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Exclusion Chromatography  

NSDL National Science Digital Library

This site contains a brief description of the separation mechanism in size exclusion chromatography. The picture helps visualize the separation, but the site features a simplified and idealized presentation useful for students new to the concept.

Kimball, John W.


Ion-Exclusion Chromatography for Analyzing Organics in Water  

NASA Technical Reports Server (NTRS)

A liquid-chromatography technique has been developed for use in the quantitative analysis of urea (and of other nonvolatile organic compounds typically found with urea) dissolved in water. The technique involves the use of a column that contains an ion-exclusion resin; heretofore, this column has been sold for use in analyzing monosaccharides and food softeners, but not for analyzing water supplies. The prior technique commonly used to analyze water for urea content has been one of high-performance liquid chromatography (HPLC), with reliance on hydrophobic interactions between analytes in a water sample and long-chain alkyl groups bonded to an HPLC column. The prior technique has proven inadequate because of a strong tendency toward co-elution of urea with other compounds. Co-elution often causes the urea and other compounds to be crowded into a narrow region of the chromatogram (see left part of figure), thereby giving rise to low chromatographic resolution and misidentification of compounds. It is possible to quantitate urea or another analyte via ultraviolet- and visible-light absorbance measurements, but in order to perform such measurements, it is necessary to dilute the sample, causing a significant loss of sensitivity. The ion-exclusion resin used in the improved technique is sulfonated polystyrene in the calcium form. Whereas the alkyl-chain column used in the prior technique separates compounds on the basis of polarity only, the ion-exclusion-resin column used in the improved technique separates compounds on the basis of both molecular size and electric charge. As a result, the degree of separation is increased: instead of being crowded together into a single chromatographic peak only about 1 to 2 minutes wide as in the prior technique, the chromatographic peaks of different compounds are now separated from each other and spread out over a range about 33 minutes wide (see right part of figure), and the urea peak can readily be distinguished from the other peaks. Although the analysis takes more time in the improved technique, this disadvantage is offset by two important advantages: Sensitivity is increased. The minimum concentration of urea that can be measured is reduced (to between 1/5 and 1/3 of that of the prior technique) because it is not necessary to dilute the sample. The separation of peaks facilitates the identification and quantitation of the various compounds. The resolution of the compounds other than urea makes it possible to identify those compounds by use of mass spectrometry.

Sauer, Richard; Rutz, Jeffrey A.; Schultz, John R.



Determination of Carboxylic Acids from Plant Root Exudates by Ion Exclusion Chromatography with ESI-MS  

Microsoft Academic Search

Ion-exclusion chromatography (IEC) coupled with electrospray ionization quadrupole mass spectrometry (ESI-MS) has been used\\u000a for the analysis of carboxylic acids. The use of ESI-MS provides increased specificity and sensitivity compared to existing\\u000a detection methods. This paper applies IEC-ESI-MS to the analysis of carboxylic acids in commonly found root exudates and shows\\u000a that the separation of nine carboxylic acids (pyruvic, oxalic,

Zuliang Chen; Kwon-Rae Kim; Gary Owens; Ravendra Naidu



Separation of Aliphatic and Aromatic Carboxylic Acids by Conventional and Ultra High Performance Ion Exclusion Chromatography  

PubMed Central

An ion exclusion chromatography (IELC) comparison between a conventional ion exchange column and an ultra-high performance liquid chromatography (UHPLC) dynamically surfactant modified C18 column for the separation of an aliphatic carboxylic acid and two aromatic carboxylic acids is presented. Professional software is used to optimize the conventional IELC separation conditions for acetylsalicylic acid and the hydrolysis products: salicylic acid and acetic acid. Four different variables are simultaneously optimized including H2SO4 concentration, pH, flow rate, and sample injection volume. Thirty different runs are suggested by the software. The resolutions and the time of each run are calculated and feed back to the software to predict the optimum conditions. Derringer’s desirability functions are used to evaluate the test conditions and those with the highest desirability value are utilized to separate acetylsalicylic acid, salicylic acid, and acetic acid. These conditions include using a 0.35 mM H2SO4 (pH 3.93) eluent at a flow rate of 1 mL min-1 and an injection volume of 72 ?L. To decrease the run time and improve the performance, a UHPLC C18 column is used after dynamic modification with sodium dodecyl sulfate. Using pure water as a mobile phase, a shorter analysis time and better resolution are achieved. In addition, the elution order is different from the IELC method which indicates the contribution of the reversed-phase mode to the separation mechanism. PMID:24285874

Mansour, Fotouh R.; Kirkpatrick, Christine L.; Danielson, Neil D.



Ion chromatography  

Microsoft Academic Search

Ion chromatography makes it possible to separate and measure low concentrations of up to 8 or 10 different anions in a single chromatographic run that takes only a few minutes. The method also works well for cations. Alkali metal ions, ammonium, magnesium, calcium, strontium, and a growing list of other metal cations and amine cations can also be rapidly separated

James S. Fritz



A process for separating acid-sugar mixtures using ion exclusion chromatography  

SciTech Connect

Work using a low-temperature concentrated sulfuric acid hydrolysis process to convert the cellulosic fraction of corn stover to monomeric sugars demonstrated the high conversion efficiencies possible with that process. The TVA work also confirmed the need for a cost-effective acid-sugar separation process. A preparative-scale ion-exclusion chromatography (IEC) system was designed, constructed, and tested with a variety of synthetic solutions and actual hydrolyzates. Although significant dispersion was observed initially, design changes were effective in minimizing this phenomenon. Data collected during the operation of the preparative-scale system were used in the design and construction of an IEC miniplant capable of processing larger volumes of synthetic solutions or hydrolyzates and in the design of an extraction-assisted IEC system. The data were also used to assess the viability of a continuous feed IEC system. This paper includes a discussion of the IEC process, provides overall material balances for various IEC process scenarios, and presents a discussion on process economics.

Hester, R.D.; Hartfield, S.W. [University of Southern Mississippi, Hattiesburg, MS (United States); Farina, G.E. [Tennessee Valley Authority, Muscle Shoals, AL (United States)



Separation of acid and sugar by ion exclusion chromatography. An application in the conversion of cellulose to ethanol  

SciTech Connect

The production of fuel grade alcohol by fermentation from sugars obtained by the acid hydrolysis of cellulose has been hindered by costly methods of cleansing the acid in the sugar stream. An economical and environmentally acceptable acid-sugar separation process based on ion exclusion chromatography has been developed and analyzed. This process recovers the acid for reuse in hydrolysis without producing landfill waste allowing a concentrated acid hydrolysis process to be commercially feasible.

Hartfield, S.; Hester, R. [Univ. of Southern Mississippi, Hattiesburg, MS (United States)



Analysis of low molecular mass organic acids in natural waters by ion exclusion chromatography tandem mass spectrometry  

Microsoft Academic Search

A sensitive and selective method for the analysis of aliphatic low molecular mass organic acids (LMMOAs) in natural waters is presented. The method is based on separation with ion exclusion chromatography and detection with electrospray ionization tandem mass spectrometry (LC–MS\\/MS). The extra selectivity gained by applying MS\\/MS allows for a minimum of sample preparation and the use of a sub-optimal

Dan Bylund; Sara H. Norström; Sofia A. Essén; Ulla S. Lundström



Biosynthesis of Cd-bound phytochelatins by Phaeodactylum tricornutum and their speciation by size-exclusion chromatography and ion-pair chromatography coupled to ICP-MS.  


Cd-bound phytochelatins (Cd-PCs) have been synthesised by incubation of Phaeodactylum tricornutum cell cultures with Cd and purified by size-exclusion chromatography-UV-Vis. These complexes, which were identified in previous work, have now been used as model substances to develop and optimise ion-pair chromatography (IPC) coupled to inductively coupled plasma-mass spectrometry (ICP-MS) for analysis of Cd-PCs. Subsequent analysis of samples taken from Silene vulgaris plants cultivated under heavy metal stress conditions revealed Cd signals but no Cd-PC signals. By use of isotopically enriched (116)Cd-PCs the sample preparation steps were verified to determine the stability of the analytes. We observed species transformation between Cd-PCs and other unidentified Cd complexes. Consequently, the kinetic and thermodynamic lability of Cd-PCs are decisive factors in their detection. PMID:16205894

Loreti, Valeria; Toncelli, Daniel; Morelli, Elisabetta; Scarano, Gioacchino; Bettmer, Jörg



Determination of short chain carboxylic acids in vegetable oils and fats using ion exclusion chromatography electrospray ionization mass spectrometry.  


A new method for quantification of short chain C1-C6 carboxylic acids in vegetable oils and fats by employing Liquid Chromatography Mass Spectrometry (LC-MS) has been developed. The method requires minor sample preparation and applies non-conventional Electrospray Ionization (ESI) liquid phase chemistry. Samples are first dissolved in chloroform and then extracted using water that has been spiked with stable isotope labeled internal standards that are used for signal normalization and absolute quantification of selected acids. The analytes are separated using Ion Exclusion Chromatography (IEC) and detected with Electrospray Ionization Mass Spectrometry (ESI-MS) as deprotonated molecules. Prior to ionization the eluent that contains hydrochloric acid is modified post-column to ensure good ionization efficiency of the analytes. The averaged within run precision and between run precision were generally lower than 8%. The accuracy was between 85 and 115% for most of the analytes. The Lower Limit of Quantification (LLOQ) ranged from 0.006 to 7mg/kg. It is shown that this method offers good selectivity in cases where UV detection fails to produce reliable results. PMID:25627971

Viidanoja, Jyrki



Ion Exchange Chromatography  

NSDL National Science Digital Library

This website contains an ion chromatography simulator that can be run online or can be downloaded. The simulator focuses on separations of proteins using Ion Chromatography. Also included is linked reference information, an example of a homework assignment using the simulator, and some information on protein structures.



Comparison of Various Preparation Methods for Determination of Organic Acids in Fruit Vinegars with a Simple Ion-Exclusion Liquid Chromatography  

Microsoft Academic Search

An ion-exclusion liquid chromatography with mobile phase 0.005 mol L?1 H2SO4 and step flow rate gradient (0.2 mL min?1 in the first 40 min and 0.5 mL min?1 from 41 to 60 min) was used to determine 20 organic acids simultaneously at 17 °C within 51 min. The peak resolutions (Rs)\\u000a were 0.45?3.02 and separation factors (?) were all higher than 1. Impurities in fruit vinegar executed with direct injection

Jau-Tien Lin; Shih-Chuan Liu; You-Cheng Shen; Deng-Jye Yang


Ion Chromatography: An Account of Its Conception and Early Development  

ERIC Educational Resources Information Center

The conception of ion chromatography and its development into a technique ready for commercialization is described. The pioneering development pointed the way to make ion exclusion an important member of the repertoire of IC methods.

Small, Hamish



Matrix-elimination with steam distillation for determination of short-chain fatty acids in hypersaline waters from pre-salt layer by ion-exclusion chromatography.  


A method for determination of formic, acetic, propionic and butyric acids in hypersaline waters by ion-exclusion chromatography (IEC), using steam distillation to eliminate matrix-interference, was developed. The steam distillation variables such as type of solution to collect the distillate, distillation time and volume of the 50% v/v H?SO? solution were optimized. The effect of the addition of NaCl different concentrations to the calibration standards on the carboxylic acid recovery was also investigated. Detection limits of 0.2, 0.5, 0.3 and 1.5 mg L?¹ were obtained for formic, acetic, propionic and butyric acids, respectively. Produced waters from petroleum reservoirs in the Brazilian pre-salt layer containing about 19% m/v of NaCl were analyzed. Good recoveries (99-108%) were obtained for all acids in spiked produced water samples. PMID:22226459

Ferreira, Fernanda N; Carneiro, Manuel C; Vaitsman, Delmo S; Pontes, Fernanda V M; Monteiro, Maria Inês C; Silva, Lílian Irene D da; Neto, Arnaldo Alcover



Electrophoretic, size-exclusion high-performance liquid chromatography and liquid chromatography-electrospray ionization ion trap mass spectrometric detection of hemoglobin-based oxygen carriers.  


Hemoglobin-based oxygen carriers (HBOCs) are blood substitutes based on hemoglobin of either bovine or human origin and they can potentially be misused in elite sports to improve endurance performance. Recently, three methods have been proposed in doping control analysis to allow HBOCs screening and identification by application of electrophoresis, size-exclusion chromatography coupled with HPLC and LC coupled with tandem mass spectrometry (LC/MSMS). In view of the Athens 2004 Olympic Games, modifications were introduced in order to increase the specificity of these methods. The sample preparation protocols of the electrophoretic and SEC-HPLC methods were modified with the introduction of sequential ultra filtration steps to remove all heme containing material below 100 kDa, thus leaving only HBOCs material for analysis. Furthermore, a modification of the LC/MSMS methodology was introduced to allow full scan MS-MS spectra of peptide segments arising from the tryptic digestion of bovine HBOCs. These relatively simple methodological modifications have major impact, as far as time and cost effectiveness is concerned in doping control procedures, because they provide a useful tool in order to identify which suspect samples from the initial visual screening are due to hemolysis and exclude them from further analysis. PMID:17386550

Simitsek, Phaedra Dora; Giannikopoulou, Panagiota; Katsoulas, Haralabos; Sianos, Efstathios; Tsoupras, George; Spyridaki, Maria-Helen; Georgakopoulos, Costas



Ion Pairing Chromatography  

Microsoft Academic Search

The present state of ion-pair chromatography (IPC) is reviewed. Emphasis is placed on the theoretical modeling of the retention behavior of charged, zwitterionic, and neutral solutes, since the theory provides the generally good understanding of the factors affecting the separation. Although stoichiometric models were credited with the first easy-to-understand demonstration of the retention mechanism of IPC, they were thermodynamic models,

Teresa Cecchi



Selective and simultaneous determination of phosphate and silicate ions in leaching process waters for ceramics glaze raw materials of narutal origin by ion-exclusion chromatography coupled with UV-detection after postcolumn derivatization.  


The selective and simultaneous ion-exclusion chromatography (IEC) with UV-detection on a weakly acidic cation-exchange resin column in the H+ -form (TSKgel Super IC-A/C) was developed and applied for the simultaneous determination of phosphate and silicate ions as the water quality parameters required for optimizing the water-leaching process for ceramics glaze raw materials of natural origin including feldspar, woods-ash, and straw-ash. Phosphate and silicate ions in these water-leaching process water samples were separated selectively from the coexisting anions such as sulfate, chloride, nitrate and carbonate ions, based on the ion-exclusion separation mechanism. They were detected selectively and simultaneously by a postcolumn derivatization with molybdenum-yellow using the UV-detector. Under the optimized separation and detection conditions (eluent, 0-1 mM sulfuric acid; reactant, 10 mM sodium molybdate-25 mM sulfuric acid; detector, UV at 370 nm; temperature, 45 degrees C), the linearity of calibration was in the range 0.1 - 10 ppm for both phosphate and silicate ions, and the detection limits at S/N = 3 were 2.58 ppb for silicate ions and 4.75 ppb for phosphate ions. The effectiveness of this method was demonstrated in practical applications to the water-leaching process for some ceramics glaze raw materials. PMID:16429785

Ikedo, Mikaru; Mori, Masanobu; Kurachi, Kazumasa; Hu, Wenzhi; Tanaka, Kazuhiko



A novel ion-exclusion chromatography-mass spectrometry method to measure concentrations and cycling rates of carbohydrates and amino sugars in freshwaters.  


The concentrations of free neutral carbohydrates and amino sugars were determined in freshwater samples of distinct matrix complexity, including meso-, eu- and dystrophic lakes and ponds, using high-performance ion-exclusion chromatography (HPIEC) coupled to mass spectrometry (MS). In contrast to other methods, our approach allowed the quantification of free neutral carbohydrates and amino sugars at low nM concentrations without derivatization, de-salting or pre-concentration. New sample preparation procedures were applied prior to injection employing syringe and hollow fiber filtration. Analytes were separated on a strong cation exchange resin under 100% aqueous conditions using 0.1% formic acid as a mobile phase. To minimize background noise in MS, analytes were detected in a multiple reaction monitoring scan mode with double ion filtering. Detection limits of carbohydrates and amino sugars ranged between 0.2 and 2nM at a signal-to-noise ratio >5. Error ranged between 1 and 12% at 0.5-500nM levels. Using a stable isotope dilution approach, both the utilization and recycling of glucose in Lake Zurich was observed. In contrast, N-acetyl-glucosamine was equally rapidly consumed but there was no visible de novo production. The simple and rapid sample preparation makes our protocol suitable for routine analyses of organic compounds in freshwater samples. Application of stable isotope tracers along with accurate measures of carbohydrate and amino sugar concentrations enables novel insights into the compound in situ dynamics. PMID:25242224

Hor?ák, Karel; Pernthaler, Jakob



Soil & Water Lab, Ion Chromatography Procedure Page 1 Ion Chromatography Procedure  

E-print Network

Soil & Water Lab, Ion Chromatography Procedure Page 1 Ion Chromatography Procedure Last updated: 5.29.13 A. Introduction Ion chromatography (IC) is used to separate out various ions based on their charge if a new machine part is needed. D. Stock Solution Preparation #12;Soil & Water Lab, Ion Chromatography

Walter, M.Todd


Ion Exchange and Liquid Column Chromatography.  

ERIC Educational Resources Information Center

Emphasizes recent advances in principles and methodology in ion exchange and chromatography. Two tables list representative examples for inorganic ions and organic compounds. Cites 544 references. (CS)

Walton, Harold F.



Direct determination of Si isotope ratios in natural waters and commercial Si standards by ion exclusion chromatography multicollector inductively coupled plasma mass spectrometry.  


Silicon isotope ratios in natural waters and several commercial Si standards were determined by online ion exclusion chromatography (IEC) multicollector inductively couple plasma mass spectrometry (MC-ICPMS). As recent studies have shown that mass-independent fractionation (MIF) also exists in MC-ICPMS, e.g., Nd, Ce, W, Sr, Hf, Ge, Hg, and Pb isotopes, the nature of mass bias for Si isotopes was thus investigated. MIF was observed for Si isotopes on both Neptune and Neptune plus MC-ICPMS instruments in this study. Therefore, a standard-sample bracketing (SSB) mass bias correction model, capable of correcting both mass-dependent and mass-independent bias, was employed to obtain accurate Si isotope ratio results in all samples by using NBS28 Si standard as the bracketing standard. Medium resolution was used for all measurements in order to resolve polyatomic interferences on Si isotopes. NBS28 Si standard solutions prepared in nutrient-free seawater and 0.1% NaOH matrix, respectively, were used for the method validation and subjected to the online IEC MC-ICPMS determination of Si isotope ratios. Values of -0.01 ± 0.06 and 0.00 ± 0.06 ‰ (1 SD, n = 10) and -0.01 ± 0.03 and 0.01 ± 0.06 ‰ (1 SD, n = 10) for ?(29/28)Si and ?(30/28)Si, respectively, were obtained, confirming accurate results can be obtained using the reported method for natural waters. Significant variations in Si isotope ratios from -0.72 ± 0.09 to -0.24 ± 0.03 ‰ (1 SD, n = 10) and -1.36 ± 0.11 to -0.46 ± 0.04 ‰ (1 SD, n = 10) for ?(29/28)Si and ?(30/28)Si, respectively, were found among commercial Si standards of NIST SRM3150, SCP Si, and Sigma-Aldrich Si. Values of -0.06 ± 0.07 and -0.20 ± 0.11 ‰ (1 SD, n = 10) for ?(29/28)Si and ?(30/28)Si, respectively, were obtained for the MOOS-3 seawater whereas 0.59 ± 0.11 and 1.19 ± 0.15 ‰ (1SD, n = 10) for ?(29/28)Si and ?(30/28)Si, respectively, were obtained for the SLRS-5 river water. To the best of our knowledge this is the first report of an application of online IEC MC-ICPMS for the high accuracy and precision determination of Si isotope ratios in natural waters. The reported method provides for a relatively rapid (10 min per run) and simple online technique that requires no sample pretreatment for the Si isotope ratio measurements. PMID:25162683

Yang, Lu; Zhou, Lian; Hu, Zhaochu; Gao, Shan



[Columns and separation mechanism of low pressure ion chromatography].  


Ion chromatography which appeared in 1975 has been developed rapidly in recent years. But the working pressure increased with years. In this work, the authors put forward the low pressure ion chromatography (LPIC) and successfully designed several kinds of low pressure ion chromatography with inlet pressure of 1.92 x 10(5)-2.94 x 10(5) Pa. Many cations, such as, alkali metals, NH4+, alkaline earth metals, transition metals, inorganic anions and organic acidic ions can be analyzed rapidly by the technique. The detectable limit of numerous ions reaches 10(-6) g/L. Low working pressure, high performance separation and highly sensitive detection can be realized by LPIC. Among these, the columns play an important part. In this paper, the kinds and species of columns and the analytes are presented. The separation mechanism of low pressure ion chromatography is discussed, which indudes ion exchange equilibrium, complexation equilibrium and ion exclusion reaction. Furthermore, the relation between the exchange capacity of ion exchange media and the concentration of eluent are also discussed. In the meanwhile, the choice of LPIC elution systems and separation conditions are described in this paper too. PMID:15739433

Zhang, X; Jiang, X



Using Ion Exchange Chromatography to Separate Proteins  

NSDL National Science Digital Library

This activity website, hosted by the National Health Museum Activities Exchange, explores protein purification using ion chromatography. Students will separate a positively charged lysozyme from a negatively charged albumin at a neutral pH using ion exchange chromatography. The site includes information for the teacher, instructions and follow-up questions for students, and a list of supplies.

Daugherty, Ellyn A.



Using Ion Exchange Chromatography to Separate Proteins  

NSDL National Science Digital Library

This activity website, hosted by the National Health Museum Activities Exchange, explores protein purification using ion chromatography. Students will separate a positively charged lysozyme from a negatively charged albumin at a neutral pH using ion exchange chromatography. The site includes information for the teacher, instructions and follow-up questions for students, and a list of supplies.

Ellyn A. Daugherty


Using Ion Exchange Chromatography to Separate and Quantify Complex Ions  

ERIC Educational Resources Information Center

Ion exchange chromatography is an important technique in the separation of charged species, particularly in biological, inorganic, and environmental samples. In this experiment, students are supplied with a mixture of two substitution-inert complex ions. They separate the complexes by ion exchange chromatography using a "flash"…

Johnson, Brian J.



Size Exclusion Chromatography with Low Angle Laser Light Scattering Detection  

Microsoft Academic Search

A review is given of the use of low angle laser light scattering (LALLS) detection in conjunction with size exclusion chromatography (SEC) to measure polymer molecular weight distributions without conventional SEC column calibration methods. A summary of light scattering theory is presented, and instrument configurations and principles of operation are described for two LALLS photometers. Also discussed are the overall

Robert C. Jordan




NSDL National Science Digital Library

The North Carolina Community College System BioNetwork's interactive eLearning tools (IETs) are reusable chunks of training that can be deployed in a variety of courses or training programs. IETs are designed to enhance, not replace hands-on training. Learners are able to enter a hands-on lab experience better prepared and more confident. This particular IET delves into chromatography including the basics, the three types (affinity, ion exchange, size exclusion), and the parts and functions associated with chromatography.


Recycling Size Exclusion Chromatography for the Analysis and Separation of Nanocrystalline Gold  

E-print Network

Recycling Size Exclusion Chromatography for the Analysis and Separation of Nanocrystalline Gold Ali University, MS-60 6100 Main Street, Houston, Texas 77025 Recycling size exclusion chromatography (RSEC defined sizes. This work aims to use size exclusion chromatography, or SEC, as a tool for evaluating

Richards-Kortum, Rebecca


Comparison of the carbohydrate biological response modifiers Krestin, schizophyllan and glucan phosphate by aqueous size exclusion chromatography with in-line argon-ion multi-angle laser light scattering photometry and differential viscometry detectors  

Microsoft Academic Search

A major barrier to the development, preclinical and clinical application of natural carbohydrate biological response modifiers has been the difficulty involved in accurately characterizing carbohydrate polymers with molecular masses ranging from 104 to 107 g\\/mol. Herein, we employed size exclusion chromatography with multi-angle laser light scattering and differential viscometry to compare and contrast structural properties of the biological response modifiers

Antje Müller; Henry A. Pretus; Rose B. McNamee; Ernest L. Jones; I. William Browder; David L. Williams




EPA Science Inventory

An analytical method was developed for the determination of lactic acid, formic acid, acetic acid, propionic acid, and butyric acid in environmental microcosm samples using ion-exclusion chromatography. The chromatographic behavior of various eluents was studied to determine the ...


The History of Ion Chromatography: The Engineering Perspective  

ERIC Educational Resources Information Center

The development of ion chromatography from an engineering perspective is presented. As ion chromatography became more widely accepted, researchers developed dozens of standard applications that enabled the creation of many low-end instruments.

Evans, Barton



Static and dynamic polystyrene dimensions in relation to the blob model and size-exclusion chromatography  

E-print Network

-exclusion chromatography Z. Gallot, G. Pouyet and A. Schmitt Centre de Recherches sur les Macromolécules, 6, rue, chromatographie d'exclusion stérique) pour une série de polystyrènes linéaires en solution dans un bon solvant-over spatial). En chromatographie d'exclusion stérique, nous montrons qu'il est possible d'établir plusieurs

Paris-Sud XI, Université de


Polydispersity in size-exclusion chromatography: a stochastic approach.  


We investigate the impact of polydispersity of the sample molecules on the separation process and on the efficiency of size-exclusion chromatography. Polydispersity was integrated into the molecular (stochastic) model of chromatography; the characteristic function, the band profile and the most important moments of the elution profiles were calculated for several kind of pore structures. We investigated the parameters affected by polydispersity on the separation for a number of pore shapes. Our results demonstrate that even a small distribution in the molecular size (i.e. polydispersity) can contribute substantially to the total width of the chromatographic peak. The pure effect of polydispersity can only be investigated via mathematical modeling, because its contribution to an experimental chromatogram cannot be separated from other band-broadening effects. PMID:25262029

Sepsey, Annamária; Bacskay, Ivett; Felinger, Attila



Size-exclusion chromatography system for macromolecular interaction analysis  


A low pressure, microcomputer controlled system employing high performance liquid chromatography (HPLC) allows for precise analysis of the interaction of two reversibly associating macromolecules such as proteins. Since a macromolecular complex migrates faster than its components during size-exclusion chromatography, the difference between the elution profile of a mixture of two macromolecules and the summation of the elution profiles of the two components provides a quantifiable indication of the degree of molecular interaction. This delta profile is used to qualitatively reveal the presence or absence of significant interaction or to rank the relative degree of interaction in comparing samples and, in combination with a computer simulation, is further used to quantify the magnitude of the interaction in an arrangement wherein a microcomputer is coupled to analytical instrumentation in a novel manner.

Stevens, Fred J. (Downers Grove, IL)



Size-exclusion chromatography using deuterated mobile phases.  


The effect of deuterated solvents in size-exclusion chromatography (SEC) was studied by comparing intrinsic viscosity measurements, SEC calibration curves, and column efficiency using water-soluble polymers. For aqueous SEC, the use of deuterium oxide slightly increases the SEC elution volume. To verify that adsorption onto the packing was absent, data from exclusion experiments were compared at 35 and 50 degrees C. Our results indicate that adsorption is not occurring for pullulan or polyethylene glycol (PEG)/poly(ethylene oxide) (PEO); for the latter, however, the elution volume increased using both D2O and H2O, indicative of slight hydrodynamic volume contraction of PEG/PEO at higher temperatures. A moderate increase in band broadening (moderate decrease in column efficiency) was observed using D2O. Finally, the effects of chloroform versus deuterated chloroform were evaluated, but no hydrodynamic volume changes were observed. PMID:16837004

Erdner, Jeanine M; Barth, Howard G; Foley, Joe P; Payne, William G



Separation of lithium ions by chromatography  

SciTech Connect

Trioctylphosphine oxide (TOPO) forms an adduct with lithium dibenzoylmethane (DBM). The chelate is readily extractable by nonaqueous solvents, and this is the basis for the chromatographic separation of lithium from other alkali ions. A liquid chromatography column with 0.1 M DBM - 0.1 M TOPO in dodecane as the stationary phase was used. Lithium ions were retrained on the column while the remaining alkali metal ions were eluted with ammonium hydroxide. Lithium was eluted with 0.6 N hydrochloric acid. The structure of the extracted lithium species was determined by extraction equilibria studies and by the method of continuous variations.

Lee, D.A.



Determination of molecular weight of heparin by size exclusion chromatography with universal calibration  

E-print Network

Determination of molecular weight of heparin by size exclusion chromatography with universal chromatography using ``universal cali- bration.'' A universal calibration curve was constructed for Superose 12 chromatography (SEC).1 * Corresponding author. Fax: 1-317-274-6879. E-mail address: (P

Dubin, Paul D.


Hexafluoroisopropanol as size exclusion chromatography mobile phase for polyamide 6.  


The present study deals with the use of hexafluoroisopropanol (HFIP) as size exclusion chromatography (SEC) mobile phase for polyamide 6 (PA6). Contradictory conclusions relating to the use of HFIP as SEC mobile phase for polyamides are found in the literature. By using a multi-detector SEC apparatus equipped with on-line viscometer and multi-angle light scattering we have studied the chromatographic artifacts and the validity of the universal calibration (UC) in HFIP for different PA6 samples (hydrolytic and anionic, monofunctional or bifunctional activator). Appropriate SEC columns and optimized experimental conditions allow most of the chromatographic artifacts to be avoided, even in neat HFIP. The use of a salt in the mobile phase, namely 0.01 M tetraethylammonium nitrate (TEAN), slightly increases the elution volume for both PA6 and PMMA polymers. Under the right conditions, the UC substantially holds for PA6. The validity of the UC is not linked to the presence of TEAN in the mobile phase. With some PA6 samples, namely those anionically synthesized using a bifunctional activator, aggregation becomes a problem and the molar mass in neat HFIP is overestimated. Addition of TEAN prevents any aggregation of the above anionically synthesized PA6. In contrast, the use of a different salt, namely potassium trifluoroacetate, increases the extent of aggregation. PMID:15387457

Mendichi, Raniero; Russo, Saverio; Ricco, Laura; Giacometti Schieroni, Alberto



[Recent advances in capillary scale ion chromatography technology].  


Ion chromatography (IC) has been a well-established technique for the analysis of ionic samples. The aqueous solution used for IC eluent is well suited for bioanalysis in relative to common liquid chromatography. This is especially true for capillary ion chromatography (CIC) due to its advantage of small sample needed. CIC is generally divided into three categories including open tubular, packed and monolithic. In this review, the recent progress of CIC is summarized based on the development of several key components associated with packed column-based system. The development of open tubular ion chromatography is also reviewed. PMID:22799186

Yang, Bingcheng; Diao, Xuefang



?-Aminobutyrohydroxamate resins as stationary phases of chelation ion chromatography  

Microsoft Academic Search

?-Aminobutyrohydroxamate resin and its derivatives were prepared and employed as stationary phases in chelation ion chromatography for the simultaneous determination of trace metal ions in sea water and biological sample. The method consists of preconcentration of a 25ml sea water or biological sample on a ?-aminobutyrohydroxamate resin column; alkali and alkaline earth metal ions are removed from the resin with

Chuen-Ying Liu; N-Ming Lee; Jian-Lian Chen



Human Plasma Proteome Analysis by Multidimensional Chromatography Prefractionation and Linear Ion Trap Mass  

E-print Network

Human Plasma Proteome Analysis by Multidimensional Chromatography Prefractionation and Linear Ion chromatography has attracted extensive attention, but most published works have focused on the fractionation strong cation exchange chromatography and reversed-phase chromatography. The resulting 30 samples were

Tian, Weidong


Specific determination of selenoaminoacids in whole milk by 2D size-exclusion-ion-paring reversed phase high-performance liquid chromatography-inductively coupled plasma mass spectrometry (HPLC-ICP MS).  


A procedure was developed for the quantitative recovery of selenomethionine (SeMet) and selenocysteine (SeCys) from whole milk. It was based on the protein unfolding, carbamidomethylation of the aminoacid residues using iodoacetamide and proteolysis using Protease XIV. The selenoaminoacids were specifically determined by ion-paring reversed phase HPLC-ICP MS after their isolation from the post-reaction mixture by size-exclusion LC. Se(IV) present in the sample was derivatized as well and was determined along with the selenoaminoacids. The origin and identity of species were identified by the co-elution with the Se(IV), isotopically labelled selenomethionine, and with the synthetic standard of carbamidomethylated selenocysteine. The method development for SeCys was assisted by using glutathione peroxidase as the SeCys standard. SeMet, SeCys and Se(IV) were quantified by the method of standard additions. The mass balance provided a measure of the method validation. The method was applied to monitoring selenium speciation during supplementation of cows (dose-effect study) with Se-rich yeast containing feed and during milk processing. PMID:18706325

Bierla, Katarzyna; Szpunar, Joanna; Lobinski, Ryszard



Molecular Characterization of Multivalent Bioconjugates by Size-Exclusion Chromatography with Multiangle Laser Light Scattering  

E-print Network

with Multiangle Laser Light Scattering Jacob F. Pollock,, Randolph S. Ashton,§ Nikhil A. Rode, David V. Schaffer polyvalent bioactive conjugates. In this study, size-exclusion chromatography with multiangle laser light scattering was paired with refractive index detection and ultraviolet spectroscopy (SEC

Schaffer, David V.


Size exclusion chromatography-low angle laser light scattering photometry of lignite macromolecules  

Microsoft Academic Search

The determination of molecular weight distribution in humic acids derived from lignite has been successfully carried out by the application of size exclusion chromatography coupled with low angle laser light scattering photometry. Reduction of soluble derivatives with zinc in acetic anhydride-dimethylformamide was required to reduce the absorbance so that linear scattering response was obtained. A benzyl derivative and a methyl

E. S. Olson; J. W. Diehl



Concentration effect in hyaluronan analysis by size exclusion chromatography  

Microsoft Academic Search

The concentration effect in size-exclusion chromatographic analysis of hyaluronans of various relative molecular masses (RMM) has been studied. A critical concentration has been found that is negatively dependent on the hyaluronan molecular mass; the higher the biopolymer molecular mass, the smaller the injected sample where the concentration effect should be taken into account for accurate evaluation of molecular mass distribution.

E. Orviský; L. Šoltés; S. Al Assaf



Theory and practice of size exclusion chromatography for the analysis of protein aggregates.  


Size exclusion chromatography (SEC) is a historical technique widely employed for the detailed characterization of therapeutic proteins and can be considered as a reference and powerful technique for the qualitative and quantitative evaluation of aggregates. The main advantage of this approach is the mild mobile phase conditions that permit the characterization of proteins with minimal impact on the conformational structure and local environment. Despite the fact that the chromatographic behavior and peak shape are hardly predictable in SEC, some generic rules can be applied for SEC method development, which are described in this review. During recent years, some improvements were introduced to conventional SEC that will also be discussed. Of these new SEC characteristics, we discuss (i) the commercialization of shorter and narrower columns packed with reduced particle sizes allowing an improvement in the resolution and throughput; (ii) the possibility of combining SEC with various detectors, including refractive index (RI), ultraviolet (UV), multi-angle laser light scattering (MALLS) and viscometer (IV), for extensive characterization of protein samples and (iii) the possibility of hyphenating SEC with mass spectrometry (MS) detectors using an adapted mobile phase containing a small proportion of organic modifiers and ion-pairing reagents. PMID:24816223

Fekete, Szabolcs; Beck, Alain; Veuthey, Jean-Luc; Guillarme, Davy



Mineral Separation in a CELSS by Ion-exchange Chromatography  

NASA Technical Reports Server (NTRS)

Operational parameters pertinent to ion exchange chromatography separation were identified. The experiments were performed with 9 mm diameter ion exchange columns and conventional column accessories. The cation separation beds were packed with AG 50W-X2 strong acid cation exchange resin in H(+) form and 200-400 dry mesh particle size. The stripper beds used in some experiments were packed with AG 1-XB strong base cation exchange resin in OH(-) form and 200-400 dry mesh particle size.

Ballou, E. V.; Spitze, L. A.; Wong, F. W.; Wydeven, T.; Johnson, C. C.




EPA Science Inventory

A method using ion chromatography (IC) for the analysis of ferrous (Fe 2+) and ferric (Fe 3+) ions in soil extracts has been developed. This method uses an ion exchange column with detection at 520 nm after post-column derivatization. Selectivity is achieved by using an anionic...


Automated Precursor Ion Exclusion During LC-MS/MS Data Acquisition for Optimal Ion Identification  

NASA Astrophysics Data System (ADS)

Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is widely used for characterizing multiple samples of complex mixtures with similar compositions. This article addresses a data acquisition strategy for collecting a maximal number of unique, high-quality MS/MS during LC-MS/MS analysis of multiple samples. Based on the concept that a component only needs to be identified once when analyzing multiple samples with similar compositions, an automated intersample data-dependent acquisition strategy was developed. The strategy is based on precursor ion exclusion (PIE) and is implemented in MassAnalyzer in an automated fashion for Thermo Scientific (San Jose, CA, USA) mass spectrometers. In this method, MassAnalyzer submits one sample at a time to the sample queue. After data acquisition of each sample, MassAnalyzer automatically analyzes the data to generate a PIE list based on the MS/MS precursor ions, merges this list with the list generated from previous runs, adds the list to the MS method file, and submits the next sample to the queue. The PIE list contains both m/z value and time window for each precursor ion, and is generated intelligently so that if an MS/MS is insufficient for identifying the peak of interest, it will be collected again near the top of the peak in the next run. Therefore, the strategy maximizes both quality and the number of unique MS/MS. When automated PIE was used to acquire LC-MS/MS data of an antibody tryptic digest and a soy hydrolysate sample, the number of identified ions increased by 52 % and 93 %, respectively, compared with data acquired without using PIE.

Zhang, Zhongqi



Application of Ion Chromatography to the Investigation of Real-World Samples Rebecca J. Whelan,*1  

E-print Network

Application of Ion Chromatography to the Investigation of Real-World Samples Rebecca J. Whelan,*1 Experiment #4: Ion Chromatography Handout prepared by David Rakestraw, Rebecca Whelan, Theresa Hannon, and Cheol Keun Chung I. Summary of Laboratory In this laboratory we will use ion chromatography to analyze

Zare, Richard N.


Multidimensional separations of complex peptide mixtures: a combined high-performance liquid chromatography/ion mobility/  

E-print Network

chromatography/ion mobility/ time-of-flight mass spectrometry approach Stephen J. Valentine1, *, Manoj Kulchania chromatography (HPLC) has been combined with high-resolution ion mobility separations and time-of-flight mass-dimensional liquid chromatography-ion mobility separations (LC-IMS) and three-dimensional LC-IMS-MS separations

Clemmer, David E.


Polysaccharide Characterization by Aqueous Size Exclusion Chromatography and Low-Angle Laser Light Scattering  

Microsoft Academic Search

Aqueous size exclusion chromatography coupled with on-line low-angle laser light scattering (SEC\\/LALLS) is a valuable analytical tool for characterization of polysaccharides and other important biopolymers. This work reviews the fundamental size separation mechanism of polymers chromatographed via SEC, the development of SEC\\/LALLS methods for characterization of eluted polymers, and applications of this technique to determine polysaccharide physical and chemical properties.

A. Corona; J. E. Rollings



Comparative study of protein molecular weights by size-exclusion chromatography and laser-light scattering  

Microsoft Academic Search

High-performance size-exclusion chromatography (SEC) based on UV-Vis detection is a relative technique for molecular weight determination whereas procedure based on multi-angle laser light scattering (MALLS) is both rapid and absolute. The two methods using recombinant human growth hormone (rHGH) and ?-lactoglobulin samples were compared. A calibration curve for the chromatographic system was generated based on standard proteins and the data

Alexis Oliva; Mat??as Llabrés; José B Fariña



Determination of lignin by size exclusion chromatography using multi angle laser light scattering  

Microsoft Academic Search

A method was developed using high-performance size exclusion liquid chromatography (HPSEC) with multi-angle laser light scattering (MALLS), quasi-elastic light scattering (QELS), interferometric refractometry (RI) and UV detection to characterize and monitor lignin. The combination proved very effective at tracking changes in molecular conformation of lignin molecules over time; i.e. changes in molecular weight distribution, radius of gyration, and hydrodynamic radius.

Aarti V. Gidh; Stephen R. Decker; Todd B. Vinzant; Michael E. Himmel; Clint Williford



Size-exclusion chromatography in 1,1,1,3,3,3-hexafluoro-2-propanol  

Microsoft Academic Search

1,1,1,3,3,3-Hexafluoroisopropanol is re-examined as an eluent for size-exclusion chromatography (SEC) of polyesters, nylons, and other polar polymers. It is shown that anomalous SEC behavior reported in previous literature can be eliminated by adding 0.01 M tetraethylammonium nitrate to the eluent. The eluent modifier does not affect the solution viscosity or root-mean-square radii of moderately polar polymers such as polyesters and

Thomas H Mourey; Trevor G Bryan



Polystyrene-equivalent molecular weight versus true molecular weight in size-exclusion chromatography  

Microsoft Academic Search

The evaluation of the size-exclusion chromatography (SEC) concentration elution curves by means of a calibration dependence obtained in a given SEC set for a polymer different from the polymer to be analyzed results in an error in the determination of both molecular weight and molecular-weight distribution (MWD). The problem is analyzed assuming the validity of the universal-calibration concept. The differences

Miloš Netopil??k; Pavel Kratochv??l



Ion chromatography of cations using indirect photometric of fluorometric detection  

SciTech Connect

Copper(II) and cerium(III) were compared as mobile phase counter-ions with a strong cation exchange column for indirect photometric chromatography (IPC). Sample ion retention time was found to be dependent upon both counter-ion size and charge, as well as mobile phase ionic strength. Detection limits of all sample ions were found to be highly dependent on the molar absorptivity of the counter-ion at the wavelength of detection. At 254nm, a cerium(III) mobile phase provided detection limits at least 100 times lower than did a copper(II) mobile phase. The detection limit of sodium was about 4 ppb, corresponding to 3.5 pmoles, using a Ce(III) mobile phase. Cerium(III) was used as a mobile phase counter-ion with a strong cation exchange column using indirect fluorescence detection. Separation of the alkali metal ions along with the ammonium ion was achieved with baseline resolution. The detection limit of sodium was 3 ppb, corresponding to 2.6 pmoles. Separation and quantitation of sodium, ammonium, and potassium ions in diluted urine was straightforward. In contrast, using indirect UV detection, interference from the sample matrix was a problem. IPC using a cerium(III) mobile phase was used to determine sodium, potassium, magnesium, and calcium ions in milk and infant formulas. Separation was complete within 17 minutes. No interference from the sample matrix was noted. Good agreement between the IPC results and atomic absorption spectroscopy was found. Several complexes of chromium(III) were studied as mobile phase candidates for indirect photometric chromatography. A complex of tris(bipyridine)chromium(III) possessed the required spectral properties of an IPC counterion, but proved to be somewhat labile in either water or methanol but not acetonitrile.

Sherman, J.H.



Ion exchange chromatography of antibody fragments.  


Effects of pH and conductivity on the ion exchange chromatographic purification of an antigen-binding antibody fragment (Fab) of pI 8.0 were investigated. Normal sulfopropyl (SP) group modified agarose particles (SP Sepharosetrade mark Fast Flow) and dextran modified particles (SP Sepharose XL) were studied. Chromatographic measurements including adsorption isotherms and dynamic breakthrough binding capacities, were complemented with laser scanning confocal microscopy. As expected static equilibrium and dynamic binding capacities were generally reduced by increasing mobile phase conductivity (1-25 mS/cm). However at pH 4 on SP Sepharose XL, Fab dynamic binding capacity increased from 130 to 160 (mg/mL media) as mobile phase conductivity changed from 1 to 5 mS/cm. Decreasing protein net charge by increasing pH from 4 to 5 at 1.3 mS/cm caused dynamic binding capacity to increase from 130 to 180 mg/mL. Confocal scanning laser microscopy studies indicate such increases were due to faster intra-particle mass transport and hence greater utilization of the media's available binding capacity. Such results are in agreement with recent studies related to ion exchange of whole antibody molecules under similar conditions. PMID:17096387

Ljunglöf, Anders; Lacki, Karol M; Mueller, Jay; Harinarayan, Chithkala; van Reis, Robert; Fahrner, Robert; Van Alstine, James M



Determination of chloride in geological samples by ion chromatography  

USGS Publications Warehouse

Samples of silicate rocks are prepared by sodium carbonate fusion and then treated by ion chromatography. The method was tested for geological standards with chloride concentration between 0.003 and 3%. Observed chloride concentrations comparedd favorably with literature values. The relative standard deviation and detection limit for the method were 8% and 7 ppm, respectively. Up to 30 determination per 24-hour period were possible. ?? 1983.

Wilson, S.A.; Gent, C.A.



Evaluation of steric exclusion chromatography on cryogel column for the separation of serum proteins.  


Steric exclusion chromatography (SXC) is a new mode of protein chromatography, in which large proteins are retained on hydrophilic stationary phase surface due to the steric exclusion of polyethylene glycol (PEG) in the mobile phase, and thereafter the retained proteins can be eluted by reducing PEG concentration. In this work, SXC was evaluated on a polyacrylamide cryogel monolith. Microscopic observation of ?-globulin precipitates on the gel surface in SXC was reported for the first time. Due to the compact packing of protein precipitates on the stationary phase surface, the dynamic retention capacity of the cryogel monolith for ?-globulin reached 20 mg/mL bed volume, much higher than those of cryogel beds in adsorption-based chromatography. The effect of molecular weight and concentration of PEG, solution pH and salt concentration on protein retention capacity was in agreement with the earlier work on SXC. Because the cryogel monoliths with interconnected macropores (10-100 ?m) allow much easy flow-through of viscous PEG buffer, the SXC can be operated at low back pressure. Hence, the cryogel monoliths are more suitable for SXC than other monoliths of narrow pores reported previously. In the separation of bovine serum proteins, albumin was recovered in the breakthrough fraction with high purity, and globulin was over eight times concentrated in the elution pool. This work has, thus, demonstrated the rapid serum protein separation and concentration by SXC on the cryogel monolith columns. PMID:24552971

Wang, Chuan; Bai, Shu; Tao, Shi-Peng; Sun, Yan



The determination of some anions using ion chromatography and ion chromatography-graphite furnace atomic absorption spectrometry  

E-print Network

chromatography. The samples were prepared for analysis by an oxygen bomb combustion technique. NBS coal standard 1632 contained 71+14 ppm F, while 1632a contained 98+10 ppm F. These values are in agreement with values reported in the literature, as determined..., but it is not without its problems. A number of interferences have been observed. In bas1c solut1ons of low fluoride concentrations, the electrode responds not only to fluor1de ion, but also to hydrox1de ion (11). At a pH less than 5. 0, hydrogen 1ons complex fluor1...

Hillman, Daniel C



Determination of UV stabilizers in PET bottles by high performance-size exclusion chromatography.  


A Size Exclusion Chromatography-High Performance Liquid Chromatography (SECHPLC) method to determine antioxidants and UV stabilizers in PET bottles has been developed. In only a single run a synthetic mixture of the stabilizers was separated and quantified. The detection limit obtained for BHT, Tinuvin 326, Cyasorb UV 5411, and Tinuvin P was about 0.1 microgram/g and for Irgafos 168 it was 1.0 microgram. RSD values were lower than 3%. Tinuvin P was identified and quantified in PET bottle extracts. Olive oil, soybean oil and sunflower oil showed well defined separation from Tinuvin P at the same conditions of analysis. Cyclic dimers were identified in the PET extracts. PMID:8799719

Monteiro, M; Nerín, C; Reyes, F G



Separation of double-walled carbon nanotubes by size exclusion column chromatography.  


In this report we demonstrate the separation of raw carbon nanotube material into fractions of double-walled (DWCNTs) and single-walled carbon nanotubes (SWCNTs). Our method utilizes size exclusion chromatography with Sephacryl gel S-200 and yielded two distinct fractions of single- and double-walled nanotubes with average diameters of 0.93 ± 0.03 and 1.64 ± 0.15 nm, respectively. The presented technique is easily scalable and offers an alternative to traditional density gradient ultracentrifugation methods. CNT fractions were characterized by atomic force microscopy and Raman and absorption spectroscopy as well as transmission electron microscopy. PMID:24896840

Moore, Katherine E; Pfohl, Moritz; Hennrich, Frank; Chakradhanula, Venkata Sai K; Kuebel, Christian; Kappes, Manfred M; Shapter, Joe G; Krupke, Ralph; Flavel, Benjamin S




NSDL National Science Digital Library

In this activity, by the Concord Consortium's Molecular Literacy project, students "learn about the phases of molecular separation, and then experiment with paper and ion chromatography." The activity itself is a java-based interactive resource built upon the free, open source Molecular Workbench software. In addition, visitors will find an overview of the activity, assessments, and concepts and their correlation to AAAS and NSES standards.


Determination of selected anions in water by ion chromatography  

USGS Publications Warehouse

Ion chromatography is a rapid, sensitive, precise, and accurate method for the determination of major anions in rainwater and surface waters. Simultaneous analyses of a single sample for bromide, chloride, fluoride, nitrate, nitrite, orthophosphate, and sulfate require approximately 20 minutes to obtain a chromatogram. Minimum detection limits range from 0.01 mg/L for fluoride to 0.20 mg/L for chloride and sulfate. Relative standard deviations were less than 9% for all anions except nitrite in Standard Reference Water Samples. Only one reference sample contained nitrite and its concentration was near the minimum level of detection. Similar precision was found for chloride, nitrate, and sulfate at concentrations less than 5 mg/L in rainfall samples. Precision for fluoride ranged from 12 to 22%, but is attributed to the low concentrations in these samples. The other anions were not detected. To determine accuracy of results, several samples were spiked with known concentrations of fluoride, chloride, nitrate, and sulfate; recoveries ranged from 96 to 103%. Known amounts of bromide and phosphate were added, separately, to several other waters, which contained bromide or phosphate. Recovery of added bromide and phosphate ranged from approximately 95 to 104%. No recovery data were obtained for nitrite. Chloride, nitrate, nitrite, orthophosphate, and sulfate, in several samples, were also determined independently by automated colorimetric procedures. An automated ion-selective electrode method was used to determine fluoride. Results are in agreement with results obtained by ion chromatography. (USGS).

Fishman, Marvin J.; Pyen, Grace



What can in situ ion chromatography offer for Mars exploration?  


The successes of the Mars exploration program have led to our unprecedented knowledge of the geological, mineralogical, and elemental composition of the martian surface. To date, however, only one mission, the Phoenix lander, has specifically set out to determine the soluble chemistry of the martian surface. The surprising results, including the detection of perchlorate, demonstrated both the importance of performing soluble ion measurements and the need for improved instrumentation to unambiguously identify all the species present. Ion chromatography (IC) is the state-of-the-art technique for soluble ion analysis on Earth and would therefore be the ideal instrument to send to Mars. A flight IC system must necessarily be small, lightweight, low-power, and have low eluent consumption. We demonstrate here a breadboard system that addresses these issues by using capillary IC at low flow rates with an optimized eluent generator and suppressor. A mix of 12 ions known or plausible for the martian soil, including 4 (oxy)chlorine species, has been separated at flow rates ranging from 1 to 10 ?L/min, requiring as little as 200 psi at 1.0 ?L/min. This allowed the use of pneumatic displacement pumping from a pressurized aluminum eluent reservoir and the elimination of the high-pressure pump entirely (the single heaviest and most energy-intensive component). All ions could be separated and detected effectively from 0.5 to 100 ?M, even when millimolar concentrations of perchlorate were present in the same mixtures. PMID:24963874

Shelor, C Phillip; Dasgupta, Purnendu K; Aubrey, Andrew; Davila, Alfonso F; Lee, Michael C; McKay, Christopher P; Liu, Yan; Noell, Aaron C



Determination of sulphite in wines using suppressed ion chromatography.  


Suppressed ion chromatography with the use of a conductivity detector was developed for the determination of sulphite ions in wine samples. When a mixed solution of sodium carbonate, sodium bicarbonate, and acetone was used as the mobile phase, simultaneous determination of eight inorganic anions (i.e., fluoride, chloride, nitrite, nitrate, sulphite, phosphate, sulphate, and thiosulphate) was completed in approximately 25min. Linearity, reproducibility, and detection limits were determined for the proposed method. In the case of sulphite detection, a linear calibration curve with a good correlation coefficient of 0.9992 was obtained from the peak height of sulphite with a relative standard deviation (n=6) 1.48%. In addition, the detection limit of sulphite was 0.27mg/L at a signal-to-noise ratio of 3. Further, the developed method was applied for the determination of sulphite contained in several wine samples. PMID:25529696

Yoshikawa, Kenji; Uekusa, Yuki; Sakuragawa, Akio




ERIC Educational Resources Information Center

This booklet presents some activities on chromatography. Directions for preparing leaf pigment extracts using alcohol are given, and paper chromatography and thin-layer chromatography are described as modifications of the basic principles of chromatography. (KHR)

Brantley, L. Reed, Sr.; Demanche, Edna L.; Klemm, E. Barbara; Kyselka, Will; Phillips, Edwin A.; Pottenger, Francis M.; Yamamoto, Karen N.; Young, Donald B.


