCP, fat, protein, and casein percentages, titratable acidity (TA), and somatic cell and bacterial counts were available. A standardised index of milk aptitude to coagulate (IAC) was derived using information of RCT and a30. An analysis of variance was conducted on MCP and IAC using a fixed effects linear model. Approximately 3% of milk samples did not form a curd within the testing time (30 min) and k20 was missing for 26% of milks. The percentage of samples without information on k20 largely differed among dairy factories (1·7-20·9%). Significant differences were estimated between the best and the worst dairy factory for RCT (-2 min), k20 (-1·2 min), a30 (+3·4 mm) and IAC (+2·6 points). Milk casein content and TA were important factors in explaining the variation of MCP and IAC, supporting the central role of these two traits on technological properties. The Trento province is heterogeneous in terms of dairy systems and this could explain the differences among dairy factories. PMID:27210496

  15. Analysis of potentiometric titrations of heterogeneous natural polyelectrolytes in terms of counterion condensation theory: application to humic acid.

    PubMed

    Porasso, R D; Benegas, J C; van den Hoop, M; Paoletti, S

    2000-07-31

    A model, developed within the framework of the counterion condensation theory of linear polyelectrolytes, is presented in this paper to describe the acid-base properties of linear polyelectrolytes, consisting of several types of functional ionizable groups. This formalism has been successfully applied to Fluka humic acid under salt-free conditions, as well as in the presence of supporting simple 1:1 salt (KNO3) at three different concentrations. As part of this approach, the charge density of the humic acid is obtained from the activity coefficient measurements of potassium counterions at different humic acid concentrations at a constant degree of dissociation of the polyelectrolyte. The humic acid average charge density was found to be 0.80 +/- 0.05. Using the present model, we are able to satisfactorily describe the experimental data obtained from acid-base potentiometric titrations. Four main functional groups making up the polymer are determined through their fractional abundances (Xi) and intrinsic pK (pK0i) values. The fractional abundances remained constant and independent of the ionic strength, indicating that the humic acid constitution does not depend on the concentration of excess salts. The pK0i values show a small change with ionic strength, which can be explained by the polyelectrolytic behavior of the solution.

  16. The interaction of phenolic acids with Fe(III) in the presence of citrate as studied by isothermal titration calorimetry.

    PubMed

    Yang, Senpei; Bai, Guangling; Chen, Lingli; Shen, Qun; Diao, Xianmin; Zhao, Guanghua

    2014-08-15

    Under physiological conditions, exogenous chelators such as polyphenols might interact with non-protein bound ferric complexes, such as Fe(III)-citrate. Additionally, Fe(III) and citrate are widely distributed in various fruits and vegetables which are also rich in phenolic acids. In this study, we focus on the interaction between phenolic acids (gallic acid, methyl gallate and protocatechuic acid) and Fe(III) in the presence of excessive citrate by isothermal titration calorimetry (ITC) for thermodynamic studies, and stopped-flow absorption spectrometry for fast kinetic studies. Results reveal that all of these three phenolic acids can bind to the Fe(III) with the same stoichiometry (3:1). Moreover, the binding constants of these three compounds with Fe(III) are greatly dependent on ligand structure, and are much higher than that of Fe(III)-citrate. Based on their stoichiometry and superhigh binding constants, it is most likely that these three phenolic acids can displace the citrate to bind with one iron(III) ion to form a stable octahedral geometric structure, albeit at different rates. These findings shed light on the interaction between phenolic acids and Fe(III) in the presence of citrate under either physiological conditions or in a food system.

  17. Analysis of the sorption properties of different soils using water vapour adsorption and potentiometric titration methods

    NASA Astrophysics Data System (ADS)

    Skic, Kamil; Boguta, Patrycja; Sokołowska, Zofia

    2016-07-01

    Parameters of specific surface area as well as surface charge were used to determine and compare sorption properties of soils with different physicochemical characteristics. The gravimetric method was used to obtain water vapour isotherms and then specific surface areas, whereas surface charge was estimated from potentiometric titration curves. The specific surface area varied from 12.55 to 132.69 m2 g-1 for Haplic Cambisol and Mollic Gleysol soil, respectively, and generally decreased with pH (R=0.835; α = 0.05) and when bulk density (R=-0.736; α = 0.05) as well as ash content (R=-0.751; α = 0.05) increased. In the case of surface charge, the values ranged from 63.00 to 844.67 μmol g-1 Haplic Fluvisol and Mollic Gleysol, respecively. Organic matter gave significant contributions to the specific surface area and cation exchange capacity due to the large surface area and numerous surface functional groups, containing adsorption sites for water vapour molecules and for ions. The values of cation exchange capacity and specific surface area correlated linearly at the level of R=0.985; α = 0.05.

  18. Thermodynamics of ion binding to phosphatidic acid bilayers. Titration calorimetry of the heat of dissociation of DMPA.

    PubMed

    Blume, A; Tuchtenhagen, J

    1992-05-19

    The heat of dissociation of the second proton of 1,2-dimyristoylphosphatidic acid (DMPA) was studied as a function of temperature using titration calorimetry. The dissociation of the second proton of DMPA was induced by addition of NaOH. From the calorimetric titration experiment, the intrinsic pK0 for the dissociation reaction could be determined by applying the Gouy-Chapman theory. pK0 decreases with temperature from ca. 6.2 at 11 degrees C to 5.4 at 54 degrees C. From the total heat of reaction, the dissociation enthalpy, delta Hdiss, was determined by subtracting the heat of neutralization of water and the heat of dilution of NaOH. In the temperature range between 2 and 23 degrees C, delta Hdiss is endothermic with an average value of ca. 2.5 kcal.mol-1 and shows no clear-cut temperature dependence. In the temperature range between 23 and 52 degrees C, delta Hdiss calculated after subtraction of the heat of neutralization and dilution is not the true dissociation enthalpy but includes contributions from the phase transition enthalpy, delta Htrans, as the pH jump induces a transition from the gel to the liquid-crystalline phase. The delta Cp for the reaction enthalpy observed in this temperature range is positive. Above 53 degrees C, the pH jump induces again only the dissociation of the second proton, and the bilayers stay in the liquid-crystalline phase. In this temperature range, delta Hdiss seems to decrease with temperature. The thermodynamic data from titration calorimetry and differential scanning calorimetry as a function of pH can be combined to construct a complete enthalpy-temperature diagram of DMPA in its two ionization states.

  19. End-point dilution and plaque assay methods for titration of cricket paralysis virus in cultured Drosophila cells.

    PubMed

    Scotti, P D

    1977-05-01

    Cricket paralysis virus, an insect picorna-like virus, caused a distinct c.p.e. in cultured Drosophila melanogaster cells, allowing the development of titration methods based on end-point dilution or plaque assay methods. The end-point dilution (TCD50) method is more sensitive and economical than plaque assays and is easily scored. The data indicate a minimum infectivity/particle ratio of about 1/2000.

  20. Reverse plaque formation method for titration of non-cytopathogenic bovine viral diarrhea-mucosal disease virus.

    PubMed

    Itoh, O; Sasaki, H; Hanaki, T

    1983-01-01

    The reverse plaque formation (RPF) method with a semi-micro plate was applied to the titration of a non-cytopathogenic (non-CP) strain of bovine viral diarrhea-mucosal disease (BVD-MD) virus. All the five non-CP strains used in this experiment formed reverse plaques (RP) on bovine testicle cell culture under methyl cellulose overlay. The RPF was inhibited by the pretreatment of a non-CP virus strain with immune rabbit serum to a reference strain. The specificity of the RPF method was demonstrated by the linear test and Poisson distribution test. Comparative titration of commercial BVD-MD vaccines was carried out by the semi-micro RPF method and the tube method based on the exaltation on Newcastle disease virus. The virus titer obtained by the former was slightly higher than that obtained by the latter. The former was proved to be a method of high sensitivity for determining non-CP virus.

  1. Filtrates & Residues: Olfactory Titration.

    ERIC Educational Resources Information Center

    Wood, John T.; Eddy, Roberta M.

    1996-01-01

    Presents an experiment that uses a unique acid-base indicator--the odor of raw onion--to indicate the end point of the titration of sodium hydroxide with hydrochloric acid. Allows the student to detect the completion of the neutralization reaction by olfaction rather than sight. (JRH)

  2. Acid titrations of poly(dG-dC).poly(dG-dC) in aqueous solution and in a w/o microemulsion.

    PubMed

    Airoldi, Marta; Boicelli, C Andrea; Gennaro, Guiseppe; Giomini, Marcello; Giuliani, Anna Maria; Giustini, Mauro

    2006-02-01

    The model polynucleotide poly(dG-dC).poly(dG-dC) (polyGC) was titrated with a strong acid (HCl) in aqueous unbuffered solutions and in the quaternary w/o microemulsion CTAB/n-pentanol/n-hexane/water. The titrations, performed at several concentrations of NaCl in the range 0.005 to 0.600 M, were followed by recording the modifications of the electronic absorption and of the CD spectra (210< or = lambda < or =350 nm) upon addition of the acid. In solution, the polynucleotide undergoes two acid-induced transitions, neither of which corresponds to denaturation of the duplex to single coil. The first transition leads to the Hoogsteen type synG.C+ duplex, while the second leads to the C+.C duplex. The initial B-form of polyGC was recovered by back-titration with NaOH. The apparent pKa values were obtained for both steps of the titration, at all salt concentrations. A reasonably linear dependence of pKa1 and pKa2 from p[NaCl] was obtained, with both pKa values decreasing with increasing ionic strength. In microemulsion, at salt concentrations < or = 0.300 M, an acid-induced transition was observed, matching the first conformational transition recorded also in solution. However, further addition of acid led to denaturation of the protonated duplex. Renaturation of polyGC was obtained by back-titration with NaOH. At salt concentrations > 0.300 M, polyGC is present as a mixture of B-form and psi- aggregates, that slowly separate from the microemulsion. The acid titration induces at first a conformational transition similar to the one observed at low salt or in solution, then denaturation occurs, which is however preceded by the appearance of a transient conformation, that has been tentatively classified as a left-handed Z double helix.

  3. Kinetics and thermodynamics of interaction between sulfonamide antibiotics and humic acids: Surface plasmon resonance and isothermal titration microcalorimetry analysis.

    PubMed

    Xu, Juan; Yu, Han-Qing; Sheng, Guo-Ping

    2016-01-25

    The presence of sulfonamide antibiotics in the environments has been recognized as a crucial issue. Their migration and transformation in the environment is determined by natural organic matters that widely exist in natural water and soil. In this study, the kinetics and thermodynamics of interactions between humic acids (HA) and sulfamethazine (SMZ) were investigated by employing surface plasmon resonance (SPR) combined with isothermal titration microcalorimetry (ITC) technologies. Results show that SMZ could be effectively bound with HA. The binding strength could be enhanced by increasing ionic strength and decreasing temperature. High pH was not favorable for the interaction. Hydrogen bond and electrostatic interaction may play important roles in driving the binding process, with auxiliary contribution from hydrophobic interaction. The results implied that HA existed in the environment may have a significant influence on the migration and transformation of organic pollutants through the binding process.

  4. Surface properties of bacillus subtilis determined by acid/base titrations, and the implications for metal adsorption in fluid-rock systems

    SciTech Connect

    Fein, J.B.; Davis, T.A.

    1996-10-01

    Bacteria are ubiquitous in low temperature aqueous systems, but quantifying their effects on aqueous mass transport remains a problem. Numerous studies have qualitatively examined the metal binding capacity of bacterial cell walls. However, quantitative thermodynamic modeling of metal-bacteria-mineral systems requires a detailed knowledge of the surface properties of the bacterial functional groups. In this study, we have conducted acid/base titrations of suspensions of B. subtilis, a common subsurface species whose surface properties are largely controlled by carboxyl groups. Titrations were conducted between pH 2 and 11 at several ionic strengths. The data are analyzed using a constant capacitance model to account for the surface electric field effects on the acidity constant. The pK{sub a} value that best fits the titration data is 3.9 {plus_minus} 0.3. This result represents the first step toward quantifying bacteria-metal and mineral-bacteria-metal interactions using equilibrium thermodynamics.

  5. Titration of Newcastle disease virus and its neutralizing antibodies in microplates by a modified hemadsorption and hemadsorption inhibition method.

    PubMed

    Katz, D; Ben-Moshe, H; Alon, S

    1976-03-01

    Using the microtiter system, titration of Newcastle disease virus infectivity and neutralizing antibodies was carried out in chicken embryo fibroblasts grown in "U" or flat-bottomed plates. Infectivity was detected by a combined hemadsorption-hemagglutination method. Inhibition of that reaction indicated the presence of neutralizing antibodies. A 24-h microneutralization test was developed and compared to the plaque neutralization and microhemagglutination inhibition test. Reproducibility of the microneutralization test was statistically analyzed.

  6. Quantitative titration of nucleic acids by enzymatic amplification reactions run to saturation.

    PubMed Central

    Pannetier, C; Delassus, S; Darche, S; Saucier, C; Kourilsky, P

    1993-01-01

    In vitro enzymatic amplification of nucleic acids by PCR or other techniques is a very sensitive method to detect rare DNA segments. We present here a protocol that allows the rapid, sensitive and precise quantification of DNA molecules using PCR amplification run to saturation. The DNA (or cDNA) to be assayed is co-amplified with known amounts of an internal standard DNA. We show that the latter must be almost identical to the assayed DNA, otherwise quantification at the plateau is unreliable. The read-out of the amplification involves one or two additional oligonucleotides. Using fluorescent oligonucleotides as primers in run-off reactions together with an automated DNA sequencer, we could measure the level of expression of several genes, like the murine MHC class I H-2Kd or a specific T cell receptor beta chain transcript in the course of an immunization. mRNA levels were normalized by measuring in a similar manner the number of transcripts encoding the housekeeping gene HPRT. Finally, our procedure might allow the rapid analysis of a large number of samples at the same time, as illustrated by the simultaneous analysis of the mRNAs encoding the CD4 and CD8 murine T cell markers. PMID:8441670

  7. Quantification of Norwalk virus inocula: Comparison of endpoint titration and real-time reverse transcription-PCR methods.

    PubMed

    Liu, Pengbo; Hsiao, Hui-Mien; Jaykus, Lee-Ann; Moe, Christine

    2010-09-01

    Human noroviruses (NoV) are the leading cause of epidemic acute gastroenteritis. In order to fully characterize features such as persistence and infectious dose, precise quantification of virus concentration is necessary. The purpose of this study was to compare two methods [endpoint titration RT-PCR and quantitative RT-PCR (RT-qPCR)] with respect to quantification of Norwalk virus (NV) in inocula made from purified stock suspensions of human fecal specimens. A full-length NV RNA transcript was developed to facilitate quantification using RT-qPCR and provided log linear detection in the range of 49-4.9 x 10(4) genome equivalent copies (GEC) per reaction. Endpoint titration RT-PCR was used to estimate PCR detection units, and RT-qPCR was used to estimate genome copies in two NV inocula (8fIIa and 8fIIb) used in previous human challenge studies. Overall, RT-qPCR was 1.1-1.6 log(10) more sensitive (lower detection limit) than endpoint titration RT-PCR when the same RNA release method, PCR primers and thermocycle program were used. These findings have important implications for many experimental interpretations, not the least of which is estimating the median infectious dose in human challenge studies.

  8. Simple in vitro methods for titrating feline immunodeficiency virus (FIV) and FIV neutralizing antibodies.

    PubMed

    Tozzini, F; Matteucci, D; Bandecchi, P; Baldinotti, F; Poli, A; Pistello, M; Siebelink, K H; Ceccherini-Nelli, L; Bendinelli, M

    1992-06-01

    The feline immunodeficiency virus (FIV) readily produced syncytia in Crandell feline kidney (CrFK) cells adapted to a medium containing 0.5% fetal calf serum, a variety of growth factors and other supplements. This finding has been exploited to develop simple and sensitive virus titration and neutralization assays. High titre neutralizing antibodies were detected in cats infected naturally and experimentally with FIV, but not in uninfected animals.

  9. Digital image-based titrations.

    PubMed

    Gaiao, Edvaldo da Nobrega; Martins, Valdomiro Lacerda; Lyra, Wellington da Silva; de Almeida, Luciano Farias; da Silva, Edvan Cirino; Araújo, Mário César Ugulino

    2006-06-16

    The exploitation of digital images obtained from a CCD camera (WebCam) as a novel instrumental detection technique for titration is proposed for the first time. Named of digital image-based (DIB) titration, it also requires, as a traditional titration (for example, spectrophotometric, potentiometric, conductimetric), a discontinuity in titration curves where there is an end point, which is associated to the chemical equivalence condition. The monitored signal in the DIB titration is a RGB-based value that is calculated, for each digital image, by using a proposed procedure based on the red, green, and blue colour system. The DIB titration was applied to determine HCl and H3PO4 in aqueous solutions and total alkalinity in mineral and tap waters. Its results were compared to the spectrophotometric (SPEC) titration and, by applying the paired t-test, no statistic difference between the results of both methods was verified at the 95% confidence level. Identical standard deviations were obtained by both titrations in the determinations of HCl and H3PO4, with a slightly better precision for DIB titration in the determinations of total alkalinity. The DIB titration shows to be an efficient and promising tool for quantitative chemical analysis and, as it employs an inexpensive device (WebCam) as analytical detector, it offers an economically viable alternative to titrations that need instrumental detection.

  10. Determination of soluble solids content and titratable acidity of intact fruit and juice of Satsuma mandarin using a hand-held NIR instrument in transmittance mode

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Determination of soluble solids content (SSC) and titratable acidity (TA) of intact fruit and juice of Satsuma mandarin was investigated using a hand-held NIR instrument “NIR-Gun” in transmittance mode. The resulting calibration equation measured SSC of intact fruit and juice with a standard error o...

  11. Analysis of aqueous solutions by near-infrared spectrometry (NIRS) II. Titrations of weak and very weak acids with strong bases

    NASA Astrophysics Data System (ADS)

    Molt, K.; Niemöller, A.; Cho, Y. J.

    1997-06-01

    The titrations of acetic and phosphoric acid with NaOH were observed by NIR-spectrometry. The measured spectra can be evaluated quantitatively for calibration of the H +-and OH --concentration. The NIRS errors with respect to the prediction of the pH and the equivalence point are discussed.

  12. Volumetric Titrations Using Electrolytically Generated Reagents for the Determination of Ascorbic Acid and Iron in Dietary Supplement Tablets: An Undergraduate Laboratory Experiment

    ERIC Educational Resources Information Center

    Scanlon, Christopher; Gebeyehu, Zewdu; Griffin, Kameron; Dabke, Rajeev B.

    2014-01-01

    An undergraduate laboratory experiment for the volumetric quantitative analysis of ascorbic acid and iron in dietary supplement tablets is presented. Powdered samples of the dietary supplement tablets were volumetrically titrated against electrolytically generated reagents, and the mass of dietary reagent in the tablet was determined from the…

  13. Automated potentiometric titrations in KCl/water-saturated octanol: method for quantifying factors influencing ion-pair partitioning.

    PubMed

    Scherrer, Robert A; Donovan, Stephen F

    2009-04-01

    The knowledge base of factors influencing ion pair partitioning is very sparse, primarily because of the difficulty in determining accurate log P(I) values of desirable low molecular weight (MW) reference compounds. We have developed a potentiometric titration procedure in KCl/water-saturated octanol that provides a link to log P(I) through the thermodynamic cycle of ionization and partitioning. These titrations have the advantage of being independent of the magnitude of log P, while maintaining a reproducibility of a few hundredths of a log P in the calculated difference between log P neutral and log P ion pair (diff (log P(N - I))). Simple model compounds can be used. The titration procedure is described in detail, along with a program for calculating pK(a)'' values incorporating the ionization of water in octanol. Hydrogen bonding and steric factors have a greater influence on ion pairs than they do on neutral species, yet these factors are missing from current programs used to calculate log P(I) and log D. In contrast to the common assumption that diff (log P(N - I)) is the same for all amines, they can actually vary more than 3 log units, as in our examples. A major factor affecting log P(I) is the ability of water and the counterion to approach the charge center. Bulky substituents near the charge center have a negative influence on log P(I). On the other hand, hydrogen bonding groups near the charge center have the opposite effect by lowering the free energy of the ion pair. The use of this titration method to determine substituent ion pair stabilization values (IPS) should bring about more accurate log D calculations and encourage species-specific QSAR involving log D(N) and log D(I). This work also brings attention to the fascinating world of nature's highly stabilized ion pairs.

  14. Microscale Titration in Schools Titration Competition.

    ERIC Educational Resources Information Center

    Clark, Michael J.

    1998-01-01

    Reviews the requirements of the National Titration Competition and describes how a team in a local competition used the technique. Compares microscale titration to conventional titration. Outlines the benefits of employing microscale techniques. (DDR)

  15. Competitive virus assay method for titration of noncytopathogenic bovine viral diarrhea viruses (END⁺ and END⁻ viruses).

    PubMed

    Muhsen, Mahmod; Ohi, Kota; Aoki, Hiroshi; Ikeda, Hidetoshi; Fukusho, Akio

    2013-03-01

    A new, reliable and secure virus assay method, named the competitive virus assay (CVA) method, has been established for the titration of bovine viral diarrhea viruses (BVDVs) that either show the exaltation of Newcastle disease virus (END) phenomenon or heterologous interference phenomenon (but not the END phenomenon). This method is based on the principle of (1) homologous interference between BVDVs, by using BVDV RK13/E(-) or BVDV RK13/E(+) strains as competitor virus, and (2) END phenomenon and heterologous interference, by using attenuated Newcastle disease virus (NDV) TCND strain as challenge virus. In titration of BVDV END(+) and BVDV END(-) viruses, no significant difference in estimated virus titer was observed between CVA and conventional methods. CVA method demonstrated comparable levels of sensitivity and accuracy as conventional END and interference methods, which require the use of a velogenic Miyadera strain of NDV and vesicular stomatitis virus (VSV), both of which are agents of high-risk diseases. As such, the CVA method is a safer alternative, with increased bio-safety and bio-containment, through avoidance of virulent strains that are commonly employed with conventional methods.

  16. Kinetic characteristics of polygalacturonase enzymes hydrolyzing galacturonic acid oligomers using isothermal titration calorimetry

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Polygalacturonase enzymes hydrolyze the polygalacturonic acid chains found in pectin. Interest in polygalacturonase enzymes continues as they are useful in a number of industrial processes and conversely, detrimental, as they are involved in maceration of economically important crops. While a good...

  17. Comparison of methods for isolation and titration of Crimean-Congo hemorrhagic fever virus.

    PubMed

    Shepherd, A J; Swanepoel, R; Leman, P A; Shepherd, S P

    1986-10-01

    The fluorescence focus assay and the plaque assay in CER cells were compared with mouse inoculation for the isolation and titration of Crimean-Congo hemorrhagic fever virus. The fluorescence focus assay and the plaque assay were of similar sensitivity, but both produced 10- to 100-fold lower titers than did mouse inoculation. For specimens from 26 Crimean-Congo hemorrhagic fever patients in South Africa, virus was isolated from 20 by mouse inoculation and from only 11 by cell culturing. Although cell cultures were less sensitive for the isolation of virus from clinical specimens, they produced diagnostic results much more rapidly.

  18. Molecular Characterization of the Gas-Particle Interface of Soot Sampled from a Diesel Engine Using a Titration Method.

    PubMed

    Tapia, A; Salgado, M S; Martín, María Pilar; Lapuerta, M; Rodríguez-Fernández, J; Rossi, M J; Cabañas, B

    2016-03-15

    Surface functional groups of two different types of combustion aerosols, a conventional diesel (EN 590) and a hydrotreated vegetable oil (HVO) soot, have been investigated using heterogeneous chemistry (i.e., gas-particle surface reactions). A commercial sample of amorphous carbon (Printex XE2-B) was analyzed as a reference substrate. A Knudsen flow reactor was used to carry out the experiments under molecular flow conditions. The selected gases for the titration experiments were: N(CH3)3 for the identification of acidic sites, NH2OH for the presence of carbonyl groups, CF3COOH and HCl for basic sites of different strength, and O3 and NO2 for reducing groups. Reactivity with N(CH3)3 indicates a lower density of acidic functionalities for Printex XE2-B in relation to diesel and HVO soot. Results for NH2OH experiments indicates that commercial amorphous carbon exhibits a lower abundance of available carbonyl groups at the interface compared to the results from diesel and HVO soot, the latter being the one with the largest abundance of carbonyl functions. Reactions with acids indicate the presence of weak basic oxides on the particle surface that preferentially interact with the strong acid CF3COOH. Finally, reactions with O3 and NO2 reveal that diesel and especially HVO have a significantly higher reactivity with both oxidizers compared to that of Printex XE2-B because they have more reducing sites by roughly a factor of 10 and 30, respectively. The kinetics of titration reactions have also been investigated.

  19. Molecular Characterization of the Gas-Particle Interface of Soot Sampled from a Diesel Engine Using a Titration Method.

    PubMed

    Tapia, A; Salgado, M S; Martín, María Pilar; Lapuerta, M; Rodríguez-Fernández, J; Rossi, M J; Cabañas, B

    2016-03-15

    Surface functional groups of two different types of combustion aerosols, a conventional diesel (EN 590) and a hydrotreated vegetable oil (HVO) soot, have been investigated using heterogeneous chemistry (i.e., gas-particle surface reactions). A commercial sample of amorphous carbon (Printex XE2-B) was analyzed as a reference substrate. A Knudsen flow reactor was used to carry out the experiments under molecular flow conditions. The selected gases for the titration experiments were: N(CH3)3 for the identification of acidic sites, NH2OH for the presence of carbonyl groups, CF3COOH and HCl for basic sites of different strength, and O3 and NO2 for reducing groups. Reactivity with N(CH3)3 indicates a lower density of acidic functionalities for Printex XE2-B in relation to diesel and HVO soot. Results for NH2OH experiments indicates that commercial amorphous carbon exhibits a lower abundance of available carbonyl groups at the interface compared to the results from diesel and HVO soot, the latter being the one with the largest abundance of carbonyl functions. Reactions with acids indicate the presence of weak basic oxides on the particle surface that preferentially interact with the strong acid CF3COOH. Finally, reactions with O3 and NO2 reveal that diesel and especially HVO have a significantly higher reactivity with both oxidizers compared to that of Printex XE2-B because they have more reducing sites by roughly a factor of 10 and 30, respectively. The kinetics of titration reactions have also been investigated. PMID:26886850

  20. Surfactant/detergent titration analysis method and apparatus for machine working fluids, surfactant-containing wastewater and the like

    DOEpatents

    Smith, Douglas D.; Hiller, John M.

    1998-01-01

    The present invention is an improved method and related apparatus for quantitatively analyzing machine working fluids and other aqueous compositions such as wastewater which contain various mixtures of cationic, neutral, and/or anionic surfactants, soluble soaps, and the like. The method utilizes a single-phase, non-aqueous, reactive titration composition containing water insoluble bismuth nitrate dissolved in glycerol for the titration reactant. The chemical reaction of the bismuth ion and glycerol with the surfactant in the test solutions results in formation of micelles, changes in micelle size, and the formation of insoluble bismuth soaps. These soaps are quantified by physical and chemical changes in the aqueous test solution. Both classical potentiometric analysis and turbidity measurements have been used as sensing techniques to determine the quantity of surfactant present in test solutions. This method is amenable to the analysis of various types of new, in-use, dirty or decomposed surfactants and detergents. It is a quick and efficient method utilizing a single-phase reaction without needing a separate extraction from the aqueous solution. It is adaptable to automated control with simple and reliable sensing methods. The method is applicable to a variety of compositions with concentrations from about 1% to about 10% weight. It is also applicable to the analysis of waste water containing surfactants with appropriate pre-treatments for concentration.

  1. Surfactant/detergent titration analysis method and apparatus for machine working fluids, surfactant-containing wastewater and the like

    DOEpatents

    Smith, D.D.; Hiller, J.M.

    1998-02-24

    The present invention is an improved method and related apparatus for quantitatively analyzing machine working fluids and other aqueous compositions such as wastewater which contain various mixtures of cationic, neutral, and/or anionic surfactants, soluble soaps, and the like. The method utilizes a single-phase, non-aqueous, reactive titration composition containing water insoluble bismuth nitrate dissolved in glycerol for the titration reactant. The chemical reaction of the bismuth ion and glycerol with the surfactant in the test solutions results in formation of micelles, changes in micelle size, and the formation of insoluble bismuth soaps. These soaps are quantified by physical and chemical changes in the aqueous test solution. Both classical potentiometric analysis and turbidity measurements have been used as sensing techniques to determine the quantity of surfactant present in test solutions. This method is amenable to the analysis of various types of new, in-use, dirty or decomposed surfactants and detergents. It is a quick and efficient method utilizing a single-phase reaction without needing a separate extraction from the aqueous solution. It is adaptable to automated control with simple and reliable sensing methods. The method is applicable to a variety of compositions with concentrations from about 1% to about 10% weight. It is also applicable to the analysis of waste water containing surfactants with appropriate pre-treatments for concentration. 1 fig.

  2. Studies on Newcastle disease virus; an evaluation of the method of titration.

    PubMed

    BANG, F B

    1948-08-01

    The application of the 50 per cent embryo mortality to a study of the virus of Newcastle is described. It has been evaluated by a series of duplicate titrations of the same sample of virus. In seven such titrations the largest difference between the two was 10(-0.4). It is therefore believed that a difference of 0.6 log is probably significant and of 1.0 log almost certainly significant. This would mean that we can almost certainly detect a loss of 90 per cent of activity. Neither temperature of incubation nor route of inoculation in the test embryos had consistent effect on the measurement of virus activity. The effect of increasing age of the incubated embryo, from 10 days up to 16 days, is slight and inconsistent. The addition of chicken red blood cells to a dilution of virus may lower the titer of the preparation, but the change is not sufficient to be of importance in the routine handling of the virus.

  3. Universal tracer monitored titrations.

    PubMed

    DeGrandpre, Michael D; Martz, Todd R; Hart, Robert D; Elison, David M; Zhang, Alice; Bahnson, Anna G

    2011-12-15

    Titrations, while primarily known as the chemical rite of passage for fledgling science students, are still widely used for chemical analysis. With its many years of existence and improvement, the method would seem an unlikely candidate for innovation, yet it is desirable, in this age of autonomous sensing where analyzers may be sent into space or to the bottom of the ocean, to have a simplified titrimetric method that does not rely upon volumetric or gravimetric measurement of sample and titrant. In previous work on the measurement of seawater alkalinity, we found that use of a tracer in the titrant eliminates the need to measure mass or volume. Here, we show the versatility of the method for diverse types of titrations and tracers. The results suggest that tracers may be employed in all types of titrations, opening the door for greatly simplified laboratory and field-based chemical analysis.

  4. Modeling Precipitation and Sorption of Al, U and Co-contaminants during Titration of Acidic Sediments in Recirculation Flow-Through Experiments

    SciTech Connect

    Tang, Guoping; Luo, Wensui; Brooks, Scott C; Watson, David B; Gu, Baohua

    2013-01-01

    We conducted batch and recirculating column titration tests with contaminated acidic sediments with controlled CO2 in the headspace, and extended the geochemical model by Gu et al. (2003, GCA) to better understand and quantify the reactions governing trace metal fate in the subsurface. The sediment titration curve showed slow pH increase due to strong buffering by Al precipitation and CO2 uptake. Assuming precipitation of basaluminite at low saturation index (SI=-4), and decreasing cation exchange selectivity coefficient (kNa\\Al=0.3), the predictions are close to the observed pH and Al; and the model explains 1) the observed Ca, Mg, and Mn concentration decrease by cation exchange with sorbed Al, and 2) the decrease of U by surface complexation with Fe hydroxides at low pH, and precipitation as liebigite (Ca2UO2(CO3)3:10H2O) at pH>5.5. Without further adjustment geochemical parameters, the model describes reasonably well previous sediment and column titration tests without CO2 in the headspace, as well as the new large column test. The apparent inhibition of U and Ni decrease in the large column can be explained by formation of aqueous carbonate complexes and/or competition with carbonate for surface sites. These results indicated that ignoring labile solid phase Al would underestimate base requirement in titration of acidic aquifers.

  5. Amperometric, Bipotentiometric, and Coulometric Titration.

    ERIC Educational Resources Information Center

    Stock, John T.

    1980-01-01

    Discusses recent review articles in various kinds of titration. Also discusses new research in apparatus and methodology, acid-base reactions, precipitation and complexing reactions, oxidation-reduction reactions, and nomenclature. Cites 338 references. (CS)

  6. Colorblindness and Titrations with Visual Indicators.

    ERIC Educational Resources Information Center

    Diehl, Harvey; And Others

    1985-01-01

    Discusses various issues related to colorblind students performing titrations with visual indicators. Includes tables showing precisions in the titration of a weak acid by colorblind students using phenolphthalein and thymolphthalein and in the titration of a weak base by colorblind persons using methyl red and bromcresol green. (JN)

  7. The use of graphite electrodes in potentiometric titrations

    SciTech Connect

    Selig, W.S.

    1987-04-01

    The use of various types of graphite as indicator electrodes in potentiometry has been limited to acid-base and redox titrations. We have expanded the range of feasible titrations to: (1) precipitation titrations; (2) acid-base titrations; (3) compleximetric titrations; and (4) redox titrations. Graphite covered with an organic membrane containing poly(vinyl chloride) (PVC) and a plasticizer is particularly useful in monitoring the endpoints of titrations in which insoluble ion-pairs are formed. The potentiometric titration of fluoride vs La(III) or Th(IV), or of sulfate vs Pb (II) or Ba(II), which can be monitored with a plain carbon rod, is discussed.

  8. Comparison of Densitometry, Refractive Index, and Classical Permanganate Titration Assay Methods with Commercially Available Rocket-Grade Hydrogen Peroxide

    NASA Astrophysics Data System (ADS)

    McPherson, M. D.

    2004-10-01

    Using classical methods of assay analysis for highly concentrated rocket grade hydrogen peroxide (RGHP, H2O2), comparison and correlation to historical values using currently available commercial-source RGHP is presented. In particular, fluid densities, refractive index, and the potassium permanganate (KMnO4) wet chemical method of analysis are compared. Discussion includes those analytical processes that support field use, with attendant rapid, accurate, and verifiable concentration determinations of RGHP. Method accuracy is ranked as Manual KMnO4 Titration > Refractive Index > Density, based on vendor certifications, with the refractive index method having best precision (+/-0.02%wt H2O2). Results from densitometry data are anomalous, with variability likely due to less mature measurement techniques used in this study.

  9. Investigation of the ionic strength dependence of Ulva lactuca acid functional group pK(a)s by manual alkalimetric titrations.

    PubMed

    Schijf, Johan; Ebling, Alina M

    2010-03-01

    We performed a series of manual alkalimetric titrations in NaCl solutions (0.01-5.0 M) at T = 25 degrees C on both fresh and dehydrated samples of the marine chlorophyte Ulva lactuca (sea lettuce), a strong metal accumulator holding considerable promise in biosorbent and biomonitor applications. Functional groups were characterized in terms of their number, site densities, and acid dissociation constants (pK(a)s). FITEQL4.0 modeling shows that, at any ionic strength, titration curves for dehydrated biomass in the pH range 2-10 are adequately described by three functional groups with remarkably uniform site densities of about 5 x 10(-4) mol/g. Lower site densities for fresh U. lactuca are consistent with approximately 87% water content. The pK(a)s display pronounced ionic strength dependent behavior obeying an extended Debye-Huckel relation. Extrapolation to I = 0 yields values of 4.26 +/- 0.04, 6.44 +/- 0.02, and 9.56 +/- 0.04. This information by itself is insufficient to unambiguously identify the groups. Similar site densities suggest that all three are linked to major molecular building blocks of the cell material, pointing to carboxylic acids, phosphate esters, and amines as likely candidates. Highly acidic sulfate esters, not detected in our titrations, may also play a role in trace metal adsorption on U. lactuca.

  10. Acid-base properties of humic substances from composted and thermally-dried sewage sludges and amended soils as determined by potentiometric titration and the NICA-Donnan model.

    PubMed

    Fernández, José M; Plaza, César; Senesi, Nicola; Polo, Alfredo

    2007-09-01

    The acid-base properties of humic acids (HAs) and fulvic acids (FAs) isolated from composted sewage sludge (CS), thermally-dried sewage sludge (TS), soils amended with either CS or TS at a rate of 80 t ha(-1)y(-1) for 3y and the corresponding unamended soil were investigated by use of potentiometric titrations. The non-ideal competitive adsorption (NICA)-Donnan model for a bimodal distribution of proton binding sites was fitted to titration data by use of a least-squares minimization method. The main fitting parameters of the NICA-Donnan model obtained for each HA and FA sample included site densities, median affinity constants and widths of affinity distributions for proton binding to low and high affinity sites, which were assumed to be, respectively, carboxylic- and phenolic-type groups. With respect to unamended soil HA and FA, the HAs and FAs from CS, and especially TS, were characterized by smaller acidic functional group contents, larger proton binding affinities of both carboxylic- and phenolic-type groups, and smaller heterogeneity of carboxylic and phenolic-type groups. Amendment with CS or TS led to a decrease of acidic functional group contents and a slight increase of proton binding affinities of carboxylic- and phenolic-type groups of soil HAs and FAs. These effects were more evident in the HA and FA fractions from CS-amended soil than in those from TS-amended soil.

  11. Interactions between poly(acrylic acid) and sodium dodecyl sulfate: isothermal titration calorimetric and surfactant ion-selective electrode studies.

    PubMed

    Wang, C; Tam, K C

    2005-03-24

    Interaction between a monodispersed poly(acrylic acid) (PAA) (M(W) = 5670 g/mol, M(w)/M(n) = 1.02) with sodium dodecyl sulfate (SDS) was investigated using isothermal titration calorimetry (ITC), ion-selective electrode (ISE), and dynamic light scattering measurements. Contrary to previous studies, we report for the first time evidence of interaction between SDS and PAA when the degree of neutralization (alpha) of PAA is lower than 0.2. Hydrocarbon chains of SDS cooperatively bind to apolar segments of PAA driven by hydrophobic interaction. The interaction is both enthalpy and entropy favored (deltaH is negative but deltaS is positive). In 0.05 wt % PAA solution, the SDS concentration corresponding to the onset of binding (i.e., CAC) is approximately 2.4 mM and the saturation concentration (i.e., C(S)) is approximately 13.3 mM when alpha = 0. When PAA was neutralized and ionized, the binding was hindered by the enhanced electrostatic repulsion between negatively charged SDS and PAA chains and improved solubility of the polymer. With increasing alpha to 0.2, CAC increases to approximately 6.2 mM, C(S) drops to 8.6 mM, and the interaction is significantly weakened where the amount of bound SDS on PAA is reduced considerably. The values of CAC and C(S) derived from different techniques are in good agreement. The binding results in the formation of mixed micelles on apolar PAA coils, which then expands and dissociates into single PAA chains. The majority of unneutralized PAA molecules exist as single polymer chains stabilized by bound SDS micelles in solution after the saturation concentration.