Multicolour Fluorescence-Detection Size-Exclusion Chromatography for Structural Genomics of Membrane Multiprotein Complexes  

PubMed Central

Many interesting and important membrane proteins are hetero-oligomeric. However, besides naturally abundant examples, the structures of relatively few such complexes are known. Partly, this is due to difficulties in expression, stoichiometric assembly, and in the evaluation of their stability prior to crystallization trials. Here we describe a new approach, which allows rapid assessment of protein complex quality, assembly and stoichiometry, simplifying the search for conditions conducive to long-term stability and crystallization. Multicolour fluorescence size-exclusion chromatography (MC-FSEC) is used to enable tracking of individual subunits through expression, solubilization and purification steps. We show how the method has been applied to the heterodimeric transporter associated with antigen processing (TAP) and demonstrate how it may be extended in order to analyse membrane multisubunit assemblies. PMID:23825631

Parcej, David; Guntrum, Renate; Schmidt, Sabine; Hinz, Andreas; Tampé, Robert



Rapid size distribution and purity analysis of gastric mucus glycoproteins by size exclusion chromatography\\/multi angle laser light scattering  

Microsoft Academic Search

The weight average molar masses and molar mass distributions of two commercial pig gastric mucins and two fresh mucin preparations were determined by the technique of size exclusion chromatography coupled on-line to multi angle laser light scattering (SEC\\/MALLS). Both commercial samples exhibited much lower molar mass averages than the freshly prepared material and contained more impurities. On the other hand,

Kornelia Jumel; Immo Fiebrig; Stephen E. Harding



Characterization of atmospheric organic matter using size-exclusion chromatography with inline organic carbon detection  

NASA Astrophysics Data System (ADS)

The atmosphere contains a substantial amount of water-soluble organic material in aerosols, clouds and fogs. Despite years of efforts, little is known on the structure, composition and properties of this organic matter with most studies focusing on individual species while the bulk of the organic matter remains poorly characterized. In this work high-performance size-exclusion chromatography coupled with inline organic carbon detection (SEC-DOC) is used to characterize organic matter in fogs, clouds and aerosols collected in Fresno (CA), Whistler (BC), Davis (CA) and Selinsgrove (PA). The molecular weight distributions showed a fractional overlap of atmospheric samples and terrestrial fulvic acids although for clouds and aerosols the smaller molecular weight (MW) material is dominant. This smaller MW material is clearly resolved. Cloud and fog samples showed a larger fraction of small molecular weight organic species compared to the water-soluble fraction of aerosols, consistent with the partitioning of small molecular weight volatile species into the atmospheric aqueous phase. There are overall little differences between different sites for a same type of sample. These results obtained by one analytical set-up were confirmed with a second size-exclusion chromatography set-up using a different column and detection system. Size distributions for the same sampling location showed little inter-event variability and water-soluble organic carbon (WSOC) samples were slightly shifted toward larger sizes compared to clouds and fogs, consistent with an important contribution of volatile species to the latter ones. Cloud and aerosol samples contributed to a significant fraction (up to 21% of dissolved organic carbon (DOC)) of the macromolecular scale material.

Wang, Youliang; Chiu, Chao-An; Westerhoff, Paul; Valsaraj, Kalliat T.; Herckes, Pierre



Peptide Orientation Affects Selectivity in Ion-Exchange Chromatography  

SciTech Connect

Here we demonstrate that separation of proteolytic peptides, having the same net charge and one basic residue, is affected by their specific orientation toward the stationary phase in ion-exchange chromatography. In electrostatic repulsion-hydrophilic interaction chromatography (ERLIC) with an anion-exchange material, the C-terminus of the peptides is, on average, oriented toward the stationary phase. In cation exchange, the average peptide orientation is the opposite. Data with synthetic peptides, serving as orientation probes, indicate that in tryptic/Lys-C peptides the C-terminal carboxyl group appears to be in a zwitterionic bond with the side chain of the C-terminal Lys/Arg residue. In effect, the side chain is then less basic than the N-terminus, accounting for the specific orientation of tryptic and Lys-C peptides. Analyses of larger sets of peptides, generated from lysates by either Lys-N, Lys-C, or trypsin, reveal that specific peptide orientation affects the ability of harged side chains, such as phosphate residues, to influence retention. Phosphorylated residues that are remote in the sequence from the binding site affect retention less than those that are closer. When a peptide contains multiple charged sites, then orientation is observed to be less rigid and retention tends to be governed by the peptide’s net charge rather than its sequence. These general observations could be of value in confirming a peptide’s identification and, in particular, phosphosite assignments in proteomics analyses. More generally, orientation accounts for the ability of chromatography to separate peptides of the same compositionbut different sequence.

Alpert, Andrew J.; Petritis, Konstantinos; Kangas, Lars J.; Smith, Richard D.; Mechtler, Karl; Mitulovic, Goran; Mohammed, Shabaz; Heck, Albert J.



Peptide Orientation Affects Selectivity in Ion-Exchange Chromatography  

SciTech Connect

Here we demonstrate that separation of proteolytic peptides, having the same net charge and one basic residue, is affected by their specific orientation toward the stationary phase in ion-exchange chromatography. In electrostatic repulsion-hydrophilic interaction chromatography (ERLIC) with an anion-exchange material, the C-terminus of the peptides is, on average, oriented toward the stationary phase. In cation exchange, the average peptide orientation is the opposite. Data with synthetic peptides, serving as orientation probes, indicate that in tryptic/ Lys-C peptides the C-terminal carboxyl group appears to be in a zwitterionic bond with the side chain of the C-terminal Lys/Arg residue. In effect, the side chain is then less basic than the N-terminus, accounting for the specific orientation of tryptic and Lys-C peptides. Analyses of larger sets of peptides, generated from lysates by either Lys-N, Lys-C, or trypsin, reveal that specific peptide orientation affects the ability of charged side chains, such as phosphate residues, to influence retention. Phosphorylated residues that are remote in the sequence from the binding site affect retention less than those that are closer. When a peptide contains multiple charged sites, then orientation is observed to be less rigid and retention tends to be governed by the peptide’s net charge rather than its sequence. These general observations could be of value in confirming a peptide’s identification and, in particular, phosphosite assignments in proteomics analyses. More generally, orientation accounts for the ability of chromatography to separate peptides of the same composition but different sequence.

Alpert, Andrew J.; Petritis, Konstantinos; Kangas, Lars J.; Smith, R. D.; Mechtler, Karl; Mitulovic, Goran; Mohammed, Shabaz; Heck, Albert J.



Postcolumn techniques: A critical perspective for ion chromatography  

SciTech Connect

Part I. General overview and systems with suppressors. Postcolumn techniques represent a particularly fast-growing area in ion chromatography (IC). The developments since 1985 are surveyed in this paper. A general overview of reagent introduction means, including membrane devices and reaction conduit designs - especially various geometrically deformed open tubular designs - is provided. Part II. Determination of metals, various ionic and ionizable species. This review discusses the direct and indirect absorptiometric, fluorometric, and electrochemical detection of metal species, including rare earth elements, alkyltin, and alkyllead compounds. Part III. New methods on the horizon. The final part of this three-part review discusses chemiluminescence detection. Photolytic methods for derivatization before electrochemical detection is suggested as an important avenue for exploration. The use of postcolumn reactors built of metal tubes, wires and powder for selective reductions, electrochemical generation of reagents, fluorescence enhancement by inclusion complex formation by cyclodextrins, the coupling of reversed-phase liquid chromatography to electron capture detectors, and post separation zone compression to sharpen otherwise broad chromatographic peaks are critically reviewed.

Dasgupta, P.K. (Texas Tech Univ., Lubbock, TX (United States))



A fluorescence-detection size-exclusion chromatography-based thermostability assay to identify membrane protein expression and crystallization conditions  

PubMed Central

SUMMARY Optimization of membrane protein stability under different solution conditions is essential for obtaining crystals that diffract to high resolution. Traditional methods that evaluate protein stability require large amounts of material, and are therefore ill-suited for medium- to high-throughput screening of membrane proteins. Here we present a rapid and efficient fluorescence-detection size-exclusion chromatography-based thermostability assay (FSEC-TS). In this method, the target protein is fused to GFP. Heated protein samples, treated with a panel of additives, are then analyzed by FSEC. FSEC-TS allows one to evaluate the thermostability of nanogram to microgram amounts of the target protein under a variety of conditions without purification. We applied this method to the Danio rerio P2X4 receptor and Caenorhabditis elegans GluCl to screen ligands, ions and lipids, including newly designed cholesterol derivatives. In the case of GluCl, the screening results were used to obtain crystals of the receptor in the presence of lipids. PMID:22884106

Hattori, Motoyuki; Hibbs, Ryan E.; Gouaux, Eric



Ultra-short columns for low-pressure ion chromatography.  


Since the development of low-pressure ion chromatography (LPIC), many inorganic cations and anions as well as organic acids could be analyzed at a low-pressure of 1.96 x 10(5)-2.94 x 10(5) Pa. And the ultra-short columns took the place of the common long chromatographic columns. Furthermore, the ultra-short columns of LPIC not only reduced the system pressure appreciably, but also achieved high sensitivity and precision. In order to assess the characteristics of the super-short columns, much analysis, i.e., the analysis of alkali metals, alkaline earth metals, transition-metals, rare-earth elements, inorganic anions and organic acids as well as the common amino acids, were conducted. Moreover, excellent results were obtained from the LPIC columns. PMID:10536836

Jiang, X; Zhang, X; Liu, M



A phosphate binding assay for sevelamer hydrochloride by ion chromatography.  


Sevelamer hydrochloride is a cross-linked polymeric amine; it is the active ingredient of Renagel capsules. Renagel is indicated for the control of hyperphosphatemia in patients with end-stage renal disease. An in vitro phosphate-binding assay is required to measure the drug's efficacy. The assay developed for this purpose involves mixing the drug (polymer) with a solution of known phosphate concentration, filtering off the polymer-phosphate complex, and quantitating the unbound phosphate concentration by ion chromatography. The binding capacity, reported as mmol of phosphate bound g of polymer(-1), is calculated from the calculated amount of bound phosphate and the weight of polymer used. The method has been validated for accuracy, precision, linearity, range, and ruggedness. PMID:10698557

Mazzeo, J R; Peters, R M; Hanus, M R; Chen, X; Norton, K A



Determination of trace level ions by high-volume direct-injection ion chromatography  

Microsoft Academic Search

A high-volume direct-injection method was developed for the purpose of trace level determinations (low to sub-?g\\/l) of anions and cations by ion chromatography. The chromatographic signal was enhanced by increasing the sample volume up to 1300 ?l with no significant loss in peak efficiency. The following parameters were optimized for minimizing noise: eluent flow, background conductance, suppressor chemistry, conductivity temperature

Edward Kaiser; John Riviello; Maria Rey; John Statler; Shawn Heberling



Ion Exclusion by Sub 2-nm Carbon Nanotube Pores  

SciTech Connect

Carbon nanotubes offer an outstanding platform for studying molecular transport at nanoscale, and have become promising materials for nanofluidics and membrane technology due to their unique combination of physical, chemical, mechanical, and electronic properties. In particular, both simulations and experiments have proved that fluid flow through carbon nanotubes of nanometer size diameter is exceptionally fast compared to what continuum hydrodynamic theories would predict when applied on this length scale, and also, compared to conventional membranes with pores of similar size, such as zeolites. For a variety of applications such as separation technology, molecular sensing, drug delivery, and biomimetics, selectivity is required together with fast flow. In particular, for water desalination, coupling the enhancement of the water flux with selective ion transport could drastically reduce the cost of brackish and seawater desalting. In this work, we study the ion selectivity of membranes made of aligned double-walled carbon nanotubes with sub-2 nm diameter. Negatively charged groups are introduced at the opening of the carbon nanotubes by oxygen plasma treatment. Reverse osmosis experiments coupled with capillary electrophoresis analysis of permeate and feed show significant anion and cation rejection. Ion exclusion declines by increasing ionic strength (concentration) of the feed and by lowering solution pH; also, the highest rejection is observed for the A{sub m}{sup Z{sub A}} C{sub n}{sup Z{sub C}} salts (A=anion, C=cation, z= valence) with the greatest Z{sub A}/Z{sub C} ratio. Our results strongly support a Donnan-type rejection mechanism, dominated by electrostatic interactions between fixed membrane charges and mobile ions, while steric and hydrodynamic effects appear to be less important. Comparison with commercial nanofiltration membranes for water softening reveals that our carbon nanotube membranes provides far superior water fluxes for similar ion rejection capabilities.

Fornasiero, F; Park, H G; Holt, J K; Stadermann, M; Grigoropoulos, C P; Noy, A; Bakajin, O



High performance size exclusion chromatography. VI. Evaluation of the weight coordinate  

SciTech Connect

A series of high performance size exclusion chromatography (HPSEC) runs was made, using polystyrene standards, for the purpose of evaluating the applicability of the weight coordinate. Use of this coordinate was found to be considerably more precise than use of the time coordinate and was equivalent in precision to use of the time coordinate in conjunction with the application of an internal standard. The method consisted of continually measuring the weight of the mobile phase emerging from the columns with the Sartorius electronic balance and transmitting the voltage output of the balance to a Wang microcomputer for processing and display on a Selectric typewriter. Application of the weight coordinate obviated the use of the internal standard and permitted the shortening of run time by about one third. Experiments were performed with standards of four different molecular weights and were conducted at temperatures of 35, 45, and 55 C. The variation in temperature provided information about the temperature coefficient of HPSEC as a function of polystyrene molecular weight in the prevailing solvent. The retention weight and temperature coefficient for the internal standard, methylene chloride, was unusually large and it was hypothesized that this substance is absorbed by the polystyrene gel of the columns.

Kohn, E.



Molecular weight distribution characterization of hydrophobe-modified hydroxyethyl cellulose by size-exclusion chromatography.  


Size-exclusion chromatography (SEC) of hydrophobe-modified hydroxyethyl cellulose (HmHEC) is challenging because polymer chains are not isolated in solution due to association of hydrophobic groups and hydrophobic interaction with column packing materials. An approach to neutralize these hydrophobic interactions was developed by adding ?-cyclodextrin (?-CD) to the aqueous eluent. SEC mass recovery, especially for the higher molecular weight chains, increased with increasing concentration of ?-CD in the eluent. A ?-CD concentration of 0.75wt% in the eluent was determined to be optimal for the HmHEC polymers studied. These conditions enabled precise determinations of apparent molecular weight distributions exhibiting less than 2% relative standard deviation in the measured weight-average molecular weight (MW) for five injections on three studied samples and showed no significant differences in MW determined on two different days. The developed technology was shown to be very robust for characterizing HmHEC having MW from 500kg/mol to 2000kg/mol, and it can be potentially applied to other hydrophobe-modified polymers. PMID:25092594

Li, Yongfu; Meunier, David M; Partain, Emmett M



Oxidative protein refolding on size exclusion chromatography at high loading concentrations: Fundamental studies and mathematical modeling.  


Size exclusion chromatography has been demonstrated as an effective method for refolding a variety of proteins. However, to date process development mainly relies on laboratory experimentation of individual factors. A robust model is essential for high-throughput process screening and optimization of systems to provide higher productivity and refolding yield. In this work, a detailed kinetic scheme of oxidative refolding of a model protein (lysozyme) has been investigated to predict the refolding results in SEC. Non-reactive native, quenched and equilibrium studies were conducted to obtain the model parameters for the species formed during refolding of denatured/reduced lysozyme. The model was tested in various operating conditions, such as: protein loading concentration, injection volume, flow rate and composition of refolding buffer with and without the use of l-arginine additive. An apparent two-state mechanism was found adequate to describe refolding of lysozyme on SEC for the operating condition tested in this work. Furthermore, using low concentration of l-arginine combined with urea as common aggregation suppressor additives showed insignificant change in kinetics of refolding of lysozyme on SEC. However, addition of l-arginine changed mass transfer properties of some of the species formed in refolding reaction which was considered in the model to accurately predict the result of refolding on SEC. PMID:25454139

Saremirad, Pegah; Wood, Jeffery A; Zhang, Yan; Ray, Ajay K



Size exclusion chromatography for analyses of fibroin in silk: optimization of sampling and separation conditions  

NASA Astrophysics Data System (ADS)

A direct goal of this paper was to improve the methods of sample preparation and separation for analyses of fibroin polypeptide with the use of size exclusion chromatography (SEC). The motivation for the study arises from our interest in natural polymers included in historic textile and paper artifacts, and is a logical response to the urgent need for developing rationale-based methods for materials conservation. The first step is to develop a reliable analytical tool which would give insight into fibroin structure and its changes caused by both natural and artificial ageing. To investigate the influence of preparation conditions, two sets of artificially aged samples were prepared (with and without NaCl in sample solution) and measured by the means of SEC with multi angle laser light scattering detector. It was shown that dialysis of fibroin dissolved in LiBr solution allows removal of the salt which destroys stacks chromatographic columns and prevents reproducible analyses. Salt rich (NaCl) water solutions of fibroin improved the quality of chromatograms.

Pawcenis, Dominika; Koperska, Monika A.; Milczarek, Jakub M.; ?ojewski, Tomasz; ?ojewska, Joanna



Thin Layer Chromatography-Ion Mobility Spectrometry (TLC-IMS).  


Ion mobility spectrometry (IMS) is a fast and sensitive analytical method which operates at the atmospheric pressure. To enhance the capability of IMS for the analysis of mixtures, it is often used with preseparation techniques, such as GC or HPLC. Here, we report for the first time the coupling of the thin-layer chromatography and IMS. A variety of coupling schemes were tried that included direct electrospray from the TLC strip tip, indirect electrospray from a needle connected to the TLC strip, introducing the moving solvent into the injection port, and, the simplest way, offline introduction of scratched or cut pieces of strips into the IMS injection port. In this study a special solvent tank was designed and the TLC strip was mounted horizontally where the solvent would flow down. A very small funnel right below the TLC tip collected the solvent and transferred it to a needle via a capillary tubing. Using the TLC-ESI-IMS technique, acceptable separations were achieved for two component mixtures of morphine-papaverine and acridine-papaverine. A special injection port was designed to host the pieces cut off the TLC. The method was successfully used to identify each spot on the TLC by IMS in a few seconds. PMID:25419999

Ilbeigi, Vahideh; Tabrizchi, Mahmoud



Elucidating the redox cycle of environmental phosphorus using ion chromatography.  


Historically, it was assumed that reactive, inorganic phosphorus present in pristine environments was solely in the form of orthophosphate. However, this assumption contradicts theories of biogenesis and the observed metabolic behavior of select microorganisms. This paper discusses the role of ion chromatography (IC) in elucidating the oxidation-reduction cycle of environmental phosphorus. These methods employ suppressed-IC, coupled with tandem conductivity and electrospray mass spectrometry detectors to identify and quantify phosphorus oxyanions in natural water, synthetic cosmochemical, and biological samples. These techniques have been used to detect phosphite and orthophosphate in geothermal hot springs. Hypophosphite, phosphite, and orthophosphate have been detected in synthetic schreibersite corrosion samples, and termite extract supernatant. Synthetic schreibersite corrosion samples were also analyzed for two poly-phosphorus compounds, hypophosphate and pyrophosphate, and results show these samples did not contain concentrations above the 1.3 and 2.0 ?M respective 3? limit of detection. These methods are readily adaptable to a variety of matrices, and contribute to the elucidation of the oxidation-reduction cycle of phosphorus oxyanions in the environment. In contrast to most studies, these techniques have been used to show that phosphorus actively participates in redox processes in both the biological and geological world. PMID:21859529

Pech, Herbe; Vazquez, Maria G; Van Buren, Jean; Shi, Lixin; Ivey, Michelle M; Salmassi, Tina M; Pasek, Matthew A; Foster, Krishna L



Determination of Cyanogenic Compounds in Edible Plants by Ion Chromatography  

PubMed Central

Cyanogenic glycosides are HCN-producing phytotoxins; HCN is a powerful and a rapidly acting poison. It is not difficult to find plants containing these compounds in the food supply and/or in medicinal herb collections. The objective of this study was to investigate the distribution of total cyanide in nine genera (Dolichos, Ginkgo, Hordeum, Linum, Phaseolus, Prunus, Phyllostachys, Phytolacca, and Portulaca) of edible plants and the effect of the processing on cyanide concentration. Total cyanide content was measured by ion chromatography following acid hydrolysis and distillation. Kernels of Prunus genus are used medicinally, but they possess the highest level of total cyanide of up to 2259.81 CN?/g dry weight. Trace amounts of cyanogenic compounds were detected in foodstuffs such as mungbeans and bamboo shoots. Currently, except for the WHO guideline for cassava, there is no global standard for the allowed amount of cyanogenic compounds in foodstuffs. However, our data emphasize the need for the guidelines if plants containing cyanogenic glycosidesare to be developed as dietary supplements. PMID:24278641

Cho, Hye-Jeon; Do, Byung-Kyung; Shim, Soon-Mi; Lee, Dong-Ha; Nah, Ahn-Hee; Choi, Youn-Ju; Lee, Sook-Yeon



Molecular weight determination of lignosulfonates by size-exclusion chromatography and multi-angle laser light scattering  

Microsoft Academic Search

A lignosulfonate sample was fractionated according to the solubility in ethanol–water. The fractions were analysed by aqueous size exclusion chromatography (SEC) combined with in-line multi-angle laser light scattering (MALLS), and by static MALLS. Satisfactory SEC results were obtained with aqueous phosphate buffer containing DMSO and SDS. The refractive index increment (dn\\/dc) varied from 0.186 to 0.205 ml\\/g, depending on Mw

Guro Elise Fredheim; Svein Magne Braaten; Bjørn E Christensen



Simultaneous determination of protein aggregation, degradation, and absolute molecular weight by size exclusion chromatography–multiangle laser light scattering  

Microsoft Academic Search

The feasibility of size exclusion chromotography (SEC)–multiangle laser-light scattering as a technique to investigate aggregation and degradation of glycosylated and nonglycosylated proteins, and antibodies under various conditions such as addition of detergent, changes in pH, and variation of protein concentration and heat stress temperature was examined. Separation of proteins and their aggregates was performed using SEC–high-performance liquid chromatography. Detection of

Hongping Ye



Characterization of peroxidase:anti-peroxidase immune complexes by capillary zone electrophoresis and high-performance size-exclusion chromatography  

Microsoft Academic Search

Determination of the molecular constituents of commercial peroxidase:anti-peroxidase (PAP) preparations is necessary for the proper interpretation of PAP applications based on competitive binding assay. Capillary zone electrophoresis with field 300 V\\/cm, 40 cm capillary length (20 cm effective length), and high-performance size exclusion chromatography equipped with Superose12 HR10\\/30 column revealed that a PAP preparation used for Fc? receptor studies contained

Xi Y Mu; JianMing Lei; Chen Xu; Fred Regnier; Regina Kreisle



Analysis of a MIL-L-27502 lubricant from a gas-turbine engine test by size-exclusion chromatography  

NASA Technical Reports Server (NTRS)

Size exclusion chromatography was used to determine the chemical degradation of MIL-L-27502 oil samples from a gas turbine engine test run at a bulk oil temperature of 216 C. Results revealed a progressive loss of primary ester and additive depletion and the formation of higher molecular weight products with time. The high molecular weight products absorbed strongly in the ultraviolet indicating the presence of chromophoric groups.

Jones, W. R., Jr.; Morales, W.




NSDL National Science Digital Library

This site provides fundamental background information about chromatography, including plate theory, rate theory, the mechanisms of separations, and qualitative and quantitative aspects of chromatography. The format is a series of PowerPoint-like presentations available in PDF format.

Hardy, James K.


Ni-Cr-P plating bath time dependent characterization by ion chromatography  

E-print Network

Ni-Cr-P PLATING BATH TIME DEPENDENT CHARACTERIZATION BY ION CHROMATOGRAPHY A Thesis by RALPH EDWARD FAXEL, JR. Submitted to the Office of Graduate Studies of Texas ARM University in partial fulfillment of the requirement for the degree... of MASTER OF SCIENCE May 1990 Major Subject: Chemical Enginrering Ni-Cr-P PLATING BATH TIME DEPENDENT CHARACTERIZATION BY ION CHROMATOGRAPHY A Thesis RALPH EDWARD FAXEL, JR. Approved as to style and content by: Ralph E. White (Chair of Committee...

Faxel, Ralph Edward



Size exclusion chromatography for the unambiguous detection of aliphatics in fractions from petroleum vacuum residues, coal liquids, and standard materials, in the presence of aromatics  

Microsoft Academic Search

A method has been developed using size exclusion chromatography (SEC) in heptane eluent that can detect aliphatics unambiguously without fractionation to remove aromatics. Spherical molecules such as colloidal silicas elute at the exclusion limit, while alkanes up to Cââ elute through the porosity of the column. Detection of aliphatics was defined by use of an evaporative light scattering (ELS) detector

Eiman M. Al-Muhareb; Fatma Karaca; Trevor J. Morgan; Alan A. Herod; Ian D. Bull; Rafael Kandiyoti



Single-step isolation of extracellular vesicles by size-exclusion chromatography  

PubMed Central

Background Isolation of extracellular vesicles from plasma is a challenge due to the presence of proteins and lipoproteins. Isolation of vesicles using differential centrifugation or density-gradient ultracentrifugation results in co-isolation of contaminants such as protein aggregates and incomplete separation of vesicles from lipoproteins, respectively. Aim To develop a single-step protocol to isolate vesicles from human body fluids. Methods Platelet-free supernatant, derived from platelet concentrates, was loaded on a sepharose CL-2B column to perform size-exclusion chromatography (SEC; n=3). Fractions were collected and analysed by nanoparticle tracking analysis, resistive pulse sensing, flow cytometry and transmission electron microscopy. The concentrations of high-density lipoprotein cholesterol (HDL) and protein were measured in each fraction. Results Fractions 9–12 contained the highest concentrations of particles larger than 70 nm and platelet-derived vesicles (46%±6 and 61%±2 of totals present in all collected fractions, respectively), but less than 5% of HDL and less than 1% of protein (4.8%±1 and 0.65%±0.3, respectively). HDL was present mainly in fractions 18–20 (32%±2 of total), and protein in fractions 19–21 (36%±2 of total). Compared to the starting material, recovery of platelet-derived vesicles was 43%±23 in fractions 9–12, with an 8-fold and 70-fold enrichment compared to HDL and protein. Conclusions SEC efficiently isolates extracellular vesicles with a diameter larger than 70 nm from platelet-free supernatant of platelet concentrates. Application SEC will improve studies on the dimensional, structural and functional properties of extracellular vesicles. PMID:25279113

Böing, Anita N.; van der Pol, Edwin; Grootemaat, Anita E.; Coumans, Frank A. W.; Sturk, Auguste; Nieuwland, Rienk




EPA Science Inventory

Four collection protocols for the heart-cutting procedure in ion chromatography were compared statistically to determine the best method for use in the analysis of trace ions in water. In the comparison, the protocols were applied in the analysis of 0.1 to 0.4-ppm AS(V) added to ...


Ion-exchange chromatography separation applied to mineral recycle in closed systems  

NASA Technical Reports Server (NTRS)

As part of the controlled ecological life support system (CELSS) program, a study is being made of mineral separation on ion-exchange columns. The purpose of the mineral separation step is to allow minerals to be recycled from the oxidized waste products of plants, man, and animals for hydroponic food production. In the CELSS application, relatively large quantities of minerals in a broad concentration range must be recovered by the desired system, rather than the trace quantities and very low concentrations treated in analytical applications of ion-exchange chromatography. Experiments have been carried out to assess the parameters pertinent to the scale-up of ion-exchange chromatography and to determine feasibility. Preliminary conclusions are that the column scale-up is in a reasonable size range for the CELSS application. The recycling of a suitable eluent, however, remains a major challenge to the suitability of using ion exchange chromatography in closed systems.

Ballou, E.; Spitze, L. A.; Wong, F. W.; Wydeven, T.; Johnson, C. C.




NSDL National Science Digital Library

This lab activity from the Biotechnology Alliance for Suncoast Biology Educators covers background information on the basic types of chromatography and has the student separate pigments extracted from pens and plant leaves using paper chromatography. The lesson includes the materials needed and the demonstration procedures.

Keirle, Matt


Early milestones in the development of ion-exchange chromatography: a personal account.  


Ion chromatography as we know it today was built on a foundation of knowledge accumulated over a period of many years. Here, we review some of the outstanding earlier achievements in ion-exchange chromatography. Beginning about 1947. Spedding and Powell at Iowa State published a series of papers describing practical methods for preparative separation of the rare earths by displacement ion-exchange chromatography. The same group then demonstrated the ion-exchange separation of 14N and 15N isotopes in ammonia. Beginning in the 1950s. Kraus and Nelson at Oak Ridge published numerous analytical methods for metal ions based on separation of their chloride, fluoride, nitrate or sulfate complexes by anion chromatography. In the period from about 1960 to 1980 many clever chromatographic methods for metal ion separations were reported by researchers throughout the world and automatic in-line detection was gradually introduced. A truly innovative method by Small, Stevens and Bauman at Dow Chemical Co. marked the birth of modern ion chromatography. Anions, as well as cations, could now be separated quickly and conveniently by a system of suppressed conductivity detection. A method for anion chromatography with non-suppressed conductivity detection was published by Gjerde et al. in 1979. This was followed by a similar method for cation chromatography in 1980. Ion chromatography as we know it today did not just happen. It was built on a solid foundation of knowledge that has accumulated over a period of many years. Revisiting the older ion-exchange chromatography serves not only to pay tribute to some remarkable accomplishments, but it can also be a learning experience. Trends and ideas in science tend to run in repeating cycles. Thus, an awareness of older work may provide inspiration for new research using improved contemporary technology. Selection of milestones is a rather personal matter. I chose to write about subjects of which I came to have a firsthand knowledge during my career. The topics selected are in roughly chronological order and cover the period from about 1945 to 1980. An effort has been made to explain the chemical principles as well as to recount the major accomplishments of the various research projects. PMID:15250395

Fritz, James S



Size-exclusion chromatography of large molecules from coal liquids, petroleum residues, soots, biomass tars and humic substances.  


Size-exclusion chromatography (SEC) using 1-methyl-2-pyrrolidinone (NMP) as eluent has been calibrated using various standard polymers and model compounds and applied to the analysis of extracts of coal, petroleum and kerogens, to petroleum vacuum residues, soots, biomass tars and humic substances. Three separate columns of different molecular mass (MM) ranges were used, with detection by UV absorption; an evaporative light scattering detector was used for samples with no UV absorption. Fractionation was useful to separate signal from the less abundant high-mass material, which was normally masked by the strong signal from the more abundant low-mass material in the absence of fractionation. Fractionation methods used to isolate high-mass materials before SEC analysis included planar chromatography, column chromatography and solvent solubility. The apparently large molecules were concentrated into the fractions not soluble in common solvents and were relatively immobile in planar chromatography. All samples and fractions contained some material excluded from the column porosity. Evidence from other techniques suggests that the excluded material is of different structures from that of the resolved material rather than consisting of aggregates of small molecules. We speculate that the excluded material may elute early because the structures of this material are three-dimensional rather than planar or near planar. PMID:12834988

Herod, Alan A; Zhuo, Yuqun; Kandiyoti, Rafael




EPA Science Inventory

Selenate, selenite, and arsenate ions were separated from the major anions chloride, nitrate, and sulfate in drinking water, surface water, and groundwater sources by collecting a selected portion of the ion chromatogram, after suppression, on a concentrator column and reinjectin...


Application of ion chromatography to the study of hydrolysis of some halogenated hydrocarbons at ambient temperatures  

NASA Technical Reports Server (NTRS)

The application of ion chromatography to the study of very slow rates of hydrolysis of some halogenated hydrocarbons was investigated. The halide concentrations in the aqueous phase of mixtures of a carbonate buffer (pH = 10.3) and either chloroform (CHC13) or fluorotrichloromethane (CFC13) after aging for various lengths of time at room temperature, were determined by ion chromatography. Hydrolysis of CHC13 caused the C1(-) concentration to increase by about 1500 ppb per day. On the other hand neither the F(-) or C1(-) concentration in the CFC13 mixture increased by as much as 1 ppb per day. The magnitude of errors in the determination of halides prevented any firm conclusions regarding hydrolysis in this mixture. However, these results were used to show how ion chromatography could expedite identification of the hydrolyzing substance as well as investigations of hydrolysis mechanisms.

Otterson, D. A.




NSDL National Science Digital Library

In this activity, explore chromatography and the various colors that make up the ink in markers. Use this activity to investigate cohesion and adhesion. The online version of this activity is set up so that learners solve a mystery.

WGBH Boston



Orthogonal ion pairing reversed phase liquid chromatography purification of oligonucleotides with bulky fluorophores.  


A dual labeled oligonucleotide used as TaqMan® or 5' nuclease probe for in vitro diagnostic has been purified through orthogonal ion-pairing reversed phase chromatography, using polymeric semi-preparative and preparative PRP-1 column. We studied the mechanism of separation of oligonucleotides using ion-pairing reversed phase chromatography. We found that elution profiles of dye labeled oligonucleotides can be controlled by use of specific ion-pairing reagents. Here, we report a method for purification of an oligonucleotide containing an internally positioned rhodamine dye using two orthogonal chromatographic steps, in which the primary step resolves mostly by differences in hydrophobicity by using a weak ion-pairing reagent, and a secondary step uses a strong ion-pairing reagent for separation of length variants. Purification is demonstrated for both 1 and 15?mol scale syntheses, and amenable to further scale up for commercial lot production. PMID:24462467

Anacleto, Concordio; Ouye, Randall; Schoenbrunner, Nancy



Purge-and-trap ion chromatography for the determination of trace ammonium ion in high-salinity water samples  

Microsoft Academic Search

It has always been assumed that purge-and-trap (P&T) method is only used for the analysis of volatile organic compounds (VOCs) in aqueous samples. In this paper, a novel P&T preconcentrator has been developed for the determination of trace amounts of ammonium ion in high-salinity water samples by ion chromatography (IC). Method performance is evaluated as a function of concentration of

Po-Yen Wang; Jing-Yi Wu; Hung-Jhen Chen; Tzung-Yi Lin; Chien-Hou Wu



Dextrin characterization by high-performance anion-exchange chromatography--pulsed amperometric detection and size-exclusion chromatography--multi-angle light scattering--refractive index detection.  


Starch hydrolysis products, or dextrins, are widely used throughout the food industry for their functional properties. Dextrins are saccharide polymers linked primarily by alpha-(1 --> 4) D-glucose units and are prepared by partial hydrolysis of starch. Hydrolysis can be accomplished by the use of acid, enzymes, or by a combination of both. The hydrolysis products are typically characterized by the "dextrose equivalent" (DE), which refers to the total reducing power of all sugars present relative to glucose. While the DE gives the supplier and buyer a rough guide to the bulk properties of the material, the physiochemical properties of dextrins are dependent on the overall oligosaccharide profile. High-performance anion-exchange chromatography (HPAEC) with pulsed amperometric detection and size-exclusion chromatography (SEC) with multi-angle light-scattering and refractive index detection were used to characterize dextrins from commercial sources. HPAEC was used to acquire the oligosaccharide profile, and SEC to obtain an overall molar mass distribution. These methods in combination extended our understanding of the relationship between oligosaccharide profile, DE, and the hydrolysis process. Data from the two techniques enabled a method for estimating the DE that gave results in reasonable agreement with the accepted titration method. PMID:12830879

White, D Richard; Hudson, Patricia; Adamson, Julie T



Recent developments in electrolytic devices for ion chromatography  

Microsoft Academic Search

Recent developments in new electrolytic devices that utilize the electrolysis of water and charge-selective electromigration of ions through ion-exchange media have significantly changed the routine operation of ion chromatographic methods. Examples of these new electrolytic devices include on-line eluent generators that produce high-purity electrolyte eluents using deionized water as the carrier stream, continuously regenerated trap columns that remove ionic contaminants

Yan Liu; Kannan Srinivasan; Chris Pohl; Nebojsa Avdalovic



Determination of polyacrylamides in coal washery effluents by ultrafiltration/site-exclusion chromatography-ultraviolet detection techniques  

SciTech Connect

The use of a combined technique of ultrafiltration and aqueous size-exclusion high-performance liquid chromatography-UV detection for monitoring trace levels of residual polyacrylamide flocculants in coal washery process water is described. Flocculants of both anionic and non-ionic types in effluents are analyzed by chromatography on a TSK 5000 PW type hydrophilic and semirigid porous polymer gel with 0.05 M Na/sub 2/SO/sub 4/ in water as the mobile phase and by UV detection at 208-nm wavelength for detection. Precision studies gave a relative standard deviation of 5.8% and a precision of 2.2% at the 95% confidence level in the concentration range of 20 ppm. The lower limit of detection for the method is 1.0 Prior to chromatography, fractionation and concentration of the polyacrylamide in effluents are achieved by ultrafiltration with a hollow fiber cartridge having a nominal molecular weight cutoff of 100,000, and recoveries are determined by spiking studies. The application of the techniques for the analysis of residual flocculant in a coal washery thickener feed effluent sample is described.

Leung, R.W.M.; Pandey, R.N.; Das, B.S.



Ion-exchange sorption and preparative chromatography of biologically active materials  

SciTech Connect

This book presents information on the following topics: the problems of fine physico-chemical biotechnology; types of highly permeable network polyelectrolytes; methods for studying the permeability and porosity of network polyelectrolytes; the conformation state and flexibility of the structural elements of network polyelectrolytes; ion-exchange processes without the sorption of physiologically active substances; ion exchange, hydration, and swelling; nucleosides, nucleotides, alkaloids, sulfonamides, and miscellaneous physiologically active subtances; sharp front formation for the exchange of ions with the same valences; standard quasi-equilibrium frontal chromatography on ionites; sorption kinetics in ionites with structural heterogeneity; experimental investigations of the diffusivities of organic and physiologically active ions in ionite beads; and increasing the efficiency of low-pressure chromatography by using surface-layer and bidispersed ionites.

Samsonov, G.V.




Microsoft Academic Search

A rapid distillation method coupled with ion chromatography was developed for simultaneous determination of methylamine, ethylamine, dimethylamine, trimethylamine, propylamine, and butylamine in cosmetics. The distillation system was designed professionally to achieve rapid distillation in less than 3 min and eliminate the interferences of alkali metals, alkaline earth and organic compounds effectively. Samples were distilled using vapour stream under alkaline conditions and

Zhixiong Zhong; Gongke Li; Binghui Zhu; Zhibin Luo



Monitoring of Lactobacillus fermentation process by using ion chromatography with a series piezoelectric quartz crystal detector  

Microsoft Academic Search

A new method monitoring Lactobacillus fermentation process, which combines ion chromatography (IC) with a series piezoelectric quartz crystal (SPQC) technique, is presented in this paper. Monitoring of the fermentation process was realized by examining the rate of production of lactic acid. An automatic membrane dialyser was used for the pretreatment of the sample in on-line monitoring. A mixture of p-hydroxybenzoic

Jinzhong Zhang; Youtao Xie; Xinyu Dai; Wanzhi Wei




Technology Transfer Automated Retrieval System (TEKTRAN)

Conjugated linoleic acid (CLA; 9c,11t-18:2), present in dairy products and beef, has been reported in animals to have anticarcinogenic, growth-promoting, antiatherogenic, anti-diabetic, and lean body mass-enhancing properties. Silver ion chromatography has, over the decades, become an important met...


Application of Ion Chromatography to the Investigation of Real-World Samples  

ERIC Educational Resources Information Center

The use of ion chromatography (IC) as a means to teach important analytical concepts while giving the students a valuable opportunity to identify and investigate a real-world system of interest to them is described. A single IC apparatus can be tailored for different classes of analyses by the selection of different column-eluent combinations.

Whelan, Rebecca J.; Hannon, Theresa E.; Zare, Richard N.



Separation and simultaneous determination of four artificial sweeteners in food and beverages by ion chromatography  

Microsoft Academic Search

In this paper, the separation and determination of four artificial sweeteners (aspartame, sodium cyclamate, acesulfame-K and sodium saccharin) by ion chromatography coupled with suppressed conductivity detector is reported. The four artificial sweeteners were separated using KOH eluent generator. Due to the use of eluent generator, very low conductance background conductivity can be obtained and sensitivity of sweeteners has been greatly

Yan Zhu; Yingying Guo; Mingli Ye; Frits S. James



Analysis of anions in geological brines using ion chromatography  

SciTech Connect

Ion chromatographic procedures for the determination of the anions bromide, sulfate, nitrite, nitrate, phosphate, and iodide in brine samples have been developed and are described. The techniques have been applied to the analysis of natural brines, and geologic evaporites. Sample matrices varied over a range from 15,000 mg/L to 200,000 mg/L total halogens, nearly all of which is chloride. The analyzed anion concentrations ranged from less than 5 mg/L in the cases of nitrite, nitrate, and phosphate, to 20,000 mg/L in the case of sulfate. A technique for suppressing chloride and sulfate ions to facilitate the analysis of lower concentration anions is presented. Analysis times are typically less than 20 minutes for each procedure and the ion chromatographic results compare well with those obtained using more time consuming classical chemical analyses. 10 references, 14 figures.

Merrill, R.M.



Size Exclusion Chromatography: An Experiment for High School and Community College Chemistry and Biotechnology Laboratory Programs  

ERIC Educational Resources Information Center

A simple multiday laboratory exercise suitable for use in a high school or community college chemistry course or a biotechnology advanced placement biology course is described. In this experiment students gain experience in the use of column chromatography as a tool for the separation and characterization of biomolecules, thus expanding their…

Brunauer, Linda S.; Davis, Kathryn K.




NSDL National Science Digital Library

In this chemistry activity, learners will separate a mixture of FD&C dyes (colors certified and allowed by the US for the Food, Pharmaceutical, Cosmetics & Personal Care industry) to practice chromatography, a separation technique for mixtures. Learners will record their observations on a data table and note trends. This resource includes questions for learners and instructions for preparing the colors from M&Ms, Orange Kool-Aid, and food coloring.



Characterization of dissolved organic matter using high-performance liquid chromatography (HPLC)-size exclusion chromatography (SEC) with a multiple wavelength absorbance detector.  


High-performance liquid chromatography-size exclusion chromatography (HPLC-SEC) coupled with a multiple wavelength absorbance detector (200-445 nm) was used in this study to investigate the apparent molecular weight (AMW) distributions of dissolved organic matter (DOM). Standard DOM, namely humic acid, fulvic acid and hydrophilic acid, from the Suwannee River were tested to ascertain the performance and sensitivity of the method. In addition to four compounds groups: humic substances (Peak 1, AMW 16 kD), fulvic acids (Peak 2, AMW 11 kD), low AMW acids (Peak 3, AMW 5 kD), and low AMW neutral and amphiphilic molecules, proteins and their amino acid building blocks (Peak 4, AMW 3 kD), an new group that appears to include low AMW, 6-10 kD, humic substances was found based on investigating the spectra at various elution times. The spectroscopic parameter S(>365) (slope at wavelengths >365 nm) was determined to be a good predictor of the AMW of the DOM. The detector wavelength played an important role in evaluating the AMW distribution. For some fractions, such as the humic and low AMW non-aromatic substances, the error in measurement was ± 30% as determined by two-dimensional chromatograms detected at an artificially selected wavelength. HPLC-SEC with multiple wavelength absorbance detection was found to be a useful technique for DOM characterization. It characterized the AMW distributions of DOM more accurately and provided additional, potentially important information concerning the properties of DOM with varying AMWs. PMID:22369846

Yan, Mingquan; Korshin, Gregory; Wang, Dongsheng; Cai, Zhenxiao



Trace enrichment and separation of metal ions as dithiocarbamate complexes by liquid chromatography  

Microsoft Academic Search

A method is presented that involves the simultaneous formation of metal dithiocarbamates and on-line preconcentration and, subsequently, separation of heavy-metal ions (Cd(II), Pb(II), Hg(II), Cu(II), Co(II), Ni(II), Bi(III)) by reversed-phase liquid chromatography. A cetrimide-dithiocarbamate ion pair is loaded onto a precolumn packed with Cââ-bonded silica, and the injected metal ions react instantaneously with the dithiocarbamate to form stable complexes. These

Hubertus. Irth; G. J. de Jong; U. A. Th. Brinkman; R. W. Frei



Molecular characterization of multivalent bioconjugates by size-exclusion chromatography (SEC) with multi-angle laser light scattering (MALS)  

PubMed Central

The degree of substitution and valency of bioconjugate reaction products are often poorly judged or require multiple time- and product- consuming chemical characterization methods. These aspects become critical when analyzing and optimizing the potency of costly polyvalent bioactive conjugates. In this study, size-exclusion chromatography with multi-angle laser light scattering was paired with refractive index detection and ultraviolet spectroscopy (SEC-MALS-RI-UV) to characterize the reaction efficiency, degree of substitution, and valency of the products of conjugation of either peptides or proteins to a biopolymer scaffold, i.e., hyaluronic acid (HyA). Molecular characterization was more complete compared to estimates from a protein quantification assay, and exploitation of this method led to more accurate deduction of the molecular structures of polymer bioconjugates. Information obtained using this technique can improve macromolecular engineering design principles and better understand multivalent macromolecular interactions in biological systems. PMID:22794081

Pollock, Jacob F.; Ashton, Randolph S.; Rode, Nikhil A.; Schaffer, David V.; Healy, Kevin E.



Determination of pore size distributions in capillary-channeled polymer fiber stationary phases by inverse size-exclusion chromatography and implications for fast protein separations.  


Capillary-channeled polymer (C-CP) fibers have been utilized as liquid chromatography stationary phases, primarily for biomacromolecule separations on the analytical and preparative scales. The collinear packing of the eight-channeled C-CP fibers provides for very efficient flow, allowing operation at high linear velocity (u>100mm s(-1)) and low backpressure (<2000psi) in analytical-scale separations. To take advantage of these fluid transport properties, there must not be mass transfer limitations as would be imposed by having an appreciably porous phase, wherein solute diffusion limits the overall mass transport rates. To better understand the physical nano-/micro- structure of C-CP fibers, inverse size exclusion chromatography (iSEC) has been employed to determine the pore size distribution (PSD) within C-CP fibers. A diversity of test species (from metal ions to large proteins) was used as probes under non-retaining conditions to obtain a response curve reflecting the apparent partition coefficient (Kd) versus hydrodynamic radii (rm). A mean pore radius (rp) of 4.2nm with standard deviation (sp) of ±1.1nm was calculated by fitting the Kd versus rm data to model equations with a Gaussian pore size distribution, and a pore radius of 4.0±0.1nm was calculated based on a log-normal distribution. The derived mean pore radius is much smaller than traditional support materials, with the standard deviation showing a relatively uniform pore distribution. van Deemter plots were analyzed to provide practical confirmation of the structural implications. Large molecules (e.g., proteins) that are fully excluded from pores have no significant C-terms in the van Deemter plots whereas small molecules that can access the pore volumes display appreciable C-terms, as expected. Fitting of retention data to the Knox equation suggests that the columns operate with a characteristic particle diameter (dp) of ?53?m. PMID:24877979

Wang, Zhengxin; Marcus, R Kenneth



The Determination of Hyaluronan Molecular Weight Distribution by Means of High-Performance Size Exclusion Chromatography  

Microsoft Academic Search

The high-performance liquid chromatographic (HPLC) method relevant to the size exclusion chromatographic mode (SEC) used for the distribution analysis of high-molecular-weight hyaluronans is described. The HPLC fillings used, to which was applied a phosphate buffer (50 mM, pH 7.8) effluent, consisted of a cross-linked hydroxyethylmethacrylate derivative gel (HEMA-BIO), which is commercially available, and a highly porous aminopropyl-silica sorbent (SG-10-6000-NH2), which

E. Orviský; L. Šoltés; P. Chabr?ek; I. Novák; V. Kéry; M. Stan?íková; I. Vinš



Fractionation of cis- and trans-oleic, linoleic, and conjugated linoleic fatty acid methyl esters by silver ion high-performance liquid chromatography 1 Names are necessary to report factually on available data; however, the USDA neither guarantees nor warrants the standard of the product, and the use of the name by USDA implies no approval of the product to the exclusion of others that may also be suitable. 1  

Microsoft Academic Search

Silver ion high-performance liquid chromatography (Ag-HPLC) using two commercially available columns connected in series and a solvent of 0.5% acetonitrile in hexane (1.0 ml\\/min), was used to fractionate a mixture of conjugated 18:2 isomers (as methyl esters) and resulted in an elution order (10 ?g sample size) of: 9-trans,11-trans-18:2; 9-cis,11-trans-18:2. A more complex mixture of cis and trans 9-18:1, 9,11-18:2

R Adlof; T Lamm



Fabricating electrospun cellulose nanofibre adsorbents for ion-exchange chromatography.  


Protein separation is an integral step in biopharmaceutical manufacture with diffusion-limited packed bed chromatography remaining the default choice for industry. Rapid bind-elute separation using convective mass transfer media offers advantages in productivity by operating at high flowrates. Electrospun nanofibre adsorbents are a non-woven fibre matrix of high surface area and porosity previously investigated as a bioseparation medium. The effects of compression and bed layers, and subsequent heat treatment after electrospinning cellulose acetate nanofibres were investigated using diethylaminoethyl (DEAE) or carboxylate (COO) functionalisations. Transbed pressures were measured and compared by compression load, COO adsorbents were 30%, 70% and 90% higher than DEAE for compressions 1, 5 and 10MPa, respectively, which was attributed to the swelling effect of hydrophilic COO groups. Dynamic binding capacities (DBCs) at 10% breakthrough were measured between 2000 and 12,000CV/h (2s and 0.3s residence times) under normal binding conditions, and DBCs increased with reactant concentration from 4 to 12mgBSA/mL for DEAE and from 10 to 21mglysozyme/mL for COO adsorbents. Comparing capacities of compression loads applied after electrospinning showed that the lowest load tested, 1MPa, yielded the highest DBCs for DEAE and COO adsorbents at 20mgBSA/mL and 27mglysozyme/mL, respectively. At 1MPa, DBCs were the highest for the lowest flowrate tested but stabilised for flowrates above 2000CV/h. For compression loads of 5MPa and 10MPa, adsorbents recorded lower DBCs than 1MPa as a result of nanofibre packing and reduced surface area. Increasing the number of bed layers from 4 to 12 showed decreasing DBCs for both adsorbents. Tensile strengths were recorded to indicate the mechanical robustness of the adsorbent and be related to packing the nanofibre adsorbents in large scale configurations such as pleated cartridges. Compared with an uncompressed adsorbent, compressions of 1, 5 and 10MPa showed increases of 30%, 110% and 110%, respectively, for both functionalisations. The data presented show that capacity and mechanical strength can be balanced through compression after electrospinning and is particular to different functionalisations. This trade-off is critical to the development of nanofibre adsorbents into different packing configurations for application and scale up in bioseparation. PMID:25541092

Dods, Stewart R; Hardick, Oliver; Stevens, Bob; Bracewell, Daniel G



Fabricating electrospun cellulose nanofibre adsorbents for ion-exchange chromatography  

PubMed Central

Protein separation is an integral step in biopharmaceutical manufacture with diffusion-limited packed bed chromatography remaining the default choice for industry. Rapid bind-elute separation using convective mass transfer media offers advantages in productivity by operating at high flowrates. Electrospun nanofibre adsorbents are a non-woven fibre matrix of high surface area and porosity previously investigated as a bioseparation medium. The effects of compression and bed layers, and subsequent heat treatment after electrospinning cellulose acetate nanofibres were investigated using diethylaminoethyl (DEAE) or carboxylate (COO) functionalisations. Transbed pressures were measured and compared by compression load, COO adsorbents were 30%, 70% and 90% higher than DEAE for compressions 1, 5 and 10 MPa, respectively, which was attributed to the swelling effect of hydrophilic COO groups. Dynamic binding capacities (DBCs) at 10% breakthrough were measured between 2000 and 12,000 CV/h (2 s and 0.3 s residence times) under normal binding conditions, and DBCs increased with reactant concentration from 4 to 12 mg BSA/mL for DEAE and from 10 to 21 mg lysozyme/mL for COO adsorbents. Comparing capacities of compression loads applied after electrospinning showed that the lowest load tested, 1 MPa, yielded the highest DBCs for DEAE and COO adsorbents at 20 mg BSA/mL and 27 mg lysozyme/mL, respectively. At 1 MPa, DBCs were the highest for the lowest flowrate tested but stabilised for flowrates above 2000 CV/h. For compression loads of 5 MPa and 10 MPa, adsorbents recorded lower DBCs than 1 MPa as a result of nanofibre packing and reduced surface area. Increasing the number of bed layers from 4 to 12 showed decreasing DBCs for both adsorbents. Tensile strengths were recorded to indicate the mechanical robustness of the adsorbent and be related to packing the nanofibre adsorbents in large scale configurations such as pleated cartridges. Compared with an uncompressed adsorbent, compressions of 1, 5 and 10 MPa showed increases of 30%, 110% and 110%, respectively, for both functionalisations. The data presented show that capacity and mechanical strength can be balanced through compression after electrospinning and is particular to different functionalisations. This trade-off is critical to the development of nanofibre adsorbents into different packing configurations for application and scale up in bioseparation. PMID:25541092

Dods, Stewart R.; Hardick, Oliver; Stevens, Bob; Bracewell, Daniel G.



Detection of Alkylaminium Salts in Particulate Matter by Ion Chromatography  

NASA Astrophysics Data System (ADS)

Smog chamber experiments were conducted to determine how amines react to form particles, specifically amine salts, in the atmosphere. All of the experiments were performed in a smog chamber at University of California Riverside's College of Engineering Center for Environmental Research and Technology (CE-CERT). A Particle Into Liquid Sampler Ion Chromatograph (PILS-IC) was used to determine the concentration of the amine salts formed during the experiment. It became apparent that the amines (trimethylamine, diethylamine, and butylamine) behave differently in the presence of oxidants. The oxidants used were N2O5 and hydroxyl, both under varying levels of humidity (0 - 40%). By order of decreasing amount, the amines that produced the highest concentration of amine salts were: diethylamine, butylamine, and trimethylamine. It was also discovered that the hydroxyl radical had a higher tendency to oxidize the carbon side chain of the amine, rather than form the salt as in the case of N2O5.

Praske, E.; Lee, S. A.; Tang, X.; Cocker, D. R.; Silva, P. J.; Purvis-Roberts, K. L.



Screening of Drugs and Toxic Compounds with Liquid Chromatography-Linear Ion Trap Tandem Mass Spectrometry  

Microsoft Academic Search

Background: In clinical and forensic toxicology, general unknown screening is used to detect and identify exog- enous compounds. In this study, we aimed to develop a comprehensive general unknown screening method based on liquid chromatography coupled with a hybrid triple-quadrupole linear ion trap mass spectrometer. Methods: After solid-phase extraction, separation was performed using gradient reversed-phase chromatogra- phy. The mass spectrometer

Francois-Ludovic Sauvage; Franck Saint-marcoux; Benedicte Duretz; Didier Deporte; Gerard Lachatre; Pierre Marquet


Determination of copper and zinc in blood plasma by ion chromatography using a cobalt internal standard  

Microsoft Academic Search

Ion chromatography was used to detect levels of copper and zinc in blood plasma from renal dialysis patients on continuous ambulatory peritoneal dialysis (CAPD) and haemodialysis (HD). The developed method used cobalt as an internal standard and when combined with the standard additions method improved the overall precision of the results by between 20.3 and 6.0% and 20.8 and 5.7%

Alexis J. Holden; Robert A. Cowardb



Urinary NO2- and NO3- evaluation by an ion chromatography system.  


We describe an ion chromatography system using a Dionex AS4A-SC column with carbonate-bicarbonate buffer (1.8-1.7 mM) as eluent for the evaluation of urinary NO2- and NO3-. This chromatographic system gives an accurate measurement of NO2- and NO3- in the urine as an index of NO production in vivo, making also possible to evaluate their relative proportion and providing useful tools to investigate the NO system. PMID:9568279

Calo, L; Cantaro, S; Paleari, D; Vianello, D; Zerbo, F; Bonfante, L; Favaro, S; Antonello, A; D'Angelo, A



Protein-surface interaction maps for ion-exchange chromatography.  


In this paper, protein-surface interaction maps were generated by performing coarse-grained protein-surface calculations. This approach allowed for the rapid determination of the protein-surface interaction energies at a range of orientations and distances. Interaction maps of lysozyme indicated that there was a contiguous series of orientations corresponding to several adjacent preferred binding regions on the protein surface. Examination of these orientations provided insight into the residues involved in surface interactions, which qualitatively agreed with the retention data for single-site mutants. Interaction maps of lysozyme single-site mutants were also generated and provided significant insight into why these variants exhibited significant differences in their chromatographic behavior. This approach was also employed to study the binding behavior of CspB and related mutants. The results indicated that, in addition to describing general trends in the data, these maps provided significant insight into retention data of the single-site mutants. In particular, subtle retention trends observed with the K12 and K13 mutants were well-described using this interaction map approach. Finally, the number of interaction points with energies stronger than -2 kcal/mol was shown to be able to semi-quantitatively predict the behavior of most of the mutants. This rapid approach for calculating protein-surface interaction maps is expected to facilitate future method development for separating closely related protein variants in ion-exchange systems. PMID:21375221

Freed, Alexander S; Cramer, Steven M



Determination of alkylamines in atmospheric aerosol particles: a comparison of gas chromatography-mass spectrometry and ion chromatography approaches  

NASA Astrophysics Data System (ADS)

In recent years low molecular weight alkylamines have been recognized to play an important role in particle formation and growth in the lower atmosphere. However, major uncertainties are associated with their atmospheric processes, sources and sinks, mostly due to the lack of ambient measurements and the difficulties in accurate quantification of alkylamines at trace level. In this study, we present the evaluation and optimization of two analytical approaches, i.e., gas chromatography-mass spectrometry (GC-MS) and ion chromatography (IC), for the determination of alkylamines in aerosol particles. Alkylamines were converted to carbamates through derivatization with isobutyl chloroformate for GC-MS determination. A set of parameters affecting the analytical performances of the GC-MS approach, including reagent amount, reaction time and pH value, was evaluated and optimized. The accuracy is 84.3-99.1%, and the limits of detection obtained are 1.8-3.9 pg (or 0.02-0.04 ng m-3). For the IC approach, a solid-phase extraction (SPE) column was used to separate alkylamines from interfering cations before IC analysis. 1-2% (v/v) of acetone (or 2-4% (v/v) of acetonitrile) was added to the eluent to improve the separation of alkylamines on the IC column. The limits of detection obtained are 2.1-15.9 ng (or 0.9-6.4 ng m-3), and the accuracy is 55.1-103.4%. The lower accuracy can be attributed to evaporation losses of amines during the sample concentration procedure. Measurements of ambient aerosol particle samples collected in Hong Kong show that the GC-MS approach is superior to the IC approach for the quantification of primary and secondary alkylamines due to its lower detection limits and higher accuracy.

Huang, R.-J.; Li, W.-B.; Wang, Y.-R.; Wang, Q. Y.; Jia, W. T.; Ho, K.-F.; Cao, J. J.; Wang, G. H.; Chen, X.; Haddad, I. EI; Zhuang, Z. X.; Wang, X. R.; Prévôt, A. S. H.; O'Dowd, C. D.; Hoffmann, T.



Determination of alkyl amines in atmospheric aerosol particles: a comparison of gas chromatography-mass spectrometry and ion chromatography approaches  

NASA Astrophysics Data System (ADS)

In recent years low molecular weight alkyl amines have been recognized to play an important role in particle formation and growth in the lower atmosphere. However, major uncertainties are associated with their atmospheric processes, sources and sinks, mostly due to the lack of ambient measurements and the difficulties in accurate quantification of alkyl amines at trace level. In this study, we present the evaluation and optimization of two analytical approaches, i.e., gas chromatography-mass spectrometry (GC-MS) and ion chromatography (IC), for the determination of alkyl amines in aerosol particles. Alkyl amines were converted to carbamates through derivatization with isobutyl chloroformate for GC-MS determination. A set of parameters affecting the analytical performances of the GC-MS approach, including reagent amount, reaction time and pH value, was evaluated and optimized. The accuracy is 84.3-99.1%, and the limits of detection obtained are 1.8-3.9 pg. For the IC approach, a solid phase extraction (SPE) column was used to separate alkyl amines from interfering cations before IC analysis. 1-2% (v/v) of acetone (or 2-4% (v/v) of acetonitrile) was added to the eluent to improve the separation of alkyl amines on the IC column. The limits of detection obtained are 2.1-15.9 ng and the accuracy is 55.1-103.4%. The lower accuracy can be attributed to evaporation losses of amines during the sample concentration procedure. Measurements of ambient aerosol particle samples collected in Hong Kong show that the GC-MS approach is superior to the IC approach for the quantification of primary and secondary alkyl amines due to its lower detection limits and higher accuracy.

Huang, R.-J.; Li, W.-B.; Wang, Y.-R.; Wang, Q. Y.; Ho, K.-F.; Cao, J. J.; Wang, G. H.; Chen, X.; Haddad, I. EI; Zhuang, Z. X.; Wang, X. R.; Prévôt, A. S. H.; O'Dowd, C. D.; Hoffmann, T.



Megadalton Complexes in the Chloroplast Stroma of Arabidopsis thaliana Characterized by Size Exclusion Chromatography, Mass Spectrometry, and Hierarchical Clustering*  

PubMed Central

To characterize MDa-sized macromolecular chloroplast stroma protein assemblies and to extend coverage of the chloroplast stroma proteome, we fractionated soluble chloroplast stroma in the non-denatured state by size exclusion chromatography with a size separation range up to ?5 MDa. To maximize protein complex stability and resolution of megadalton complexes, ionic strength and composition were optimized. Subsequent high accuracy tandem mass spectrometry analysis (LTQ-Orbitrap) identified 1081 proteins across the complete native mass range. Protein complexes and assembly states above 0.8 MDa were resolved using hierarchical clustering, and protein heat maps were generated from normalized protein spectral counts for each of the size exclusion chromatography fractions; this complemented previous analysis of stromal complexes up to 0.8 MDa (Peltier, J. B., Cai, Y., Sun, Q., Zabrouskov, V., Giacomelli, L., Rudella, A., Ytterberg, A. J., Rutschow, H., and van Wijk, K. J. (2006) The oligomeric stromal proteome of Arabidopsis thaliana chloroplasts. Mol. Cell. Proteomics 5, 114–133). This combined experimental and bioinformatics analyses resolved chloroplast ribosomes in different assembly and functional states (e.g. 30, 50, and 70 S), which enabled the identification of plastid homologues of prokaryotic ribosome assembly factors as well as proteins involved in co-translational modifications, targeting, and folding. The roles of these ribosome-associating proteins will be discussed. Known RNA splice factors (e.g. CAF1/WTF1/RNC1) as well as uncharacterized proteins with RNA-binding domains (pentatricopeptide repeat, RNA recognition motif, and chloroplast ribosome maturation), RNases, and DEAD box helicases were found in various sized complexes. Chloroplast DNA (>3 MDa) was found in association with the complete heteromeric plastid-encoded DNA polymerase complex, and a dozen other DNA-binding proteins, e.g. DNA gyrase, topoisomerase, and various DNA repair enzymes. The heteromeric ?5-MDa pyruvate dehydrogenase complex and the 0.8–1-MDa acetyl-CoA carboxylase complex associated with uncharacterized biotin carboxyl carrier domain proteins constitute the entry point to fatty acid metabolism in leaves; we suggest that their large size relates to the need for metabolic channeling. Protein annotations and identification data are available through the Plant Proteomics Database, and mass spectrometry data are available through Proteomics Identifications database. PMID:20423899

Olinares, Paul Dominic B.; Ponnala, Lalit; van Wijk, Klaas J.



Behavior of the Inadvertent pH Transient Formed by a Salt Gradient in the Ion-Exchange Chromatography of Proteins  

E-print Network

-Exchange Chromatography of Proteins Jessica Soto Pe´rez and Douglas D. Frey* Department of Chemical and Biochemical produced when a stepwise change in salt concentration is used as the eluent in ion-exchange chromatography

Frey, Douglas D.


A complete hyaluronan hydrodynamic characterization using a size exclusion chromatography-triple detector array system during in vitro enzymatic degradation.  


Size exclusion chromatography coupled with triple detection (online laser light scattering, refractometry, and viscosimetry) (SEC-TDA) was applied for the study of hyaluronan (HA) fragments produced during hydrolysis catalyzed by bovine testicular hyaluronidase (BTH). The main advantage this approach provides is the complete hydrodynamic characterization without requiring further experiments. HA was hydrolyzed using several BTH amounts and for increasing incubation times. Fragments were characterized in terms of weight and number average molecular weights (M(w) and M(n), respectively), polydispersity index (M(w)/M(n)), hydrodynamic radius (R(h)), and intrinsic viscosity ([eta]). The Mark-Houwink-Sakurada (MHS) curves (log[eta] versus logM(w)) were then derived directly. Fragments covering a whole range of M(w) (10-900kDa) and size (R(h)=4-81nm) and presenting a rather narrow distribution of molar masses (M(w)/M(n)=1.6-1.7) were produced. From the MHS curves, HA conformation resulted in a change from a random coil toward a rigid rod structure while decreasing the M(w). HA enzymatic hydrolysis in the presence of a BTH inhibitor was also monitored, revealing that inhibition profiles are affected by ionic strength. Finally, a comparison of the kinetic data derived from SEC-TDA with the data from rheological measurements suggested different strengths of the two methods in the determination of the depolymerization rate depending on the hydrolysis conditions. PMID:20399193

La Gatta, Annalisa; De Rosa, Mario; Marzaioli, Iolanda; Busico, Teresa; Schiraldi, Chiara



A hemagglutinin quantification method for development of an influenza pandemic vaccine using size exclusion high performance liquid chromatography.  


Single radial immunodiffusion (SRID) assay requires a reference antigen and an antibody to the hemagglutinin (HA) of an influenza vaccine. As it takes 2?3 months to develop the reference antigen, vaccine development is delayed in cases of an influenza pandemic. In the present study, the measurement of the HA content of influenza vaccines was assessed using size exclusion high performance liquid chromatography (SE?HPLC) for the rapid development of a pandemic vaccine. When the 2009 H1N1 reference antigen, pandemic 2009 H1N1 vaccine and 2010 seasonal influenza vaccines were analyzed by SE?HPLC, the HA of the reference antigen and vaccines was specifically separated. The presence and specificity of HA were evidenced with immunoprecipitation and ELISA assays. For the influenza vaccines, the chromatogram pattern and retention time of HA were similar among the antigen types (2009 H1N1, 2010 H3N2 and 2010 B). In addition, when SE?HPLC was applied, the ratio of HA chromatogram to peak area revealed a significant correlation with HA concentration for the reference antigen and vaccine. The result of the HA content calculation based on SE?HPLC exhibited 99.91?100% similarity, compared with that of SRID. These findings suggest that the measurement of peak area ratio/HA content using SE?HPLC may be a substitute for SRID and rapidly measure HA content to enable faster development of a vaccine during an influenza pandemic. PMID:25482872

Roh, Hang Sik; Song, Hye Min; Yun, Bo Reum; Kang, Hyun Kyung; Choi, Keum Suk; Park, Yun Ju; Kim, Dong Sub; Kim, Seung Hee; Mo, In Pil; An, Beum-Soo; Ahn, Chi Young



Simultaneous determination of protein aggregation, degradation, and absolute molecular weight by size exclusion chromatography-multiangle laser light scattering.  


The feasibility of size exclusion chromotography (SEC)-multiangle laser-light scattering as a technique to investigate aggregation and degradation of glycosylated and nonglycosylated proteins, and antibodies under various conditions such as addition of detergent, changes in pH, and variation of protein concentration and heat stress temperature was examined. Separation of proteins and their aggregates was performed using SEC-high-performance liquid chromatography. Detection of analytes was carried out with on-line UV, refractive index, and multiangle laser light-scattering detectors. Quantification and molecular weight determination were performed using commercial software. Aggregation and degradation were examined under various conditions and quantitative results are presented for bovine serum albumin, choriogonadotropin, glyceraldehyde-3-phosphate dehydrogenase, Herceptin, and ReoPro. This method can simultaneously determine both the quantities and the molecular weights of macromolecules from a single injection. The determination of molecular weight is absolute which avoids misleading results caused by molecular shape or interactions with the column matrix. This technique is valuable not only for assessing the extent of aggregation but also for effectively monitoring molecule degradation as evidenced by molecular weight reduction and change in monomer amount. PMID:16839514

Ye, Hongping



Revisiting size-exclusion chromatography for measuring structural changes in raw and pretreated mixed hardwoods and switchgrass.  


The study of the biomass porous structure and its role in defining the accessibility of cell-wall-degrading enzymes (CWDEs) to the substrate is very important for understanding the cellulase-cellulose reaction system. Specific pore volume and specific surface area are two important measures of accessibility and a variety of methods have been used to make these measurements. For this study a size exclusion chromatography system was developed to measure specific pore volume and specific surface areas for raw and pretreated mixed-hardwood and switchgrass. Polyethylene glycol (PEG) probes of known molecular diameter (1.8-13?nm) were allowed to diffuse into the pore structure of the various biomass substrate packed in the column and subsequently eluted to generate high resolution concentration measurements with excellent reproducibility. Replicate measurements of probe concentrations from this system consistently yielded coefficient of variance of less than 1.5%. Our results showed that particle size reduction had a smaller influence on the specific pore volume distribution of raw mixed-hardwoods, whereas for switchgrass the larger particles yielded a significantly lower estimate for the pore volume distribution compared to the smaller particles. Our results also clearly showed that our bi-phasic pretreatment yielded the largest increase in pore volume accessibility for mixed-hardwoods relative to switchgrass. From these results a pore size change mechanism was proposed that could explain the influence of size reduction and pretreatment on pore volume measurements. Biotechnol. Bioeng. 2015;112: 549-559. © 2014 Wiley Periodicals, Inc. PMID:25212985

Yang, Dong; Parlange, Jean-Yves; Walker, Larry P



Size exclusion chromatography and viscometry in paper degradation studies. New Mark-Houwink coefficients for cellulose in cupri-ethylenediamine.  


The paper deals with the application of size exclusion chromatography (SEC) for the studies of paper degradation phenomena. The goal is to solve some of the technical problems connected with the calibration of multi-detector SEC system and to find the correlation between SEC and viscometric results of degree of polymerization of cellulose. The results gathered for the paper samples degraded by acidic air pollutant (NO(2)) are used as an example of SEC-MALLS application. From the correlation between intrinsic viscosities and absolute value of molecular masses obtained with SEC/MALLS (Multi Angle Laser Light Scattering) technique, Mark-Houwink coefficients for cellulose in cupri-ethylenediamine solution were determined. Thus obtained coefficients were used for the determination of viscometric degree of polymerization (molecular mass) of the aged samples. An excellent correlation was found between the chromatographic values of molecular masses obtained with SEC-UV/VIS detection and the viscometric ones utilizing the improved values of Mark-Houwink coefficients. PMID:20833398

?ojewski, Tomasz; Zieba, Katarzyna; Lojewska, Joanna



Characterization of small protein aggregates and oligomers using size exclusion chromatography with online detection by native electrospray ionization mass spectrometry.  


Self-association of proteins is important in a variety of processes ranging from acquisition of native quaternary structure (where the association is tightly controlled and proceeds in a highly ordered fashion) to aggregation and amyloidosis. The latter is frequently accompanied (or indeed triggered) by the loss of the native structure, but a clear understanding of the complex relationship between conformational changes and protein self-association/aggregation remains elusive due to the great difficulty in characterizing these complex and frequently heterogeneous species. In this study, size exclusion chromatography (SEC) was used in combination with online detection by native electrospray ionization mass spectrometry (ESI MS) to characterize a commercial protein sample (serum albumin) that forms small aggregates. Although noncovalent dimers and trimers of this protein are readily detected by native ESI MS alone, combination of SEC and ESI MS allows a distinction to be made between the oligomers present in solution and those formed during the ESI process (artifacts of ESI MS). Additionally, native ESI MS detection allows a partial loss of conformation integrity to be detected across all albumin species present in solution. Finally, ESI MS detection allows these analyses to be carried out readily even in the presence of other abundant proteins coeluting with albumin. Native ESI MS as an online detection method for SEC also enables meaningful characterization of species representing different quaternary organization of a recombinant glycoprotein human arylsulfatase A even when their rapid interconversion prevents their separation on the SEC time scale. PMID:25310183

Muneeruddin, Khaja; Thomas, John J; Salinas, Paul A; Kaltashov, Igor A



High-pressure size exclusion chromatography analysis of dissolved organic matter isolated by tangential-flow ultra filtration  

USGS Publications Warehouse

A 1,000-Dalton tangential-flow ultrafiltration (TFUF) membrane was used to isolate dissolved organic matter (DOM) from several freshwater environments. The TFUF unit used in this study was able to completely retain a polystyrene sulfonate 1,800-Dalton standard. Unaltered and TFUF-fractionated DOM molecular weights were assayed by high-pressure size exclusion chromatography (HPSEC). The weight-averaged molecular weights of the retentates were larger than those of the raw water samples, whereas the filtrates were all significantly smaller and approximately the same size or smaller than the manufacturer-specified pore size of the membrane. Moreover, at 280 nm the molar absorptivity of the DOM retained by the ultrafilter is significantly larger than the material in the filtrate. This observation suggests that most of the chromophoric components are associated with the higher molecular weight fraction of the DOM pool. Multivalent metals in the aqueous matrix also affected the molecular weights of the DOM molecules. Typically, proton-exchanged DOM retentates were smaller than untreated samples. This TFUF system appears to be an effective means of isolating aquatic DOM by size, but the ultimate size of the retentates may be affected by the presence of metals and by configurational properties unique to the DOM phase.

Everett, C.R.; Chin, Y.-P.; Aiken, G.R.



Structural Evolution of Polylactide Molecular Bottlebrushes: Kinetics Study by Size Exclusion Chromatography, Small Angle Neutron Scattering and Simulations  

SciTech Connect

Structural evolution from poly(lactide) (PLA) macromonomer to resultant PLA molecular bottlebrush during ring opening metathesis polymerization (ROMP) was investigated for the first time by combining size exclusion chromatography (SEC), small-angle neutron scattering (SANS) and coarse-grained molecular dynamics (CG-MD) simulations. Multiple aliquots were collected at various reaction times during ROMP, and subsequently analyzed by SEC and SANS. The two complementary techniques enable the understanding of systematic changes in conversion, molecular weight and dispersity as well as structural details of PLA molecular bottlebrushes. CG-MD simulation not only predicts the experimental observations, but it also provides further insight into the analysis and interpretation of data obtained in SEC and SANS experiments. We find that PLA molecular bottlebrushes undergo three conformational transitions with increasing conversion (i.e., increasing the backbone length): (1) from an elongated to a globular shape due to longer side chain at lower conversion, (2) from a globular to an elongated shape at intermediate conversion caused by excluded volume of PLA side chain, and (3) the saturation of contour length at higher conversion due to chain transfer reactions.

Pickel, Deanna L [ORNL; Kilbey, II, S Michael [ORNL; Uhrig, David [ORNL; Hong, Kunlun [ORNL; Carrillo, Jan-Michael Y [ORNL; Sumpter, Bobby G [ORNL; Ahn, Suk-Kyun [ORNL; Han, Youngkyu [ORNL; Kim, Dr. Tae-Hwan [Korea Atomic Energy Research Institute; Smith, Gregory Scott [ORNL; Do, Changwoo [ORNL



Analysis of a complex polysaccharide (gum arabic) by multi-angle laser light scattering coupled on-line to size exclusion chromatography and flow field flow fractionation  

Microsoft Academic Search

The heterogeneous polysaccharide gum arabic has been characterized using size exclusion chromatography (SEC) and flow field flow fractionation (F4) coupled on-line to multi-angle laser light scattering (MALLS). Two distinct populations have been shown. About 80% of the material consist of highly branched arabinogalactan (AG) units. The rest is mainly composed of heterogeneous arabinogalactan–protein complex (AGP) of high molecular weight. The

L. Picton; I. Bataille; G. Muller



The formation of heat-induced protein aggregates in whey protein\\/pectin mixtures studied by size exclusion chromatography coupled with multi-angle laser light scattering detection  

Microsoft Academic Search

The molecular weight (MW) distribution of whey proteins (WP) mixed with various pectins was investigated using size exclusion chromatography (HPSEC) combined with multi-angle laser light scattering (MALLS). Samples were prepared with 1, 2, 5, 8% WP and 1% pectin. Mixtures were studied at pH 6.0 and CaCl2 concentrations of 0, 5, 10mM. Aggregate formation was determined after heating at various

Martin Beaulieu; Milena Corredig; Sylvie L. Turgeon; Louise Wicker; Jean-Louis Doublier



Absolute Molecular Weight Distribution of Low-Molecular-Weight Heparins by Size-Exclusion Chromatography with Multiangle Laser Light Scattering Detection  

Microsoft Academic Search

The absolute molecular weight (Mr) distribution of seven low-molecular-weight (LMW) heparin products was determined by size-exclusion chromatography (SEC) coupled with multiangle laser light scattering (MALLS) detection. The SEC\\/MALLS technique does not rely on relativeMrstandards for column calibration and yields absoluteMrestimates directly from the angular dependence of scattered light intensity as a function of concentration, as formulated by light scattering theory.

James E. Knobloch; Patrick N. Shaklee



Use of Multi-Angle Laser Light Scattering and Size-Exclusion Chromatography to Characterize the Molecular Weight and Types of Aggregates Present in Commercial Whey Protein Products  

Microsoft Academic Search

In this study, a range of commercial whey protein products were characterized by the use of size-exclusion chromatography coupled with a multi-angle laser light scattering (MALLS) detector. The MALLS system de- tected some very large-sized material that eluted close to void volume in all samples; this material was hardly detected by concentration detector. It was demon- strated by chitosan treatment

T. Wang; J. A. Lucey



Characterization of a polysaccharide–protein complex from Ganoderma tsugae mycelium by size-exclusion chromatography combined with laser light scattering  

Microsoft Academic Search

A water-soluble polysaccharide–protein complex (GM3) extracted from the mycelium of Ganoderma tsugae was characterized using size-exclusion chromatography combined with laser light scattering (SEC-LLS). Two peaks coded as fractions I and II appeared in the SEC pattern of GM3 in 0.5 M NaCl aqueous solution, corresponding to the weight-average molecular mass (Mw) of 355×104 and 6.3×104, respectively. The relationship between the

Yanfei Peng; Lina Zhang



Size-exclusion chromatography–multiangle laser light scattering analysis of ?-lactoglobulin and bovine serum albumin in aqueous solution with added salt  

Microsoft Academic Search

The solution characteristics of ?-LGB (?-lactoglobulin) and BSA (bovine serum albumin) are reported as determined by size-exclusion chromatography with on-line multiangle laser light scattering, differential refractive index and UV detection. The order of the three in series placed detectors as well as the interdetector volumes have been carefully pointed out. At concentrations below 2.5 mg\\/ml and at different values of

J. A. P. P van Dijk; J. A. M Smit



A novel size-exclusion high performance liquid chromatography (SE-HPLC) method for measuring degree of amylose retrogradation in rice starch  

Microsoft Academic Search

The objective of this study was to develop a new method to measure the degree of amylose retrogradation in rice starch using size-exclusion high performance liquid chromatography (SE-HPLC). This developed method is based on the change in molecule size related to the retention time of amylose. Results showed that SE-HPLC was able to provide accurate data to evaluate the amylose

Yaoqi Tian; Yin Li; Xueming Xu; Zhengyu Jin; Aiquan Jiao; Jinpeng Wang; Bo Yu



Size-Exclusion Chromatography with On-Line Light-Scattering, Absorbance, and Refractive Index Detectors for Studying Proteins and Their Interactions  

Microsoft Academic Search

Techniques of using size-exclusion chromatography (SEC) with on-line light-scattering, uv absorbance, and refractive index detectors to characterize the polypeptide molecular weights of simple proteins or glycoproteins or to determine the stoichiometry of protein complexes are described. Two unique advantages of this approach over conventional SEC are that the molecular weight measurement is independent of elution position and can exclude the

Jie Wen; Tsutomu Arakawa; John S. Philo



Sample preparation and reversed phase-high performance liquid chromatography analysis of food-derived peptides  

Microsoft Academic Search

Reversed phase-high performance liquid chromatography (RP-HPLC) is an indispensable technique in food peptide research. This paper briefly reviews the methods for isolation and RP-HPLC analysis of food-derived peptides. A critical aspect is the sample preparation and clean-up. For this, selective precipitation, ultrafiltration, size exclusion chromatography, ion-exchange chromatography and solid phase extraction (SPE) are used. SPE with non-polar or ion-exchange cartridges,

T. Herraiz



Method for the determination of dissolved chloride, nitrate, and sulfate in natural water using ion chromatography  

USGS Publications Warehouse

Ion chromatography was used for the determination of dissolved chloride, nitrate and sulfate in natural water where concentrations ranged from a detection limit of 0.02 milligrams per liter to 80 milligrams per liter for chloride, to 18 milligrams per liter for nitrate, and to 280 milligrams per liter for sulfate. Specific conductance was the mode of detection used. Three analytical sample size loops of 11, 61, and 250 microliters, were used to include the analytical ranges described. U.S. Geological Survey Standard Reference Water Samples were analyzed to test the precision and accuracy of the analyses.

Brinton, Terry I.; Antweiler, Ronald C.; Taylor, Howard E.



Tailored Noise Waveform/ Collision-Induced Dissociation of Ions Stored in a Linear Ion Trap Combined with Liquid Chromatography/Fourier Transform Ion Cyclotron Resonance Mass Spectrometry  

SciTech Connect

A new collision-induced dissociation (CID) technique based on broadband tailored noise waveform (TNW) excitation of ions stored in a linear ion trap has been developed. In comparison with the conventional sustained off-resonance irradiation (SORI) CID method commonly used in Fourier transform ion cyclotron resonance mass spectrometry, this MS/MS technique increases throughput by eliminating the long pump-down delay associated with gas introduction into the high vacuum ICR cell region. In addition, the TNW-CID method speeds spectrum acquisition since it does not require Fourier transformation, calculation of resonant frequencies and generation of the excitation waveforms. We demonstrate TNW-CID coupled with on-line capillary reverse phase liquid chromatography separations for identification of peptides. The experimental results are compared with data obtained using conventional quadrupole ion trap MS/MS and SORI-CID MS/MS in an ICR cell.

Vilkov, Andrey N.; Bogdanov, Bogdan; Pasa-Tolic, Liljiana; Prior, David C.; Anderson, Gordon A.; Masselon, Christophe D.; Moore, Ronald J.; Smith, Richard D.



Liquid chromatography ion trap/time-of-flight mass spectrometric study on the fragmentation of an acetildenafil analogue.  


Liquid chromatography ion-trap time-of-flight mass spectrometry was employed to elucidate the fragmentation pathways of an analogue of acetildenafil. Based on the accurate masses of the parent ion, product ions and neutral losses of acetildenafil analogue, its fragmentation pathways were proposed. The information is useful for the on-line structural identification of unknown analogues of acetildenafil found as adulterants in herbal products. PMID:17881792

Zou, Peng; Oh, Sharon Sze-Yin; Kiang, Kin-Har; Low, Min-Yong; Koh, Hwee-Ling



Portable, fully autonomous, ion chromatography system for on-site analyses.  


The basic operating principles of a portable, fully autonomous, ion chromatography system are described. The system affords the user the ability to collect and analyze samples continuously for 27 days, or about 1930 injections before needing any user intervention. Within the 13 kg system, is a fully computer controlled autosampling, chromatography and data acquisition system. An eluent reflux device (ERD), which integrates eluent suppression and generation in a single multi-chambered device, is used to minimize eluent consumption. During operation, about 1 ?L of water per minute is lost to waste while operating standard-bore chromatography at 0.5 mL min(-1) due to eluent refluxing. Over the course of 27 days, about 100mL of rinse water is consumed, effectively eliminating waste production. Data showing the reproducibility (below 1% relative standard deviation over 14 days) of the device is also presented. Chromatographic analyses of common anions (Cl(-), NO3(-), SO4(2-), PO4(3-)), is accomplished in under 15 min using a low backpressure guard column with ? 25 mM KOH isocratic elution. For detection, a small capacitively-coupled contactless conductivity detector (C4D) is employed, able to report analytes in the sub to low micromolar range. Preconcentration of the injected samples gives a 50-fold decrease in detection limits, primarily utilized for in-situ detection of phosphate (LOQ 10 ?g L(-1)). Field analyses are shown for multiple on-site analyses of stream water indifferent weather conditions. PMID:24913366

Elkin, Kyle R



Comparison of Diafiltration and Size-Exclusion Chromatography to Recover Hemicelluloses From Process Water From Thermomechanical Pulping of Spruce  

NASA Astrophysics Data System (ADS)

Hemicelluloses constitute one of the most abundant renewable resources on earth. To increase their utilization, the isolation of hemicelluloses from industrial biomass side-streams would be beneficial. A method was investigated to isolate hemicelluloses from process water from a thermomechanical pulp mill. The method consists of three steps: removal of solids by microfiltration, preconcentration of the hemicelluloses by ultrafiltration, and purification by either size-exclusion chromatography (SEC) or diafiltration. The purpose of the final purification step is to separate hemicelluloses from small oligosaccharides, monosaccharides, and salts. The ratio between galactose, glucose, and mannose in oligo- and polysaccharides after preconcentration was 0.8?1?2.8, which is similar to that found in galactoglucomannan. Continuous diafiltration was performed using a composite fluoro polymer membrane with cutoff of 1000 Da. After diafiltration with four diavolumes the purity of the hemicelluloses was 77% (gram oligo- and polysaccharides/ gram total dissolved solids) and the recovery was 87%. Purification by SEC was performed with 5, 20, and 40% sample loadings, respectively and a flow rate of 12 or 25 mL/min (9 or 19 cm/h). The purity of hemicelluloses after SEC was approx 82%, and the recovery was above 99%. The optimal sample load and flow rate were 20% and 25 mL/min, respectively. The process water from thermomechanical pulping of spruce is inexpensive. Thus, the recovery of hemicelluloses is not of main importance. If the purity of 77%, obtained with diafiltration, is sufficient for the utilization of the hemicelluloses, diafiltration probably offers a less expensive alternative in this application.

Andersson, Alexandra; Persson, Tobias; Zacchi, Guido; Stålbrand, Henrik; Jönsson, Ann-Sofi


The removal of uranium from acidic media using ion exchange and/or extraction chromatography  

SciTech Connect

The separation and purification of uranium from either nitric acid or hydrochloric acid media can be accomplished by using either solvent extraction or ion-exchange. Over the past two years at Los Alamos, emerging programs are focused on recapturing the expertise required to do limited, small-quantity processing of enriched uranium. During this period of time, we have been investigating ion-addition, waste stream polishing is associated with this effort in order to achieve more complete removal of uranium prior to recycle of the acid. Extraction chromatography has been demonstrated to further polish the uranium from both nitric and hydrochloric acid media thus allowing for a more complete recovery of the actinide material and creation of less waste during the processing steps.

FitzPatrick, J.R.; Schake, B.S.; Murphy, J.; Holmes, K; West, M.H.



Use of ion chromatography for the determination of heavy and transition metals in biochemical samples.  


A novel, highly sensitive method for simultaneous separation and determination of Cu2+, Ni2+, Zn2+, Cd2+, Co2+, Mn2+ and Pb2+ in biochemical samples was developed and evaluated by ion chromatography. All of these metals were well separated on a bifunctional ion-exchange column by a concentration gradient of oxalic acid and sodium chloride eluents, coupled with spectrophotometric detection after post-column derivatization with 2-[(5-bromo-2-pyridyl)azo]-5-diethylaminophenol at 560 nm. The method detection limits (signal-to-noise 3:1) were at microg l(-1) levels. The calibration graphs were linear (r2>0.999) over two-orders of magnitude. This technique was optimized and validated by analyzing five standard biochemical references. PMID:10536855

Lu, H; Mou, S; Riviello, J M



[Nano-electrospray ionization-ion mobility spectrometry and its combination with high performance liquid chromatography].  


A nano-electrospray ionization-ion mobility spectrometer (nanoESI-IMS) was built up. Firstly, the effects of the parameters such as drift gas flow rate and solvent flow rate on the desolvation capability were studied and optimized. Then, a series of compounds were used to characterize the nanoESI-IMS system. The results showed that, complete desolvation was achieved for nano-electrospray ion droplets with the nanoESI-IMS apparatus. The limit of detection of this instrument for trioctylamine could reach 10 microg/L. Finally, this instrument was coupled to the high performance liquid chromatography (HPLC) as a detector for amines analysis. A test mixture containing triethylamine, diethylamine and butylamine was successfully separated and determined by the HPLC-nanoESI-IMS system. Linear response ranges of about two orders of magnitude were achieved for triethylamine, diethylamine and butylamine with this system. PMID:23898640

Chen, Chuang; Wang, Weiguo; Liang, Xixi; Zhou, Qinghu; Peng, Liying; Wen, Meng; Ju, Bangyu; Zhao, Kun; Liu, Jun; Li, Haiyang



Determination of anionic surfactants during wastewater recycling process by ion pair chromatography with suppressed conductivity detection  

NASA Technical Reports Server (NTRS)

A direct approach utilizing ion pairing reversed-phase chromatography coupled with suppressed conductivity detection was developed to monitor biodegradation of anionic surfactants during wastewater recycling through hydroponic plant growth systems and fixed-film bioreactors. Samples of hydroponic nutrient solution and bioreactor effluent with high concentrations (up to 120 mS electrical conductance) of inorganic ions can be analyzed without pretreatment or interference. The presence of non-ionic surfactants did not significantly affect the analysis. Dynamic linear ranges for tested surfactants [Igepon TC-42, ammonium lauryl sulfate, sodium laureth sulfate and sodium alkyl (C10-C16) ether sulfate] were 2 to approximately 500, 1 to approximately 500, 2.5 to approximately 550 and 3.0 to approximately 630 microg/ml, respectively.

Levine, L. H.; Judkins, J. E.; Garland, J. L.; Sager, J. C. (Principal Investigator)



Quantification and characterization of dissolved organic nitrogen in wastewater effluents by electrodialysis treatment followed by size-exclusion chromatography with nitrogen detection.  


Dissolved organic nitrogen (DON) can act as a precursor of nitrogenous disinfection byproducts during oxidative water treatment. Quantification and characterization of DON are still challenging for waters with high concentrations of dissolved inorganic nitrogen (DIN, including ammonia, nitrate and nitrite) relative to total dissolved nitrogen (TDN) due to the cumulative analytical errors of independently measured nitrogen species (i.e., DON = TDN - NO2(-) - NO3(-) - NH4(+)/NH3) and interference of DIN species to TDN quantification. In this study, a novel electrodialysis (ED)-based treatment for selective DIN removal was developed and optimized with respect to type of ion-exchange membrane, sample pH, and ED duration. The optimized ED method was then coupled with size-exclusion chromatography with organic carbon, UV, and nitrogen detection (SEC-OCD-ND) for advanced DON analysis in wastewater effluents. Among the tested ion-exchange membranes, the PC-AR anion- and CMT cation-exchange membranes showed the lowest DOC loss (1-7%) during ED treatment of a wastewater effluent at ambient pH (8.0). A good correlation was found between the decrease of the DIN/TDN ratio and conductivity. Therefore, conductivity has been adopted as a convenient way to determine the optimal duration of the ED treatment. In the pH range of 7.0-8.3, ED treatment of various wastewater effluents with the PC-AR/CMT membranes showed that the relative residual conductivity could be reduced to less than 0.50 (DIN removal >90%; DIN/TDN ratio ? 0.60) with lower DOC losses (6%) than the previous dialysis and nanofiltration methods (DOC loss >10%). In addition, the ED method is shorter (0.5 h) than the previous methods (>1-24 h). The relative residual conductivity was further reduced to ? 0.20 (DIN removal >95%; DIN/TDN ratio ? 0.35) by increasing the ED duration to 0.7 h (DOC loss = 8%) for analysis by SEC-OCD-ND, which provided new information on distribution and ratio of organic carbon and nitrogen in different molecular weight fractions of effluent organic matter. PMID:23916154

Chon, Kangmin; Lee, Yunho; Traber, Jacqueline; von Gunten, Urs



Effect of injection matrix concentration on peak shape and separation efficiency in ion chromatography.  


In HPLC, injection of solvents that differ from the eluent can result in peak broadening due to solvent strength mismatch or viscous fingering. Broadened, distorted or even split analyte peaks may result. Past studies of this injection-induced peak distortion in reversed phase (RPLC) and hydrophilic interaction (HILIC) liquid chromatography have led to the conclusion that the sample should be injected in the eluent or a weaker solvent. However, there have been no studies of injection-induced peak distortion in ion chromatography (IC). To address this, injection-induced effects were studied for six inorganic anions (F(-), Cl(-), NO2(-), Br(-), NO3(-) and SO4(2-)) on a Dionex AS23 IC column using a HCO3(-)/CO3(2-) eluent. The VanMiddlesworth-Dorsey injection sensitivity parameter (s) showed that IC of anions has much greater tolerance to the injection matrix (HCO3(-)/CO3(2-) herein) mismatch than RPLC or HILIC. Even when the injection contained a ten-fold greater concentration of HCO3(-)/CO3(2-) than the eluent, the peak shapes and separation efficiencies of six analyte ions did not change significantly. At more than ten-fold greater matrix concentrations, analyte anions that elute near the system peak of the matrix were distorted, and in the extreme cases exhibited a small secondary peak on the analyte peak front. These studies better guide the degree of dilution needed prior to IC analysis of anions. PMID:25456596

Zhang, Ya; Lucy, Charles A



Computer-assisted multi-segment gradient optimization in ion chromatography.  


This study reports simulation and optimization of ion chromatography separations using multi-segment gradient elution. First, an analytical expression for the gradient retention factor under these complex elution profiles was derived. This allows a rapid retention time prediction calculations under different gradient conditions, during computer-assisted method development. Next, these analytical expressions were implemented in an in-house written Matlab(®) routine that searches for the optimal (multi-segment) gradient conditions, either via a four-segment grid search or via the recently proposed one-segment-per-component search, in which the slope is adjusted after the elution of each individual component. Evaluation of the retention time simulation and optimization approaches was performed on a mixture of 18 inorganic anions and different subsets with varying number of compounds. The two considered multi-segment gradient optimization searches resulted in similar proposed gradient profiles, and corresponding chromatograms. Moreover, the resultant chromatograms were clearly superior to the chromatograms obtained from the best simple linear gradient profiles, found via a fine grid search. The proposed approach is useful for automated method development in ion chromatography in which complex elution profiles are often used to increase the separation power. PMID:25596760

Tyteca, Eva; Park, Soo Hyun; Shellie, Robert A; Haddad, Paul R; Desmet, Gert



Quantitation of diethylene glycol and its metabolites by gas chromatography mass spectrometry or ion chromatography mass spectrometry in rat and human biological samples.  


The misuse of the commonly used chemical diethylene glycol (DEG) has lead to many poisonings worldwide. Methods were developed for analysis of DEG and its potential metabolites; ethylene glycol, glycolic acid, oxalic acid, diglycolic acid and hydroxyethoxy acetic acid in human urine, serum and cerebrospinal fluid samples, collected following a DEG-associated poisoning in the Republic of Panama during 2006. In addition, methods were developed for rat blood, urine, kidney and liver tissue to support toxicokinetic analysis during the conduct of DEG acute toxicity studies in the rat. Sample analysis was conducted using two techniques; ion chromatography with suppressed conductivity and negative ion electrospray ionization with MS detection or with gas chromatography using electron impact ionization or methane negative chemical ionization with MS detection. Stable-isotope-labeled analogs of each analyte were employed as quantitative internal standards in the assays. PMID:24668490

Perala, Adam W; Filary, Mark J; Bartels, Michael J; McMartin, Kenneth E



The use of ion interaction chromatography with a post-column colorimetric reaction to determine bromate in water  

SciTech Connect

Ozonation of slightly alkaline water can oxidize endogenous bromide to bromate - a considerable concern for drinking water. We have developed a method for bromate based upon an adaption of ion-pairing, reversed-phase chromatography, which is also known as ion interaction chromatography - a name that some authorities believe more accurately describes the mechanism of the chromatographic separation (H. Small, Ion Chromatography, pp 106-118, Plenum Press, 1989). A colorimetric reaction based upon the oxidation of o-dianisidine provides post-column determination of bromate. Iodate, chlorite, and nitrite are chromatographically resolved from bromate and determined by the same o-dianisidine post-column reaction. With filtration as the only required pretreatment, the method can be used to determine bromate at 5 ug/L in drinking water.

Warner, C.R.; Daniels, D.H.; Joe, F.L. Jr.; Diachenko, G.W. [Food and Drug Administration, Washington, DC (United States)



Direct Determination of Trace-Level Haloacetic Acids in Drinking Water by Two-1 Dimensional Ion Chromatography with Suppressed Conductivity2  

E-print Network

Chromatography with Suppressed Conductivity2 3 DominiqueVerreya,b, *, Mari-VorganLouyera,b, Olivier Thomasa,GC/MS with derivatization and LC/MS, UV or conductivity. A new method, based22 on two-dimensional ion chromatography (IC 2D to quantify HAA at trace levels in drinking water.31 32 Keywords: ionic chromatography; two dimensions

Boyer, Edmond


Separation of tricyclic psychosedative drugs by high-speed ion-pair partition and liquid-solid adsorption chromatography.  


The application of high-speed ion-pair partition and liquid-solid adsorption chromatography to the separation of twenty common tricyclic tranquilizers and antidepressant drugs is described. In the ion-pair system, amine-perchlorate ion-pairs were extracted from an aqueous stationary phase supported on 10-mum silica gel by organic eluents containing a chloromethane and a higher aliphatic alcohol, and chromatographic parameters for elution by eight eluent mixtures are presented. Using 5 mm times 120 mm columns good separations, according to chemical class, were achieved. For adsorption chromatography, the components were eluted from 20-mum spherical alumina using eluents containing methylene chloride, n-hexane or n-pentane, and acetic acid. Chromatographic parameters are given for eight eluent compositions. Components differing little in structure are well separated by liquid-solid adsorption chromatography. Compared with ion-pair partition chromatography, adsorption chromatography is much more selective for compounds of the same chemical type. The two methods are therefore complementary. Both methods gave plate heights in the range of 0.1 to 0.3 mm. PMID:234973

Knox, J H; Jurand, J



Using contemporary liquid chromatography theory and technology to improve capillary gradient ion-exchange separations.  


The gradient-performance limits of capillary ion chromatography have been assessed at maximum system pressure (34.5 MPa) using capillary columns packed with 4.1 ?m macroporous anion-exchange particles coated with 65 nm positively-charged nanobeads. In analogy to the van-Deemter curve, the gradient performance was assessed applying different flow rates, while decreasing the gradient time inversely proportional to the increase in flow rate in order to maintain the same retention properties. The gradient kinetic-performance limits were determined at maximum system pressure, applying tG/t0=5, 10, and 20. In addition, the effect of retention on peak width was assessed in gradient mode for mono-, di-, and trivalent inorganic anions. The peak width of late-eluting ions can be significantly reduced by using concave gradient, resulting in better detection sensitivity. A signal enhancement factor of 8 was measured for a late-eluting ion when applying a concave instead of a linear gradient. For the analysis of a complex anion mixture, a coupled column with a total length of 1.05 m was operated at the kinetic-performance limit applying a linear 250 min gradient (tG/t0=10). The peak capacity varied between 200 and 380 depending on analyte retention, and hence on charge and size of the ion. PMID:25454130

Wouters, Bert; Broeckhoven, Ken; Wouters, Sam; Bruggink, Cees; Agroskin, Yury; Pohl, Christopher A; Eeltink, Sebastiaan



Continuous measurement of macronutrient ions in the transpiration stream of intact plants using the meadow spittlebug coupled with ion chromatography.  


A method is described for continuous, nondestructive analysis of xylem-borne mineral nutrients in intact transpiring plants. The method uses the xylem-feeding insect the meadow spittlebug (Philaenus spumarius L. [Homoptera: Cercopidae]). This insect will feed from a wide range of plant species and organs. Insect excreta can be collected at all times of the day and night, and its mineral ion content can be analyzed rapidly, and without purification, by ion chromatography. The excreta will have a mineral content virtually identical to that of xylem sap. Cages suitable for containing the insects and collecting excreta from any desired location on plants in both laboratory and greenhouse are described. Even in the greenhouse, evaporation had only a minor effect on the sample ion content. Example results are presented which illustrate dynamics, over several days, in the xylem concentrations of sodium (Na(+)), potassium (K(+)), NH(4)(+), magnesium (Mg(2+)), calcium (Ca(2+)), chloride (Cl(-)), NO(3)(-), PO(4)(3-), and SO(4)(2-). These data were collected from young plants growing in pots of compost in the laboratory and from fully mature pepper (Capsicum annuum L. cv Bellboy) plants growing in hydroponics (rockwool) in the greenhouse. This method should facilitate studies of macronutrient uptake and transport in a range of plants and environments. PMID:12428008

Malone, Michael; Herron, Michelle; Morales, M-Angeles



Method for Characterization of Low Molecular Weight Organic Acids in Atmospheric Aerosols Using Ion Chromatography Mass  

E-print Network

Chromatography Mass Spectrometry Lacey C. Brent,* Jessica L. Reiner, Russell R. Dickerson, and Lane C. Sander spectroscopy (FTIR), liquid chromatography with ultraviolet detection (LCUV), or nuclear magnetic resonance chromatography mass spectrometry (GCMS) methods have demonstrated reliable structural identification of water

Dickerson, Russell R.


Determination of bromate in sea water using multi-dimensional matrix-elimination ion chromatography.  


A multi-dimensional matrix-elimination ion chromatography approach has been applied to the determination of bromate in seawater samples. Two-dimensional and three-dimensional configurations were evaluated for their efficacy to eliminate the interference caused by the high concentration of ubiquitous ions present in seawater, such as chloride and sulfate. A two-dimensional approach utilising a high capacity second dimension separation comprising two Dionex AS24 columns connected in series was applied successfully and permitted the determination of bromate in undiluted seawater samples injected directly onto the ion chromatography system. Using this approach the limit of detection (LOD) for bromate based on a signal to noise ratio of 3 was 1050 ?g/L using a 500 ?L injection loop. Good linearity was obtained for bromate with correlation coefficients for the calibration curves of 0.9981 and 0.9996 based on peak height and area, respectively. A three-dimensional method utilising two 10-port switching valves to allow sharing of the second suppressor and detector between the second and third dimension separations showed better resolution and detection for bromate and reduced the LOD to 60 ?g/L for spiked seawater samples. Good linearity was maintained with correlation coefficients of 0.9991 for both peak height and area. Ozonated seawater samples were also analysed and exhibited a non-linear increase in bromate level on increasing ozonation time. A bromate concentration in excess of 1770 ?g/L was observed following ozonation of the seawater sample for 120 min. Recoveries for the three-dimensional system were 92% and 89% based on peak height and area, respectively, taken over 5 ozonated samples with 3 replicates per sample. PMID:22074647

Zakaria, Philip; Bloomfield, Carrie; Shellie, Robert A; Haddad, Paul R; Dicinoski, Greg W



Size Exclusion Chromatography Studies of the Initial Self-Association Steps of Chicken Egg White Lysozyme Nucleation  

NASA Technical Reports Server (NTRS)

Nucleation is one of the least understood aspects of crystallogenesis. In the case of macromolecule nucleation, this understanding is further hampered by uncertainty over what precisely is being discussed. We define the process of solute self-association (aggregation, oligomerization, interaction, clustering, etc.) whereby n-mers (n > or = 2) having a crystallographic or nascent crystallographic arrangement leading to the critical nucleus reversibly form in the solution, to be part of the nucleation process. This reversible self-association process is a fundamental part of the nucleation process, and occurs as a function of the solute concentration. In the case of chicken egg white lysozyme, a considerable body of experimental evidence leads us to the conclusion that it also forms the crystal growth units. Size exclusion chromatography is a simple and direct method for determining the equilibrium constants for the self-association process. A Pharmacia FPLC system was used to provide accurate solution flow rates. The column, injection valve, and sample loop were all mounted within a temperature-controlled chamber. Chromatographically re-purified lysozyme was first dialyzed against the column equilibration buffer, with injection onto the column after several hours pre-incubation at the running temperature. Preliminary experiments, were carried out using a Toyopearl HW-50F column (1 x 50cm), equilibrated with 0.1 M sodium acetate, 5% sodium chloride, pH 4.6, at 15C. Protein concentrations from 0.1 to 4 mg/ml were employed (C(sub sat) = 1.2 mg/ml). The data from several different protein preparations consistently shows a progressively decreasing elution volume with increasing protein concentration, indicating that reversible self-association is occurring. The dotted line indicates the monomeric lysozyme elution volume. However, lysozyme interacts with the column matrix in these experiments, which complicates data analysis.Accordingly, we are testing silica-based HPLC columns in an effort to eliminate this problem and substantially reduce the column volume and experimental run time. The results and data analysis from these and subsequent experiments will be presented.

Ewing, Felecia; Donovan, David; Pusey, Marc



Insight into the distribution of molecular weights and higher-order structure of hyaluronans and some beta-(1 --> 3)-glucans by size exclusion chromatography.  


The effects of high energy ultrasound and slightly raised temperature combined with the denaturing action of dimethylsulphoxide on the molecular weight and higher-order structure of hyaluronans and some beta-(1 --> 3)-glucans were studied by means of size exclusion chromatography (SEC) technique. Some experimental conditions connected with the (bio-)polymer sample preparation prior to its SEC analysis are overviewed in the light of informational relevance of studies where the action of a physical and/or chemical agent changes the hydrodynamic size of the m omolecule. PMID:8924726

Soltés, L; Mislovicová, D; Sébille, B



[Determination of alditols in foods by ion chromatography-mass spectrometry].  


A method for the determination of alditols in foods by ion chromatography-mass spectrometry (IC-MS) has been developed. The samples were extracted and cleaned up with the solid phase extraction (SPE). Then, the ion chromatographic separation was performed on a CarboPar MA1 column. The alditols were determined by MS with the selected ion monitoring (SIM) mode and quantified by the external standard method. The calibration curves showed good linearity in the certain ranges with the correlation coefficients (R2) greater than 0.99. The limits of quantification (S/N = 10) of erythritol, xylitol, D-sorbitol, D-mannitol, lactitol, maltitol were 0.98, 1.99, 2.24, 5.92, 13.56, 13.21 mg/kg and the limits of detection (S/N = 3) were 0.28, 0.59, 0.71, 1.74, 4.14, 4.03 mg/kg, respectively. The spiked recoveries of the alditols in the foods at different levels were in the range of 82.5%-108.0% with the relative standard deviations (RSDs) of 1.5%-7.6%. The sensitivity, accuracy and precision of the method meet the technical standards of the determination. The method can be applied to the determination of alditols in foods. PMID:24558846

Zhou, Hongbin; Xiong, Zhiyu; Li, Ping; Li, Jing; Sun, Li; Zhao, Yunxia



Determination of trace metallic impurities in high-purity quartz by ion chromatography.  


A method has been developed for the determination of relevant trace impurities (alkali, alkaline and transition metals) in high purity quartz by ion-chromatography. In situ reagent (HF) purification and simultaneous sample dissolution was achieved in a multichannel vapour phase digestion assembly. Twenty-one samples can be digested at a time in this vapour phase system. Significant decrease in the process blank levels for all the analytes was observed. Drastic reduction (250 times) of NH4+ blank was achieved in the described vapour phase digestion, which enables the determination of trace concentration of sodium in high purity quartz. After volatilisation of the matrix and unreacted HF, the clear water leached solutions were injected into an ion-chromatograph equipped with conductivity detector for the determination of alkali and alkaline earth metals. In the case of transition metals, the trace residues were leached with 10 mM HCl and after separation on a mixed bed analytical column (IonPac CS5) were detected by spectrophotometry after post column derivatisation using 4-(2-pyridylazo)resorcinol (PAR). The accuracy of the result was checked by their comparison with those obtained by independent methods like inductively coupled plasma (ICP) MS and ICP atomic emission spectrometry. The achievable detection limits are between 0.4 ng/g (Li) and 22 ng/g (Mn). The application of the method to the determination of the above trace metals in two high-purity-grade quartz samples is demonstrated. PMID:14753768

Dash, K; Chandrasekaran, K; Thangavel, S; Dhaville, S M; Arunachalam, J



Simultaneous and sensitive analysis of aliphatic carboxylic acids by ion-chromatography using on-line complexation with copper(II) ion.  


A new approach to ion chromatography is proposed to improve the UV detection of aliphatic carboxylic acids separated by anion-exchange chromatography. When copper(II) ion added to the mobile phase, it forms complexes with carboxylic acids that can be detected at 240nm. The absorbance was found to increase with increasing copper(II) ion concentration. The retention times of ?-hydroxy acids were also found to depend on the copper(II) ion concentration. Addition of acetonitrile to the mobile phase improved the separation of aliphatic carboxylic acids. The detection limits of the examined carboxylic acids (formate, glycolate, acetate, lactate, propionate, 3-hydroxypropionate, n-butyrate, isobutyrate, n-valerate, isovalerate, n-caproate) calculated at S/N=3 ranged from 0.06 to 3?M. The detector signal was linear over three orders of magnitude of carboxylic acid concentration. The proposed method was successfully applied to analyze aliphatic carboxylic acids in rainwater and bread. PMID:25523885

Kemmei, Tomoko; Kodama, Shuji; Yamamoto, Atsushi; Inoue, Yoshinori; Hayakawa, Kazuichi



Rapid measurement of free cyanide in liquor by ion chromatography with pulsed amperometric detection.  


This study investigated the measurement of free cyanide in liquor by ion chromatography coupled with pulsed amperometric detection (IC-PAD). Eluent concentration, interferent evaluation and method performance were discussed. Results show that free cyanide in liquor can be rapidly determined by the optimised IC-PAD method. A sample requires only 1:100 dilution and simple filtration before being subjected to IC-PAD. The linear range is 1-5000?g/L with an R value of 0.9998. The detection limit is 1?g/L for a 25?L injection loop. The overall relative standard deviation (RSD) of the method is less than 5%, and the recovery range is from 98.1% to 105.0%. This study has been proven significant and may have potential applications in liquors analysis. PMID:25442607

Wu, Wenlin; Xiao, Quanwei; Zhang, Ping; Ye, Mei; Wan, Yuping; Liang, Hengxing



Monitoring of Lactobacillus fermentation process by using ion chromatography with a series piezoelectric quartz crystal detector.  