  12. Microbiological profiles, pH, and titratable acidity of chorizo and salchichón (two Spanish dry fermented sausages) manufactured with ostrich, deer, or pork meat.

    PubMed

    Capita, Rosa; Llorente-Marigómez, Sandra; Prieto, Miguel; Alonso-Calleja, Carlos

    2006-05-01

    Microbial counts, pH, and titratable acidity were determined in 102 Spanish dry fermented sausages (chorizo and salchichón) made with ostrich, deer, or pork meat. Average microbial counts (log CFU per gram) varied from 5.46 +/- 0.24 to 8.25 +/- 0.80 (total viable counts), from 4.79 +/- 0.36 to 7.99 +/- 0.20 (psychrotrophs), from 0.00 +/- 0.00 to 0.99 +/- 1.10 (undetectable values were assumed to be zero) (Enterobacteriaceae), from 0.00 +/- 0.00 to 4.27 +/- 1.47 (enterococci), from 5.15 +/- 1.15 to 8.46 +/- 0.49 (lactic acid bacteria), from 3.08 +/- 0.44 to 6.59 +/- 1.76 (Micrococcaceae), from 2.27 +/- 1.53 to 5.11 +/- 1.81 (molds and yeasts), from 0.00 +/- 0.00 to 2.25 +/- 0.81 (pseudomonads), and from 0.00 +/- 0.00 to 2.78 +/- 0.46 (Brochothrix thermosphacta). Average pH and titratable acidity varied from 5.07 +/- 0.25 to 5.63 +/- 0.51 (pH units) and from 0.30 +/- 0.01 to 0.86 +/- 0.19 (% lactic acid). Both type of sausage (P < 0.05) and species of meat (P < 0.001) influenced microbial counts. Salchich6n samples showed lower average values than chorizo samples for most microbial groups (significant for Enterobacteriaceae, lactic acid bacteria, and B. thermosphacta) and titratable acidity. Sausages made from pork showed the highest microbial loads for total viable counts, psychrotrophs, Enterobacteriaceae, enterococci, lactic acid bacteria, and yeasts and molds. Higher counts were observed only for pseudomonads in ostrich sausages. B. thermosphacta levels were similar for all species of meat. The highest average pH value was observed in sausages made from ostrich meat, and the lowest titratable acidity level was found in pork sausages.

  13. Tracer monitored titrations: measurement of total alkalinity.

    PubMed

    Martz, Todd R; Dickson, Andrew G; DeGrandpre, Michael D

    2006-03-15

    We introduce a new titration methodology, tracer monitored titration (TMT), in which analyses are free of volumetric and gravimetric measurements and insensitive to pump precision and reproducibility. Spectrophotometric monitoring of titrant dilution, rather than volume increment, lays the burden of analytical performance solely on the spectrophotometer. In the method described here, the titrant is a standardized mixture of acid-base indicator and strong acid. Dilution of a pulse of titrant in a titration vessel is tracked using the total indicator concentration measured spectrophotometrically. The concentrations of reacted and unreacted indicator species, derived from Beer's law, are used to calculate the relative proportions of titrant and sample in addition to the equilibrium position (pH) of the titration mixture. Because the method does not require volumetric or gravimetric additions of titrant, simple low-precision pumps can be used. Here, we demonstrate application of TMT for analysis of total alkalinity (A(T)). High-precision, high-accuracy seawater A(T) measurements are crucial for understanding, for example, the marine CaCO3 budget and saturation state, anthropogenic CO2 penetration into the oceans, calcareous phytoplankton blooms, and coral reef dynamics. We present data from 286 titrations on three types of total alkalinity standards: Na2CO3 in 0.7 mol kg x soln(-1) NaCl, NaOH in 0.7 mol kg x soln(-1) NaCl, and a seawater Certified Reference Material (CRM). Based on Na2CO3 standards, the accuracy and precision are +/-0.2 and +/-0.1% (4 and 2 micromol kg x soln(-1) for A(T) approximately 2100-2500 micromol kg x soln(-1), n = 242), using low-precision solenoid pumps to introduce sample and titrant. Similar accuracy and precision were found for analyses run 42 days after the initial experiments. Excellent performance is achieved by optimizing the spectrophotometric detection system and relying upon basic chemical thermodynamics for calculating the

  14. Calcium-buffering effects of gluconate and nucleotides, as determined by a novel fluorimetric titration method

    PubMed Central

    Woehler, Andrew; Lin, Kun-Han; Neher, Erwin

    2014-01-01

    Significantly more Ca2+ influx is required for eliciting release of neurotransmitter during whole cell patch clamp recording in the Calyx of Held, when gluconate with 3 mm free ATP is used as pipette filling solution, as compared to a methanesulfonate-based solution with excess Mg2+. This reduction in efficiency of Ca2+ in eliciting release is due to low-affinity Ca2+ binding of both gluconate and ATP2− anions. To study these effects we developed a simple fluorimeteric titration procedure, which reports the dissociation constant, KD, of a given Ca2+ indicator dye, multiplied by 1 plus the sum of Ca2+ binding ratios of any anions, which act as low-affinity Ca2+ ligands. For solutions without Ca2+ binding anions we find KD values for Fura2FF ranging from 11.5 ± 1.7 to 15.6 ± 7.47 μm depending on the dominant anion used. For Fura6F and KCl-based solutions we find KD = 17.8 ± 1.3 μm. For solutions with gluconate as the main anion and for solutions that contain nucleotides, such as ATP and GTP, we find much higher values for the product. Assuming that the KD of the indicator dye is equal to that of KCl-based solutions we calculate the summed Ca2+ binding ratios and find a value of 3.55 for a solution containing 100 mm potassium gluconate and 4 mm ATP. Gluconate contributes a value of 1.75 to this number, while the contribution of ATP depends strongly on the presence of Mg2+ and varies from 0.8 (with excess Mg2+) to 13.8 (in the presence of 3 mm free ATP). Methanesulfonate has negligible Ca2+ binding capacity. These results explain the reduced efficiency of Ca2+ influx in the presence of gluconate or nucleotides, as these anions are expected to intercept Ca2+ ions at short distance. PMID:25194050

  15. Iodine losses during Winkler titrations

    NASA Astrophysics Data System (ADS)

    Knapp, George P.; Stalcup, Marvel C.; Stanley, Robert J.

    1991-01-01

    An experiment designed to measure iodine loss during the aliquot version of the Winkler titration for dissolved oxygen in seawater shows that 0.01-0.03 ml l -1 equivalent oxygen is lost at typical oceanic concentrations in the method presently used. A standardization technique, which mimics that employed during the titration of seawater samples, compensates for this iodine loss throughout the oceanic range. This result, contradicting an earlier report by GREEN and CARRITT (1966, Analyst, 91, 207-208), demonstrates that the whole-bottle method of oxygen titration is not to be preferred over the aliquot method.

  16. Method for isolating nucleic acids

    SciTech Connect

    Hurt, Jr., Richard Ashley; Elias, Dwayne A.

    2015-09-29

    The current disclosure provides methods and kits for isolating nucleic acid from an environmental sample. The current methods and compositions further provide methods for isolating nucleic acids by reducing adsorption of nucleic acids by charged ions and particles within an environmental sample. The methods of the current disclosure provide methods for isolating nucleic acids by releasing adsorbed nucleic acids from charged particles during the nucleic acid isolation process. The current disclosure facilitates the isolation of nucleic acids of sufficient quality and quantity to enable one of ordinary skill in the art to utilize or analyze the isolated nucleic acids for a wide variety of applications including, sequencing or species population analysis.

  17. Tracer-monitored flow titrations.

    PubMed

    Sasaki, Milton K; Rocha, Diogo L; Rocha, Fábio R P; Zagatto, Elias A G

    2016-01-01

    The feasibility of implementing tracer-monitored titrations in a flow system is demonstrated. A dye tracer is used to estimate the instant sample and titrant volumetric fractions without the need for volume, mass or peak width measurements. The approach was applied to spectrophotometric flow titrations involving variations of sample and titrant flow-rates (i.e. triangle programmed technique) or concentration gradients established along the sample zone (i.e. flow injection system). Both strategies required simultaneous monitoring of two absorbing species, namely the titration indicator and the dye tracer. Mixing conditions were improved by placing a chamber with mechanical stirring in the analytical path aiming at to minimize diffusional effects. Unlike most of flow-based titrations, the innovation is considered as a true titration, as it does not require a calibration curve thus complying with IUPAC definition. As an application, acidity evaluation in vinegars involving titration with sodium hydroxide was selected. Phenolphthalein and brilliant blue FCF were used as indicator and dye tracer, respectively. Effects of sample volume, titrand/titrant concentrations and flow rates were investigated aiming at improved accuracy and precision. Results were reliable and in agreement with those obtained by a reference titration procedure.

  18. Tracer-monitored flow titrations.

    PubMed

    Sasaki, Milton K; Rocha, Diogo L; Rocha, Fábio R P; Zagatto, Elias A G

    2016-01-01

    The feasibility of implementing tracer-monitored titrations in a flow system is demonstrated. A dye tracer is used to estimate the instant sample and titrant volumetric fractions without the need for volume, mass or peak width measurements. The approach was applied to spectrophotometric flow titrations involving variations of sample and titrant flow-rates (i.e. triangle programmed technique) or concentration gradients established along the sample zone (i.e. flow injection system). Both strategies required simultaneous monitoring of two absorbing species, namely the titration indicator and the dye tracer. Mixing conditions were improved by placing a chamber with mechanical stirring in the analytical path aiming at to minimize diffusional effects. Unlike most of flow-based titrations, the innovation is considered as a true titration, as it does not require a calibration curve thus complying with IUPAC definition. As an application, acidity evaluation in vinegars involving titration with sodium hydroxide was selected. Phenolphthalein and brilliant blue FCF were used as indicator and dye tracer, respectively. Effects of sample volume, titrand/titrant concentrations and flow rates were investigated aiming at improved accuracy and precision. Results were reliable and in agreement with those obtained by a reference titration procedure. PMID:26703261

  19. An arbitrary correction function for CO(2) evolution in acid-base titrations and its use in multiparametric refinement of data.

    PubMed

    Wozniak, M; Nowogrocki, G

    1981-08-01

    A great number of acid-base titrations are performed under an inert gas flow: in the procedure, a variable amount of CO(2)-from carbonated reactants-is carried away and thus prevents strict application of mass-balance equations. A function for the CO(2) evolution is proposed and introduced into the general expression for the volume of titrant. Use of this expression in multiparametric refinement yields, besides the usual values (concentrations, acidity constants...), a parameter characteristic of this departure of CO(2). Furthermore, a modified weighting factor is introduced to take into account the departure from equilibrium caused by the slow CO(2) evolution. The validity of these functions was successfully tested on three typical examples: neutralization of strong acid by sodium carbonate, of sodium carbonate by strong acid, and of a mixture of hydrochloric acid, 4-nitrophenol and phenol by carbonated potassium hydroxide.

  20. Analysis of aqueous solutions by near-infrared spectrometry (NIRS). I. Titrations of strong acids with strong bases

    NASA Astrophysics Data System (ADS)

    Molt, K.; Cho, Y. J.

    1995-04-01

    The titration of 1 M HCl with 2 M NaOH was observed by NIR-spectrometry. The measured spectra can be evaluated quantitatively for calibration of the H +- and OH --concentration. The NIRS errors with respect to the prediction of the pH and the equivalence point are discussed.

  1. Responses of grape berry anthocyanin and titratable acidity to the projected climate change across the Western Australian wine regions

    NASA Astrophysics Data System (ADS)

    Barnuud, Nyamdorj N.; Zerihun, Ayalsew; Mpelasoka, Freddie; Gibberd, Mark; Bates, Bryson

    2014-08-01

    More than a century of observations has established that climate influences grape berry composition. Accordingly, the projected global climate change is expected to impact on grape berry composition although the magnitude and direction of impact at regional and subregional scales are not fully known. The aim of this study was to assess potential impacts of climate change on levels of berry anthocyanin and titratable acidity (TA) of the major grapevine varieties grown across all of the Western Australian (WA) wine regions. Grape berry anthocyanin and TA responses across all WA wine regions were projected for 2030, 2050 and 2070 by utilising empirical models that link these berry attributes and climate data downscaled (to ˜5 km resolution) from the csiro_mk3_5 and miroc3_2_medres global climate model outputs under IPCC SRES A2 emissions scenario. Due to the dependence of berry composition on maturity, climate impacts on anthocyanin and TA levels were assessed at a common maturity of 22 °Brix total soluble solids (TSS), which necessitated the determination of when this maturity will be reached for each variety, region and warming scenario, and future period. The results indicate that both anthocyanin and TA levels will be affected negatively by a warming climate, but the magnitude of the impacts will differ between varieties and wine regions. Compared to 1990 levels, median anthocyanins concentrations are projected to decrease, depending on global climate model, by up to 3-12 % and 9-33 % for the northern wine regions by 2030 and 2070, respectively while 2-18 % reductions are projected in the southern wine regions for the same time periods. Patterns of reductions in the median Shiraz berry anthocyanin concentrations are similar to that of Cabernet Sauvignon; however, the magnitude is lower (up to 9-18 % in southern and northern wine regions respectively by 2070). Similarly, uneven declines in TA levels are projected across the study regions. The largest reductions

  2. Responses of grape berry anthocyanin and titratable acidity to the projected climate change across the Western Australian wine regions.

    PubMed

    Barnuud, Nyamdorj N; Zerihun, Ayalsew; Mpelasoka, Freddie; Gibberd, Mark; Bates, Bryson

    2014-08-01

    More than a century of observations has established that climate influences grape berry composition. Accordingly, the projected global climate change is expected to impact on grape berry composition although the magnitude and direction of impact at regional and subregional scales are not fully known. The aim of this study was to assess potential impacts of climate change on levels of berry anthocyanin and titratable acidity (TA) of the major grapevine varieties grown across all of the Western Australian (WA) wine regions. Grape berry anthocyanin and TA responses across all WA wine regions were projected for 2030, 2050 and 2070 by utilising empirical models that link these berry attributes and climate data downscaled (to ∼5 km resolution) from the csiro_mk3_5 and miroc3_2_medres global climate model outputs under IPCC SRES A2 emissions scenario. Due to the dependence of berry composition on maturity, climate impacts on anthocyanin and TA levels were assessed at a common maturity of 22 °Brix total soluble solids (TSS), which necessitated the determination of when this maturity will be reached for each variety, region and warming scenario, and future period. The results indicate that both anthocyanin and TA levels will be affected negatively by a warming climate, but the magnitude of the impacts will differ between varieties and wine regions. Compared to 1990 levels, median anthocyanins concentrations are projected to decrease, depending on global climate model, by up to 3-12 % and 9-33 % for the northern wine regions by 2030 and 2070, respectively while 2-18 % reductions are projected in the southern wine regions for the same time periods. Patterns of reductions in the median Shiraz berry anthocyanin concentrations are similar to that of Cabernet Sauvignon; however, the magnitude is lower (up to 9-18 % in southern and northern wine regions respectively by 2070). Similarly, uneven declines in TA levels are projected across the study regions. The largest reductions

  3. Development and optimization of a real-time quantitative PCR-based method for the titration of AAV-2 vector stocks.

    PubMed

    Veldwijk, Marlon R; Topaly, Julian; Laufs, Stephanie; Hengge, Ulrich R; Wenz, Frederik; Zeller, W Jens; Fruehauf, Stefan

    2002-08-01

    Despite the clinical application of adeno-associated virus (AAV) gene therapy, the titration of viral stocks has not yet been standardized. This complicates the comparison of viral stocks between laboratories. Functional titering of AAV is time-consuming, requires the manipulation of hazardous material, and often has a high degree of variability. We established an optimized real-time quantitative polymerase chain reaction (RQ-PCR) titration assay to determine viral titers and compared it with a functional green fluorescent protein (GFP)-based titration method. With a combination of improved lysis procedures and RQ-PCR protocols we could decrease the intraexperimental coefficient of variation (CV) from 0.24 +/- 0.03 to 0.042 +/- 0.004 and the interexperimental CV from 0.34 +/- 0.06 to 0.093 +/- 0.028 following functional and RQPCR-based titration, respectively. This low variability conforms to even the strictest quality standards required, for example, in clinical laboratories. The highly standardized titration by RQPCR described here will be especially advantageous for groups working on AAV-based gene therapy in a good manufacturing practice setting.

  4. Nucleic acid detection methods

    DOEpatents

    Smith, C.L.; Yaar, R.; Szafranski, P.; Cantor, C.R.

    1998-05-19

    The invention relates to methods for rapidly determining the sequence and/or length a target sequence. The target sequence may be a series of known or unknown repeat sequences which are hybridized to an array of probes. The hybridized array is digested with a single-strand nuclease and free 3{prime}-hydroxyl groups extended with a nucleic acid polymerase. Nuclease cleaved heteroduplexes can be easily distinguish from nuclease uncleaved heteroduplexes by differential labeling. Probes and target can be differentially labeled with detectable labels. Matched target can be detected by cleaving resulting loops from the hybridized target and creating free 3-hydroxyl groups. These groups are recognized and extended by polymerases added into the reaction system which also adds or releases one label into solution. Analysis of the resulting products using either solid phase or solution. These methods can be used to detect characteristic nucleic acid sequences, to determine target sequence and to screen for genetic defects and disorders. Assays can be conducted on solid surfaces allowing for multiple reactions to be conducted in parallel and, if desired, automated. 18 figs.

  5. Nucleic Acid Detection Methods

    DOEpatents

    Smith, Cassandra L.; Yaar, Ron; Szafranski, Przemyslaw; Cantor, Charles R.

    1998-05-19

    The invention relates to methods for rapidly determining the sequence and/or length a target sequence. The target sequence may be a series of known or unknown repeat sequences which are hybridized to an array of probes. The hybridized array is digested with a single-strand nuclease and free 3'-hydroxyl groups extended with a nucleic acid polymerase. Nuclease cleaved heteroduplexes can be easily distinguish from nuclease uncleaved heteroduplexes by differential labeling. Probes and target can be differentially labeled with detectable labels. Matched target can be detected by cleaving resulting loops from the hybridized target and creating free 3-hydroxyl groups. These groups are recognized and extended by polymerases added into the reaction system which also adds or releases one label into solution. Analysis of the resulting products using either solid phase or solution. These methods can be used to detect characteristic nucleic acid sequences, to determine target sequence and to screen for genetic defects and disorders. Assays can be conducted on solid surfaces allowing for multiple reactions to be conducted in parallel and, if desired, automated.

  6. Geochemical modeling of reactions and partitioning of trace metals and radionuclides during titration of contaminated acidic sediments

    SciTech Connect

    Zhang, Fan; Parker, Jack C.; Brooks, Scott C; Watson, David B; Jardine, Philip M; Gu, Baohua

    2008-01-01

    This study investigated sorption of uranium and technetium onto aluminum and iron hydroxides during titration of a contaminated groundwater using both Na hydroxide and carbonate as titrants. The contaminated groundwater has a low pH of 3.8 and high concentrations of NO3-, SO42-, Al, Ca, Mg, Mn, trace metals such as Ni and Co, and radionuclides such as U and Tc. During titration, most Al and Fe were precipitated out at pH above ~4.5. U as well as Tc was found to be removed from aqueous phase at pH below ~5.5, but to some extent released at higher pH values. An earlier geochemical equilibrium reaction path model that considered aqueous complexation and precipitation/dissolution reactions predicted mineral precipitation and adequately described concentration variations of Al, Fe and some other metal cations, but failed to predict sulfate, U and Tc concentrations during titration. Previous studies have shown that Fe- and Al-oxyhydroxides strongly sorb dissolved sulfate, U and Tc species. Therefore, an anion exchange model was developed for the sorption of sulfate, U and Tc onto Al and Fe hydroxides. With the additional consideration of the anion exchange reactions, concentration profiles of sulfate, U and Tc were more accurately predicted. Results of this study indicate that consideration of complex reactions such as sorption/desorption on mixed mineral phases, in addition to hydrolysis and precipitation, could improve the prediction of various contaminants during pre- and post-groundwater treatment practices.

  7. A study of pH-dependent photodegradation of amiloride by a multivariate curve resolution approach to combined kinetic and acid-base titration UV data.

    PubMed

    De Luca, Michele; Ioele, Giuseppina; Mas, Sílvia; Tauler, Romà; Ragno, Gaetano

    2012-11-21

    Amiloride photostability at different pH values was studied in depth by applying Multivariate Curve Resolution Alternating Least Squares (MCR-ALS) to the UV spectrophotometric data from drug solutions exposed to stressing irradiation. Resolution of all degradation photoproducts was possible by simultaneous spectrophotometric analysis of kinetic photodegradation and acid-base titration experiments. Amiloride photodegradation showed to be strongly dependent on pH. Two hard modelling constraints were sequentially used in MCR-ALS for the unambiguous resolution of all the species involved in the photodegradation process. An amiloride acid-base system was defined by using the equilibrium constraint, and the photodegradation pathway was modelled taking into account the kinetic constraint. The simultaneous analysis of photodegradation and titration experiments revealed the presence of eight different species, which were differently distributed according to pH and time. Concentration profiles of all the species as well as their pure spectra were resolved and kinetic rate constants were estimated. The values of rate constants changed with pH and under alkaline conditions the degradation pathway and photoproducts also changed. These results were compared to those obtained by LC-MS analysis from drug photodegradation experiments. MS analysis allowed the identification of up to five species and showed the simultaneous presence of more than one acid-base equilibrium.

  8. [Comparison of 2 in vitro methods of titration of antibodies neutralizing the classic swine plague virus].

    PubMed

    Corthier, G

    1976-01-01

    Swine fever virus neutralization have been studied by the following method: variable plaque forming units numbers are mixed with a predetermined serum dilution. Infectivity is measured before (VO) and after (Vr) neutralization reaction. Neutralization index (N.I. = logVo/V) represent the difference between the two titers. It had been demonstrated that mass law is a good approximation to describe swine fever virus neutralization. So the most useful form in which to express the relation between N.I. and dilution logarithm (log D) is NI = NIo--K log D (I) where K is the constante corresponding to the slope of neutralization curve. A sligh K variation is observed according to immunoglobulin classes sharing antibody activity. Average K value is equal to 3. NIO is the NI obtained when extrapolating the curve to log D = o. These results have permitted to compare the two seroneutralization methods commonly used: "constant serum" method and "variable serum" method (determination of serum dilution inhibiting 50% of infectivity: D50). From formula (I) the following relation can be obtained: see formular. Theoretical and experimental log D50 values were appromaximatively the same (difference inferior or equal to 0.3). So D50 can be calculated by both methods.

  9. Isothermal titration calorimetry method for determination of cyclodextrin complexation thermodynamics between artemisinin and naproxen under varying environmental conditions.

    PubMed

    Illapakurthy, Ashok C; Wyandt, Christy M; Stodghill, Steven P

    2005-02-01

    A novel isothermal titration calorimetry method was used to determine the complexation thermodynamics for hydroxypropyl-beta-cyclodextrin with artemisinin and naproxen at varying temperature and pH. The new method is very useful for studying complexation reactions between cyclodextrin and drugs with poor solubility and all the thermodynamic parameters of the cyclodextrin complexation were determined. The analysis of the thermodynamic data reveals involvement of hydrophobic bonding in the cyclodextrin complexes studied. The data also reveals the presence of enthalpy-entropy compensation in the system and provide information as to the orientation of the drug molecule inside the cyclodextrin cavity. From the thermodynamic parameters for dissociation of HPBCD complexes of artemisinin and naproxen at pH 2 it is concluded that the complexation is primarily driven by enthalpy with entropic assistance at all temperatures studied. From the dissociation studies of HPBCD complexes of naproxen at pH 10 it is concluded that the complexation is predominantly driven by entropy and moderately by enthalpy at lower temperatures and by enthalpy with entropic assistance at higher temperatures. PMID:15661505

  10. Acid-base titrations of functional groups on the surface of the thermophilic bacterium Anoxybacillus flavithermus: comparing a chemical equilibrium model with ATR-IR spectroscopic data.

    PubMed

    Heinrich, Hannah T M; Bremer, Phil J; Daughney, Christopher J; McQuillan, A James

    2007-02-27

    Acid-base functional groups at the surface of Anoxybacillus flavithermus (AF) were assigned from the modeling of batch titration data of bacterial suspensions and compared with those determined from in situ infrared spectroscopic titration analysis. The computer program FITMOD was used to generate a two-site Donnan model (site 1: pKa = 3.26, wet concn = 2.46 x 10(-4) mol g(-1); site 2: pKa = 6.12, wet concn = 6.55 x 10(-5) mol g(-1)), which was able to describe data for whole exponential phase cells from both batch acid-base titrations at 0.01 M ionic strength and electrophoretic mobility measurements over a range of different pH values and ionic strengths. In agreement with information on the composition of bacterial cell walls and a considerable body of modeling literature, site 1 of the model was assigned to carboxyl groups, and site 2 was assigned to amino groups. pH difference IR spectra acquired by in situ attenuated total reflection infrared (ATR-IR) spectroscopy confirmed the presence of carboxyl groups. The spectra appear to show a carboxyl pKa in the 3.3-4.0 range. Further peaks were assigned to phosphodiester groups, which deprotonated at slightly lower pH. The presence of amino groups could not be confirmed or discounted by IR spectroscopy, but a positively charged group corresponding to site 2 was implicated by electrophoretic mobility data. Carboxyl group speciation over a pH range of 2.3-10.3 at two different ionic strengths was further compared to modeling predictions. While model predictions were strongly influenced by the ionic strength change, pH difference IR data showed no significant change. This meant that modeling predictions agreed reasonably well with the IR data for 0.5 M ionic strength but not for 0.01 M ionic strength.

  11. Comparison between antigen-capture ELISA and conventional methods used for titration of infectious bursal disease virus.

    PubMed

    Hassan, M K; Saif, Y M; Shawky, S

    1996-01-01

    Two antigen-capture enzyme-linked immunosorbent assays (polyclonal and monoclonal AC-ELISAs) were developed and evaluated for titration of infectious bursal disease viruses (IBDVs) propagated in different host systems, namely BGM-70 continuous cell line, primary chicken embryo fibroblast cells, and chicken bursa of Fabricius. The polyclonal system was more sensitive (P < 0.05) than the monoclonal system but both were specific as indicated by the negative results obtained with three non-IBDVs. The results revealed that the conventional systems used for titration of IBDVs (cell cultures and embryonating chicken eggs) were more sensitive than the polyclonal AC-ELISA.

  12. Estimating Mercury-Binding Ligand Concentrations in Freshwater Wetland Porewaters Using the “Tin-Reducible-Mercury” Titration Method

    NASA Astrophysics Data System (ADS)

    Creswell, J. E.; Babiarz, C.; Shafer, M. M.; Armstrong, D. E.

    2009-12-01

    Wetland environments are recognized as active regions of mercury methylation and may represent the primary source of methylmercury to many aquatic systems. Thus understanding the methylation process in these systems is vital to efforts at prediction of methylmercury accumulation at higher trophic levels. Strong mercury-binding ligands in porewaters can limit methylation in wetlands because large ligands and charged Hg-complexes are not bioavailable to methylating bacteria. Following the method developed by Lamborg and colleagues (2003), we used tin(II) chloride, a weak reductant, to reduce and measure labile Hg in porewater samples (i.e. the fraction not bound to strong ligands). We further titrated samples with Hg(II) to determine the Hg concentration at which the naturally occurring ligands were saturated - thus providing a measure of mercury-binding ligand concentrations. To our knowledge, this is the first extensive use of this technique on porewater samples. In an effort to differentiate between inorganic- and organic-complexed Hg, we modeled the inorganic speciation of Hg using MINEQL+, an aqueous speciation modeling program. Measurements of total mercury, methylmercury, sulfide, sulfate, chloride, carbonate, pH, dissolved organic carbon, phosphate, bromide, and major ions in sample porewaters were used as inputs to the model. Using the inorganic speciation calculated by the model, and the ligand concentrations measured in our laboratory experiment, we calculated conditional stability constants for mercury-ligand binding in this system. Although modeling predicts that mercury speciation will be dominated by sulfide at the concentrations present in this system, mercury methylation rate measurements show a stronger correlation with dissolved organic carbon concentrations than with sulfide. This correlation suggests that dissolved organic carbon plays an important role in mercury speciation, even in the presence of sulfide. Porewaters were extracted from

  13. Synthesis, potentiometric titration, electrochemical investigation and biological properties of trans-[RuCl2(dinic)4] (dinic = 3,5-pyridinecarboxylic acid).

    PubMed

    Seifriz, I; Konzen, M; Paula, M M; Gonçalves, N S; Spoganickz, B; Creczynski-Pasa, T B; Bonetti, V R; Beirith, A; Calixto, J B; Franco, C V

    1999-09-30

    This work discusses both the synthesis of trans-[RuCl2(dinic)4], dinic = 3,5-pyridinecarboxylic acid, and its main characteristics including potentiometric titration, spectroscopic and electrochemical properties, and some biological properties. The complex was synthesized using ruthenium blue solution as the precursor in a synthetic route. The complex was characterized using electronic spectroscopy, vibrational FT-IR spectroscopy, and Raman spectroscopy, as well as 1H and 13C NMR. The results indicated that the complex exhibits a trans-geometry. Cyclic voltammetry carried out in water:acetone 1:1 solution revealed a quasi-reversible process centered on the Ru(II) atom, as well as a dependence of the redox potential, E1/2, on pH. An analysis of the electronic spectra revealed that the MLCT (metal ligand charge transfer) band underwent a hypsochromic shift as the pH increased. Spectroelectrochemical analysis indicated that the visible region band progressively faded out upon oxidation. The equilibrium constants for the eight protons of the complex were determined by potentiometric titration. The complex neither inhibits the activity of nitrogen monoxide synthase nor acts as a scavenger for nitrogen monoxide. Nevertheless, the complex shows antinociceptive properties and acts as a scavenger for hydroxyl radicals.

  14. Reference method for total water in lint cotton by automated oven drying combined with volumetric Karl Fischer titration

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In a preliminary study to measure total water in lint cotton we demonstrated that volumetric Karl Fischer Titration of moisture transported by a carrier gas from an attached small oven is more accurate than standard oven drying in air. The objective of the present study was to assess the measuremen...

  15. Determination of sulphite in wine by coulometric titration.

    PubMed

    Lowinsohn, D; Bertotti, M

    2001-09-01

    A method for the determination of both free and bound S(IV) in white wine samples by coulometric titration with electrogenerated iodine is described, the S(IV) speciation being performed by extracting SO2 from samples acidified wiith hydrochloric acid. Titrations of samples treated with NaOH led to the total S(IV) concentration, the results being in agreement with the ones obtained by the distillation procedure as well as by using the classical Monier-Williams method. An average recovery of 97% was obtained for samples spiked with S(IV) in the range 0.4-1.2 mM, which demonstrated the validity of the proposed procedure. The repeatability was 1.3% and the limit of detection was estimated as 0.01 mM. Results from direct titrations of wine added to the electrochemical cell are reported, which give indications of the stability of adducts towards the reaction with iodine.

  16. Digital movie-based on automatic titrations.

    PubMed

    Lima, Ricardo Alexandre C; Almeida, Luciano F; Lyra, Wellington S; Siqueira, Lucas A; Gaião, Edvaldo N; Paiva Junior, Sérgio S L; Lima, Rafaela L F C

    2016-01-15

    This study proposes the use of digital movies (DMs) in a flow-batch analyzer (FBA) to perform automatic, fast and accurate titrations. The term used for this process is "Digital movie-based on automatic titrations" (DMB-AT). A webcam records the DM during the addition of the titrant to the mixing chamber (MC). While the DM is recorded, it is decompiled into frames ordered sequentially at a constant rate of 26 frames per second (FPS). The first frame is used as a reference to define the region of interest (ROI) of 28×13pixels and the R, G and B values, which are used to calculate the Hue (H) values for each frame. The Pearson's correlation coefficient (r) is calculated between the H values of the initial frame and each subsequent frame. The titration curves are plotted in real time using the r values and the opening time of the titrant valve. The end point is estimated by the second derivative method. A software written in C language manages all analytical steps and data treatment in real time. The feasibility of the method was attested by application in acid/base test samples and edible oils. Results were compared with classical titration and did not present statistically significant differences when the paired t-test at the 95% confidence level was applied. The proposed method is able to process about 117-128 samples per hour for the test and edible oil samples, respectively, and its precision was confirmed by overall relative standard deviation (RSD) values, always less than 1.0%.

  17. Digital movie-based on automatic titrations.

    PubMed

    Lima, Ricardo Alexandre C; Almeida, Luciano F; Lyra, Wellington S; Siqueira, Lucas A; Gaião, Edvaldo N; Paiva Junior, Sérgio S L; Lima, Rafaela L F C

    2016-01-15

    This study proposes the use of digital movies (DMs) in a flow-batch analyzer (FBA) to perform automatic, fast and accurate titrations. The term used for this process is "Digital movie-based on automatic titrations" (DMB-AT). A webcam records the DM during the addition of the titrant to the mixing chamber (MC). While the DM is recorded, it is decompiled into frames ordered sequentially at a constant rate of 26 frames per second (FPS). The first frame is used as a reference to define the region of interest (ROI) of 28×13pixels and the R, G and B values, which are used to calculate the Hue (H) values for each frame. The Pearson's correlation coefficient (r) is calculated between the H values of the initial frame and each subsequent frame. The titration curves are plotted in real time using the r values and the opening time of the titrant valve. The end point is estimated by the second derivative method. A software written in C language manages all analytical steps and data treatment in real time. The feasibility of the method was attested by application in acid/base test samples and edible oils. Results were compared with classical titration and did not present statistically significant differences when the paired t-test at the 95% confidence level was applied. The proposed method is able to process about 117-128 samples per hour for the test and edible oil samples, respectively, and its precision was confirmed by overall relative standard deviation (RSD) values, always less than 1.0%. PMID:26592600

  18. A high-throughput microtiter plate based method for the determination of peracetic acid and hydrogen peroxide.

    PubMed

    Putt, Karson S; Pugh, Randall B

    2013-01-01

    Peracetic acid is gaining usage in numerous industries who have found a myriad of uses for its antimicrobial activity. However, rapid high throughput quantitation methods for peracetic acid and hydrogen peroxide are lacking. Herein, we describe the development of a high-throughput microtiter plate based assay based upon the well known and trusted titration chemical reactions. The adaptation of these titration chemistries to rapid plate based absorbance methods for the sequential determination of hydrogen peroxide specifically and the total amount of peroxides present in solution are described. The results of these methods were compared to those of a standard titration and found to be in good agreement. Additionally, the utility of the developed method is demonstrated through the generation of degradation curves of both peracetic acid and hydrogen peroxide in a mixed solution. PMID:24260173

  19. A high-throughput microtiter plate based method for the determination of peracetic acid and hydrogen peroxide.

    PubMed

    Putt, Karson S; Pugh, Randall B

    2013-01-01

    Peracetic acid is gaining usage in numerous industries who have found a myriad of uses for its antimicrobial activity. However, rapid high throughput quantitation methods for peracetic acid and hydrogen peroxide are lacking. Herein, we describe the development of a high-throughput microtiter plate based assay based upon the well known and trusted titration chemical reactions. The adaptation of these titration chemistries to rapid plate based absorbance methods for the sequential determination of hydrogen peroxide specifically and the total amount of peroxides present in solution are described. The results of these methods were compared to those of a standard titration and found to be in good agreement. Additionally, the utility of the developed method is demonstrated through the generation of degradation curves of both peracetic acid and hydrogen peroxide in a mixed solution.

  20. Systemic errors in quantitative polymerase chain reaction titration of self-complementary adeno-associated viral vectors and improved alternative methods.

    PubMed

    Fagone, Paolo; Wright, J Fraser; Nathwani, Amit C; Nienhuis, Arthur W; Davidoff, Andrew M; Gray, John T

    2012-02-01

    Self-complementary AAV (scAAV) vector genomes contain a covalently closed hairpin derived from a mutated inverted terminal repeat that connects the two monomer single-stranded genomes into a head-to-head or tail-to-tail dimer. We found that during quantitative PCR (qPCR) this structure inhibits the amplification of proximal amplicons and causes the systemic underreporting of copy number by as much as 10-fold. We show that cleavage of scAAV vector genomes with restriction endonuclease to liberate amplicons from the covalently closed terminal hairpin restores quantitative amplification, and we implement this procedure in a simple, modified qPCR titration method for scAAV vectors. In addition, we developed and present an AAV genome titration procedure based on gel electrophoresis that requires minimal sample processing and has low interassay variability, and as such is well suited for the rigorous quality control demands of clinical vector production facilities.

  1. Determination of carbonate carbon in geological materials by coulometric titration

    USGS Publications Warehouse

    Engleman, E.E.; Jackson, L.L.; Norton, D.R.

    1985-01-01

    A coulometric titration is used for the determination of carbonate carbon in geological materials. Carbon dioxide is evolved from the sample by the addition of 2 M perchloric acid, with heating, and is determined by automated coulometric titration. The coulometric titration showed improved speed and precision with comparable accuracy to gravimetric and gasometric techniques. ?? 1985.

  2. Identification of Weak Acids and Bases by Titration with Primary Standards. A Modern Version of an Old Analytical Chemistry Experiment.

    ERIC Educational Resources Information Center

    Thompson, Robert Q.

    1988-01-01

    Describes a laboratory exercise in which acid dissociation constants and molecular weights are extracted from sample data and the sample is identified. Emphasizes accurate volumetric work while bringing to practice the concepts of acid-base equilibria, activity coefficients, and thermodynamic constants. (CW)

  3. On-line dilution and determination of the amount of concentrated hydrochloric acid in the final products from a hydrochloric acid production plant using a sequential injection titration system.

    PubMed

    van Staden, J Koos F; Mashamba, Mulalo G; Stefan, Raluca I

    2002-12-01

    An on-line sequential injection titration system for the determination of the concentration of concentrated hydrochloric acid as final product from a hydrochloric acid production plant is described. The system involves on-line dilution of the concentrated hydrochloric acid solution to an acceptable range for direct measurement by merging the sample stream with a de-ionized water diluent stream, followed by mixing in a dilution coil, before aspiration into the sequential injection system. Concentrated standard solutions were treated in exactly the same way as the samples. The system was evaluated for reproducibility, linearity, accuracy, and sample throughput. A linear relationship between peak width and logarithm of acid concentration was found in the range 5.934-8.995 mol l(-1) and a concentration of 0.005 mol l(-1) NaOH solution was used as titrant. Samples from the production plant showed excellent agreement when compared with the manual and automated batchwise titrations. The relative standard deviation was found to be less than 0.4% with a sample frequency of 30 samples per hour.

  4. Effect of retinoic acid on HPV titration and colposcopic changes in Korean patients with dysplasia of the uterine cervix.