A new method monitoring Lactobacillus fermentation process, which combines ion chromatography (IC) with a series piezoelectric quartz crystal (SPQC) technique, is presented in this paper. Monitoring of the fermentation process was realized by examining the rate of production of lactic acid. An automatic membrane dialyser was used for the pretreatment of the sample in on-line monitoring. A mixture of p-hydroxybenzoic acid and N,N-diethylethanolamine was adopted as mobile phase and its flow rate was 0.8 ml/min. The effects of some fermentation conditions were also discussed in detail. Accordingly, the optimal fermentation conditions were obtained. This method is simple and convenient while the results obtained are accurate and reliable. PMID:11165339

Zhang, J; Xie, Y; Dai, X; Wei, W



Isotope effects of neodymium in different ligands exchange systems studied by ion exchange displacement chromatography  

PubMed Central

The isotope effects of neodymium in Nd-glycolate ligand exchange system were studied by using ion exchange chromatography. The separation coefficients of neodymium isotopes, ?’s, were calculated from the observed isotopic ratios at the front and rear boundaries of the neodymium adsorption band. The values of separation coefficients of neodymium isotopes, ?’s, for the Nd-glycolate ligand exchange system were compared with those of Nd-malate and Nd-citrate, which indicated that the isotope effects of neodymium as studied by the three ligands takes the following direction Malate > Citrate > Glycolate. This order agrees with the number of available sites for complexation of each ligand. The values of the plate height, HETP of Nd in Nd-ligand exchange systems were also calculated.

Ismail, Ibrahim; Fawzy, Ahmed S.; Ahmad, Mohammad I.; Aly, Hisham F.; Nomura, Masao; Fujii, Yasuhiko



Determination of biogenic amines in fish tissues by ion-exchange chromatography with conductivity detection.  


Some biogenic amines, such as putrescine, cadaverine, spermidine and histamine, have been found to be useful as quality indices for the decomposition of fish, so research on the simultaneous analysis of various biogenic amines in food is of interest and importance. The intake of histamine may cause an allergic intoxication known as "scombroid poisoning" while secondary biogenic amines can potentiate the toxicity of histamine and in addition can react with nitrite to form carcinogenic nitrosamines. A new method for the simultaneous determination of underivatized biogenic amines based on ion-exchange chromatography with conductivity detector has been developed. The proposed method offers a number of advantages over previous pulsed amperometric detection and integrated pulsed amperometric detection methods such as simpler extraction procedure and sharp peaks. Separations were performed on a new low hydrophobic weak cation-exchange analytical column. This technique is simple, rapid and useful for routine checks in repetitive analyses. PMID:15065779

Cinquina, A L; Calì, A; Longo, F; De Santis, L; Severoni, A; Abballe, F



Effect of modulator sorption on gradient shape in ion-exchange chromatography  

NASA Technical Reports Server (NTRS)

Mobile phase additives, or modulators, are used in gradient elution chromatography to facilitate separation and reduce separation time. The modulators are usually assumed to be linearly adsorbed or unadsorbed. Here, the consequences of nonlinear modulator adsorption are examined for ion-exchange gradient elution through a series of simulations. Even when the buffer salt is identical to the modulator salt, gradient deformation is observed; the extent of deformation increases as the volume of the feed is increased. When the modulator salt is different from the buffer salt, unusual effects are observed, and the chromatograms are quite different from those predicted by classical gradient elution theory. In particular, local increases in the buffer concentration are found between feed bands, and serve to improve the separation. These effects become more pronounced as the feed volume increases, and could therefore prove valuable in preparative applications.

Velayudhan, A.; Ladisch, M. R.; Mitchell, C. A. (Principal Investigator)



Separation selectivity of some ethylenediaminetetraacetic acid and cyclohexane-1,2-diaminetetraacetic acid complexes in column and ion electrokinetic chromatography.  


The complexes of Mn2+, Cd2+, Fe3+, Pb2+, Ni2+, Co2+, Zn2+ and Cu2+ with EDTA and cyclohexane-1,2-diaminetetraacetic acid (CDTA) were separated and detected in column and ion electrokinetic chromatography with suppressed conductivity and direct UV detection, respectively. In column ion chromatography (IC) these complexes were separated on an IonPac AS4A anion-exchange column (Dionex, USA). Parameters of carrier electrolyte, which were examined in the ion electrokinetic chromatography (IEKC) mode, include polymer and sulfate concentrations. In IEKC separation selectivity of complexes with poly(diallyldimethylammonium) cation as modifier is similar as for an IonPac AS4A column both for EDTA and CDTA chelates. It was shown that the ion-exchange capacity of the electrokinetic system is more than 100-times lower than the capacity of the IC column for the same peak resolution. In comparison with column main advantages of electrokinetic version are high separation efficiency (220,000-390,000 theoretical plates) and the absence of the analyte interaction with the sorbent matrix. PMID:10457487

Krokhin, O V; Adamov, A V; Hoshino, H; Shpigun, O A; Yotsuyanagi, T



Chemical Speciation Analysis of Sports Drinks by Acid-Base Titrimetry and Ion Chromatography: A Challenging Beverage Formulation Project  

ERIC Educational Resources Information Center

Students have standardized a sodium hydroxide solution and analyzed commercially available sports drinks by titrimetric analysis of the triprotic citric acid, dihydrogen phosphate, and dihydrogen citrate and by ion chromatography for chloride, total phosphate and citrate. These experiments are interesting examples of analyzing real-world food and…

Drossman, Howard




EPA Science Inventory

Bromate is a disinfection byproduct in drinking water which is formed during the ozonation of source water containing bromide. This paper described the analysis of bromate via ion chromatography-inductively coupled plasma mass spectrometry. The separation of bromate from interfer...


Determination of total sulfur by ion chromatography following peroxide oxidation in spent caustic from the chemical cleaning of coal  

Microsoft Academic Search

Total sulfur in samples of spent caustic arising from the chemical cleaning of coal has been determined by ion chromatography after oxidation of all sulfur species to sulfate. Oxidation with hydrogen peroxide first under basic conditions and subsequently under strongly acidic conditions was required for quantitative conversion of all sulfur species to sulfate. The effects of pH, sample size, and

Colin D. Chriswell; David R. Mroch; Richard. Markuszewski



Scaled-up separation of cellobiohydrolase1 from a cellulase mixture by ion-exchange chromatography.  


Enzymatic hydrolysis of cellulose often involves cellulases produced by Trichoderma reesei, of which cellobiohydrolase1 (CBH1) is the most abundant (about 60% of total cellulases) and plays an important role in the hydrolysis of crystalline cellulose. A method for separating sufficient quantities from the bulk cellulase cocktail is highly desirable for many studies, such as those that aim to characterize binding and hydrolysis kinetics of CBH1. In this work, CBH1 was separated from other Spezyme CP cellulases by ion-exchange chromatography using an efficient modification of a smaller scale process. The ion-exchange column was connected to a vacuum manifold system to provide a steady flow through parallel columns and thus achieve scale-up for enzyme separation. With five 5-mL columns running in parallel, about 55 mg of CBH1 was separated from 145 mg of Spezyme CP in a single separation. Step elution was used to replace the continuous gradient used at smaller scale. The purified CBH1 was collected in the fraction eluted with a buffer containing 0.33 M salt and showed comparable purity and activity as the enzyme purified by a fast protein liquid chromatography system. The stability of separated CBH1 was studied for up to 2 days and good thermal stability was observed. Separated CBH1 also showed both high adsorption to bacterial microcrystalline cellulose with ~4 ?mol/g maximum adsorption and a K(a) of 5.55 ± 2.34 ?M(-1) , and good hydrolytic activity based on atomic force microscopy observations that show a reduction in fiber height. PMID:21905272

Ye, Zhuoliang; Lane, Andrew N; Willing, Gerold A; Berson, R Eric



Formation of iron complexs from trifluoroacetic acid based liquid chromatography mobile phases as interference ions in liquid chromatography/electrospray ionization mass spectrometric analysis  

SciTech Connect

Two unexpected singly charged ions at m/z 1103 and 944 have been observed in mass spectra obtained from electrospray ionization-mass spectrometric analysis of liquid chromatography effluents with mobile phases containing trifluoroacetic acid. Accurate mass measurement and tandem mass spectrometry studies revealed that these two ions are not due to any contamination from solvents and chemicals used for mobile and stationary phases or from the laboratory atmospheric environment. Instead these ions are clusters of trifluoroacetic acid formed in association with acetonitrile, water and iron from the stainless steel union used to connect the column with the electrospray tip and to apply high voltage; the molecular formulae are Fe+((OH)(H2O)2)9(CF3COOH)5 and Fe+((OH)(H2O)2)6 (CF3COOH)5.

Shukla, Anil K.; Zhang, Rui; Orton, Daniel J.; Zhao, Rui; Clauss, Therese RW; Moore, Ronald J.; Smith, Richard D.



Ion-pair reversed-phase liquid chromatography for determination of polar underivatized amino acids using perfluorinated carboxylic acids as ion pairing agent  

Microsoft Academic Search

An isocratic liquid chromatography method using volatile ion pairing agent and evaporative light scattering detection was developed for the analysis of 10 underivatized amino acids (Asp, Asn, Ser, Gly, Gln, Cys, Glu, Thr, Ala, Pro) which are not sufficiently retained and resolved in the LC system commonly used. Five perfluorinated carboxylic acids (trifluoroacetic, heptafluorobutyric, nonafluoropentanoic, tridecafluoroheptanoic and pentadecafluorooctanoic acid) have

K. N Petritis; P Chaimbault; C Elfakir; M Dreux



Ambient temperature nanoelectrospray ion mobility detector for high performance liquid chromatography in determining amines.  


A nanoelectrospray ionization ion mobility spectrometer (nanoESI-IMS) working at ambient pressure and ambient temperature was developed as a detector of high performance liquid chromatography (HPLC) to achieve sensitive detection of amines with no derivatization and meanwhile provide another dimension of separation. The easier desolvation property of the charged droplets formed in nanoESI source enabled complete desolvation of the product ions of sixteen amines and drugs using the nanoESI-IMS at ambient temperature. Working at ambient temperature was good for suppressing the dissociation of thermal volatile ions, such as only the proton adducted molecular ions were observed for morphine in the nanoESI-IMS. Besides, the resolving power of the nanoESI-IMS also showed an increasing tendency as lowering the working temperature, an increment of 19 percent and 10 percent was observed for diethylamine and triethylamine as the temperature dropped from 92°C to 32°C. The resolving power of the nanoESI-IMS at 32°C for the 16 tested compounds was amid 33-44. With the nanoESI-IMS coupled to HPLC, a six-compound mixture including isomers was successfully separated and detected without any derivatization. And linear response ranges of 1 to 20, 0.5 to 20, and 0.8 to 20?gml(-1) and limits of detection of 0.25, 0.15, and 0.17?gml(-1) for triethylamine, diethylamine, and butylamine, respectively, were obtained with the hyphenated system. These results showed the excellent performance of the two-dimensional separation and detection method in direct qualitative and quantitative analyses of amines. PMID:25086751

Chen, Chuang; Hou, Keyong; Wang, Weiguo; Li, Jinghua; Li, Haiyang



Continuous processing of recombinant proteins: Integration of inclusion body solubilization and refolding using simulated moving bed size exclusion chromatography with buffer recycling.  


An integrated process which combines continuous inclusion body dissolution with NaOH and continuous matrix-assisted refolding based on closed-loop simulated moving bed size exclusion chromatography was designed and experimentally evaluated at laboratory scale. Inclusion bodies from N(pro) fusion pep6His and N(pro) fusion MCP1 from high cell density fermentation were continuously dissolved with NaOH, filtered and mixed with concentrated refolding buffer prior to refolding by size exclusion chromatography (SEC). This process enabled an isocratic operation of the simulated moving bed (SMB) system with a closed-loop set-up with refolding buffer as the desorbent buffer and buffer recycling by concentrating the raffinate using tangential flow filtration. With this continuous refolding process, we increased the refolding and cleavage yield of both model proteins by 10% compared to batch dilution refolding. Furthermore, more than 99% of the refolding buffer of the raffinate could be recycled which reduced the buffer consumption significantly. Based on the actual refolding data, we compared throughput, productivity, and buffer consumption between two batch dilution refolding processes - one using urea for IB dissolution, the other one using NaOH for IB dissolution - and our continuous refolding process. The higher complexity of the continuous refolding process was rewarded with higher throughput and productivity as well as significantly lower buffer consumption compared to the batch dilution refolding processes. PMID:24169042

Wellhoefer, Martin; Sprinzl, Wolfgang; Hahn, Rainer; Jungbauer, Alois



Combining size-exclusion chromatography and fully automated chip-based nanoelectrospray quadrupole time-of-flight tandem mass spectrometry for structural analysis of chondroitin/dermatan sulfate in human decorin.  


Chondroitin/dermatan sulfate (CS/DS) chain of decorin (DCN) from human skin fibroblasts (HSk) was released by reductive ?-elimination reaction and digested with chondroitin AC I lyase. Enzymatic hydrolysis mixture of CS/DS chains was separated by size-exclusion chromatography (SEC). Collected octasaccharide fraction was subjected to fully automated chip-based nanoelectrospray (nanoESI) quadrupole time-of-flight (QTOF) MS and tandem MS (MS/MS). MS of human skin fibroblasts DCN CS/DS displayed a high complexity due to the large variety of glycoforms, which under chip-nanoESI MS readily ionized to form multiply charged ions. Except for the regularly tetrasulfated octasaccharide, the investigated fraction contained four additional octasaccharides of atypical sulfation status. Two new oversulfated glycoforms and two undersulfated species were identified. Remarkably, the series of decasaccharides discovered in the same SEC pool was found to encompass a trisulfated and a novel hexasulfated [4,5-?-GlcAGalNAc(IdoAGalNAc)?] species. MS/MS by collision-induced dissociation (CID) on the [M-4H]? ion corresponding to the previously not reported [4,5-?-GlcAGalNAc(IdoAGalNAc)?](5S) corroborated for a novel motif in which three N-acetylgalactosamine (GalNAc) moieties are monosulfated, 4,5-?-GlcA and the first IdoA from the non-reducing end bear one sulfate group each, while the second N-acetylgalactosamine from the reducing end is unsulfated. PMID:21647927

Zamfir, Alina D; Flangea, Corina; Sisu, Eugen; Seidler, Daniela G; Peter-Katalini?, Jasna



The use of on-line ion chromatography for high temperature and high pressure reaction studies  

SciTech Connect

This paper describes the use of on-line ion chromatography as a tool for chemistry reaction studies in small volume systems. The technique was used to study chemistry behavior in a high temperature and high pressure autoclave system. A dual analyzer, multi-channel on-line ion chromatograph (IC) was configured to automate the sampling and analysis. Analytical channels were set up for analysis of inorganic anions, monovalent cations, conductivity, and pH. Conductivity and pH were measured using the IC as a flow injection analyzer. Use of the IC system provides significant advantages over conventional sampling and analysis techniques: Reduction in sample volume, a closed sampling system that protects air or light sensitive analytes from breakdown, around-the-clock test performance combined with automatic calibration and quality control checking, and detection and tracking of reaction products or unexpected contaminants. Methods used to correct measured concentrations for the effects of sampling and for calculation of control chemical loss half-lives are presented. A limited evaluation of the flow injection analysis methods for conductivity and pH is provided.

Lynch, G.J.



Design for gas chromatography-corona discharge-ion mobility spectrometry.  


A corona discharge ionization-ion mobility spectrometry (CD-IMS) with a novel sample inlet system was designed and constructed as a detector for capillary gas chromatography. In this design, a hollow needle was used instead of a solid needle which is commonly used for corona discharge creation, helping us to have direct axial interfacing for GC-IMS. The capillary column was passed through the needle, resulting in a reaction of effluents with reactant ions on the upstream side of the corona discharge ionization source. Using this sample introduction design, higher ionization efficiency was achieved relative to the entrance direction through the side of the drift tube. In addition, the volume of the ionization region was reduced to minimize the resistance time of compounds in the ionization source, increasing chromatographic resolution of the instrument. The effects of various parameters such as drift gas flow, makeup gas flow, and column tip position inside the needle were investigated. The designed instrument was exhaustively validated in terms of sensitivity, resolution, and reproducibility by analyzing the standard solutions of methyl isobutyl ketone, heptanone, nonanone, and acetophenone as the test compounds. The results obtained by CD-IMS detector were compared with those of the flame ionization detector, which revealed the capability of the proposed GC-IMS for two-dimensional separation (based on the retention time and drift time information) and identification of an analyte in complex matrixes. PMID:23083064

Jafari, Mohammad T; Saraji, Mohammad; Sherafatmand, Hossein



Separation of amaranthine-type betacyanins by ion-pair high-speed countercurrent chromatography.  


Betacyanins, red-violet plant pigments, were fractionated by ion-pair high-speed countercurrent chromatography (IP-HSCCC) from leaves extract of Iresine lindenii Van Houtte, an ornamental plant of the family Amaranthaceae. An HSCCC solvent system consisting of TBME-1-BuOH-ACN-H2O (1:3:1:5, v/v/v/v) was applied using ion-pair forming heptafluorobutyric acid (HFBA). Significantly different elution profiles of betacyanin diastereomeric pairs (derivatives based on betanidin and isobetanidin) observed in the HSCCC in comparison to HPLC systems indicate a complementarity of both techniques' fractionation capabilities. The numerous diastereomeric pairs can be selectively separated from each other using the HSCCC system simplifying the pigment purification process. Apart from the three well known highly abundant pigments (amaranthine, betanin and iresinin I) together with their isoforms, three new acylated (feruloylated and sinapoylated) betacyanins as well as known pigment hylocerenin (previously isolated from cacti fruits) were characterized in the plant for the first time and they are new for the whole Amaranthaceae family. PMID:24767836

Jerz, Gerold; Gebers, Nadine; Szot, Dominika; Szaleniec, Maciej; Winterhalter, Peter; Wybraniec, Slawomir



Collection of ethanolamines in air and determination by mobile phase ion chromatography  

SciTech Connect

A method is described for the collection and determination of monoethanolamine (MEA), diethanolamine (DEA) and triethanolamine (TEA) in air. Samples were collected by pulling air through a glass tube containing alumina, cleaned especially to remove interfering inorganic ions. The ethanolamines were desorbed with water and determined by Mobile Phase Ion Chromatography (MPIC). The recovery and total relative precision for MEA, DEA, and TEA - all collected from air at a flow rate of 100 mL/min for 7 hr - was 93.1 +/- 17%, 92.7 +/- 15% and 89.4 +/- 21%, respectively (95% confidence level). The method was validated for all three compounds from approximately the limit of detection (3 x noise) to ten times the limit of detection. Based on a sample size of 42 L, MEA was validated over the range from 0.12 to 3.0 ppm v/v (TLV=3), DEA over the range from 0.25 to 3.3 ppm v/v (TLV=3) and TEA from 0.31 to 3.7 ppm v/v (no TLV assigned). No effect on recovery was observed when sampling at high humidity or on storage of the samples for up to 31 days.

Bouyoucos, S.A.; Melcher, R.G.



Chromatography Theory  

NSDL National Science Digital Library

This site contains standard definitions related to chromatography similar to treatments found in analytical chemistry textbooks. It introduces the beginning student to Liquid Chromatography concepts relevant to biochemistry and includes a good example of choosing a mobile phase pH for a protein separation based on ion exchange.


Ion chromatography to detect salts in stone structures and to assess salt removal methods  

NASA Astrophysics Data System (ADS)

Stone - and in general all materials- from built heritage is very often damaged by salt crystallisation processes. Such processes usually derive into a loss of material compactness, as salts - given specific conditions and parameters- crystallize inside the material pores, exerting a pressure against the material pore walls higher than what they can resist - similar to the effect of liquid water when converts to solid water or ice-, thus breaking and disrupting the material by generating fissures and increasing the pore volume ratio, loosing its initial cohesion. When these deterioration processes take place inside a structure, salts - from different sources: material itself, restoration materials, from the ground, etc.- may come up to the stone surface - either temporarily or in permanently-, from beneath it, as efflorescences, depending mainly on the microclimatic conditions of the environment and the salts source. Efflorescences can be analysed and their nature identified (e.g. by means of X ray diffraction, in which the mineralogical composition of the salt is obtained), which can be, general, of aid not only for restoration but for preventive conservation measures. But what we do not know a priori when only characterising salt compounds- is the extent of the damage due to the presence of salts inside a structure (sub- and cryptoefflorescences). In this work we present a procedure in which the depth of the salt content can be measured, and its nature identified, based on the use of the ion chromatography technique. This technique allows identifying the existing ions in a specific sample, both anions and cations. The procedure consists of drilling (with a drilling core ranging from 5 to 8 mm in diameter, therefore causing the minimum damage to the material) in a same point at different depths from the surface and several depths from the bottom. The samples obtained are analysed and the ion content determined, qualitative and quantitatively. By means of a thorough previous inspection, we can select the most representative points by a drilling net - as minimum as possible- and make some profiles of the inner salt content of a structure. Moreover, this procedure is not only reliable for determining the nature and extent of salts damage, but to assess the efficacy of salts removal methods in cultural heritage. Here we present two case studies from relevant buildings of the Spanish cultural heritage in which this procedure was performed with successful and useful results, in both terms of understanding what types of salts were decaying the stones structures, and also whether the salts removal methods that were planned in the restoration project were efficient or not. It should be remarked that even ion chromatography is not a non destructive technique (can be considered as a minimally destructive one due to the few quantity it is needed for the analysis), the information it can provide is so useful that should not be ruled out from the beginning, depending of each specific case.

Alvarez de Buergo, M.; Lopez-Arce, P.; Fort, R.



Purification of RanGDP, RanGTP, and RanGMPPNP by ion exchange chromatography.  


Ran is a small GTPase that cycles between a guanosine diphosphate (GDP)-bound form (RanGDP) and a guanosine triphosphate (GTP)-bound form (RanGTP) and plays important roles in nuclear transport and mitosis. For studies of Ran function and its interactions with partner proteins, pure RanGDP and RanGTP complexes are critical. Ran complexed with the nonhydrolyzable GTP analog, GMPPNP (RanGMPPNP), is used instead of RanGTP when inhibition of hydrolysis is required. In this study, we demonstrate that the binding of Ran to a UNO Q ion exchange column is remarkably sensitive to small shifts in MgCl(2) concentration, and we use this property to purify recombinant RanGTP, RanGMPPNP, and RanGDP complexes. At 10 mM MgCl(2), Ran was found predominantly in the flow-through and, thus, was separated from the vast majority of bacterial proteins. After reducing the concentration of MgCl(2) to 5 mM, further purification of RanGTP, RanGMPPNP, and RanGDP was achieved by loading onto ion exchange columns and elution with an NaCl gradient. Purity of the resulting preparations was confirmed by releasing the bound nucleotide and checking it against a known nucleotide by high-performance liquid chromatography (HPLC). To further confirm the purity and function of the Ran preparations, appropriate protein-binding, enzymatic, and nuclear import assays were carried out. These methods should facilitate studies of cellular processes involving Ran by providing pure functional Ran-nucleotide complexes. PMID:15351280

Bibak, Niloufar; Paul, Rachelle M J; Freymann, Douglas M; Yaseen, Nabeel R



Determination of hexavalent chromium at the level of the California Public Health Goal by ion chromatography.  


Chromium is a primary drinking water contaminant in the USA with hexavalent chromium, Cr(VI), being the most toxic form of the metal. As a required step in developing a revised state drinking water standard for chromium, the California Department of Health Services recently issued a new Public Health Goal (PHG) of 2.5 microg/l for total chromium and 0.2 microg/l for Cr(VI). Hexavalent chromium can be determined (as chromate) by ion chromatography, as described in US Evironmental Protection Agency Method 218.6; however, the method as originally published does not allow sufficient sensitivity for analysis at the California PHG level of 0.2 microg/l. Modification of the conditions described in Method 218.6, including the use of a lower eluent flow-rate, larger reaction coil, and larger injection volume, significantly increases the method sensitivity. The modified method, which uses IonPac NG1 and AS7 guard and analytical columns, an eluent of 250 mM ammonium sulfate-100 mM ammonium hydroxide operated at 1.0 ml/min, a 1000 microl injection volume, and postcolumn reaction with 2 mM diphenylcarbazide-10% methanol-0.5 M sulfuric acid (using a 750 microl reaction coil) followed by UV-Vis detection at 530 nm, permits a method detection limit for chromate of 0.02 microg/l. This results in a quantitation limit of 0.06 microg/l, which is more than sufficient for analysis at the California PHG level. Calibration is linear over the range of 0.1-10 microg/l and quantitative recoveries (>80%) are obtained for chromate spiked at 0.2 microg/l in drinking water. The modified method provides acceptable performance, in terms of chromate peak shape and recovery, in the presence of up to 1000 mg/l chloride or 2000 mg/l sulfate. PMID:12108659

Thomas, D H; Rohrer, J S; Jackson, P E; Pak, T; Scott, J N



Separation of uremic toxins from urine with resorcinarene-based ion chromatography columns.  


People with chronic kidney disease suffer from uremic toxins which accumulate in their bodies. Detection and quantification of uremic toxins help diagnose kidney problems and start patient care. The aim of this research was to seek a new method to assist this diagnosis by trace level detection and separation of guanidine containing uremic toxins in water and urine. To detect and quantify the uremic toxins, new stationary phases for ion chromatography (IC) columns based on glutamic acid functionalized resorcinarenes bound to divinylbenzene macroporous resin were prepared. The new column packing material afforded separation of the five compounds: guanidinoacetic acid, guanidine, methylguanidine, creatinine, and guanidinobenzoic acid in 30min. Peak resolutions ranged from 7.6 to 1.3. Gradient elutions at ambient temperature with methanesulfonic acid (MSA) solution as eluent resulted in detection levels in water from 10 to 47ppb and in synthetic urine from 28 to 180ppb. Limits of quantification for the analytes using pulsed amperometric detection were 30-160ppb in water and 93-590ppb in urine. Trace levels of creatinine (1ppm) were detected in the urine of a healthy individual using the columns. PMID:25537175

Panahi, Tayyebeh; Weaver, Douglas J; Lamb, John D; Harrison, Roger G



Fabrication of electrolytic cell for online post-column electrochemical derivatization in ion chromatography.  


An electrolytic cell (EC), composed of two ruthenium-plated titanium electrodes separated by cation-exchange membranes, was fabricated and evaluated for online postcolumn derivatization in ion chromatography (IC). Folic acid (FA) and methotrexate (MTX) were preliminarily used as prototype analytes to test the performance of EC. After separation by an anion exchange column, FA and MTX, which emit very weak fluorescence when excited, were electrochemically oxidized online in the anode chamber of the EC. The compounds with strong fluorescence, which are oxidation products, were detected by the fluorescence detector. The phosphate buffer solution (100 mM KH(2)PO(4)) served as an optimal eluent for anion exchange chromatographic separation and a suitable supporting electrolyte for electro-oxidation, leading to ideal compatibility between IC separation and the postcolumn electrochemical derivatization. For the presently proposed method, the linear ranges were from 0.01 mg L(-1) to 5 mg L(-1) for both FA and MTX. The detection limits of FA and MTX were 1.8 and 2.1 ?g L(-1), and the relative standard deviations (RSD, n=7) were 2.9% and 3.6%, respectively. The method was applied for the simultaneous determination of FA and MTX in the plasma of patients being treated for rheumatoid arthritis. The determination of MTX in the urine of the patients of diffuse large B cell lymphoma was also demonstrated. PMID:22713918

Wu, Shuchao; Xu, Wei; Yang, Bingcheng; Ye, Mingli; Zhang, Peimin; Shen-Tu, Chao; Zhu, Yan



Laboratory robotics -- An automated tool for preparing ion chromatography calibration standards  

SciTech Connect

This paper describes the use of a laboratory robot as an automated tool for preparing multi-level calibration standards for On-Line Ion Chromatography (IC) Systems. The robot is designed for preparation of up to six levels of standards, with each level containing up to eleven ionic species in aqueous solution. The robot is required to add the standards` constituents as both a liquid and solid additions and to keep a record of exactly what goes into making up every standard. Utilizing a laboratory robot to prepare calibration standards provides significant benefits to the testing environment. These benefits include: accurate and precise calibration standards in individually capped containers with preparation traceability; automated and unattended multi-specie preparation for both anion and cation analytical channels; the ability to free up a test operator from a repetitive routine and re-apply those efforts to test operations; The robot uses a single channel IC to analyze each prepared standard for specie content and concentration. Those results are later used as a measure of quality control. System requirements and configurations, robotic operations, manpower requirements, analytical verification, accuracy and precision of prepared solutions, and robotic downtime are discussed in detail.

Chadwick, J.L.




SciTech Connect

The Process Science Analytical Laboratory (PSAL) at the Savannah River National Laboratory was requested by the Defense Waste Processing Facility (DWPF) to develop and demonstrate an Ion Chromatography (IC) method for the analysis of glycolate, in addition to eight other anions (fluoride, formate, chloride, nitrite, nitrate, sulfate, oxalate and phosphate) in Sludge Receipt and Adjustment Tank (SRAT) and Slurry Mix Evaporator (SME) samples. The method will be used to analyze anions for samples generated from the Alternate Reductant Demonstrations to be performed for the DWPF at the Aiken County Technology Laboratory (ACTL). The method is specific to the characterization of anions in the simulant flowsheet work. Additional work will be needed for the analyses of anions in radiological samples by Analytical Development (AD) and DWPF. The documentation of the development and demonstration of the method fulfills the third requirement in the TTQAP, SRNL-RP-2010-00105, 'Task Technical and Quality Assurance Plan for Glycolic-Formic Acid Flowsheet Development, Definition and Demonstrations Tasks 1-3'.

Best, D.



A simple ion chromatography method for inorganic anion analysis in edible seaweeds.  


A new, simple, fast and sensitive ion chromatography (IC) method, for the simultaneous analysis of fluoride, chloride, nitrite, bromide, nitrate, phosphate and sulphate in edible seaweeds was developed and reported for the first time. The validation of the analytical method was studied in terms of linearity, sensitivity, precision and accuracy. All standard calibration curves showed very good correlation between anion peak area and concentration (r>0.999). Limits of detection and quantitation ranged between 0.002-0.05 mg/L and 0.01-0.1mg/L, respectively and indicated the high sensitivity of the method. Relative standard deviation values of repeatability and inter-day precision for standard anions with the same sample were less than 2%. Anion recoveries ranged from 97 to 113% for chloride and from 87 to 105% for sulphate, respectively and showed the fairly good accuracy of the method. The method was applied to the analysis of inorganic anions in brown and red edible seaweeds. Brown seaweeds were characterized by higher chloride content up to 33.7-36.9%, while red seaweeds were characterized by higher sulphate content (45-57%). Sulphate content in seaweeds is related to the presence of sulphated polysaccharides of biological importance. The method developed was well applicable to mineral anion analysis in edible seaweeds and shows suitability and reliability of use in other food samples of nutritional importance. PMID:20801334

Gómez-Ordóñez, Eva; Alonso, Esther; Rupérez, Pilar



Interfacing multistage mass spectrometry with liquid chromatography or ion mobility separation for synthetic polymer analysis.  


Synthetic polymers are naturally mixtures of homologs, even in pure form. More complexity is introduced by the presence of different comonomers, end groups and/or macromolecular architectures. The analysis of such systems is substantially facilitated by interfacing mass spectrometry (MS), which disperses based on mass, with an additional level of separation involving either interactive liquid chromatography (LC) or ion mobility (IM) spectrometry, both of which are readily coupled online with electrospray ionization and MS detection. IM-MS separates in the gas phase, post-ionization and, therefore, is ideally suitable for labile and reactive polymers. Its usefulness is illustrated with the characterization of non-covalent siloxane-saccharide complexes, metallosupramolecular assemblies and an air- and moisture-sensitive inorganic polymer, poly(dichlorophosphazene). Conversely, LC-MS which separates in solution phase, before ionization, is most effective for the analysis of polymeric mixtures whose components differ in polarity. Interactive LC conditions can be optimized to disperse by the content of hydrophobic units, as is demonstrated for amphiphilic polyether copolymers and sugar-based nonionic surfactant blends. Both LC-MS and IM-MS can be extended into a third dimension by tandem mass spectrometry (MS(2)) studies on select oligomers, in order to obtain insight into individual end groups and isomeric architectures, comonomer sequences and degree of substitution, for example, by hydrophobic functionalities. PMID:22641724

Scionti, Vincenzo; Katzenmeyer, Bryan C; Solak, Nilüfer; Li, Xiaopeng; Wesdemiotis, Chrys



Comparison of performance of INAA, RNAA and ion chromatography for the determination of individual lanthanides.  


Lanthanides represent an interesting group of elements which are steadily gaining importance in science and in industry. Literature data suggest that they may have a role in regulating cellular processes and also in agriculture enhancing plant growth. Only few instrumental methods like NAA, ICP-OES and ICP-MS have potential for the determination of low levels of all lanthanides, but in practice they often have to be combined with suitable separation/preconcentration methods to achieve maximum number of elements being determined and to assure good accuracy and precision. In this work an attempt has been made to compare the performance of both variants of NAA: purely instrumental activation analysis (INAA) and radiochemical mode (RNAA) with pre- and post-irradiation group separation for the determination of individual lanthanides with the special emphasis on rarely determined elements (Gd, Ho, Er, Tm, Lu). This has been done by the analysis of two reference materials with different matrices viz. biological (CTA-OTL-1) and mineral (CTA-FFA-1). The results are compared with those obtained by ion chromatography (IC) employing the same lanthanide group preseparation scheme as in the case of RNAA. PMID:19804983

Dybczy?ski, Rajmund S; Czerska, Elzbieta; Danko, Bozena; Kulisa, Krzysztof; Samczy?ski, Zbigniew



Quantification of four arsenic species in fruit juices by ion-chromatography–inductively coupled plasma–mass spectrometry  

Microsoft Academic Search

A method using ion chromatography–inductively coupled plasma–mass spectrometry (IC–ICP–MS) for the quantification of arsenic species in fruit juices has been developed and validated. The method is capable of quantifying four anionic arsenic species – arsenite (As(III)), arsenate (As(V)), dimethylarsinic acid (DMA) and monomethylarsonic acid (MMA) – in the presence of unretained species such as arsenobetaine (AsB). Method validation was based

Sean D. Conklin; Peter E. Chen



Characterisation of aquatic humic and non-humic matter with size-exclusion chromatography--organic carbon detection--organic nitrogen detection (LC-OCD-OND).  


Size-exclusion chromatography in combination with organic carbon detection (SEC-OCD) is an established method to separate the pool of NOM into major fractions of different sizes and chemical functions and to quantify these on the basis of organic carbon. One specific approach, also known as LC-OCD-OND, is based on the Gräntzel thin-film UV-reactor. This approach is described with recent improvements in fraction assignation (humic substances, biopolymers, building blocks, low molecular weight organic acids and neutrals, hydrophobic organic carbon), the coupling of a novel organic nitrogen detector (OND), and an improved diagram for the characterisation of aquatic humic substances (HS-diagram). The diagram replaces the operational distinction between humic and fulvic acids by a continuum ranging from aquagenic fulvic acids to pedogenic humic acids. PMID:20937513

Huber, Stefan A; Balz, Andreas; Abert, Michael; Pronk, Wouter



Characterization and quantitative amino acids analysis of analgesic peptides in cinobufacini injection by size exclusion chromatography, matrix-assisted laser desorption/ionization time of flight mass spectrometry and gas chromatography mass spectrometry.  


Cinobufacini injection that comes from the water extract of Bufo bufo gargarizans Cantor skin is widely used for cancer treatment in China. Peptide is one of its major types of constituents, however the biological effects and content of this injection are little reported. In present study, the analgesic effect of peptides was determined and evaluated by in-vivo models. To characterize and quantitatively analyze these peptides, a reliable and efficient method combining size exclusion chromatography and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry with amino acid analysis was developed. The peptides presented as a series of analogs with similar molecular weights mostly ranging from 2 to 8 kDa. The amino acid analysis by gas chromatography mass spectrometry (GC-MS) was developed to determine both free and combined amino acids (FAA and CAA) in cinobufacini injection. This method achieved good linearity (R(2) , 0.9909-0.9999) and low limit of detection and quantification. FAA and CAA samples were efficiently analyzed by modified Phenomenex EZ: faast procedure. For the sample analysis, the method showed good repeatability (relative standard deviation, RSD???10%). For most FAA and CAA the mean recoveries were >80% with RSD <10%. The GC-MS based method is useful for quality assurance of both FAA and CAA in cinobufacini injection. Copyright © 2014 John Wiley & Sons, Ltd. PMID:24924921

Wu, Xu; Si, Nan; Bo, Gao; Hu, Hao; Yang, Jian; Bian, Baolin; Zhao, Hai Yu; Wang, Hongjie




Technology Transfer Automated Retrieval System (TEKTRAN)

Eleven B-lactams antibiotics were analyzed in fortified and incurred beef kidney tissue using high-performance liquid chromatography/selective reaction monitoring/tandem ion trap mass spectrometry. The analytes included: deacetylcephapirin, amoxicillin, cephapirin, desfuroylceftiofur cysteine disul...



EPA Science Inventory

This method is applicable to the identification and quantitation of perchlorate in raw and finished drinking waters. The approach used is ion chromatography with suppressed conductivity and electrospray ionization mass spectrometry (IC-ESI/MS)...



EPA Science Inventory

In this presentation the analytical instrumentation and procedures necessary to qualitatively and quantitatively determine low levels of perchlorate (ClO4-) in drinking waters using ion chromatography with electrolytic conductivity suppression, electrospray ionization mass spec...



Technology Transfer Automated Retrieval System (TEKTRAN)

A high performance size exclusion chromatographic method utilizing an evaporative light scattering detector was developed to separate and quantify galacturonic acid (GA) oligomers. Values of k for GA monomer ranged from 0.16 in water to 0.67 in 100 mM acetic acid. In 40 mM acetic acid calibration ...


Separation of betalains from berries of Phytolacca americana by ion-pair high-speed counter-current chromatography  

Microsoft Academic Search

The first preparative fractionation of betalain pigments by means of ion-pair high-speed counter-current chromatography (IP-HSCCC) from berry extracts of Phytolacca americana (Phytolaccaceae) is presented. A novel HSCCC solvent system consisting of 1-butanol–acetonitrile–water (5:1:6, v\\/v\\/v) was applied using ion-pair forming trifluoroacetic acid at low concentration (0.7%, v\\/v). Affinity of polar betacyanins and betaxanthins to the organic stationary phase of the biphasic

Gerold Jerz; Tanja Skotzki; Kathrin Fiege; Peter Winterhalter; S?awomir Wybraniec



A Volatile Organic Analyzer for Space Station - Description and evaluation of a gas chromatography/ion mobility spectrometer  

NASA Technical Reports Server (NTRS)

An on-board Volatile Organic Analyzer (VOA), an essential component of the Environmental Health System (EHS) air-quality monitoring strategy, is described. The strategy is aimed at warning the crew and ground personnel if volatile compounds exceed safe exposure limits. The VOA uses a combination of gas chromatography (GC) and ion-mobility spectrometry (IMS) for environmental monitoring and analysis. It is concluded that the VOA dual-mode detection capability and the ion mobilities in the drift region are unique features that can assist in the resolution of coeluting GC peaks. The VOA is capable of accurately identifying and quantifying target compounds in a complex mixture.

Limero, Thomas; Brokenshire, John; Cumming, Colin; Overton, ED; Carney, Ken; Cross, Jay; Eiceman, Gary; James, John



Analysis of kerogens by pyrolysis-gas chromatography-mass spectrometry using selective ion monitoring. III. Long-chain alkylbenzenes  

NASA Astrophysics Data System (ADS)

Analysis of the 3 principal kerogen types by pyrolysis-gas chromatography-mass spectrometry (P-GC-MS) has been undertaken using the mass spectrometer as a selective ion detector. By monitoring the fragment ions with {m}/{e} values of 91, 105 and 119 information has been obtained concerning differences in the content of long-chain alkylbenzenes in the kerogen pyrolysates, thus implying differences in the kerogen structures. Simulated maturation of the kerogens followed by specific ion ( {m}/{e} 105) gas chromatographic-mass spectrometric (GC-MS) analysis of the extracted liquid products, and specific ion ( {m}/{e} 105) GC-MS analysis of 3 crude oils is also described. The use of long-chain alkylbenzenes for geochemical correlation studies is discussed.

Solli, H.; Larter, S. R.; Douglas, A. G.


Fast and precise method for HPLC-size exclusion chromatography with UV and TOC (NDIR) detection: importance of multiple detectors to evaluate the characteristics of dissolved organic matter.  


A new type of high-performance liquid chromatography (HPLC)-size exclusion chromatography (SEC) system with ultraviolet (UV) absorbance detection and non-dispersive infrared (NDIR) detection of total organic carbon is described. The introduction of an online degassing tube and a low-volume HPLC column helped to reduce the analytical time and increase the sensitivity of the SEC system. This study is the first in which linear calibration curves (R(2)>0.99) were obtained for both UV absorbance and NDIR data for polystyrene sulfonate standards, which are the most suitable standards for molecular size analysis of aquatic humic substances as well as dissolved organic matter (DOM). Using the calibration curves, the molecular size distribution of DOM in water collected from Lake Kasumigaura and in pore water from lake sediments was estimated. Most of the DOM had a molecular weight less than 4000 Daltons (Da), and the amount of low-molecular-weight DOM (? 2000 Da) with low UV absorbance increased with depth in the sediment pore water. This result shows the importance of combining quantitative analysis by NDIR detection with qualitative analysis by UV detection to determine the chemical and physical properties of DOM. The possible sources and reactivity of DOM in Lake Kasumigaura and its sediment pore water are also discussed. PMID:21959091

Kawasaki, Nobuyuki; Matsushige, Kazuo; Komatsu, Kazuhiro; Kohzu, Ayato; Nara, Fumiko Watanabe; Ogishi, Fumikazu; Yahata, Masahito; Mikami, Hirohisa; Goto, Takeshi; Imai, Akio



Identification of conjugated linoleic acid isomers in cheese by gas chromatography, silver ion high performance liquid chromatography and mass spectral reconstructed ion profiles. Comparison of chromatographic elution sequences  

Microsoft Academic Search

Commercial cheese products were analyzed for their composition and content of conjugated linoleic acid (CLA) isomers. The\\u000a total lipids were extracted from cheese using petroleum ether\\/diethyl ether and methylated using NaOCH3. The fatty acid methyl esters (FAME) were separated by gas chromatography (GC), using a 100-m polar capillary column, into\\u000a nine minor peaks besides that of the major rumenic acid,

Najibullah Sehat; John K. G. Kramer; Magdi M. Mossoba; Martin P. Yurawecz; John A. G. Roach; Klaus Eulitz; Kim M. Morehouse; Youh Ku



Energy transduction in the thermophilic anaerobic bacterium Clostridium fervidus is exclusively coupled to sodium ions.  

PubMed Central

The thermophilic, peptidolytic, anaerobic bacterium Clostridium fervidus is unable to generate a pH gradient in the range of 5.5-8.0, which limits growth of the organism to a narrow pH range (6.3-7.7). A significant membrane potential (delta psi approximately -60 mV) and chemical gradient of Na+ (-Z delta pNa approximately -60 mV) are formed in the presence of metabolizable substrates. Energy-dependent Na+ efflux is inhibited by the Na+/H+ ionophore monensin but is stimulated by uncouplers, suggesting that the Na+ gradient is formed by a primary pumping mechanism rather than by secondary Na+/H+ antiport. This primary sodium pump was found to be an ATPase that has been characterized in inside-out membrane vesicles and in proteoliposomes in which solubilized ATPase was reconstituted. The enzyme is stimulated by Na+, resistant to vanadate, and sensitive to nitrate, which is indicative of an F/V-type Na(+)-ATPase. In the proteoliposomes Na+ accumulation depends on the presence of ATP, is inhibited by the ATPase inhibitor nitrate, and is completely prevented by the ionophore monensin but is stimulated by protonophores and valinomycin. These and previous observations, which indicated that secondary amino acid transport uses solely Na+ as coupling ion, demonstrate that energy transduction at the membrane in C. fervidus is exclusively dependent on a Na+ cycle. Images Fig. 2 PMID:8367451

Speelmans, G; Poolman, B; Abee, T; Konings, W N



Principles and Methods Chromatography  

E-print Network

Edition AC 18-1022-29 Principles and Methods Affinity Chromatography #12;Antibody Purification-1142-75 Protein Purification Handbook 18-1132-29 Ion Exchange Chromatography Principles and Methods 18-1114-21 Affinity Chromatography Principles and Methods 18-1022-29 Hydrophobic Interaction Chromatography Principles

Lebendiker, Mario


An interface for sensitive analysis of monoamine neurotransmitters by ion-pair chromatography-electrospray ionization-mass spectrometry with continuous online elimination of ion-pair reagents.  


A challenge in coupling ion-pair chromatography (IPC) online with electrospray ionization-mass spectrometry (ESI-MS) is that the nonvolatile ion-pair reagent (e.g., alkyl sulfate for amines or tetrabutylammonium for carboxylic acids) in the mobile phase suppresses the ESI-MS signals in the gas phase and their accumulation can clog the MS sampling interface. Consequently, IPC-ESI-MS is conducted either with a volatile ion-pair reagent, which could compromise the analyte separation efficiency, or with a downstream ion-exchange column to rid the ion-pair reagents of the mobile phase. In the latter approach, the limited capacity of ion-exchange columns requires frequent off-line column regeneration, which affects the separation throughput and prohibits long separations from being performed. A dual-valve, dual-ion exchange column interface of IPC-ESI-MS is designed for undisrupted separations and simultaneous column regeneration. Owing to the efficacy in removing the ion-pair reagent, the detection of eluents of monoamine neurotransmitters by an ion trap MS results in the limits of detection of 0.03 ?M for dopamine or DA and 0.01 ?M for 5-hydroxytryptamine or 5-HT. These values are lower than those obtained with ion trap MS of similar sensitivity when combined with the use of specialized chromatographic columns or sample preconcentration. Excellent reproducibility was attained with repeatedly regenerated ion-exchange columns (RSD = 4-6%) for an extended period of time (RSD < 6% for 6 days). DA and 5-HT in rat straital extracts were analyzed, and our data demonstrate that interferences inherent in the tissues and the ion-pair reagent have been successfully eliminated. This simple interface should be readily amenable to the separation and MS analysis of other types of polar compounds in complex sample media. PMID:23767971

Shi, Shuyun; Zhao, Binqing; Yagnik, Gargey; Zhou, Feimeng



[Analysis of piperidinium ionic liquid cations by ion chromatography with direct conductivity detection].  


A method of ion chromatography with direct conductivity detection was established to determine three piperidinium ionic liquid cations, i. e. N-methyl, ethyl piperidinium cation ([MEPi]+), N-methyl, propyl piperidinium cation ([MPPi]+) and N-methyl, butyl piperidinium cation ([MBPi]+). The effects of eluent types, eluent concentration and column temperature on the retention of the cations were investigated with sulfonic acid base cation exchange column using ethylenediamine-citric acid-acetonitrile as eluent. The results indicated that, with the increase of column temperature, the retention times of piperidinium cations were reduced, so the retention process of piperidinium cations was exothermic. The retentions of piperidinium cation homologue accorded with carbon number rule. The successful separation of the three piperidinium cations within 7 min was achieved using the optimized eluent of 0.2 mmol/L ethylenediamine-0.3 mmol/L citric acid-3% acetonitrile (pH 4.4) at a flow rate of 1.0 mL/min and the column temperature of 30 degrees C. Under these conditions, the detection limits at a signal-to-noise ratio of 3 for [MEPi]+, [MPPi] and [MBPi]+ were 0.14, 0.20 and 0.56 mg/L, respec-tively. The relative standard deviations (n = 5) for peak areas were less than 1.2%. The method has been applied to the determination of piperidinium ionic liquids synthesized in chemical laboratory with the spiked recoveries of 97.6% to 105.1%. The method is accurate, reliable, rapid, and has better practicability. PMID:23189670

Zhang, Renqing; Yu, Hong; Liu, Yuzhen



Purification of a recombinant baculovirus of Autographa californica M nucleopolyhedrovirus by ion exchange membrane chromatography.  


Significant progress in the application of viral vectors for gene delivery into mammalian cells and the use of viruses as biopesticides requires downstream processing that can satisfy application-specific demands on performance. In the present work the stability and ion exchange membrane chromatography of a recombinant of Autographa californica M nucleopolyhedrovirus is studied. To adjust the degree of purification the effect of ionic conductivity or pH on the viral infectivity was assessed (0.77-78.00mS/cm, pH 3-8). Infectivity decreased rapidly by several orders of magnitude at below 5mS/cm (i.e., 0.49MPa osmotic pressure change) or at below pH 5.5 (rationalized with particle aggregation). The virus was concentrated and purified via adsorption (0.2-1.1×10(16)pfu/m(3) chromatographic bed volume, 0.6-1.1×10(12)pfu/m(2) membrane area facing the incident fluid flow) and elution at pH 6.1 and 6.35mS/cm from three strong anion exchange membranes. Virus recovery and concentration in accord with the volume reduction were obtained using a polyether sulfone-based membrane with quaternary ammonium ligands. The level of host cell protein (down to below the detection limit) and suspended DNA (below 93pg DNA per 10(6)pfu) are reported for each membrane employed, for the purpose of comparability, under equal adsorption or elution conditions respectively. PMID:22521717

Grein, Tanja A; Michalsky, Ronald; Vega López, Maria; Czermak, Peter



A volatile organic analyzer for Space Station: Description and evaluation of a gas chromatography/ ion mobility  

NASA Technical Reports Server (NTRS)

A Volatile Organic Analyzer (VOA) is being developed as an essential component of the Space Station's Environmental Health System (EHS) air quality monitoring strategy to provide warning to the crew and ground personnel if volatile organic compounds exceed established exposure limits. The short duration of most Shuttle flights and the relative simplicity of the contaminant removal mechanism have lessened the concern about crew exposure to air contaminants on the Shuttle. However, the longer missions associated with the Space Station, the complex air revitalization system and the proposed number of experiments have led to a desire for real-time monitoring of the contaminants in the Space Station atmosphere. Achieving the performance requirements established for the VOA within the Space Station resource (e.g., power, weight) allocations led to a novel approach that joined a gas chromatograph (GC) to an ion mobility spectrometer (IMS). The authors of this paper will discuss the rational for selecting the GC/IMS technology as opposed to the more established gas chromatography/mass spectrometry (GC/MS) for the foundation of the VOA. The data presented from preliminary evaluations will demonstrate the versatile capability of the GC/IMS to analyze the major contaminants expected in the Space Station atmosphere. The favorable GC/IMS characteristics illustrated in this paper included excellent sensitivity, dual-mode operation for selective detection, and mobility drift times to distinguish co-eluting GC peaks. Preliminary studies have shown that the GC/IMS technology can meet surpass the performance requirements of the Space Station VOA.

Limero, Thomas F.; James, John T.



[Determination of sulfite in flue gas desulfurization with seawater by ion chromatography].  


The technology for flue gas desulfurization (FGD) with seawater is widely adopted by coal-fired power plants in coastal areas. SO2 in the flue gas is absorbed by alkaline seawater and transfered in aqueous phase as sulfite (SO3(2-)), and most SO3(2-) is transformed to sulfate (SO4(2-)) after an aeration process. The remaining SO3(2-) in the seawater discharged to sea area may be harmful to marine organism because of its biological toxicity, thus it is necessary to determine the concentration of SO3(2-) in the seawater for desulfurization. In this study, the method of determination of SO3(2-) in the seawater by ion chromatography was investigated. The separation was achieved on an IonPac AS14A column with 14 mmol/L NaOH-12 mmol/L Na2 CO3 solution as the mobile phase at a flow rate of 1.2 mL/min, and the detection was performed by a pulsed amperometric detector. Formaldehyde was added as a protective agent when sampling because the SO3(2-) is easy to be oxidized. To avoid the formation of Mg (OH)2 in the mobile phase with high pH value which might block the column, the Mg(2+) in seawater was precipitated by NaOH solution (pH 12.0) before sample determination. The method showed good linearity within the range of 0-100 mg/L with an average recovery of 116.8%. The method detection limit (MDL) reached as low as 0.05 mg/L. The relative standard deviations (RSD) for seawater matrix samples spiked at levels of 7.5, 25.0 and 75.0 mg/L were 2.1% 3.1% and 4.0% (n = 9) , respectively. The method has been applied for the determination of SO3(2-) in flue gas desulfurization seawater with the advantages of being fast, sensitive and selective. PMID:20352939

Yin, Liqian; Yuan, Dongxing; Guo, Juan; Liu, Xiyao



Method development for the redox speciation analysis of iron by ion chromatography-inductively coupled plasma mass spectrometry and carryover assessment using isotopically labeled analyte analogues.  


An ion chromatography-inductively coupled plasma mass spectrometry (IC-ICP-MS) method was developed for the redox speciation analysis of iron (Fe) based on in-column complexation of Fe(2+) and Fe(3+) by dipicolinic acid (DPA). The effects of column type, mobile phase composition and molecular ion interference were studied in the method optimization. The carryover of the target species in the IC-ICP-MS method was uniquely and effectively evaluated using isotopically enriched analogues of the analytes ((54)Fe(2+) and (57)Fe(3+)). Standard solutions of the enriched standards were injected into the system following analysis of a sample, and the ratios of the isotopes of iron in the enriched standards were calculated based on the chromatographic peak areas. The concentrations of the analytes carried over from the sample to the enriched standards were determined using the quantitative relationship in isotope dilution mass spectrometry (IDMS). In contrast to the routine way of evaluating carryover effect by injecting a blank solution after sample analysis, the use of isotopically enriched standards identified significant analyte carryover in the present method. Extensive experiments were carried out to systematically identify the source of the carryover and to eliminate the problem; the separation column was found to be the exclusive source. More than 95% of the analyte carryover was eliminated by reducing the length of the column. The detection limit of the IC-ICP-MS method (MDL) for the iron species was 2ngg(-1). The method was used to determine Fe(2+) and Fe(3+) in synthetic aqueous standard solutions and a beverage sample. PMID:24819017

Wolle, Mesay Mulugeta; Fahrenholz, Timothy; Rahman, G M Mizanur; Pamuku, Matt; Kingston, H M 'Skip'; Browne, Damien



An Ultra-Trace Analysis Technique for SF6 Using Gas Chromatography with Negative Ion Chemical Ionization Mass Spectrometry.  


Sulfur hexafluoride (SF6) is widely used as a tracer gas because of its detectability at low concentrations. This attribute of SF6 allows the quantification of both small-scale flows, such as leakage, and large-scale flows, such as atmospheric currents. SF6's high detection sensitivity also facilitates greater usage efficiency and lower operating cost for tracer deployments by reducing quantity requirements. The detectability of SF6 is produced by its high molecular electronegativity. This property provides a high potential for negative ion formation through electron capture thus naturally translating to selective detection using negative ion chemical ionization mass spectrometry (NCI-MS). This paper investigates the potential of using gas chromatography (GC) with NCI-MS for the detection of SF6. The experimental parameters for an ultra-trace SF6 detection method utilizing minimal customizations of the analytical instrument are detailed. A method for the detection of parts per trillion (ppt) level concentrations of SF6 for the purpose of underground ventilation tracer gas analysis was successfully developed in this study. The method utilized a Shimadzu gas chromatography with negative ion chemical ionization mass spectrometry system equipped with an Agilent J&W HP-porous layer open tubular column coated with an alumina oxide (Al2O3) S column. The method detection limit (MDL) analysis as defined by the Environmental Protection Agency of the tracer data showed the method MDL to be 5.2 ppt. PMID:25452581

Jong, Edmund C; Macek, Paul V; Perera, Inoka E; Luxbacher, Kray D; McNair, Harold M



Fast HPLC method using ion-pair and hydrophilic interaction liquid chromatography for determination of phenylephrine in pharmaceutical formulations.  


A rapid procedure based on a direct extraction and HPLC determination with fluorescence detection of phenylephrine in pharmaceutical sachets that include a large excess of paracetamol (65 + 1, w/w), ascorbic acid (5 + 1, w/w), and other excipients (aspartame and sucrose) was developed and validated. The final optimized chromatographic method for ion-pair chromatography used an XTerra RP18 column, 3 microm particle size, 50 x 3.0 mm id. The mobile phase consisted of a mixture of acetonitrile and buffer (10 mM sodium octane-1-sulfonate, adjusted with H3PO4 to pH 2.2; 200 + 800, v/v), with a constant flow rate of 0.3 mL/min. The separation was carried out at 30 degrees C, and the injection volume was 3 microL. Fluorescence detection was performed at excitation and emission wavelengths of 275 and 310 nm, respectively. The mobile phase parameters, such as the organic solvent fraction (acetonitrile) in mobile phase as an organic modifier, the concentration of sodium octane-1-sulfonate as a counter-ion, temperature, and pH of mobile phase, were studied. As an alternative to ion-pair chromatography, hydrophilic interaction liquid chromatography (HILIC) was investigated using a Luna HILIC column, 3 microm, 100 x 4.6 mm id. The mobile phase consisted of acetonitrile and buffer (5 mM potassium dihydrogen phosphate, adjusted with H3PO4 to pH 2.5; 750 + 250, v/v) at a flow rate of 0.8 mL/min. The separation was carried out at 25 degrees C, and the injection volume was 5 microL. The proposed method has an advantage of a very simple sample pretreatment, and is much faster than the currently utilized HPLC methods using gradient elution and UV detection. Commercial samples of sachets were successfully analyzed by the proposed HPLC method. PMID:21140654

Dousa, Michal; Gibala, Petr



Ion Chromatography Based Urine Amino Acid Profiling Applied for Diagnosis of Gastric Cancer  

PubMed Central

Aim. Amino acid metabolism in cancer patients differs from that in healthy people. In the study, we performed urine-free amino acid profile of gastric cancer at different stages and health subjects to explore potential biomarkers for diagnosing or screening gastric cancer. Methods. Forty three urine samples were collected from inpatients and healthy adults who were divided into 4 groups. Healthy adults were in group A (n = 15), early gastric cancer inpatients in group B (n = 7), and advanced gastric cancer inpatients in group C (n = 16); in addition, two healthy adults and three advanced gastric cancer inpatients were in group D (n = 5) to test models. We performed urine amino acids profile of each group by applying ion chromatography (IC) technique and analyzed urine amino acids according to chromatogram of amino acids standard solution. The data we obtained were processed with statistical analysis. A diagnostic model was constructed to discriminate gastric cancer from healthy individuals and another diagnostic model for clinical staging by principal component analysis. Differentiation performance was validated by the area under the curve (AUC) of receiver-operating characteristic (ROC) curves. Results. The urine-free amino acid profile of gastric cancer patients changed to a certain degree compared with that of healthy adults. Compared with healthy adult group, the levels of valine, isoleucine, and leucine increased (P < 0.05), but the levels of histidine and methionine decreased (P < 0.05), and aspartate decreased significantly (P < 0.01). The urine amino acid profile was also different between early and advanced gastric cancer groups. Compared with early gastric cancer, the levels of isoleucine and valine decreased in advanced gastric cancer (P < 0.05). A diagnosis model constructed for gastric cancer with AUC value of 0.936 tested by group D showed that 4 samples could coincide with it. Another diagnosis model for clinical staging with an AUC value of 0.902 tested by 3 advanced gastric cancer inpatients of group D showed that all could coincide with the model. Conclusions. The noticeable differences of urine-free amino acid profiles between gastric cancer patients and healthy adults indicate that such amino acids as valine, isoleucine, leucine, methionine, histidine and aspartate are important metabolites in cell multiplication and gene expression during tumor growth and metastatic process. The study suggests that urine-free amino acid profiling is of potential value for screening or diagnosing gastric cancer. PMID:22888338

Fan, Jing; Hong, Jing; Hu, Jun-Duo; Chen, Jin-Lian



Quantifying phytate in dairy digesta and feces: alkaline extraction and high-performance ion chromatography.  


Development of an analytical method with appropriate combination of extraction and quantification approaches for undigested phytate in ruminant feces and digesta will advance knowledge of phytate degradation in ruminants and help to reduce phosphorus excretion. Established quantification methods give satisfactory results for feedstuffs and nonruminant manure but recovery of phytate is incomplete for ruminant feces and digesta because of their complex sample matrix and low ratio of phytate to inorganic P. The objective was to develop a robust, accurate, sensitive, and inexpensive method to extract and quantify phytate in feeds, ruminant feces, and digesta. Diets varying in phytate content were fed to dairy heifers, dry cows, and lactating cows to generate digesta and fecal samples of varying composition to challenge extraction and quantification methods. Samples were extracted with 0.5 M HCl or 0.25 M NaOH + 0.05 M EDTA. Acid extracts were mixed with 20% NaCl, alkaline extracts were acidified to final pH < 2, and then both extracts were clarified with C?? cartridges and 0.2-?m filters. High-performance ion chromatography (HPIC) was used to quantify phytate. In feed samples, the measured phytate was comparable in alkaline and acid extracts (2,965 vs. 3,085 ?g/g of DM). In digesta and fecal samples, alkaline extraction yielded greater estimates of phytate content than did acid extraction (40.7 vs. 33.6 and 202.9 vs. 144.4 ?g/g of DM for digesta and fecal samples, respectively). Analysis of alkaline extracts by HPIC is usually not possible because of sample matrix interferences; acidification and C(18)-cartridge elution of alkaline extracts prevented this interference. Pure phytate added to dry samples before extraction was almost completely recovered (88 to 105%), indicating high extraction efficiency, no adverse effect of extract clean-up procedures, and accurate quantification of phytate. The proposed method is rapid, inexpensive, robust, and combines the extraction power of NaOH-EDTA with the precision and sensitivity of HPIC quantification, allowing accurate quantification of phytate in feeds, ruminant digesta, and fecal samples. PMID:22612959

Ray, P P; Shang, C; Maguire, R O; Knowlton, K F



Size exclusion chromatography for the unambiguous detection of aliphatics in fractions from petroleum vacuum residues, coal liquids, and standard materials, in the presence of aromatics  

SciTech Connect

A method has been developed using size exclusion chromatography (SEC) in heptane eluent that can detect aliphatics unambiguously without fractionation to remove aromatics. Spherical molecules such as colloidal silicas elute at the exclusion limit, while alkanes up to C{sub 50} elute through the porosity of the column. Detection of aliphatics was defined by use of an evaporative light scattering (ELS) detector with the simultaneous absence of UV absorbance at 300 nm. Alkanes smaller than C{sub 12} were not detected because the conditions of operation of the ELS caused their evaporation. All aromatics eluted after the permeation limit of about 25 min and were not detected until well after 45 min by their UV absorbance. The SEC method was applied to petroleum vacuum residues and coal liquids, and their fractions were soluble in pentane or heptane. High-temperature (HT) GC-MS confirmed the presence of alkanes in the pentane- and heptane-soluble fractions of petroleum vacuum residues, but did not elute any of the aromatics known to be present from SEC. Alkanes were examined in pentane-soluble fractions of a coal digest and a low-temperature coal tar; alkanes up to C{sub 40} were detected in the low-temperature tar and, although present in the digest, were masked by aromatics. No alkanes were detected by either SEC or HT GC-MS in fractions from a coal tar pitch. Aromatics in coal liquids and one petroleum residue were also examined by SEC using NMP as eluent and by UV fluorescence spectroscopy. The SEC method will find application to pentane- and heptane-soluble fractions of petroleum liquids and coal liquids where the alkanes are concentrated relative to the more abundant aromatics. 43 refs., 10 figs., 2 tabs.

Eiman M. Al-Muhareb; Fatma Karaca; Trevor J. Morgan; Alan A. Herod; Ian D. Bull; Rafael Kandiyoti [Imperial College, London (United Kingdom). Department of Chemical Engineering



An improved thin-layer chromatography/mass spectrometry coupling using a surface sampling probe electrospray ion trap system  

SciTech Connect

A combined surface sampling probe/electrospray emitter coupled with an ion trap mass spectrometer was used for the direct read out of unmodified reversed-phase C18 thin-layer chromatography (TLC) plates. The operation of the surface sampling electrospray ionization interface in positive and negative ionization modes was demonstrated through the direct analysis of TLC plates on which a commercial test mix comprised of four dye compounds viz., rhodamine B, fluorescein, naphthol blue black, and fast green FCF, and an extract of the caffeine-containing plant Ilex vomitoria, were spotted and developed. Acquisition of full-scan mass spectra and automated collection of MS/MS product ion spectra while scanning a development lane along the surface of a TLC plate demonstrated the advantages of using an ion trap in this combination. Details of the sampling system, benefits of analyzing a developed lane in both positive ion and negative ion modes, levels of detection while surface scanning, surface scan speed effects, and the utility of three-dimensional data display, are also discussed.

Ford, Michael J [ORNL; Van Berkel, Gary J [ORNL



Metabolomic profiling of anionic metabolites in head and neck cancer cells by capillary ion chromatography with Orbitrap mass spectrometry.  


A highly sensitive platform coupling capillary ion chromatography (Cap IC) with Q Exactive mass spectrometer has been developed for metabolic profiling of head and neck squamous cell carcinoma (HNSCC) cells. The Cap IC allowed an excellent separation of anionic polar metabolites, and the sensitivities increased by up to 100-fold compared to reversed-phase liquid chromatography and hydrophilic interaction chromatography performed at either high- or capillary-flow rates. The detection limits for a panel of standard metabolites were between 0.04 to 0.5 nmol/L (0.2 to 3.4 fmol) at a signal-to-noise ratio of 3. This platform was applied to an untargeted metabolomic analysis of head and neck cancer cells and stem-like cancer cells. Differential metabolomics analysis identified significant changes in energy metabolism pathways (e.g., glycolysis and tricarboxylic acid cycle). These experiments demonstrate Cap IC/MS as a powerful metabolomics tool by providing enhanced separation and sensitivity of polar metabolites combined with high resolution and accurate mass measurement (HR/AM) capabilities to differentiate isobaric metabolites. PMID:24766394

Wang, Junhua; Christison, Terri T; Misuno, Kaori; Lopez, Linda; Huhmer, Andreas F; Huang, Yingying; Hu, Shen



Absolute molecular weight determination of hypromellose acetate succinate by size exclusion chromatography: use of a multi angle laser light scattering detector and a mixed solvent.  


Molecular weight distribution is an important quality attribute for hypromellose acetate succinate (HPMCAS), a pharmaceutical excipient used in spray-dried dispersions. Our previous study showed that neither relative nor universal calibration method of size exclusion chromatography (SEC) works for HPMCAS polymers. We here report our effort to develop a SEC method using a mass sensitive multi angle laser light scattering detector (MALLS) to determine molecular weight distributions of HPMCAS polymers. A solvent screen study reveals that a mixed solvent (60:40%, v/v 50mM NaH(2)PO(4) with 0.1M NaNO(3) buffer: acetonitrile, pH* 8.0) is the best for HPMCAS-LF and MF sub-classes. Use of a mixed solvent creates a challenging condition for the method that uses refractive index detector. Therefore, we thoroughly evaluated the method performance and robustness. The mean weight average molecular weight of a polyethylene oxide standard has a 95% confidence interval of (28,443-28,793) g/mol vs. 28,700g/mol from the Certificate of Analysis. The relative standard deviations of average molecular weights for all polymers are 3-6%. These results and the Design of Experiments study demonstrate that the method is accurate and robust. PMID:21840663

Chen, Raymond; Ilasi, Nicholas; Sekulic, Sonja S



Assessment of the influence of amylose-LPC complexation on the extent of wheat starch digestibility by size-exclusion chromatography.  


Amylose forms inclusion complexes with lysophosphatidylcholine (LPC), that decrease the susceptibility of amylose to amylase degradation. This study on the influence of complexation on starch susceptibility to amylase explains the nature of this protective effect. Wheat starch suspensions (9% w/w) containing 0.5-5% LPC were subjected to hydrolysis by porcine pancreatic ?-amylase at 37 °C for several digestion times. The digesta were analysed by size-exclusion chromatography (SEC). The molar mass distribution was closely dependent on the digestion time and amount of LPC. This study precisely demonstrates the alteration of the digestion profile of starch on a molecular level, influenced by amylose-LPC complexation; however the effect depends on the digestion time. During 15 and 30 min digestion, inclusion complexes not only protect amylopectin in the initial hydrolysis stage, but also demonstrate lower susceptibility of the molecular amylose complexes to amylase hydrolysis. Digestion for 240 min resulted in a lower oligosaccharide peak concentration, in the presence of a high LPC concentration, which is related to less degradation of complexed amylose fraction. PMID:23993621

Ahmadi-Abhari, S; Woortman, A J J; Hamer, R J; Loos, K



Study of thermal aggregation of globulin from common buckwheat (Fagopyrum esculentum Moench) by size-exclusion chromatography and laser light scattering.  


The heat-induced aggregation of common buckwheat (Fagopyrum esculetum Moench) globulin (BWG) was studied using size-exclusion chromatography (SEC) combined with on-line multiangle laser light scattering (MALLS) and quasielastic light scattering (QELS). The unheated BWG was found to exist mainly as a hexamer, with an estimated weight-average molecular weight (M(w)) of 342 000, close to that deduced from the genomic cloned data of 13S buckwheat globulin. The QELS data predicted that the hexamer exists as two annular trimeric rings (diameter approximately 10.8 nm) placed on top of each other, forming an oblate cylinder (height approximately 9.1 nm). Upon heating, hexamers and trimers were dissociated and then associated to form extended small aggregates, finally forming compact, large macroaggregates. N-Ethylmaleimide would favor macroaggregate formation and increased the molar masses and hydrodynamic radii of the soluble aggregates, suggesting a different aggregation process in the presence of the sulfhydryl-blocking agent. A plot of log hydrodynamic radius versus log molar mass showed changes in the slope during heat treatment, suggesting conformational transformation in the heat-denatured and aggregated BWG molecules. PMID:16417320

Choi, Siu-Mei; Ma, Ching-Yung



Evaluations of the TiO2/simulated solar UV degradations of XAD fractions of natural organic matter from a bog lake using size-exclusion chromatography.  


This work reports on the changes in compositions of humic acids (HAs) and fulvic acids (FAs) during photocatalytic degradation. The HAs and FAs were obtained from the XAD-resin fractionation of natural-organic matter (NOM) from a bog lake (Lake Hohloh, Black Forest, Germany). Degussa P-25 titanium dioxide (TiO2) in a suspension and a solar UV simulator (batch reactor) were used in the experiments. The photocatalytic degradation of the HAs and FAs were monitored using size-exclusion chromatography (SEC) equipped with dissolved organic carbon (DOC) and ultraviolet (UV254) detection (SEC-DOC and SEC-UV254) and UV-Vis spectrophotometry. The evolutions of the photocatalytic degradations of the HA and FA fractions were selective. The photocatalytic degradation started with the degradations of high molecular weight compounds with relatively high UV254 absorbances in the HA and FA fractions to yield low molecular weight compounds showing less specific UV254 absorbances. Observance of the same tendency for the original NOM from Lake Hohloh indicates that these XAD-fractions still having complex compound mixtures. However, the larger molecular weight fractions of the FAs showed higher preferential adsorptions onto TiO2, which caused their faster degradation rates. Furthermore, FAs showed a greater reduction of the total THM formation potential (TTHMFP) and the organic halogen compounds adsorbable on activated carbon formation potential (AOXFP), in comparison with the HAs. PMID:23863374

Valencia, Sergio; Marín, Juan M; Restrepo, Gloria; Frimmel, Fritz H



Analysis of complex phthalic acid based polyesters by the combination of size exclusion chromatography and matrix-assisted laser desorption/ionization mass spectrometry.  


Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was used in conjunction with size exclusion chromatography (SEC) to investigate a model polyester system based on phthalic anhydride-1,2-propylene glycol. The polyesters were synthesized with a 30% molar excess of glycol, with kinetic samples being removed during different intervals of the polyesterification reaction. SEC was used to track the course of the reaction by determining the molecular weight and molecular weight distributions before subsequent off-line coupling with MALDI-TOF MS as a selective detection method to determine the chemical composition, identify the functionality type distributions as well as assist in assigning structural conformations. Mass spectrometry analysis proved to be a highly effective tool to facilitate the identification of the narrowly dispersed fractions obtained from the chromatographic separations as well as serve as a core method to investigate the heterogeneous nature of the bulk kinetic samples. Through the hyphenation of these sophisticated polymer characterization techniques, information on the molecular heterogeneity of the model polyesters, showing a complex variety of possible distributions, was obtained. PMID:24370096

Pretorius, Nadine O; Rode, Karsten; Simpson, Jaylin M; Pasch, Harald



Investigation of the elemental composition and chemical association of several elements in fulvic acids dietary supplements by size-exclusion chromatography UV inductively coupled plasma mass spectrometric.  


Four fulvic acid dietary supplement samples were obtained for this study with the intention of investigating the elemental composition and association of fulvic acids found in fulvic acid supplements. This was achieved by coupling size-exclusion chromatography (SEC) sequentially with UV-vis and inductively coupled plasma mass spectrometric (ICP-MS) detectors. The combination of UV and ICP-MS offered highly sensitive and selective detection. This technique was used in the present study to initially investigate the chemical association of several different elements including, Cr, Co, Ca, Fe, I, Mg, Zn, Se, Cu, Mn, Mo, As, Hg, Pb, and Ag, by observing the elution profile of the fulvic acids obtained with UV detection and matching their retention times with the peaks measured with ICP-MS. The results found based on this type of analysis suggest that there was some association of the elements to the fulvic acids. It was also of interest to observe the stability of these complexes upon human digestion; therefore a gastric digestion was mimicked. In the fulvic acid dietary supplement samples studied, fulvic acids were present in the samples and there was elemental association based on the retention time overlap in the UV as well as the ICP-MS. The fulvic complexes found in the samples were of a low molecular weight As a result of the digestion the SEC-ICP-MS chromatographic profile in some of the samples changed, which may infer that the elemental association had changed. PMID:15553158

Grant, Tyre D; Wuilloud, Rodolfo G; Wuilloud, Jorgelina C; Caruso, Joseph A



Impact of protein concentration on the determination of thiolic groups of ovalbumin: a size exclusion chromatography-chemical vapor generation-atomic fluorescence spectrometry study via mercury labeling.  


We optimized a hyphenated system based on size exclusion chromatography coupled to a microwave/UV mercury oxidation system and an atomic fluorescence detector (SEC-CVG-AFS) for the online oxidation of free and protein-complexed p-hydroxymercuribenzoic acid (pHMB) without the employment of chemical oxidizing agents. This system has been applied to the study of labeling of thiolic groups of native ovalbumin (OVA) as a function of protein concentration. We found that the protein concentration strongly affects the species distribution of OVA, the number of thiolic groups titrated in each species, and thus, the accuracy in the determination of the total number of thiolic groups. The amount of titrated sulfhydryl groups in the protein concentration range investigated (5-100 ?mol L(-1)) varied from 2.40 ± 0.01 to 1.85 ± 0.05 for the monomeric form of OVA and from 4.63 ± 0.01 to 5.63 ± 0.05 for the total OVA, which represents more than four theoretical number of reduced Cys. This information is important from the analytical point of view because it suggests that, unless to operate with diluted concentration of protein, the number of titrated thiolic groups results from both the aspecific interaction of the probe with aggregates species and to the specific bond of the probe with the accessible -SH groups. PMID:24502672

Campanella, Beatrice; Onor, Massimo; D'Ulivo, Alessandro; Giannarelli, Stefania; Bramanti, Emilia



Size exclusion and reversed-phase high-performance liquid chromatography/UV for routine control of thermal processing of cows' and donkey milk major proteins.  


Cows' and donkey milks (raw and thermally processed) and respective whey were analysed for quantification of major proteins. Two different chromatographic approaches, size exclusion (SE-HPLC) and reversed-phase high performance liquid chromatography (RP-HPLC) both coupled to UV detection were used. Usefulness of these methods for routine control of the effect of thermal processing was evaluated. The external standard method was used to calibrate the SE-HPLC and RP-HPLC systems. Concerning quantification of ?-lactoglobulin (?-lg), ?-lactalbumin (?-la), lysozyme (lys), and total casein (cn), no significant differences between results obtained by SE-HPLC and by RP-HPLC (t-test, P>0·05) were observed for raw milks and whey. Heating of cows' milk promoted aggregation of denatured proteins as observed by SE-HPLC, whereas ?-la and ?-lg from donkey milk were stable to thermal processing at 100 °C (5 min). Lys was quantified in donkey raw milk and whey however, in thermally processed donkey milk lys was denatured and could not be quantified by HPLC. PMID:22420770

Pinho, Carina; Martins, Zita E; Petisca, Catarina; Figurska, Agata M; Pinho, Olívia; Ferreira, Isabel M P L V O



Online Size-Exclusion High-Performance Liquid Chromatography Light Scattering and Differential Refractometry Methods to Determine Degree of Polymer Conjugation to Proteins and Protein–Protein or Protein–Ligand Association States  

Microsoft Academic Search

Characterizing the solution structure of protein–polymer conjugates and protein–ligand interactions is important in fields such as biotechnology and biochemistry. Size-exclusion high-performance liquid chromatography with online classical light scattering (LS), refractive index (RI), and UV detection offers a powerful tool in such characterization. Novel methods are presented utilizing LS, RI, and UV signals to rapidly determine the degree of conjugation and

Brent S. Kendrick; Bruce A. Kerwin; Byeong S. Chang; John S. Philo



The use of capillary viscometry, reducing end-group analysis, and size exclusion chromatography combined with multi-angle laser light scattering to characterize endo-1,4-?- d-glucanases on carboxymethylcellulose: a comparative evaluation of the three methods  

Microsoft Academic Search

Three methods were used to characterize seven purified endo-1,4-?-d-glucanases derived from different microbial sources (Trichoderma reesei, Thermomonospora fusca, and Acidothermus cellulolyticus) on carboxymethylcellulose (CMC). The methods included capillary viscometry, reducing end-group analysis, and high performance size exclusion chromatography combined with multi-angle laser light scattering (HPSEC-MALLS). The investigation was performed with the objective of comparative evaluation of the different methods for

Elena Yu Vlasenko; Anya I. Ryan; Charles F. Shoemaker; Sharon P. Shoemaker



Application of gas chromatography-mass spectrometry with selected ion monitoring to the urinanalysis of 4-pyridoxic acid.  


An analytical protocol has been developed for the analysis of urinary 4-pyridoxic acid (4-PA) by gas chromatography-mass spectrometry (GC-MS) for use in metabolic studies. Aliquots of urine were deproteinised and fractionated by isocratic reversed-phase high-performance liquid chromatography. The eluent fraction containing the 4-PA was collected, freeze-dried and silylated using N-methyl-N-(tert.-butyldimethylsilyl)trifluoroacetamide. Derivatisation produced the mono-tert.-butyldimethylsilyl derivative of 4-PA lactone. This derivative was readily amenable to GC-MS analysis in the electron ionisation (70 eV) mode, yielding a prominent fragment ion at m/z 222 ([M-57]+; base peak). A heavy isotope-labelled derivative of pyridoxine [dideuteriated pyridoxine; 3-hydroxy-4-(hydroxymethyl)-5-[hydroxymethyl-2H2]-2-methylpyridine] has been synthesised and is being employed to determine the kinetics of labelling of the body pools of vitamin B6. Kinetic measurements are based on the determination of the relative proportions of metabolically produced deuterium-labelled and non-labelled 4-PA in urine, obtained from stable isotope ratios determined by low-resolution selected ion monitoring using a bench-top quadrupole GC-MS system. PMID:1452608

Leyland, D M; Evershed, R P; Edwards, R H; Beynon, R J



One-step refolding and purification of recombinant human tumor necrosis factor-? (rhTNF-?) using ion-exchange chromatography.  


Protein refolding is a key step for the production of recombinant proteins, especially at large scales, and usually their yields are very low. Chromatographic-based protein refolding techniques have proven to be superior to conventional dilution refolding methods. High refolding yield can be achieved using these methods compared with dilution refolding of proteins. In this work, recombinant human tumor necrosis factor-? (rhTNF-?) from inclusion bodies expressed in Escherichia coli was renatured with simultaneous purification by ion exchange chromatography with a DEAE Sepharose FF column. Several chromatographic parameters influencing the refolding yield of the denatured/reduced rhTNF-?, such as the urea concentration, pH value and concentration ratio of glutathione/oxidized glutathione in the mobile phase, were investigated in detail. Under optimal conditions, rhTNF-? can be renatured and purified simultaneously within 30 min by one step. Specific bioactivity of 2.18?×?10(8) IU/mg, purity of 95.2% and mass recovery of 76.8% of refolded rhTNF-? were achieved. Compared with the usual dilution method, the ion exchange chromatography method developed here is simple and more effective for rhTNF-? refolding in terms of specific bioactivity and mass recovery. Copyright © 2014 John Wiley & Sons, Ltd. PMID:24941919

Wang, Yan; Ren, Wenxuan; Gao, Dong; Wang, Lili; Yang, Ying; Bai, Quan



Universal anion detection by replacement-ion chromatography with an atmospheric-pressure solution-cathode glow discharge photometric detector.  


A new method of detection for ion chromatography (IC) is described that couples replacement-ion chromatography (RIC) with an atmospheric-pressure solution-cathode glow discharge (SCGD). In the new instrument, a conventional suppressed IC arrangement is followed by a "replacement column" that consists of a cation-exchange micromembrane suppressor continuously regenerated with Li(2)SO(4). In this arrangement analytes are stoichiometrically converted to Li salts when they pass through the replacement column and are introduced into the SCGD, where they are detected indirectly by atomic emission spectrometry. Though found to be unsuitable for cation determinations, the SCGD-RIC instrument shows good repeatability (<3.5% peak area relative standard deviation), approximately 4 orders of linear dynamic range, universal calibration (similar molar sensitivity for anions of the same charge), and detection limits between 0.08 and 0.64 ?g·mL(-1) (25-?L injection) for several mono- and divalent anions. Deviations from the universal calibration were also observed and are critically evaluated. Optimal operating conditions are described, analytical figures of merit are presented, and background sources present in the system are characterized and explained. PMID:23157551

Schwartz, Andrew J; Wang, Zheng; Ray, Steven J; Hieftje, Gary M



Online spectrophotometric determination of Fe(II) and Fe(III) by flow injection combined with low pressure ion chromatography.  


A simple and new low pressure ion chromatography combined with flow injection spectrophotometric procedure for determining Fe(II) and Fe(III) was established. It is based on the selective adsorption of low pressure ion chromatography column to Fe(II) and Fe(III), the online reduction reaction of Fe(III) and the reaction of Fe(II) in sodium acetate with phenanthroline, resulting in an intense orange complex with a suitable absorption at 515nm. Various chemical (such as the concentration of colour reagent, eluant and reductive agent) and instrumental parameters (reaction coil length, reductive coil length and wavelength) were studied and were optimized. Under the optimum conditions calibration graph of Fe(II)/Fe(III) was linear in the Fe(II)/Fe(III) range of 0.040-1.0mg/L. The detection limit of Fe(III) and Fe(II) was respectively 3.09 and 1.55?g/L, the relative standard deviation (n=10) of Fe(II) and Fe(III) 1.89% and 1.90% for 0.5mg/L of Fe(II) and Fe(III) respectively. About 2.5 samples in 1h can be analyzed. The interfering effects of various chemical species were studied. The method was successfully applied in the determination of water samples. PMID:25523043

Chen, Shujuan; Li, Nan; Zhang, Xinshen; Yang, Dongjing; Jiang, Heimei



Multielement characterization of metal-humic substances complexation by size exclusion chromatography, asymmetrical flow field-flow fractionation, ultrafiltration and inductively coupled plasma-mass spectrometry detection: a comparative approach.  


The use of three different separation techniques, ultrafiltration (UF), high performance size exclusion chromatography (HPSEC) and asymmetrical flow field-flow fractionation (AsFlFFF), for the characterization of a compost leachate is described. The possible interaction of about 30 elements with different size fractions of humic substances (HS) has been investigated coupling these separation techniques with UV-vis absorption spectrophotometry and inductively coupled plasma-mass spectrometry (ICP-MS) as detection techniques. The organic matter is constituted by a polydisperse mixture of humic substances ranging from low molecular weights (around 1kDa) to significantly larger entities. Elements can be classified into three main groups with regard to their interaction with HS. The first group is constituted by primarily the monovalent alkaline metal ions and anionic species like B, W, Mo, As existing as oxyanions all being not significantly associated to HS. The second group consists of elements that are at least partly associated to a smaller HS size fraction (such as Ni, Cu, Cr and Co). A third group of mainly tri- and tetravalent metal ions like Al, Fe, the lanthanides, Sn and Th are rather associated to larger-sized HS fractions. The three separation techniques provide a fairly consistent size classification for most of the metal ions, even though slight disagreements were observed. The number-average molecular weight (Mn), the weight-average molecular weight (Mw) and the polydispersity (rho) parameters have been calculated both from AsFlFFF and HPSEC experiments and compared for HS and some metal-HS species. Differences in values can be partly explained by an overloading effect observed in the AsFlFFF experiments induced by electrostatic repulsion effects in the low ionic strength, high pH carrier solution. Size information obtained from ultrafiltration is not as resolved as for the other methods. Molecular weight cut-offs (MWCO) of the individual filter membranes refer to globular proteins and molecular weight information may therefore, deviate from that given by the other methods after calibration with polystyrene sulfonate (PSS) standards. PMID:16859692

Bolea, E; Gorriz, M P; Bouby, M; Laborda, F; Castillo, J R; Geckeis, H




EPA Science Inventory

The choice of gas chromatography (GC) detectors has expanded rapidly. The necessity for mass spectrometric (MS) characterization of GC effluents stems from the complexity of the matrices associated with environmental samples. There are currently several MS types being used in con...


Characterizing string-of-pearls colloidal silica by multidetector hydrodynamic chromatography and comparison to multidetector size-exclusion chromatography, off-line multiangle static light scattering, and transmission electron microscopy.  


The string-of-pearls-type morphology is ubiquitous, manifesting itself variously in proteins, vesicles, bacteria, synthetic polymers, and biopolymers. Characterizing the size and shape of analytes with such morphology, however, presents a challenge, due chiefly to the ease with which the "strings" can be broken during chromatographic analysis or to the paucity of information obtained from the benchmark microscopy and off-line light scattering methods. Here, we address this challenge with multidetector hydrodynamic chromatography (HDC), which has the ability to determine, simultaneously, the size, shape, and compactness and their distributions of string-of-pearls samples. We present the quadruple-detector HDC analysis of colloidal string-of-pearls silica, employing static multiangle and quasielastic light scattering, differential viscometry, and differential refractometry as detection methods. The multidetector approach shows a sample that is broadly polydisperse in both molar mass and size, with strings ranging from two to five particles, but which also contains a high concentration of single, unattached "pearls". Synergistic combination of the various size parameters obtained from the multiplicity of detectors employed shows that the strings with higher degrees of polymerization have a shape similar to the theory-predicted shape of a Gaussian random coil chain of nonoverlapping beads, while the strings with lower degrees of polymerization have a prolate ellipsoidal shape. The HDC technique is contrasted experimentally with multidetector size-exclusion chromatography, where, even under extremely gentle conditions, the strings still degraded during analysis. Such degradation is shown to be absent in HDC, as evidenced by the fact that the molar mass and radius of gyration obtained by HDC with multiangle static light scattering detection (HDC/MALS) compare quite favorably to those determined by off-line MALS analysis under otherwise identical conditions. The multidetector HDC results were also comparable to those obtained by transmission electron microscopy (TEM). Unlike off-line MALS or TEM, however, multidetector HDC is able to provide complete particle analysis based on the molar mass, size, shape, and compactness and their distributions for the entire sample population in less than 20 min. PMID:21428298

Brewer, Amandaa K; Striegel, André M



Liquid chromatography and ion trap mass spectrometry for simultaneous and multiclass analysis of antimicrobial residues in feed water.  


This work firstly reported the development of liquid chromatography coupled to an ion trap mass spectrometer (LC-MS ion trap) for the simultaneous determination of nitrofurans (e.g. nitrofurazone (NFZ), nitrofurantoin (NFT), furazolidone (FZD) and furaltadone (FTD)), nitroimidazoles (e.g. metronidazole (MNZ), ronidazole (RNZ) and dimetridazole (DMZ)) and chloramphenicol (CAP) in feed water. Isotope-labeled internal standards for the corresponding target analytes were employed to prevent matrix effects that might lead to signal suppression/enhancement. High performance liquid chromatography (HPLC) analysis was performed on a Prodigy ODS-3 column, 2.0mm×150mm, 5?m with a guard cartridge at a flow rate of 0.2mL/min, column oven temperature of 40°C, and an injection volume of 10?L. Solid phase extraction (SPE) procedures, factors affecting HPLC separation (e.g. buffer pH and concentrations) and mass spectrometry (MS) parameters were optimized. After an off-line SPE by the OASIS HLB cartridges (with an enrichment factor of 400), the eight antimicrobial agents were separated in 18min using a gradient elution of acetonitrile in acidified water (pH 5.0). MS detection was by an ion trap MS coupled with electrospray ionization (ESI) in tandem mass spectrometry mode (MS/MS) using the nebulizer gas at 35psi, drying gas at 9L/min and drying temperature of 325°C. Method linearity was good (r(2)=0.979-0.999) with acceptable precision (% RSDs=3.4-26.6%) and accuracy (%recovery=88.4-110.1%). Very low limits of detection (LOD) and quantitation (LOQ) were achieved in ranges of 0.002-0.06?g/L and 0.005-0.25?g/L, respectively. The established method is successfully employed by the Department of Livestock Development of Thailand for the monitoring of the drug residues in feed waterbecause of its convenience, reliability and high sensitivity. PMID:24321758

Ardsoongnearn, Chusak; Boonbanlu, Ongart; Kittijaruwattana, Sunan; Suntornsuk, Leena




Microsoft Academic Search

SUMMARY The equilibrium distribution coefficients (p-value) of D-arabinose between the water in sulfonate ion-exchange resin and the external aqueous solution vary with the nature of the five alkali metal counterions studied. The strongest exclusion (lowest p -value) is found in the Li+ resin and the least exclusion (highest p -value) in the Cs+ resin. The p-value decreases with the increasing



Speciation of heavy metals in environmental water by ion chromatography coupled to ICP-MS  

Microsoft Academic Search

Biogenic (e.g. phytochelatins, porphyrins, DOM) as well as anthropogenic (e.g. NTA, EDTA, phosphonates) chelators affect the mobility and cycling of heavy metals in environmental waters. Since such chelators can form strongly bound anionic heavy metal complexes that are stable and highly mobile, anion-exchange chromatography coupled to ICP-MS was investigated. A narrow bore HPLC system was connected to a micro concentric

Adrian A. Ammann




EPA Science Inventory

The ion-exchange separation theory of Mayer and Tompkins is extended to the case of multiple eluent ions. The selectivity coefficients for carbonate and hydroxide, each relative to bicarbonate, were estimated from retention data for bromide and sulfate. These results were used to...


How does roasting affect the antioxidants of a coffee brew? Exploring the antioxidant capacity of coffee via on-line antioxidant assays coupled with size exclusion chromatography.  


During coffee roasting major changes occur in coffee bean composition. Among others dark coloured melanoidins are formed, which are high molecular weight Maillard reaction products. A new approach is presented here to monitor the influence of roasting conditions on the antioxidant capacity of melanoidins and chlorogenic acids (CGAs) in a coffee brew. Validated Folin-Ciocalteu (FC) and ABTS assays were used as on-line antioxidant assays coupled (post-column) with high performance size-exclusion chromatography (HPSEC). HPSEC enabled the separation of melanoidins from CGAs and the determination of the antioxidant capacity of each fraction, within a total elution time of 25 min. Besides the on-line assay measurements, both assays were also applied off-line with flow injection analysis (FIA). The maximum antioxidant capacity was determined to be at a light-to-medium roast degree, measured with both ABTS-FIA and FC-FIA assays as well as on-line ABTS assay. With FC on-line assay the maximum was found to be at a very light roast degree. Based on the peak areas obtained with the new coupled technique the roasting effects on the variability of melanoidin and CGA contents in coffee brews were studied. The majority of melanoidins are already formed in the early stage of the roasting process and the relative contribution of melanoidins to the total antioxidant capacity increases towards darker roasts, mainly because CGAs degrade during roasting. A new parameter, the ratio of melanoidin to CGA peak area, was introduced as a possible predictor of the roast degree. PMID:23592006

Smrke, Samo; Opitz, Sebastian E W; Vovk, Irena; Yeretzian, Chahan



Characterization of the molar masses of hemicelluloses from wood and pulps employing size exclusion chromatography and matrix-assisted laser desorption ionization time-of-flight mass spectrometry.  


The molar mass parameters for arabino-4-O-methylglucuronoxylans, arabinohexenuronoxylans, 4-O-methylglucuronoxylans, hexenuronoxylans, and galactoglucomannans extracted from wood and pulps have been determined. To characterize different types of hemicelluloses, delignified wood (spruce, pine, larch, aspen, and birch) and chemical pulps (unbleached and totally chlorine-free bleached) were extracted with dimethyl sulfoxide (DMSO) or alkaline aqueous solutions. Size exclusion chromatography (SEC) with off-line matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-MS) were employed to characterize the molar masses. The hemicellulose extracts were separated by SEC into fractions each containing components with a narrow range of molar masses and the average molar mass of each fraction subsequently determined by MALDI-MS. The molar mass parameters for the hemicelluloses were then calculated on the basis of the SEC distribution curves and MALDI-MS spectra. As expected, in most cases the hemicelluloses extracted from wood (holocellulose) exhibited higher molar masses than did the corresponding hemicelluloses from chemical pulps. The molar mass parameters for hemicelluloses isolated from pulps derived from cooking samples of the same batch of softwood chips decreased in the following order: ASAM pulp > MSSAQ pulp > kraft pulp. The lowest molar masses were demonstrated by the glucuronoxylans extracted from pulps obtained by cooking with acidic sulfite. The xylans from bleached kraft pulp were characterized by molar masses that were only slightly lower than those of the corresponding xylans from unbleached pulp. The xylans extracted into DMSO exhibited somewhat lower molar masses than did the corresponding xylans extracted into alkaline aqueous solutions. In all cases the range of molar masses demonstrated by the hemicelluloses investigated was found to be rather narrow, i.e., the polydispersity index Mw/Mn was found to be approximately 1.1-1.4. PMID:11710047

Jacobs, A; Dahlman, O



Generic detection of basic taxoids in wood of European Yew (Taxus baccata) by liquid chromatography-ion trap mass spectrometry.  


The occurrence of the cardiotoxin taxine (comprising taxine B and several other basic taxoids) in leaves of Taxus baccata L. (European yew) is well known and has led to public concerns about the safety of eating or drinking from utensils crafted from the wood of this poisonous species. The occurrence of basic taxoids in the heartwood of T. baccata had not been examined in detail, although the bark is known to contain 2'?-deacetoxyaustrospicatine. Initial examination of heartwood extracts for 2'?-deacetoxyaustrospicatine by liquid chromatography-mass spectrometry (LC-MS) revealed the presence of this basic taxoid at about 0.0007% dry weight, using a standard isolated from bark. Analyses for taxine B, however, proved negative at the extract concentration analysed. Observing other basic taxoids within the heartwood extracts was facilitated by developing generic LC-MS methods that utilised a fragment arising from the N-containing acyl group of basic taxoids as a reporter ion. Of the various MS strategies available on a hybrid ion trap-orbitrap instrument that allowed observation of this reporter ion, combining all-ion collisions with high resolution ion filtering by the orbitrap was most effective, both in terms of the number of basic taxoids detected and sensitivity. Numerous basic taxoids, in addition to 2'?-deacetoxyaustrospicatine, were revealed by this method in heartwood extracts of T. baccata. Red wine readily extracted the basic taxoids from heartwood while coffee extracted them less efficiently. Contamination with basic taxoids could also be detected in soft cheese that had been spread onto wood. The generic LC-MS method for detecting basic taxoids complements specific methods for detecting taxine B when investigating yew poisoning cases in which the analysis of complex extracts may be required or taxine B has not been detected. PMID:23314400

Kite, Geoffrey C; Rowe, Emily R; Veitch, Nigel C; Turner, Jill E; Dauncey, Elizabeth A



Determination of fluorine, chlorine and bromine in household products by means of oxygen bomb combustion and ion chromatography.  


A method for routine determination of fluorine, chlorine and bromine in household products was developed and validated. In this work, halogen analyses were made based on oxygen bomb combustion followed by ion chromatography (IC). The chromatographic analysis was performed by an IonPac AS19 hydroxide-selective anion-exchange column, a reagent free ion chromatograph eluent generator and an anion self-regenerating suppressor in 10 min. The response was linear (r ? 0.9995) in the entire investigated domain. The limit of detection for the halogens was in the range of 2 to 9 × 10(-3) mg/L and the limit of quantification was lower than 8 mg/Kg with 20 µL of injection volume. The certified reference material of ERM-EC 681k was pretreated using an oxygen bomb combustion procedure to demonstrate the precision of the proposed method. The quantitative analysis results obtained by IC for the target elements were 797 ± 9 mg/Kg chlorine and 786 ± 25 mg/Kg bromine, which were in good agreement with the certified values of 800 ± 4 mg/Kg chlorine, 770 ± 5 mg/Kg bromine for ERM-EC 681k, respectively. This validated method was successfully applied for the analysis of fluorine, chlorine and bromine in household product samples, and the variation of halogen contained among the tested samples was remarkable. PMID:22752184

Zhang, Shuai; Zhao, Tianbo; Wang, Jia; Qu, Xiaoling; Chen, Wei; Han, Yin



Determination of vinyl chloride in soil-gas samples by gas chromatography coupled with ion mobility spectrometry  

SciTech Connect

The advantages of ion mobility spectrometry (IMS), such as ambient pressure operation, simplicity of design, high sensitivity, speed of response, spectral output, and ability to respond to a large number of compounds, make IMS technology attractive for a variety of field monitoring applications. A quantitative, multicompound field detector based on IMS technology has not been successful, however, because of technology limitations. For multicomponent samples, multiple ion-molecule reactions result in confusing spectral information. Varying humidity can result in ion-water clusters that cause errors in both the identification and quantification of the target analyte. These limitations could be overcome through the addition of gas chromatography (GC) prior to high-temperature IMS. When coupling GC to IMS, the following need to be considered: (a) currently available IMS cells have a relatively large internal volume (> 20 mL), reducing the chromatographic resolution; (b) smaller IMS cells are not capable of operating above 100 C; (c) the radioactive ionization cell limits the dynamic range of the device to 2 to 3 orders of magnitude; and (d) most standard miniature cells have a membrane inlet that does not allow direct interface with a GC column. The authors have overcome some of these difficulties by interfacing GC with Fourier Transform IMS (FTIMS). They will present data on determining vinyl chloride in soil-gas samples by GC-FTIMS.

Jones, T.L. [Environmental Protection Agency, Las Vegas, NV (United States); Hill, H.; Simpson, G.; Klasmeier, M. [Washington State Univ., Pullman, WA (United States); Lopez-Avila, V. [Midwest Research Inst., Mountain View, CA (United States); Radolovich, G. [Midwest Research Inst., Kansas City, MO (United States)



Sensitive bioassay for the simultaneous determination of pseudoephedrine and cetirizine in human plasma by liquid-chromatography-ion trap spectrometry.  


A liquid chromatography-ion trap mass spectrometry coupled with electrospray ionization (HPLC-ESI-ion trap mass spectrometry) method for simultaneous determination of cetirizine and pseudoephedrine in human plasma is presented. Chromatographic separation was performed on a Hypurity C18 column (Thermo Hypersil-Keystone 2.1 mm x 150 mm, 5 microm, USA), The mobile phase was composed of 65% methanol and 35% water (contained 0.1% formic acid, 10 mM ammonium formate), which was run with a flow-rate of 0.2 ml/min at 40 degrees C. Quantitation was achieved by monitoring the product ions at m/z 166-->m/z 148 (pseudoephedrine), m/z 389.9-->m/z 201.1 (cetirizine), m/z 264-->m/z 246 (tramadol, IS). The calibration curve of pseudoephedrine and cetirizine was established with standard solutions. The limit of detection for pseudoephedrine and cetirizine each was 5 ng/ml. This simplified analytical method is sensitive, specific and accurate enough for simultaneous determination of pseudoephedrine and cetirizine in human plasma and is successfully applied to the pharmacokinetic study of pseudoephedrine and cetirizine. PMID:16713697

Tan, Zhi-Rong; Ouyang, Dong-Sheng; Zhou, Gan; Wang, Lian-Sheng; Li, Zhi; Wang, Dan; Zhou, Hong-Hao



Metabolomic analysis via reversed-phase ion-pairing liquid chromatography coupled to a stand alone orbitrap mass spectrometer  

PubMed Central

We present a liquid chromatography-mass spectrometry (LC-MS) method that capitalizes on the mass-resolving power of the orbitrap to enable sensitive and specific measurement of known and unanticipated metabolites in parallel, with a focus on water soluble species involved in core metabolism. The reversed phase LC method, with a cycle time 25 min, involves a water-methanol gradient on a C18 column with tributylamine as the ion pairing agent. The MS portion involves full scans from 85 – 800 m/z at 1 Hz and 100,000 resolution in negative ion mode on a stand alone orbitrap (“Exactive”). The median limit of detection, across 80 metabolite standards, was 5 ng/mL with linear range typically ? 100-fold. For both standards and a cellular extract from Saccharomyces cerevisiae (Baker’s yeast), the median inter-run relative standard deviation in peak intensity was 8%. In yeast exact, we detected 137 known compounds, whose 13C-labeling patterns could also be tracked to probe metabolic flux. In yeast engineered to lack a gene of unknown function (YKL215C), we observed accumulation of an ion of m/z 128.0351, which we subsequently confirmed to be oxoproline, resulting in annotation of YKL215C as an oxoprolinase. These examples demonstrate the suitability of the present method for quantitative metabolomics, fluxomics, and discovery metabolite profiling. PMID:20349993

Lu, Wenyun; Clasquin, Michelle F.; Melamud, Eugene; Amador-Noguez, Daniel; Caudy, Amy A.; Rabinowitz, Joshua D.



Nano TiO 2-based preconcentration for the speciation analysis of inorganic selenium by using ion chromatography with conductivity detection  

Microsoft Academic Search

A simple, sensitive, and cost-effective analytical method was developed for the speciation analysis of inorganic selenium by combining a nano-TiO2 preconcentration with an ion chromatography-conductivity detection (IC-CD) system. The experimental conditions for the simultaneous adsorption and desorption of Se(IV) and Se(VI) were carefully investigated. Under the established optimum condition, the Se(IV) and Se(VI) ions could been simultaneously adsorbed onto the

Shuxia Xu; Maoli Zheng; Xinfeng Zhang; Junlong Zhang; Yong-Ill Lee


Simultaneous determination of fatty, dicarboxylic and amino acids based on derivatization with isobutyl chloroformate followed by gas chromatography—positive ion chemical ionization mass spectrometry  

Microsoft Academic Search

Gas chromatography–mass spectrometry (GC–MS) with positive ion chemical ionization (PICI) using isobutane as reagent gas was applied for analysis of isobutoxycarbonyl\\/isobutyl derivatives of 13 fatty, 6 dicarboxylic and 13 amino acids in a single run. For all investigated compounds (except several amino acids) the quasimolecular ions [MH]+ were registered. Asparagine underwent fragmentation via decarboxylation followed by elimination of OC4H9 ([M?117]+),

Tim G. Sobolevsky; Alexander I. Revelsky; Igor A. Revelsky; Barbara Miller; Vincent Oriedo



A solid phase extraction-ion chromatography with conductivity detection procedure for determining cationic surfactants in surface water samples.  


A new analytical procedure for the simultaneous determination of individual cationic surfactants (alkyl benzyl dimethyl ammonium chlorides) in surface water samples has been developed. We describe this methodology for the first time: it involves the application of solid phase extraction (SPE-for sample preparation) coupled with ion chromatography-conductivity detection (IC-CD-for the final determination). Mean recoveries of analytes between 79% and 93%, and overall method quantification limits in the range from 0.0018 to 0.038 ?g/mL for surface water and CRM samples were achieved. The methodology was applied to the determination of individual alkyl benzyl quaternary ammonium compounds in environmental samples (reservoir water) and enables their presence in such types of waters to be confirmed. In addition, it is a simpler, less time-consuming, labour-intensive, avoiding use of toxic chloroform and significantly less expensive methodology than previously described approaches (liquid-liquid extraction coupled with liquid chromatography-mass spectrometry). PMID:24148395

Olkowska, Ewa; Polkowska, ?aneta; Namie?nik, Jacek



Identification and quantitation of auxins in plants by liquid chromatography/electrospray ionization ion trap mass spectrometry.  


Auxin is an important phylohormone, which regulates specific physiological responses such as division, elongation and differentiation of cells. A new method using liquid chromatography/electrospray ionization ion trap mass spectrometry (LC/ESI-ITMS) has been developed for identification and quantitation of four auxins. Under the optimum conditions, four auxins (indole-3-acetic acid, indole-3-propionic acid, indole-3-butyric acid and 1-naphthylacetic acid) were completely separated and quantitated within 7 min with a minimum detection limit of 8.0 ng mL(-1) with relative standard deviations lower than 5.0%. This method also has been applied to analysis of auxins in Chinese cabbage where, even with a complicated serious background perturbation due to the natural biological matrix, the mean recoveries ranged from 77.5% to 99.8%. Finally, we discuss the MS-relevant properties of the identified auxins in detail. PMID:18655000

Lu, Qiaomei; Zhang, Lan; Chen, Tianwen; Lu, Minghua; Ping, Tong; Chen, Guonan



Hand-portable gas chromatography-ion mobility spectrometer for the determination of the freshness of fish  

NASA Technical Reports Server (NTRS)

A hand-held, portable gas chromatography-ion mobility spectrometer (GC-IMS) device was used to detect the presence of volatile amine compounds in the headspace of decomposing fish. The Food and Drug Administration (FDA) largely relies on olfactory discrimination with respect to fresh and spoiled, frozen and unfrozen fish. The fish are delivered at ship docks on pallets, and each pallet of fish can range from 30-40 thousand dollars in value. Fresh fish were placed in a teflon bag and the direct headspace was interrogated. In the first three days, only low molecular weight volatile amines were detected. On the fourth day, a number of spectral signatures were observed which indicated the presence of 1,5-diaminopentane, cadaverine. Analyses typically took from 0.5-1 minute.

Snyder, A. Peter; Harden, Charles S.; Davis, Dennis M.; Shoff, Donald B.; Maswadeh, Waleed M.




EPA Science Inventory

The objective of this research was to evaluate, in the laboratory, the potential of gas chromatography/ion mobility spectrometry (GC/IMS) for monitoring vinyl chloride and other organic compounds in air samples in the field. It was determined that GC/IMS has the potential to dire...



EPA Science Inventory

Musk xylene (MX) is widely used as a fragrance ingredient in commercial toiletries. Identification and quantification of a bound 4-amino-MX (AMX) metabolite was carried out by gas chromatography-mass spectrometry (GC/MS), with selected ion monitoring (SIM). Detection of AMX occur...


Profiling the indole alkaloids in yohimbe bark with ultra-performance liquid chromatography coupled with ion mobility quadrupole time-of-flight mass spectrometry  

Technology Transfer Automated Retrieval System (TEKTRAN)

An ultra-high performance liquid chromatography-ion mobility- quadrupole time-of-flight mass spectrometry (UHPLC-IM-QTOF-MS) method was developed for profiling the indole alkaloids in yohimbe bark. Many indole alkaloids with the yohimbine core structure, plus methylated, oxidized, and reduced speci...


A Computer-Based Undergraduate Exercise Using Internet-Accessible Simulation Software for the Study of Retention Behavior and Optimization of Separation Conditions in Ion Chromatography  

ERIC Educational Resources Information Center

The ability to scan retention data over a wide range of eluent composition opens up the possibility of a computerized selection of the optimal separation conditions. The major characteristics of retention behavior, peak-shape effects and pH effects evident in ion chromatography (IC) using common stationary phases and eluents are illustrated.

Haddad, Paul R.; Shaw, Matthew J.; Madden, John E.; Dicinoski, Greg W.



Comparison of liquid chromatography using triple quadrupole and quadrupole ion trap mass analyzers to determine pesticide residues in oranges.  


Liquid chromatography-triple quadrupole/mass spectrometry (LC-TQ/MS) and liquid chromatography-quadrupole ion trap/mass spectrometry (LC-QIT/MS) for determining bupirimate, hexaflumuron, tebufenpyrad, buprofezin, pyriproxyfen, and fluvalinate in fruits have been compared. The differences in the mass spectra obtained by triple and ion trap quadrupoles are discussed, showing how both of them provide interesting features. The evaluation of the two instruments was carried out by ethyl acetate extraction of oranges spiked with the studied pesticides at LOQ and 10 times the LOQ. Results obtained by LC-TQ/MS correlated well with those obtained by LC-QIT/MS. Recoveries were 70-94% by LC-TQ/MS and 72-92% by LC-QIT/MS with the R.S.D. from five replicate analysis 4-14% and 8-18%, respectively. Matrix effects were tested for both techniques by standard addition to blank extracts. Although the matrix effects are not originated in mass analyzer but in the LC/MS interface, they were, generally, more marked by LC-QIT-MS than by LC-TQ/MS. The limits of quantification (LOQs) were 0.005-0.2 mg kg(-1) by both equipments--appropriate values for determining these pesticides in orange from the regulatory point of view. The results indicate that the TQ provides higher precision, better linearity, it is more robust, and when the purpose of the analysis is quantitative determination, preferable over the QIT. However, the application of both mass spectrometers to analyze orange samples conventionally treated showed that any can be used for qualitative and quantitative purposes. PMID:15844516

Soler, Carla; Mañes, Jordi; Picó, Yolanda



Liquid chromatography quadrupole linear ion trap mass spectrometry for multiclass screening and identification of lipophilic marine biotoxins in bivalve mollusks.  