    PubMed

    Ahn, W S; Lee, J M; Namkoong, S E; Lee, H Y; Kim, S J

    1997-01-01

    Retinoids, a family of molecules capable of profound impact on many biological functions, have antiproliferative, differentiative, and immunomodulatory properties. The present study assessed the effect of 13-cis-retinoic acid (13-CRA) treatment in 13 chronic cervicitis and 52 cervical intraepithelial neoplasia patients. We examined low- and high-risk human papilloma virus titer (using the hybrid capture method) and made a colposcopic and cervicographic examination before and after treatment with 13-CRA at 1 mg/kg for 4 to 12 weeks. Patients were between 27 and 64 years, the average age being 36.6 years. Histology revealed chronic cervicitis in 13 cases, mild dysplasia in 18 cases, moderate dysplasia in 18 cases, and severe dysplasia in 16 cases, totaling 65 cases. The expression rate of high-risk human papilloma virus (HPV 16, 18) was 9 of 13 cases (69%) in chronic cervicitis, 7 of 18 cases (38%) in mild dysplasia, 9 of 18 cases (50%) in moderate dysplasia, and 12 of 16 cases (75%) in severe dysplasia, with the overall expression rate being 37 of 65 cases (57%). Following 13-CRA treatment, decreases in high-risk titer were observed in 6 of 9 cases (66%) of chronic cervicitis, 4 of 11 cases (36%) of mild dysplasia, 7 of 9 cases (77%) of moderate dysplasia, and 8 of 12 cases (75%) of severe dysplasia. Overall, HPV titer decreased in 25 of 41 cases (61%). Minimal changes were found in colposcopic and cervicographic observations during the study. In summary, high-risk HPV titer decreased after treatment with 13-CRA in the majority of patients with cervical intraepithelial neoplasia. This study supports the potential of retinoids to interrupt multi-step carcinogenesis, possibly by down-regulation of gene products (E6,E7) produced by HPV infection.

  5. Analysis of the purity of cetrimide by titrations.

    PubMed

    Rasmussen, Claus D; Nielsen, Hans B; Andersen, Jens E T

    2006-01-01

    The purity of cetrimide, trimethyl tetradecyl ammonium bromide (TTAB), which is an important preservative of many cosmetic and pharmaceutical products, was determined by three independent methods of titration. Traditionally, cetrimide was analysed by an assay method of the European Pharmacopoeia, which showed consistently a low purity of cetrimide with associated large standard deviations, however. A systematic 3% bias of the European Pharmacopoeia assay method was identified by comparing the result with results of two alternative methods of titration that exhibited high precision and high accuracy. Titration by perchloric acid showed a 99.69% +/- 0.05% purity of cetrimide, and titration by silver nitrate showed a 99.85% +/- 0.05% purity, while the traditional assay method predicted a purity of only 97.1% +/- 0.4%. It was found that the discrepancy could be identified as differences in selectivity during the extraction step of the European Pharmacopoeia assay method. The distribution coefficients between chloroform and water of cetrimide and the corresponding iodide species (TTAI) were thus determined as 2150 +/- 50 M(-1) and 68000 +/- 4000 M(-1), respectively.

  6. Hydrogen bonding and molecular association in 2-(quinuclidinium)-butyric acid bromide hydrate studied by X-ray diffraction, DFT calculations, FTIR and NMR spectroscopy, and potentiometric titration

    NASA Astrophysics Data System (ADS)

    Dega-Szafran, Z.; Katrusiak, A.; Szafran, M.; Barczyński, P.

    2010-06-01

    The structure of 2-(quinuclidinium)-butyric acid bromide hydrate (QNBu·H 2O·HBr, 3) has been determined by X-ray diffraction, DFT calculations and characterized by FTIR and NMR spectroscopy. Crystals of 3 are monoclinic, space group P2 1. The water molecule interacts with the carboxylic group of 2-(quinuclidinium)-butyric acid and with the bromide anion by the COOH⋯OH 2 and HOH⋯Br hydrogen bonds of 2.575(3) and 3.293(2) Å, respectively. The structures of monomer ( 4) and dimeric cation ( 5) of the title complex have been optimized by the B3LYP/6-31G(d,p) approach, yielding conformations consistent with this in the crystal. The solid-state FTIR spectra of 3 and its deuterated analogue have been measured and compared with the theoretical spectrum of 4. The assignments of the observed and predicted bands have been proposed. The molecule of 3 has a chiral center at the C(9) atom, which is responsible for the non-magnetically equivalence of the α-ring and C(11)H 2 methylene protons in 1H NMR spectrum. The values of p Ka of quinuclidinium-acetate (quinuclidine betaine), 2-(quinuclidinium)-propionate and 2-(quinuclidinium)-butyrate have been determined by the potentiometric titration of their hydrohalides.

  7. Potentiometric titrations using pencil and graphite sensors

    SciTech Connect

    Selig, W.S.

    1984-01-01

    The cost of various commercial indicating electrodes ranges from about $40 for pH electrodes to as much as $355 for a potassium ion-selective electrode. This cost can be reduced to less than $1.50, and in some cases to mere pennies by making sensors from graphite rods and pencils for use in potentiometric titrations. The same sensor can be used for many types of these titrations (acid/base, compleximetric, precipitation, and redox). 8 references, 2 tables.

  8. Modeling complexometric titrations of natural water samples.

    PubMed

    Hudson, Robert J M; Rue, Eden L; Bruland, Kenneth W

    2003-04-15

    Complexometric titrations are the primary source of metal speciation data for aquatic systems, yet their interpretation in waters containing humic and fulvic acids remains problematic. In particular, the accuracy of inferred ambient free metal ion concentrations and parameters quantifying metal complexation by natural ligands has been challenged because of the difficulties inherent in calibrating common analytical methods and in modeling the diverse array of ligands present. This work tests and applies a new method of modeling titration data that combines calibration of analytical sensitivity (S) and estimation of concentrations and stability constants for discrete natural ligand classes ([Li]T and Ki) into a single step using nonlinear regression and a new analytical solution to the one-metal/two-ligand equilibrium problem. When applied to jointly model data from multiple titrations conducted at different analytical windows, it yields accurate estimates of S, [Li]T, Ki, and [Cu2+] plus Monte Carlo-based estimates of the uncertainty in [Cu2+]. Jointly modeling titration data at low-and high-analytical windows leads to an efficient adaptation of the recently proposed "overload" approach to calibrating ACSV/CLE measurements. Application of the method to published data sets yields model results with greater accuracy and precision than originally obtained. The discrete ligand-class model is also re-parametrized, using humic and fulvic acids, L1 class (K1 = 10(13) M(-1)), and strong ligands (L(S)) with K(S) > K1 as "natural components". This approach suggests that Cu complexation in NW Mediterranean Sea water can be well represented as 0.8 +/- 0.3/0.2 mg humic equiv/L, 13 +/- 1 nM L1, and 2.5 +/- 0.1 nM L(S) with [CU]T = 3 nM. In coastal seawater from Narragansett Bay, RI, Cu speciation can be modeled as 0.6 +/- 0.1 mg humic equiv/L and 22 +/- 1 nM L1 or approximately 12 nM L1 and approximately 9 nM L(S), with [CU]T = 13 nM. In both waters, the large excess

  9. Copper redox transformation and complexation by reduced and oxidized soil humic Acid. 2. Potentiometric titrations and dialysis cell experiments.

    PubMed

    Maurer, Felix; Christl, Iso; Fulda, Beate; Voegelin, Andreas; Kretzschmar, Ruben

    2013-10-01

    Cation binding and electron transfer reactions of humic substances determine copper speciation in redox-dynamic systems, but quantitative studies for Cu+ binding to humic substances are lacking. We investigated reduction of Cu2+ and binding of Cu+ at pH 7.0 in a dialysis cell experiment using reduced and reoxidized soil humic acid (HA) as reductant and sorbent at copper loadings of 9.5-600 mmol kg(-1). The data were used to quantitatively explain the interaction between cation binding and electron transfer processes that determine copper speciation in the presence of HA under anoxic and oxic conditions. Addition of Cu2+ to reduced HA resulted in almost complete reduction to Cu(I) within 1 h. Reduction was also significant under oxic conditions. The slow decrease of the Cu(I) fraction was attributed to formation of Cu(0) based on thermodynamic consideration. Cu+ binding to HA was found to be strong compared to other chalcophile cations like Ag+ or Cd2+. Our results indicate that Cu+ and Cu2+ isotherms exhibit a redox potential-dependent intersection point. According to the differences in Cu+ and Cu2+ binding, the presence of HA was found to extend the stability field of Cu(II) to moderately reducing conditions and to reduce the stability field of Cu(0) due to the formation of Cu(I) complexes.

  10. Comparative assessment of the methods for exchangeable acidity measuring

    NASA Astrophysics Data System (ADS)

    Vanchikova, E. V.; Shamrikova, E. V.; Bespyatykh, N. V.; Zaboeva, G. A.; Bobrova, Yu. I.; Kyz"yurova, E. V.; Grishchenko, N. V.

    2016-05-01

    A comparative assessment of the results of measuring the exchangeable acidity and its components by different methods was performed for the main mineral genetic horizons of texturally-differentiated gleyed and nongleyed soddy-podzolic and gley-podzolic soils of the Komi Republic. It was shown that the contents of all the components of exchangeable soil acidity determined by the Russian method (with potassium chloride solution as extractant, c(KCl) = 1 mol/dm3) were significantly higher than those obtained by the international method (with barium chloride solution as extractant, c(BaCl2) = 0.1 mol/dm3). The error of the estimate of the concentration of H+ ions extracted with barium chloride solution equaled 100%, and this allowed only qualitative description of this component of the soil acidity. In the case of the extraction with potassium chloride, the error of measurements was 50%. It was also shown that the use of potentiometric titration suggested by the Russian method overestimates the results of soil acidity measurement caused by the exchangeable metal ions (Al(III), Fe(III), and Mn(II)) in comparison with the atomic emission method.

  11. Micro coulometric titration in a liquid drop.

    PubMed

    Kanyanee, Tinakorn; Fuekhad, Pongwasin; Grudpan, Kate

    2013-10-15

    Miniaturized coulometric titration in a liquid drop has been investigated. Assays of ascorbic acid and thiosulfate with iodine titration were chosen as models. Constant volumes of falling liquid drops containing sample or reagent are manipulated via gravimetrical force to move along a slope hydrophobic path and directed to stop or to move out from an electrode. Such manipulation is useful for delivery of sample and reagents, in a way of flow without tubing. Electrochemical generation of titrant, in this case, iodine, is started at the electrode and micro coulometric titration can be performed in a drop by applying constant current. Timing in the titration can be made via naked eye with a stopwatch or via recording with a webcam camera connecting to a computer to detect the change due to the blue color complex of the excess iodine and starch.

  12. Micro coulometric titration in a liquid drop.

    PubMed

    Kanyanee, Tinakorn; Fuekhad, Pongwasin; Grudpan, Kate

    2013-10-15

    Miniaturized coulometric titration in a liquid drop has been investigated. Assays of ascorbic acid and thiosulfate with iodine titration were chosen as models. Constant volumes of falling liquid drops containing sample or reagent are manipulated via gravimetrical force to move along a slope hydrophobic path and directed to stop or to move out from an electrode. Such manipulation is useful for delivery of sample and reagents, in a way of flow without tubing. Electrochemical generation of titrant, in this case, iodine, is started at the electrode and micro coulometric titration can be performed in a drop by applying constant current. Timing in the titration can be made via naked eye with a stopwatch or via recording with a webcam camera connecting to a computer to detect the change due to the blue color complex of the excess iodine and starch. PMID:24054589

  13. Students' integration of multiple representations in a titration experiment

    NASA Astrophysics Data System (ADS)

    Kunze, Nicole M.

    A complete understanding of a chemical concept is dependent upon a student's ability to understand the microscopic or particulate nature of the phenomenon and integrate the microscopic, symbolic, and macroscopic representations of the phenomenon. Acid-base chemistry is a general chemistry topic requiring students to understand the topics of chemical reactions, solutions, and equilibrium presented earlier in the course. In this study, twenty-five student volunteers from a second semester general chemistry course completed two interviews. The first interview was completed prior to any classroom instruction on acids and bases. The second interview took place after classroom instruction, a prelab activity consisting of a titration calculation worksheet, a titration computer simulation, or a microscopic level animation of a titration, and two microcomputer-based laboratory (MBL) titration experiments. During the interviews, participants were asked to define and describe acid-base concepts and in the second interview they also drew the microscopic representations of four stages in an acid-base titration. An analysis of the data showed that participants had integrated the three representations of an acid-base titration to varying degrees. While some participants showed complete understanding of acids, bases, titrations, and solution chemistry, other participants showed several alternative conceptions concerning strong acid and base dissociation, the formation of titration products, and the dissociation of soluble salts. Before instruction, participants' definitions of acid, base, and pH were brief and consisted of descriptive terms. After instruction, the definitions were more scientific and reflected the definitions presented during classroom instruction.

  14. A PC-based titrator for flow gradient titrations

    PubMed Central

    Fuhrmann, B.

    1993-01-01

    This paper describes a PC (personal computer) based titrator which was developed for gradient flow titrations. Concentration gradients were generated electrolytically or volumetrically in small tubes. Complete titration curves can be recorded on-line and evaluated automatically. The titrator can be used with all liquid flow detectors with low axial dispersion. The titrator was evaluated for the titration of thiosulphate with electrogenerated triiodide and for the titration of ammonia with electrogenerated hypobromite after continuous gas dialytic separation of ammonia from the sample solution. PMID:18924977

  15. Gasometric titration for dimethylaluminum chloride analysis.

    PubMed

    Wang, Lin; Maligres, Peter; Eckenroad, Kyle; Simmons, Bryon

    2016-06-01

    A gasometric titration method was developed to quantitate active alkylaluminum content in dimethylaluminum chloride solution to perform the stoichiometry calculation for the reaction charge. The procedure was reproducible with good precision, and the results showed good correlation with ICP-MS method. The gasometric titration is a simple, inexpensive alternative to analysis via ICP-MS which provides more selective analysis of methylaluminum species without the need for inertion.

  16. Well acidizing compositions and methods

    SciTech Connect

    Swanson, B. L.

    1980-12-23

    Gelled acidic compositions suitable for matrix acidizing or fracture acidizing of subterranean formations are provided comprising water, a water-dispersible polymeric viscosifier such as a polymer of acrylamide, an acid, and a polyphenolic material such as lignite.

  17. Nile blue and brilliant cresyl blue as redox indicators in iron(II) titrations.

    PubMed

    Sriramam, K

    1975-01-01

    Nile Blue and Brilliant Cresyl Blue, two compounds related to diaminophenoxazine, have been studied as indicators in titrations of iron(II) with cerium(IV)(in hydrochloric, sulphuric and perchloric acid media), dichromate, vanadate and permanganate. They are particularly suited for titrations in a fairly concentrated sulphuric acid medium and for titrations with dilute solutions. A probable indicator mechanism is suggested.

  18. Determination of sulfur compounds in hydrotreated transformer base oil by potentiometric titration.

    PubMed

    Chao, Qiu; Sheng, Han; Cheng, Xingguo; Ren, Tianhui

    2005-06-01

    A method was developed to analyze the distribution of sulfur compounds in model sulfur compounds by potentiometric titration, and applied to analyze hydrotreated transformer base oil. Model thioethers were oxidized to corresponding sulfoxides by tetrabutylammonium periodate and sodium metaperiodate, respectively, and the sulfoxides were titrated by perchloric acid titrant in acetic anhydride. The contents of aliphatic thioethers and total thioethers were then determined from that of sulfoxides in solution. The method was applied to determine the organic sulfur compounds in hydrotreated transformer base oil.

  19. Comparison between dot-immunoblotting assay and clinical sign determination method for quantifying avian infectious bronchitis virus vaccine by titration in embryonated eggs.

    PubMed

    Yuk, Seong-su; Kwon, Jung-Hoon; Noh, Jin-Yong; Hong, Woo-tack; Gwon, Gyeong-Bin; Jeong, Jei-Hyun; Jeong, Sol; Youn, Ha-Na; Heo, Yong-Hwan; Lee, Joong-Bok; Park, Seung-Yong; Choi, In-Soo; Song, Chang-Seon

    2016-04-01

    A sensitive and specific method for measuring the vaccine titer of infectious bronchitis virus (IBV) is important to commercial manufacturers for improving vaccine quality. Typically, IBV is titrated in embryonated chicken eggs, and the infectivity of the virus dilutions is determined by assessing clinical signs in the embryos as evidence of viral propagation. In this study, we used a dot-immunoblotting assay (DIA) to measure the titers of IBV vaccines that originated from different pathogenic strains or attenuation methods in embryonated eggs, and we compared this assay to the currently used method, clinical sign evaluation. To compare the two methods, we used real-time reverse transcription-PCR, which had the lowest limit of detection for propagated IBV. As a clinical sign of infection, dwarfism of the embryo was quantified using the embryo: egg (EE) index. The DIA showed 9.41% higher sensitivity and 15.5% higher specificity than the clinical sign determination method. The DIA was particularly useful for measuring the titer of IBV vaccine that did not cause apparent stunting but propagated in embryonated chicken eggs such as a heat-adapted vaccine strain. The results of this study indicate that the DIA is a rapid, sensitive, reliable method for determining IBV vaccine titer in embryonated eggs at a relatively low cost.

  20. The Titration Project.

    ERIC Educational Resources Information Center

    Kilner, Cary

    1988-01-01

    Discusses the development of concentration and organizational skills, patience, self-discipline, attention to detail, and appreciation for error analysis through an expanded titration project. Describes the various activities in the extended project and the materials and instructional support needed. Stresses the advantage to students in their…

  1. Titration Calculations with Computer Algebra Software

    ERIC Educational Resources Information Center

    Lachance, Russ; Biaglow, Andrew

    2012-01-01

    This article examines the symbolic algebraic solution of the titration equations for a diprotic acid, as obtained using "Mathematica," "Maple," and "Mathcad." The equilibrium and conservation equations are solved symbolically by the programs to eliminate the approximations that normally would be performed by the student. Of the three programs,…

  2. Problems in the interpretation of nonaqueous titrations.

    PubMed

    Buvári-Barcza, A; Barcza, L

    2005-04-01

    Most phenomena (like promoting and leveling as well as differentiating effects) of common nonaqueous titrations can be explained by the Brönsted's concept combined with the hydrogen bond theory but some open problems are shown in connection with the use of formic acid and/or acetic anhydride.

  3. Nucleic acid arrays and methods of synthesis

    DOEpatents

    Sabanayagam, Chandran R.; Sano, Takeshi; Misasi, John; Hatch, Anson; Cantor, Charles

    2001-01-01

    The present invention generally relates to high density nucleic acid arrays and methods of synthesizing nucleic acid sequences on a solid surface. Specifically, the present invention contemplates the use of stabilized nucleic acid primer sequences immobilized on solid surfaces, and circular nucleic acid sequence templates combined with the use of isothermal rolling circle amplification to thereby increase nucleic acid sequence concentrations in a sample or on an array of nucleic acid sequences.

  4. Standard test method for water in lint cotton by oven evaporation combined with volumetric Karl Fischer Titration

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The referenced test method for total water content and water regain in lint cotton was developed by USDA scientists in New Orleans at the request of the cotton industry. The method covers the determination of the total water (free and bound) in raw and lint cotton at moisture equilibrium from con...

  5. A microtiter test for detecting and titrating noncytopathogenic bovine viral diarrhea virus.

    PubMed

    Maisonnave, J; Rossi, C R

    1982-01-01

    Bovine cells free of noncytopathogenic bovine viral diarrhea virus (NC-BVDV) treated with polyriboinosinic acid : polyribocytidylic acid (poly I:C) were protected against challenge with vesicular stomatitis virus (VSV), whereas NC-BVDV-infected cells treated with poly I:C were not protected against VSV. An assay based on the ability of NC-BVDV to inhibit poly I:C protection of cells against VSV was developed and is herein referred to as PINBA (poly I:C for NC-BVDV assay). Noncytopathogenic BVDV was titrated as cytopathogenic strains except that several days after infection with NC-BVDV, the cultures were treated with poly I:C and VSV. Titration endpoints were reached 24 hours later. PINBA was standardized for amount of VSV, time of addition of poly I:C, and time NC-BVDV had to be present to obtain stable titration endpoints. PINBA also was useful for titrating virus neutralizing antibodies. Compared with the fluorescent antibody test, PINBA was less subjective for detection of NC-BVDV. Compared with the interference test in which NC-BVDV infected cultures are challenged with a cytopathogenic strain of BVDV, PINBA was more reliable. The technique described herein is a simple and practical microtiter method for titrating NC-BVDV and virus neutralizing antibodies and for the presumptive detection of NC-BVDV.

  6. Micro method for performing titration and neutralization test of hog cholera virus using established porcine kidney cell strain.

    PubMed

    Komaniwa, H; Fukusho, A; Shimizu, Y

    1981-01-01

    Hog cholera (HC) virus and antibody against it were estimated by the END method with microplates and CPK porcine kidney cell strain. To establish the technique of this method, studies were made on such basic conditions of the method as the type of strain of Newcastle disease virus (NDV), the time of challenge with this virus, and the concentration of serum in culture fluid. There was little difference in the infective titer of HC virus estimated between the END method performed by the established technique and the same method by the conventional technique with test tubes and swine testicle (ST) cells. Besides, there was a high correlation between the neutralizing antibody titer measured by the one technique and that measured by the other. The coefficient of correlation was r = +0.948 in this case. From the experimental results mentioned above it was concluded that the END method by the micro-technique with CPK cells was simpler than and as reliable as the same method conducted by any conventional technique, and that it was a practicable one capable of testing many samples.

  7. Determination of metal speciation by reverse titrations.

    PubMed

    Nuester, Jochen; van den Berg, Constant M G

    2005-01-01

    A new method is proposed to determine metal speciation by varying the concentration of a competing ligand at a constant metal concentration, with detection by cathodic stripping voltammetry. The free metal ion concentration is gradually lowered from its natural level while the method probes progressively deeper into the already complexed metal fraction: it is therefore a reverse titration rather than the forward titration which is used for conventional complexing ligand titrations. The sensitivity is greatest at the lowest free metal ion concentration, where it matters most, and the method can be carried out in a single sample aliquot in the voltammetric cell. The method is applied here to copper speciation, but in principle, it can also be used for other metals. Modeling shows that this method has good sensitivity at ligand concentrations near the metal concentration (lower as well as greater). Comparative measurements of copper speciation using reverse and forward titrations of representative water samples of oceanic and coastal origin show good agreement. The data showed that these samples did not contain low levels of strong ligands in addition to the ligands detected by the forward titrations.

  8. The molecular mechanism of the neutral-to-base transition of human serum albumin. Acid/base titration and proton nuclear magnetic resonance studies on a large peptic and a large tryptic fragment of albumin.

    PubMed

    Bos, O J; Labro, J F; Fischer, M J; Wilting, J; Janssen, L H

    1989-01-15

    In order to obtain a better understanding of the neutral-to-base (N-B) transition of human serum albumin, we performed acid/base titration experiments and 500-MHz 1H NMR experiments on albumin and on a large peptic (residues 1-387) and large tryptic (residues 198-585) fragment of albumin. The acid/base titration experiments revealed that Ca2+ ions induce a downward pK shift of several histidine residues of the peptic (P46) fragment and of albumin. By contrast, Ca2+ has very little influence on the pK of histidine residues of the tryptic (T45) fragment. In albumin, the pH-dependent His C-2 proton resonances, observed with 1H NMR experiments, have been allotted the numbers 1-17. It proved possible to locate these resonances in the P46 and the T45 fragments. A correspondence was found between the number of histidines detected by the acid/base titration and by the 1H NMR experiments. The results of the experiments lead us to conclude that in domain 1 at least the histidines corresponding to the His C-2 proton resonances 1-5 play a dominant role in the N-B transition. The Cu2+-binding histidine residue 3 (resonance 8) of the albumin molecule is not involved in the N-B transition. In addition, we were able to assign His C-2 proton resonance 9 to histidine 464 of the albumin molecule. The role of the N-B transition in the transport and cellular uptake mechanisms of endogenous and exogenous compounds is discussed.

  9. Differential Binding Models for Direct and Reverse Isothermal Titration Calorimetry.

    PubMed

    Herrera, Isaac; Winnik, Mitchell A

    2016-03-10

    Isothermal titration calorimetry (ITC) is a technique to measure the stoichiometry and thermodynamics from binding experiments. Identifying an appropriate mathematical model to evaluate titration curves of receptors with multiple sites is challenging, particularly when the stoichiometry or binding mechanism is not available. In a recent theoretical study, we presented a differential binding model (DBM) to study calorimetry titrations independently of the interaction among the binding sites (Herrera, I.; Winnik, M. A. J. Phys. Chem. B 2013, 117, 8659-8672). Here, we build upon our DBM and show its practical application to evaluate calorimetry titrations of receptors with multiple sites independently of the titration direction. Specifically, we present a set of ordinary differential equations (ODEs) with the general form d[S]/dV that can be integrated numerically to calculate the equilibrium concentrations of free and bound species S at every injection step and, subsequently, to evaluate the volume-normalized heat signal (δQ(V) = δq/dV) of direct and reverse calorimetry titrations. Additionally, we identify factors that influence the shape of the titration curve and can be used to optimize the initial concentrations of titrant and analyte. We demonstrate the flexibility of our updated DBM by applying these differentials and a global regression analysis to direct and reverse calorimetric titrations of gadolinium ions with multidentate ligands of increasing denticity, namely, diglycolic acid (DGA), citric acid (CIT), and nitrilotriacetic acid (NTA), and use statistical tests to validate the stoichiometries for the metal-ligand pairs studied.

  10. Biophysical studies on the interactions of a classic mitochondrial uncoupler with bovine serum albumin by spectroscopic, isothermal titration calorimetric and molecular modeling methods.

    PubMed

    Zhang, Yue; Li, Jia-Han; Ge, Yu-Shu; Liu, Xiao-Rong; Jiang, Feng-Lei; Liu, Yi

    2011-03-01

    The interaction between a classic uncoupler (2,4-dinitrophenol, DNP) and bovine serum albumin (BSA) was investigated by fluorescence spectroscopy under the physiological conditions. The fluorescence quenching constants were calculated by the Stern-Volmer equation, and based upon the temperature dependence of quenching constants, it was proved that DNP caused a static quenching of the intrinsic fluorescence of BSA. Owing to the static quenching mechanism, different associative binding constants at various temperatures were determined and thus the thermodynamic parameters, namely enthalpy (ΔH=-21.12 kJ mol(-1)) and entropy changes (ΔS=23.51 J mol(-1) K(-1)) could be calculated based on the binding constants. Moreover, the enthalpy and entropy changes are consistent with the "Enthalpy-Entropy Compensation" equation obtained from our previous work. The negative enthalpy and positive entropy indicated that the electrostatic interactions played a major role in DNP-BSA binding process. Site marker competitive displacement experiments were carried out by using fluorescence and isothermal titration calorimetry (ITC) methods. These results showed that DNP bound with high affinity to Sudlow's site I (subdomain IIA) of BSA. The distance (r=3.78 nm) between donor (BSA) and acceptor (DNP) was obtained according to the mechanism of fluorescence resonance energy transfer (FRET). Furthermore, the results of synchronous fluorescence and circular dichroism (CD) spectroscopic studies indicated that the microenvironment and the secondary conformation of BSA were altered. The above results were supported by theoretical molecular modeling methods.

  11. Comparison of neuraminidases of the same subtype but from different species using a new method of titration.

    PubMed

    Fiszon, B; Hannoun, C

    1990-01-01

    Neuraminidase is one of the two surface glycoproteins of influenza virions. In order to compare neuraminidases of the same subtype but isolated from different species (man, birds, pig), a new and simple method was adapted and optimized using peanut hemagglutinin. Results were very similar to those obtained with the classical method recommended by the WHO, using fetuin as a substrate. The technique was used to examine the relationship between animal and human neuraminidases belonging to serotypes N1 and N2. The results confirm the possible role of ducks as a reservoir for influenza viruses and the eventuality of interspecific exchanges.

  12. [Alkalimetric titrations of salts of organic bases in the Pharmacopoeia].

    PubMed

    Bezáková, Zelmíra; Stankovičová, Mária

    2013-12-01

    Modified methods - alkalimetry in ethanol 70% with a defined small volume of hydrochloric acid 0.01 mol/l added to the solution of the sample before the titration and alkalimetry in ethanol 70% or ethanol 96% alone with potentiometric end-point detection for the assay of halide salts of 11 organic N-bases has been investigated. The results were compared to those obtained by the method of the European Pharmacopoeia 7th Ed. (Ph. Eur. 7th Ed.). The Ph. Eur. 7th Ed. use for 8 investigated substances alkalimetry in alcohol 96 % with a defined small volume of hydrochloric acid 0.01 mol/l (5 ml) with potentiometric end-point detection: Cinchocaine hydrochloride, Codeine hydrochloride dihydrate, Ethylmorphine hydrochloride, Lidocaine hydrochloride, Papaverine hydrochloride, Pilocarpine hydrochloride, Quinine hydrochloride, Tetracaine hydrochloride. Our results revealed that the Ph. Eur. 7th Ed. method did not work for 5 drugs from this group: Cinchocaine hydrochloride, Ethylmorphine hydrochloride, Papaverine hydrochloride, Pilocarpine hydrochloride and Tetracaine hydrochloride. In the group of investigated substances we included also drugs with the character of weak organic bases for which Ph. Eur. 7th Ed. prescribed different methods for their assay: Thiamine hydrochloride and Pyridoxine hydrochloride - acidimetric titration in non-aqueous solvents with perchloric acid and Procaine hydrochloride - determination of primary aromatic amino-nitrogen (Ph. Eur. 7th Ed., chapter 2.5.8).

  13. Routine titration of foot-and-mouth disease virus suspensions by analytical ultracentrifugation 2nd communication: sedimentation equilibrium method.

    PubMed

    Strobbe, R; Lacroix, C; Charlier, G; Debecq, J

    1977-01-01

    A routine method for the determination of the virus concentration in FMD virus cultures and vaccines was developed. This method was based on sedimentation equilibrium in the analytical ultraviolet scanning ultracentrifuge. The virus suspension was first clarified. The virions were then sedimented in a preparative ultracentrifuge. The resuspended virions were diluted in a Cesium chloride solution and brought to equilibrium in the density gradient generated in the analytical ultracentrifuge. The optical density of the virus band was measured by the UV scanning system. A calculation procedure was developed to compute the density at the limits and at the maximum of the virus band. The virus concentration expressed as weight, was calculated for the original virus suspension.

  14. [Titration of Ebola and Marburg viruses by plaque formation under semi liquid agar].

    PubMed

    Ustinova, E N; Shestopalov, A M; Bakulina, L F; Chepurnov, A A

    2003-01-01

    The method of titration of Ebola and Marburg viruses using plaque formation under semifluid agar cover is considered. Advantages of this method over conventional method of titration of these viruses with the use of hard agar cover are discussed.

  15. Microculture virus titration--a simple colourimetric assay for influenza virus titration.

    PubMed

    Levi, R; Beeor-Tzahar, T; Arnon, R

    1995-03-01

    Influenza antigens can be detected by several well established methods. However, when it is important to determine the titre of infective virions, a bioassay should be employed. The standard and the most widely used tests for influenza infectivity are titration carried out in embryonated hen eggs, or the plaque assay employing tissue culture techniques. A simple colourimetric assay for influenza virus detection and titration is described. Samples of allantoic fluid or mice lung homogenates were used to infect MDCK cultures in microplate wells. After an incubation period, the tetrazolium (MTT) colourimetric assay was used to determine cell viability, and when compared to untreated culture control enabled the detection and titration of several influenza strains. When samples were assayed simultaneously in embryonated eggs and by the MCVT method, good correlation in determined titres was obtained. The availability of an additional method for influenza titration allows more flexibility in the choice of titration method according to the specific needs of the study. Furthermore, this method lends itself to full automatization. Similar procedures should also be applicable to titration of other cytopathic viruses.

  16. 40 CFR Appendix A-4 to Part 60 - Test Methods 6 through 10B

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... Maintenance Requirements) of 40 CFR part 60, subpart A (General Provisions). Specific uses of these test... sulfur acid mist, are separated. The SO2 fraction is measured by the barium-thorin titration method. 3... normality determination 11.2.3 Replicate titrations Ensure precision of titration determinations....

  17. 40 CFR Appendix A-4 to Part 60 - Test Methods 6 through 10B

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... Maintenance Requirements) of 40 CFR part 60, subpart A (General Provisions). Specific uses of these test... sulfur acid mist, are separated. The SO2 fraction is measured by the barium-thorin titration method. 3... normality determination 11.2.3 Replicate titrations Ensure precision of titration determinations....

  18. 40 CFR Appendix A-4 to Part 60 - Test Methods 6 through 10B

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... Maintenance Requirements) of 40 CFR part 60, subpart A (General Provisions). Specific uses of these test... sulfur acid mist, are separated. The SO2 fraction is measured by the barium-thorin titration method. 3... normality determination. 11.2.3 Replicate titrations Ensure precision of titration determinations 11.3...

  19. 40 CFR Appendix A-4 to Part 60 - Test Methods 6 through 10B

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... Maintenance Requirements) of 40 CFR part 60, subpart A (General Provisions). Specific uses of these test... sulfur acid mist, are separated. The SO2 fraction is measured by the barium-thorin titration method. 3... normality determination 11.2.3 Replicate titrations Ensure precision of titration determinations....

  20. Probing the binding mechanisms of α-tocopherol to trypsin and pepsin using isothermal titration calorimetry, spectroscopic, and molecular modeling methods.

    PubMed

    Li, Xiangrong; Ni, Tianjun

    2016-06-01

    α-Tocopherol is a required nutrient for a variety of biological functions. In this study, the binding of α-tocopherol to trypsin and pepsin was investigated using isothermal titration calorimetry (ITC), steady-state and time-resolved fluorescence measurements, circular dichroism (CD) spectroscopy, and molecular modeling methods. Thermodynamic investigations reveal that α-tocopherol binds to trypsin/pepsin is synergistically driven by enthalpy and entropy. The fluorescence experimental results indicate that α-tocopherol can quench the fluorescence of trypsin/pepsin through a static quenching mechanism. The binding ability of α-tocopherol with trypsin/pepsin is in the intermediate range, and one molecule of α-tocopherol combines with one molecule of trypsin/pepsin. As shown by circular dichroism (CD) spectroscopy, α-tocopherol may induce conformational changes of trypsin/pepsin. Molecular modeling displays the specific binding site and gives information about binding forces and α-tocopherol-tryptophan (Trp)/tyrosine (Tyr) distances. In addition, the inhibition rate of α-tocopherol on trypsin and pepsin was studied. The study provides a basic data set for clarifying the binding mechanisms of α-tocopherol with trypsin and pepsin and is helpful for understanding its biological activity in vivo.

  1. Agreement between the cell culture titrations of canine minute virus determined by two susceptibility-testing methods.

    PubMed

    Pratelli, A; Colao, V

    2013-09-01

    The correct diagnosis of canine minute virus is critical in dog breeding. In this study, the Bland Altman test was used to compare the performance of two susceptibility-testing methods, namely polymerase chain reaction (PCR) and indirect immunofluorescence assay (IFA). The agreement between IFA and PCR in monocytes revealed a mean difference of -1752.16 with 95% confidence and an interval ranging from -3229.80 to -274.53 (SD=2325.62). The agreement between IFA and PCR in Walter Reed canine cells (WRCC) revealed a mean difference of -2396.55 with 95% confidence and an interval ranging from -3774.63 to -1018.48 (SD=2168.93). The Bland Altman test confirmed the overall accuracy of PCR vs IFA and the plot showed that all points were not randomly arranged in the range of average ± 1.96 × SD of the differences.

  2. Isothermal Titration Calorimetry: Assisted Crystallization of RNA-Ligand Complexes.

    PubMed

    Da Veiga, Cyrielle; Mezher, Joelle; Dumas, Philippe; Ennifar, Eric

    2016-01-01

    The success rate of nucleic acids/ligands co-crystallization can be significantly improved by performing preliminary biophysical analyses. Among suitable biophysical approaches, isothermal titration calorimetry (ITC) is certainly a method of choice. ITC can be used in a wide range of experimental conditions to monitor in real time the formation of the RNA- or DNA-ligand complex, with the advantage of providing in addition the complete binding profile of the interaction. Following the ITC experiment, the complex is ready to be concentrated for crystallization trials. This chapter describes a detailed experimental protocol for using ITC as a tool for monitoring RNA/small molecule binding, followed by co-crystallization.

  3. Characterization of binding interactions by isothermal titration calorimetry.

    PubMed

    Doyle

    1997-02-01

    Isothermal titration calorimetry is a high-accuracy method for measuring binding affinities. Titration calorimetry is a universal method that has broad impact throughout biotechnology. In recent years, microcalorimeters that are capable of characterizing binding interactions of biological macromolecules have become commercially available. Results from these studies are providing new insight into the molecular nature of macromolecular interactions.

  4. A computer-controlled potentiometric/spectrophotometric titrator

    PubMed Central

    Stong, John D.

    1988-01-01

    A laboratory computer controlled potentiometric titrator interfaced to a diode array spectrophotometer is described. The titrator consists of widely used, commercially available components; therefore, major attention is given to modes of interconnection and software implementation in data format and system control. Replicate potentiometric titrations of glycines gave a relative standard deviation in titre of 1.035% and a relative standard deviation in pH of 0.745%. Replicate spectrophotometric titrations of bromophenol blue were analysed at three wavelengths to yield pKa= 3.898 ± 0.075 (1.9% rsd). Methods of data presentation and manipulation are presented. PMID:18925194

  5. Methods for preparation of deuterated amino acids

    SciTech Connect

    Pshenichnikova, A.B.; Karnaukhova, E.N.; Zvonkova, E.N.

    1995-03-01

    The current state and prospects for the use of amino acids labeled with stable isotopes are considered. Methods for the preparation of deuterated amino acids, including synthetic, chemicoenzymatic, and biosynthetic ones, and deuterium exchange reactions are summarized. Problems in the preparation of optically pure amino acids are discussed. 120 refs., 15 figs.

  6. Safer staining method for acid fast bacilli.

    PubMed Central

    Ellis, R C; Zabrowarny, L A

    1993-01-01

    To develop a method for staining acid fast bacilli which excluded highly toxic phenol from the staining solution. A lipophilic agent, a liquid organic detergent, LOC High Studs, distributed by Amway, was substituted. The acid fast bacilli stained red; nuclei, cytoplasm, and cytoplasmic elements stained blue on a clear background. These results compare very favourably with acid fast bacilli stained by the traditional method. Detergents are efficient lipophilic agents and safer to handle than phenol. The method described here stains acid fast bacilli as efficiently as traditional carbol fuchsin methods. LOC High Suds is considerably cheaper than phenol. Images PMID:7687254

  7. Safer staining method for acid fast bacilli.

    PubMed

    Ellis, R C; Zabrowarny, L A

    1993-06-01

    To develop a method for staining acid fast bacilli which excluded highly toxic phenol from the staining solution. A lipophilic agent, a liquid organic detergent, LOC High Studs, distributed by Amway, was substituted. The acid fast bacilli stained red; nuclei, cytoplasm, and cytoplasmic elements stained blue on a clear background. These results compare very favourably with acid fast bacilli stained by the traditional method. Detergents are efficient lipophilic agents and safer to handle than phenol. The method described here stains acid fast bacilli as efficiently as traditional carbol fuchsin methods. LOC High Suds is considerably cheaper than phenol.

  8. Photometric method for determination of acidity constants through integral spectra analysis.

    PubMed

    Zevatskiy, Yuriy Eduardovich; Ruzanov, Daniil Olegovich; Samoylov, Denis Vladimirovich

    2015-04-15

    An express method for determination of acidity constants of organic acids, based on the analysis of the integral transmittance vs. pH dependence is developed. The integral value is registered as a photocurrent of photometric device simultaneously with potentiometric titration. The proposed method allows to obtain pKa using only simple and low-cost instrumentation. The optical part of the experimental setup has been optimized through the exclusion of the monochromator device. Thus it only takes 10-15 min to obtain one pKa value with the absolute error of less than 0.15 pH units. Application limitations and reliability of the method have been tested for a series of organic acids of various nature.

  9. Photometric method for determination of acidity constants through integral spectra analysis

    NASA Astrophysics Data System (ADS)

    Zevatskiy, Yuriy Eduardovich; Ruzanov, Daniil Olegovich; Samoylov, Denis Vladimirovich

    2015-04-01

    An express method for determination of acidity constants of organic acids, based on the analysis of the integral transmittance vs. pH dependence is developed. The integral value is registered as a photocurrent of photometric device simultaneously with potentiometric titration. The proposed method allows to obtain pKa using only simple and low-cost instrumentation. The optical part of the experimental setup has been optimized through the exclusion of the monochromator device. Thus it only takes 10-15 min to obtain one pKa value with the absolute error of less than 0.15 pH units. Application limitations and reliability of the method have been tested for a series of organic acids of various nature.

  10. A new sensor for thermometric titrations.

    PubMed

    Najib, Fadhil M; Zewar, Sardir; Abdulla, Ahmad M

    2007-01-15

    A new thermometric sensor, which is a transistor (OC71), has been introduced to follow thermometric titrations successfully to clear end points. The sensor was suitable in both normal and differential modes of titration. It is possible to titrate down to 1.32micromol of HCl and 26.4micromol of H(3)BO(3)in a final 20ml solution with accuracy and precision of 1%, 2.2% and 1.4%, 2.2%, respectively. The sensor, in association with a pH glass electrode, was used for the determination of pK values of some well established weak acids such as, acetic acid (4.77), phosphoric acid (pK(1)=2.18, pK(2)=7.20 and pK(3)=12.32) as well as for a very weak acid of uncertain pK values H(3)BO(3) (pK(1)=9.20, pK(2)=12.7 and pK(3)=13.80). The sensor was also examined for kinetic catalytic determination of iron(III) in water, milk and pharmaceuticals.

  11. Isothermal titration calorimetry of RNA.

    PubMed

    Salim, Nilshad N; Feig, Andrew L

    2009-03-01

    Isothermal titration calorimetry (ITC) is a fast and robust method to study the physical basis of molecular interactions. A single well-designed experiment can provide complete thermodynamic characterization of a binding reaction, including K(a), DeltaG, DeltaH, DeltaS and reaction stoichiometry (n). Repeating the experiment at different temperatures allows determination of the heat capacity change (DeltaC(P)) of the interaction. Modern calorimeters are sensitive enough to probe even weak biological interactions making ITC a very popular method among biochemists. Although ITC has been applied to protein studies for many years, it is becoming widely applicable in RNA biochemistry as well, especially in studies which involve RNA folding and RNA interactions with small molecules, proteins and with other RNAs. This review focuses on best practices for planning, designing and executing effective ITC experiments when one or more of the reactants is an RNA. PMID:18835447

  12. Well acidizing compositions and method

    SciTech Connect

    Gardener, T.R.; Dill, W.R.; Ford, W.G.F.; King, K.L.

    1991-07-23

    This patent describes a concentrate which forms an acid internal microemulsion well treatment composition when added to an acid treatment fluid. It comprises in the range of from about 20% to about 98% by weight of a hydrocarbon carrier fluid; in the range of from about 1% to about 50% by weight of an alkyl alcohol having in the range of from about 4 to 18 carbon atoms; and in the range of from about 1% to about 50% by weight of an emulsifying agent comprising at least one compound selected from the group consisting of amine salts having ester or amide linkages and propoxylated alcohols, each of the components being different compounds or different mixtures of compounds.

  13. Microscale pH Titrations Using an Automatic Pipet.

    ERIC Educational Resources Information Center

    Flint, Edward B.; Kortz, Carrie L.; Taylor, Max A.

    2002-01-01

    Presents a microscale pH titration technique that utilizes an automatic pipet. A small aliquot (1-5 mL) of the analyte solution is titrated with repeated additions of titrant, and the pH is determined after each delivery. The equivalence point is determined graphically by either the second derivative method or a Gran plot. The pipet can be…

  14. Rapid titration of viruses by flow cytometry.

    PubMed

    Drayman, Nir; Oppenheim, Ariella

    2011-06-01

    Traditionally, the most common methods used to titrate virus stocks are the plaque assay and the hemagglutination assay. The protocol presented here is based on the detection of viral-expressed proteins in infected cells by flow cytometry. It is simpler and more rapid than the traditional plaque-forming assay and it enables high-throughput analyses.

  15. Laboratory installation for the study of atomic-oxygen and ozone detectors and certain methodological aspects concerning the determination of oxygen-atom concentration by the methods of NO and NO2 titration

    NASA Astrophysics Data System (ADS)

    Bromberg, D. V.; Perov, S. P.

    A laboratory installation is described which can be used to study various characteristics of atomic oxygen and ozone in the pressure range from 0.01 to 50 Pa. The installation can be used to calibrate rocketborne sensors intended for measurements in the middle atmosphere. Systematic and random errors connected with the determination of oxygen-atom concentration by the NO2 and NO titration methods are examined.

  16. Estimation of avidin activity by two methods.

    PubMed

    Borza, B; Marcheş, F; Repanovici, R; Burducea, O; Popa, L M

    1991-01-01

    The biological activity of avidin was estimated by two different methods. The spectrophotometric method used the avidin titration with biotin in the presence of 4 hydroxiazobenzen-2'carboxilic acid as indicator. In the radioisotopic determination the titration with tritiated biotin was accomplished. Both methods led to the same results, but the spectrophotometric one is less avidin expensive and more rapid, being more convenient.

  17. An improved apparatus for intragastric titration in the conscious dog.

    PubMed

    Daly, M J; Hartley, R W; Stables, R

    1980-01-01

    An apparatus for automatic intragastric titration with on-line display of results has been described. The apparatus has been validated by titration of exogenous acid injected into the test meal in a reservoir and in vivo by assessing the secretory response of a dog with a gastric fistula to a test meal. A titration display unit provides a record of the secretory response both as a digital printout and a bar chart display. Cimetidine (2 mg kg-1h-1) significantly reduced the secretory response to a test meal.

  18. Proton-binding study of standard and reference fulvic acids, humic acids, and natural organic matter

    NASA Astrophysics Data System (ADS)

    Ritchie, Jason D.; Perdue, E. Michael

    2003-01-01

    The acid-base properties of 14 standard and reference materials from the International Humic Substances Society (IHSS) were investigated by potentiometric titration. Titrations were conducted in 0.1 M NaCl under a nitrogen atmosphere, averaging 30 min from start to finish. Concentrations of carboxyl groups and phenolic groups were estimated directly from titration curves. Titration data were also fit to a modified Henderson-Hasselbalch model for two classes of proton-binding sites to obtain "best fit" parameters that describe proton-binding curves for the samples. The model was chosen for its simplicity, its ease of implementation in computer spreadsheets, and its excellent ability to describe the shapes of the titration curves. The carboxyl contents of the IHSS samples are in the general order: terrestrial fulvic acids > aquatic fulvic acids > Suwannee River natural organic matter (NOM) > aquatic humic acids > terrestrial humic acids. Overall, fulvic acids and humic acids have similar phenolic contents; however, all of the aquatically derived samples have higher phenolic contents than the terrestrially derived samples. The acid-base properties of reference Suwannee River NOM are surprisingly similar to those of standard Suwannee River humic acid. Results from titrations in this study were compared with other published results from both direct and indirect titrations. Typically, carboxyl contents for the IHSS samples were in agreement with the results from both methods of titration. Phenolic contents for the IHSS samples were comparable to those determined by direct titrations, but were significantly less than estimates of phenolic content that were based on indirect titrations with Ba(OH) 2 and Ca(OAc) 2. The average phenolic-to-carboxylic ratio of the IHSS samples is approximately 1:4. Models that assume a 1:2 ratio of phenolic-to-carboxylic groups may overestimate the relative contribution of phenolic groups to the acid-base chemistry of humic substances.

  19. Novel approaches to analysis by flow injection gradient titration.

    PubMed

    Wójtowicz, Marzena; Kozak, Joanna; Kościelniak, Paweł

    2007-09-26

    Two novel procedures for flow injection gradient titration with the use of a single stock standard solution are proposed. In the multi-point single-line (MP-SL) method the calibration graph is constructed on the basis of a set of standard solutions, which are generated in a standard reservoir and subsequently injected into the titrant. According to the single-point multi-line (SP-ML) procedure the standard solution and a sample are injected into the titrant stream from four loops of different capacities, hence four calibration graphs are able to be constructed and the analytical result is calculated on the basis of a generalized slope of these graphs. Both approaches have been tested on the example of spectrophotometric acid-base titration of hydrochloric and acetic acids with using bromothymol blue and phenolphthalein as indicators, respectively, and sodium hydroxide as a titrant. Under optimized experimental conditions the analytical results of precision less than 1.8 and 2.5% (RSD) and of accuracy less than 3.0 and 5.4% (relative error (RE)) were obtained for MP-SL and SP-ML procedures, respectively, in ranges of 0.0031-0.0631 mol L(-1) for samples of hydrochloric acid and of 0.1680-1.7600 mol L(-1) for samples of acetic acid. The feasibility of both methods was illustrated by applying them to the total acidity determination in vinegar samples with precision lower than 0.5 and 2.9% (RSD) for MP-SL and SP-ML procedures, respectively.

  20. Method for sequencing nucleic acid molecules

    DOEpatents

    Korlach, Jonas; Webb, Watt W.; Levene, Michael; Turner, Stephen; Craighead, Harold G.; Foquet, Mathieu

    2006-06-06

    The present invention is directed to a method of sequencing a target nucleic acid molecule having a plurality of bases. In its principle, the temporal order of base additions during the polymerization reaction is measured on a molecule of nucleic acid, i.e. the activity of a nucleic acid polymerizing enzyme on the template nucleic acid molecule to be sequenced is followed in real time. The sequence is deduced by identifying which base is being incorporated into the growing complementary strand of the target nucleic acid by the catalytic activity of the nucleic acid polymerizing enzyme at each step in the sequence of base additions. A polymerase on the target nucleic acid molecule complex is provided in a position suitable to move along the target nucleic acid molecule and extend the oligonucleotide primer at an active site. A plurality of labelled types of nucleotide analogs are provided proximate to the active site, with each distinguishable type of nucleotide analog being complementary to a different nucleotide in the target nucleic acid sequence. The growing nucleic acid strand is extended by using the polymerase to add a nucleotide analog to the nucleic acid strand at the active site, where the nucleotide analog being added is complementary to the nucleotide of the target nucleic acid at the active site. The nucleotide analog added to the oligonucleotide primer as a result of the polymerizing step is identified. The steps of providing labelled nucleotide analogs, polymerizing the growing nucleic acid strand, and identifying the added nucleotide analog are repeated so that the nucleic acid strand is further extended and the sequence of the target nucleic acid is determined.

  1. Method for sequencing nucleic acid molecules

    DOEpatents

    Korlach, Jonas; Webb, Watt W.; Levene, Michael; Turner, Stephen; Craighead, Harold G.; Foquet, Mathieu

    2006-05-30

    The present invention is directed to a method of sequencing a target nucleic acid molecule having a plurality of bases. In its principle, the temporal order of base additions during the polymerization reaction is measured on a molecule of nucleic acid, i.e. the activity of a nucleic acid polymerizing enzyme on the template nucleic acid molecule to be sequenced is followed in real time. The sequence is deduced by identifying which base is being incorporated into the growing complementary strand of the target nucleic acid by the catalytic activity of the nucleic acid polymerizing enzyme at each step in the sequence of base additions. A polymerase on the target nucleic acid molecule complex is provided in a position suitable to move along the target nucleic acid molecule and extend the oligonucleotide primer at an active site. A plurality of labelled types of nucleotide analogs are provided proximate to the active site, with each distinguishable type of nucleotide analog being complementary to a different nucleotide in the target nucleic acid sequence. The growing nucleic acid strand is extended by using the polymerase to add a nucleotide analog to the nucleic acid strand at the active site, where the nucleotide analog being added is complementary to the nucleotide of the target nucleic acid at the active site. The nucleotide analog added to the oligonucleotide primer as a result of the polymerizing step is identified. The steps of providing labelled nucleotide analogs, polymerizing the growing nucleic acid strand, and identifying the added nucleotide analog are repeated so that the nucleic acid strand is further extended and the sequence of the target nucleic acid is determined.

  2. Electrochemical titrations of thiosulfate, sulfite, dichromate and permanganate using dual microband electrodes.

    PubMed

    Rajantie, H; Williams, D E

    2001-01-01

    Applications of titration using dual gold microband electrodes in generator-collector mode are presented. The main advantage is that the method dispenses with the need for accurate volume measurements and reagent preparation: rather than balancing of molar amounts, the fluxes of analyte and titrant are balanced instead. The titrant is generated electrochemically and the end-point is detected amperometrically by the appearance of a current due to the presence of the titrant at the other band. The aim is to show the versatility and capability of the method that employs disposable mass-produced electrodes and uses pulsed motion of the electrode before a measurement to renew the boundary conditions. The titration of ascorbic acid (vitamin C) with ferricyanide, which has been a model system, was extended to the determination of thiosulfate and sulfite with iodine and the determination of dichromate and permanganate with iron(II). The accuracy, limited by the present fabrication reproducibility of the disposable electrodes, is +/- 10%. The results demonstrate that the method is effective even with a very simple set-up, and the actual time needed for common titrations is significantly decreased. These examples demonstrate a comprehensive basis for further development and applications, including standard titrations in industry.

  3. Theoretical study of the acid-base properties of the montmorillonite/electrolyte interface: influence of the surface heterogeneity and ionic strength on the potentiometric titration curves.

    PubMed

    Zarzycki, Piotr; Thomas, Fabien

    2006-10-15

    The parallel shape of the potentiometric titration curves for montmorillonite suspension is explained using the surface complexation model and taking into account the surface heterogeneity. The homogeneous models give accurate predictions only if they assume unphysically large values of the equilibrium constants for the exchange process occurring on the basal plane. However, the assumption that the basal plane is energetically heterogeneous allows to fit the experimental data (reported by Avena and De Pauli [M. Avena, C.P. De Pauli, J. Colloid Interface Sci. 202 (1998) 195-204]) for reasonable values of exchange equilibrium constant equal to 1.26 (suggested by Fletcher and Sposito [P. Fletcher, G. Sposito, Clay Miner. 24 (1989) 375-391]). Moreover, we observed the typical behavior of point of zero net proton charge (pznpc) as a function of logarithm of the electrolyte concentration (log[C]). We showed that the slope of the linear dependence, pznpc=f(log[C]), is proportional to the number of isomorphic substitutions in the crystal phase, which was also observed in the experimental studies.

  4. Protometric thermometric titrations of sparingly soluble compounds in water in the presence of n-octanol.

    PubMed

    Burgot, G; Burgot, J-L

    2002-10-15

    Thermometric titrimetry permits titration of acido-basic compounds in water in the presence of n-octanol. n-Octanol permits the solubilization of protolytes and moreover may also displace the equilibria of the titration reactions. Hydrochlorides of highly insoluble derivatives such as phenothiazine derivatives can be titrated with satisfactory accuracy and precision by sodium hydroxide despite their high pK(a) values. Likewise barbiturate salts can be titrated by hydrochloric acid. In the case of some salts, the methodology may permit the sequential titration of the ion and counter ion.

  5. A novel view of pH titration in biomolecules.

    PubMed

    Onufriev, A; Case, D A; Ullmann, G M

    2001-03-27

    When individual titratable sites in a molecule interact with each other, their pH titration can be considerably more complex than that of an independent site described by the classical Henderson-Hasselbalch equation. We propose a novel framework that decomposes any complex titration behavior into simple standard components. The approach maps the set of N interacting sites in the molecule onto a set of N independent, noninteracting quasi-sites, each characterized by a pK'(a) value. The titration curve of an individual site in the molecule is a weighted sum of Henderson-Hasselbalch curves corresponding to the quasi-sites. The total protonation curve is the unweighted sum of these Henderson-Hasselbalch curves. We show that pK'(a) values correspond to deprotonation constants available from methods that can be used to assess total proton uptake or release, and establish their connection to protonation curves of individual residues obtained by NMR or infrared spectroscopy. The new framework is tested on a small molecule diethylenetriaminepentaacetate (DTPA) exhibiting nonmonotonic titration curves, where it gives an excellent fit to experimental data. We demonstrate that the titration curve of a site in a group of interacting sites can be accurately reconstructed, if titration curves of the other sites are known. The application of the new framework to the protein rubredoxin demonstrates its usefulness in calculating and interpreting complicated titration curves.

  6. Analysis of variance in determinations of equivalence volume and of the ionic product of water in potentiometric titrations.

    PubMed

    Braibanti, A; Bruschi, C; Fisicaro, E; Pasquali, M

    1986-06-01

    Homogeneous sets of data from strong acid-strong base potentiometric titrations in aqueous solution at various constant ionic strengths have been analysed by statistical criteria. The aim is to see whether the error distribution matches that for the equilibrium constants determined by competitive potentiometric methods using the glass electrode. The titration curve can be defined when the estimated equivalence volume VEM, with standard deviation (s.d.) sigma (VEM), the standard potential E(0), with s.d. sigma(E(0)), and the operational ionic product of water K(*)(w) (or E(*)(w) in mV), with s.d. sigma(K(*)(w)) [or sigma(E(*)(w))] are known. A special computer program, BEATRIX, has been written which optimizes the values of VEM, E(0) and K(*)(w) by linearization of the titration curve as a Gran plot. Analysis of variance applied to a set of 11 titrations in 1.0M sodium chloride medium at 298 K has demonstrated that the values of VEM belong to a normal population of points corresponding to individual potential/volume data-pairs (E(i); v(i)) of any titration, whereas the values of pK(*)(w) (or of E(*)(w)) belong to a normal population with members corresponding to individual titrations, which is also the case for the equilibrium constants. The intertitration variation is attributable to the electrochemical component of the system and appears as signal noise distributed over the titrations. The correction for junction-potentials, introduced in a further stage of the program by optimization in a Nernst equation, increases the noise, i.e., sigma(pK(*)(w)). This correction should therefore be avoided whenever it causes an increase of sigma(pK(*)(w)). The influence of the ionic medium has been examined by processing data from acid-base titrations in 0.1M potassium chloride and 0.5M potassium nitrate media. The titrations in potassium chloride medium showed the same behaviour as those in sodium chloride medium, but with an s.d. for pK(*)(w) that was smaller and close to the

  7. Challenges with fats and fatty acid methods.

    PubMed

    Palmquist, D L; Jenkins, T C

    2003-12-01

    The content and chemical nature of lipids in feedstuffs is heterogeneous. It has long been known that ether extraction by the Weende procedure inadequately characterizes the fat content of feedstuffs, yet it remains the official method. Diethyl ether (or hexanes that are often used) extracts significant amounts of nonnutritive, nonsaponifiable lipids from forages, and often incompletely extracts lipids of nutritional value, especially fatty acids present as salts of divalent cations. Preextraction hydrolysis of insoluble fatty acid salts with acid releases these fatty acids, and this step is included in the official procedure for certain feedstuffs in the United Kingdom; however, acid hydrolysis increases analysis time and decreases precision. Acid hydrolysis also causes confusion as to the proper definition of the fat content of feedstuffs. A preferred method of fat analysis determines the total fatty acid concentration in feed samples by converting fatty acid salts, as well as the acyl components in all lipid classes, such as triacylglycerols, phospholipids, and sphingolipids, to methyl esters using a simple, direct one-step esterification procedure. Fatty acid methyl esters are then quantified by GLC, which provides information on both fatty acid quantity and profile in a single analysis. Adjustments in conditions and reagents may be necessary to overcome difficulty in quantitatively preparing esters from certain types of fatty acids and their derivatives in commercial fat supplements. After correction for glycerol content, analysis of oils by this procedure provides information on the content of nonsaponifiable material, such as chlorophyll, waxes, and indigestible polymers formed from heat- or oxidatively damaged fats. The correct description of feedstuffs for energy value of fats is the content of total fatty acids. PMID:14677882

  8. Purity determination and uncertainty evaluation of folic acid by mass balance method.

    PubMed

    Gong, Hui; Huang, Ting; Yang, Yi; Wang, Haifeng

    2012-11-15

    Folic acid is one of the most important nutrient substances for human beings, especially for the pregnant women and infants. Therefore the purity determination of folic acid is particularly important. The mass balance method was employed to determine the purity of folic acid, by using the measures of high performance liquid chromatography (HPLC), Karl Fischer titration and other conventional approach. The moisture quantification of folic acid was a major problem since it is a thermally unstable substance and it is apt to contain crystal water. Therefore, a novel improved Karl Fischer method was established for accurate determination of the water content in folic acid, whose repeatability (RSD=2.9%) was significantly better than that of the original direct injection method (RSD=12%). The purity of folic acid certified reference material (CRM) determined by mass balance method was 90.9% with an expanded uncertainty of 0.35%, and the content of water (the major impurity) was 8.5%, with an expanded uncertainty of 0.32%.

  9. Automatic Titrators in the Analytical and Physical Chemistry Laboratories

    NASA Astrophysics Data System (ADS)

    Williams, Kathryn R.

    1998-09-01

    In 1995 the University of Florida received an NSF-ILI grant to purchase six automatic titrators, which have now been successfully integrated into the analytical and physical chemistry teaching laboratories. After they have mastered fundamental techniques, students in the introductory analytical laboratory gain experience with automated analyses in three experiments: the iodimetric analysis of ascorbic acid, the determination of polymer molecular weight, and the analysis of chloride by ion selective electrode. The titrators are also used in the instrumental analysis laboratory for Karl Fischer titrations and the coulometric analysis of EDTA. A physical chemistry experiment, the kinetics of methyl acetate hydrolysis, has also been modified for use with the titrators Instructor write-ups for all experiments may be obtained via JCE Online.

  10. Methods for analyzing nucleic acid sequences

    DOEpatents

    Korlach, Jonas; Webb, Watt W.; Levene, Michael; Turner, Stephen; Craighead, Harold G.; Foquet, Mathieu

    2011-05-17

    The present invention is directed to a method of sequencing a target nucleic acid. The method provides a complex comprising a polymerase enzyme, a target nucleic acid molecule, and a primer, wherein the complex is immobilized on a support Fluorescent label is attached to a terminal phosphate group of the nucleotide or nucleotide analog. The growing nucleic acid strand is extended by using the polymerase to add a nucleotide analog to the nucleic acid strand. The nucleotide analog added to the oligonucleotide primer as a result of the polymerizing step is identified. The time duration of the signal from labeled nucleotides or nucleotide analogs that become incorporated is distinguished from freely diffusing labels by a longer retention in the observation volume for the nucleotides or nucleotide analogs that become incorporated than for the freely diffusing labels.

  11. Phenolic acid esterases, coding sequences and methods

    DOEpatents

    Blum, David L.; Kataeva, Irina; Li, Xin-Liang; Ljungdahl, Lars G.

    2002-01-01

    Described herein are four phenolic acid esterases, three of which correspond to domains of previously unknown function within bacterial xylanases, from XynY and XynZ of Clostridium thermocellum and from a xylanase of Ruminococcus. The fourth specifically exemplified xylanase is a protein encoded within the genome of Orpinomyces PC-2. The amino acids of these polypeptides and nucleotide sequences encoding them are provided. Recombinant host cells, expression vectors and methods for the recombinant production of phenolic acid esterases are also provided.

  12. Surface Titrations of Perlite Suspensions.

    PubMed

    Alkan; Do

    1998-11-01

    The surface charge behaviour of unexpanded and expanded perlite samples in KNO3 and NaCl solutions were investigated as a function of pH and ionic strength. The solutions of KNO3 and NaCl ranging from 10(-3) to 1.0 M were used. The potentiometric titration method was used to determine the surface charge of perlite samples. It was confirmed that the perlite samples had no the point of zero charge and was negatively charged in the pH range of 3-10. The double extrapolation method was used for determining the intrinsic equilibrium constants for simple ionization and complex ionization reactions. The values obtained are pKinta2 = 2.5 and p*KintK+ = 2.3 in KNO3 solutions and pKinta2 = 3.0 and p*KintNa+ = 2.4 in NaCl solutions for unexpanded perlite, and pKinta2 = 2.6 and p*KintK+ = 2.4 in KNO3 solutions and pKinta2 = 2.7 and pKintNa+ = 2.4 in NaCl solutions for expanded perlite. Copyright 1998 Academic Press.

  13. Automatic photometric titrations of calcium and magnesium in carbonate rocks

    USGS Publications Warehouse

    Shapiro, L.; Brannock, W.W.

    1955-01-01

    Rapid nonsubjective methods have been developed for the determination of calcium and magnesium in carbonate rocks. From a single solution of the sample, calcium is titrated directly, and magnesium is titrated after a rapid removal of R2O3 and precipitation of calcium as the tungstate. A concentrated and a dilute solution of disodium ethylenediamine tetraacetate are used as titrants. The concentrated solution is added almost to the end point, then the weak solution is added in an automatic titrator to determine the end point precisely.

  14. Continuous on-line true titrations by feedback-based flow ratiometry. The principle of compensating errors

    PubMed

    Tanaka; Dasgupta; Huang

    2000-10-01

    We introduce a new concept for continuous on-line titrations based on feedback-controlled flow ratiometry and the principle of compensating errors. The system has been thoroughly tested by applying it to acid-base neutralization titrations with indicator-based end point detection. In a typical case, the total flow (FT, consisting of the sample and the titrant flows) is held constant while the titrant (e.g., a standard base containing an indicator) flow FB varies linearly in response to a controller output voltage. The sample (e.g., an acidic solution to be titrated) flow FA constitutes the makeup and thus also varies (FA = FT - FB). The status of the indicator color in the mixed stream is monitored by an optical detector and used either for governing the controller output or for interpreting the results of the titration. Three methods (PID based control, fixed triangular wave control, and feedback-based triangular wave control implemented on a PC) were examined. In the last and the most successful approach, the titrant flow is initially ramped upward linearly. At the instant a change in the color is sensed by the detector, the titrant flow rate FH is higher than the true equivalence flow rate FE because of the lag time between the first compositional change and its detection. The sensing of the change in color causes the system output to immediately reverse its ramp direction such that the titrant flow now goes down linearly at the same rate. At the instant a change in color, in the opposite direction this time, is again sensed, the titrant flow rate FL is lower than FE by exactly the same amount that FH was higher than FE. This principle of compensating errors (FE = (FH + FL)/2) allows true titrations with excellent reproducibility and speed (0.6% RSD at 3 s/titration and 0.2% RSD at 10 s/titration) and titrant volume consumption as little as 12 microL/titration and solves an old conceptual problem in flow based titrations.

  15. Logarithmic diagrams as an aid in complexometric titrations where 1:2 or 2:1 as well as 1:1 metal-ligand complexes are regarded as principal species.

    PubMed

    Johansson, S

    1981-04-01

    Logarithmic diagrams have earlier been used for theoretical treatment of complexometric titrations where only the 1:1 complex is formed in the main reaction between a metal ion M and a ligand L. This approach has now been extended to include titrations where 1:2 and 2:1 complexes are also formed. Four different types of diagram can be drawn. These diagrams can be used to determine pM (or pL) at the equivalence point, the titration ratio at different pM (or pL) values and the titration error. One of these diagrams is easy to construct and use. The method also has the advantage that it can directly indicate whether the titration is best performed with 1:1 or 2:1 (1:2) stoichiometry. Two typical examples, namely the titration of a manganese solution with triethylenetetraminehexa-acetic acid and of a copper solution with pyridine-2,6-dicarboxylic acid, are discussed. The two titrations proceed with the formation of the complexes Mn(2)L (2: 1) and CuL(2) (1:2) respectively, in addition to the 1:1 complex in each instance.

  16. Some new applications of ferroin as redox indicator in titrations with dichromate.

    PubMed

    Sriramam, K

    1972-11-01

    Working conditions for the titration of arsenic(III), hydroquinone, ferrocyanide, uranium(IV) and molybdenum(V) with dichromate in sulphuric acid and hydrochloric acid media have been established, with ferroin as the redox indicator.

  17. Isothermal Titration Calorimetry: Assisted Crystallization of RNA-Ligand Complexes.

    PubMed

    Da Veiga, Cyrielle; Mezher, Joelle; Dumas, Philippe; Ennifar, Eric

    2016-01-01

    The success rate of nucleic acids/ligands co-crystallization can be significantly improved by performing preliminary biophysical analyses. Among suitable biophysical approaches, isothermal titration calorimetry (ITC) is certainly a method of choice. ITC can be used in a wide range of experimental conditions to monitor in real time the formation of the RNA- or DNA-ligand complex, with the advantage of providing in addition the complete binding profile of the interaction. Following the ITC experiment, the complex is ready to be concentrated for crystallization trials. This chapter describes a detailed experimental protocol for using ITC as a tool for monitoring RNA/small molecule binding, followed by co-crystallization. PMID:26227041

  18. Studying Current-Potential Curves Using a Bipotentiometric Lodometric Back-Titration for the Determination of Ascorbic Acid in Fruits and Vegetables

    ERIC Educational Resources Information Center

    Verdini, Roxana A.; Lagier, Claudia M.

    2004-01-01

    Voltammetry principles are introduced to students by means of a bipotentiometric method to determine vitamin C in fruits and vegetables. The aim is to draw attention to voltammetric methods, particular to the study of current-potential curves, stressing the potential applicability in areas of food quality control.

  19. Measuring titratable alkalinity by single versus double endpoint titration: An evaluation in two cyprinodont species and implications for characterizing net H+ flux in aquatic organisms.

    PubMed

    Brix, Kevin V; Wood, Chris M; Grosell, Martin

    2013-01-01

    In this study, Na(+) uptake and acid-base balance in the euryhaline pupfish Cyprinodon variegatus variegatus were characterized when fish were exposed to pH 4.5 freshwater (7mM Na(+)). Similar to the related cyprinodont, Fundulus heteroclitus, Na(+) uptake was significantly inhibited when exposed to low pH water. However, it initially appeared that C. v. variegatus increased apparent net acid excretion at low pH relative to circumneutral pH. This result is opposite to previous observations for F. heteroclitus under similar conditions where fish were observed to switch from apparent net H(+) excretion at circumneutral pH to apparent net H(+) uptake at low pH. Further investigation revealed disparate observations between these studies were the result of using double endpoint titrations to measure titratable alkalinity fluxes in the current study, while the earlier study utilized single endpoint titrations to measure these fluxes (i.e.,. Cyprinodon acid-base transport is qualitatively similar to Fundulus when characterized using single endpoint titrations). This led to a comparative investigation of these two methods. We hypothesized that either the single endpoint methodology was being influenced by a change in the buffer capacity of the water (e.g., mucus being released by the fish) at low pH, or the double endpoint methodology was not properly accounting for ammonia flux by the fish. A series of follow-up experiments indicated that buffer capacity of the water did not change significantly, that excretion of protein (a surrogate for mucus) was actually reduced at low pH, and that the double endpoint methodology does not properly account for NH(3) excretion by fish under low pH conditions. As a result, it overestimates net H(+) excretion during low pH exposure. After applying the maximum possible correction for this error (i.e., assuming that all ammonia is excreted as NH(3)), the double endpoint methodology indicates that net H(+) transport was reduced to

  20. Determination of dissociation parameters of weak acids in different media according to the isohydric method.

    PubMed

    Michałowski, Tadeusz; Pilarski, Bogusław; Asuero, Agustin G; Dobkowska, Agnieszka; Wybraniec, Sławomir

    2011-10-30

    The isohydricity (pH constancy) principle is referred to the pair of solutions: weak acid (HL, C(0)mol/L) and strong acid (HB, C mol/L) when mixed e.g., according to titrimetric mode. Such a case takes place if the relation C(0)=C+C(2) × 10(pK(1)) is valid, where pK(1)=-log K(1), K(1) - dissociation constant for a weak monoprotic acid HL. This principle, outlined and formulated in earlier paper (Michałowski et al., Talanta 82 (2010) 1965), is the basis for a sensitive method of pK(1) determination, confirmed for a series of weak acids in presence of basal electrolytes or in water+organic solvent (dimethyl sulphoxide, methanol, isopropanol) media. The results of titrations were elaborated according to principles of regression analysis, with use of least squares method. A new criterion for precision of the results obtained according to this method is formulated. The pK(1) values obtained are comparable with ones found in literature.

  1. A Laser-Pointer-Based Spectrometer for Endpoint Detection of EDTA Titrations

    ERIC Educational Resources Information Center

    Dahm, Christopher E.; Hall, James W.; Mattioni, Brian E.

    2004-01-01

    A laser spectrometer for the ethylenediaminetetra-acetic acid (EDTA) titration of magnesium or calcium ions that is designed around a handheld laser pointer as the source and a photoresistor as the detector is developed. Findings show that the use of the spectrometer reduces the degree of uncertainty and error in one part of the EDTA titrations,…

  2. Method for the production of dicarboxylic acids

    DOEpatents

    Nghiem, N.P.; Donnelly, M.; Millard, C.S.; Stols, L.

    1999-02-09

    The present invention is an economical fermentation method for the production of carboxylic acids comprising the steps of (a) inoculating a medium having a carbon source with a carboxylic acid-producing organism; (b) incubating the carboxylic acid-producing organism in an aerobic atmosphere to promote rapid growth of the organism thereby increasing the biomass of the organism; (c) controllably releasing oxygen to maintain the aerobic atmosphere; (d) controllably feeding the organism having increased biomass with a solution containing the carbon source to maintain the concentration of the carbon source within the medium of about 0.5 g/l up to about 1 g/l; (e) depriving the aerobic atmosphere of oxygen to produce an anaerobic atmosphere to cause the organism to undergo anaerobic metabolism; (f) controllably feeding the organism having increased biomass a solution containing the carbon source to maintain the concentration of the carbon source within the medium of {>=}1 g/l; and (g) converting the carbon source to carboxylic acids using the anaerobic metabolism of the organism. 7 figs.

  3. Method for the production of dicarboxylic acids

    DOEpatents

    Nghiem, Nhuan Phu; Donnelly, Mark; Millard, Cynthia S.; Stols, Lucy

    1999-01-01

    The present invention is an economical fermentation method for the production of carboxylic acids comprising the steps of a) inoculating a medium having a carbon source with a carboxylic acid-producing organism; b) incubating the carboxylic acid-producing organism in an aerobic atmosphere to promote rapid growth of the organism thereby increasing the biomass of the organism; c) controllably releasing oxygen to maintain the aerobic atmosphere; d) controllably feeding the organism having increased biomass with a solution containing the carbon source to maintain the concentration of the carbon source within the medium of about 0.5 g/L up to about 1 g/L; e) depriving the aerobic atmosphere of oxygen to produce an anaerobic atmosphere to cause the organism to undergo anaerobic metabolism; f) controllably feeding the organism having increased biomass a solution containing the carbon source to maintain the concentration of the carbon source within the medium of .gtoreq.1 g/L; and g) converting the carbon source to carboxylic acids using the anaerobic metabolism of the organism.

  4. Analyzing a kinetic titration series using affinity biosensors.

    PubMed

    Karlsson, Robert; Katsamba, Phinikoula S; Nordin, Helena; Pol, Ewa; Myszka, David G

    2006-02-01

    The classical method of measuring binding constants with affinity-based biosensors involves testing several analyte concentrations over the same ligand surface and regenerating the surface between binding cycles. Here we describe an alternative approach to collecting kinetic binding data, which we call "kinetic titration." This method involves sequentially injecting an analyte concentration series without any regeneration steps. Through a combination of simulation and experimentation, we show that this method can be as robust as the classical method of analysis. In addition, kinetic titrations can be more efficient than the conventional data collection method and allow us to fully characterize analyte binding to ligand surfaces that are difficult to regenerate.

  5. Kinetics of reactions of antihemagglutinin and antineuraminidase antibodies with H2N2 and H3N2 influenza virus strains and description of a modification of the photometric ACU method for titration of antineuraminidase antibodies.

    PubMed

    Drescher, J; Desselberger, U

    1976-01-01

    The isotherms describing the reactions of selected H2N2 and H3N2 virus strains with antihemagglutinin (AH) and antineuraminidase (AN) antibodies were established by use of a photometric hemagglutination inhibition test [antibody concentration unit (ACU) method]. It was found that the AN antibody isotherms had significantly higher values of the constant 1/N than did the AH antibody isotherms. This finding confirms for further virus strains the conclusion that the photometric ACU method can discriminate objectively between AN and AH antibodies. The results obtained when determining by use of the photometric ACU method the kinetics of reactions of AN antibodies oriented to A/Bel (HO)-A/Sing (N2) virus with the neuraminidases of H2N2 and H3N2 strains, and vice versa, indicated that the N2 neuraminidases of the test strains could be divided into the following groups: One group comprising the strains A/Sing/1/57 and A/AA/1/65, a second comprising the strains A/Hong Kong/1/68 and A/England/42/72 and a third represented by A/Port Chalmers/1/73. This finding indicated progressive antigenic variation of the neuraminidases of the strains tested. A modification of the photometric ACU method for the titration of AN antibodies oriented to N2 strains has been developed. The modified technique was found to be more sensitive and accurate than was AN antibody titration by means of enzyme inhibition and HI pattern test.

  6. A Geochemical Reaction Model for Titration of Contaminated Soil and Groundwater at the Oak Ridge Reservation

    NASA Astrophysics Data System (ADS)

    Zhang, F.; Parker, J. C.; Gu, B.; Luo, W.; Brooks, S. C.; Spalding, B. P.; Jardine, P. M.; Watson, D. B.