A liquid chromatography quadrupole linear ion trap mass spectrometry method with fast polarity switching and a scheduled multiple reaction monitoring algorithm mode was developed for multiclass screening and identification of lipophilic marine biotoxins in bivalve molluscs. A major advantage of the method is that it can detect members of all six groups of lipophilic marine biotoxins [okadaic acid (OA), yessotoxins (YTX), azaspiracids (AZA), pectenotoxins (PTX), cyclic imines (CI), and brevetoxins (PbTx)], thereby allowing quantification and high confidence identification from a single liquid chromatography tandem mass spectrometry (LC-MS/MS) injection. An enhanced product ion (EPI) library was constructed after triggered collection of data via information-dependent acquisition (IDA) of EPI spectra from standard samples. A separation method for identifying 17 target toxins in a single analysis within 12min was developed and tested. Different solid phase extraction sorbents, the matrix effect (for oyster, scallop, and mussel samples), and stability of the standards also were evaluated. Matrix-matched calibration was used for quantification of the toxins. The limits of detection were 0.12-13.6?g/kg, and the limits of quantification were 0.39-45.4?g/kg. The method was used to analyze 120 shellfish samples collected from farming areas along the coast of China, and 7% of the samples were found to be contaminated with toxins. The library search identified PbTx-3, YTX, OA, PTX2, AZA1, AZA2, and desmethylspirolide C (SPX1). Overall, the method exhibited excellent sensitivity and reproducibility, and it will have broad applications in the monitoring of lipophilic marine biotoxins. PMID:25086754

Wu, Haiyan; Guo, Mengmeng; Tan, Zhijun; Cheng, Haiyan; Li, Zhaoxin; Zhai, Yuxiu



Simultaneous determination of triprolidine and pseudoephedrine in human plasma by liquid chromatography-ion trap mass spectrometry.  


A highly efficient, selective and specific method for simultaneous quantitation of triprolidine and pseudoephedrine in human plasma by liquid chromatography-ion trap-tandem mass spectrometry coupled with electro spray ionization (LC-ESI-ion trap-tandem MS) has been validated and successfully applied to a clinical pharmacokinetic study. Both targeted compounds together with the internal standard (gabapentin) were extracted from the plasma by direct protein precipitation. Chromatographic separation was achieved on a C(18) ACE((R)) column (50.0mmx2.1mm, 5mum, Advance Chromatography Technologies, Aberdeen, UK), using an isocratic mobile phase, consisting of water, methanol and formic acid (55:45:0.5, v/v/v), at a flow-rate of 0.3mL/min. The transition monitored (positive mode) was m/z 279.1-->m/z 208.1 for triprolidine, m/z 165.9-->m/z 148.0 for pseudoephedrine and m/z 172.0-->m/z 154.0 for gabapentin (IS). This method had a chromatographic run time of 5.0min and a linear calibration curves ranged from 0.2 to 20.0ng/mL for triprolidine and 5.0-500.0ng/mL for pseudoephedrine. The within- and between-batch accuracy and precision (expressed as coefficient of variation, %C.V.) evaluated at four quality control levels were within 94.3-106.3% and 1.0-9.6% respectively. The mean recoveries of triprolidine, pseudoephedrine and gabapentin were 93.6, 76.3 and 82.0% respectively. Stability of triprolidine and pseudoephedrine was assessed under different storage conditions. The validated method was successfully employed for the bioequivalence study of triprolidine and pseudoephedrine formulation in twenty six volunteers under fasting conditions. PMID:19926351

Shakya, Ashok K; Arafat, Tawfiq A; Abuawwad, Ahmad N; Melhim, Munther; Al-Ghani, Jafar; Yacoub, Mahmoud J



Characterization of diacylglycerol isomers in edible oils using gas chromatography-ion trap electron ionization mass spectrometry.  


Verifying the authenticity of edible oils is of international concern. A new quality control standard for olive oil has been proposed that relates the ratio of 1,2-diacylglycerol (DAG) to 1,3-DAG to sensory aspects of olive oil. DAGs and their isomers are difficult to quantitate and characterize by Flame Ionization Gas Chromatography (GC-FID) due to the lack of suitable standards. Mass detectors offer the advantage of providing structural detail to the eluding DAG(s), thus removing ambiguity to the identification of both resolved and unresolved DAGs in GC chromatograms. In this study, a GC Electron Ionization Mass Spectrometry (GC-EI-MS) method was developed to determine the fatty acid composition and molecular structure of trimethylsilyl (TMS) derivatized DAGs present in edible oils. Twenty-two species of DAG isomers were identified in refined coconut oil and unrefined olive oil utilizing signature fragment ions, [M-15](+), [M-89](+), [M-RCO2](+), [RCO2+58](+) and [M-RCO2CH2](+). The [M-RCO2CH2](+) ion is considered the key diagnostic ion to distinguish between DAG positional isomers. MS/MS spectra of [M-RCO2](+) and [M-15](+) ions obtained from commercial standards containing both 1,2- and 1,3-DAG isomers were used as a model system to confirm the identification of DAG isomers in natural products. Furthermore, a number of reaction mechanisms are proposed to explain the formation of the most abundant mass fragments of DAGs and their isomers. PMID:23880469

Zhu, Hanjiang; Clegg, Michael S; Shoemaker, Charles F; Wang, Selina C



Hydrodynamic Chromatography  

NASA Astrophysics Data System (ADS)

Hydrodynamic chromatography (HDC) has experienced a resurgence in recent years for particle and polymer characterization, principally because of its coupling to a multiplicity of physical detection methods. When coupled to light scattering (both multiangle static and quasi-elastic), viscometric, and refractometric detectors, HDC can determine the molar mass, size, shape, and structure of colloidal analytes continuously and as a function of one another, all in a single analysis. In so doing, it exposes the analytes to less shear force (and, hence, less potential for flow-induced degradation) than in, for instance, size-exclusion chromatography. In this review, we discuss the fundamental chromatographic underpinnings of this technique in terms of retention, band broadening, and resolution, and we describe the power of multidetector HDC with examples from the recent literature.

Striegel, André M.; Brewer, Amandaa K.



Preparation of purified erythropoietin by high performance liquid chromatography.  


Highly purified erythropoietin was not available in quantities needed to carry out planned investigations and, therefore, the use of high performance liquid chromatography was explored. This technique permits the separation of proteins with high efficiency and resolution. Three types of chromatography were used. Size exclusion or gel permeation, reversed phase, and ion exchange columns were utilized with different solvent systems. The chromatographic fractions were assayed either by an exhypoxic polycythemic mouse assay or by the fetal liver cell assay. In addition, selected fractions were tested for their capability to stimulate CFU-E and BFU-E colony formation in methyl cellulose. The results of the techniques of size exclusion and ion exchange chromatography were found to be rapid and reproducible. Although reversed phase chromatography gave excellent resolution, the results were somewhat variable. Using different chromatographic combinations, erythropoietin with a specific bioactivity in the range of 50,000 u/mg protein was isolated. Although the erythropoietin gene has now been cloned and the hormone purified by utilizing monoclonal antibodies, high performance liquid chromatography may be useful in the removal of unwanted contaminants, as a tool in chemical characterization, and as a possible method for the hormone's identification and measurement in clinical laboratories. PMID:6543653

Lange, R D; Andrews, R B; Trent, D J; Reyniers, J P; Draganac, P S; Farkas, W R



Identification and separation of saxitoxins using hydrophilic interaction liquid chromatography coupled to traveling wave ion mobility-mass spectrometry.  


The aim of this work was to develop a reliable and efficient analytical method to characterise and differentiate saxitoxin analogues (STX), including sulphated (gonyautoxins, GTX) and non-sulphated analogues. For this purpose, hydrophilic interaction liquid chromatography (HILIC) was used to separate sulphated analogues. We also resorted to ion mobility spectrometry to differentiate the STX analogues because this technique adds a new dimension of separation based on ion gas phase conformation. Positive and negative ionisation modes were used for gonyautoxins while positive ionisation mode was used for non-sulphated analogues. Subsequently, the coupling of these three complementary techniques, HILIC-IM-MS, permitted the separation and identification of STX analogues; isomer differentiation was achieved in HILIC dimension while non-sulphated analogues were separated in the IM-MS dimension. Additional structural characteristics concerning the conformation of STXs could be obtained using IM-MS measurements. Thus, the collision cross sections (CCS) of STXs are reported for the first time in the positive ionisation mode. These experimental CCSs correlated well with the calculated CCS values using the trajectory method. Copyright © 2015 John Wiley & Sons, Ltd. PMID:25601690

Poyer, Salomé; Loutelier-Bourhis, Corinne; Coadou, Gaël; Mondeguer, Florence; Enche, Julien; Bossée, Anne; Hess, Philipp; Afonso, Carlos



The determination of the Fe sup 2+ /Fe sup 3+ ratio in simulated nuclear waste glass by ion chromatography  

SciTech Connect

Liquid high-level nuclear waste will be immobilized at the Savannah River Site (SRS) by vitrification in borosilicate glass in the Defense Waste Processing Facility (DWPF). In this facility, control of the oxidation/reduction (redox) equilibrium in the glass melter is critical for processing of the nuclear waste. Therefore, the development of a rapid and reliable analytical method for the determination of the redox equilibrium is of considerable interest. Redox has been determined by measuring the ratio of ferrous to ferric ions in the glass melt. Two analytical techniques for glass redox measurement have been investigated for the DWPF: Mossbauer Spectroscopy which may be subject to interferences from the radiation in actual waste, and a rapid and simple chemical dissolution/spectrophotometric technique. Comparisons of these techniques have been made at several laboratories including Clemson University. In the study attached, the determination of the redox ratio by Ion Chromatography (IC) was investigated as a potential new technology. Clemson University performed IC analyses on the same glasses as previously examined by wet chemical and Mossbauer techniques. Results from all three techniques were highly correlated and IC was reported to be a promising new technology for redox measurement. 19 refs., 19 figs., 5 tabs.

Jantzen, C.M.



Analysis of water from the Space Shuttle and Mir Space Station by ion chromatography and capillary electrophoresis  

NASA Technical Reports Server (NTRS)

Drinking water and condensate samples collected from the US Space Shuttle and the Russian Mir Space Station are analyzed routinely at the NASA-Johnson Space Center as part of an ongoing effort to verify water quality and monitor the environment of the spacecraft. Water quality monitoring is particularly important for the Mir water supply because approximately half of the water consumed is recovered from humidity condensate. Drinking water on Shuttle is derived from the fuel cells. Because there is little equipment on board the spacecraft for monitoring the water quality, samples collected by the crew are transported to Earth on Shuttle or Soyuz vehicles, and analyzed exhaustively. As part of the test battery, anions and cations are measured by ion chromatography, and carboxylates and amines by capillary electrophoresis. Analytical data from Shuttle water samples collected before and after several missions, and Mir condensate and potable recovered water samples representing several recent missions are presented and discussed. Results show that Shuttle water is of distilled quality, and Mir recovered water contains various levels of minerals imparted during the recovery processes as designed. Organic ions are rarely detected in potable water samples, but were present in humidity condensate samples.

Orta, D.; Mudgett, P. D.; Ding, L.; Drybread, M.; Schultz, J. R.; Sauer, R. L.



Determination of four tobacco-specific nitrosamines in mainstream cigarette smoke by gas chromatography/ion trap mass spectrometry.  


A specific and sensitive method was developed and validated to quantitatively analyze four tobacco-specific nitrosamines (TSNAs) in the particulate phase of mainstream cigarette smoke. Cigarette smoke particulate matter was collected according to ISO 4387. The particulate matter was extracted with acetic ether, cleaned up with a Supelclean ENVI-Carb silod-phase extraction (SPE) cartridge, concentrated under the protection of nitrogen and analyzed by gas chromatography (GC)/ion trap mass spectrometry (MS) with a very-low-flow-loss column (VF-17 MS) in MS(n) mode using N-nitrosopentyl-3-picolylamine (NNPA) as an internal standard. TSNAs were identified by chromatographic retention time, matching the spectra of the standards and the samples and the consistency of the ratio of the abundance of the ions detected in the standards and the samples. Limits of detection of each TSNA varied from 0.01 to 0.06 ng/cig. A linear calibration range for this method is large enough to measure TSNA levels in cigarette smoke. The recovery of each TSNA was from 91.5 to 102.7%. The method achieved excellent reproducibility (RSD: 1.8-4.8% for intra-assay, 3.4-7.1% for inter-assay). It also shows no evidence of artifact formation. This method is very suitable for the routine detection of TSNAs in mainstream cigarette smoke. PMID:18008390

Zhou, Jun; Bai, Ruoshi; Zhu, Yongfa



Ion chromatography-mass spectrometry: a review of recent technologies and applications in forensic and environmental explosives analysis.  


The development and application of ion chromatography (IC) coupled to mass spectrometry (MS) is discussed herein for the quantitative determination of low-order explosives-related ionic species in environmental and forensic sample types. Issues relating to environmental explosives contamination and the need for more confirmatory IC-MS based applications in forensic science are examined. In particular, the compatibility of a range of IC separation modes with MS detection is summarised along with the analytical challenges that have been overcome to facilitate determinations at the ng-?g L(-1) level. Observed trends in coupling IC to inductively coupled plasma and electrospray ionisation mass spectrometry form a particular focus. This review also includes a discussion of the relative performance of reported IC-MS methods in comparison to orthogonal ion separation-based, spectrometric and spectroscopic approaches to confirmatory detection of low-order explosives. Finally, some promising areas for future research are highlighted and discussed with respect to potential IC-MS applications. PMID:24331039

Barron, Leon; Gilchrist, Elizabeth



Development Of Ion Chromatography Methods To Support Testing Of The Glycolic Acid Reductant Flowsheet In The Defense Waste Processing Facility  

SciTech Connect

Ion Chromatography (IC) is the principal analytical method used to support studies of Sludge Reciept and Adjustment Tank (SRAT) chemistry at DWPF. A series of prior analytical ''Round Robin'' (RR) studies included both supernate and sludge samples from SRAT simulant, previously reported as memos, are tabulated in this report.2,3 From these studies it was determined to standardize IC column size to 4 mm diameter, eliminating the capillary column from use. As a follow on test, the DWPF laboratory, the PSAL laboratory, and the AD laboratory participated in the current analytical RR to determine a suite of anions in SRAT simulant by IC, results also are tabulated in this report. The particular goal was to confirm the laboratories ability to measure and quantitate glycolate ion. The target was + or - 20% inter-lab agreement of the analyte averages for the RR. Each of the three laboratories analyzed a batch of 12 samples. For each laboratory, the percent relative standard deviation (%RSD) of the averages on nitrate, glycolate, and oxalate, was 10% or less. The three laboratories all met the goal of 20% relative agreement for nitrate and glycolate. For oxalate, the PSAL laboratory reported an average value that was 20% higher than the average values reported by the DWPF laboratory and the AD laboratory. Because of this wider window of agreement, it was concluded to continue the practice of an additional acid digestion for total oxalate measurement. It should also be noted that large amounts of glycolate in the SRAT samples will have an impact on detection limits of near eluting peaks, namely Fluoride and Formate. A suite of scoping experiments are presented in the report to identify and isolate other potential interlaboratory disceprancies. Specific ion chromatography inter-laboratory method conditions and differences are tabulated. Most differences were minor but there are some temperature control equipment differences that are significant leading to a recommendation of a heated jacket for analytical columns that are remoted for use in radiohoods. A suggested method improvement would be to implement column temperture control at a temperature slightly above ambient to avoid peak shifting due to temperature fluctuations. Temperature control in this manner would improve short and longer term peak retention time stability. An unknown peak was observed during the analysis of glycolic acid and SRAT simulant. The unknown peak was determined to best match diglycolic acid. The development of a method for acetate is summaraized, and no significant amount of acetate was observed in the SRAT products tested. In addition, an alternative Gas Chromatograph (GC) method for glycolate is summarized.

Wiedenman, B. J.; White, T. L.; Mahannah, R. N.; Best, D. R.; Stone, M. E.; Click, D. R.; Lambert, D. P.; Coleman, C. J.




EPA Science Inventory

A standardized method for the analysis of perchlorate in plants was developed, based on dry weight, and applied to the analysis of plant organs, foodstuffs, and plant products. The procedure greatly reduced the ionic interferences in water extracts of plant materials. Ion chro...


Metallomics approach to trace element analysis in ustilago maydis using cellular fractionation, atomic absorption spectrometry, and size exclusion chromatography with ICP-MS detection.  


Huitlacoche is the ethnic name of the young fruiting bodies of Ustilago maydis, a common parasite of maize. In Mexico and other Latin American countries, this fungus has been traditionally appreciated as a local delicacy. In this work a metallomics approach was used with the determination of eight elements in huitlacoche by electrothermal atomic absorption spectrometry as one facet of this approach. The results obtained indicated relatively lower concentrations of commonly analyzed metals, as referred to the data reported for other mushroom types. This effect was ascribed to different accessibilities of elements, depending on fungus substrate (lower from plant than from soil). Subcellular fractionation was accomplished by centrifugation of cell homogenates suspended in Tris-HCl buffer. Recoveries of the fractionation procedure were in the range of 71-103%. For six elements (Cr, Cu, Fe, Mn, Ni, and Pb), the mean relative contributions in cytosol, cell walls, and mixed membrane fraction were 50.7, 48.2, and 1.1% respectively. To attain the molecular weight distribution of compounds containing target elements as an additional aspect of the metallomics approach, the fungus extract (1% sodium dodecyl sulfate in Tris-HCl, 30 mmol L(-)(1), pH 7.0) was analyzed by size exclusion chromatography with UV and ICP-MS detection. With spectrophotometric detection (280 nm), the elution of high molecular weight compounds was observed in the form of one peak (MW > 10 kDa), and several lower peaks appeared at higher retention times (MW < 10 kDa). On ICP-MS chromatograms, a coelution of (59)Co, (63)Cu, (57)Fe, (202)Hg, (60)Ni, and (80)Se with the first peak on the UV chromatogram was clearly observed, indicating that a fraction of each element incorporated with high molecular weight compounds (12.7, 19.8, 33.7, 100, 19.4, and 45.8%, respectively, based on the peak area measurements). From a comparison of (80)Se and (33)S chromatograms (for sulfur analysis, the extract was obtained in the absence of SDS), both elements coeluted with the first UV peak, but their lower molecular weight compounds were apparently different. These findings may contribute to a better understanding of the accumulation of elements in mushrooms. PMID:15969488

Muñoz, Alma Hortensia Serafin; Kubachka, Kevin; Wrobel, Kazimierz; Corona, Felix Gutierrez; Yathavakilla, Santha K V; Caruso, Joseph A; Wrobel, Katarzyna



Capability measurement of size-exclusion chromatography with a light-scattering detection method in a stability study of bevacizumab using the process capability indices.  


In this study, we investigated if the size-exclusion chromatography coupled with light-scattering and refractive index detection (SEC/LS/RI) method is fitted for its intended purpose and checked if the analytical method is able to provide enough conforming results. For this, the process capability indices Cp, Cpk, and Cpm were computed. The traditional X-chart and moving range (MR) chart were used by the same analyst to monitor the equipment in the laboratory over a 1-year period. For this, a bovine serum albumin (BSA) sample (0.3 mg mL(-1)) with a nominal Mw of 66.4 kDa was analyzed each working day. The results confirmed that the analytical method is in-control and stable. To determine whether the given process meets the present capability requirement and runs under the desired quality conditions, the Pearn and Shu (2003) method based on the lower confidence bound C on Cpm was used. The estimator Cpm was 1.81, and the lower confidence bound C was 1.40. We therefore conclude that the true value of the method capability Cpm is no less than 1.40 with a 95% level of confidence. This result indicates that the method is satisfactory and no stringent precision control is required. The usefulness of this method was applied in the characterization of bevacizumab commercial pharmaceutical preparations stored under different conditions that lead to aggregation. In this case, the computed Cpm index was 0.98 (0.70, 1.26), which indicates that the method does not comply with the specification limits and needs to be revised. The quality improvement effort should: (1) reduce the uncertainty in the absolute Mw determination; (2) either move the process mean closer to the target value or reduce the process variation, i.e. improve the method accuracy (?-T) and precision (?(2)). On this point, the Bayesian posterior distribution of the mean and standard deviation pointed out the need to control the precision but specially accuracy in order to reduce the overall uncertainty of analytical method and thus, the method is capable. PMID:24786652

Oliva, Alexis; Llabrés, Matías; Fariña, José B



Molecular weights and gyration radii of amylopectins determined by high-performance size-exclusion chromatography equipped with multi-angle laser-light scattering and refractive index detectors  

Microsoft Academic Search

High-performance size-exclusion chromatography (HPSEC) equipped with multi-angle laser-light scattering (MALLS) and refractive index (RI) detectors was used to determine weight-average molecular weight (Mw) and z-average radius of gyration (Rz) of amylopectin of selected starches. Ranges of Mw and Rz values of amylopectin were 7.0×107–5.7×109g\\/mol and 191–782nm, respectively. Amylopectins of waxy starches had substantially larger Mw than did those of normal

Sang-Ho Yoo; Jay-lin Jane



Molecular characterization of recombinant Hepatitis B surface antigen from Chinese hamster ovary and Hansenula polymorpha cells by high-performance size exclusion chromatography and multi-angle laser light scattering  

Microsoft Academic Search

The molecular weight and size of recombinant Hepatitis B surface antigen (HBsAg) derived from Chinese hamster ovary (CHO) and the Hansenula polymorph have been characterized by high-performance size exclusion chromatography with multi-angle laser light scattering (HPSEC-MALLS). The average molecular weight of CHO-derived HBsAg particle (CHO-rHBsAg) (4921kDa) was higher than that of H. polymorpha yeast strain (Hans-rHBsAg) (3010kDa). The size of

Weibin Zhou; Jingxiu Bi; Jan-Christer Janson; Yan Li; Yongdong Huang; Yan Zhang; Zhiguo Su



Miniaturized GC/MS instrumentation for in situ measurements: micro gas chromatography coupled with miniature quadrupole array and paul ion trap mass spectrometers  

NASA Technical Reports Server (NTRS)

Miniaturized chemical instrumentation is needed for in situ measurements in planetary exploration and other spaceflight applications where factors such as reduction in payload requirements and enhanced robustness are important. In response to this need, we are 'continuing to develop miniaturized GC/MS instrumentation which combines chemical separations by gas chromatography (GC) with mass spectrometry (MS) to provide positive identification of chemical compounds in complex mixtures of gases, such as those found in the International Space Station's cabin atmosphere. Our design approach utilizes micro gas chromatography components coupled with either a miniature quadrupole mass spectrometer array (QMSA) or compact, high-resolution Paul ion trap.

Holland, P.; Chutjian, A.; Darrach, M.; Orient, O.



Multiresidue determination of pesticides in malt beverages by capillary gas chromatography with mass spectrometry and selected ion monitoring.  


A method was developed to determine pesticides in malt beverages using solid phase extraction on a polymeric cartridge and sample cleanup with a MgSO4-topped aminopropyl cartridge, followed by capillary gas chromatography with electron impact mass spectrometry in the selected ion monitoring mode [GC-MS(SIM)]. Three GC injections were required to analyze and identify organophosphate, organohalogen, and organonitrogen pesticides. The pesticides were identified by the retention times of peaks of the target ion and qualifier-to-target ion ratios. GC detection limits for most of the pesticides were 5-10 ng/mL, and linearity was determined from 50 to 5000 ng/mL. Fortification studies were performed at 10 ng/mL for three malt beverages that differ in properties such as alcohol content, solids, and appearance. The recoveries from the three malt beverages were greater than 70% for 85 of the 142 pesticides (including isomers) studied. The data showed that the different malt beverage matrixes had no significant effect on the recoveries. This method was then applied to the screening and analysis of malt beverages for pesticides, resulting in the detection of the insectide carbaryl and the fungicide dimethomorph in real samples. The study indicates that pesticide levels in malt beverages are significantly lower than the tolerance levels set by the United States Environmental Protection Agency for malt beverage starting ingredients. The use of the extraction/cleanup procedure and analysis by GC-MS(SIM) proved effective in screening malt beverages for a wide variety of pesticides. PMID:15478993

Wong, Jon W; Webster, Michael G; Bezabeh, Dawit Z; Hengel, Mathew J; Ngim, Kenley K; Krynitsky, Alexander J; Ebeler, Susan E



Tailored Noise Waveform\\/ Collision-Induced Dissociation of Ions Stored in a Linear Ion Trap Combined with Liquid Chromatography\\/Fourier Transform Ion Cyclotron Resonance Mass Spectrometry  

Microsoft Academic Search

A new collision-induced dissociation (CID) technique based on broadband tailored noise waveform (TNW) excitation of ions stored in a linear ion trap has been developed. In comparison with the conventional sustained off-resonance irradiation (SORI) CID method commonly used in Fourier transform ion cyclotron resonance mass spectrometry, this MS\\/MS technique increases throughput by eliminating the long pump-down delay associated with gas

Andrey N. Vilkov; Bogdan Bogdanov; Liljiana Pasa-Tolic; David C. Prior; Gordon A. Anderson; Christophe D. Masselon; Ronald J. Moore; Richard D. Smith



Determination of ammonium in a buddingtonite sample by ion-chromatography  

USGS Publications Warehouse

An ion-chromatographic method for the direct determination of ammonium, potassium, and sodium in geologic materials is described. Samples are decomposed with a mixture of hydrofluoric and hydrochloric acids in a sealed polycarbonate bottle heated in a microwave oven. The ion-chromatograph separates the cations and determines them by conductivity measurement. The ammonium concentrations thus determined have been verified by use of an ammonia-specific electrode. A total of 32 analyses of ammonium salts by both techniques showed an average error of -4%, with a relative standard deviation (RSD) of 6%. The ammonium concentrations found in a buddingtonite sample had an RSD of 2.2% and their mean agreed with that obtained by the Kjeldahl method. By use of the prescribed dilution of the sample, detection limits of 0.1% can be achieved for all three cations. ?? 1986.

Klock, P.R.; Lamothe, P.J.



Ion size effects at ionic exclusion from dielectric interfaces and slit nanopores  

E-print Network

A previously developed field-theoretic model [R.D. Coalson et al., J. Chem. Phys. 102, 4584 (1995)] that treats core collisions and Coulomb interactions on the same footing is investigated in order to understand ion size effects on the partition of neutral and charged particles at planar interfaces and the ionic selectivity of slit nanopores. We introduce a variational scheme that can go beyond the mean-field (MF) regime and couple in a consistent way pore modified core interactions, steric effects, electrostatic solvation and image-charge forces, and surface charge induced electrostatic potential. We show that in the dilute limit, the MF and the variational theories agree well with MC simulation results, in contrast to a recent RPA method. The partition of charged Yukawa particles at a neutral dielectric interface (e.g air-water or protein-water interface) is investigated. It is shown that as a result of the competition between core collisions that push the ions towards the surface, and repulsive solvation and image forces that exclude them from the interface, a concentration peak of finite size ions sets in close to the dielectric interface. We also characterize the role played by the ion size on the ionic selectivity of neutral slit nanopores. We show that the complex interplay between electrostatic forces, excluded volume effects induced by core collisions and steric effects leads to an unexpected reversal in the ionic selectivity of the pore with varying pore size: while large pores exhibits a higher conductivity for large ions, narrow pores exclude large ions more efficiently than small ones.

Sahin Buyukdagli; C. V. Achim; T. Ala-Nissila



Gas chromatography/ion mobility spectrometry as a hyphenated technique for improved explosives detection and analysis  

NASA Technical Reports Server (NTRS)

Ion Mobility Spectrometry (IMS) is currently being successfully applied to the problem of on-line trace detection of plastic and other explosives in airports and other facilities. The methods of sample retrieval primarily consist of batch sampling for particulate residue on a filter card for introduction into the IMS. The sample is desorbed into the IMS using air as the carrier and negative ions of the explosives are detected, some as an adduct with a reagent ion such as Cl(-). Based on studies and tests conducted by different airport authorities, this method seems to work well for low vapor pressure explosives such as RDX and PETN, as well as TNT that are highly adsorptive and can be found in nanogram quantities on contaminated surfaces. Recently, the changing terrorist threat and the adoption of new marking agents for plastic explosives has meant that the sample introduction and analysis capabilities of the IMS must be enhanced in order to keep up with other detector developments. The IMS has sufficient analytical resolution for a few threat compounds but the IMS Plasmogram becomes increasingly more difficult to interpret when the sample mixture gets more complex.

Mercado, AL; Marsden, Paul



The analysis of aqueous mixtures using liquid chromatography-electrospray mass spectrometry  

SciTech Connect

The focus of this dissertation is the use of chromatographic methods coupled with electrospray mass spectrometry (ES-MS) for the determination of both organic and inorganic compounds in aqueous solutions. The combination of liquid chromatography (LC) methods and ES-MS offers one of the foremost methods for determining compounds in complex aqueous solutions. In this work, LC-ES-MS methods are devised using ion exclusion chromatography, reversed phase chromatography, and ion exchange chromatography, as well as capillary electrophoresis (CE). For an aqueous sample, these LC-ES-MS and CE-ES-MS techniques require no sample preparation or analyte derivatization, which makes it possible to observe a wide variety of analytes as they exist in solution. The majority of this work focuses on the use of LC-ES-MS for the determination of unknown products and intermediates formed during electrochemical incineration (ECI), an experimental waste remediation process. This report contains a general introduction to the project and the general conclusions. Four chapters have been removed for separate processing. Titles are: Chapter 2: Determination of small carboxylic acids by ion exclusion chromatography with electrospray mass spectrometry; Chapter 3: Electrochemical incineration of benzoquinone in aqueous media using a quaternary metal oxide electrode in the absence of a soluble supporting electrolyte; Chapter 4: The determination of electrochemical incineration products of 4-chlorophenol by liquid chromatography-electrospray mass spectrometry; and Chapter 5: Determination of small carboxylic acids by capillary electrophoresis with electrospray mass spectrometry.

Johnson, S.




EPA Science Inventory

Determination of carboxylic acids using non-suppressed conductivity and UV detections is described. The background conductance of 1-octanesulfonic acid, hexane sulfonic acid and sulfuric acid at varying concentrations was determined. Using 0.2 mM 1-octanesulfonic acid as a mobile...



EPA Science Inventory

Determination of carboxylic acids using non-suppressed conductivity and UV detections is described. he background conductance of I-octanesulfonic acid, hexane sulfonic acid and sulfuric acid at varying concentrations was determined. sing 0.2 MM I-octanesulfonic acid as a mobile p...


Development of high-throughput liquid chromatography injected ion mobility quadrupole time-of-flight techniques for analysis of complex peptide mixtures.  


The development of a multidimensional approach involving high-performance liquid chromatography (LC), ion mobility spectrometry (IMS) and tandem mass spectrometry is described for the analysis of complex peptide mixtures. In this approach, peptides are separated based on differences in their LC retention times and mobilities (as ions drift through He) prior to being introduced into a quadrupole/octopole/time-of-flight mass spectrometer. The initial LC separation and IMS dispersion of ions is used to label ions for subsequent fragmentation studies that are carried out for mixtures of ions. The approach is demonstrated by examining a mixture of peptides generated from tryptic digestion of 18 commercially available proteins. Current limitations of this initial study and potential advantages of the experimental approach are discussed. PMID:12458017

Lee, Young Jin; Hoaglund-Hyzera, Cherokee S; Srebalus Barnes, Catherine A; Hilderbrand, Amy E; Valentine, Stephen J; Clemmer, David E



Simultaneous determination of fatty, dicarboxylic and amino acids based on derivatization with isobutyl chloroformate followed by gas chromatography-positive ion chemical ionization mass spectrometry.  


Gas chromatography-mass spectrometry (GC-MS) with positive ion chemical ionization (PICI) using isobutane as reagent gas was applied for analysis of isobutoxycarbonyl/isobutyl derivatives of 13 fatty, 6 dicarboxylic and 13 amino acids in a single run. For all investigated compounds (except several amino acids) the quasimolecular ions [MH](+) were registered. Asparagine underwent fragmentation via decarboxylation followed by elimination of OC(4)H(9) ([M-117](+)), whereas serine and tyrosine produced the cluster ions [M+C(4)H(9)OCO](+). Estimated detection limits were 6-250 pg in the total ion current (TIC) mode and 3-10 times lower using the selected-ion monitoring (SIM) mode. PMID:14698242

Sobolevsky, Tim G; Revelsky, Alexander I; Revelsky, Igor A; Miller, Barbara; Oriedo, Vincent



Exclusive production of $?^0 ?^0$ pairs in ultrarelativistic heavy ion collisions  

E-print Network

We disuss exclusive electromagnetic production of two neutral $\\rho^0$~mesons and show the predictions for the $AA \\to AA \\rho^0 \\rho^0$ reactions for gold-gold collisions at the energy of $\\sqrt{s}$ = 200 GeV (RHIC) and for lead-lead collisions at the energy of $\\sqrt{s}$ = 5.5 TeV (LHC). The elementary cross section is calculated with the help of the vector-dominance-model (VDM)-Regge approach which usually very well describes the experimental data at large $\\gamma \\gamma$ energy. The low-energy $\\gamma \\gamma \\to \\rho^0 \\rho^0$ cross section is parametrized. The cross section for nuclear process is calculated by means of the equivalent photon approximation (EPA). We compare the results with realistic charge density with the results for monopole form factor.

Mariola Klusek-Gawenda; Antoni Szczurek



Removal of heavy metal ions from aquatic solutions by membrane chromatography  

Microsoft Academic Search

Polyvinylalcohol membranes were prepared by a solvent casting technique. Metal-complexation ligand, i.e. monochlorotriazinyl-dye Cibacron Blue F3GA was then attached. These membranes with a high water content of 119%, and containing 8.7 mmol Cibacron Blue F3GA\\/m2 were used in the adsorption\\/stripping of some selected heavy metal ions [Cu(II), Hg(II), Pb(II) and Cd(II)] from aquatic solutions containing varying initial concentration of metal

Adil Denizli; Ridvan Say; Yakup Arica



Semiautomated pH gradient ion-exchange chromatography of monoclonal antibody charge variants.  


A new approach using a chromatography system equipped with isocratic pumps and an electrolytic eluent generator (EG) is introduced, replacing external pH gradient delivery using conventional gradient systems, in which bottled buffers with preadjusted pH are mixed using a gradient pump. The EG is capable of generating high purity base or acid required for online preparation of the buffer at the point of use, utilizing deionized water as the only carrier stream. Typically, the buffer was generated from online titration of a reagent composed of low molecular weight amines. The reagent was delivered isocratically into a static mixing tee, where it was titrated to the required pH with electrolytically generated base or acid. The required pH gradient was thus conveniently generated by electrically controlling the concentration of titrant. Also, since the pH was adjusted at the point of use, this approach offered enhanced throughput in terms of eluent preparation time and labor, and with a more reproducible pH profile. The performance of the system was demonstrated by running pH gradients ranging from pH 8.2 to 10.9 on a polymer monolith cation-exchange column for high throughput profiling of charge heterogeneity of intact, basic therapeutic monoclonal antibodies. A high degree of flexibility in modulating the key parameters of the pH gradient, including the buffer concentration, the pH gradient slope and the operating pH range was demonstrated. This enabled fine-tuning of the separation conditions for each individual antibody in order to enhance the chromatographic resolution. PMID:25199803

Talebi, Mohammad; Shellie, Robert A; Hilder, Emily F; Lacher, Nathan A; Haddad, Paul R



Separation of polar betalain pigments from cacti fruits of Hylocereus polyrhizus by ion-pair high-speed countercurrent chromatography.  


Polar betacyanin pigments together with betaxanthins from ripe cactus fruits of Hylocereus polyrhizus (Cactaceae) were fractionated by means of preparative ion-pair high-speed countercurrent chromatography (IP-HSCCC) also using the elution-extrusion (EE) approach for a complete pigment recovery. HSCCC separations were operated in the classical 'head-to-tail' mode with an aqueous mobile phase. Different CCC solvent systems were evaluated in respect of influence and effectiveness of fractionation capabilities to separate the occurring pigment profile of H. polyrhizus. For that reason, the additions of two different volatile ion-pair forming perfluorinated carboxylic acids (PFCA) were investigated. For a direct comparison, five samples of Hylocereus pigment extract were run on preparative scale (900 mg) in 1-butanol-acetonitrile-aqueous TFA 0.7% (5:1:6, v/v/v) and the modified systems tert.-butyl methyl ether-1-butanol-acetonitrile-aqueous PFCA (2:2:1:5, v/v/v/v) using 0.7% and 1.0% trifluoroacetic acid (TFA) or heptafluorobutyric acid (HFBA) in the aqueous phase, respectively. The chemical affinity to the organic stationary CCC solvent phases and in consequence the retention of these highly polar betalain pigments was significantly increased by the use of the more lipophilic fluorinated ion-pair reagent HFBA instead of TFA. The HFBA additions separated more effectively the typical cacti pigments phyllocactin and hylocerenin from betanin as well as their iso-forms. Unfortunately, similar K(D) ratios and selectivity factors alpha around 1.0-1.1 in all tested solvent systems proved that the corresponding diastereomers, 15S-type pigments cannot be resolved from the 15R-epimers (iso-forms). Surprisingly, additions of the stronger ion-pair reagent (HFBA) resulted in a partial separation of hylocerenin from phyllocactin which were not resolved in the other solvent systems. The pigments were detected by means of HPLC-DAD and HPLC-electrospray ionization-MS using also authentic reference materials. PMID:19732900

Wybraniec, S?awomir; Stalica, Pawe?; Jerz, Gerold; Klose, Bettina; Gebers, Nadine; Winterhalter, Peter; Spórna, Aneta; Szaleniec, Maciej; Mizrahi, Yosef



Dissociative electron attachment negative ion mass spectrometry: a chlorine-specific detector for gas chromatography  

NASA Astrophysics Data System (ADS)

This work describes the application of negative ion chemical ionization, optimized for dissociative electron attachment (DEA), to location of unknown trace chlorinated compounds in complex gas chromatograms by selected ion recording (SIR) of m / z 35 and 37. The DEA-SIR technique is compared with other GC detectors, including the electron capture detector, electrolytic conductivity detector, the atomic emission detector and the chemical reaction interface mass spectrometry method, with respect to selectivity for chlorine, sensitivity, linear dynamic range, and general robustness and ease of use. When applied to quantitative analysis of target analytes such as polychlorobiphenyls, the DEA-SIR method has potential problems arising from the possibility of suppression effects due to abundant co-eluting components, and possible alleviating measures are discussed. In addition to these practical investigations, literature information on the fundamental physical and chemical phenomena underlying the DEA process is summarized in order to guide future work on extension to other compound types and on general improvements to the technique.

Curtis, Jonathan M.; Boyd, Robert K.



Determination of sulfonic acids and alkylsulfates by ion chromatography in water.  


A fast ion chromatographic method with suppressed conductivity detection has been developed for the simultaneous determination of methane-, ethane-, 1-propane-, 1-butane-, 1-pentane-, 1-hexane-, 1-heptane-, 1-octane-, 1-nonane-, 1-decane-, 1-dodecane-, dodecylbenzene-, p-toluene-, benzenesulfonic acids, octylsulfate and dodecylsulfate in water samples. Due to the high number of analytes and their heterogeneity, the effect of the mobile phase parameters (NaOH, CH(3)OH and CH(3)CN concentration) on the retention factors has been studied in detail, so achieving, for the first time, the separation among 15 of these analytes by a gradient elution. Detection limits included within 0.06-0.16 microM have been obtained. Interferences from Cl(-), NO(3)(-) and SO(4)(2-), possible anions present in water samples, have been considered and a SPE procedure has been developed for analytes enrichment and matrix removal in a seawater sample. This is the first application of an ion-exchange chromatographic method to a seawater sample for this kind of analytes. PMID:18585139

Bruzzoniti, Maria Concetta; De Carlo, Rosa Maria; Sarzanini, Corrado



Quantifying the labeling and the levels of plant cell wall precursors using ion chromatography tandem mass spectrometry.  


The biosynthesis of cell wall polymers involves enormous fluxes through central metabolism that are not fully delineated and whose regulation is poorly understood. We have established and validated a liquid chromatography tandem mass spectrometry method using multiple reaction monitoring mode to separate and quantify the levels of plant cell wall precursors. Target analytes were identified by their parent/daughter ions and retention times. The method allows the quantification of precursors at low picomole quantities with linear responses up to the nanomole quantity range. When applying the technique to Arabidopsis (Arabidopsis thaliana) T87 cell cultures, 16 hexose-phosphates (hexose-Ps) and nucleotide-sugars (NDP-sugars) involved in cell wall biosynthesis were separately quantified. Using hexose-P and NDP-sugar standards, we have shown that hot water extraction allows good recovery of the target metabolites (over 86%). This method is applicable to quantifying the levels of hexose-Ps and NDP-sugars in different plant tissues, such as Arabidopsis T87 cells in culture and fenugreek (Trigonella foenum-graecum) endosperm tissue, showing higher levels of galacto-mannan precursors in fenugreek endosperm. In Arabidopsis cells incubated with [U-(13)C(Fru)]sucrose, the method was used to track the labeling pattern in cell wall precursors. As the fragmentation of hexose-Ps and NDP-sugars results in high yields of [PO(3)](-)/or [H(2)PO(4)](-) ions, mass isotopomers can be quantified directly from the intensity of selected tandem mass spectrometry transitions. The ability to directly measure (13)C labeling in cell wall precursors makes possible metabolic flux analysis of cell wall biosynthesis based on dynamic labeling experiments. PMID:20442274

Alonso, Ana P; Piasecki, Rebecca J; Wang, Yan; LaClair, Russell W; Shachar-Hill, Yair



Potential of ion chromatography coupled to isotope ratio mass spectrometry via a liquid interface for beverages authentication.  


New tools for the determination of characteristic parameters for food authentication are requested to prevent food adulteration from which health concerns, unfair competition could follow. A new coupling in the area of compound-specific carbon 13 isotope ratio (?(13)C) analysis was developed to simultaneously quantify ?(13)C values of sugars and organic acids. The coupling of ion chromatography (IC) together with isotope ratio mass spectrometry (IRMS) can be achieved using a liquid interface allowing a chemical oxidation (co) of organic matter. Synthetic solutions containing 1 polyol (glycerol), 3 carbohydrates (sucrose, glucose and fructose) and 12 organic acids (gluconic, lactic, malic, tartaric, oxalic, fumaric, citric and isocitric) were used to optimize chromatographic conditions (concentration gradient and 3 types of column) and the studied isotopic range (-32.28 to -10.65‰) corresponds to the values found in food products. Optimum chromatographic conditions are found using an IonPac AS15, an elution flow rate of 0.3mLmin(-1) and a linear concentration gradient from 2 to 76mM (rate 21mMmin(-1)). Comparison between ?(13)C value individually obtained for each compound with the coupling IRMS and elemental analyzer, EA-IRMS, and the ones measured on the mixture of compounds by IC-co-IRMS does not reveal any isotope fractionation. Thus, under these experimental conditions, IC-co-IRMS results are accurate and reproducible. This new coupling was tested on two food matrices, an orange juice and a sweet wine. Some optimization is necessary as the concentration range between sugars and organic acids is too large: an increase in the filament intensity of the IRMS is necessary to simultaneously detect the two compound families. These first attempts confirm the good results obtained on synthetic solutions and the strong potential of the coupling IC-co-IRMS in food authentication area. PMID:24267317

Guyon, Francois; Gaillard, Laetitia; Brault, Audrey; Gaultier, Nicolas; Salagoïty, Marie-Hélène; Médina, Bernard



Separation of betalains from berries of Phytolacca americana by ion-pair high-speed counter-current chromatography.  


The first preparative fractionation of betalain pigments by means of ion-pair high-speed counter-current chromatography (IP-HSCCC) from berry extracts of Phytolacca americana (Phytolaccaceae) is presented. A novel HSCCC solvent system consisting of 1-butanol-acetonitrile-water (5:1:6, v/v/v) was applied using ion-pair forming trifluoroacetic acid at low concentration (0.7%, v/v). Affinity of polar betacyanins and betaxanthins to the organic stationary phase of the biphasic HSCCC solvent mixture was considerably improved. Partitioning coefficient values and influence of increasing trifluoroacetic acid additions to the biphasic solvent mixture were measured for all identified betacyanins and betaxanthins. Gentle separation by IP-HSCCC of the injected pigment extract (900 mg) yielded sufficient amounts of the principal pigments 15S-betanin/15R-isobetanin. The pure epimers separated by C18-HPLC were immediately studied by one- and two-dimensional NMR. In the recovered fractions, minor concentrated betacyanins and betaxanthins were significantly enriched by IP-HSCCC and were detected for the first time in the extracts of P. americana. IP-HSCCC and C18-HPLC were shown to be complementary techniques in the isolation procedure of recovering minor concentrated, highly polar and chemically instable betacyanins and betaxanthin from complex plant matrices. Altogether, identification of 17 betalains was achieved by HPLC-diode array detection-electrospray ionization MS/MS in the HSCCC fractions with their respective isomers, also resulting in the tentative elucidation of betacyanins with novel salicylic acid substitution pattern in the berry extracts of P. americana. PMID:18374932

Jerz, Gerold; Skotzki, Tanja; Fiege, Kathrin; Winterhalter, Peter; Wybraniec, S?awomir



Analysis of atmospheric aerosols by particle-induced X-ray emission, instrumental neutron activation analysis, and ion chromatography  

NASA Astrophysics Data System (ADS)

Particle-induced X-ray emission (PIXE), ion chromatography (IC), and occasionally also instrumental neutron activation analysis (INAA) were used in combination for the analysis of atmospheric aerosol samples that were collected on Nuclepore polycarbonate filters. A comparison of the results enabled us to evaluate the matrix effects (i.e., particle size effects) of the PIXE analysis for the light elements and to assess the water-solubility and/or speciation of a number of elements (e.g., S, K, Ca). Results are presented from several measurement campaigns at urban and forested sites in Europe, whereby PM10 or PM2.5 filter samples were taken. From the PIXE and IC results for a 2003 summer campaign at the K-puszta site in Hungary, it was estimated that organosulphates could be responsible for 20% of the total sulphur concentration and 30% of the organic aerosol in PM10. The comparison of the IC and PIXE data for K and Ca from the various sites indicated that most of the Ca was water-soluble (the mineral dust Ca was presumably mostly present as calcite, and perhaps also in part as gypsum); in contrast, for K, only half of it was typically water-soluble, indicating that it was to a large extent associated with insoluble mineral dust. Exceptions, with almost fully water-soluble K, were found for samples that were substantially impacted by biomass burning.

Maenhaut, Willy; Raes, Nico; Wang, Wan



Stability of plasma gamma-hydroxybutyrate determined by gas chromatography-positive ion chemical ionization-mass spectrometry.  


An effective method for the determination of gamma-hydroxybutyric acid (GHB) in human plasma is described that utilizes a simple liquid-liquid extraction procedure and gas chromatography-positive ion chemical ionization-mass spectrometry (GC-PCI-MS). The method has been used to study the stability of plasma GHB under several storage conditions. Following the extraction with acetonitrile, GHB and deuterated GHB (GHB-d(6)) were derivatized with N,O-bis[trimethylsilyl] trifluoroacetamide (BSFTA). After the separation on a capillary GC column, the derivatives were ionized with ammonia reagent gas and analyzed by MS. The lower limit of quantitation in 100 microL of plasma was 2.5 microg/mL, over a range from 2.5 to 250 microg/mL. The coefficients of variation did not exceed 3.9% and the mean measured concentrations did not deviate more than 8% from the target for both intra- and interassay precision and accuracy. Plasma GHB was found to be stable at -20 degrees C for up to 9 months, at room temperature for 48 h, and after 3 freeze/thaw cycles. It was also found to be stable in processed samples stored at room temperature for 5 days and for 15 days at -20 degrees C. PMID:14606997

Chen, Meng; Andrenyak, David M; Moody, David E; Foltz, Rodger L



Analysis of terpenoids from hemlock (Tsuga) species by solid-phase microextraction/gas chromatography/ion-trap mass spectrometry.  


A sampling method for determining the volatile terpenoid composition from single needles of seven Tsuga species was developed using headspace solid-phase microextraction (SPME). A reproducible sampling method for the volatile components was generated by examination of sample storage, method of needle cutting, and headspace sampling duration. Following SPME collection of the volatile compounds from the seven Tsuga species, gas chromatography/ion-trap mass spectrometry was used to identify 51 terpenoids present in the needle headspace. A semiquantitative method was devised to express individual terpenoid amounts as a percentage of all of the identified peaks in the chromatogram. The semiquantitative results permitted facile interspecies comparison using principal component analysis. Two components were able to account for 90% of the variance and were interpreted as a "species" component and a "resistance/susceptibility" component. Three interspecies groupings were evident from the principal component analysis: (1) Tsuga canadensis and Tsuga caroliniana; (2) Tsuga chinesnsis, Tsuga diversifolia, Tsuga heterophylla, and Tsuga sieboldii; and (3) Tsuga mertensiana. The finding that T. mertensiana was grouped alone and far removed from the other species adds to the morphological evidence that this species should be segregated from other Tsuga. PMID:12670144

Lagalante, Anthony F; Montgomery, Michael E



A method for analysis of wilfordmine in human plasma by liquid chromatography coupled with ion trap mass spectrometry.  


A simple and rapid liquid chromatography-ion trap mass spectrometric (LC-IT/MS) method has been developed and validated for quantification of wilfordmine in human plasma. After the protein precipitation was carried out by acetonitrile and the solution was cleaned by solid-phase extraction, the chromatographic separation was performed on a Zorbax Plus RRHD C18 column by using a mixture of acetonitrile and 10.0 mmol/L ammonium acetate solution (70:30, v/v) as the mobile phase at a flow rate of 0.7 mL/min. Detection was performed on an atmospheric-pressure chemical ionization source in the positive multiple reaction monitoring mode using aconitine as an internal standard (IS) with transitions of m/z 806?710 for wilfordmine, and 646?586 for IS, respectively. The obtained calibration curve was linear (r = 0.9992) over the concentration range of 0.5-100.0 ?g/L with a lower limit of quantification of 0.5 ?g/L in plasma. The intra- and interday relative standard deviations were <7.0 and 12.3%, respectively. The recoveries were between 86.0 and 97.0%. The proposed method was found to be applicable to clinical studies. PMID:24895446

Zhou, Lixin; Zhu, Hao; Ouyang, Xiaokun; Zhao, Jinshun; Chen, Xiaohong



Fast carbohydrate analysis via liquid chromatography coupled with ultra violet and electrospray ionization ion trap detection in 96-well format.  


A fast carbohydrate screening platform processible in 96-well format is described. The method is suitable for the determination of various carbohydrates out of complex mixtures as obtained by acidic hydrolysis of carbohydrates polymers. The chromatographic conditions for an efficient separation (12min) and the derivatization process with 1-phenyl-3-methyl-5-pyrazolone (PMP) were optimized for high resolution separation and simultaneous determination of deoxy-, amino-, anhydro-sugars as well as hexoses, pentoses, dimers, uronic acids and degradation products like furfural and hydroxymethylfurfural (HMF). The potential to quantify with UV- and MS-detector in the same range has been demonstrated for 20 different compounds. Finally, the matrix effects of the hydrolysis were positively evaluated. The micro scale hydrolysis and PMP-derivatization without any extraction or drying steps, both in 96-well format, result in a fast and intuitive sample preparation. In combination with a fast liquid chromatography coupled to UV and electrospray ionization ion trap detection (LC-UV-ESI-MS/MS) for the qualification and quantification of various sugars, dimers and degradation products, this method shows great performance in carbohydrate analysis. PMID:24861788

Rühmann, Broder; Schmid, Jochen; Sieber, Volker



Determination of arsenic speciation in sulfidic waters by Ion Chromatography Hydride-Generation Atomic Fluorescence Spectrometry (IC-HG-AFS).  


A method for the analysis of arsenic species in aqueous sulfide samples is presented. The method uses an ion chromatography system connected with a Hydride-Generation Atomic Fluorescence Spectrometer (IC-HG-AFS). With this method inorganic As(III) and As(V) species in water samples can be analyzed, including arsenite (HnAs(III)O3(n-3)), thioarsenite (HnAs(III)S3(n-3)), arsenate (HnAs(V)O4(n-3)), monothioarsenate (HnAs(V)SO3(n-3)), dithioarsenate (HnAs(V)S2O2(n-3)), trithioarsenate (HnAs(V)S3O(n-3)) and tetrathioarsenate (HnAs(V)S4(n-3)). The peak identification and retention times were determined based on standard analysis of the various arsenic compounds. The analytical detection limit was ~1-3 µg L(-1) (LOD), depending on the quality of the baseline. This low detection limit makes this method also applicable to discriminate between waters meeting the drinking water standard of max. 10 µg L(-1) As, and waters that do not meet this standard. The new method was successfully applied for on-site determination of arsenic species in natural sulfidic waters, in which seven species were unambiguously identified. PMID:25059187

Keller, Nicole S; Stefánsson, Andri; Sigfússon, Bergur



Characterization of metabolites of sibutramine in primary cultures of rat hepatocytes by liquid chromatography-ion trap mass spectrometry.  


Liquid chromatography-ion trap mass spectrometry was used for the detection and structural characterization of metabolites of the anti-obesity drug sibutramine. Metabolites were profiled from incubations of sibutramine in primary cultures of rat hepatocytes. In addition, enantioselectivity of sibutramine metabolism was investigated by carrying out separate incubations with (R)- and (S)-sibutramine. As a result, biotransformation profile for sibutramine with rat hepatocytes is proposed. Nineteen metabolites and several of their isomers formed via demethylation, hydroxylation, dehydrogenation, acetylation, attachment of CO(2), and glucuronidation were identified in MS(2) and MS(3) experiments, though the exact position of the functionality, mostly hydroxylation, could not always be determined from the mass spectrometric information. However, clear enantioselective formation was observed for two hydroxyl derivatives and two glucuronide conjugates, indicating that the hydroxyl/glucuronic acid moiety in those structures is close to the chiral center. Most of the metabolites found in this study are new metabolites of sibutramine, which were not previously reported. PMID:19066859

Hakala, Kati S; Link, Marek; Szotakova, Barbora; Skalova, Lenka; Kostiainen, Risto; Ketola, Raimo A



Amperometric detection studies of Nafion/indium hexacyanoferrate film for the determination of electroinactive cations in ion chromatography.  