    2007-12-01

    This study investigates geochemical reactions during titration of contaminated soil and groundwater at the Oak Ridge Reservation in eastern Tennessee. The soils and groundwater exhibits low pH and high concentrations of aluminum, calcium, magnesium, manganese, various trace metals such as nickel and cobalt, and radionuclides such as uranium and technetium. The mobility of many of the contaminant species diminishes with increasing pH. However, base additions to increase pH are strongly buffered by various precipitation/dissolution and adsorption/desorption reactions. The ability to predict acid-base behavior and associated geochemical effects is thus critical to evaluate remediation performance of pH manipulation strategies. This study was undertaken to develop a practical but generally applicable geochemical model to predict aqueous and solid-phase speciation during soil and groundwater titration. To model titration in the presence of aquifer solids, an approach proposed by Spalding and Spalding (2001) was utilized, which treats aquifer solids as a polyprotic acid. Previous studies have shown that Fe and Al-oxyhydroxides strongly sorb dissolved Ni, U and Tc species. In this study, since the total Fe concentration is much smaller than that of Al, only ion exchange reactions associated with Al hydroxides are considered. An equilibrium reaction model that includes aqueous complexation, precipitation, ion exchange, and soil buffering reactions was developed and implemented in the code HydroGeoChem 5.0 (HGC5). Comparison of model results with experimental titration curves for contaminated groundwater alone and for soil- water systems indicated close agreement. This study is expected to facilitate field-scale modeling of geochemical processes under conditions with highly variable pH to develop practical methods to control contaminant mobility at geochemically complex sites.

  7. Complexometric titrations: new reagents and concepts to overcome old limitations.

    PubMed

    Zhai, Jingying; Bakker, Eric

    2016-07-21

    Chelators and end point indicators are the most important parts of complexometric titrations. The most widely used universal chelator ethylenediamine tetraacetic acid (EDTA) and its derivatives can strongly coordinate with different metal ions. Their limited selectivity often requires the use of masking agents, and the multiple pKa values of the chelators necessitate a careful adjustment of pH during the procedure. Real world requirements for pH independent, selective and sensitive chelators and indicators call for a new design of these reagents. New concepts and structures of chelators and indicators have indeed recently emerged. We present here recent developments on chelators and indicators for complexometric titrations. Many of these advances were made possible only recently by moving the titration from a homogeneous to a heterogeneous phase using a new class of chelators and indicators based on highly selective ionophores embedded in ion-selective nanosphere emulsions. In view of achieving titrations in situ by complete instrumental control, thin layer electrochemistry has recently been shown to be an attractive concept that replaces the traditional cumbersome titration protocol with a direct reagent free sensing tool. PMID:27272695

  8. Surface characterization of hydroxyapatite: potentiometric titrations coupled with solubility measurements.

    PubMed

    Skartsila, Kyriaki; Spanos, Nikos

    2007-04-15

    The acid-base properties of synthetically prepared and well-characterized hydroxyapatite (HAP) in contact with KNO3 solutions were investigated at 25 degrees C, through potentiometric titrations, zeta-potential measurements, and surface complex modeling. Aliquots of suspension were withdrawn every 0.5 pH unit during the titration procedure and analyzed for calcium and phosphate. It was found that, even for rapid titration experiments, a remarkable amount of H+ ions (H+dissol.) is consumed in the bulk solution in reacting with species coming from the dissolution of HAP. These H+ ions must be taken into account in the H+ mass balance, in order for true value for the point of zero charge (pzc=6.5+/-0.2) and consequently true value for the surface charge (sigma0) to be obtained. Besides the conventional potentiometric titration technique, it was found that pzc may be determined much more easily as the intersection point of the suspension titration curve and the blank one modified to include the amount of H+dissol. obtained at one ionic strength. Finally, a surface complexation model was proposed for the development of surface charge. Experimental data were satisfactorily fitted by using the value of 4.2 F m-2 for the capacitance.

  9. Complexometric titrations: new reagents and concepts to overcome old limitations.

    PubMed

    Zhai, Jingying; Bakker, Eric

    2016-07-21

    Chelators and end point indicators are the most important parts of complexometric titrations. The most widely used universal chelator ethylenediamine tetraacetic acid (EDTA) and its derivatives can strongly coordinate with different metal ions. Their limited selectivity often requires the use of masking agents, and the multiple pKa values of the chelators necessitate a careful adjustment of pH during the procedure. Real world requirements for pH independent, selective and sensitive chelators and indicators call for a new design of these reagents. New concepts and structures of chelators and indicators have indeed recently emerged. We present here recent developments on chelators and indicators for complexometric titrations. Many of these advances were made possible only recently by moving the titration from a homogeneous to a heterogeneous phase using a new class of chelators and indicators based on highly selective ionophores embedded in ion-selective nanosphere emulsions. In view of achieving titrations in situ by complete instrumental control, thin layer electrochemistry has recently been shown to be an attractive concept that replaces the traditional cumbersome titration protocol with a direct reagent free sensing tool.

  10. Precise chelatometric titrations of zinc, cadmium, and lead with molecular spectroscopy.

    PubMed

    Suzuki, Toshihiro; Tiwari, Diwakar; Hioki, Akiharu

    2007-10-01

    Spectral data during a chelatometric titration were utilized for determining an equivalence point of the titration. Three-dimensional titration data were analyzed by fitting to a theoretical equation that was derived from equilibrium equations among a metal ion, an indicator and EDTA. The equation with optimized parameters successfully explained the three-dimensional titration data. This method was applied to EDTA titration systems of zinc, cadmium and lead with xylenol orange (XO) as an indicator. These systems were analyzed while taking the formation of 1:1 and 2:1 metal-indicator complexes into account. By this method, an accurate equivalence point was determined as well as stability constants and molar absorption spectra of metal-indicator complexes in each titration system. Moreover, the comparability among the concentrations of the metal standard solutions prepared from pure metals (Zn, Pb, and Cd) was also confirmed by the proposed evaluation method.

  11. Tracer monitored titrations: measurement of dissolved oxygen.

    PubMed

    Martz, Todd; Takeshita, Yuichiro; Rolph, Rebecca; Bresnahan, Philip

    2012-01-01

    The tracer monitored titration (TMT) technique is evaluated for measurement of dissolved oxygen. The TMT developed in this work uses a simple apparatus consisting of a low-precision pump for titrant delivery and an optical detector based on a white LED and two photodiodes with interference filters. It is shown that the classic Winkler method can be made free of routine volumetric and gravimetric measurements by application of TMT theory, which allows tracking the amounts of titrant and sample using a chemical tracer. The measurement precision of the prototype setup was 0.3% RSD.

  12. Determination of lanthanum by flame photometric titration.

    PubMed

    Svehla, G; Slevin, P J

    1968-09-01

    The flame emission of lanthanum at 560 mmu decreases linearly with phosphate concentration until a 1:1 molar ratio is reached, and then remains practically constant. Lanthanum can be titrated with phosphate, the equivalence point being detected from the change in emission intensity. Errors due to consumption of solution by the atomizer can be kept low by using short spraying times and low galvanometer damping. The average error is about -1% for 0.1M solutions and less than -5% for 0.01M. The method gives good results in the presence of titanium(III), zirconium, thorium and aluminium but cerium(III) and yttrium seriously interfere. PMID:18960392

  13. N-substituted phenothiazines as redox indicators in titrations with chloramine-T and chloramine-B.

    PubMed

    Gowda, H S; Mohan, B M; Ahmed, S A

    1980-12-01

    Profenamine hydrochloride, fluphenazine dihydrochloride, trifluopromazine hydrochloride, cyamepromazine maleate, perphenazine dihydrochloride and mepazine hydrochloride are proposed as redox indicators in the titration of hydroquinone, metol and ascorbic acid with chloramine-T and chloramine-B in sulphuric, hydrochloric and acetic acid media. They give a sharp reversible colour change at the equivalence point. A simple but accurate method for the determination of hydroquinone, metol and ascorbic acid is described. The conditional potentials and molar absorptivities of the indicators and redox potential of chloramine-B are reported.

  14. An elevated temperature titration calorimeter

    SciTech Connect

    Smith, J.R.; Zanonato, P.L.; Choppin, G.R. . Dept. of Chemistry)

    1991-06-01

    A variable-temperature (313 K to 353 K) titration calorimeter of high sensitivity has been constructed. The purpose of the calorimeter is to study temperature effects on the enthalpies of complex formation and of other reactions of metal cations such as hydrolysis and precipitation. Operation of the calorimetric system, including that final calculation of the heat released during titration, is automatic via computer control. Calibration tests of the calorimeter using 2-amino-2-hydroxymethyl-1,3-propanediol gave -(46.0 {plus minus} 0.3) kJ mol{sup {minus}1} and -(46.2 {plus minus} 0.2) kJ mol{sup {minus}1} for the enthalpy of protonation, at 318 K and at 343 K, respectively. For titrations of 2-bis(2-hydroxyethyl) amino-2-hydroxymethyl-1,3-propanediol, enthalpy of protonation values of -(28.4 {plus minus} 0.3) kJ mol{sup {minus}1} and -(29.3 {plus minus} 0.2) kJ mol{sup {minus}1} were obtained at 318 K and at 343 K, respectively. 6 refs., 3 figs., 2 tabs.

  15. A novel approach for high precision rapid potentiometric titrations: application to hydrazine assay.

    PubMed

    Sahoo, P; Malathi, N; Ananthanarayanan, R; Praveen, K; Murali, N

    2011-11-01

    We propose a high precision rapid personal computer (PC) based potentiometric titration technique using a specially designed mini-cell to carry out redox titrations for assay of chemicals in quality control laboratories attached to industrial, R&D, and nuclear establishments. Using this technique a few microlitre of sample (50-100 μl) in a total volume of ~2 ml solution can be titrated and the waste generated after titration is extremely low comparing to that obtained from the conventional titration technique. The entire titration including online data acquisition followed by immediate offline analysis of data to get information about concentration of unknown sample is completed within a couple of minutes (about 2 min). This facility has been created using a new class of sensors, viz., pulsating sensors developed in-house. The basic concept in designing such instrument and the salient features of the titration device are presented in this paper. The performance of the titration facility was examined by conducting some of the high resolution redox titrations using dilute solutions--hydrazine against KIO(3) in HCl medium, Fe(II) against Ce(IV) and uranium using Davies-Gray method. The precision of titrations using this innovative approach lies between 0.048% and 1.0% relative standard deviation in different redox titrations. With the evolution of this rapid PC based titrator it was possible to develop a simple but high precision potentiometric titration technique for quick determination of hydrazine in nuclear fuel dissolver solution in the context of reprocessing of spent nuclear fuel in fast breeder reactors.

  16. Biochemical titration of glycogen in vitro.

    PubMed

    Pelletier, Joffrey; Bellot, Grégory; Pouysségur, Jacques; Mazure, Nathalie M

    2013-01-01

    Glycogen is the main energetic polymer of glucose in vertebrate animals and plays a crucial role in whole body metabolism as well as in cellular metabolism. Many methods to detect glycogen already exist but only a few are quantitative. We describe here a method using the Abcam Glycogen assay kit, which is based on specific degradation of glycogen to glucose by glucoamylase. Glucose is then specifically oxidized to a product that reacts with the OxiRed probe to produce fluorescence. Titration is accurate, sensitive and can be achieved on cell extracts or tissue sections. However, in contrast to other techniques, it does not give information about the distribution of glycogen in the cell. As an example of this technique, we describe here the titration of glycogen in two cell lines, Chinese hamster lung fibroblast CCL39 and human colon carcinoma LS174, incubated in normoxia (21% O2) versus hypoxia (1% O2). We hypothesized that hypoxia is a signal that prepares cells to synthesize and store glycogen in order to survive(1).

  17. Biochemical Titration of Glycogen In vitro

    PubMed Central

    Pelletier, Joffrey; Bellot, Grégory; Pouysségur, Jacques; Mazure, Nathalie M.

    2013-01-01

    Glycogen is the main energetic polymer of glucose in vertebrate animals and plays a crucial role in whole body metabolism as well as in cellular metabolism. Many methods to detect glycogen already exist but only a few are quantitative. We describe here a method using the Abcam Glycogen assay kit, which is based on specific degradation of glycogen to glucose by glucoamylase. Glucose is then specifically oxidized to a product that reacts with the OxiRed probe to produce fluorescence. Titration is accurate, sensitive and can be achieved on cell extracts or tissue sections. However, in contrast to other techniques, it does not give information about the distribution of glycogen in the cell. As an example of this technique, we describe here the titration of glycogen in two cell lines, Chinese hamster lung fibroblast CCL39 and human colon carcinoma LS174, incubated in normoxia (21% O2) versus hypoxia (1% O2). We hypothesized that hypoxia is a signal that prepares cells to synthesize and store glycogen in order to survive1. PMID:24300406

  18. A Low-Cost Device for Automatic Photometric Titrations

    NASA Astrophysics Data System (ADS)

    Rocha, Fábio R. P.; Reis, Boaventura F.

    2000-02-01

    Electronics is an important topic in chemistry courses. However, the introduction of basic concepts is often difficult and the lab instruments are frequently seen as "black boxes". To address this problem, we propose the construction of a simple, low-cost (about $150 U.S.) automatic photometric titrator employing a light-emitting diode (LED) and a phototransistor. The electronic circuit can be assembled by the students themselves. The device was employed to implement a common procedure in chemical labs, making feasible the introduction of concepts related to electronics in undergraduate chemistry courses. The titrator is able to work automatically, since a feedback system permits stopping the addition of titrant solution when the end-point is achieved. With this demonstration, it can be stressed that automatic procedures can be implemented without expensive instruments. Additionally, a classical procedure becomes more attractive to the students and its importance to chemical analysis can be emphasized. The feasibility of the titrator was demonstrated by acid-base titrations of HCl solutions with NaOH in the presence of phenolphthalein and by iodimetric determination of ascorbic acid in vitamin C tablets and lemon juice. Precise results (0.7% relative standard deviation, n = 10) in agreement at the 95% confidence level with those attained by a conventional procedure were obtained.

  19. Analytical method for determination of benzene-arsenic acids

    SciTech Connect

    Mitchell, G.L.; Bayse, G.S.

    1988-01-01

    A sensitive analytical method has been modified for use in determination of several benzenearsonic acids, including arsanilic acid (p-aminobenzenearsonic acid), Roxarsone (3-nitro-4-hydroxybenzenearsonic acid), and p-ureidobenzene arsonic acid. Controlled acid hydrolysis of these compounds produces a quantitative yield of arsenate, which is measured colorimetrically as the molybdenum blue complex at 865 nm. The method obeys Beer's Law over the micromolar concentration range. These benzenearsonic acids are routinely used as feed additives in poultry and swine. This method should be useful in assessing tissue levels of the arsenicals in appropriate extracts.

  20. Method of increasing conversion of a fatty acid to its corresponding dicarboxylic acid

    DOEpatents

    Craft, David L.; Wilson, C. Ron; Eirich, Dudley; Zhang, Yeyan

    2004-09-14

    A nucleic acid sequence including a CYP promoter operably linked to nucleic acid encoding a heterologous protein is provided to increase transcription of the nucleic acid. Expression vectors and host cells containing the nucleic acid sequence are also provided. The methods and compositions described herein are especially useful in the production of polycarboxylic acids by yeast cells.

  1. Single-shot titrations and reaction monitoring by slice-selective NMR spectroscopy.

    PubMed

    Niklas, T; Stalke, D; John, M

    2015-01-25

    A new method, based on slice-selective NMR spectroscopy of inhomogeneous mixtures, is introduced to perform NMR titrations and reaction monitoring in a single experiment. The method was applied to the titration of a lithium salt with 12-crown-4, and to the reaction of nBuLi with N,N,N',N'',N''-pentamethyldiethylenetriamine (PMDTA).

  2. Molecular Titration Promotes Oscillations and Bistability in Minimal Network Models with Monomeric Regulators.

    PubMed

    Cuba Samaniego, Christian; Giordano, Giulia; Kim, Jongmin; Blanchini, Franco; Franco, Elisa

    2016-04-15

    Molecular titration is emerging as an important biochemical interaction mechanism within synthetic devices built with nucleic acids and the CRISPR/Cas system. We show that molecular titration in the context of feedback circuits is a suitable mechanism to enhance the emergence of oscillations and bistable behaviors. We consider biomolecular modules that can be inhibited or activated by input monomeric regulators; the regulators compete with constitutive titrating species to determine the activity of their target. By tuning the titration rate and the concentration of titrating species, it is possible to modulate the delay and convergence speed of the transient response, and the steepness and dead zone of the stationary response of the modules. These phenomena favor the occurrence of oscillations when modules are interconnected to create a negative feedback loop; bistability is favored in a positive feedback interconnection. Numerical simulations are supported by mathematical analysis showing that the capacity of the closed loop systems to exhibit oscillations or bistability is structural.

  3. Theoretical problems associated with the use of acetic anhydride as a co-solvent for the non-aqueous titration of hydrohalides of organic bases and quaternary ammonium salts.

    PubMed

    Völgyi, Gergely; Béni, Szabolcs; Takács-Novák, Krisztina; Görög, Sándor

    2010-01-01

    A potentiometric titration study of organic base hydrohalides and quaternary ammonium salts using perchloric acid as the titrant and a mixture of acetic anhydride and acetic acid as the solvent was carried out and the titration mixture was analysed by NMR in order to clarify the chemistry of the reactions involved. It was found that in contrast to the general belief the formation of acetyl halides and titratable free acetate ion does not take place prior to the titration but NMR spectra proved the formation of acetyl halides in the course of the titration. This observation and the fact that the shape of the titration curves depends on the nature of the hydrohaloic acid bound to the base or of the anion in the quaternary ammonium salts led to the conclusion that the titrating agent is acetyl perchlorate formed in situ during the titration. Equations of the reactions involved in the titration process are shown in the paper.

  4. 40 CFR Appendix A-4 to Part 60 - Test Methods 6 through 10B

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... Maintenance Requirements) of 40 CFR part 60, subpart A (General Provisions). Specific uses of these test... sulfur acid mist, are separated. The SO2 fraction is measured by the barium-thorin titration method. 3.0... precision of normality determination 11.2.3 Replicate titrations Ensure precision of...

  5. Anion-exchange nanospheres as titration reagents for anionic analytes.

    PubMed

    Zhai, Jingying; Xie, Xiaojiang; Bakker, Eric

    2015-08-18

    We present here anion-exchange nanospheres as novel titration reagents for anions. The nanospheres contain a lipophilic cation for which the counterion is initially Cl(-). Ion exchange takes place between Cl(-) in the nanospheres and a more lipophilic anion in the sample, such as ClO4(-) and NO3(-). Consecutive titration in the same sample solution for ClO4(-) and NO3(-) were demonstrated. As an application, the concentration of NO3(-) in spinach was successfully determined using this method.

  6. Risks and Benefits of Rapid Clozapine Titration.

    PubMed

    Lochhead, Jeannie D; Nelson, Michele A; Schneider, Alan L

    2016-05-18

    Clozapine is often considered the gold standard for the treatment of schizophrenia. Clinical guidelines suggest a gradual titration over 2 weeks to reduce the risks of adverse events such as seizures, hypotension, agranulocytosis, and myocarditis. The slow titration often delays time to therapeutic response. This raises the question of whether, in some patients, it may be safe to use a more rapid clozapine titration. The following case illustrates the potential risks associated with the use of multiple antipsychotics and rapid clozapine titration. We present the case of a young man with schizophrenia who developed life threatening neuroleptic malignant syndrome (NMS) during rapid clozapine titration and treatment with multiple antipsychotics. We were unable to find another case in the literature of NMS associated with rapid clozapine titration. This case is meant to urge clinicians to carefully evaluate the risks and benefits of rapid clozapine titration, and to encourage researchers to further evaluate the safety of rapid clozapine titration. Rapid clozapine titration has implications for decreasing health care costs associated with prolonged hospitalizations, and decreasing the emotional suffering associated with uncontrolled symptoms of psychosis. Clozapine is considered the most effective antipsychotic available thus efforts should focus on developing strategies that would allow for safest and most efficient use of clozapine to encourage its utilization for treatment resistance schizophrenia.

  7. Flow cytometric titration of retroviral expression vectors: comparison of methods for analysis of immunofluorescence histograms derived from cells expressing low antigen levels.

    PubMed

    Sladek, T L; Jacobberger, J W

    1993-01-01

    Few quantitative studies addressing immunofluorescence histogram analysis have been published. One study by Overton (Cytometry 9:619-626, 1988) has shown threshold and histogram subtraction methods to be accurate for analysis of well-separated immunofluorescence distributions of positive and negative cells. An evaluation of methods to analyze immunofluorescence histograms when positive and negative immunofluorescence distributions overlap has not, to our knowledge, been reported. In this paper, data obtained from flow cytometry of immunofluorescently stained cells infected with recombinant retroviruses that produce a range of simian virus 40 large T antigen levels were analyzed by threshold, histogram subtraction, and distribution modeling methods. This analysis showed that as the separation between the immunofluorescence distributions of positive and negative cell populations decrease the best methods for histogram analysis are modeling followed, in order, by histogram subtraction, and threshold analysis.

  8. [Simultaneous determination of calcium and magnesium by calculative spectrophotometric complexometric titration].

    PubMed

    Liao, Li-fu; Xiao, Xi-lin; Yang, Ming-hui; Yang, Jing

    2007-12-01

    A new spectrophotometric complexometric titration method coupled with chemometrics for the determination of mixtures of metal ions has been developed. In the method described here, the titrant is a mixture of EDTA and two indicators. In the process of titration, both the volumetric addition of titrant and the progress of titration reaction can be characterized simultaneously by chemometric calculation with the absorption spectra, and then the titration curves can be obtained. With the titration curves, a matrix equation can be established, and thus the concentration of each component in the mixture of metal ions can be calculated with principal component regression. The method only needs the information of absorption spectra to obtain the analytical results, and is free of volumetric measurements. So the method is simple, convenient and precise, and has been applied to the simultaneous determination of mixtures of calcium and magnesium using malachite green and Cu-PAN as indicators with satisfactory results.

  9. Chemical force titrations of functionalized Si(111) surfaces.

    PubMed

    Mengistu, Tadesse Z; Goel, Vishya; Horton, J Hugh; Morin, Sylvie

    2006-06-01

    Chemical force titrations-plots of the adhesive force between an atomic force microscope tip and sample as a function of pH-were acquired on alkyl monolayer-derivatized Si(111) surfaces. Gold-coated AFM tips modified with thioalkanoic acid self-assembled monolayers (SAM) were employed. Alkyl monolayer-derivatized Si(111) surfaces terminated with methyl, carboxyl, and amine groups were produced via hydrosilylation reactions between 1-alkene reagents and H-terminated silicon. The functionalized surfaces were characterized using standard surface science techniques (AFM, FTIR, and XPS). Titration of the methyl-terminated surface using the modified (carboxyl-terminated) atomic force microscope tip resulted in a small pH-independent hydrophobic interaction. Titration of the amine-terminated surface using the same tip resulted in the determination of a surface pKa of 5.8 for the amine from the pH value from the maximum in the force titration curve. A pK(1/2) of 4.3 was determined for the carboxyl-terminated Si(111) in a similar way. These results will be discussed in relation to the modified Si(111) surface chemistry and organic layer structure, as well as with respect to existing results on Au surfaces modified with SAMs bearing the same functional groups.

  10. Isothermal Titration Calorimetry Can Provide Critical Thinking Opportunities

    ERIC Educational Resources Information Center

    Moore, Dale E.; Goode, David R.; Seney, Caryn S.; Boatwright, Jennifer M.

    2016-01-01

    College chemistry faculties might not have considered including isothermal titration calorimetry (ITC) in their majors' curriculum because experimental data from this instrumental method are often analyzed via automation (software). However, the software-based data analysis can be replaced with a spreadsheet-based analysis that is readily…

  11. Quantification of reverse transcriptase activity by real-time PCR as a fast and accurate method for titration of HIV, lenti- and retroviral vectors.

    PubMed

    Vermeire, Jolien; Naessens, Evelien; Vanderstraeten, Hanne; Landi, Alessia; Iannucci, Veronica; Van Nuffel, Anouk; Taghon, Tom; Pizzato, Massimo; Verhasselt, Bruno

    2012-01-01

    Quantification of retroviruses in cell culture supernatants and other biological preparations is required in a diverse spectrum of laboratories and applications. Methods based on antigen detection, such as p24 for HIV, or on genome detection are virus specific and sometimes suffer from a limited dynamic range of detection. In contrast, measurement of reverse transcriptase (RT) activity is a generic method which can be adapted for higher sensitivity using real-time PCR quantification (qPCR-based product-enhanced RT (PERT) assay). We present an evaluation of a modified SYBR Green I-based PERT assay (SG-PERT), using commercially available reagents such as MS2 RNA and ready-to-use qPCR mixes. This assay has a dynamic range of 7 logs, a sensitivity of 10 nU HIV-1 RT and outperforms p24 ELISA for HIV titer determination by lower inter-run variation, lower cost and higher linear range. The SG-PERT values correlate with transducing and infectious units in HIV-based viral vector and replication-competent HIV-1 preparations respectively. This assay can furthermore quantify Moloney Murine Leukemia Virus-derived vectors and can be performed on different instruments, such as Roche Lightcycler® 480 and Applied Biosystems ABI 7300. We consider this test to be an accurate, fast and relatively cheap method for retroviral quantification that is easily implemented for use in routine and research laboratories.

  12. Quantification of Reverse Transcriptase Activity by Real-Time PCR as a Fast and Accurate Method for Titration of HIV, Lenti- and Retroviral Vectors

    PubMed Central

    Vermeire, Jolien; Naessens, Evelien; Vanderstraeten, Hanne; Landi, Alessia; Iannucci, Veronica; Van Nuffel, Anouk; Taghon, Tom; Pizzato, Massimo; Verhasselt, Bruno

    2012-01-01

    Quantification of retroviruses in cell culture supernatants and other biological preparations is required in a diverse spectrum of laboratories and applications. Methods based on antigen detection, such as p24 for HIV, or on genome detection are virus specific and sometimes suffer from a limited dynamic range of detection. In contrast, measurement of reverse transcriptase (RT) activity is a generic method which can be adapted for higher sensitivity using real-time PCR quantification (qPCR-based product-enhanced RT (PERT) assay). We present an evaluation of a modified SYBR Green I-based PERT assay (SG-PERT), using commercially available reagents such as MS2 RNA and ready-to-use qPCR mixes. This assay has a dynamic range of 7 logs, a sensitivity of 10 nU HIV-1 RT and outperforms p24 ELISA for HIV titer determination by lower inter-run variation, lower cost and higher linear range. The SG-PERT values correlate with transducing and infectious units in HIV-based viral vector and replication-competent HIV-1 preparations respectively. This assay can furthermore quantify Moloney Murine Leukemia Virus-derived vectors and can be performed on different instruments, such as Roche Lightcycler® 480 and Applied Biosystems ABI 7300. We consider this test to be an accurate, fast and relatively cheap method for retroviral quantification that is easily implemented for use in routine and research laboratories. PMID:23227216

  13. Behaviour of ferroin indicator in iron(ii)- dichromate titrations A Critical study.

    PubMed

    Sriramam, K

    1972-10-01

    A critical evaluation has been made of the optimum conditions for a successful direct and reverse titration of iron(II) with dichromate, with ferroin as indicator, taking into account the influence of acidity, the catalytic effect of iron(III) and the induction effect of the iron(II)-dichromate reaction on the indicator oxidation reaction. The difference in the behaviour of the indicator in the direct and reverse titrations is believed to be due to the presence of a slow : step in the indicator oxidation mechanism ; this is however surmounted in the reverse titration, making it possible to use the indicator even in a 2N sulphuric add medium for this titration. The higher potentials recorded in the reverse titration are believed to be due to the oxide film on the platinum metal electrode. While iron(III) seems to help lower the kinetic barrier of the slow step sufficiently in the indicator oxidation mechanism, the ferriin-ferroin system seems to help m a similar way toe establishment of a more positive potential of the Ct(VI)/Cr(III) system at the platinum electrode in me direct titration; this value explains the feasibility of the direct visual titration in 2N sulphuric acid, although the colour change at the end-point is slow for other reasons.

  14. Isothermal Titration Calorimetry to Characterize Enzymatic Reactions.

    PubMed

    Mazzei, Luca; Ciurli, Stefano; Zambelli, Barbara

    2016-01-01

    Isothermal titration calorimetry (ITC) is a technique that measures the heat released or absorbed during a chemical reaction as an intrinsic probe to characterize any chemical process that involves heat changes spontaneously occurring during the reaction. The general features of this method to determine the kinetic and thermodynamic parameters of enzymatic reactions (kcat, KM, ΔH) are described and discussed here together with some detailed applications to specific cases. ITC does not require any modification or labeling of the system under analysis, can be performed in solution, and needs only small amounts of enzyme. These properties make ITC an invaluable, powerful, and unique tool to extend the knowledge of enzyme kinetics to drug discovery.

  15. A Tabular Approach to Titration Calculations

    ERIC Educational Resources Information Center

    Lim, Kieran F.

    2012-01-01

    Titrations are common laboratory exercises in high school and university chemistry courses, because they are easy, relatively inexpensive, and they illustrate a number of fundamental chemical principles. While students have little difficulty with calculations involving a single titration step, there is a significant leap in conceptual difficulty…

  16. Advances in nucleic acid-based detection methods.

    PubMed Central

    Wolcott, M J

    1992-01-01

    Laboratory techniques based on nucleic acid methods have increased in popularity over the last decade with clinical microbiologists and other laboratory scientists who are concerned with the diagnosis of infectious agents. This increase in popularity is a result primarily of advances made in nucleic acid amplification and detection techniques. Polymerase chain reaction, the original nucleic acid amplification technique, changed the way many people viewed and used nucleic acid techniques in clinical settings. After the potential of polymerase chain reaction became apparent, other methods of nucleic acid amplification and detection were developed. These alternative nucleic acid amplification methods may become serious contenders for application to routine laboratory analyses. This review presents some background information on nucleic acid analyses that might be used in clinical and anatomical laboratories and describes some recent advances in the amplification and detection of nucleic acids. PMID:1423216

  17. Comparison of RNA hybridization, hemagglutination assay, titration of infectious virus and immunofluorescence as methods for monitoring influenza virus replication in vitro.

    PubMed

    Rimmelzwaan, G F; Baars, M; Claas, E C; Osterhaus, A D

    1998-09-01

    Rapid and sensitive methods for the monitoring of influenza virus replication in vitro are needed to address several research questions. Four methods based on different principles were compared: the hemagglutination (HA) assay, the measurement of virus infectivity titers in culture supernatants, the enumeration of infected cells by immunofluorescence and RNA hybridization techniques using digoxigenin (DIG) labeled RNA probes. To this end, MDCK cells were infected at different multiplicities of infection (moi) with a recent influenza A virus (A/Netherlands/18/94 H3N2) and the kinetics of virus replication were monitored with these four assays. At high moi, virus released into the culture supernatant of infected cells was detected by the HA assay 12 h post infection, whereas at lower moi (< or = 0.01) the first HA activity was not detected before 24 h post infection. The measurement of infectious viruses in the culture supernatant proved to be more sensitive, since 4-12 h post infection newly produced virus was detected depending on the moi used. This finding was in agreement with results obtained by the immunofluorescence assay using an antibody preparation specific for the nucleoprotein: single infected cells could be detected as early as 4 h post infection. At this time point, positive signals were also obtained when mRNA/cRNA specific hybridization was carried out for the NP gene segment, but not for viral NP RNA or RNA specific for the hemagglutinin, which were only detected at later time points after infection. Thus, besides direct measurement of infectious virus and immunofluorescence, RNA hybridization proved to be a sensitive assay for monitoring influenza virus replication in vitro.

  18. Development of highly sensitive cadmium iondashselective electrodes by titration method and its application to cadmium ion determination in industrial waste water.

    PubMed

    Ito, S; Asano, Y; Wada, H

    1997-04-01

    Characteristics of cadmium iondashselective electrode made cadmium sulphide (CdS)-silver sulphide (Ag(2)S) mixture were studied. CdS-Ag(2)S mixtures were obtained by gas/solid-phase reaction between silver-cadmium mixed powder and hydrogen sulphide gas (dry method) and by ionic reaction between cadmium-silver mixed ions and sulphide ion (wet method). As a result, it was found that the CdS-Ag(2)S mixture had to be made in the condition of excess existence of sulfur and had better regulate the excess sulfur quantity minimum, for the CdS-Ag(2)S pressed membrane gave a good Nernstian response against the cadmium ion concentration change. As the best way, CdS-Ag(2)S mixture was obtained by adding sulphide ion solution to 5 mol% cadmium ion and 95 mol% silver ion mixed solution while measuring silver sulphide (Ag(2)S) electrode potential as an indicator electrode. According to the reaction was stopped when the potential variation from the initial potential in the sulphide ion solution reached at 87-116 mV which the sulphide ion concentration became 10(-3) - 10(-4) of the initial concentration, the cadmium ion membrane pressed diameter of 8 mm and thickness of 2 mm showed a Nernstian response from 10(-8) to 10(-1) M of cadmium ion concentration. Furthermore, aiming to its application for industrial waste water, masking buffer for interfering metal ions such as lead ion (Pb(2+)) and copper ion (Cu(2+)), which were possibly coexisted and to adjust total ionic strength and pH of sample was developed. The present Cd(2+) iondashselective electrode was applied to the determination of Cd(2+) in the industrial waste water. The good regression line with correlation factor of 0.984 was obtained compared with the conventional atomic absorption spectroscopy. PMID:18966791

  19. Indirect method for prediction of hemagglutination inhibition antibody titers to Newcastle disease virus in chickens by titration of antibodies in egg yolk.

    PubMed

    Yeo, Sang-Geon; Nagy, Eva; Krell, Peter J

    2003-03-01

    Attempts were made to establish methods for indirect prediction of hemagglutination inhibition (HI) antibody titers to Newcastle disease virus (NDV) in sera of laying hens and day-old chicks by determining if these are correlated to HI titers in egg yolks. For this purpose, geometric means of HI antibody titers in sera from 60 hens, yolks from 60 matched eggs, and sera from 180 day-old chicks of an identical vaccination program were measured and plotted. There was a significant correlation between HI antibody titers in yolks (X) and hens (Y), with a linear regression of Y = 23.24 + 0.47X and a correlation coefficient of r = 0.65. The linear regression between HI antibody titers in yolks (X) and chicks (Y) was Y = 6.33 + 0.36X (r = 0.58). Immunity to NDV in hens and their offspring can be maintained effectively, and the proper time for the vaccination or booster can be determined by reference to HI titers predicted from the linear regression in the present study. The approach of testing egg yolk for HI titers provides a feasible alternative to determining HI titers from blood samples and eliminates stress in birds during blood sampling.

  20. Using RNA sample titrations to assess microarray platform performance and normalization techniques.

    PubMed

    Shippy, Richard; Fulmer-Smentek, Stephanie; Jensen, Roderick V; Jones, Wendell D; Wolber, Paul K; Johnson, Charles D; Pine, P Scott; Boysen, Cecilie; Guo, Xu; Chudin, Eugene; Sun, Yongming Andrew; Willey, James C; Thierry-Mieg, Jean; Thierry-Mieg, Danielle; Setterquist, Robert A; Wilson, Mike; Lucas, Anne Bergstrom; Novoradovskaya, Natalia; Papallo, Adam; Turpaz, Yaron; Baker, Shawn C; Warrington, Janet A; Shi, Leming; Herman, Damir

    2006-09-01

    We have assessed the utility of RNA titration samples for evaluating microarray platform performance and the impact of different normalization methods on the results obtained. As part of the MicroArray Quality Control project, we investigated the performance of five commercial microarray platforms using two independent RNA samples and two titration mixtures of these samples. Focusing on 12,091 genes common across all platforms, we determined the ability of each platform to detect the correct titration response across the samples. Global deviations from the response predicted by the titration ratios were observed. These differences could be explained by variations in relative amounts of messenger RNA as a fraction of total RNA between the two independent samples. Overall, both the qualitative and quantitative correspondence across platforms was high. In summary, titration samples may be regarded as a valuable tool, not only for assessing microarray platform performance and different analysis methods, but also for determining some underlying biological features of the samples.

  1. Potentiometric titrations in acetonitrile-water mixtures: evaluation of aqueous ionisation constant of ketoprofen.

    PubMed

    Herrador, M Angeles; González, A Gustavo

    2002-03-11

    Non ideality of acetonitrile-water mixtures was studied from data on the excess of molar volumes and viscosities. pH and autoprotolisis constants were evaluated at the standard state of the mixed solvent from titrations of a strong acid with a strong base. In order to illustrate the evaluation of the aqueous ionisation constant of water insoluble compounds from pH titrations in ACN-water mixtures, a typical insoluble arylpropionic acid, ketoprofen, was chosen. Ketoprofen was titrated in mixtures from 10 to 70% w/w of acetonitrile against a strong base. From the titration data, the ionisation constant of ketoprofen was evaluated at the standard state of the solvent mixture (pK(a)(*)). Aqueous pK(a) was determined by extrapolation, as the intercept of the plot of pK(a)(*) versus ACN mole fraction.

  2. Kinetics of bacterial potentiometric titrations: the effect of equilibration time on buffering capacity of Pantoea agglomerans suspensions.

    PubMed

    Kapetas, Leon; Ngwenya, Bryne T; Macdonald, Alan M; Elphick, Stephen C

    2011-07-15

    Several recent studies have made use of continuous acid-base titration data to describe the surface chemistry of bacterial cells as a basis for accurately modelling metal adsorption to bacteria and other biomaterials of potential industrial importance. These studies do not share a common protocol; rather they titrate in different pH ranges and they use different stability criteria to define equilibration time during titration. In the present study we investigate the kinetics of bacterial titrations and test the effect they have on the derivation of functional group concentrations and acidity constants. We titrated suspensions of Pantoea agglomerans by varying the equilibration time between successive titrant additions until stability of 0.1 or 0.001 mV s(-1) was attained. We show that under longer equilibration times, titration results are less reproducible and suspensions exhibit marginally higher buffering. Fluorescence images suggest that cell lysis is not responsible for these effects. Rather, high DOC values and titration reversibility hysterisis after long equilibration times suggest that variability in buffering is due to the presence of bacterial exudates, as demonstrated by titrating supernatants separated from suspensions of different equilibration times. It is recommended that an optimal equilibration time is always determined with variable stability control and preliminary reversibility titration experiments.

  3. Method for nucleic acid isolation using supercritical fluids

    DOEpatents

    Nivens, David E.; Applegate, Bruce M.

    1999-01-01

    A method for detecting the presence of a microorganism in an environmental sample involves contacting the sample with a supercritical fluid to isolate nucleic acid from the microorganism, then detecting the presence of a particular sequence within the isolated nucleic acid. The nucleic acid may optionally be subjected to further purification.

  4. Method for nucleic acid isolation using supercritical fluids

    DOEpatents

    Nivens, D.E.; Applegate, B.M.

    1999-07-13

    A method is disclosed for detecting the presence of a microorganism in an environmental sample involves contacting the sample with a supercritical fluid to isolate nucleic acid from the microorganism, then detecting the presence of a particular sequence within the isolated nucleic acid. The nucleic acid may optionally be subjected to further purification. 4 figs.

  5. Limitations of the potentiometric titration technique in determining the proton active site density of goethite surfaces

    NASA Astrophysics Data System (ADS)

    Lützenkirchen, Johannes; Boily, Jean-François; Lövgren, Lars; Sjöberg, Staffan

    2002-10-01

    Density of proton active surface sites at mineral surfaces is a property of fundamental importance in equilibrium modeling of surface complexation reactions. In this article, methods for an experimental determination of these sites at the surface of α-FeOOH (goethite) are explored. It is shown that previously obtained saturation data of goethite with respect to protons do not yield a site density that can be considered as an intrinsic sorbent property: the results are below crystallographically expected values and values for different ionic media in terms of composition and concentration yield different numbers - for example, chloride would yield higher values than nitrate at the same concentration, and higher electrolyte concentration would favor higher apparent maxima. Although site saturation might be explained by electrostatic repulsion, which is more efficient at high electrolyte concentration or for certain ions, further independent experimental results show that no saturation occurs on goethite down to ph ≡ -log[H +] = 2.2 and possibly to ph = 1.0 in 0.6 M NaCl. For those very low pH values, the experimental charging curve was obtained by coulometric back titration (using the Gran plot) or titrations with tris (hydroxymethyl)-aminomethane of the supernatant of acidified goethite suspension. These experimental data are to our knowledge the first high quality data at such low pHs. However, small errors in the determination of proton concentrations (1%) are shown to strongly affect the shape of the charging curve for ph < 2. Furthermore, goethite dissolution (proton consumption and iron reduction in coulometric titrations) and liquid junction effects interfere at low ph, hampering the straightforward application of coulometric Gran titrations over the whole pH range. From these experiments, it can nonetheless be ascertained that a minimum of 2.5 protons/nm 2 can be adsorbed at the goethite surface from the point of zero charge (ph 9.4) to pH 0.9. Although

  6. Spectrophotometric method for determining gibberellic acid in fermentation broths.

    PubMed

    Berríos, Julio; Illanes, Andrés; Aroca, Germán

    2004-01-01

    A novel method for the quantitative determination of gibberellic acid in fermentation broths has been developed. It is based on the kinetic of the reaction of conversion of gibberellic acid to gibberellenic acid. The method is simple, reliable, faster than most of methods known, and free of the interferences which commonly affect spectrophotometric methods currently in use. Its threshold sensitivity is 0.1 g and its accuracy is greater than 97% for concentrations of gibberellic acid ranging from 0.1 to 1 g l(-1).

  7. Comparison of Rapid Methods for Analysis of Bacterial Fatty Acids

    PubMed Central

    Moss, C. Wayne; Lambert, M. A.; Merwin, W. H.

    1974-01-01

    When rapid gas-liquid chromatography methods for determination of bacterial fatty acids were compared, results showed that saponification was required for total fatty acid analysis. Transesterification with boron-trihalide reagents (BF3-CH3OH, BCl3-CH3OH) caused extensive degradation of cyclopropane acids and was less effective than saponification in releasing cellular hydroxy fatty acids. Digestion of cells with tetramethylammonium hydroxide was unsatisfactory because of extraneous gas-liquid chromatography peaks and because of lower recovery of branched-chain and hydroxy fatty acids. A simple, rapid saponification procedure which can be used for total cellular fatty acid analysis of freshly grown cells is described. PMID:4844271

  8. The Softening of Hard Water and Complexometric Titrations. An Undergraduate Experiment

    NASA Astrophysics Data System (ADS)

    Ceretti, Helena; Hughes, Enrique A.; Zalts, Anita

    1999-10-01

    A 2-hour experiment for undergraduates is presented in which (i) water hardness is explained and demonstrated; (ii) ion-exchange resin properties are visually demonstrated and then used for softening water; (iii) complexometric titrations are used for evaluating water hardness before and after softening; and (iv) acid-base titration can be used to show that the Ca/Mg ions removed by the ion exchanger are replaced by H ions.

  9. Redox potentials of tungsten and its alloying partners-fundamentals of tungsten determination by redox titration.

    PubMed

    Wünsch, G

    1980-08-01

    The fundamentals of tungsten determination by redox titration in hydrochloric acid medium are describe. Conditional redox potentials are given for W, Cr, V, Fe, Ti, Mo, Cu and Sn as a function of HCl concentration. The basic experimental conditions and the difference in conditional potentials required for consecutive titrations are discussed. Any mixture of tungsten with other metals can be analysed without separation if it does not contain both Cr and Ti or both Mo and V.

  10. Method for identifying and quantifying nucleic acid sequence aberrations

    DOEpatents

    Lucas, J.N.; Straume, T.; Bogen, K.T.

    1998-07-21

    A method is disclosed for detecting nucleic acid sequence aberrations by detecting nucleic acid sequences having both a first and a second nucleic acid sequence type, the presence of the first and second sequence type on the same nucleic acid sequence indicating the presence of a nucleic acid sequence aberration. The method uses a first hybridization probe which includes a nucleic acid sequence that is complementary to a first sequence type and a first complexing agent capable of attaching to a second complexing agent and a second hybridization probe which includes a nucleic acid sequence that selectively hybridizes to the second nucleic acid sequence type over the first sequence type and includes a detectable marker for detecting the second hybridization probe. 11 figs.

  11. Method for identifying and quantifying nucleic acid sequence aberrations

    DOEpatents

    Lucas, Joe N.; Straume, Tore; Bogen, Kenneth T.

    1998-01-01

    A method for detecting nucleic acid sequence aberrations by detecting nucleic acid sequences having both a first and a second nucleic acid sequence type, the presence of the first and second sequence type on the same nucleic acid sequence indicating the presence of a nucleic acid sequence aberration. The method uses a first hybridization probe which includes a nucleic acid sequence that is complementary to a first sequence type and a first complexing agent capable of attaching to a second complexing agent and a second hybridization probe which includes a nucleic acid sequence that selectively hybridizes to the second nucleic acid sequence type over the first sequence type and includes a detectable marker for detecting the second hybridization probe.

  12. Continuous-flow free acid monitoring method and system

    DOEpatents

    Strain, J.E.; Ross, H.H.

    1980-01-11

    A free acid monitoring method and apparatus is provided for continuously measuring the excess acid present in a process stream. The disclosed monitoring system and method is based on the relationship of the partial pressure ratio of water and acid in equilibrium with an acid solution at constant temperature. A portion of the process stream is pumped into and flows through the monitor under the influence of gravity and back to the process stream. A continuous flowing sample is vaporized at a constant temperature and the vapor is subsequently condensed. Conductivity measurements of the condensate produces a nonlinear response function from which the free acid molarity of the sample process stream is determined.

  13. Continuous-flow free acid monitoring method and system

    DOEpatents

    Strain, James E.; Ross, Harley H.

    1981-01-01

    A free acid monitoring method and apparatus is provided for continuously measuring the excess acid present in a process stream. The disclosed monitoring system and method is based on the relationship of the partial pressure ratio of water and acid in equilibrium with an acid solution at constant temperature. A portion of the process stream is pumped into and flows through the monitor under the influence of gravity and back to the process stream. A continuous flowing sample is vaporized at a constant temperature and the vapor is subsequently condensed. Conductivity measurements of the condensate produces a nonlinear response function from which the free acid molarity of the sample process stream is determined.

  14. High throughput continuous titration based on a flow ratiometry controlled with feedback-based variable triangular waves and subsequent fixed triangular waves.

    PubMed

    Tanaka, Hideji; Baba, Tsutomu

    2005-10-15

    We propose a new approach for high-throughput continuous titration based on a flow ratiometry. The method was applied to potentiometric titrations of acids and bases. A base solution, the flow rate F(B) of which was varied in response to controller output voltage V(c), was merged with an acid solution under constant total (acid+base) flow rate. Downstream, the pH of the mixed solution was measured with a flow-through glass electrode. Initially, V(c), and thus F(B) was increased linearly. At the instant the equivalence point (EP) was sensed, the ramp direction of V(c) was reversed from upward to downward. The direction was reversed to upward when EP was sensed again. Such the feedback-based operation gives a triangular waveform of V(c), because there is a delay corresponding mainly to the transit time of merged solutions to reach the sensor. The value of V(c) that gives EP composition, V(E), was estimated by averaging the most recent maximum and minimum values of V(c). Next, fixed triangular waves of V(c) was used to control F(B). The amplitude and the scan rate of the waves were fixed narrower and faster, respectively, than those in the feedback-based operation in order to improve the throughput rate. The EP can be located as long as the scan range covers V(E). These automated processes limited the titration to just the narrow range around EP, and thus realized extremely high throughput rate of maximally 17.6 titrations per minute (=3.4s per titration) at R.S.D.=0.35%. PMID:18970249

  15. Titration of vaccinia virus by intravenous injection of chick embryos.

    PubMed

    KAPLAN, C

    1960-01-01

    The final test of a smallpox vaccine is its capacity to prevent the disease from developing in inoculated individuals. This capacity, however, cannot be measured directly, so that other methods of assessing the efficacy of vaccine have had to be developed. A laboratory method-pock counting on the chorio-allantoic membrane of chick embryos-has recently been shown to provide a reasonably reliable estimate of the number of infective units in a given vaccine. In this paper, the author compares this pock-counting method with another method-titration by intravenous injection of chick embryos. He concludes that, although the reproducibility of titrations by intravenous injection compares very favourably with that obtained by chorio-allantoic inoculation, the former method would not be advantageous for the assay of vaccines, since it is very time-consuming and since differences in virulence might obscure comparisons between the efficacy of vaccines.

  16. Applications of isothermal titration calorimetry in protein science.

    PubMed

    Liang, Yi

    2008-07-01

    During the past decade, isothermal titration calorimetry (ITC) has developed from a specialist method for understanding molecular interactions and other biological processes within cells to a more robust, widely used method. Nowadays, ITC is used to investigate all types of protein interactions, including protein-protein interactions, protein-DNA/RNA interactions, protein-small molecule interactions and enzyme kinetics; it provides a direct route to the complete thermodynamic characterization of protein interactions. This review concentrates on the new applications of ITC in protein folding and misfolding, its traditional application in protein interactions, and an overview of what can be achieved in the field of protein science using this method and what developments are likely to occur in the near future. Also, this review discusses some new developments of ITC method in protein science, such as the reverse titration of ITC and the displacement method of ITC.

  17. Sulfuric acid thermoelectrochemical system and method

    DOEpatents

    Ludwig, Frank A.

    1989-01-01

    A thermoelectrochemical system in which an electrical current is generated between a cathode immersed in a concentrated sulfuric acid solution and an anode immersed in an aqueous buffer solution of sodium bisulfate and sodium sulfate. Reactants consumed at the electrodes during the electrochemical reaction are thermochemically regenerated and recycled to the electrodes to provide continuous operation of the system.

  18. Integration and global analysis of isothermal titration calorimetry data for studying macromolecular interactions.

    PubMed

    Brautigam, Chad A; Zhao, Huaying; Vargas, Carolyn; Keller, Sandro; Schuck, Peter

    2016-05-01

    Isothermal titration calorimetry (ITC) is a powerful and widely used method to measure the energetics of macromolecular interactions by recording a thermogram of differential heating power during a titration. However, traditional ITC analysis is limited by stochastic thermogram noise and by the limited information content of a single titration experiment. Here we present a protocol for bias-free thermogram integration based on automated shape analysis of the injection peaks, followed by combination of isotherms from different calorimetric titration experiments into a global analysis, statistical analysis of binding parameters and graphical presentation of the results. This is performed using the integrated public-domain software packages NITPIC, SEDPHAT and GUSSI. The recently developed low-noise thermogram integration approach and global analysis allow for more precise parameter estimates and more reliable quantification of multisite and multicomponent cooperative and competitive interactions. Titration experiments typically take 1-2.5 h each, and global analysis usually takes 10-20 min.

  19. Characterisation of iron binding ligands in seawater by reverse titration.

    PubMed

    Hawkes, Jeffrey A; Gledhill, Martha; Connelly, Douglas P; Achterberg, Eric P

    2013-03-01

    Here we demonstrate the use of reverse titration - competitive ligand exchange-adsorptive cathodic stripping voltammetry (RT-CLE-ACSV) for the analysis of iron (Fe) binding ligands in seawater. In contrast to the forward titration, which examines excess ligands in solution, RT-CLE-ACSV examines the existing Fe-ligand complexes by increasing the concentration of added (electroactive) ligand (1-nitroso-2-naphthol) and analysis of the proportion of Fe bound to the added ligand. The data manipulation allows the accurate characterisation of ligands at equal or lower concentrations than Fe in seawater, and disregards electrochemically inert dissolved Fe such as some colloidal phases. The method is thus superior to the forward titration in environments with high Fe and low ligand concentrations or high concentrations of inert Fe. We validated the technique using the siderophore ligand ferrioxamine B, and observed a stability constant [Formula: see text] of 0.74-4.37×10(21) mol(-1), in agreement with previous results. We also successfully analysed samples from coastal waters and a deep ocean hydrothermal plume. Samples from these environments could not be analysed with confidence using the forward titration, highlighting the effectiveness of the RT-CLE-ACSV technique in waters with high concentrations of inert Fe.

  20. Asphalt compatibility testing using the automated Heithaus titration test

    SciTech Connect

    Pauli, A.T.

    1996-12-31

    The Heithaus titration test or variations of the test have been used for over 35 years to predict compatibilities of blends of asphalts from different crude sources. Asphalt compatibility is determined from three calculated parameters that measure the state of peptization of an asphalt or asphalt blend. The parameter p{sub a} is a measure of the peptizability of the asphaltenes. The parameter p{sub a} is a measure of the peptizing power of the maltenes, and the parameter P, derived from p{sub a} and p{sub o} values, is a measure of the overall state of peptization of the asphalt or asphalt blend. In Heithaus original procedure, samples of asphalt were dissolved in toluene and titrated with n-heptane in order to initiate flocculation. The onset of flocculation was detected either by photography or by spotting a filter paper with a small amount of the titrated solution. Recently, an {open_quotes}automated{close_quotes} procedure, after Hotier and Robin, has been developed for use with asphalt. In the automated method UV-visible spectrophotometric detection measures the onset of flocculation as a peak with the percent transmittance plotted as a function of the volume of titrating solvent added to a solution of asphalt. The automated procedure has proven to be less operator dependent and much faster than the original Heithaus procedure. Results from the automated procedure show the data to be consistent with results from the original, {open_quotes}classical{close_quotes} Heithaus procedure.

  1. Methods of refining and producing isomerized fatty acid esters and fatty acids from natural oil feedstocks

    DOEpatents

    Snead, Thomas E.; Cohen, Steven A.; Gildon, Demond L.; Beltran, Leslie V.; Kunz, Linda A.; Pals, Tessa M.; Quinn, Jordan R; Behrends, Jr., Raymond T.; Bernhardt, Randal J.

    2016-07-05

    Methods are provided for refining natural oil feedstocks and producing isomerized esters and acids. The methods comprise providing a C4-C18 unsaturated fatty ester or acid, and isomerizing the fatty acid ester or acid in the presence of heat or an isomerization catalyst to form an isomerized fatty ester or acid. In some embodiments, the methods comprise forming a dibasic ester or dibasic acid prior to the isomerizing step. In certain embodiments, the methods further comprise hydrolyzing the dibasic ester to form a dibasic acid. In certain embodiments, the olefin is formed by reacting the feedstock in the presence of a metathesis catalyst under conditions sufficient to form a metathesized product comprising olefins and esters, separating the olefins from the esters in the metathesized product, and transesterifying the esters in the presence of an alcohol to form a transesterified product having unsaturated esters.

  2. Titration of gold nanoparticles in phase extraction.

    PubMed

    Cheng, Han-Wen; Schadt, Mark J; Zhong, Chuan-Jian

    2015-12-01

    In the organic-aqueous phase transfer process of gold nanoparticles, there are two types of distinctive interfaces involving hydrophilic and hydrophobic ligands, the understanding of which is important for the design of functional nanomaterials for analytical/bioanalytical applications and the control over the nanoparticles' nanoactivity and nanotoxicity in different phases. This report describes new findings of an investigation of the quantitative aspect of ligand ion pairing at the capping monolayer structure that drives the phase extraction of gold nanoparticles. Alkanethiolate-capped gold nanoparticles of 8 nm diameter with high size monodispersity (RSD ∼ 5%) were first derivatized by a ligand place exchange reaction with 11-mercaptoundecanoic acid to form a mixed monolayer shell consisting of both hydrophobic (-CH3) and hydrophilic (-COOH) groups. It was followed by quantitative titration of the resulting nanoparticles with a cationic species (-NR4(+)) in a toluene phase, yielding ion pairing of -NR4(+) and -COO(-) on part of the capping monolayer. Analysis of the phase extraction allowed a quantitative determination of the percentage of ion pairing and structural changes in the capping monolayer on the nanoparticles. The results, along with morphological characterization, are discussed in terms of the interfacial structural changes and their implications on the rational design of surface-functionalized nanoparticles and fine tuning of the interfacial reactivity. PMID:26523548

  3. Titration of gold nanoparticles in phase extraction.

    PubMed

    Cheng, Han-Wen; Schadt, Mark J; Zhong, Chuan-Jian

    2015-12-01

    In the organic-aqueous phase transfer process of gold nanoparticles, there are two types of distinctive interfaces involving hydrophilic and hydrophobic ligands, the understanding of which is important for the design of functional nanomaterials for analytical/bioanalytical applications and the control over the nanoparticles' nanoactivity and nanotoxicity in different phases. This report describes new findings of an investigation of the quantitative aspect of ligand ion pairing at the capping monolayer structure that drives the phase extraction of gold nanoparticles. Alkanethiolate-capped gold nanoparticles of 8 nm diameter with high size monodispersity (RSD ∼ 5%) were first derivatized by a ligand place exchange reaction with 11-mercaptoundecanoic acid to form a mixed monolayer shell consisting of both hydrophobic (-CH3) and hydrophilic (-COOH) groups. It was followed by quantitative titration of the resulting nanoparticles with a cationic species (-NR4(+)) in a toluene phase, yielding ion pairing of -NR4(+) and -COO(-) on part of the capping monolayer. Analysis of the phase extraction allowed a quantitative determination of the percentage of ion pairing and structural changes in the capping monolayer on the nanoparticles. The results, along with morphological characterization, are discussed in terms of the interfacial structural changes and their implications on the rational design of surface-functionalized nanoparticles and fine tuning of the interfacial reactivity.

  4. Comparison of polarimetry and crown ether-based HPLC chiral stationary phase method to determine (L)-amino acid optical purity.

    PubMed

    Lee, Mee Sung; Khan, F Nawaz; Shin, Sung Chul; Jeong, Euh Duck; Kim, Hae Gyeong; Kim, Mi Jin; Cho, Yoon Jae; Cha, Jae-Young; Hyun, Myung Ho; Jin, Jong Sung

    2012-11-15

    Although various pharmacopoeias provide titration methods to assay (L)-amino acid content, none of these methods distinguish between (L)- and (D)-amino acids and do not consider the presence of enantiomeric impurities. Consequently, these methods are limited in scope to describe the relationship between content and specific rotation, [α]. In this study, the US Pharmacopoeia method was compared with the crown ether-based high performance liquid chromatographic (HPLC) chiral stationary phase (CSP) method to determine (L)-amino acid content and specific rotation. The (L)-amino acid content specified by the US Pharmacopoeia method was not consistent with the specific rotation in the presence of enantiomeric impurities, whereas the HPLC-CSP method was very effective for determining the (L)-amino acid content and the optical purity. The other advantage is that the HPLC-CSP method requires amino acid samples of quite low concentration (as low as 1 μg/mL), whereas the pharmacopoeia method requires higher concentrations (20-110 mg/mL).

  5. Comparison of polarimetry and crown ether-based HPLC chiral stationary phase method to determine (L)-amino acid optical purity.

    PubMed

    Lee, Mee Sung; Khan, F Nawaz; Shin, Sung Chul; Jeong, Euh Duck; Kim, Hae Gyeong; Kim, Mi Jin; Cho, Yoon Jae; Cha, Jae-Young; Hyun, Myung Ho; Jin, Jong Sung

    2012-11-15

    Although various pharmacopoeias provide titration methods to assay (L)-amino acid content, none of these methods distinguish between (L)- and (D)-amino acids and do not consider the presence of enantiomeric impurities. Consequently, these methods are limited in scope to describe the relationship between content and specific rotation, [α]. In this study, the US Pharmacopoeia method was compared with the crown ether-based high performance liquid chromatographic (HPLC) chiral stationary phase (CSP) method to determine (L)-amino acid content and specific rotation. The (L)-amino acid content specified by the US Pharmacopoeia method was not consistent with the specific rotation in the presence of enantiomeric impurities, whereas the HPLC-CSP method was very effective for determining the (L)-amino acid content and the optical purity. The other advantage is that the HPLC-CSP method requires amino acid samples of quite low concentration (as low as 1 μg/mL), whereas the pharmacopoeia method requires higher concentrations (20-110 mg/mL). PMID:22868097

  6. Method for the separation of acid from acid-laden vapors

    SciTech Connect

    Hansen, L.J.

    1992-02-11

    This patent describes a method for the removal of hydrochloric or sulfuric acid from vapor laden with the acid. It comprises: contacting the acid-laden vapors with packing materials in a zone containing the packing materials wherein the packing materials are formed of polyester resin containing from about 5 to 40 weight percent aluminum sulfate crystals.

  7. Boron containing amino acid compounds and methods for their use

    SciTech Connect

    Glass, J.D.; Coderre, J.A.

    2000-01-25

    The present invention provides new boron containing amino acid compounds and methods for making these compounds by contacting melphalan or another nitrogen mustard derivative and sodium borocaptate. The present invention also provides a method of treating a mammal having a tumor by administering to the mammal a therapeutically effective amount of the new boron containing amino acid compounds.

  8. Boron containing amino acid compounds and methods for their use

    DOEpatents

    Glass, John D.; Coderre, Jeffrey A.

    2000-01-01

    The present invention provides new boron containing amino acid compounds and methods for making these compounds by contacting melphalan or another nitrogen mustard derivative and sodium borocaptate. The present invention also provides a method of treating a mammal having a tumor by administering to the mammal a therapeutically effective amount of the new boron containing amino acid compounds.

  9. The standardization of infectivity titrations of poliovaccines--a WHO collaborative study.

    PubMed

    Magrath, D I; Seagroatt, V

    1985-04-01

    The reproducibility of a method for infectivity titrations of live poliovaccines using microtitre plates was investigated in a WHO collaborative study involving eight laboratories. The large variation (up to 100-fold) in estimates of infectivity observed between laboratories using their local methods of assay was reduced to no more than fourfold when a common method was used. However, expressing infectivities relative to those of the monovalent reference viruses improved the agreement between the laboratories irrespective of the titration method employed. On the basis of these results, WHO adopted the common method used in the study as its recommended method for poliovirus titrations and established the preparations studied as international reference materials for the infectivity titrations of live poliovaccines.

  10. Mutant fatty acid desaturase and methods for directed mutagenesis

    DOEpatents

    Shanklin, John; Whittle, Edward J.

    2008-01-29

    The present invention relates to methods for producing fatty acid desaturase mutants having a substantially increased activity towards substrates with fewer than 18 carbon atom chains relative to an unmutagenized precursor desaturase having an 18 carbon chain length specificity, the sequences encoding the desaturases and to the desaturases that are produced by the methods. The present invention further relates to a method for altering a function of a protein, including a fatty acid desaturase, through directed mutagenesis involving identifying candidate amino acid residues, producing a library of mutants of the protein by simultaneously randomizing all amino acid candidates, and selecting for mutants which exhibit the desired alteration of function. Candidate amino acids are identified by a combination of methods. Enzymatic, binding, structural and other functions of proteins can be altered by the method.

  11. Method for Enzyme Design with Genetically Encoded Unnatural Amino Acids.

    PubMed

    Hu, C; Wang, J

    2016-01-01

    We describe the methodologies for the design of artificial enzymes with genetically encoded unnatural amino acids. Genetically encoded unnatural amino acids offer great promise for constructing artificial enzymes with novel activities. In our studies, the designs of artificial enzyme were divided into two steps. First, we considered the unnatural amino acids and the protein scaffold separately. The scaffold is designed by traditional protein design methods. The unnatural amino acids are inspired by natural structure and organic chemistry methods, and synthesized by either organic chemistry methods or enzymatic conversion. With the increasing number of published unnatural amino acids with various functions, we described an unnatural amino acids toolkit containing metal chelators, redox mediators, and click chemistry reagents. These efforts enable a researcher to search the toolkit for appropriate unnatural amino acids for the study, rather than design and synthesize the unnatural amino acids from the beginning. After the first step, the model enzyme was optimized by computational methods and directed evolution. Lastly, we describe a general method for evolving aminoacyl-tRNA synthetase and expressing unnatural amino acids incorporated into a protein. PMID:27586330

  12. Method for Enzyme Design with Genetically Encoded Unnatural Amino Acids.

    PubMed

    Hu, C; Wang, J

    2016-01-01

    We describe the methodologies for the design of artificial enzymes with genetically encoded unnatural amino acids. Genetically encoded unnatural amino acids offer great promise for constructing artificial enzymes with novel activities. In our studies, the designs of artificial enzyme were divided into two steps. First, we considered the unnatural amino acids and the protein scaffold separately. The scaffold is designed by traditional protein design methods. The unnatural amino acids are inspired by natural structure and organic chemistry methods, and synthesized by either organic chemistry methods or enzymatic conversion. With the increasing number of published unnatural amino acids with various functions, we described an unnatural amino acids toolkit containing metal chelators, redox mediators, and click chemistry reagents. These efforts enable a researcher to search the toolkit for appropriate unnatural amino acids for the study, rather than design and synthesize the unnatural amino acids from the beginning. After the first step, the model enzyme was optimized by computational methods and directed evolution. Lastly, we describe a general method for evolving aminoacyl-tRNA synthetase and expressing unnatural amino acids incorporated into a protein.

  13. Gallic Acid: Review of the Methods of Determination and Quantification.

    PubMed

    Fernandes, Felipe Hugo Alencar; Salgado, Hérida Regina Nunes

    2016-05-01

    Gallic acid (3,4,5 trihydroxybenzoic acid) is a secondary metabolite present in most plants. This metabolite is known to exhibit a range of bioactivities including antioxidant, antimicrobial, anti-inflammatory, and anticancer. There are various methods to analyze gallic acid including spectrometry, chromatography, and capillary electrophoresis, among others. They have been developed to identify and quantify this active ingredient in most biological matrices. The aim of this article is to review the available information on analytical methods for gallic acid, as well as presenting the advantages and limitations of each technique.

  14. Initial Evaluation of a Titration Appliance for Temporary Treatment of Obstructive Sleep Apnea

    PubMed Central

    Levendowski, Daniel J.; Morgan, Todd; Westbrook, Philip

    2015-01-01

    Background Custom oral appliances that adjustably advance the mandible provide superior outcomes when treating patients with moderate or severe sleep apnea. Custom appliances, however, are expensive, must be fitted by a dentist, and the likelihood of successful outcomes are difficult to predict. An inexpensive trial appliance, if proven efficacious, might be used to predict custom appliance outcomes or to provide temporary therapeutic benefit. Objective The aim of this initial study was to assess the treatment efficacy of a novel titration oral appliance with that of an optimized custom appliance. Methods Seventeen patients, treated with a custom oral appliance for at least one year, successfully completed a three-night home sleep test. The baseline obstructive sleep apnea severity was established on Night 1 with seven patients exhibiting severe, six moderate and four mild apnea/hypopnea indexes. Patients were randomly assigned to wear their custom appliance or the titration appliance on Nights 2 and 3. Results Significant reductions in the mean overall and supine apnea indexes (p < 0.05), and the overall (p < 0.01) and supine (p < 0.05) apnea/hypopnea indexes were observed for both the titration and custom appliances. The proportion of patients who exhibited at least a 50% reduction in the overall apnea index and supine apnea/hypopnea were similar for the titration and custom appliance (~60%). The custom appliance reduced the overall apnea/hypopnea index by 50% in a greater proportion of the patients compared to the titration appliance (77% vs. 53%). The titration appliance significantly reduced the degree of hypoxic exposure across sleep disordered breathing events overall (p < 0.05) and supine (p < 0.01). Patients found their custom appliance was more comfortable than the titration appliance, but preferred the titration appliance to no therapy. Conclusion The titration appliance may be useful in assessing oral appliance treatment efficacy. When set to 70% of

  15. A hot-cell titration system

    SciTech Connect

    Klatt, L.N.

    1988-07-01

    Operation of nuclear fuel reprocessing plant requires an analytical support laboratory capable of meeting the process control, product quality, and nuclear safeguard requirements. Because of the radioactivity accompanying many of the samples, the analytical instruments must be selected, modified, or specifically developed for use in hot cells. Titrimetric procedures have been successfully used in hot cells and are generally immune to radiation induced bias. This report describes a titration system designed for operation in a hot-cell environment. The potentiometric titration system has operated successfully for four years in support of nuclear fuel reprocessing research and development activities. Details of the hardware, electronic, and software control and data analysis systems are presented. Interchangeable burets with a capacity of 5, 10, and 25 mL are available; the means of the absolute error in delivered volume for these burets are 0.9, 1.1, and 1.8 ..mu..L, respectively. Results of evaluation studies how that the accuracy and precision of analysis results obtained with the potentiometric system are limited by statistical uncertainties associated with the standard titrant, sample preparation procedure, and the equilibrium constant of the titration reaction and not by titrator performance factors. The system is also capable of performing amperometric titrations. Changing between the potentiometric and amperometric modes of operation involves changing the in-cell transducers, the in-cell electronics, and the titrator control program. 22 refs., 13 figs., 9 tabs.

  16. Subsite binding energies of an exo-polygalacturonase using isothermal titration calorimetry

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Thermodynamic parameters for binding of a series of galacturonic acid oligomers to an exo-polygalacturonase, RPG16 from Rhizopus oryzae, were determined by isothermal titration calorimetry. Binding of oligomers varying in chain length from two to five galacturonic acid residues is an exothermic proc...

  17. Compositions and method for controlling precipitation when acidizing sour wells

    SciTech Connect

    Dill, W.R.; Walker, M.L.

    1990-08-21

    This patent describes a method of treating a sour well penetrating a subterranean formation. It comprises: introducing into the well a treating fluid comprising an acid solution having a pH below 1.9, an iron sequestering agent comprising at least one compound selected from the group consisting of aminopolycarboxylic acids, hydroxycarboxylic acids, cyclic polyethers and derivatives of the acids and ethers, present in an amount of from about 0.25 to about 5 percent by weight of the acid solution, and a sulfide modifier comprising at least one compound selected from the group consisting of an aldehyde, acetal, hemiacetal and any other compound capable of forming aldehydes in the acid solution, present in an amount of from about 0.25 to about 5 percent of the acid solution; and treating the subterranean formation with the treating fluid.

  18. Influence of AgCl precipitates on the precipitation titration of sodium chloride by constant-current coulometry.

    PubMed

    Asakai, Toshiaki; Hara, Hisao; Murayama, Mariko; Tanaka, Tatsuhiko

    2006-08-01

    The precipitation titration of sodium chloride with electrogenerated silver ion was studied. The production of a precipitate of silver chloride had a significant effect on the titration results because the precipitate involved unreacted chloride or unreacted silver ion. The accuracy of the method was investigated by changing the introduction time of a sodium chloride solution to the coulometric cell during the process of electrolysis, and examining the dependency on the sample size. The accuracy of the measurement of the precipitation titration is discussed.

  19. Titration of adenovirus by counting cells containing virus-induced inclusion bodies.

    PubMed

    Weber, J

    1972-05-01

    A new method for the titration of adenovirus types 2 and 12 based on the enumeration of viral inclusions in infected cells was devised and evaluated. The technique gave virus titers comparable to those obtained by the plaque assay procedure.

  20. Volumetric determination of uranium using titanous sulfate as reductant before oxidimetric titration

    USGS Publications Warehouse

    Wahlberg, James S.; Skinner, Dwight L.; Rader, Lewis F.

    1956-01-01

    A new method for determining uranium in samples containing 0.05 percent or more U3O8, using titanous sulfate as reducing agent, is much shorter, faster, and has fewer interferences than conventional methods using reductor columns. The sample is dissolved with sulfuric, nitric, perchloric, and hydrofluoric acids. Elements that would otherwise form insoluble fluorides are kept in solution by complexing the fluoride ion with boric acid. A precipitation is made with cupferron to remove interfering elements. The solution is filtered to remove the precipitated cupferrates instead of extracting them with chloroform as is usually done. Filtration is preferred to extraction because any niobium that may be in solution forms an insoluble cupferrate that may be removed by filtering but is very difficult to extract with chloroform. Excess cupferron is destroyed by oxidizing with nitric and perchloric acids, and evaporating to dense fumes of sulfuric acid. The uranium is reduced to U(IV) by the addition of titanous sulfate, with cupric sulfate used as an indicator of the completeness of the reduction. Metallic copper is formed when all the uranium is reduced. The reduced copper is then reoxidized by the addition of mercuric perchlorate, an excess of ferric sulfate added, and the solution titrated immediately with standard ceric sulfate with ferroin as an indicator. Precision of the method compared favorable with methods in common use, both for uranium ores and for most types of uranium-rich materials.

  1. High-throughput titration of luciferase-expressing recombinant viruses.

    PubMed

    Garcia, Vanessa; Krishnan, Ramya; Davis, Colin; Batenchuk, Cory; Le Boeuf, Fabrice; Abdelbary, Hesham; Diallo, Jean-Simon

    2014-01-01

    Standard plaque assays to determine infectious viral titers can be time consuming, are not amenable to a high volume of samples, and cannot be done with viruses that do not form plaques. As an alternative to plaque assays, we have developed a high-throughput titration method that allows for the simultaneous titration of a high volume of samples in a single day. This approach involves infection of the samples with a Firefly luciferase tagged virus, transfer of the infected samples onto an appropriate permissive cell line, subsequent addition of luciferin, reading of plates in order to obtain luminescence readings, and finally the conversion from luminescence to viral titers. The assessment of cytotoxicity using a metabolic viability dye can be easily incorporated in the workflow in parallel and provide valuable information in the context of a drug screen. This technique provides a reliable, high-throughput method to determine viral titers as an alternative to a standard plaque assay.

  2. Human jagged polypeptide, encoding nucleic acids and methods of use

    DOEpatents

    Li, Linheng; Hood, Leroy

    2000-01-01

    The present invention provides an isolated polypeptide exhibiting substantially the same amino acid sequence as JAGGED, or an active fragment thereof, provided that the polypeptide does not have the amino acid sequence of SEQ ID NO:5 or SEQ ID NO:6. The invention further provides an isolated nucleic acid molecule containing a nucleotide sequence encoding substantially the same amino acid sequence as JAGGED, or an active fragment thereof, provided that the nucleotide sequence does not encode the amino acid sequence of SEQ ID NO:5 or SEQ ID NO:6. Also provided herein is a method of inhibiting differentiation of hematopoietic progenitor cells by contacting the progenitor cells with an isolated JAGGED polypeptide, or active fragment thereof. The invention additionally provides a method of diagnosing Alagille Syndrome in an individual. The method consists of detecting an Alagille Syndrome disease-associated mutation linked to a JAGGED locus.

  3. Method for incorporating radioactive phosphoric acid solutions in concrete

    DOEpatents

    Wolf, Gary A [Kennewick, WA; Smith, Jeffrey W [Lancaster, OH; Ihle, Nathan C [Walla Walla, WA

    1984-01-01

    A method for incorporating radioactive phosphoric acid solutions in concrete is described wherein the phosphoric acid is reacted with Ca(OH).sub.2 to form a precipitate of hydroxyapatite and the hydroxyapatite is mixed with portland cement to form concrete.

  4. Method for incorporating radioactive phosphoric acid solutions in concrete

    DOEpatents

    Wolf, G.A.; Smith, J.W.; Ihle, N.C.

    1982-07-08

    A method for incorporating radioactive phosphoric acid solutions in concrete is described wherein the phosphoric acid is reacted with Ca(OH)/sub 2/ to form a precipitate of hydroxyapatite and the hydroxyapatite is mixed with Portland cement to form concrete.

  5. [Use of coulometric titration for elucidating the mechanism of the oxidation of 6-APA alkaline breakdown products by halogens].

    PubMed

    Kharlamov, V T; Inkin, A A; Ermolina, G E

    1975-02-01

    Penaldinic acid and penicillamine were formed on alkali decomposition (1 N NaOH) of 6-APA for 20 minutes at room temperature, penicillamine being completely oxidized to disulphide by the air oxygen. Coulometric titration of the alkali decomposition products showed that generated chlorine in 0.5 N HCl solution or bromine in a week acid solution of KBr oxidized them with participation of 7 electrones. Generated iodine did not practically oxidize the 6-APA decomposition products during the coulometric titration.

  6. Quantitative real-time PCR for rapid and accurate titration of recombinant baculovirus particles.

    PubMed

    Hitchman, Richard B; Siaterli, Evangelia A; Nixon, Clare P; King, Linda A

    2007-03-01

    We describe the use of quantitative PCR (QPCR) to titer recombinant baculoviruses. Custom primers and probe were designed to gp64 and used to calculate a standard curve of QPCR derived titers from dilutions of a previously titrated baculovirus stock. Each dilution was titrated by both plaque assay and QPCR, producing a consistent and reproducible inverse relationship between C(T) and plaque forming units per milliliter. No significant difference was observed between titers produced by QPCR and plaque assay for 12 recombinant viruses, confirming the validity of this technique as a rapid and accurate method of baculovirus titration.

  7. Determination of zirconium by amperometric titration of excess complexone III with bismuth nitrate at a bismuth electrode

    SciTech Connect

    Kozulina, M.M.; Lepin, Y.K.; Songina, O.A.

    1985-10-01

    Among the various methods for determining zirconium is the amperometric titration of excess EDTA with bismuth nitrate. Such a titration was first used with a dropping mercury electrode. Here the authors investigate the conditions for titrating with a bismuth indicator electrode because a number of difficulties -current oscillation, mercury toxicity -- arise in work with the dropping mercury electrode. It is determined that the bismuth indicator electrode can in fact be used to determine zirconium by inverse amperometric titration of excess EDTA with bismuth nitrate.

  8. Copper and cobalt complexes of octadentate azamacrocycles: spectrophotometric titration, stopped-flow kinetics and crystallographic study.

    PubMed

    Ozay, Hava; Baran, Yakup; Ishii, Youichi

    2011-12-01

    Details of complex formation kinetics are reported for tetrakis(2-hydroxyethyl) substituted cyclen (L(1)) and cyclam (L(2)) with Cu(II) and Co(II). Stopped-flow kinetics and spectroscopic titration methods were employed for the activation parameters and stability constants, respectively. X-ray studies revealed that the pendant 2-hydroxyethyl groups are not equivalent: two are folded over the macrocycle and maintained by intramolecular hydrogen bonds while the others are extended and pointed away from the macrocyclic cavity. Complex formation kinetics and spectroscopic titration were performed in aqueous acidic buffer solutions. Thermodynamic and kinetic parameters revealed that the ring size of the macrocycles plays an extremely important role for each metal ion studied. Stopped-flow kinetic measurements explained the mechanism of the complex formation process of both Cu(II) and Co(II) which proceed in outer-sphere interactions with ligands. There are two steps in the complex formation of the system studied. The initial step is a second order reaction between the metal ion and macrocycle with a second order rate constant.