An amperometric detector based on the chemical modification of Nafion and indium (III) hexacyanoferrate (II, III) thin film (Nafion/In-CN-Fe) onto a glassy carbon (GC) electrode, was first successfully used for the determination of electroinactive cations (Li+, Na+, K+, Rb+, Cs+, NH4+) in single column ion chromatography (IC). A set of well-defined peaks of electroinactive cations was obtained. The detection limits of the cations are 8.9 x 10(-6) mol/L for Li+, 2.3 x 10(-6) mol/L for Na+, 5.2 x 10(-6) mol/L for K+, 4.8 x 10(-6) mol/L for Rb+, 4.0 x 10(-6) mol/L for Cs+ and 5.3 x 10(-6) mol/L for NH4+ at a single-to-noise ratio of 3. The proposed method was quick, sensitive and simple. The cations in rainwater and mineral water were successfully analyzed by this method. PMID:11227451

Xu, Q; Zhang, S; Zhang, W; Jin, L; Tanaka, K; Haraguchi, H; Itoh, A



Analytical characterization of complex, biotechnological feedstocks by pH gradient ion exchange chromatography for purification process development.  


The accelerating growth of the market for proteins and the growing interest in new, more complex molecules are bringing new challenges to the downstream process development of these proteins. This results in a demand for faster, more cost efficient, and highly understood downstream processes. Screening procedures based on high-throughput methods are widely applied nowadays to develop purification processes for proteins. However, screening highly complex biotechnological feedstocks, such as complete cell lysates containing target proteins often expressed with a low titre, is still very challenging. In this work we demonstrate a multidimensional, analytical screening approach based on pH gradient ion exchange chromatography (IEC), gel electrophoresis and protein identification via mass spectrometry to rationally characterize a biotechnological feedstock for the purpose of purification process development. With this very simple characterization strategy a two-step purification based on consecutive IEC operations was rapidly laid out for the purification of a diagnostic protein from a cell lysate reaching a purity of ?80%. The target protein was recombinantly produced using an insect cell expression system. PMID:24016717

Kröner, Frieder; Hanke, Alexander T; Nfor, Beckley K; Pinkse, Martijn W H; Verhaert, Peter D E M; Ottens, Marcel; Hubbuch, Jürgen



Control of impurities in diphenhydramine hydrochloride by an ion-pairing, reverse liquid chromatography method.  


A precise, sensitive, repeatable and robust reverse-phase liquid chromatographic method has been developed for the control of seven possible impurities of diphenhydramine hydrochloride. The robustness of the method was examined by varying, in turn, each of four mobile-phase parameters (acetonitrile content, buffer salt concentration, ion-pair reagent concentration and pH). The method was linear in the range 0-0.14 mg mL(-1) for diphenhydramine hydrochloride with an acceptable precision and accuracy, and a limit of detection of 0.17 microg mL(-1). Five samples of diphenhydramine hydrochloride from two sources were analysed with the developed liquid chromatographic method. PMID:11291747

Henderson, L; Miller, J H; Skellern, G G



Automated determination of bromide in waters by ion chromatography with an amperometric detector  

USGS Publications Warehouse

An automated ion chromatograph, including a program controller, an automatic sampler, an integrator, and an amperometric detector, was used to develop a procedure for the determination of bromide in rain water and many ground waters. Approximately 10 min is required to obtain a chromatogram. The detection limit for bromide is 0.01 mg l-1 and the relative standard deivation is <5% for bromide concentrations between 0.05 and 0.5 mg l-1. Chloride interferes if the chloride-to-bromide ratio is greater than 1 000:1 for a range of 0.01-0.1 mg l-1 bromide; similarly, chloride interferes in the 0.1-1.0 mg l-1 range if the ratio is greater than 5 000:1. In the latter case, a maximum of 2 000 mg l-1 of chloride can be tolerated. Recoveries of known concentrations of bromide added to several samples, ranged from 97 to 110%. ?? 1983.

Pyen, G.S.; Erdmann, D.E.



Measurement of trace nitrate concentrations in seawater by ion chromatography with valve switching  

NASA Astrophysics Data System (ADS)

An ion chromatographic method with a valve switching facility was developed to determine trace nitrate concentrations in seawater using two pumps, two different suppressors, and two columns. A carbohydrate membrane desalter was used to reduce the high concentrations of sodium salts in samples. In this method, trace nitrate was eluted from the concentrator column to the analytical columns, while the matrix fl owed to waste. Neither chemical pre-treatment nor sample dilution was required. In the optimized separation conditions, the method showed good linearity ( R >0.99) in the 0.05 and 50 mg/L concentration range, and satisfactory repeatability (RSD<5%, n =6). The limit of detection for nitrate was 0.02 mg/L. Results showed that the valve switching system was suitable and practical for the determination of trace nitrate in seawater.

Du, Juan; Fa, Yun; Zheng, Yue; Li, Xuebing; Du, Fanglin; Yang, Haiyan



Ion-pair chromatography for simultaneous analysis of ethionamide and pyrazinamide from their porous microparticles.  


Ethionamide (ETA) and pyrazinamide (PZA) are considered the drugs of choice for the treatment of multidrug-resistant tuberculosis. Current methods available in the literature for simultaneous determination of ETA and PZA have low sensitivity or involve column modifications with lipophilic cations. The aim of this study was to develop a simple and validated reversed-phase ion-pair HPLC method for simultaneous determination of ETA and PZA for the characterization of polymeric-based porous inhalable microparticles in in vitro and spiked human serum samples. Chromatographic separation was achieved on a Phenomenex C18 column (250 mm?×?4.6 mm) using a Shimadzu LC 10 series HPLC. The mobile phase consisted of A: 0.01% trifluoroacetic acid in distilled water and B: ACN/MeOH at 1:1 v/v. Gradient elution was run at a flow rate of 1.5 mL/min and a fixed UV wavelength of 280 nm. The validation characteristics included accuracy, precision, linearity, analytical range, and specificity. Calibration curves at seven levels for ETA and PZA were linear in the analytical range of 0.1-3.0 ?g/mL with correlation coefficient of r (2)?>?0.999. Accuracy for both ETA and PZA ranged from 94 to 106% at all quality control (QC) standards. The method was precise with relative standard deviation less than 2% at all QC levels. Limits of quantitation for ETA and PZA were 50 and 70 ng/mL, respectively. There was no interference from either the polymeric matrix ions or the biological matrix in the analysis of ETA and PZA. PMID:23990078

Bhanushali, Chintan J; Zidan, Ahmed S; Rahman, Ziyaur; Habib, Muhammad J



Determination of trace concentrations of bromate in municipal and bottled drinking waters using a hydroxide-selective column with ion chromatography  

Microsoft Academic Search

The International Agency for Research on Cancer determined that bromate is a potential human carcinogen, even at low ?g\\/l levels in drinking water. Bromate is commonly produced from the ozonation of source water containing naturally occurring bromide. Traditionally, trace concentrations of bromate and other oxyhalides in environmental waters have been determined by anion exchange chromatography with an IonPac AS9-HC column

Brian M. De Borba; Jeff S. Rohrer; Chris A. Pohl; Charanjit Saini



Analysis of phospholipid species in human blood using normal-phase liquid chromatography coupled with electrospray ionization ion-trap tandem mass spectrometry  

Microsoft Academic Search

A narrow-bore normal-phase high-performance liquid chromatography (HPLC) method was developed for separation of phospholipid classes in human blood. The separation was obtained using an HPLC diol column and a gradient of chloroform and methanol with 0.1% formic acid, titrated to pH 5.3 with ammonia and added 0.05% triethylamine. The HPLC system was coupled on-line with an electrospray ionisation ion-trap mass

Steinar Uran; Åsmund Larsen; Petter Balke Jacobsen; Tore Skotland



Determination of phenolic preservatives in gelatin and vacant capsules for medicine use by ion-suppression reversed-phase high performance liquid chromatography  

Microsoft Academic Search

A reliable method for the simultaneous determination of phenolic preservatives parahydroxybenzoic acid, methyl, ethyl, propyl, butyl and pentyl parahydroxybenzoates in gelatin and vacant capsules for medical use has been developed by ion-suppression reversed-phase high performance liquid chromatography. Separation was carried out on a Kromasil C18 column by isocratic elution using methanol–perchloric acid (pH 2.0; 10mM) (60:40, v\\/v) at a flow-rate

Hongzhen Lian; Danli Li; Xiaoxian Wu; Liching Tian



Automated LC–LC–MS–MS platform using binary ion-exchange and gradient reversed-phase chromatography for improved proteomic analyses  

Microsoft Academic Search

A simple multidimensional liquid chromatography system utilizing an isocratic pump and a HPLC system is described for the comprehensive proteomic analysis of complex peptide digest mixtures by coupled LC–LC–MS–MS techniques. A binary ion-exchange separation was achieved through the use of a strong cation-exchange column followed by a reversed-phase column for data-dependent LC–MS–MS analysis of the unbound analytes, and following salt

Michael T. Davis; Jill Beierle; Edward T. Bures; Michael D. McGinley; Jessica Mort; John H. Robinson; Chris S. Spahr; Wen Yu; Roland Luethy; Scott D. Patterson



Use of an ion-pairing reagent for high-performance liquid chromatography–atmospheric pressure chemical ionization mass spectrometry determination of anionic anticoagulant rodenticides in body fluids  

Microsoft Academic Search

The on-line combination of high-performance liquid chromatography with mass spectrometry (HPLC–MS) has become a powerful tool for trace analysis thanks to the developments in interface techniques. However, non-volatile salts such as ion-pairing reagents are considered to be incompatible with HPLC–MS systems; they cause drops in analyte signals because of contamination of mass analyzers and also because of blocking of the

Fuyu Guan; Akira Ishii; Hiroshi Seno; Kanako Watanabe-Suzuki; Takeshi Kumazawa; Osamu Suzuki



Monitoring Wadi El Raiyan lakes of the egyptian desert for inorganic pollutants by ion-selective electrodes, ion chromatography, and inductively coupled plasma spectroscopy.  


Wadi El Raiyan is a great depression located southwest of Cairo in the western desert of Egypt, one of the most arid regions of the world. In 1973, Wadi El Raiyan was connected with the agricultural wastewater drainage system of the El Faiyum province to provide a reservoir for the wastewater that exceeded the capacity of Lake Qarun north of the province. Pollutants from agricultural waste including pesticides and fertilizers as well as other effluents of industrial activities and runoffs certainly will pass into the biotic elements of the ecosystem. This report presents the status of inorganic pollutants including anions, cations, and trace metals in the two lakes and the surrounding springs of Wadi El Raiyan using ion chromatography, ion-selective electrodes, and inductively coupled plasma emission spectroscopy. The report also includes the levels of selected metals in the vegetation community of the area. The result of this investigation revealed a great improvement in water quality of the Wadi El Raiyan lakes compared to 1988 report by Saleh et al. Mercury was not detected in any of the samples and the level of lead was significantly reduced. Cadmium levels were much higher than those seen earlier. The higher level of cadmium might be used as an indicator to track the contamination of water by human waste. Concentrations of common anions were not significantly different from those reported earlier. However, an increase in the level of cyanide was observed. Levels of heavy metals in vegetation around the lakes were also found to be lower than previously reported. PMID:10702352

Saleh, M A; Ewane, E; Jones, J; Wilson, B L



Rapid and direct determination of iodide in seawater by electrostatic ion chromatography.  


An electrostatic ion chromatographic (IC) method for rapid and direct determination of iodide in seawater is reported. Separation was achieved using a reversed-phase ODS packed column (250x4.6 mm I.D.) modified by coating with Zwittergent-3-14 micelles, with an eluent comprising an aqueous solution containing 0.2 mM NaClO4 and 0.3 mM Zwittergent-3-14 and using UV detection at 210 nm. Samples prepared by dissolving NaIO3, NaNO2, NaBr, NaBrO3, NaNO3, NaI, and NaSCN in artificial or real seawaters were analyzed using this IC system. Nitrite, iodate, bromide, bromate, and nitrate showed very little or no retention, while iodide and thiocyanate were well separated, being eluted within 6 and 16 min, respectively. The detection limit for iodide obtained by injecting 400 microL of sample was 0.011 microM (S/N = 3), and the precision values obtained by analyzing samples containing 0.1 or 0.3 microM iodide in real seawater samples were 2.3% RSD and 1.2% RSD, respectively. Direct determination of iodide in real seawater samples was possible using this proposed IC system. PMID:12108640

Hu, Wenzhi; Yang, Pei-Jie; Hasebe, Kiyoshi; Haddad, Paul R; Tanaka, Kazuhiko



Speciation of arsenic in different types of nuts by ion chromatography-inductively coupled plasma mass spectrometry.  


In this work the quantitative determination and analytical speciation of arsenic were undertaken in different types of nuts, randomly purchased from local markets. The hardness of the whole nuts and high lipid content made the preparation of this material difficult for analysis. The lack of sample homogeneity caused irreproducible results. To improve the precision of analysis, arsenic was determined separately in nut oil and in the defatted sample. The lipids were extracted from the ground sample with the two portions of a mixture of chloroform and methanol (2:1). The defatted material was dried and ground again, yielding a fine powder. The nut oil was obtained by combining the two organic extracts and by evaporating the solvents. The two nut fractions were microwave digested, and total arsenic was determined by inductively coupled plasma mass spectrometry (ICP-MS). The results obtained for oils from different types of nuts showed element concentration in the range 2.9-16.9 ng g(-)(1). Lower levels of arsenic were found in defatted material (<0.1 ng g(-)(1) with the exception of Brazil nuts purchased with and without shells, 3.0 and 2.8 ng g(-)(1) respectively). For speciation analysis of arsenic in nut oils, elemental species were extracted from 2 g of oil with 12 mL of chloroform/methanol (2:1) and 8 mL of deionized water. The aqueous layer, containing polar arsenic species, was evaporated and the residue dissolved and analyzed by ion chromatography-ICP-MS. The anion exchange chromatography enabled separation of As(III), dimethylarsinic acid (DMAs(V)), monomethylarsonic acid (MMAs(V)), and As(V) within 8 min. Several types of nuts were analyzed, including walnuts, Brazil nuts, almonds, cashews, pine nuts, peanuts, pistachio nuts, and sunflower seeds. The recovery for the speciation procedure was in the range 72.7-90.6%. The primary species found in the oil extracts were As(III) and As(V). The arsenic concentration levels in these two species were 0.7-12.7 and 0.5-4.3 ng g(-)(1), respectively. The contribution of As in DMAs(V) ranged from 0.1 +/- 0.1 ng g(-)(1) in walnuts to 1.3 +/- 0.3 ng g(-)(1) in pine nuts. MMAs(V) was not detected in almonds, peanuts, pine nuts, sunflower seeds, or walnuts, and the highest concentration was found in pistachio nuts (0.5 +/- 0.2 ng g(-)(1)). PMID:15030196

Kannamkumarath, Sasi S; Wróbel, Kazimierz; Wróbel, Katarzyna; Caruso, Joseph A



Chromatography Theory and Modes  

NSDL National Science Digital Library

This site is an introductory lecture in outline form about chromatography. It appears directed toward biochemical separations. The coverage is broad but not deep. Discussions of counter-current separations, gel permeation, and ion exchange are accompanied by some mention of partition, paper, and gas chromatography. Useful illustrations are also included.


Qualitative and quantitative characterization of plasma proteins when incorporating traveling wave ion mobility into a liquid chromatography-mass spectrometry workflow for biomarker discovery: use of product ion quantitation as an alternative data analysis tool for label free quantitation.  


Discovery of protein biomarkers in clinical samples necessitates significant prefractionation prior to liquid chromatography-mass spectrometry (LC-MS) analysis. Integrating traveling wave ion mobility spectrometry (TWIMS) enables in-line gas phase separation which when coupled with nanoflow liquid chromatography and data independent acquisition tandem mass spectrometry, confers significant advantages to the discovery of protein biomarkers by improving separation and inherent sensitivity. Incorporation of TWIMS leads to a packet of concentrated ions which ultimately provides a significant improvement in sensitivity. As a consequence of ion packeting, when present at high concentrations, accurate quantitation of proteins can be affected due to detector saturation effects. Human plasma was analyzed in triplicate using liquid-chromatography data independent acquisition mass spectrometry (LC-DIA-MS) and using liquid-chromatography ion-mobility data independent acquisition mass spectrometry (LC-IM-DIA-MS). The inclusion of TWIMS was assessed for the effect on sample throughput, data integrity, confidence of protein and peptide identification, and dynamic range. The number of identified proteins is significantly increased by an average of 84% while both the precursor and product mass accuracies are maintained between the modalities. Sample dynamic range is also maintained while quantitation is achieved for all but the most abundant proteins by incorporating a novel data interpretation method that allows accurate quantitation to occur. This additional separation is all achieved within a workflow with no discernible deleterious effect on throughput. Consequently, TWIMS greatly enhances proteome coverage and can be reliably used for quantification when using an alternative product ion quantification strategy. Using TWIMS in biomarker discovery in human plasma is thus recommended. PMID:24397486

Daly, Charlotte E; Ng, Leong L; Hakimi, Amirmansoor; Willingale, Richard; Jones, Donald J L



Qualitative and Quantitative Characterization of Plasma Proteins When Incorporating Traveling Wave Ion Mobility into a Liquid Chromatography–Mass Spectrometry Workflow for Biomarker Discovery: Use of Product Ion Quantitation As an Alternative Data Analysis Tool for Label Free Quantitation  

PubMed Central

Discovery of protein biomarkers in clinical samples necessitates significant prefractionation prior to liquid chromatography–mass spectrometry (LC–MS) analysis. Integrating traveling wave ion mobility spectrometry (TWIMS) enables in-line gas phase separation which when coupled with nanoflow liquid chromatography and data independent acquisition tandem mass spectrometry, confers significant advantages to the discovery of protein biomarkers by improving separation and inherent sensitivity. Incorporation of TWIMS leads to a packet of concentrated ions which ultimately provides a significant improvement in sensitivity. As a consequence of ion packeting, when present at high concentrations, accurate quantitation of proteins can be affected due to detector saturation effects. Human plasma was analyzed in triplicate using liquid-chromatography data independent acquisition mass spectrometry (LC-DIA-MS) and using liquid-chromatography ion-mobility data independent acquisition mass spectrometry (LC-IM-DIA-MS). The inclusion of TWIMS was assessed for the effect on sample throughput, data integrity, confidence of protein and peptide identification, and dynamic range. The number of identified proteins is significantly increased by an average of 84% while both the precursor and product mass accuracies are maintained between the modalities. Sample dynamic range is also maintained while quantitation is achieved for all but the most abundant proteins by incorporating a novel data interpretation method that allows accurate quantitation to occur. This additional separation is all achieved within a workflow with no discernible deleterious effect on throughput. Consequently, TWIMS greatly enhances proteome coverage and can be reliably used for quantification when using an alternative product ion quantification strategy. Using TWIMS in biomarker discovery in human plasma is thus recommended. PMID:24397486



The mechanism for the exclusion of sugars from the water in a model of the liveing cell: the ion-exchange resin: pore size or water structure?  


The equilibrium distribution coefficients (p-value) of D-arabinose between the water in sulfonate ion-exchange resin and the external aqueous solution vary with the nature of the five alkali metal counterions studied. The strongest exclusion (lowest p-value) is found in the Li+ resin and the least exclusion (highest p-value) in the Cs+ resin. The p-value decreases with the increasing atomic weights for the alkali-metal ions: pCs+ greater than or equal to pRb+ greater than or equal to pRb+ greater than or equal to pK+ greater than pNa+ greater than pLi+. The water contents of these resins, on the other hand, vary in the opposite direction, being highest for the Li+ resin and lowest for the Cs+ resin. These data disprove the pore size theory but fully substantiate the predictions of the association-induction hypothesis. PMID:1197383

Ling, G N; Sobel, A M



Ultrafiltration by a compacted clay membrane-II. Sodium ion exclusion at various ionic strengths  

USGS Publications Warehouse

Several recent laboratory studies and field investigations have indicated that shales and compacted clay minerals behave as semipermeable membranes. One of the properties of semipermeable membranes is to retard or prevent the passage of charged ionic species through the membrane pores while allowing relatively free movement of uncharged species. This phenomenon is termed salt filtering, reverse osmosis, or ultrafiltration. This paper shows how one can proceed from the ion exchange capacity of clay minerals and, by means of Donnan membrane equilibrium concept and the Teorell-Meyer-Siever theory, develop a theory to explain why and to what extent ultrafiltration occurs when solutions of known concentration are forced to flow through a clay membrane. Reasonable agreement between theory and laboratory results were found. The concentration of the ultrafiltrate was always greater than predicted because of uncertainty in values of some parameters in the equations. Ultrafiltration phenomena may be responsible for the formation of some subsurface brines and mineral deposits. The effect should also be taken into consideration in any proposal for subsurface waste emplacement in an environment containing large quantities of clay minerals. ?? 1973.

Hanshaw, B.B.; Coplen, T.B.



Development and validation of an ion-exchange chromatography method for heparin and its impurities in heparin products.  


An anion-exchange liquid chromatography method for the determination of heparin and its impurities (dermatan sulfate and oversulfated chondroitin sulfate) was developed using chemometric-assisted optimization, including multivariate experimental design and response surface methodology. The separation of heparin, dermatan sulfate, and oversulfated chondroitin sulfate (Rs above 2.0) was achieved on a Dionex RF IC IonPac AS22 column with a gradient elution of 10-70% of 2.5 M sodium chloride and 20 mM Tris phosphate buffer (pH 2.1) at a flow rate of 0.6 mL/min and UV detection at 215 nm. Method validation shows good linearity (r > 0.99), acceptable precision (%relative standard deviations <11.4%) and trueness (%recovery of 92.3-103.9%) for all analytes. The limits of detection for dermatan sulfate and oversulfated chondroitin sulfate are equivalent to 0.11% w/w (10.5 ?g/mL) and 0.07% w/w (7.2 ?g/mL), while the limits of quantification are 0.32% w/w (31.5 ?g/mL) and 0.22% w/w (22.0 ?g/mL) relative to heparin, respectively. The method is specific for heparin, dermatan sulfate, and oversulfated chondroitin sulfate without interference from mobile phase and sample matrices and could be used for accurate quantitation the drug and its impurities in a single run. Applications of the method reveal contents of heparin between 90.3 and 97.8%. Dermatan sulfate and oversulfated chondroitin sulfate were not detected in any of the real-life samples. PMID:25146711

Thiangthum, Sumate; Heyden, Yvan Vander; Buchberger, Wolfgang; Viaene, Johan; Prutthiwanasan, Brompoj; Suntornsuk, Leena



Chemometric Analysis of Gas Chromatography – Mass Spectrometry Data using Fast Retention Time Alignment via a Total Ion Current Shift Function  

SciTech Connect

A critical comparison of methods for correcting severely retention time shifted gas chromatography-mass spectrometry (GC-MS) data is presented. The method reported herein is an adaptation to the Piecewise Alignment Algorithm to quickly align severely shifted one-dimensional (1D) total ion current (TIC) data, then applying these shifts to broadly align all mass channels throughout the separation, referred to as a TIC shift function (SF). The maximum shift varied from (-) 5 s in the beginning of the chromatographic separation to (+) 20 s toward the end of the separation, equivalent to a maximum shift of over 5 peak widths. Implementing the TIC shift function (TIC SF) prior to Fisher Ratio (F-Ratio) feature selection and then principal component analysis (PCA) was found to be a viable approach to classify complex chromatograms, that in this study were obtained from GC-MS separations of three gasoline samples serving as complex test mixtures, referred to as types C, M and S. The reported alignment algorithm via the TIC SF approach corrects for large dynamic shifting in the data as well as subtle peak-to-peak shifts. The benefits of the overall TIC SF alignment and feature selection approach were quantified using the degree-of-class separation (DCS) metric of the PCA scores plots using the type C and M samples, since they were the most similar, and thus the most challenging samples to properly classify. The DCS values showed an increase from an initial value of essentially zero for the unaligned GC-TIC data to a value of 7.9 following alignment; however, the DCS was unchanged by feature selection using F-Ratios for the GC-TIC data. The full mass spectral data provided an increase to a final DCS of 13.7 after alignment and two-dimensional (2D) F-Ratio feature selection.

Nadeau, Jeremy S.; Wright, Bob W.; Synovec, Robert E.



Simultaneous determination of total nitrogen and total phosphorus in environmental waters using alkaline persulfate digestion and ion chromatography.  


An ion chromatography (IC) method was developed for the simultaneous determination of total nitrogen and total phosphorus after alkaline persulfate digestion. This study takes advantage of advances in construction of high-resolution, high-capacity anion-exchange columns that can better tolerate the matrices typically encountered when a determination of total nitrogen and total phosphorous is required. Here, we used an electrolytically generated hydroxide eluent combined with a high-capacity, hydroxide-selective, anion-exchange column for the determination of total nitrogen (as nitrate-N) and total phosphorus (as phosphate-P) in environmental samples by IC. This method yielded LODs for nitrate-N and phosphate-P of 1.0 and 1.3 ?g/L, respectively. The LOQs determined for these analytes were 3.4 and 4.2 ?g/L, respectively. Due to the dilution factor required and the blank nitrate-N concentration after the persulfate digestion, the quantification limits increased for nitrate-N and phosphate-P to 171 and 63 ?g/L, respectively. The suitability of the method was evaluated by determining the nitrogen and phosphorus concentrations from known concentrations of organic-containing nitrogen and phosphorus compounds. In addition, environmental samples consisting of six different wastewaters and 48 reservoir samples were evaluated for total nitrogen and phosphorus. The recoveries of nitrogen and phosphorus from the organic-containing compounds ranged from 93.1 to 100.1% and 85.2 to 97.1%, respectively. In addition, good correlation between results obtained by the colorimetric method and IC was also observed. The linearity, accuracy, and evaluation of potential interferences for determining TN and TP will be discussed. PMID:25441080

De Borba, Brian M; Jack, Richard F; Rohrer, Jeffrey S; Wirt, Joan; Wang, Dongmei



Characterization of metabolites in rat plasma after intravenous administration of salvianolic acid A by liquid chromatography/time-of-flight mass spectrometry and liquid chromatography/ion trap mass spectrometry.  


Salvianolic acid A (SalA) is one of the main active constituents in Salvia miltiorrhiza (Danshen). Although the pharmacokinetics of SalA in rats after intravenous (i.v.) administration of Danshen injection has been reported, the information relevant to the metabolites of SalA in vivo is absent so far. In this study, by means of liquid chromatography with time-of-flight mass spectrometry (LC/TOFMS) and liquid chromatography with ion trap mass spectrometry (LC/MS(n)) techniques, the unknown metabolites of SalA in rat plasma after i.v. administration of the purified SalA at the dose of 20 mg/kg body weight were identified. A liquid-liquid extraction method was established to separate the metabolites from the plasma and the chromatographic separations were performed on a Xterra MS C(18) column (100 mm x 4.6 mm i.d., 3.5 microm) with acetonitrile/methanol/water/formic acid (20.5:19.5:64: 0.05, v/v/v/v) as the mobile phase at a constant flow rate of 0.2 mL/min. Based on the data obtained from the LC/TOFMS determination (the total ion chromatograms, MS spectra and extracted ion chromatograms), in combination with the characteristic fragment ions acquired from the LC/MS(n) determination, five metabolites were identified as SalA-monoglucuronide, monomethyl-SalA-monoglucuronide, mono-methyl-SalA, dimethyl-SalA and dimethyl-SalA-monoglucuronide, and the possible chemical structures were deduced. The results indicated that SalA might mainly undergo two metabolic pathways in vivo in rats, which were methylation and glucuronidation. The present studies have laid a solid foundation for the metabolic mechanism of SalA in vivo. PMID:19437443

Shen, Yi; Wang, Xueyan; Xu, Lihua; Liu, Xiaowei; Chao, Ruobing



Evaluation and application of static headspace-multicapillary column-gas chromatography-ion mobility spectrometry for complex sample analysis.  


An evaluation of static headspace-multicapillary column-gas chromatography-ion mobility spectrometry (SHS-MCC-GC-IMS) has been undertaken to assess its applicability for the determination of 32 volatile compounds (VCs). The key experimental variables of sample incubation time and temperature have been evaluated alongside the MCC-GC variables of column polarity, syringe temperature, injection temperature, injection volume, column temperature and carrier gas flow rate coupled with the IMS variables of temperature and drift gas flow rate. This evaluation resulted in six sets of experimental variables being required to separate the 32 VCs. The optimum experimental variables for SHS-MCC-GC-IMS, the retention time and drift time operating parameters were determined; to normalise the operating parameters, the relative drift time and normalised reduced ion mobility for each VC were determined. In addition, a full theoretical explanation is provided on the formation of the monomer, dimer and trimer of a VC. The optimum operating condition for each VC calibration data was obtained alongside limit of detection (LOD) and limit of quantitation (LOQ) values. Typical detection limits ranged from 0.1ng bis(methylthio)methane, ethylbutanoate and (E)-2-nonenal to 472ng isovaleric acid with correlation coefficient (R(2)) data ranging from 0.9793 (for the dimer of octanal) through to 0.9990 (for isobutyric acid). Finally, the developed protocols were applied to the analysis of malodour in sock samples. Initial work involved spiking an inert matrix and sock samples with appropriate concentrations of eight VCs. The average recovery from the inert matrix was 101±18% (n=8), while recoveries from the sock samples were lower, that is, 54±30% (n=8) for sock type 1 and 78±24% (n=6) for sock type 2. Finally, SHS-MCC-GC-IMS was applied to sock malodour in a field trial based on 11 volunteers (mixed gender) over a 3-week period. By applying the SHS-MCC-GC-IMS database, four VCs were identified and quantified: ammonia, dimethyl disulphide, dimethyl trisulphide and butyric acid. A link was identified between the presence of high ammonia and dimethyl disulphide concentrations and a high malodour odour grading, that is, ? 6. Statistical analysis did not find any correlation between the occurrence of dimethyl disulphide and participant gender. PMID:24630058

Denawaka, Chamila J; Fowlis, Ian A; Dean, John R



Characterization of fatty acid and triacylglycerol composition in animal fats using silver-ion and non-aqueous reversed-phase high-performance liquid chromatography/mass spectrometry and gas chromatography/flame ionization detection.  


Fatty acid (FA) and triacylglycerol (TG) composition of natural oils and fats intake in the diet has a strong influence on the human health and chronic diseases. In this work, non-aqueous reversed-phase (NARP) and silver-ion high-performance liquid chromatography with atmospheric pressure chemical ionization mass spectrometry detection and gas chromatography with flame-ionization detection (GC/FID) and mass spectrometry detection are used for the characterization of FA and TG composition in complex samples of animal fats from fallow deer, red deer, sheep, moufflon, wild boar, cock, duck and rabbit. The FA composition of samples is determined based on the GC/FID analysis of FA methyl esters. In total, 81 FAs of different acyl chain length, double bond (DB) number, branched/linear, cis-/trans- and DB positional isomers are identified. TGs in animal fats contain mainly monounsaturated and saturated FAs. High amounts of branched and trans-FAs are observed in the samples of ruminants. In NARP mode, individual TG species are separated including the separation of trans- and branched TGs. Silver-ion mode provides the separation of TG regioisomers, which enables the determination of their ratios. Great differences in the preference of unsaturated and saturated FAs in the sn-2 position on the glycerol skeleton are observed among individual animal fats. Unsaturated FAs are preferentially occupied in the sn-2 position in all animal samples except for wild boar with the strong preference of saturated FAs in the sn-2 position. PMID:21835413

Lísa, Miroslav; Netušilová, Kate?ina; Fran?k, Lukáš; Dvo?áková, Hana; Vrkoslav, Vladimír; Hol?apek, Michal



Determination of amino acids in the foods by reversed-phase high-performance liquid chromatography with a new precolumn derivative, butylthiocarbamyl amino acid, compared to the conventional phenylthiocarbamyl derivatives and ion-exchange chromatography.  


A new derivatizing reagent, butylisothiocyanate (BITC), reacts quantitatively with the 22 standard amino acids and the amino acids in the acid hydrolysate of food and the protein standard, bovine serum albumin (BSA), at 40 degrees C for 30 min to yield butylthiocarbamyl (BTC) amino acids. The sensitivity of BTC-amino acid was similar to phenylthiocarbamyl (PTC) amino acid. The detection limit in both derivatives was about 3.9 pmol at 0.05 AUFS that showed a stable baseline for the quantitative determination. Analysis of the results obtained with BSA and food samples as BTC derivatives showed good agreement with those determined as PTC derivatives, ion-exchange chromatography and data presented in the literatures, except for a few amino acids. Especially the values compared to those of ion-exchange chromatography were very close, except for histidine. The advantage of the BITC reagent over the phenylisothiocyanate was that it had high volatility, so the excess reagent and by-products were removed in about 10 min, compared to about 1 h in the PITC reagent, with a common aspirator. In the BTC derivatives, cystine and cysteine were determined separately but in the PTC derivatives they were resolved into a single peak. PMID:8765708

Woo, K L; Hwang, Q C; Kim, H S



Assay procedures for the determination of biogenic amines and their metabolites in rat hypothalamus using ion-pairing reversed-phase high-performance liquid chromatography.  


Procedures are described for the simultaneous determination, by high-performance liquid chromatography of adrenaline, noradrenaline, dopamine and 5-hydroxytryptamine using laurylsulphate as pairing ion and for the separate simultaneous determination of vanillylmandelic acid, dihydroxyphenylacetic acid, 3-methoxy-4-hydroxyphenylethylene glycol, 5-hydroxyindoleacetic acid and homovanillic acid using tetraethylammonium as pairing ion. Sample preparation consists of protein precipitation only and octadecylsilane coated silica is used for both sets of determinations as is electrochemical detection. The chromatographic basis of each separation is discussed in the light of modern ideas of ion exchange and desolvation to enable the method to be modified chromatographically if other compounds are to be resolved or the assay method modified. The quantitative aspects of the methods are detailed and applied to amine and metabolite levels in rat hypothalamus. The values determined together with their sample variation and estimated limits of detection are quoted. PMID:6315748

Taylor, R B; Reid, R; Kendle, K E; Geddes, C; Curle, P F



Simultaneous detection and quantitation of highly water-soluble herbicides in serum using ion-pair liquid chromatography-tandem mass spectrometry.  


We report the simultaneous screening of highly polar, water-soluble, and less-volatile herbicides, including glyphosate, glufosinate, paraquat, and diquat, in serum using liquid chromatography-mass spectrometry. The herbicides were separated by solid-phase extraction using a Strata-XC cartridge. A heptafluorobutyric acid solution was chosen as the mobile phase for ion-pair liquid chromatography. Mass spectrometry was used for analysis and was optimized for operation in the positive mode for all analytes. The serum specimens were screened for the presence of the herbicides at the following concentrations: 5 ng/mL for glyphosate, 2 ng/mL for glufosinate, 1 ng/mL for diquat, and 5 ng/mL for paraquat. This is the first report on the simultaneous detection of these compounds. PMID:19022710

Wang, Kuang-Chuan; Chen, Shih-Ming; Hsu, Jung-Fa; Cheng, Sheaw-Guey; Lee, Ching-Kuo



Determination of microbial fatty acid profiles at femtomolar levels in human urine and the initial marine microfouling community by capillary gas chromatography-chemical ionization mass spectrometry with negative ion detection  

Microsoft Academic Search

Summary Room temperature esterification with the electron capturing pentafluorobenzyl bromide in glass capillaries, with analysis by capillary gas-liquid chromatography coupled with chemical ionization mass spectrometry and negative ion detection in the selected ion mode, allowed detection and identification of fatty acids from micro- bial biofilms at the femtomolar level. This sensitivity was achieved without loss of specificity of the mass

Goran Odham; Anders Tunlid; Gunilla Westerdahl; Lennart Larsson; James B. Guckert; David C. White



Simultaneous ionization and analysis of 84 anabolic androgenic steroids in human urine using liquid chromatography-silver ion coordination ionspray/triple-quadrupole mass spectrometry.  


Metal ion coordination ionspray (M(+) CIS) ionization is a powerful technique to enhance ionization efficiency and sensitivity. In this study, we developed and validated an analytical method for simultaneous ionization and analysis of 84 anabolic androgenic steroids (65 exogenous and 19 endogenous) using liquid chromatography-silver ion coordination ionspray/triple-quadrupole mass spectrometry (LC-Ag(+) CIS/MS/MS). The concentrations of silver ions and organic solvents have been optimized to increase the amount of silver ion coordinated complexes. A combination of 25??M of silver ions and methanol showed the best sensitivity. The validation results showed the intra- (0.8-9.2%) and inter-day (2.5-14.9%) precisions, limits of detection (0.0005-5.0?ng/mL), and matrix effect (71.8-100.3%) for the screening analysis. No significant ion suppression was observed. In addition, this method was successfully applied to analysis of positive samples from suspected abusers and useful for the detection of the trace levels of anabolic steroids in human urine samples. Copyright © 2014 John Wiley & Sons, Ltd. PMID:25382000

Kim, So-Hee; Cha, Eun-Ju; Lee, Kang Mi; Kim, Ho Jun; Kwon, Oh-Seung; Lee, Jaeick



Reversed-phase high-performance liquid chromatography of catecholamines and their congeners with simple acids as ion-pairing reagents.  


We have investigated the value of various common acids as ion-pairing reagents for high-efficiency separations of catecholamines and their metabolites in reversed-phase high-performance liquid chromatography. The retention of norepinephrine, alpha-methylnorepinephrine, dopamine, alpha-methyldopamine, L-dopa, alpha-methyldopa, dihydroxybenzylamine, epinephrine, carbidopa and DOPAC was measured in mobile phases composed of nitric, sulfuric, acetic and trichloroacetic acids at pH 2-5 and anion concentrations ranging from 5-500 mM. The solute capacity ratios were dependent on the hydrophobicity and concentration of the ion-pairing reagent and the pH of the mobile phase. Good retention, peak symmetry and high efficiency (3000 theoretical plates for 300 mm) was found for mobile phases composed of the strong inorganic acids and trichloroacetic acid. Chromatography was compared to that seen using the detergent sodium octylsulfate. Trichloroacetic acid gave retention and efficiency similar to sodium octylsulfate. These experiments show that simple acids can replace alkylsulfates as ion-pairing reagents for the separation of the catecholamines and their metabolites. PMID:43866

Asmus, P A; Freed, C R



Field detection of bacillus spore aerosols with stand-alone pyrolysis-gas chromatography and ion mobility spectrometry  

NASA Astrophysics Data System (ADS)

A commercially available, hand-held chemical vapor detector was modified to detect Gram-positive Bacillus subtilis var. globigii spores (BG) in outdoor field scenarios. An Airborne Vapor Monitor (AVM) ion mobility spectrometry (IMS) vapor detector was interfaced to a biological sample processing and transfer introduction system. The biological sample processing was accomplished by quartz tube pyrolysis (Py), and the resultant vapor was transferred by gas chromatography (GC) to the IMS detector. The Py-GC/IMS system can be described as a hyphenated device where two analytical dimensions, in series, allow the separation and isolation of individual components from the pyrolytic decomposition of biological analytes. Gram positive spores such as BG contain 5 - 15% by weight of dipicolinic acid (DPA), and picolinic acid is a pyrolysis product of DPA. Picolinic acid has a high proton affinity, and it is detected in a sensitive fashion by the atmospheric pressure-based IMS device. Picolinic acid occupies a unique region in the GC/IMS data domain with respect to other bacterial pyrolysis products. A 1000 to 1, air-to-air, aerosol concentrator was interfaced to the Py-GC/IMS instrument, and the system was placed in an open-air, Western United States desert environment. The system was tested with BG spore aerosol releases, and the instrument was remotely operated during a trial. A Met-One aerosol particle counter was placed next to the Py-GC/IMS so as to obtain a real-time record of the ambient and bacterial aerosol challenges. The presence/absence of an aerosol event, determined by an aerosol particle counter and a slit sampler-agar plate system, was compared to the presence/absence of a picolinic acid response in a GC/IMS data window at selected times in a trial with respect to a BG challenge. In the 21 BG trials, the Py-GC/IMS instrument experienced two true negatives, no false positives, and the instrument developed a software failure in one trial. The remaining 18 trials were true positive determinations for the presence of BG aerosol, and a limit of detection for the Py-GC/IMS instrument was estimated at approximately 3300 BG spore-containing particles.

Snyder, A.; Maswadeh, Waleed M.; Parsons, John A.; Tripathi, Ashish; Meuzelaar, Henk L. C.; Dworzanski, Jacek P.; Kim, Man-Goo



Comparison of a pulsed electron capture detector and a Fourier transform ion mobility detector after capillary supercritical fluid chromatography  

Microsoft Academic Search

A commercial electron capture detector and an ion mobility detector were interfaced to a supercritical fluid chromatograph (SFC). The ion mobility detector (IMD) was operated in the negative ion collection mode wherein analyte molecules of high electron affinity form negatively charged ions in the detector, producing an ECD-like response. Responses of the two detectors were compared using pressure programmed SFC

E. E. Tarver; H. H. Hill



Journal of Chromatography A, 1043 (2004) 249254 Arsenic complexes in the arsenic hyperaccumulator  

E-print Network

Journal of Chromatography A, 1043 (2004) 249­254 Arsenic complexes in the arsenic hyperaccumulator-exchange chromatography­hydride generation­atomic fluorescence spectroscopy and size-exclusion chromatographyIII-tris-thiolate complexes through thiolate bonds by us- ing size-exclusion chromatography (SEC) or electrospray ionization

Ma, Lena


Negative ion chemical ionization gas chromatography/mass spectrometry and gas chromatography/tandem mass spectrometry of prostanoid pentafluorobenzyl ester/methoxime/trimethylsilyl ether derivatives.  


Negative ion chemical ionization mass spectra of prostaglandin F2 alpha, prostaglandin E2, prostaglandin D2, 6-oxo-prostaglandin F1 alpha, 2,3-dinor-6-oxo-prostaglandin F1 alpha, thromboxane B2, 2,3-dinor-thromboxane B2, and 11-dehydro-thromboxane B2 pentafluorobenzyl ester (PFB)/methoxime/trimethylsilyl ether derivatives are presented. Collisionally activated decomposition mass spectra of the [M-PFB]- ions at collision energies of 8-24 eV and argon collision gas pressures of 1-2m Torr almost show only fragmentation of trimethylsilanol, (CH3)2Si = CHOH, (CH3)2Si = CH2 and methanol whereas, except for carbon dioxide loss, only few low-intensity fragments from the carbon skeleton of the prostanoids are observed. PMID:3365491

Schweer, H; Seyberth, H W; Meese, C O; Fürst, O



Determination of trace labile copper in environmental waters by magnetic nanoparticle solid phase extraction and high-performance chelation ion chromatography.  


Cobalt magnetic nanoparticles surface functionalised with iminodiacetic acid were evaluated as a nano-particulate solid phase extraction absorbent for copper ions (Cu(2+)) from environmental water samples. Using an external magnetic field, the collector nanoparticles could be separated from the aqueous phase, and adsorbed ions simply decomplexed using dilute HNO3. Effects of pH, buffer concentration, sample and sorbent volume, extraction equilibrium time, and interfering ion concentration on extraction efficiency were investigated. Optimal conditions were then applied to the extraction of Cu(2+) ions from natural water samples, prior to their quantitation using high-performance chelation ion chromatography. The limits of detection (LOD) of the combined extraction and chromatographic method were ~0.1ngml(-1), based upon a 100-fold preconcentration factor (chromatographic performance; LOD=9.2ngml(-1) Cu(2+)), analytical linear range from 20 to 5000ngmL(-1), and relative standard deviations=4.9% (c=1000ngml(-1), n=7). Accuracy and precision of the combined approach was verified using a certified reference standard estuarine water sample (SLEW-2) and comparison of sample determinations with sector field inductively coupled plasma mass spectrometry. Recoveries from the addition of Cu(2+) to impacted estuarine and rain water samples were 103.5% and 108.5%, respectively. Coastal seawater samples, both with and without prior UV irradiation and dissolved organic matter removal were also investigated using the new methodology. The effect of DOM concentration on copper availability was demonstrated. PMID:25640140

Wei, Z; Sandron, S; Townsend, A T; Nesterenko, P N; Paull, B



Chelation ion chromatography of alkaline earth and transition metals a using monolithic silica column with bonded N-hydroxyethyliminodiacetic acid functional groups.  


A commercially available porous silica monolithic column (Onyx Monolithic Si, 100 mm×4.6 mm I.D.) was 'in-column' covalently functionalised with 2-hydroxyethyliminodiacetic acid (HEIDA) groups, and applied to the simultaneous and rapid separation of alkaline earth and transition metal ions, using high-performance chelation ion chromatography (HPCIC). With a 0.3mM dipicolinic acid (DPA) containing eluent, the baseline separation of various common transition and heavy metal ions and the four alkaline earth metal ions could be achieved in under 14 min with a flow rate of just 0.8 mL/min. Detection was achieved using spectrophotometric detection at 540 nm after post-column reaction (PCR) with 4-(2-pyridylazo)-resorcinol (PAR). Significant effects from variation of eluent nature, concentration and temperature upon selectivity and retention were demonstrated with the new monolithic silica chelating phase. Under optimised conditions (0.165 M LiNO(3) eluent, pH 2.5), peak efficiencies of 54,000, 60,000 and 64,000 N/m, for Zn(2+), Mn(2+) and Cd(2+), respectively, were recorded, far exceeding that previously reported for IDA based chelation ion exchange columns. PMID:23298846

McGillicuddy, Nicola; Nesterenko, Ekaterina P; Nesterenko, Pavel N; Jones, Phil; Paull, Brett



Separation and detection of compounds in Honeysuckle by integration of ion-exchange chromatography fractionation with reversed-phase liquid chromatography-atmospheric pressure chemical ionization mass spectrometer and matrix-assisted laser desorption\\/ionization time-of-flight mass spectrometry analysis  

Microsoft Academic Search

A hyphenated method for the isolation and identification of components in a traditional Chinese medicine of Honeysuckle was developed. Ion-exchange chromatography (IEC) was chosen for the fractionation of Honeysuckle extract, and then followed by concentration of all the fractions with rotary vacuum evaporator. Each of the enriched fractions was then further analyzed by reversed-phase liquid chromatography-atmospheric pressure chemical ionization mass

Xueguo Chen; Lianghai Hu; Xingye Su; Liang Kong; Mingliang Ye; Hanfa Zou



Discrimination of crude and processed rhubarb products using a chemometric approach based on ultra fast liquid chromatography with ion trap/time-of-flight mass spectrometry.  


Crude rhubarb subjected to different processing procedures will produce different therapeutic effects that are possibly due to processing-induced variation in chemical composition. In this study, a chemometric approach based on ultra fast liquid chromatography with ion trap/time-of-flight mass spectrometry was established to systematically investigate the chemical variations of rhubarb induced by different processing methods. The approach was validated based on pooled quality-control samples from two perspectives: the individual properties of variables and the bulk properties of samples. Orthogonal partial least squares discriminant analysis was introduced to compare the differences between crude and processed rhubarb products. A total of 20 significantly different markers were screened out and unambiguously/tentatively characterized. This research proved that a chemometric method based on ultra fast liquid chromatography with ion trap/time-of-flight mass spectrometry can comprehensively analyze the chemical variation of herbal medicine and provide evidence for a deeper understanding of the pharmacological activities of processed rhubarb products. PMID:25421806

Wang, Min; Fu, Jinfeng; Guo, Huimin; Tian, Yuan; Xu, Fengguo; Song, Rui; Zhang, Zunjian



The identification and quantification of a high molecular weight light stabilizer in polycarbonate by application of an online coupling of size exclusion chromatography in stopped flow mode with pyrolysis gas chromatography time of flight mass spectroscopy.  


The identification and quantification of a high molecular weight light stabilizer (Uvinul 3030™) in an unknown polycarbonate sample was achieved through the application of SEC-Py-TOF-GCMS. A size exclusion column optimized to achieve resolution in the lower mass range was applied to allow the fractionation of an individual additive peak. A commercially available sampling interface was operated in stop flow mode and fractions were pyrolyzed to allow chromatographic separation of the fragments of the otherwise non-volatile stabilizer. After identification on the basis of accurate mass and elemental composition of the additive the quantification was compared using the available SEC-UV and SEC-PY-GC-TOFMS data. The resulting method provided a high degree of certainty in identification and flexibility in quantification expected to be applicable to other additives of similar volatilities or functional class. PMID:25160954

Brander, Eric; Wold, Christian



Determination of site selectivity of different carcinogens for preferential mutational hot spots in oligonucleotide fragments by ion-pair reversed-phase nano liquid chromatography tandem mass spectrometry.  


Ion-pair reversed-phase nano liquid chromatography coupled with nanospray ion trap mass spectrometry was used to investigate site selectivity of the known carcinogens N-acetoxy-2-acetylaminofluorene, N-hydroxy-4-aminobiphenyl and (+/-)-anti-benzo[a]pyrene diol epoxide with the synthetic double-strand 14-mer long oligonucleotide fragment of the p53 gene containing two mutational hot-spot codons (5'-P-ACC155 CGC156 GTC157 CGC158 GC/5'-GCG CGG ACG CGG GT). The investigation was performed using a monolithic polystyrene divinylbenzene capillary column and triethylammonium bicarbonate as an ion-pair reagent. The exact location of the carcinogen on the modified oligonucleotide backbone was determined using characteristic collision-induced dissociation fragmentation patterns obtained under negative-ion mode ionization. In all these cases, the adducted, isomeric oligonucleotides formed were chromatographically resolved and structural identification was performed without any prior deoxyribonucleic acid cleavage or hydrolysis. The knowledge of the site specificity of a carcinogen, especially at purported mutational hot spots, is of paramount importance (1) in establishing the identity of biomarkers for an early risk assessment of the formed DNA adducts, (2) developing repair mechanisms for the formed carcinogen adducted DNA, and (3) understanding the nature of the covalent bond formed and mapping the frequency of the adduction process. PMID:24881456

Sharma, Vaneet K; Xiong, Wennan; Glick, James; Vouros, Paul



Chromatography Nomenclature and Definitions  

NSDL National Science Digital Library

This website gives the International Union of Pure and Applied Chemistry approved definitions in the field of chromatography. It is critical for students to appreciate the importance of using standardized nomenclature and definitions. Sections of the site include general terminology, terms related to the chromatographic system, terms related to the chromatographic process and the theory of chromatography, terms related to detection, ion exchange, liquid-liquid distribution (solvent extraction) and other related subjects.


Effect of humic acid on absorption-release processes in the bottom sediments-Yenisei river water system as studied by dual-column ion chromatography and ?-ray spectrometry  

Microsoft Academic Search

The effect of humic acid on absorption-release processes in the bottom sediments-Yenisei river water system was studied by\\u000a dual-column ion chromatography and ?-ray spectrometry. With the use of ion chromatography, it was found that processes related\\u000a to the absorption of SO\\u000a 4\\u000a 2?\\u000a and Cl? anions by a solid phase with the release of NO\\u000a 3\\u000a ?\\u000a , PO\\u000a 4

L. G. Bondareva; O. P. Kalyakina; A. Ya. Bolsunovskii



Applying Chromatography.  

ERIC Educational Resources Information Center

Presents three experiments to introduce students to different kinds of chromatography: (1) paper chromatography; (2) gel filtration chromatography; and (3) reverse-phase liquid chromatography. Written in the form of a laboratory manual, explanations of each of the techniques, materials needed, procedures, and a glossary are included. (PVD)

Klein, Jessie W.; Patev, Paul



ORIGINAL PAPER Development of a methodology utilizing gas chromatography  

E-print Network

ORIGINAL PAPER Development of a methodology utilizing gas chromatography ion-trap tandem mass chromatography coupled with an ion-trap tandem mass spectrometry detection system (IT-MS/MS) was developed. Four

Mazumder, Asit


Target-guided separation of Bougainvillea glabra betacyanins by direct coupling of preparative ion-pair high-speed countercurrent chromatography and electrospray ionization mass-spectrometry.  


In this study, preparative ion-pair high-speed countercurrent chromatography was directly coupled to an electrospray ionization mass-spectrometry device (IP-HSCCC/ESI-MS-MS) for target-guided fractionation of high molecular weight acyl-oligosaccharide linked betacyanins from purple bracts of Bougainvillea glabra (Nyctaginaceae). The direct identification of six principal acyl-oligosaccharide linked betacyanins in the mass range between m/z 859 and m/z 1359 was achieved by positive ESI-MS ionization and gave access to the genuine pigment profile already during the proceeding of the preparative separation. Inclusively, all MS/MS-fragmentation data were provided during the chromatographic run for a complete analysis of substitution pattern. On-line purity evaluation of the recovered fractions is of high value in target-guided screening procedures and for immediate decisions about suitable fractions used for further structural analysis. The applied preparative hyphenation was shown to be a versatile screening method for on-line monitoring of countercurrent chromatographic separations of polar crude pigment extracts and also traced some minor concentrated compounds. For the separation of 760mg crude pigment extract the biphasic solvent system tert.-butylmethylether/n-butanol/acetonitrile/water 2:2:1:5 (v/v/v/v) was used with addition of ion-pair forming reagent trifluoroacetic acid. The preparative HSCCC-eluate had to be modified by post-column addition of a make-up solvent stream containing formic acid to reduce ion-suppression caused by trifluoroacetic acid and later significantly maximized response of ESI-MS/MS detection of target substances. A variable low-pressure split-unit guided a micro-eluate to the ESI-MS-interface for sensitive and direct on-line detection, and the major volume of the effluent stream was directed to the fraction collector for preparative sample recovery. The applied make-up solvent mixture significantly improved smoothness of the continuously measured IP-HSCCC-ESI-MS base peak ion trace in the experimental range of m/z 50-2200 by masking stationary phase bleeding and generating a stable single solvent phase for ESI-MS/MS detection. Immediate structural data were retrieved throughout the countercurrent chromatography run containing complete MS/MS-fragmentation pattern of the separated acyl-substituted betanidin oligoglycosides. Single ion monitoring indicated clearly the base-line separation of higher concentrated acylated betacyanin components. PMID:20494364

Jerz, Gerold; Wybraniec, S?awomir; Gebers, Nadine; Winterhalter, Peter



Applying 'Sequential Windowed Acquisition of All Theoretical Fragment Ion Mass Spectra' (SWATH) for systematic toxicological analysis with liquid chromatography-high-resolution tandem mass spectrometry.  