  9. Methods and compositions for efficient nucleic acid sequencing

    DOEpatents

    Drmanac, Radoje

    2002-01-01

    Disclosed are novel methods and compositions for rapid and highly efficient nucleic acid sequencing based upon hybridization with two sets of small oligonucleotide probes of known sequences. Extremely large nucleic acid molecules, including chromosomes and non-amplified RNA, may be sequenced without prior cloning or subcloning steps. The methods of the invention also solve various current problems associated with sequencing technology such as, for example, high noise to signal ratios and difficult discrimination, attaching many nucleic acid fragments to a surface, preparing many, longer or more complex probes and labelling more species.

  10. Methods and compositions for efficient nucleic acid sequencing

    DOEpatents

    Drmanac, Radoje

    2006-07-04

    Disclosed are novel methods and compositions for rapid and highly efficient nucleic acid sequencing based upon hybridization with two sets of small oligonucleotide probes of known sequences. Extremely large nucleic acid molecules, including chromosomes and non-amplified RNA, may be sequenced without prior cloning or subcloning steps. The methods of the invention also solve various current problems associated with sequencing technology such as, for example, high noise to signal ratios and difficult discrimination, attaching many nucleic acid fragments to a surface, preparing many, longer or more complex probes and labelling more species.

  11. Unusual properties of crocodilian ovomacroglobulin shown in its methylamine treatment and sulfhydryl titration

    SciTech Connect

    Arakawa, H.; Osada, T.; Ikai, A.

    1986-02-01

    The inhibitory activity of chicken and crocodilian ovomacroglobulins against trypsin was measured before and after their incubation with methylamine. The result for crocodilian ovomacroglobulin showed that methylamine treatment destroyed half of its activity, in unique contrast to human alpha 2-macroglobulin and chicken ovomacroglobulin for which methylamine either destroys the inhibitory activity of the former completely or does not affect that of the latter at all. Free sulfhydryl groups of chicken and crocodilian ovomacroglobulins were titrated with 5,5'-dithiobis(2-nitrobenzoic acid) before and after incubation with trypsin. Prior to the incubation with trypsin the chicken and crocodilian proteins respectively had 0 and 1 titratable sulfhydryl per molecule of Mr 720,000. After treatment with trypsin the crocodilian protein had 3.5-4 titratable sulfhydryls, whereas there were no titratable sulfhydryls in the chicken protein. After denaturation of the crocodilian protein in sodium dodecyl sulfate at 100 degrees C the number of titratable sulfhydryls was 4. Chicken ovomacroglobulin again did not have an appreciable number of titratable sulfhydryls under similar denaturing conditions. Incubation of crocodilian protein with (14C)methylamine showed an incorporation of at least 2 mol of methylamine per molecule. The result indicated the presence of three intramolecular thiol ester bonds in crocodilian ovomacroglobulin with differential stability against external perturbations.

  12. Fatty acids determination in Bronte pistachios by gas chromatographic method.

    PubMed

    Pantano, Licia; Lo Cascio, Giovanni; Alongi, Angelina; Cammilleri, Gaetano; Vella, Antonio; Macaluso, Andrea; Cicero, Nicola; Migliazzo, Aldo; Ferrantelli, Vincenzo

    2016-10-01

    A gas chromatographic with flame ionization detector (GC-MS FID) method for the identification and quantification of fatty acids based on the extraction of lipids and derivatisation of free acids to form methyl esters was developed and validated. The proposed method was evaluated to a number of standard FAs, and Bronte pistachios samples were used for that purpose and to demonstrate the applicability of the proposed method. In this regard, repeatability, mean and standard deviation of the analytical procedure were calculated. The results obtained have demonstrated oleic acid as the main component of Bronte pistachios (72.2%) followed by linoleic acid (13.4%) and showed some differences in composition with respect to Tunisian, Turkish and Iranian pistachios.

  13. Fatty acids determination in Bronte pistachios by gas chromatographic method.

    PubMed

    Pantano, Licia; Lo Cascio, Giovanni; Alongi, Angelina; Cammilleri, Gaetano; Vella, Antonio; Macaluso, Andrea; Cicero, Nicola; Migliazzo, Aldo; Ferrantelli, Vincenzo

    2016-10-01

    A gas chromatographic with flame ionization detector (GC-MS FID) method for the identification and quantification of fatty acids based on the extraction of lipids and derivatisation of free acids to form methyl esters was developed and validated. The proposed method was evaluated to a number of standard FAs, and Bronte pistachios samples were used for that purpose and to demonstrate the applicability of the proposed method. In this regard, repeatability, mean and standard deviation of the analytical procedure were calculated. The results obtained have demonstrated oleic acid as the main component of Bronte pistachios (72.2%) followed by linoleic acid (13.4%) and showed some differences in composition with respect to Tunisian, Turkish and Iranian pistachios. PMID:27265004

  14. Comparison of VFA titration procedures used for monitoring the biogas process.

    PubMed

    Lützhøft, Hans-Christian Holten; Boe, Kanokwan; Fang, Cheng; Angelidaki, Irini

    2014-05-01

    Titrimetric determination of volatile fatty acids (VFAs) contents is a common way to monitor a biogas process. However, digested manure from co-digestion biogas plants has a complex matrix with high concentrations of interfering components, resulting in varying results when using different titration procedures. Currently, no standardized procedure is used and it is therefore difficult to compare the performance among plants. The aim of this study was to evaluate four titration procedures (for determination of VFA-levels of digested manure samples) and compare results with gas chromatographic (GC) analysis. Two of the procedures are commonly used in biogas plants and two are discussed in literature. The results showed that the optimal titration results were obtained when 40 mL of four times diluted digested manure was gently stirred (200 rpm). Results from samples with different VFA concentrations (1-11 g/L) showed linear correlation between titration results and GC measurements. However, determination of VFA by titration generally overestimated the VFA contents compared with GC measurements when samples had low VFA concentrations, i.e. around 1 g/L. The accuracy of titration increased when samples had high VFA concentrations, i.e. around 5 g/L. It was further found that the studied ionisable interfering components had lowest effect on titration when the sample had high VFA concentration. In contrast, bicarbonate, phosphate and lactate had significant effect on titration accuracy at low VFA concentration. An extended 5-point titration procedure with pH correction was best to handle interferences from bicarbonate, phosphate and lactate at low VFA concentrations. Contrary, the simplest titration procedure with only two pH end-points showed the highest accuracy among all titration procedures at high VFA concentrations. All in all, if the composition of the digested manure sample is not known, the procedure with only two pH end-points should be the procedure of

  15. Titration of the bacteriorhodopsin Schiff base involves titration of an additional protein residue.

    PubMed

    Zadok, Uri; Asato, Alfred E; Sheves, Mordechai

    2005-06-14

    The retinal protein protonated Schiff base linkage plays a key role in the function of bacteriorhodopsin (bR) as a light-driven proton pump. In the unphotolyzed pigment, the Schiff base (SB) is titrated with a pK(a) of approximately 13, but following light absorption, it experiences a decrease in the pK(a) and undergoes several alterations, including a deprotonation process. We have studied the SB titration using retinal analogues which have intrinsically lower pK(a)'s which allow for SB titrations over a much lower pH range. We found that above pH 9 the channel for the SB titration is perturbed, and the titration rate is considerably reduced. On the basis of studies with several mutants, it is suggested that the protonation state of residue Glu204 is responsible for the channel perturbation. We suggest that above pH 12 a channel for the SB titration is restored probably due to titration of an additional protein residue. The observations may imply that during the bR photocycle and M photointermediate formation the rate of Schiff base protonation from the bulk is decreased. This rate decrease may be due to the deprotonation process of the "proton-releasing complex" which includes Glu204. In contrast, during the lifetime of the O intermediate, the protonated SB is exposed to the bulk. Possible implications for the switch mechanism, and the directionality of the proton movement, are discussed.

  16. Accurate approach for determining fresh-water carbonate (H2CO3(*)) alkalinity, using a single H3PO4 titration point.

    PubMed

    Birnhack, Liat; Sabach, Sara; Lahav, Ori

    2012-10-15

    A new, simple and accurate method is introduced for determining H(2)CO(3)(*) alkalinity in fresh waters dominated by the carbonate weak-acid system. The method relies on a single H(3)PO(4) dosage and two pH readings (acidic pH value target: pH~4.0). The computation algorithm is based on the concept that the overall alkalinity mass of a solution does not change upon the addition of a non-proton-accepting species. The accuracy of the new method was assessed batch-wise with both synthetic and actual tap waters and the results were compared to those obtained from two widely used alkalinity analysis methods (titration to pH~4.5 and the Gran titration method). The experimental results, which were deliberately obtained with simple laboratory equipment (glass buret, general-purpose pH electrode, magnetic stirrer) proved the method to be as accurate as the conventional methods at a wide range of alkalinity values (20-400 mg L(-1) as CaCO(3)). Analysis of the relative error attained in the proposed method as a function of the target (acidic) pH showed that at the range 4.0

  17. The influence of mechanochemical modification on prevention of toxic ability of humic acids towards phenanthrene in aquatic environment

    NASA Astrophysics Data System (ADS)

    Shekhovtsova, N. S.; Maltseva, E. V.; Glyzina, T. S.; Ovchinnikova, I. S.

    2015-11-01

    The aim of the research work is to quantify interaction between phenanthrene with modified humic acids in aquatic environment. The changes in the structure and properties of humic acids after modifications were studied with 1H NMR spectroscopy and potentiometric titration methods. Our research demonstrates that the application of thiourea as a modified agent increases the binding capacity of humic acids towards phenanthrene.

  18. Method to produce succinic acid from raw hydrolysates

    DOEpatents

    Donnelly, Mark I.; Sanville-Millard, Cynthia Y.; Nghiem, Nhuan Phu

    2004-06-01

    A method for producing succinic acid from industrial-grade hydrolysates is provided, comprising supplying an organism that contains mutations for the genes ptsG, pflB, and ldhA, allowing said organism to accumulate biomass, and allowing said organism to metabolize the hydrolysate. Also provided is a bacteria mutant characterized in that it produces succinic acid from substrate contained in industrial-grade hydrolysate in a ratio of between 0.6:1 and 1.3:1 succinic acid to substrate.

  19. Determination of vanadium(V) by direct automatic potentiometric titration with EDTA using a chemically modified electrode as a potentiometric sensor.

    PubMed

    Quintar, S E; Santagata, J P; Cortinez, V A

    2005-10-15

    A chemically modified electrode (CME) was prepared and studied as a potentiometric sensor for the end-point detection in the automatic titration of vanadium(V) with EDTA. The CME was constructed with a paste prepared by mixing spectral-grade graphite powder, Nujol oil and N-2-naphthoyl-N-p-tolylhydroxamic acid (NTHA). Buffer systems, pH effects and the concentration range were studied. Interference ions were separated by applying a liquid-liquid extraction procedure. The CME did not require any special conditioning before using. The electrode was constructed with very inexpensive materials and was easily made. It could be continuously used, at least two months without removing the paste. Automatic potentiometric titration curves were obtained for V(V) within 5 x 10(-5) to 2 x 10(-3)M with acceptable accuracy and precision. The developed method was applied to V(V) determination in alloys for hip prosthesis. PMID:18970248

  20. Method for distinctive estimation of stored acidity forms in acid mine wastes.

    PubMed

    Li, Jun; Kawashima, Nobuyuki; Fan, Rong; Schumann, Russell C; Gerson, Andrea R; Smart, Roger St C

    2014-10-01

    Jarosites and schwertmannite can be formed in the unsaturated oxidation zone of sulfide-containing mine waste rock and tailings together with ferrihydrite and goethite. They are also widely found in process wastes from electrometallurgical smelting and metal bioleaching and within drained coastal lowland soils (acid-sulfate soils). These secondary minerals can temporarily store acidity and metals or remove and immobilize contaminants through adsorption, coprecipitation, or structural incorporation, but release both acidity and toxic metals at pH above about 4. Therefore, they have significant relevance to environmental mineralogy through their role in controlling pollutant concentrations and dynamics in contaminated aqueous environments. Most importantly, they have widely different acid release rates at different pHs and strongly affect drainage water acidity dynamics. A procedure for estimation of the amounts of these different forms of nonsulfide stored acidity in mining wastes is required in order to predict acid release rates at any pH. A four-step extraction procedure to quantify jarosite and schwertmannite separately with various soluble sulfate salts has been developed and validated. Corrections to acid potentials and estimation of acid release rates can be reliably based on this method.

  1. Evaluation of acidity estimation methods for mine drainage, Pennsylvania, USA.

    PubMed

    Park, Daeryong; Park, Byungtae; Mendinsky, Justin J; Paksuchon, Benjaphon; Suhataikul, Ratda; Dempsey, Brian A; Cho, Yunchul

    2015-01-01

    Eighteen sites impacted by abandoned mine drainage (AMD) in Pennsylvania were sampled and measured for pH, acidity, alkalinity, metal ions, and sulfate. This study compared the accuracy of four acidity calculation methods with measured hot peroxide acidity and identified the most accurate calculation method for each site as a function of pH and sulfate concentration. Method E1 was the sum of proton and acidity based on total metal concentrations; method E2 added alkalinity; method E3 also accounted for aluminum speciation and temperature effects; and method E4 accounted for sulfate speciation. To evaluate errors between measured and predicted acidity, the Nash-Sutcliffe efficiency (NSE), the coefficient of determination (R (2)), and the root mean square error to standard deviation ratio (RSR) methods were applied. The error evaluation results show that E1, E2, E3, and E4 sites were most accurate at 0, 9, 4, and 5 of the sites, respectively. Sites where E2 was most accurate had pH greater than 4.0 and less than 400 mg/L of sulfate. Sites where E3 was most accurate had pH greater than 4.0 and sulfate greater than 400 mg/L with two exceptions. Sites where E4 was most accurate had pH less than 4.0 and more than 400 mg/L sulfate with one exception. The results indicate that acidity in AMD-affected streams can be accurately predicted by using pH, alkalinity, sulfate, Fe(II), Mn(II), and Al(III) concentrations in one or more of the identified equations, and that the appropriate equation for prediction can be selected based on pH and sulfate concentration. PMID:25399119

  2. Evaluation of acidity estimation methods for mine drainage, Pennsylvania, USA.

    PubMed

    Park, Daeryong; Park, Byungtae; Mendinsky, Justin J; Paksuchon, Benjaphon; Suhataikul, Ratda; Dempsey, Brian A; Cho, Yunchul

    2015-01-01

    Eighteen sites impacted by abandoned mine drainage (AMD) in Pennsylvania were sampled and measured for pH, acidity, alkalinity, metal ions, and sulfate. This study compared the accuracy of four acidity calculation methods with measured hot peroxide acidity and identified the most accurate calculation method for each site as a function of pH and sulfate concentration. Method E1 was the sum of proton and acidity based on total metal concentrations; method E2 added alkalinity; method E3 also accounted for aluminum speciation and temperature effects; and method E4 accounted for sulfate speciation. To evaluate errors between measured and predicted acidity, the Nash-Sutcliffe efficiency (NSE), the coefficient of determination (R (2)), and the root mean square error to standard deviation ratio (RSR) methods were applied. The error evaluation results show that E1, E2, E3, and E4 sites were most accurate at 0, 9, 4, and 5 of the sites, respectively. Sites where E2 was most accurate had pH greater than 4.0 and less than 400 mg/L of sulfate. Sites where E3 was most accurate had pH greater than 4.0 and sulfate greater than 400 mg/L with two exceptions. Sites where E4 was most accurate had pH less than 4.0 and more than 400 mg/L sulfate with one exception. The results indicate that acidity in AMD-affected streams can be accurately predicted by using pH, alkalinity, sulfate, Fe(II), Mn(II), and Al(III) concentrations in one or more of the identified equations, and that the appropriate equation for prediction can be selected based on pH and sulfate concentration.

  3. Colorimetric Titration Experiment for the Undergraduate Laboratory.

    ERIC Educational Resources Information Center

    Lopez, Edwin; Vassos, Basil H.

    1984-01-01

    Describes a colorimetric titration instrument usable in the undergraduate laboratory that fulfills the objectives of ruggedness, freedom from ambient light interference, and low cost. Although accessories can be added (raising the price), the basic instrument is low priced and can be used manually with a simple voltmeter. (JN)

  4. Isothermal Titration Calorimetry in the Student Laboratory

    ERIC Educational Resources Information Center

    Wadso, Lars; Li, Yujing; Li, Xi

    2011-01-01

    Isothermal titration calorimetry (ITC) is the measurement of the heat produced by the stepwise addition of one substance to another. It is a common experimental technique, for example, in pharmaceutical science, to measure equilibrium constants and reaction enthalpies. We describe a stirring device and an injection pump that can be used with a…

  5. Titrimetric and photometric methods for determination of hypochlorite in commercial bleaches.

    PubMed

    Jonnalagadda, Sreekanth B; Gengan, Prabhashini

    2010-01-01

    Two methods, simple titration and photometric methods for determination of hypochlorite are developed, based its reaction with hydrogen peroxide and titration of the residual peroxide by acidic permanganate. In the titration method, the residual hydrogen peroxide is estimated by titration with standard permanganate solution to estimate the hypochlorite concentration. The photometric method is devised to measure the concentration of remaining permanganate, after the reaction with residual hydrogen peroxide. It employs 4 ranges of calibration curves to enable the determination of hypochlorite accurately. The new photometric method measures hypochlorite in the range 1.90 x 10(-3) to 1.90 x 10(-2) M, with high accuracy and with low variance. The concentrations of hypochlorite in diverse commercial bleach samples and in seawater which is enriched with hypochlorite were estimated using the proposed method and compared with the arsenite method. The statistical analysis validates the superiority of the proposed method. PMID:20446147

  6. Methods And Devices For Characterizing Duplex Nucleic Acid Molecules

    DOEpatents

    Akeson, Mark; Vercoutere, Wenonah; Haussler, David; Winters-Hilt, Stephen

    2005-08-30

    Methods and devices are provided for characterizing a duplex nucleic acid, e.g., a duplex DNA molecule. In the subject methods, a fluid conducting medium that includes a duplex nucleic acid molecule is contacted with a nanopore under the influence of an applied electric field and the resulting changes in current through the nanopore caused by the duplex nucleic acid molecule are monitored. The observed changes in current through the nanopore are then employed as a set of data values to characterize the duplex nucleic acid, where the set of data values may be employed in raw form or manipulated, e.g., into a current blockade profile. Also provided are nanopore devices for practicing the subject methods, where the subject nanopore devices are characterized by the presence of an algorithm which directs a processing means to employ monitored changes in current through a nanopore to characterize a duplex nucleic acid molecule responsible for the current changes. The subject methods and devices find use in a variety of applications, including, among other applications, the identification of an analyte duplex DNA molecule in a sample, the specific base sequence at a single nulceotide polymorphism (SNP), and the sequencing of duplex DNA molecules.

  7. 40 CFR 425.03 - Sulfide analytical methods and applicability.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... Provisions § 425.03 Sulfide analytical methods and applicability. (a) The potassium ferricyanide titration... the potassium ferricyanide titration method for the determination of sulfide in wastewaters...

  8. 40 CFR 425.03 - Sulfide analytical methods and applicability.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... Provisions § 425.03 Sulfide analytical methods and applicability. (a) The potassium ferricyanide titration... the potassium ferricyanide titration method for the determination of sulfide in wastewaters...

  9. 40 CFR 425.03 - Sulfide analytical methods and applicability.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... Provisions § 425.03 Sulfide analytical methods and applicability. (a) The potassium ferricyanide titration... the potassium ferricyanide titration method for the determination of sulfide in wastewaters...

  10. Acid-base properties of water-soluble organic matter of forest soils, studied by the pK-spectroscopy method.

    PubMed

    Shamrikova, E V; Ryazanov, M A; Vanchikova, E V

    2006-11-01

    Using the potentiometric titration and pK spectroscopy method, acid-base properties of water-soluble organic matter of forest soils have been studied. Five acidic classes composed of different substances with pK(a) values around 3.6; 4.8; 6.7; 8.7 and 9.7 have been identified. Testing the properties of soluble soil fraction, it is to be taken into account that when it is isolated from non-soluble soil matter, some water-soluble substances remain in soil and do not pass into the solution. Most firmly adsorbed in soil are water-soluble components with pK(a) 9.6-9.8.

  11. Surfactant titration of nanoparticle-protein corona.

    PubMed

    Maiolo, Daniele; Bergese, Paolo; Mahon, Eugene; Dawson, Kenneth A; Monopoli, Marco P

    2014-12-16

    Nanoparticles (NP), when exposed to biological fluids, are coated by specific proteins that form the so-called protein corona. While some adsorbing proteins exchange with the surroundings on a short time scale, described as a "dynamic" corona, others with higher affinity and long-lived interaction with the NP surface form a "hard" corona (HC), which is believed to mediate NP interaction with cellular machineries. In-depth NP protein corona characterization is therefore a necessary step in understanding the relationship between surface layer structure and biological outcomes. In the present work, we evaluate the protein composition and stability over time and we systematically challenge the formed complexes with surfactants. Each challenge is characterized through different physicochemical measurements (dynamic light scattering, ζ-potential, and differential centrifugal sedimentation) alongside proteomic evaluation in titration type experiments (surfactant titration). 100 nm silicon oxide (Si) and 100 nm carboxylated polystyrene (PS-COOH) NPs cloaked by human plasma HC were titrated with 3-[(3-Cholamidopropyl) dimethylammonio]-1-propanesulfonate (CHAPS, zwitterionic), Triton X-100 (nonionic), sodium dodecyl sulfate (SDS, anionic), and dodecyltrimethylammonium bromide (DTAB, cationic) surfactants. Composition and density of HC together with size and ζ-potential of NP-HC complexes were tracked at each step after surfactant titration. Results on Si NP-HC complexes showed that SDS removes most of the HC, while DTAB induces NP agglomeration. Analogous results were obtained for PS NP-HC complexes. Interestingly, CHAPS and Triton X-100, thanks to similar surface binding preferences, enable selective extraction of apolipoprotein AI (ApoAI) from Si NP hard coronas, leaving unaltered the dispersion physicochemical properties. These findings indicate that surfactant titration can enable the study of NP-HC stability through surfactant variation and also selective separation

  12. Characterization of Sea Lettuce Surface Functional Groups by Potentiometric Titrations

    NASA Astrophysics Data System (ADS)

    Ebling, A. M.; Schijf, J.

    2008-12-01

    In pursuit of our ultimate goal to better understand the prodigious capacity of the marine macroalga Ulva lactuca (sea lettuce) for adsorbing a broad range of dissolved trace metals from seawater, we performed an initial characterization of its surface functional groups. Specifically, the number of distinct functional groups as well as their individual bulk concentrations and acid dissociation constants (pKas) were determined by potentiometric titrations in NaCl solutions of various ionic strengths (I = 0.01-5.0 M), under inert nitrogen atmosphere at 25°C. Depending on the ionic strength, Ulva samples were manually titrated down to pH 2 or 3 with 1 N HCl and then up to pH 10 with 1 N NaOH in steps of 0.1-0.2 units, continuously monitoring pH with a glass combination electrode. Titrations of a dehydrated Ulva standard reference material (BCR-279) were compared with fresh Ulva tissue cultured in our laboratory. A titration in filtered natural seawater was also compared with one in an NaCl solution of equal ionic strength. Equilibrium constants for the ionization of water in NaCl solutions as a function of ionic strength were obtained from the literature. Fits to the titration data ([H]T vs. pH) were performed with the FITEQL4.0 computer code using non-electrostatic 3-, 4-, and 5-site models, either by fixing ionic strength at its experimental value or by allowing it to be extrapolated to zero, while considering all functional group pKas and bulk concentrations as adjustable parameters. Since pKas and bulk concentrations were found to be strongly correlated, the latter were also fixed in some cases to further constrain the pKas. Whereas these calculations are currently ongoing, preliminary results point to three, possibly four, functional groups with pKas of about 4.1, 6.3, and 9.5 at I = 0. Bulk concentrations of the three groups are very similar, about 5-6×10-4 mol/g based on dry weight, which suggests that all are homogeneously distributed over the surface and

  13. A method to attenuate U(VI) mobility in acidic waste plumes using humic acids

    SciTech Connect

    Wan, J.; Dong, W.; Tokunaga, T.K.

    2011-02-01

    Acidic uranium (U) contaminated plumes have resulted from acid-extraction of plutonium during the Cold War and from U mining and milling operations. A sustainable method for in-situ immobilization of U under acidic conditions is not yet available. Here, we propose to use humic acids (HAs) for in-situ U immobilization in acidic waste plumes. Our laboratory batch experiments show that HA can adsorb onto aquifer sediments rapidly, strongly and practically irreversibly. Adding HA greatly enhanced U adsorption capacity to sediments at pH below 5.0. Our column experiments using historically contaminated sediments from the Savannah River Site under slow flow rates (120 and 12 m/y) show that desorption of U and HA were non-detectable over 100 pore-volumes of leaching with simulated acidic groundwaters. Upon HA-treatment, 99% of the contaminant [U] was immobilized at pH < 4.5, compared to 5% and 58% immobilized in the control columns at pH 3.5 and 4.5, respectively. These results demonstrated that HA-treatment is a promising in-situ remediation method for acidic U waste plumes. As a remediation reagent, HAs are resistant to biodegradation, cost effective, nontoxic, and easily introducible to the subsurface.

  14. Method to attenuate U(VI) mobility in acidic waste plumes using humic acids.

    PubMed

    Wan, Jiamin; Dong, Wenming; Tokunaga, Tetsu K

    2011-03-15

    Acidic uranium (U) groundwater plumes have resulted from acid-extraction of plutonium during the Cold War and from U mining and milling operations. A sustainable method for in situ immobilization of U under acidic conditions is not yet available. Here, we propose to use humic acids (HAs) for in situ U immobilization in acidic waste plumes. Our laboratory batch experiments show that HA can adsorb onto aquifer sediments rapidly, strongly and practically irreversibly. Adding HA greatly enhanced U adsorption capacity to sediments at pH below 5.0. Our column experiments using historically contaminated sediments from the Savannah River Site under slow flow rates (120 and 12 m/year) show that desorption of U and HA were nondetectable over 100 pore-volumes of leaching with simulated acidic groundwaters. Upon HA-treatment, 99% of the contaminant [U] was immobilized at pH ≤ 4.5, compared to 5% and 58% immobilized in the control columns at pH 3.5 and 4.5, respectively. These results indicate that HA-treatment is a promising in situ remediation method for acidic U waste plumes. As a remediation reagent, HAs are resistant to biodegradation, cost-effective, nontoxic, and easily introducible to the subsurface.

  15. Method to attenuate U(VI) mobility in acidic waste plumes using humic acids.

    PubMed

    Wan, Jiamin; Dong, Wenming; Tokunaga, Tetsu K

    2011-03-15

    Acidic uranium (U) groundwater plumes have resulted from acid-extraction of plutonium during the Cold War and from U mining and milling operations. A sustainable method for in situ immobilization of U under acidic conditions is not yet available. Here, we propose to use humic acids (HAs) for in situ U immobilization in acidic waste plumes. Our laboratory batch experiments show that HA can adsorb onto aquifer sediments rapidly, strongly and practically irreversibly. Adding HA greatly enhanced U adsorption capacity to sediments at pH below 5.0. Our column experiments using historically contaminated sediments from the Savannah River Site under slow flow rates (120 and 12 m/year) show that desorption of U and HA were nondetectable over 100 pore-volumes of leaching with simulated acidic groundwaters. Upon HA-treatment, 99% of the contaminant [U] was immobilized at pH ≤ 4.5, compared to 5% and 58% immobilized in the control columns at pH 3.5 and 4.5, respectively. These results indicate that HA-treatment is a promising in situ remediation method for acidic U waste plumes. As a remediation reagent, HAs are resistant to biodegradation, cost-effective, nontoxic, and easily introducible to the subsurface. PMID:21319737

  16. Handling of electronic absorption spectra with a desk-top computer-II: calculation of stability constants from spectrophotometric titrations.

    PubMed

    Zuberbühler, A D; Kaden, T A

    1979-12-01

    A fully automatic system for combined spectrophotometric and pH titrations was described in Part I. Its performance in the titration of weak acids and metal complexes is discussed, along with a computer program for numerical treatment of the data, based on Marquardt's modification of the Newton-Gauss non-linear least-squares method. The deprotonation of p-nitrophenol at concentrations of 4 x 10(-5) and 4 x 10(-6)M was studied in order to test the sensitivity. Results identical within the reproducibility of the pH-meter were obtained: pK(H) = 7.00 +/- 0.01 and 7.02 +/- 0.01, respectively. Three complexation reactions were studied: (1) the interaction of SCN(-) with the Co(2+) complex of 1,4,8,11-tetramethyl-1,4,8,11-tetra-azacyclotetradecane (TMC); five independent experiments gave pK [CoTMC (SCN)(+) right harpoon over left harpoon CoTMC(2+) + SCN(-)] = 3.099 +/- 0.003: (2) the deprotonation of the Cu(2+) complex of 3,7-diazanonanediamide (DANA); five experiments gave pK (CuDANA(2+) right harpoon over left harpoon CuDANAH(+)(-1) + H(+)) = 7.14 +/- 0.01 and pK (CuDANAH(+)(-1) right harpoon over left harpoon CuDANAH(-2) + H(+)) = 8.38 +/- 0.01: (3) for the reaction of Cu(2+) with 1,3,7-triazacyclodecane (L), data from different ligand: metal ratios had to be combined to obtain pK (CuL(2+) right harpoon over left harpoon Cu(2+) + L) = 16.19 +/- 0.01, pK (CuL(2+)(2) right harpoon over left harpoon CuL(2+) + L) = 10.30 +/- 0.01, and pK (Cu(2)L(2) (OH)(2+)(2) right harpoon over left harpoon 2 CuL(2+) + 2 OH(-)) = 14.58 +/- 0.03. Titration curves with a total change in absorbance of as little as 0.03 units could be analysed satisfactorily, extending considerably the useful range of concentrations for spectrophotometric titrations. In combined spectrophotometric/pH titrations the accuracy of the glass electrode is normally the limiting factor. Other equilibrium constants can easily be reproduced with standard errors of less than 0.01 log unit.

  17. Two methods for the separation of monounsaturated octadecenoic acid isomers.

    PubMed

    Villegas, C; Zhao, Y; Curtis, J M

    2010-01-29

    The identification and quantification of complex mixtures of cis and trans octadecenoic (18:1) fatty acid isomers presents a major challenge for conventional one-dimensional GC/FID analysis of their methyl esters. We have compared the use of two methods to achieve optimized separations of positional and geometrical octadecenoic fatty acid isomers-comprehensive two-dimensional gas chromatography (GCxGC), and silver ion high performance liquid chromatography interfaced to atmospheric pressure photoionization (APPI) mass spectrometry. Nine isomers of octadecenoic acid methyl ester were well separated on a single silver ion column with a mobile phase of 0.018% acetonitrile and 0.18% isopropanol in hexane. Reproducible retention times were obtained with relative standard deviations of around 1% over 5 injections. The extra selectivity and reproducibility afforded by APPI-MS, together with the wide separation of cis and trans isomers by silver ion chromatography, resulted in a promising method for measurement of octadecenoic acid FAME. The GCxGC separation was performed using various column combinations, and optimal separation was obtained by coupling an ionic liquid column (Supelco SLB-IL100 [1,9-di(3-vinyl-imidazolium) nonane bis(trifluoromethyl) sulfonyl imidate]) in the first dimension with a SGE BPX50 (50% phenyl polysilphenylene-siloxane) in the second dimension. These methods have been applied to the analysis of octadecenoic acid in milk and beef fat. PMID:20022011

  18. A GC-ECD method for estimation of free and bound amino acids, gamma-aminobutyric acid, salicylic acid, and acetyl salicylic acid from Solanum lycopersicum (L.).

    PubMed

    Meher, Hari Charan; Gajbhiye, Vijay T; Singh, Ghanendra

    2011-01-01

    A gas chromatograph with electron capture detection method for estimation of selected metabolites--amino acids (free and bound), gamma-aminobutyric acid (GABA), salicylic acid (SA), and acetyl salicylic acid (ASA) from tomato--is reported. The method is based on nitrophenylation of the metabolites by 1-fluoro-2, 4-dinitrobenzene under aqueous alkaline conditions to form dinitophenyl derivatives. The derivatives were stable under the operating conditions of GC. Analysis of bound amino acids comprised perchloric acid precipitation of protein, alkylation (carboxymethylation) with iodoacetic acid, vapor-phase hydrolysis, and derivatization with 1-fluoro-2,4-dinitrobenzene in that order. The metabolites were resolved in 35 min, using a temperature-programmed run. The method is rapid, sensitive, and precise. It easily measured the typical amino acids (aspartate, asparagine, glutamate, glutamine, alanine, leucine, lysine, and phenylalanine) used for identification and quantification of a protein, resolved amino acids of the same mass (leucine and isoleucine), satisfactorily measured sulfur amino acid (methionine, cystine, and cysteine), and quantified GABA, SA, and ASA, as well. The developed method was validated for specificity, linearity, and precision. It has been applied and recommended for estimation of 25 metabolites from Solanum lycopersicum (L.).

  19. A GC-ECD method for estimation of free and bound amino acids, gamma-aminobutyric acid, salicylic acid, and acetyl salicylic acid from Solanum lycopersicum (L.).

    PubMed

    Meher, Hari Charan; Gajbhiye, Vijay T; Singh, Ghanendra

    2011-01-01

    A gas chromatograph with electron capture detection method for estimation of selected metabolites--amino acids (free and bound), gamma-aminobutyric acid (GABA), salicylic acid (SA), and acetyl salicylic acid (ASA) from tomato--is reported. The method is based on nitrophenylation of the metabolites by 1-fluoro-2, 4-dinitrobenzene under aqueous alkaline conditions to form dinitophenyl derivatives. The derivatives were stable under the operating conditions of GC. Analysis of bound amino acids comprised perchloric acid precipitation of protein, alkylation (carboxymethylation) with iodoacetic acid, vapor-phase hydrolysis, and derivatization with 1-fluoro-2,4-dinitrobenzene in that order. The metabolites were resolved in 35 min, using a temperature-programmed run. The method is rapid, sensitive, and precise. It easily measured the typical amino acids (aspartate, asparagine, glutamate, glutamine, alanine, leucine, lysine, and phenylalanine) used for identification and quantification of a protein, resolved amino acids of the same mass (leucine and isoleucine), satisfactorily measured sulfur amino acid (methionine, cystine, and cysteine), and quantified GABA, SA, and ASA, as well. The developed method was validated for specificity, linearity, and precision. It has been applied and recommended for estimation of 25 metabolites from Solanum lycopersicum (L.). PMID:21391500

  20. A titration model for evaluating calcium hydroxide removal techniques

    PubMed Central

    PHILLIPS, Mark; McCLANAHAN, Scott; BOWLES, Walter

    2015-01-01

    Objective Calcium hydroxide (Ca(OH)2) has been used in endodontics as an intracanal medicament due to its antimicrobial effects and its ability to inactivate bacterial endotoxin. The inability to totally remove this intracanal medicament from the root canal system, however, may interfere with the setting of eugenol-based sealers or inhibit bonding of resin to dentin, thus presenting clinical challenges with endodontic treatment. This study used a chemical titration method to measure residual Ca(OH)2 left after different endodontic irrigation methods. Material and Methods Eighty-six human canine roots were prepared for obturation. Thirty teeth were filled with known but different amounts of Ca(OH)2 for 7 days, which were dissolved out and titrated to quantitate the residual Ca(OH)2 recovered from each root to produce a standard curve. Forty-eight of the remaining teeth were filled with equal amounts of Ca(OH)2 followed by gross Ca(OH)2 removal using hand files and randomized treatment of either: 1) Syringe irrigation; 2) Syringe irrigation with use of an apical file; 3) Syringe irrigation with added 30 s of passive ultrasonic irrigation (PUI), or 4) Syringe irrigation with apical file and PUI (n=12/group). Residual Ca(OH)2 was dissolved with glycerin and titrated to measure residual Ca(OH)2 left in the root. Results No method completely removed all residual Ca(OH)2. The addition of 30 s PUI with or without apical file use removed Ca(OH)2 significantly better than irrigation alone. Conclusions This technique allowed quantification of residual Ca(OH)2. The use of PUI (with or without apical file) resulted in significantly lower Ca(OH)2 residue compared to irrigation alone. PMID:25760272

  1. A rapid automated procedure for laboratory and shipboard spectrophotometric measurements of seawater alkalinity: continuously monitored single-step acid additions

    NASA Astrophysics Data System (ADS)

    Liu, X.; Byrne, R. H.; Lindemuth, M.; Easley, R. A.; Patsavas, M. C.

    2012-12-01

    An automated system for shipboard and laboratory alkalinity measurements is presented. The simple system, which consists of a Dosimat titrator to deliver acid volumetrically and a USB 4000 spectrophotometer to monitor the titration progress, provides fast, precise and accurate measurements of total alkalinity for oceanographic research. The analytical method is based on single-point HCl titrations of seawater samples of a known volume; bromol cresol purple is used as an indicator to determine the final pH. Field data from an Arctic cruise demonstrates accuracy and precision around 1 micro mol/kg and a sample processing rate of 6 min per sample.

  2. Method for preparing 6-.beta.-halopenicillanic acids

    DOEpatents

    Hansen, Erik I.; Kran-Nielsen, Mogens P.; Von Daehne, Welf

    1989-01-01

    The present invention relates to a new and improved method for the preparation of a compound of the formula I ##STR1## in which R stands for halogen, giving rise to high yields of substantially pure 6.beta.-halopenicillanic acids, obtained in one step.

  3. A simplified method for estimation of jarosite and acid-forming sulfates in acid mine wastes.

    PubMed

    Li, Jun; Smart, Roger St C; Schumann, Russell C; Gerson, Andrea R; Levay, George

    2007-02-01

    In acid base accounting (ABA) estimates of acid mine wastes, the acid potential (AP) estimate can be improved by using the net carbonate value (NCV) reactive sulfide S method rather than total S assay methods but this does not give recovery of potentially acid producing ferrous and ferric sulfates present in many wastes. For more accurate estimation of AP, an effective, site-specific method to quantify acid sulfate salts, such as jarosite and melanterite, in waste rocks has been developed and tested on synthetic and real wastes. The SPOCAS (acid sulfate soils) methods have been modified to an effective, rapid method to speciate sulfate forms in different synthetic waste samples. A three-step sequential extraction procedure has been established. These steps are: (1) argon-purged water extraction (3 min) to extract soluble Fe(II) salts (particularly melanterite), epsomite and gypsum (<10 wt.%), (2) roasting at 550 degrees C (1 h) to remove sulfur from pyrite and other reactive sulfides, (3) HCl extraction (4 M, 30 min) for determination of jarosites. Products (solid and aqueous) have been characterized at each step including the jarosite decomposition process in Step 2 where temperature control is critical to avoid S loss. The sequential extraction procedure was used to quantitatively determine melanterite, epsomite, gypsum, pyrite and jarosite concentrations in a synthetic waste sample containing these mineral phases at 5 wt.% in quartz, and also tested using a tailings waste sample to quantitatively determine epsomite, gypsum and jarosite contents. The method is applicable to most waste samples including those with non-pyrite sulfides but for samples containing significant amounts of sulfur (>1 wt.% S) as copper sulfides, the second step of roasting needs to be excluded from the procedure with an increased time of 4 M HCl extraction to 16 h for jarosite determination.

  4. pH titrations of molluscan paramyosin at two different ionic strengths.

    PubMed

    Cooley, L B; Krause, S

    1980-11-01

    Paramyosin extracted from the adductor muscle of Mercenaria mercenaria, the chowder clam, was titrated both in 0.3 M KCl and in 1 mM KCl. Both the presumed native form of the molecule, acid-R-paramyosin, and a slightly degraded form, beta-paramyosin, were studied. Titrations of both types of paramyosin were similar in 1 mM k+, except that the native paramyosin is more highly charged at pH 3.2 than beta-paramyosin, as postulated previously (DeLaney and Krause, 1976, Macromolecules, 9:455), and that more groups titrate on the native molecule than on beta-paramyosin, both between pH 3.2 and 3.3 and between pH 3.2 and 10. Titrations in 0.30 M KCl, unlike those in 1 mM K, depended on starting pH; long term exposure to alkali solutions during dialysis, previously shown to cause partial dephosphorylation of paramyosin (Cooley et al., 1979, J. Biol. Chem., 254:2195), apparently also leads to a change in intermolecular interactions sufficient to cause changes in the titration curves in 0.30 M KCl but not in 1 mM K+.

  5. pH titrations of molluscan paramyosin at two different ionic strengths.

    PubMed Central

    Cooley, L B; Krause, S

    1980-01-01

    Paramyosin extracted from the adductor muscle of Mercenaria mercenaria, the chowder clam, was titrated both in 0.3 M KCl and in 1 mM KCl. Both the presumed native form of the molecule, acid-R-paramyosin, and a slightly degraded form, beta-paramyosin, were studied. Titrations of both types of paramyosin were similar in 1 mM k+, except that the native paramyosin is more highly charged at pH 3.2 than beta-paramyosin, as postulated previously (DeLaney and Krause, 1976, Macromolecules, 9:455), and that more groups titrate on the native molecule than on beta-paramyosin, both between pH 3.2 and 3.3 and between pH 3.2 and 10. Titrations in 0.30 M KCl, unlike those in 1 mM K, depended on starting pH; long term exposure to alkali solutions during dialysis, previously shown to cause partial dephosphorylation of paramyosin (Cooley et al., 1979, J. Biol. Chem., 254:2195), apparently also leads to a change in intermolecular interactions sufficient to cause changes in the titration curves in 0.30 M KCl but not in 1 mM K+. PMID:7260299

  6. Determination of the phenolic-group capacities of humic substances by non-aqueous titration technique.

    PubMed

    Kirishima, Akira; Ohnishi, Takashi; Sato, Nobuaki; Tochiyama, Osamu

    2009-07-15

    The phenolic-group capacities of five humic substances, such as, the Aldrich humic acid, the humic and fulvic acids extracted from a soil, the humic and fulvic acids extracted from a peat have been precisely determined by the non-aqueous potentiometric titration technique. The titration by KOH in the mixed solvent of DMSO:2-propanol:water=80:19.3:0.7 at [K(+)]=0.02 M enabled to measure the potential change in a wide range of pOH (=-log[OH(-)]), and thus to determine the capacities of phenolic groups which could not be precisely determined in the aqueous titration. The results of the titration revealed that the mean protonation constants of the phenolic groups were nearly the same for all humic substances and close to that of phenol in the same medium, indicating that each phenolic-group in the humic substances is rather isolated and is not electronically affected by other affecting groups in the humic macromolecule.

  7. A Simple and Universal Method for Making up Buffer Solutions.

    ERIC Educational Resources Information Center

    Dennison, Clive

    1988-01-01

    Uses a method which involves weighing out an amount of the appropriate weak acid or base and dissolving it in distilled water close to the final volume. Solution is then titrated with strong acid or base to give the desired pH. Provides three examples. (MVL)

  8. High concentration aqueous sodium fluoride certified reference materials for forensic use certified by complexometric titration.

    PubMed

    Archer, Marcellé; Brits, Martin; Prevoo-Franzsen, Désirée; Quinn, Laura

    2015-04-01

    Sodium fluoride in concentrations of 1 to 2 % is used to prevent the formation of ethanol in blood and urine samples that are to be analysed for ethanol content. The majority of such samples form part of forensic investigations into alleged drunken driving. In South Africa, the laboratory performing the tests is required to prove that the sodium fluoride concentration in the blood samples is above 1 g/100 ml on receipt. This is done by using a fluoride ion-selective electrode calibrated with external aqueous solutions of sodium fluoride. The National Metrology Institute of South Africa (NMISA) prepares sodium fluoride solutions in concentrations from 0.3 to 3.0 g/100 ml. No other certified sodium fluoride reference solutions in these concentrations are available commercially. The sodium fluoride is certified by precipitation of the fluoride as lead chlorofluoride (PbClF) through the addition of a known excess of lead nitrate. The excess lead is back-titrated with ethylenediamine tetraacetic acid (EDTA) using a photometric electrode to detect the endpoint. Aqueous sodium fluoride solutions are prepared and the concentrations verified by the precipitation/back-titration method. This paper shows the application of a classical complexometric method to the certification of reference materials and describes the procedures for the preparation of the sodium fluoride solutions, verification of the concentrations, homogeneity and stability by primary titrimetry. It also briefly covers the calculation of uncertainty, the establishment of traceability and the quality control measures applied to ensure the quality of the certified reference materials (CRMs). PMID:25326884

  9. High concentration aqueous sodium fluoride certified reference materials for forensic use certified by complexometric titration.

    PubMed

    Archer, Marcellé; Brits, Martin; Prevoo-Franzsen, Désirée; Quinn, Laura

    2015-04-01

    Sodium fluoride in concentrations of 1 to 2 % is used to prevent the formation of ethanol in blood and urine samples that are to be analysed for ethanol content. The majority of such samples form part of forensic investigations into alleged drunken driving. In South Africa, the laboratory performing the tests is required to prove that the sodium fluoride concentration in the blood samples is above 1 g/100 ml on receipt. This is done by using a fluoride ion-selective electrode calibrated with external aqueous solutions of sodium fluoride. The National Metrology Institute of South Africa (NMISA) prepares sodium fluoride solutions in concentrations from 0.3 to 3.0 g/100 ml. No other certified sodium fluoride reference solutions in these concentrations are available commercially. The sodium fluoride is certified by precipitation of the fluoride as lead chlorofluoride (PbClF) through the addition of a known excess of lead nitrate. The excess lead is back-titrated with ethylenediamine tetraacetic acid (EDTA) using a photometric electrode to detect the endpoint. Aqueous sodium fluoride solutions are prepared and the concentrations verified by the precipitation/back-titration method. This paper shows the application of a classical complexometric method to the certification of reference materials and describes the procedures for the preparation of the sodium fluoride solutions, verification of the concentrations, homogeneity and stability by primary titrimetry. It also briefly covers the calculation of uncertainty, the establishment of traceability and the quality control measures applied to ensure the quality of the certified reference materials (CRMs).

  10. A monosegmented-flow Karl Fischer titrator.

    PubMed

    de Aquino, Emerson Vidal; Rohwedder, Jarbas José Rodrigues; Pasquini, Celio

    2007-02-28

    A monosegmented volumetric Karl Fischer titrator is described to mechanize the determination of water content in organic solvents. The system is based on the flow-batch characteristics of the monosegmented analysis concept and employs biamperometry to monitor the progress of the titration. The system shows accuracy and precision that are highly independent of the flow rate, does not require calibration, and is carried out in a closed system capable of minimizing contact of the sample and reagents with ambient moisture. Sample volumes in the range of 40-300muL are employed, depending on the water concentration. An automatic dilution is provided to deal with concentrated samples. The consumption of Karl Fischer reagent depends on the water content of the sample but is not larger than 100muL. The system was evaluated for determination of water in ethanol and methanol in the range 0.02-0.5% (w/w). The average relative precision estimated in that range (9-3%) is comparable to that obtained with a larger volume commercial system and no significant difference was observed between the results obtained for the two systems at the 95% confidence level. A complete titration can be performed in less than 5min employing the proposed system. PMID:19071447

  11. Acid pre-treatment method for in situ ore leaching

    DOEpatents

    Mallon, R.G.; Braun, R.L.

    1975-10-28

    An acid leaching method is described for the recovery of a desired element from a subterranean rubblized body of primary ore containing the element and also having associated therewith a carbonate mineral wherein the rubblized ore body is flooded with an aqueous acidic solution in order to release carbon dioxide from the associated carbonate mineral. After a substantial portion of the available carbon dioxide is released and removed from the ore body, as by venting to the atmosphere, an oxidizing gas is introduced into the flooded, rubblized ore to oxidize the ore and form an acid leach solution effective in the presence of the dissolved oxidizing gas to dissolve the ore and cause the desired element to go into solution. The leach solution is then circulated to the surface where the metal values are recovered therefrom.

  12. 40 CFR Appendix A to Subpart Ddd... - Free Formaldehyde Analysis of Insulation Resins by the Hydroxylamine Hydrochloride Method

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... this method is the titration of the hydrochloric acid that is liberated when hydroxylamine... prestandardized pH meter, 1.0 N hydrochloric acid, 0.1 N hydrochloric acid, and 0.1 N sodium hydroxide. 5.5Add...

  13. Combining pH and electrical conductivity measurements to improve titrimetric methods to determine ammonia nitrogen, volatile fatty acids and inorganic carbon concentrations.

    PubMed

    Charnier, C; Latrille, E; Lardon, L; Miroux, J; Steyer, J P

    2016-05-15

    Volatile fatty acids (VFA), inorganic carbon (IC) and total ammonia nitrogen (TAN) are key variables in the current context of anaerobic digestion (AD). Accurate measurements like gas chromatography and infrared spectrometry have been developed to follow the concentration of these compounds but none of these methods are affordable for small AD units. Only titration methods answer the need for small plant monitoring. The existing methods accuracy was assessed in this study and reveals a lack of accuracy and robustness to control AD plants. To solve these issues, a new titrimetric device to estimate the VFA, IC and TAN concentrations with an improved accuracy was developed. This device named SNAC (System of titration for total ammonia Nitrogen, volatile fatty Acids and inorganic Carbon) has been developed combining the measurement of electrical conductivity and pH. SNAC were tested on 24 different plant samples in a range of 0-0.16 mol.L(-1) TAN, 0.01-0.21 mol.L(-1) IC and 0-0.04 mol.L(-1) VFA. The standard error was about 0.012 mol.L(-1) TAN, 0.015 mol.L(-1) IC and 0.003 mol.L(-1) VFA. The coefficient of determination R(2) between the estimated and reference data was 0.95, 0.94 and 0.95 for TAN, IC and VFA respectively. Using the same data, current methods based on key pH points lead to standard error more than 14.5 times higher on VFA and more than 1.2 times higher on IC. These results show that SNAC is an accurate tool to improve the management of AD plant.

  14. Kinetic detection of the end-point in titrations involving slow reactions Direct titration of polyhydroxy-compounds with periodate.

    PubMed

    Efstathiou, C E; Hadjiioannou, T P

    1983-03-01

    A new titration technique is described in which the end-point is determined by measuring the relative reaction rate of the titration reaction. This technique is adequate for rather slow reactions where conventional direct titrations are not applicable. The titrations are done automatically under microcomputer control. The efficiency of this technique is demonstrated with direct titrations of certain polyhydroxy-compounds with standard periodate solution. Ethylene glycol and propylene glycol (0.05-0.3 mmole), glycerol (0.06-0.17 mmole) and mannitol (0.01-0.03 mmole) were determined with average relative errors of 0.1-0.3%.

  15. Method for liquid chromatographic extraction of strontium from acid solutions

    DOEpatents

    Horwitz, E. Philip; Dietz, Mark L.

    1992-01-01

    A method and apparatus for extracting strontium and technetium values from biological, industrial and environmental sample solutions using a chromatographic column is described. An extractant medium for the column is prepared by generating a solution of a diluent containing a Crown ether and dispersing the solution on a resin substrate material. The sample solution is highly acidic and is introduced directed to the chromatographic column and strontium or technetium is eluted using deionized water.

  16. Method and apparatus for staining immobilized nucleic acids

    DOEpatents

    Ramsey, J. Michael; Foote, Robert S.; Jacobson, Stephen C.

    2000-01-01

    A method for staining immobilized nucleic acids includes the steps of affixing DNA probes to a solid substrate, moving target DNA material into proximity with the DNA probes, whereby the target DNA hybridized with specific ones of the DNA probes, and moving a fluorescent dye into proximity with the hybridized target DNA, whereby the fluorescent dye binds to the hybridized DNA to enable subsequent detection of fluorescence.

  17. Preparation of polyaniline nanostructures doped with different dicarboxylic acids through template-free method

    NASA Astrophysics Data System (ADS)

    Sun, Chuanyu; Wang, Yu

    2014-09-01

    In this article nanoscaled polyanilines (PANI) were prepared based on template-free method in the presence of dicarboxylic acid dopants (e.g. D-tartaric acid, succinic acid, maleic acid and fumaric acid). The trans-cis isomerization of butenedioic acid played an important role in the formation of nanostructures from the plane-like to nanofibers, and the PANI doped with maleic acid (MA) had larger diameter, higher crystallinity and conductivity than PANI doped with fumaric acid (FA).

  18. Fast and selective determination of total protein in milk powder via titration of moving reaction boundary electrophoresis.

    PubMed

    Guo, Cheng-ye; Wang, Hou-yu; Liu, Xiao-ping; Fan, Liu-yin; Zhang, Lei; Cao, Cheng-xi

    2013-05-01

    In this paper, moving reaction boundary titration (MRBT) was developed for rapid and accurate quantification of total protein in infant milk powder, from the concept of moving reaction boundary (MRB) electrophoresis. In the method, the MRB was formed by the hydroxide ions and the acidic residues of milk proteins immobilized via cross-linked polyacrylamide gel (PAG), an acid-base indicator was used to denote the boundary motion. As a proof of concept, we chose five brands of infant milk powders to study the feasibility of MRBT method. The calibration curve of MRB velocity versus logarithmic total protein content of infant milk powder sample was established based on the visual signal of MRB motion as a function of logarithmic milk protein content. Weak influence of nonprotein nitrogen (NPN) reagents (e.g., melamine and urea) on MRBT method was observed, due to the fact that MRB was formed with hydroxide ions and the acidic residues of captured milk proteins, rather than the alkaline residues or the NPN reagents added. The total protein contents in infant milk powder samples detected via the MRBT method were in good agreement with those achieved by the classic Kjeldahl method. In addition, the developed method had much faster measuring speed compared with the Kjeldahl method.

  19. Properties of nanocellulose isolated from corncob residue using sulfuric acid, formic acid, oxidative and mechanical methods.

    PubMed

    Liu, Chao; Li, Bin; Du, Haishun; Lv, Dong; Zhang, Yuedong; Yu, Guang; Mu, Xindong; Peng, Hui

    2016-10-20

    In this work, nanocellulose was extracted from bleached corncob residue (CCR), an underutilized lignocellulose waste from furfural industry, using four different methods (i.e. sulfuric acid hydrolysis, formic acid (FA) hydrolysis, 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO)-mediated oxidation, and pulp refining, respectively). The self-assembled structure, morphology, dimension, crystallinity, chemical structure and thermal stability of prepared nanocellulose were investigated. FA hydrolysis produced longer cellulose nanocrystals (CNCs) than the one obtained by sulfuric acid hydrolysis, and resulted in high crystallinity and thermal stability due to its preferential degradation of amorphous cellulose and lignin. The cellulose nanofibrils (CNFs) with fine and individualized structure could be isolated by TEMPO-mediated oxidation. In comparison with other nanocellulose products, the intensive pulp refining led to the CNFs with the longest length and the thickest diameter. This comparative study can help to provide an insight into the utilization of CCR as a potential source for nanocellulose production. PMID:27474618

  20. Titration calculations of foot-and-mouth disease virus capsids and their stabilities as a function of pH.

    PubMed

    van Vlijmen, H W; Curry, S; Schaefer, M; Karplus, M

    1998-01-16

    Foot-and-mouth disease virus (FMDV), a non-enveloped picornavirus, is sensitive to acidic conditions. At pH values below 7 the icosahedral virus capsid, formed from 60 copies of a protomer containing four polypeptides (VP1 to 4), dissociates into 12 pentamers, releasing the viral RNA. Evidence suggests that this acid lability may assist FMDV cell entry via an endosomal pathway. Calculations of titration curves and pH-stability profiles are presented for three different strains of FMDV, O1BFS, A10(61) and A22 Iraq, and compared with experimental data for complete virions and empty capsids (which lack RNA). The finite difference Poisson-Boltzmann method was used for the calculation of electrostatic free energies with the solvent treated as a dielectric continuum. The inter-pentamer interface in the virus is formed by two protomers related by 2-fold icosahedral symmetry. As a simple model for inter-pentamer interactions, a dimer and two separate protomers were compared. The association free energy was computed by integrating the difference between the titration curves of the two species. The calculations reproduced the observed decrease in capsid stability at acidic pH but not the difference in pH sensitivities of the two type A viruses. It is shown that only residues within 15 A of the interface play a significant role in determining acid lability. For the experimentally studied pH range (5 to 7.6), histidine residues were found to dominate the pH-dependence of the stability. Two histidine residues in VP3, H142 and H145, are shown to have the greatest effect by virtue of their interactions with many polar residues across the inter-pentamer interface; the interaction of H142 with an alpha-helix in the opposite pentamer contributes only a small proportion of the destabilization energy.

  1. A visual detection of protein content based on titration of moving reaction boundary electrophoresis.

    PubMed

    Wang, Hou-Yu; Guo, Cheng-Ye; Guo, Chen-Gang; Fan, Liu-Yin; Zhang, Lei; Cao, Cheng-Xi

    2013-04-24

    A visual electrophoretic titration method was firstly developed from the concept of moving reaction boundary (MRB) for protein content analysis. In the developed method, when the voltage was applied, the hydroxide ions in the cathodic vessel moved towards the anode, and neutralized the carboxyl groups of protein immobilized via highly cross-linked polyacrylamide gel (PAG), generating a MRB between the alkali and the immobilized protein. The boundary moving velocity (V(MRB)) was as a function of protein content, and an acid-base indicator was used to denote the boundary displacement. As a proof of concept, standard model proteins and biological samples were chosen for the experiments to study the feasibility of the developed method. The experiments revealed that good linear calibration functions between V(MRB) and protein content (correlation coefficients R>0.98). The experiments further demonstrated the following merits of developed method: (1) weak influence of non-protein nitrogen additives (e.g., melamine) adulterated in protein samples, (2) good agreement with the classic Kjeldahl method (R=0.9945), (3) fast measuring speed in total protein analysis of large samples from the same source, and (4) low limit of detection (0.02-0.15 mg mL(-1) for protein content), good precision (R.S.D. of intra-day less than 1.7% and inter-day less than 2.7%), and high recoveries (105-107%).

  2. Determination of quaternary ammonium compounds by potentiometric titration with an ionic surfactant electrode: single-laboratory validation.

    PubMed

    Price, Randi; Wan, Ping

    2010-01-01

    A potentiometric titration for determining the quaternary ammonium compounds (QAC) commonly found in antimicrobial products was validated by a single laboratory. Traditionally, QACs were determined by using a biphasic (chloroform and water) manual titration procedure. Because of safety considerations regarding chloroform, as well as the subjectivity of color indicator-based manual titration determinations, an automatic potentiometric titration procedure was tested with quaternary nitrogen product formulations. By using the Metrohm Titrando system coupled with an ionic surfactant electrode and an Ag/AgCl reference electrode, titrations were performed with various QAC-containing formulation products/matrixes; a standard sodium lauryl sulfate solution was used as the titrant. Results for the products tested are sufficiently reproducible and accurate for the purpose of regulatory product enforcement. The robustness of the method was measured by varying pH levels, as well as by comparing buffered versus unbuffered titration systems. A quantitation range of 1-1000 ppm quaternary nitrogen was established. Eight commercially available antimicrobial products covering a variety of matrixes were assayed; the results obtained were comparable to those obtained by the manual titration method. Recoveries of 94 to 104% were obtained for spiked samples.

  3. Determination of quaternary ammonium compounds by potentiometric titration with an ionic surfactant electrode: single-laboratory validation.

    PubMed

    Price, Randi; Wan, Ping

    2010-01-01

    A potentiometric titration for determining the quaternary ammonium compounds (QAC) commonly found in antimicrobial products was validated by a single laboratory. Traditionally, QACs were determined by using a biphasic (chloroform and water) manual titration procedure. Because of safety considerations regarding chloroform, as well as the subjectivity of color indicator-based manual titration determinations, an automatic potentiometric titration procedure was tested with quaternary nitrogen product formulations. By using the Metrohm Titrando system coupled with an ionic surfactant electrode and an Ag/AgCl reference electrode, titrations were performed with various QAC-containing formulation products/matrixes; a standard sodium lauryl sulfate solution was used as the titrant. Results for the products tested are sufficiently reproducible and accurate for the purpose of regulatory product enforcement. The robustness of the method was measured by varying pH levels, as well as by comparing buffered versus unbuffered titration systems. A quantitation range of 1-1000 ppm quaternary nitrogen was established. Eight commercially available antimicrobial products covering a variety of matrixes were assayed; the results obtained were comparable to those obtained by the manual titration method. Recoveries of 94 to 104% were obtained for spiked samples. PMID:21140668

  4. Method for the speciation of aluminum in acidic streams

    SciTech Connect

    Miller, J.R.

    1985-01-01

    One of the impacts of acid rain falling on acid soils is the generation of high aluminum concentrations in the upper reaches of pristine mountain streams. The combination of high aluminum concentrations and lowered pH during times times of high flow is speculated by many fisheries biologists to be responsible for the decline of fish populations in these streams. In addition, this increased concern over the aquatic toxicity of aluminum has stimulated interest in the speciation of aluminum in the natural environmental. The toxicity of a metal depends greatly on its species or chemical state. The purpose of this research was to develop a practical method for speciating aluminum in those pristine streams impacted by acid rain. Aluminum has a complex chemistry involving interaction with both inorganic and organic constituents. Laboratory experiments using a chelating resin, Chelex-100, with synthetic solutions were used as the basis for speciating aluminum. The speciation procedure was applied to unfiltered samples collected from Linn Run Creek, a mildly acidic mountain stream in southwestern Pennsylvania. The results show that in upstream samples, where pH was low, rapidly exchangeable aluminum species dominated, while in the downstream samples, where pH was higher, the moderately fast exchangeable, slowly exchangeable, and inert aluminum species dominated. The laboratory experiments suggest the decline of the rapidly exchangeable fraction to be due to complexation of aluminum with humic substances. In addition, particulate aluminum was determined determined during all speciation assays.

  5. Determination of stability constants of stannous fluoride complexes by potentiostatic titration.

    PubMed

    Nelson, K G; Amin, K N

    1975-02-01

    The stability constants for stannous fluoride complexes were determined by potentiostatic titration. The method involves incremental additions of fluoride wherein each addition is followed by titration with stannous such that there is no change in the electromotive force developed between the fluoride ion and the reference electrodes. The values obtained were beta1 equals 4 times 10-3, b2 equals 1.1 times 10-7, and b3 equals 1 times 10-9. The results of this work suggest that the potentiostatic method wound be useful for determining stability constants in complexation systems involving an ion for which a specific on electrode is available.

  6. Determination of infectious bursal disease virus titration and neutralization endpoints using fluorogenic staining.

    PubMed

    Bayyari, G R; Skeeles, J K; Story, J D; Slavik, M F

    1991-01-01

    An automated method for determining infectious bursal disease virus (IBDV) titration and neutralization endpoints is described. The method employs the fluorogenic ester carboxyfluorescein-diacetate (CFDA) to stain cell monolayers in 96-well plates and a fluorescence-concentration analyzer. Titration results are compared with immunofluorescence and plaque assay titers. Virus-neutralization endpoint determination is objective, and the endpoints of replicate tests were equivalent or within one dilution of variability. Tests can be automatically screened as any percentage of a positive control or any standard deviation from a negative control.

  7. [Comparison of 4 technics for serologic titration of antibodies against rabies virus in dogs].

    PubMed

    Blancou, J; Aubert, M F; Cain, E

    1983-10-01

    Four different serological techniques were used for the determination of antibody levels against rabies virus in 55 vaccinated street dogs: mouse neutralization test (reference), rapid fluorescent focus inhibition test, plaque reduction test and immunoenzymatic test (protein A). The results obtained with each of the last three methods were compared with those obtained with the reference test: correlation coefficients were, respectively, 0.810, 0.812 and 0.682, each being significantly correlated with the others. Therefore the three techniques may each be recommended as an alternative to the mouse neutralization test for routine titrations; the immunoenzymatic method, which titrates not only neutralizing antibodies being, nevertheless, better used as a screening test.

  8. Determination of Poly(Diallyldimethylammonium Chloride) via Spectrophotometric Titration with Coomassie Brilliant Blue G Dye.

    PubMed

    Shyichuk, Alexander; Ziółkowska, Dorota; Lamkiewicz, Jan

    2016-02-01

    Poly(diallyldimethylammonium chloride) (PDDA) is used as organic coagulant in water purification and auxiliary agent in paper production. Determination of PDDA in aqueous solutions is carried out usually by means of polyelectrolyte titration using streaming current detector or o-Toluidine Blue dye for endpoint recognition. The proposed method of PDDA assay takes advantage of Coomassie Brilliant Blue G dye having strong affinity to cationic macromolecules. Titration of PDDA solution with the dye is carried out measuring absorbance at 585 nm. Inflection points of the obtained zigzag photometric curves have good repeatability provided that titration rate is low enough to attain endpoint for 1 hour or more. The method assures rectilinear calibration (R² = 0.999 5) in the broad range of PDDA concentrations from 0.3 to 9.6 mg · dm⁻³.

  9. Purity of potassium hydrogen phthalate, determination with precision coulometric and volumetric titration--a comparison.

    PubMed

    Recknagel, Sebastian; Breitenbach, Martin; Pautz, Joachim; Lück, Detlef

    2007-09-19

    The mass fraction of potassium hydrogen phthalate (KHP) from a specific batch was certified as an acidimetric standard. Two different analytical methods on a metrological level were used to carry out certification analysis: precision constant current coulometric and volumetric titration with NaOH. It could be shown that with a commercial automatic titration system in combination with a reliable software for the end-point detection it is possible to produce equivalent results with the same accuracy in comparison to a definite method handled by a fundamental apparatus for traceable precision coulometry. Prerequisite for titrations are that a high number of single measurement are applied which are calibrated with a high precision certified reference material. PMID:17870288

  10. Potentiometric/turbidometric titration of antiperspirant actives.

    PubMed

    Johnston, Clifford T; Hem, Stanley L; Guenin, Eric; Mattai, Jairajh; Afflito, John

    2003-01-01

    A titration procedure that simultaneously monitors the pH and turbidity of an antiperspirant solution during neutralization with sodium hydroxide was developed to characterize antiperspirant actives. Aluminum chloride, aluminum chlorohydrate (ACH), and aluminum zirconium glycine complex (AZG) gave distinctive pH/turbidity profiles. The activated forms of aluminum chlorohydrate (ACH') and aluminum zirconium glycine complex (AZG') produced more turbidity than the non-activated forms. On an equimolar basis, AZG' produced more turbidity than any of the antiperspirant actives tested. PMID:12715089

  11. Isolation, propagation, and titration of human immunodeficiency virus type 1 from peripheral blood of infected individuals.

    PubMed

    Schuitemaker, Hanneke; Kootstra, Neeltje A

    2005-01-01

    HIV-1 can be isolated from peripheral blood mononuclear cells and is easily propagated on primary cells in vitro. Here we describe the method for bulk isolation of the HIV-1 quasispecies and a limiting dilution virus isolation protocol by which single coexisting clones can be obtained. In addition, methods for propagation and titration of HIV-1 are provided.

  12. Electroanalytical measurements without electrolytes: conducting polymers as probes for redox titration in non-conductive organic media.

    PubMed

    Lange, Ulrich; Mirsky, Vladimir M

    2012-09-26

    Electroanalytical methods have been applied only in conducting media. An application of conducting polymers allows to overcome this limitation. If such material is in electrochemical equilibrium with dissolved redox active species, its electrical conductivity depends on the redox potential of these species. Therefore, conductometric measurements with conducting polymers can provide about the same information as classical redox electrodes. The approach was applied for redox titration. Equivalent points obtained by this titration in aqueous and organic electrolytes were identical. Then the approach was applied for determination of bromine number by redox titration in non-conducting organic phase.

  13. Hydroxyapatite-phosphonoformic acid hybrid compounds prepared by hydrothermal method

    NASA Astrophysics Data System (ADS)

    Turki, Thouraya; Othmani, Masseoud; Bantignies, Jean-Louis; Bouzouita, Khaled

    2014-01-01

    Hydroxyapatites were prepared in the presence of different amounts of phosphonoformic acid (PFA) via the hydrothermal method. The obtained powders were characterized through chemical analysis, XRD, IR, 31P MAS-NMR, TEM, and TG-TDA. The XRD showed that the PFA did not affect the apatite composition. Indeed, only a reduction of the crystallite size was noted. After grafting of PFA, the IR spectroscopy revealed the appearance of new bands belonging to HPO42- and carboxylate groups of the apatite and organic moiety, respectively. Moreover, the 31P MAS-NMR spectra exhibited a peak with a low intensity assigned to the terminal phosphonate group of the organic moiety in addition to that of the apatite. Based on these results, a reaction mechanism involving the surface hydroxyl groups (tbnd Casbnd OH) of the apatite and the carboxyl group of the acid was proposed.

  14. Titration of human coronaviruses using an immunoperoxidase assay.

    PubMed

    Lambert, Francine; Jacomy, Helene; Marceau, Gabriel; Talbot, Pierre J

    2008-01-01

    Determination of infectious viral titers is a basic and essential experimental approach for virologists. Classical plaque assays cannot be used for viruses that do not cause significant cytopathic effects, which is the case for prototype strains 229E and OC43 of human coronavirus (HCoV).Therefore, an alternative indirect immunoperoxidase assay (IPA) was developed for the detection and titration of these viruses and is described herein. Susceptible cells are inoculated with serial logarithmic dilutions of virus-containing samples in a 96-well plate format. After viral growth,viral detection by IPA yields the infectious virus titer, expressed as 'Tissue Culture Infectious Dose 50 percent' (TCID50). This represents the dilution of a virus-containing sample at which half of a series of laboratory wells contain infectious replicating virus. This technique provides are liable method for the titration of HCoV-229E and HCoV-OC43 in biological samples such as cells, tissues and fluids [corrected].

  15. Measuring the Kinetics of Molecular Association by Isothermal Titration Calorimetry.

    PubMed

    Vander Meulen, Kirk A; Horowitz, Scott; Trievel, Raymond C; Butcher, Samuel E

    2016-01-01

    The real-time power response inherent in an isothermal titration calorimetry (ITC) experiment provides an opportunity to directly analyze association kinetics, which, together with the conventional measurement of thermodynamic quantities, can provide an incredibly rich description of molecular binding in a single experiment. Here, we detail our application of this method, in which interactions occurring with relaxation times ranging from slightly below the instrument response time constant (12.5 s in this case) to as large as 600 s can be fully detailed in terms of both the thermodynamics and kinetics. In a binding titration scenario, in the most general case an injection can reveal an association rate constant (kon). Under more restrictive conditions, the instrument time constant-corrected power decay following each injection is simply an exponential decay described by a composite rate constant (kobs), from which both kon and the dissociation rate constant (koff) can be extracted. The data also support the viability of this exponential approach, for kon only, for a slightly larger set of conditions. Using a bimolecular RNA folding model and a protein-ligand interaction, we demonstrate and have internally validated this approach to experiment design, data processing, and error analysis. An updated guide to thermodynamic and kinetic regimes accessible by ITC is provided.

  16. Measuring Multivalent Binding Interactions by Isothermal Titration Calorimetry.

    PubMed

    Dam, Tarun K; Talaga, Melanie L; Fan, Ni; Brewer, Curtis F

    2016-01-01

    Multivalent glycoconjugate-protein interactions are central to many important biological processes. Isothermal titration calorimetry (ITC) can potentially reveal the molecular and thermodynamic basis of such interactions. However, calorimetric investigation of multivalency is challenging. Binding of multivalent glycoconjugates to proteins (lectins) often leads to a stoichiometry-dependent precipitation process due to noncovalent cross-linking between the reactants. Precipitation during ITC titration severely affects the quality of the baseline as well as the signals. Hence, the resulting thermodynamic data are not dependable. We have made some modifications to address this problem and successfully studied multivalent glycoconjugate binding to lectins. We have also modified the Hill plot equation to analyze high quality ITC raw data obtained from multivalent binding. As described in this chapter, ITC-driven thermodynamic parameters and Hill plot analysis of ITC raw data can provide valuable information about the molecular mechanism of multivalent lectin-glycoconjugate interactions. The methods described herein revealed (i) the importance of functional valence of multivalent glycoconjugates, (ii) that favorable entropic effects contribute to the enhanced affinities associated with multivalent binding, (iii) that with the progression of lectin binding, the microscopic affinities of the glycan epitopes of a multivalent glycoconjugate decrease (negative cooperativity), (iv) that lectin binding to multivalent glycoconjugates, especially to mucins, involves internal diffusion jumps, (bind and jump) and (v) that scaffolds of glycoconjugates influence their entropy of binding.

  17. Characterization of protein-protein interactions by isothermal titration calorimetry.

    PubMed

    Velazquez-Campoy, Adrian; Leavitt, Stephanie A; Freire, Ernesto

    2015-01-01

    The analysis of protein-protein interactions has attracted the attention of many researchers from both a fundamental point of view and a practical point of view. From a fundamental point of view, the development of an understanding of the signaling events triggered by the interaction of two or more proteins provides key information to elucidate the functioning of many cell processes. From a practical point of view, understanding protein-protein interactions at a quantitative level provides the foundation for the development of antagonists or agonists of those interactions. Isothermal Titration Calorimetry (ITC) is the only technique with the capability of measuring not only binding affinity but the enthalpic and entropic components that define affinity. Over the years, isothermal titration calorimeters have evolved in sensitivity and accuracy. Today, TA Instruments and MicroCal market instruments with the performance required to evaluate protein-protein interactions. In this methods paper, we describe general procedures to analyze heterodimeric (porcine pancreatic trypsin binding to soybean trypsin inhibitor) and homodimeric (bovine pancreatic α-chymotrypsin) protein associations by ITC.

  18. Crystallization and evaluation of hen egg-white lysozyme crystals for protein pH titration in the crystalline state.

    PubMed

    Iwai, Wakari; Yagi, Daichi; Ishikawa, Takuya; Ohnishi, Yuki; Tanaka, Ichiro; Niimura, Nobuo

    2008-05-01

    To observe the ionized status of the amino acid residues in proteins at different pH (protein pH titration in the crystalline state) by neutron diffraction, hen egg-white lysozyme was crystallized over a wide pH range (2.5-8.0). Crystallization phase diagrams at pH 2.5, 6.0 and 7.5 were determined. At pH < 4.5 the border between the metastable region and the nucleation region shifted to the left (lower precipitant concentration) in the phase diagram, and at pH > 4.5 the border shifted to the right (higher precipitant concentration). The qualities of these crystals were characterized using the Wilson plot method. The qualities of all crystals at different pH were more or less equivalent (B-factor values within 25-40). It is expected that neutron diffraction analysis of these crystals of different pH provides equivalent data in quality for discussions of protein pH titration in the crystalline state of hen egg-white lysozyme.

  19. A real-time characterization method to rapidly optimize molecular beacon signal for sensitive nucleic acids analysis.

    PubMed

    Hsieh, Albert Tsung-Hsi; Pan, Patrick J; Lee, Abraham P

    2014-05-01

    This research demonstrates an integrated microfluidic titration assay to characterize the cation concentrations in working buffer to rapidly optimize the signal-to-noise ratio (SNR) of molecular beacons (MBs). The "Microfluidic Droplet Array Titration Assay" (MiDATA) integrated the functions of sample dilution, sample loading, sample mixing, fluorescence analysis, and re-confirmation functions all together in a one-step process. It allows experimentalists to arbitrarily change sample concentration and acquire SNR measurements instantaneously. MiDATA greatly reduces sample dilution time, number of samples needed, sample consumption, and the total titration time. The maximum SNR of molecular beacons is achieved by optimizing the concentrations of the monovalent and divalent cation (i.e., Mg(2+) and K(+)) of the working buffer. MiDATA platform is able to reduce the total consumed reagents to less than 50 μL, and decrease the assay time to less than 30 min. The SNR of the designated MB is increased from 20 to 126 (i.e., enhanced the signal 630 %) using the optimal concentration of MgCl2 and KCl determined by MiDATA. This novel microfluidics-based titration method is not only useful for SNR optimization of molecular beacons but it also can be a general method for a wide range of fluorescence resonance energy transfer (FRET)-based molecular probes.

  20. Method for the isolation of citric acid and malic acid in Japanese apricot liqueur for carbon stable isotope analysis.

    PubMed

    Akamatsu, Fumikazu; Hashiguchi, Tomokazu; Hisatsune, Yuri; Oe, Takaaki; Kawao, Takafumi; Fujii, Tsutomu

    2017-02-15

    A method for detecting the undeclared addition of acidic ingredients is required to control the authenticity of Japanese apricot liqueur. We developed an analytical procedure that minimizes carbon isotope discrimination for measurement of the δ(13)C values of citric and malic acid isolated from Japanese apricot liqueur. Our results demonstrated that freeze-drying is preferable to nitrogen spray-drying, because it does not significantly affect the δ(13)C values of citric acid and results in smaller isotope discrimination for malic acid. Both 0.1% formic acid and 0.2% phosphoric acid are acceptable HPLC mobile phases for the isolation of citric and malic acid, although the δ(13)C values of malic acid exhibited relatively large variation compared with citric acid following isolation using either mobile phase. The developed procedure allows precise δ(13)C measurements of citric and malic acid isolated from Japanese apricot liqueur. PMID:27664615