Liquid chromatography-tandem mass spectrometry (LC-MS/MS) has become an indispensable analytical technique in clinical and forensic toxicology for detection and identification of potentially toxic or harmful compounds. Particularly, non-target LC-MS/MS assays enable extensive and universal screening requested in systematic toxicological analysis. An integral part of the identification process is the generation of information-rich product ion spectra which can be searched against libraries of reference mass spectra. Usually, 'data-dependent acquisition' (DDA) strategies are applied for automated data acquisition. In this study, the 'data-independent acquisition' (DIA) method 'Sequential Windowed Acquisition of All Theoretical Fragment Ion Mass Spectra' (SWATH) was combined with LC-MS/MS on a quadrupole-quadrupole-time-of-flight (QqTOF) instrument for acquiring informative high-resolution tandem mass spectra. SWATH performs data-independent fragmentation of all precursor ions entering the mass spectrometer in 21m/z isolation windows. The whole m/z range of interest is covered by continuous stepping of the isolation window. This allows numerous repeat analyses of each window during the elution of a single chromatographic peak and results in a complete fragment ion map of the sample. Compounds and samples typically encountered in forensic casework were used to assess performance characteristics of LC-MS/MS with SWATH. Our experiments clearly revealed that SWATH is a sensitive and specific identification technique. SWATH is capable of identifying more compounds at lower concentration levels than DDA does. The dynamic range of SWATH was estimated to be three orders of magnitude. Furthermore, the >600,000 SWATH spectra matched led to only 408 incorrect calls (false positive rate?=?0.06 %). Deconvolution of generated ion maps was found to be essential for unravelling the full identification power of LC-MS/MS with SWATH. With the available software, however, only semi-automated deconvolution was enabled, which rendered data interpretation a laborious and time-consuming process. PMID:25366975

Arnhard, Kathrin; Gottschall, Anna; Pitterl, Florian; Oberacher, Herbert



Determination of free desmosine and isodesmosine as urinary biomarkers of lung disorder using ultra performance liquid chromatography-ion mobility-mass spectrometry.  


The elastin degradation products, desmosine (DES) and isodesmosine (IDES) are highly stable, cross-linking amino-acids that are unique to mature elastin. The excretion of DES/IDES in urine, in the free form and with associated peptide fragments, provides an indicator of lung damage in chronic obstructive pulmonary disease (COPD). A quantitative ion mobility-mass spectrometry (IM-MS) method has been developed for the analysis of free DES/IDES in urine with deuterated IDES as an internal standard. Resolution of DES/IDES isomers was achieved in less than five minutes using ultra performance liquid chromatography (UPLC) combined with ion pairing. The optimized UPLC-IM-MS method provided a linear dynamic range of 10-300 ng/mL and a limit of quantitation of 0.028 ng/mL for IDES and 0.03 ng/mL for DES (0.55 ng and 0.61 ng on column respectively). The method reproducibility (%RSD) was <4% for DES and IDES. The UPLC-IM-MS method was applied to the analysis of urine samples obtained from healthy volunteers and COPD patients. The DES/IDES concentrations in healthy and COPD urine showed an increase in DES (79%) and IDES (74%) in the COPD samples, relative to healthy controls. The incorporation of an IM separation prior to m/z measurement by MS was shown to reduce non-target ion responses from the bio-fluid matrix. PMID:22088351

Devenport, Neil A; Reynolds, James C; Parkash, Ved; Cook, Jason; Weston, Daniel J; Creaser, Colin S



[Screening and confirmation of 24 hormones in cosmetics by ultra high performance liquid chromatography-linear ion trap/orbitrap high resolution mass spectrometry].  


A method of ultra high performance liquid chromatography-linear ion trap/orbitrap high resolution mass spectrometry (UPLC-LTQ/Orbitrap MS) was established to screen and confirm 24 hormones in cosmetics. Various cosmetic samples were extracted with methanol. The extract was loaded onto a Waters ACQUITY UPLC BEH C18 column (50 mm x 2.1 mm, 1.7 microm) using a gradient elution of acetonitrile/water containing 0.1% (v/v) formic acid for the separation. The accurate mass of quasi-molecular ion was acquired by full scanning of electrostatic field orbitrap. The rapid screening was carried out by the accurate mass of quasi-molecular ion. The confirmation analysis for targeted compounds was performed with the retention time and qualitative fragments obtained by data dependent scan mode. Under the optimal conditions, the 24 hormones were routinely detected with mass accuracy error below 3 x 10(-6) (3 ppm), and good linearities were obtained in their respective linear ranges with correlation coefficients higher than 0.99. The LODs (S/N = 3) of the 24 compounds were < or = 10 microg/kg, which can meet the requirements for the actual screening of cosmetic samples. The developed method was applied to screen the hormones in 50 cosmetic samples. The results demonstrate that the method is a useful tool for the rapid screening and identification of the hormones in cosmetics. PMID:25185307

Li, Zhaoyong; Wang, Fengmei; Niu, Zengyuan; Luo, Xin; Zhang, Gang; Chen, Junhui



Detection and Chemical Profiling of Ling-Gui-Zhu-Gan Decoction by Ultra Performance Liquid Chromatography-Hybrid Linear Ion Trap-Orbitrap Mass Spectrometry.  


Ling-Gui-Zhu-Gan decoction (LGZGD), a well-known traditional Chinese medicine (TCM) formula, has been extensively used for the treatment of cardiovascular disease in clinic. However, the chemical constituents in LGZGD had not been investigated so far. In this study, an ultra performance liquid chromatography-hybrid electrospray ionization linear ion trap-Orbitrap mass spectrometry (UPLC-LTQ-Oribitrap-MS/MS) method was established for rapid separation and structural identification of the constituents in LGZGD. Separation was performed on an ACQUITY(TM) UPLC BEH C18 column (50 × 2.1 mm, 1.7 ?m) by gradient elution mode, using acetonitrile-water containing 0.1% formic acid as mobile phase at the flow rate of 0.2 mL/min. Accurate mass measurement for molecular ions and characteristic fragment ions could represent identification criteria for these compounds. As a result, 95 compounds including triterpene acids, triterpene saponins, flavonoids, coumarins, coumestans, benzofurans, phenylpropanoids and sesquiterpenoid lactones were detected, and 90 of them were tentatively identified. All compounds were further assigned in the individual raw material. In conclusion, the UPLC-LTQ-Orbitrap-MS/MS is a highly efficient technique to separate and identify constituents in complex matrices of TCMs. These results obtained in this research will provide a basis for quality control and further in vivo study of LGZGD. PMID:24920655

Wang, Pei; Wang, Bo; Xu, Jingyao; Sun, Jianbo; Yan, Qin; Ji, Bin; Zhao, Yunli; Yu, Zhiguo



Fulvic acid: modifier of metal-ion chemistry  

SciTech Connect

Fulvic acid, which is derived from the decay of plants and animals, is being studied for its role in the transport and toxicity of metal ions in soil and water. It is discussed in relation to the origin of humic substances and its interactions with metal ions. Techniques for investigating complexes of fulvic acid and metal ions are presented. They are separation and nonseparation analyses which are applied to speciation problems. The applicability, advantages, and disadvantages of both methods are presented. Separation of free and complexed metal ions can be done by chromatography, or with membranes that exclude the metal-ion complexes. Chromatographic techniques include liquid chromatography by size exclusion. Nonseparation techniques include voltametry and potentiometry, as well as fluorescence. A comparison of methods for calculation of the conditional stability constant K for complexes containing fulvic acid and copper (II) or cadmium (II) is presented.

Saar, R.A.; Weber, J.H.



Liquid chromatography/quadrupole ion trap/time-of-flight determination of the efficacy of drug test kits for rapid screening of food.  


The goal of this study was to evaluate the plausibility and accuracy of commercially available on-site immunoassay urinalysis kits for the screening of compounds of interest within food matrices. In conjunction with this study, a sensitive, robust, and reproducible analytical method, utilizing solid-phase extraction liquid chromatography/quadrupole ion trap/time-of-flight mass spectrometry for confirmation analysis, was developed. The food matrices analyzed were tomato juice, apple juice, milk, beer, white wine, ground beef, powdered milk, and all-purpose flour. Compounds fortified into the food matrices included heroin, phencyclidine, cocaine, benzoylecgonine, methadone, 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine, imipramine, doxepin, nitrazepam, diazepam, oxazepam, temazepam, alprazolam, flunitrazepam, clonazepam, and lorazepam. Standard curves were prepared for each matrix from 10 to 500 ng/ml for each analyzed compound. All liquid chromatography/tandem mass spectrometry samples were fortified with 20 microl of deuterated internal standard at 90 ng/ml. Quality control standards were prepared at 20 and 400 ng/ml, and > 90% were within 2 SD of the mean for each analyte. The test kits were found to produce up to 85% of the expected results based on concentration levels of adulterants (i-Screen in milk). This study shows that lateral-flow immunoassay test kits are plausible as a rapid, accurate, and reliable screening method in the event of adulteration of the food supply. PMID:18522037

Zuckschwerdt, Jessica B; Nixon, Christopher E; Ciner, Frederic L; Croley, Timothy R



Integration of ion-exchange chromatography fractionation with reversed-phase liquid chromatography-atmospheric pressure chemical ionization mass spectrometer and matrix-assisted laser desorption\\/ionization time-of-flight mass spectrometry for isolation and identification of compounds in Psoralea corylifolia  

Microsoft Academic Search

An approach for the separation and identification of components in a traditional Chinese medicine Psoralea corylifolia was developed. Ion-exchange chromatography (IEC) was applied for the fractionation of P. corylifolia extract, and then followed by concentration of all the fractions with rotary vacuum evaporator. Each of the enriched fractions was then further separated on an ODS column with detection of UV

Xueguo Chen; Liang Kong; Xingye Su; Chensong Pan; Mingliang Ye; Hanfa Zou



Quantitative protein analysis using (13)C7-labeled iodoacetanilide and d5-labeled N-ethylmaleimide by nano liquid chromatography/nanoelectrospray ionization ion trap mass spectrometry.  


We have developed a methodology for quantitative analysis and concurrent identification of proteins by the modification of cysteine residues with a combination of iodoacetanilide (IAA, 1) and (13)C7-labeled iodoacetanilide ((13)C7-IAA, 2), or N-ethylmaleimide (NEM, 3) and d5-labeled N-ethylmaleimide (d5-NEM, 4), followed by mass spectrometric analysis using nano liquid chromatography/nanoelectrospray ionization ion trap mass spectrometry (nano LC/nano-ESI-IT-MS). The combinations of these stable isotope-labeled and unlabeled modifiers coupled with LC separation and ESI mass spectrometric analysis allow accurate quantitative analysis and identification of proteins, and therefore are expected to be a useful tool for proteomics research. PMID:23562245

Kurono, Sadamu; Kaneko, Yuka; Niwayama, Satomi



Direct chemical-analysis of uv laser-ablation products of organic polymers by using selective ion monitoring mode in gas-chromatography mass-spectrometry  

USGS Publications Warehouse

Trace quantities of laser ablated organic polymers were analyzed by using commercial capillary column gas chromatography/mass spectrometry; the instrument was modified so that the laser ablation products could be introduced into the capillary column directly and the constituents of each peak in the chromatogram were identified by using a mass spectrometer. The present study takes advantage of the selective ion monitoring mode for significantly improving the sensitivity of the mass spectrometer as a detector, which is critical in analyzing the trace quantities and confirming the presence or absence of the species of interest in laser ablated polymers. The initial composition of the laser ablated polymers was obtained by using an electron impact reflectron time-of-flight mass spectrometer and the possible structure of the fragments observed in the spectra was proposed based on the structure of the polymers.

Choi, Y.; Lee, H.W.; Fountain, S.T.; Lubman, D.M.



Development and validation of automatic HS-SPME with a gas chromatography-ion trap/mass spectrometry method for analysis of volatiles in wines.  


An automated headspace solid-phase microextraction (HS-SPME) combined with gas chromatography-ion trap/mass spectrometry (GC-IT/MS) was developed in order to quantify a large number of volatile compounds in wines such as alcohols, ester, norisoprenoids and terpenes. The procedures were optimized for SPME fiber selection, pre-incubation temperature and time, extraction temperature and time, and salt addition. A central composite experimental design was used in the optimization of the extraction conditions. The volatile compounds showed optimal extraction using a DVB/CAR/PDMS fiber, incubation of 5 ml of wine with 2g NaCl at 45 °C during 5 min, and subsequent extraction of 30 min at the same temperature. The method allowed the identification of 64 volatile compounds. Afterwards, the method was validated successfully for the most significant compounds and was applied to study the volatile composition of different white wines. PMID:23158309

Paula Barros, Elisabete; Moreira, Nathalie; Elias Pereira, Giuliano; Leite, Selma Gomes Ferreira; Moraes Rezende, Claudia; Guedes de Pinho, Paula



Simultaneous quantification of sinigrin, sinalbin, and anionic glucosinolate hydrolysis products in Brassica juncea and Sinapis alba seed extracts using ion chromatography.  


Although mustards such as Sinapis alba and Brassica juncea contain glucosinolates (sinalbin and sinigrin, respectively) that hydrolyze to form biopesticidal products, routine quality control methods to measure active ingredients in seed and seed meals are lacking. We present a simple and fast ion chromatography method for the simultaneous quantification of sinigrin, sinalbin, and anionic hydrolysis products in mustard seed to assess biological potency. Optimum conditions include isocratic elution with 100 mM NaOH at a flow rate of 0.9 mL/min on a 4 × 210 mm hydroxide-selective anion-exchange column. All anion analytes including sinigrin, sinalbin, SO4(2-), and SCN(-) yielded recoveries ranging from 83 to 102% and limits of detection ?0.04 mM, with samples displaying little interference from plant matrix components. Sample preparation is minimized and analysis times are shortened to <90 min as compared with previous methods that took days and multiple instruments. PMID:25314611

Popova, Inna E; Morra, Matthew J



The determination of potassium concentration in vitreous humor by low pressure ion chromatography and its application in the estimation of postmortem interval.  


An analytical method was developed for the determination of potassium in vitreous humor by low pressure ion chromatography (LPIC). Experimental conditions for LPIC analysis were optimized. High sensitivity and selectivity were obtained using this method. The LOD and LOQ were 1 and 2 mmol l(-1), respectively. The linearity was demonstrated from 2 to 20 mmol l(-1). The intra- and inter-day precision (CV) based on three concentrations was less than 5.0%. It was a simple and fast method to measure potassium and was suitable for evaluating the postmortem interval (PMI) in relatively well-preserved bodies. Sixty-two samples from medical-legal autopsies with known PMI were analyzed. A linear correlation equation for potassium concentration in the vitreous humor and PMI was established: [K(+)]=0.1702PMI+5.5678, r=0.8692. PMID:17347058

Zhou, Bin; Zhang, Lin; Zhang, Gengqian; Zhang, Xinshen; Jiang, Xiaoping



Analysis of earthy and musty odors in water samples by solid-phase microextraction coupled with gas chromatography/ion trap mass spectrometry.  


A method for the determination of the earthy and musty odors geosmin, 2-methylisoborneol (2-MIB), 2-isobutyl-3-methoxy pyrazine (IBMP), 2-isopropyl-3-methoxy pyrazine (IPMP) and 2,4,6-trichloroanisole (2,4,6-TCA) in water by headspace solid-phase microextraction (HSSPME) combined with gas chromatography-ion trap mass spectrometry (GC-ITMS) is described. Several parameters of the extraction and desorption procedure were studied and optimized (such as types of fibers, extraction temperature, extraction time, desorption temperature, desorption time, ionic strength and elutropic strength and pH of samples). The method shows good linearity over the concentration range 1-500ngl(-1) and gives detection limits of sub-part per trillion levels for all compounds. Good precision (5.9-9.8%) is obtained using IBMP as internal standard. Finally, the method was successfully applied to analyze earthy and musty odors in tap water and lake water. PMID:18969829

Sung, Yu-Hsiang; Li, Tzu-Ying; Huang, Shang-Da



Targeted profiling of polar intracellular metabolites using ion-pair-high performance liquid chromatography and -ultra high performance liquid chromatography coupled to tandem mass spectrometry: applications to serum, urine and tissue extracts.  


The effective analysis of polar ionic metabolites by LC-MS, such as those encountered in central carbon metabolism, represents a major problem for metabolic profiling that is not adequately addressed using strategies based on either reversed-phase or HILIC methods. Here we have compared analysis of central carbon metabolites on optimized methods using HILIC, porous graphitic carbon or ion pair chromatography (IPC) using tributyl ammonium as IP reagent. Of the 3 chromatographic approaches examined only IPC enabled us to obtain a robust analytical methodology. This system was used to profile more than a hundred endogenous metabolic intermediates in urine, serum and tissue samples. However, whilst we found IPC to be the best of the approaches examined considerable care was still needed to obtain robust data. Thus, in excess of 40 of representative biological samples were needed to "condition" a new analytical column and further 10 matrix injections were then required at the beginning of each analytical batch in order to obtain robust and reproducible chromatographic separations. An additional limitation that we have found was that, for a small number of phosphorylated and poly carboxylic acid metabolites, measurement was only possible if the analytes were present in relatively high concentrations. We also found that, whilst this methodology could be used for the analysis of both in vitro cell culture media, cell extracts, tissue, and biological fluids (blood, urine), for the best results columns should only be used to analyze a single matrix. However, despite the need for extensive column conditioning, and the manifold disadvantages resulting from the contamination of the separation system and mass spectrometer with the ion pair reagent, IPC-MS currently provides the best means of analyzing these polar, ionic and problematic metabolites. PMID:24861786

Michopoulos, Filippos; Whalley, Nicky; Theodoridis, Georgios; Wilson, Ian D; Dunkley, Tom P J; Critchlow, Susan E



Metabolism of cymoxanil and analogs in strains of the fungus Botrytis cinerea using high-performance liquid chromatography and ion-pair high-performance thin-layer chromatography.  


The metabolism of cyano-oxime fungicide 1-(2-cyano-2-methoxyiminoacetyl)-3-ethylurea (cymoxanil) and analogs was studied on several strains of the fungus Botrytis cinerea owing to their difference in sensitivity towards cymoxanil. Chromatographic analysis of the unextracted culture medium was simpler and more accurate, particularly for ionizable metabolites because it avoids problems associated with extraction. Reversed-phase high-performance liquid chromatography was applied to compare the decrease of cymoxanil and analogs caused by different strains of B. cinerea, by periodic injections of incubated culture medium aliquots, directly on a C4 wide-pore column. Furthermore, a thin-layer chromatographic monitoring on C18 bonded silica gel with ion-pairing allowed the monitoring of the ionizable metabolites for substrates that were demonstrated to decompose most rapidly. These complementary analyses showed that the sensitivity of the highly sensitive strain towards cymoxanil was related to the disappearance of cyano-oximes studied from culture medium, namely to the ability of the strain B. cinerea to metabolize them. PMID:11936693

Tellier, Frédérique; Fritz, René; Leroux, Pierre; Carlin-Sinclair, Abel; Cherton, Jean-Claude



Chromatography Theory  

NSDL National Science Digital Library

James Hardy has produced a number of presentation-style lecture modules for analytical chemistry. This is a very good module that covers the introductory theory of chromatography. The material includes information on chromatography along with plate and kinetic theories. It is a colorful and well planned presentation of the introduction to chromatography that could be used as the basis for lectures on chromatography at the introductory analytical chemistry or instrumental analysis level.


Affinity Chromatography  

NSDL National Science Digital Library

This is an experiment showing the application of affinity chromatography to the separation of albumin from horse serum. A brief introduction of affinity chromatography and how it is being used in this specific experiment is given. This appears to be a good experiment to show the advantages of affinity chromatography in separating specific proteins from a complex matrix and would be useful in a biochemistry course or a course that is specifically looking at differing types of chromatography.

DiResta, Dan


ELSEVIER Journal of Chromatography A, 693 (1995) 197-203 CHROMATOGRAPHYA  

E-print Network

ELSEVIER Journal of Chromatography A, 693 (1995) 197-203 JOURNALOF CHROMATOGRAPHYA Adsorptive interaction of Ficoll standards with porous glass size-exclusion chromatography columns Gita Shah, Paul L in chromatographic and porous media. Calibrating porous glass size-exclusion chromatography columns with Ficoll, we

Dubin, Paul D.


Chemical analysis of uranium and titanium niobotantalate metamict minerals by ion-exchange chromatography and spectrophotometric procedures.  


An ion-exchange separation followed by spectrophotometric determinations is applied to some metamict minerals. These minerals, containing very high amounts of elements which present some problems to the analyst, such as uranium, titanium, niobium and rare-earth elements, are fused with potassium bisulphate, and the cooled melts dissolved in sulphuric acid. The solutions are passed through a series of three ion-exchange columns to separate those mineral-forming elements for which the colorimetric procedures suffer interference from the elements listed above. The procedure has been tested with a synthetic solution and with solutions of the minerals. PMID:18963564

Mazzucotelli, A; Vannucci, R; Vannucci, S; Passaglia, E



The simultaneous determination of active ingredients in cough-cold mixtures by isocratic reversed-phase ion-pair high-performance liquid chromatography.  


A simple, rapid and accurate method for the simultaneous determination of active ingredients in cough-cold mixtures using isocratic reversed-phase ion-pair high-performance liquid chromatography has been developed. It involves the use of an octadecylsilane column as the stationary phase with methanol, water, tetrahydrofuran, phosphoric acid mixtures as mobile phase including sodium dioctylsulphosuccinate as the ion-pair agent. The pH of the mobile phase was adjusted to 4.6 by means of phosphoric acid and ammonium hydroxide solutions. The proposed method involves the simple dilution of the samples with the mobile phase and the addition of metoclopramide hydrochloride as the internal standard. The active ingredients under investigation were chlorpheniramine, codeine, diphenhydramine, ephedrine, ethylmorphine, phenylephrine, phenylpropanolamine and pholcodine, which exist as various combinations in cough-cold mixtures. The optimum composition of the mobile phase and the optimum flow rate were determined and are reported. The method was applied to the determination of active ingredients in seven commercially available cough-cold mixtures. PMID:2577452

Lau, O W; Chan, K; Lau, Y K; Wong, W C



Simultaneous determination of imidacloprid and carbendazim in water samples by ion chromatography with fluorescence detector and post-column photochemical reactor.  


A new analytical method has been developed and validated for the simultaneous determination of pesticides from different classes using ion chromatography-online photochemical derivatisation-fluorescence detector (IC-hv-FD). Fluorimetric detection was performed at ?ex/?em=332 nm/367 nm for imidacloprid and then detector was set at ?ex/?em=247 nm/470 nm for carbendazim. The two pesticides imidacloprid and carbendazim were successfully separated isocratically on an IonPac(®) AS11 (250 mm × 4 mm i.d; 13 µm particle size, Dionex) anion-exchange column using 40 mM KOH with 10% (v/v) acetonitrile and pumped at a flow rate of 1.0 mL min(-1). Under the optimized conditions, the limit of detection (LOD, S/N=3) of imidacloprid and carbendazim were 7.8 µg L(-1) and 67 µg L(-1), respectively. The experimental results showed that there was good linearity with a correlation coefficient (r)?0.9966 over the range of 0.05-10 mg L(-1) for imidacloprid and 0.2-15 mg L(-1) for carbendazim. Good reproducibility with a relative standard deviation (RSD, n=7) less than 4.5%. Finally, the proposed method was applied with satisfactory results to the analysis of these pesticides in ground water, lake water and river water without any pre-treatment of samples. The average spiked recoveries were in the range of 90-104%. PMID:24148383

Subhani, Qamar; Huang, Zhongping; Zhu, Zuoyi; Zhu, Yan



Monitoring of phosphorus oxide ion for analytical speciation of phosphite and phosphate in transgenic plants by high-performance liquid chromatography-inductively coupled plasma mass spectrometry.  


Large amounts of phosphate fertilizers utilized in agriculture and their relatively poor efficiency are of high ecological and economic concern. Therefore, transgenic plants capable of metabolizing phosphite are being engineered. In support of this biotechnological task, analytical speciation of phosphorus in biological tissues is required. In this study, plant extracts were analyzed by liquid chromatography-inductively coupled plasma mass spectrometry at m/z of elemental phosphorus and phosphorus oxide ions. Using polymeric-based anion exchange column and millimolar concentration of nitric acid in potassium phthalate mobile phase (pH 2.5), phosphite and phosphate ions were baseline resolved with retention times 6.95 ± 0.03 and 7.90 ± 0.03 min and with a total chromatographic run time 10 min. The detection limits were 1.58 and 1.74 ?g P L(-1) at m/z 47, as compared to 2.18 and 2.04 ?g P L(-1) at m/z 31, respectively. The results obtained in real world samples for the two detection modes were in good agreement, yet signal acquisition at m/z 47 enabled better precision without collision/reaction cell (RSD below 2%) as compared to RSD around 4% obtained at m/z 31 using He-pressurized cell (3.5 mL min(-1)). PMID:23782169

Torres Elguera, Julio Cesar; Yañez Barrientos, Eunice; Wrobel, Katarzyna; Wrobel, Kazimierz



Determination of trace levels of herbicides and their degradation products in surface and ground waters by gas chromatography/ion-trap mass spectrometry  

USGS Publications Warehouse

A rapid, specific and highly sensitive method is described for the determination of several commonly used herbicides and their degradation products in surface and ground waters by using gas chromatography/ion-trap mass spectrometry. The compounds included atrazine, and its degradation products desethylatrazine and desisopropylatrazine; Simazine; Cyanazine; Metolachlor; and alachlor and its degradation products, 2-chloro-2', 6'-diethylacetanilide, 2-hydroxy-2', 6'-diethylacetanilide and 2,6-diethylaniline. The method was applied to surface-water samples collected from 16 different stations along the lower Mississippi River and its major tributaries, and ground-water samples beneath a cornfield in central Nebraska. Average recovery of a surrogate herbicide, terbuthylazine, was greater than 99%. Recoveries of the compounds of interest from river water spiked at environmental levels are also presented. Full-scan mass spectra of these compounds were obtained on 1 ng or less of analyte. Data were collected in the full-scan acquisition mode. Quantitation was based on a single characteristic ion for each compound. The detection limit was 60 pg with a signal-to-noise ratio of greater than 10:1.

Pereira, W.E.; Rostad, C.E.; Leiker, T.J.



[Determination of ten basic dyes in meat products by ultra fast liquid chromatography-ion trap time of flight mass spectrometry].  


A method was developed for the simultaneous determination of 10 basic dyes in meat products using ultra fast liquid chromatography-ion trap time of flight mass spectrometry (LC-IT-TOF-MS). The target analytes were separated at a flow rate of 0.2 mL/min on a Waters Acquity UPLC BEH C18 (100 mm x 2.1 mm, 1.7 microm) column with a gradient elution. The mobile phase was 5 mmol/L ammonium acetate-acetonitrile (containing 0.1% (v/v) formic acid). The identification and quantification were achieved in positive ion mode with electro spray ionization source. The samples were extracted with a simple procedure using acetonitrile and cleaned up by weak cation exchange (Oasis WCX) solid phase extraction column. Ten basic dyes were determined by LC-IT-TOF-MS, and quantified by external standard method. The developed method showed a good linearity over the wide range of 1.0 - 100.0 microg/L, and the relative standard deviations (n = 7) were less than 8.54%. The average recoveries of the ten basic dyes at three levels (2, 10 and 25 microg/kg) were ranged from 65.39% to 119.18%. Therefore, this method, owing to its simplicity, rapidity and high sensitivity, has a good applicability to the simultaneous determination of dye residues in meat products. PMID:23256378

Zhang, Donglei; Wang, Lina; Chen, Xiaozhen; Wang, Jin; Cao, Hui; Huang, Liying



An integrated analytical approach using ion chromatography, PIXE and electron microscopy to point out the differences in composition of PM10 individual particles  

NASA Astrophysics Data System (ADS)

This study presents a part of a project aiming at characterizing the PM10 composition in the Venice area with detailed chemical analyses using various techniques. The concentrations of six major inorganic ions (Cl-, NO3-, SO42-, Na+, NH4+, Mg2+) and 19 elements (Na, Mg, Al, Si, P, S, Cl, K, Ca, Ti, V, Cr, Mn, Fe, Ni, Cu, Zn, Br, Pb) were quantified using ion chromatography and PIXE, respectively. The masses of PM10 daily samples and their chemical contents were studied in relationship to micro-meteorological parameters to select a few of them characterized by very different chemical profiles. Four samples from the whole period were categorized as representative of i) clean days; ii) sea spray generation events; iii) high contribution of mineral dust and iv) heavy pollution events. Individual particle analyses of the samples were also performed by SEM-EDS microscopy. Six different classes of particles were identified (mineral particles, chlorides, sulfates, elemental and organic carbon compounds, metals and biological particles) and an estimation of their abundance yielded a significant relationship between the chemical content and composition in individual particles of PM10. Further information was also obtained on PM10 source contributions, morphology, mineralogy and mixed state of particles demonstrating the importance of combining different analytical approaches.

Masiol, Mauro; Ceccato, Daniele; Squizzato, Stefania; Carturan, Sara; Pavoni, Bruno



Quantitative Phosphoproteome Analysis of Lysophosphatidic Acid Induced Chemotaxis applying Dual-step ¹?O Labeling Coupled with Immobilized Metal-ion Affinity Chromatography  

SciTech Connect

Reversible protein phosphorylation is a central cellular regulatory mechanism in modulating protein activity and propagating signals within cellular pathways and networks. Development of more effective methods for the simultaneous identification of phosphorylation sites and quantification of temporal changes in protein phosphorylation could provide important insights into molecular signaling mechanisms in a variety of different cellular processes. Here we present an integrated quantitative phosphoproteomics approach and its applications for comparative analysis of Cos-7 cells in response to lysophosphatidic acid (LPA) gradient stimulation. The approach combines trypsin-catalyzed 16O/18O labeling plus 16O/18O-methanol esterification labeling for quantitation, a macro- Immobilized Metal-ion Affinity Chromatography trap for phosphopeptide enrichment, and a monolithic capillary column with integrated electrospray emitter. LC separation and MS/MS is followed by neutral loss-dependent MS/MS/MS for phosphopeptide identification using a linear ion trap (LTQ)-FT mass spectrometer and complementary searching algorithms for interpreting MS/MS spectra. Protein phosphorylation involved in various signaling pathways of cell migration were identified and quantified, such as mitogen-activated protein kinase 1, dual-specificity mitogen-activated protein kinase kinase 2, and dual-specificity tyrosine-phosphorylation regulated kinase 1b, and a number of Rho GTPase-activating proteins. These results demonstrate the efficiency of this quantitative phosphoproteomics approach and its application for rapid discovery of phosphorylation events associated with gradient sensing and cell chemotaxis.

Ding, Shi-Jian; Wang, Yingchun; Jacobs, Jon M.; Qian, Weijun; Yang, Feng; Tolmachev, Aleksey V.; Du, Xiuxia; Wang, Wei; Moore, Ronald J.; Monroe, Matthew E.; Purvine, Samuel O.; Waters, Katrina M.; Heibeck, Tyler H.; Adkins, Joshua N.; Camp, David G.; Klemke, Richard L.; Smith, Richard D.



¹³C labelled internal standards--a solution to minimize ion suppression effects in liquid chromatography-tandem mass spectrometry analyses of drugs in biological samples?  


Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is frequently used to identify and quantify drugs in human biological samples due to the high selectivity and sensitivity of this technique. However, ion suppression effects caused by co-eluting compounds: drugs, metabolites, matrix components, impurities and degradation products, are a major concern. Stable isotope labelled internal standards (SIL ISs), usually deuterium ((2)H) labelled, are often used to compensate for these effects. In many LC separations the retention times of (2)H labelled ISs and their analogues will differ. Ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) is increasingly being used for bio-analysis. With the better chromatographic resolution provided with sub 2 ?m particles, larger separation between analytes and their (2)H labelled analogues can be expected, which might reduce the benefits of the SIL IS. There is a greater difference in physico-chemical properties between hydrogen isotopes than between isotopes of other elements. (13)C, (15)N and (18)O labelled ISs are more similar to their analytes than (2)H labelled ISs and thereby expected to behave more similarly in chromatographic separations. In this study we have investigated the use of (13)C and (2)H labelled ISs for the determination of amphetamine and methamphetamine by UPLC-MS/MS. The (13)C labelled ISs were co eluting with their analytes under different chromatographic conditions while the (2)H labelled ISs and their analytes were slightly separated. An improved ability to compensate for ion suppression effects were observed when the (13)C labelled ISs were used. Furthermore, an UPLC-MS/MS method for determination of amphetamine and methamphetamine in urine using (13)C labelled ISs has been developed and validated. Unfortunately, there are few (13)C labelled ISs commercial available today. If more (13)C labelled ISs become commercial available they may well be the coming solution to minimize ion suppression/enhancement effects in LC-MS/MS analyses of drugs in biological samples. PMID:22119139

Berg, Thomas; Strand, Dag Helge



Field detection of bacillus spore aerosols with stand-alone pyrolysis-gas chromatography and ion mobility spectrometry  

Microsoft Academic Search

A commercially available, hand-held chemical vapor detector was modified to detect Gram-positive Bacillus subtilis var. globigii spores (BG) in outdoor field scenarios. An Airborne Vapor Monitor (AVM) ion mobility spectrometry (IMS) vapor detector was interfaced to a biological sample processing and transfer introduction system. The biological sample processing was accomplished by quartz tube pyrolysis (Py), and the resultant vapor was

A. P. Snyder; Waleed M. Maswadeh; John A. Parsons; Ashish Tripathi; Henk L. Meuzelaar; Jacek P. Dworzanski; Man-Goo Kim



Liquid chromatography-mass spectrometry with electron impact and fast ion bombardment with a ribbon storage interface  

Microsoft Academic Search

The design and performance of a new moving ribbon liquid chromatograph-mass spectrometer interface for use with secondary ion mass spectrometry (SIMS) and conventional electron impact ionizaton (EI) are described and initial results are presented. The interface includes a 120 cm long region at atmospheric pressure and aerosol deposition of the LC effluent to allow the complete evaporation of the LC

R. D. Smith; J. E. Burger; A. L. Johnson




Microsoft Academic Search

Distribution coefficients of tetravalent titanium and zirconium between ; ion-exchange resins and hydrochloric acid solutions of various concentrations ; were determined. It was found that there is no sorption of the mentioned ; elements from 0.1 to 6 N hydrochloric acid solutions with anion exchange resins. ; A method was developed for the quantitative chromatographic separation of ; titanium and

T. A. Belyavskaya; I. P. Alimarin; I. F. Kolosova



Determination of dimethyl fumarate (DMFu) in silica gel pouches using gas chromatography coupled ion trap mass spectrometry  

Microsoft Academic Search

A simple method for the determination of dimethyl fumarate (DMFu) in silica gel pouches has been developed. The gas chromatographic behaviour of DMFu was investigated; the instrumental method, based on EI mass spectrometry coupled with an Ion Trap Detector operating whether in full scan mode or in MS\\/MS mode was also investigated. Several factors have been evaluated and optimized during

Patrizia Stefanelli; Danilo Attard Barbini; Silvana Girolimetti; Angela Santilio; Roberto Dommarco



Ion-paired extraction of cephalosporins in acetone prior to their analysis by capillary liquid chromatography in environmental water and meat samples.  


Ion-pair extraction of cephalosporins from aqueous solution into acetone by the addition of ammonium sulfate to a 1:2 (v/v) acetone-water solvent was carried out followed by their determination using reversed-phase capillary liquid chromatography. The analytes included are cephoperazone, cefquinome, cephalexin, cephapirin, cephaloniun, cephamandole, cephazolin and cephadroxile. In order to form the ion-pair, hexadecyltrimethylammonium bromide (CTAB) was selected as cationic ion-pairing agent at a concentration of 0.9 mM using 10mM phosphate buffer at pH 8 as the optimum condition for the aqueous solution. The applied methodology, named salting-out assisted liquid/liquid extraction (SALLE) involves the use of 1.25 g of ammonium sulfate as salting-out agent. The separation of cephalosporins using a Luna C18 (150 mm × 0.3mm, 5 µm, 100 Å) column was achieved under the following conditions: a gradient program combining solvent A (0.1% formic acid in water, pH 4) and solvent B (acetonitrile-methanol (50:50, v/v)), at a flow rate of 20 µl min(-1), column temperature 35°C and injection volume 7 µl with UV detection at 250 nm. The limits of quantification for the studied compounds were between 4.3 and 22.7 ?g/L for water samples and 4.1 and 73.3 ?g/kg in the case of beef samples, lower than the maximum residue limits permitted by the EU for this kind of food. The developed methodology has demonstrated its suitability for the analysis of these widely applied antibiotics in environmental water and meat samples, including beef and pork muscle, with high sensitivity, precision and satisfactory recoveries. PMID:24054686

Quesada-Molina, Carolina; García-Campaña, Ana M; del Olmo-Iruela, Monsalud



Qualitative Metabolome Analysis of Human Cerebrospinal Fluid by 13C-/12C-Isotope Dansylation Labeling Combined with Liquid Chromatography Fourier Transform Ion Cyclotron Resonance Mass Spectrometry  

NASA Astrophysics Data System (ADS)

Metabolome analysis of human cerebrospinal fluid (CSF) is challenging because of low abundance of metabolites present in a small volume of sample. We describe and apply a sensitive isotope labeling LC-MS technique for qualitative analysis of the CSF metabolome. After a CSF sample is divided into two aliquots, they are labeled by 13C-dansyl and 12C-dansyl chloride, respectively. The differentially labeled aliquots are then mixed and subjected to LC-MS using Fourier-transform ion cyclotron resonance mass spectrometry (FTICR MS). Dansylation offers significant improvement in the performance of chromatography separation and detection sensitivity. Moreover, peaks detected in the mass spectra can be readily analyzed for ion pair recognition and database search based on accurate mass and/or retention time information. It is shown that about 14,000 features can be detected in a 25-min LC-FTICR MS run of a dansyl-labeled CSF sample, from which about 500 metabolites can be profiled. Results from four CSF samples are compared to gauge the detectability of metabolites by this method. About 261 metabolites are commonly detected in replicate runs of four samples. In total, 1132 unique metabolite ion pairs are detected and 347 pairs (31%) matched with at least one metabolite in the Human Metabolome Database. We also report a dansylation library of 220 standard compounds and, using this library, about 85 metabolites can be positively identified. Among them, 21 metabolites have never been reported to be associated with CSF. These results illustrate that the dansylation LC-FTICR MS method can be used to analyze the CSF metabolome in a more comprehensive manner.

Guo, Kevin; Bamforth, Fiona; Li, Liang



Comprehensive Two-Dimensional Separation of Hydroxylated Polybrominated Diphenyl Ethers by Ultra-Performance Liquid Chromatography Coupled with Ion Mobility-Mass Spectrometry  

NASA Astrophysics Data System (ADS)

A comprehensive two-dimensional system coupling ultra-performance liquid chromatography (UPLC) and ion mobility-mass spectrometry (IM-MS) has been applied for the separation and analysis of hydroxylated polybrominated diphenyl ethers (OH-PBDEs). A complex mixture containing 23 OH-PBDE congeners ranging from hydroxylated monobromodiphenyl ether (OH-monoBDE) to hydroxylated octabromodiphenyl ether (OH-octaBDE) was satisfactorily separated within 16 min of analysis time. The first-dimensional reversed-phase UPLC was performed on a sub-2 ?m BEH C18 chromatographic column using acetonitrile-water gradient elution program with a flow rate ramp. It enabled excellent chromatographic separation for both between-class and within-class OH-PBDEs based on their differences in hydrophobicity. Following the pre-ionization resolution in the first dimension, the second-dimensional IM-MS employed a hybrid electrospray quadrupole ion mobility time-of-flight mass spectrometer and added an extra post-ionization separation for between-class OH-PBDE congeners on account of their relative mobility disparity during a very short period of 8.80 ms. The orthogonality of the developed two-dimensional system was evaluated with the correlation coefficient of 0.9665 and peak spreading angle of 14.87°. The peak capacity of the system was calculated to be approximately 2 and 15 times higher than that of the two dimensions used alone, respectively. The two-dimensional separation plane also contributed to the removal of background interference ions and the enhanced confidence in the characterization of OH-PBDEs of interest.

Ma, Qiang; Wang, Chao; Bai, Hua; Xi, Hai-Wei; Xi, Guang-Cheng; Ren, Xiao-Min; Yang, Yu; Guo, Liang-Hong



Characterization of a rapid and reliable method for iodide biomonitoring in serum and urine based on ion chromatography-ICP-mass spectrometry.  


An appropriate and controlled supply of thyroid hormones is vital for proper body function. In turn, an appropriate synthesis of T3 and T4 in the thyroid gland is dependent on a sufficient and balanced iodide concentration in blood serum. Due to widespread iodine deficiency or some cases of iodine over exposure, iodide biomonitoring in serum is important and it is that biomonitoring approach being closest to the bioavailable I(-) supply for the thyroid gland. Therefore, this paper describes a biomonitoring method for iodide determination in serum based on ion chromatography-inductively coupled plasma mass spectrometry (IC-ICP-MS). Since in literature only very few data are available on iodide in serum but many in urine the method is also extended to I(-) monitoring in urine. The method was additionally designed to have short analysis time (8min) for increased sample throughput, good precision in serial measurement (serum: 4.86%; urine: 1.4%), and day-to-day determination (serum: 5.7%; urine: 2.28%), high accuracy (serum: 105%; urine: 101%) and good recovery (serum: 102%; urine: 99%) even in matrix-rich samples at low I(-) concentration. Also, investigations were performed to elucidate whether internal standardization during chromatography, sample preparation for protein-matrix removal or matrix-matched calibration are advantageous for analytical performance. Finally, limits of detection (3?) of 0.12?g/L or 0.05?g/L (serum or urine) and limit of quantification (10?) of 0.39?g/L or 0.17?g/L (serum or urine) were achieved. PMID:24933092

Michalke, Bernhard; Witte, Heidi



Isolation of ?-linolenic acid biohydrogenation products by combined silver ion solid phase extraction and semi-preparative high performance liquid chromatography.  


Polyunsaturated fatty acids typically found in cattle feed include linoleic (LA) and ?-linolenic acid (ALA). In the rumen, microbes metabolize these resulting in the formation of biohydrogenation products (BHP), which can be incorporated into meat and milk. Bioactivities of LA-BHP, including conjugated linoleic acid (cis (c) 9,trans (t) 11-18:2 and t10,c12-18:2) and trans fatty acid isomers (t9-, t10- and t11-18:1) have been investigated, but effects of several BHP unique to ALA have not been extensively studied, and most ALA-BHP are not commercially available. The objective of the present research was to develop methods to purify and collect ALA-BHP using silver ion (Ag(+)) chromatography in sufficient quantities to allow for convenient bioactivity testing in cell culture. Fatty acid methyl esters (FAME) were prepared from perirenal adipose tissue from a cow enriched with ALA-BHP by feeding flaxseed. These were applied to Ag(+)-solid phase extraction, and eluted with hexane with increasing quantities of acetone (1, 2, 10, 20%) or acetonitrile (2%) to pre-fractionate FAME based on degree of unsaturation and double bond configuration. Fractions were collected, concentrated and applied to semi-preparative Ag(+)-high performance liquid chromatography (HPLC) for the isolation and collection of purified isomers, which was accomplished using isocratic elutions with hexane containing differing amounts of acetonitrile (from 0.015 to 0.075%). Purified trans-18:1 isomers collected ranged in purity from 88 to 99%. Purity of the ALA-BHP dienes collected, including c9,t13-18:2, t11,c15-18:2 and t10,c15-18:2, exceeded 90%, while purification of other dienes may require the use of other complementary procedures (e.g. reverse phase HPLC). PMID:25579113

Turner, T D; Meadus, W J; Mapiye, C; Vahmani, P; López-Campos, Ó; Duff, P; Rolland, D C; Church, J S; Dugan, M E R



Determination of linear alkylbenzenesulfonates and their degradation products in soils by liquid chromatography-electrospray-ion trap multiple-stage mass spectrometry.  


Linear alkylbenzenesulfonates (LAS) (C(10)-C(13)) and their degradation products, sulfophenyl carboxylate compounds (SPCs) (C(2)-C(6), C(8), C(11)), have been extracted from soil samples with methanol, isolated, concentrated by solid-phase extraction, and determined by liquid chromatography/negative ion electrospray quadrupole ion-trap tandem mass spectrometry (MS(n)). The ion fragmentation processes and pathways were studied in detail by MS, MS(2), and MS(3). Upon collision-induced dissociation, the deprotonated molecules of LASs render the ethylene-substituted benzenesulfonate ion (m/z 183), the fragmentation of which gave the intense signal at m/z 119, corresponding to the ethylene-substituted phenoxide ion formed by the loss of sulfur dioxide. The fragmentation pattern of SPCs shows that, for the analytes of large carbon atom chains (>5C), the neutral loss of water is favored whereas for those of short carbon atoms chain, the loss of carbon dioxide is more frequent. Multiple reaction monitoring using isolation only for MS and using isolation and fragmentation for MS(2) and MS(3) were used to identify and quantify each compound. The three MS modes have been validated in terms of sensitivity, selectivity, and precision, showing that each MS stage used reduces sensitivity 10 times. Recoveries from soil were higher than 65% at LOQ level for all the analytes tested, except for C(2)-C(4) SPCs by any MS mode, with relative standard deviation lower than 19%. The utility of the method is demonstrated by successfully quantifying real samples treated with these products. Quantification limits for the methodology developed in this work ranged from 0.5 to 50 microg kg(-1) by MS, from 2 to 400 microg kg(-1) by MS(2), and from 20 to 4000 microg kg(-1) by MS(3). Concentration levels of LASs and SPCs-ranging from 0.1 to 15 mg kg(-1)-were found in soil samples amended with sludges, thus indicating their input and persistence in the soil compartment. PMID:15144200

Andreu, Vicente; Picó, Yolanda



Nano-fabricated size exclusion chromatograph  

NASA Technical Reports Server (NTRS)

This paper describes the development of a nano-fabricated size exclusion chromatograph (nSEC) based on the principle that molecules traveling through amicrocolumn containing nano-fabricated features will have characteristic elution times that directly correlate to molecular weight. Compared to conventional size exclusion chromatography, the nSEC offers greater control over the size exclusion process; mass fabrication; integration of the separation column with associated valves, pumps, and detectors; and dramatic reductions in instrument mass and power requirements.

Svehla, D.; Feldman, S.; Feldman, J.; Grunthaner, F.; Shakkottai, P.; Castillo, L. del; White, V.



Simultaneous determination of artificial sweeteners, preservatives, caffeine, theobromine and theophylline in food and pharmaceutical preparations by ion chromatography  

Microsoft Academic Search

A novel ion chromatographic method was proposed for the simultaneous determination of artificial sweeteners (sodium saccharin, aspartame, acesulfame-K), preservatives (benzoic acid, sorbic acid), caffeine, theobromine and theophylline. The separation was performed on an anion-exchange analytical column operated at 40°C within 45 min by an isocratic elution with 5 mM aqueous NaH2PO4 (pH 8.20) solution containing 4% (v\\/v) acetonitrile as eluent,

Qing-Chuan Chen; Jing Wang



Express analysis of explosives, chemical warfare agents and drugs with multicapillary column gas chromatography and ion mobility increment spectrometry  

Microsoft Academic Search

Description of a gas chromatograph designed for express analysis of explosives (2,4-dinitrotoluene, 2,4,6-trinitrotoluene, pentaerythritol tetranitrate), chemical warfare agents (mustard gas, lewisite, sarin) and drugs (heroin, cocaine hydrochloride, crack) is given. The devices comprises a multicapillary chromatographic column and an ion mobility increment spectrometer (MCC–IMIS). The main analytical characteristics of an IMIS (estimated detection limit (DL), linear dynamic range (LDR), speed

Igor A. Buryakov



Determination of inorganic anions by capillary ion-exchange chromatography using polyethylenimine-coated octadecyl-bonded phases.  


The utility of a cationic polymer polyethylenimines (PEI) for coating some commercially available silica-based stationary phases packed in capillaries of 0.32 mm i.d. and 10 cm long was studied for the separation and direct UV detection of inorganic anions (iodate, bromate, nitrite, bromide, and nitrate ions). With a super-endcapped octadecylated silica stationary phase, which yielded the best separation in terms of resolution and retention time of individual anions, the effect of the pH and composition of the eluent on the elution behavior of individual anions was studied. The relative standard deviations of the retention time, peak area and peak height for 60 successive injections (running time of more than 10 h) were not more than 1.32, 2.30 and 1.94, respectively, with the exceptions of the peak area and peak height of the bromate ion. The detection limits at 210 nm ranged from 1 to 7 ppm. This method was successfully applied to determine concentrations of nitrate ions present in pills and beverage samples. PMID:22729052

Sedyohutomo, Anang; Suzuki, Hiroshi; Fujimoto, Chuzo



Development of a methodology utilizing gas chromatography ion-trap tandem mass spectrometry for the determination of low levels of caffeine in surface marine and freshwater samples.  


A methodology for monitoring low level of caffeine in aqueous samples via gas chromatography coupled with an ion-trap tandem mass spectrometry detection system (IT-MS/MS) was developed. Four IT-MS/MS operating parameters, including the collision-induced dissociation (CID) voltage, the excitation time (ET), the isolation time (IT) and the maximum ionization time (MIT) were optimized in order to maximize the sensitivity of the IT-MS/MS technique towards the analyte and its isotope-labeled standard. After optimization, a limit of detection of 500 fg microl(-1) with S/N = 3 was achieved. Taking into account blank values and the matrix background, a method detection limit of 1.0-2.0 ng l(-1) was derived and applied to all of the samples analyzed in the study. Various mass spectrometric conditions have been applied to caffeine and its trimethyl-(13)C-labeled standard to elucidate fragmentation pathways for new and commonly occurring product ions observed in the collision-induced dissociation (CID) spectra produced by the ion trap. Ion structures and fragmentation pathway mechanisms have been proposed and compared with previously published data. An isotope dilution method using (13)C-labeled caffeine as a surrogate internal standard was employed to determine and correct the recovery of native caffeine in water samples. The developed methodology has been applied for the determination of caffeine in surface marine and freshwater samples collected on the west coast of Vancouver Island in British Columbia, Canada. The results obtained for the marine water samples indicated a wide variation in the level of caffeine, ranging from 4.5 to 149 ng l(-1), depending on the location of the sampling site within the inlet. The concentrations of caffeine in samples from lakes associated with various residential densities ranged from ND to 6.5, 1.8 to 10.4 and 6.1 to 21.7 ng l(-1) for low, moderate and high residential densities, respectively. PMID:18535819

Verenitch, Sergei S; Mazumder, Asit



Study of anthocyanic profiles of twenty-one hybrid grape varieties by liquid chromatography and precursor-ion mass spectrometry.  


The anthocyanins of 21 hybrid red varieties produced by crossing V. vinifera, V. riparia, V. labrusca, V. lincecumii and V. rupestris species, the profiles for which have not yet been reported, were studied. Profiles were determined by LC/DAD, and identification of single anthocyanins was confirmed by LC/MS precursor-ion analysis. Anthocyanidin precursors (pelargonidin at m/z 271, dephinidin at m/z 303, cyanidin at m/z 287, petunidin at m/z 317, peonidin at m/z 301, and malvidin at m/z 331) and precursors of monoglucoside compounds allowed 24 different compounds to be identified. Analysis of precursor ions of monoglucoside anthocyanins at low capillary voltage revealed the signals of diglucosides only, providing a very selective method for analysis of diglucoside anthocyanins in grape. According to anthocyanin profile, the samples were subdivided into two groups: one characterized by the substantial presence of diglucoside compounds (particularly Seyve Villard 23-399 and Seyve Villard 23-369) and one by the scarce presence or practical absence of diglucosides (Seibel 10878, Burdin 4077, and Galibert 238-35). Particularly interesting for producing anthocyanin for the natural colorant industry were the varieties Siebel 8357, Bacò 30-12 and Terzi 100-31. PMID:22688043

De Rosso, M; Tonidandel, L; Larcher, R; Nicolini, G; Ruggeri, V; Dalla Vedova, A; De Marchi, F; Gardiman, M; Flamini, R



Mechanistic insights on chaotropic interactions of liophilic ions with basic pharmaceuticals in polar ionic mode liquid chromatography.  


We report for the first time the effect of liophilic mobile phase additives on the mechanism of chiral recognition of basic chiral pharmaceutical on a vancomycin based chiral stationary phase (CSP). Using methanol as the mobile phase and 0.005% formic acid as pH modifier, we evaluated the effect of different concentrations of three types of liophilic anions, formate (HCOO(-)), nitrate (NO3(-)), and acetate (CH3COO(-)), on enantioresolution (Rs), enantioselectivity (?) and retention factor (k) of enantiomers of fluoxetine and atenolol. The effect of liophilic ion types on k followed the Hofmeister series: CH3COO(-)>HCOO(-)>NO3(-). Increasing concentration from 4 to 20mM resulted in decreases in Rs and k in accordance to hydrophobicity of the liophilic anion. The effect of temperature or mobile phase composition on enantioseparation was determined at 13-40°C. For both analytes, standard changes in enthalpy (?H°) and entropy (?S°) calculated using van't Hoff plots (lnk against 1/T) to varied from -4.99 to -0.63 kJ/mol and -11.82 to 9.47 J/mol, respectively. The van't Hoff plots showed elution order of the enantiomers of each analyte did not reverse in the temperature range studied. Chiral recognition of the enantiomers of atenolol and fluoxetine in the presence of liophilic ions was enthalpy driven. PMID:25311487

Sanganyado, Edmond; Lu, Zhijiang; Gan, Jay



Autonomous, waste-free eluent generation and suppression in a single device: electrodialytic eluent reflux for ion chromatography.  


Eluent reflux provides a new approach to suppress and reflux (recover) eluent without the continuous generation of chromatographic waste. The current work utilized a device containing ion exchange membranes at the electrodes, in order to prohibit electrolysis gases from entering the eluent stream. Two resin beds (separated by a membrane stack) were responsible for suppressing incoming eluent and regenerating the suppressed eluent to nearly its original concentration after detection. A greater than expected dilution in the eluent concentration was observed as a result of the minor leakage of potassium ions through the anion membrane stack into the electrode chamber. The incomplete recovery of the eluent was offset by the addition of a three port valve (DRV) to regulate eluent concentration. Over 48 h of continuous operation (192 injections), the device's performance was stable (RSD of 0.21% with the three port valve, compared to RSD 3.73% without). The device was able to operate for up to four weeks using 1L of eluent. Chromatograms showing the reproducibility of the device are presented for anions. PMID:24401425

Elkin, Kyle R; Riviello, John M



Simultaneous determination of diclofenac, its human metabolites and microbial nitration/nitrosation transformation products in wastewaters by liquid chromatography/quadrupole-linear ion trap mass spectrometry.  


An analytical method was developed and validated for the first determination of five major human metabolites of the non-steroidal anti-inflammatory drug diclofenac as well as two microbial transformation products in wastewater. The method was based on the extraction of diclofenac and the chemically synthetized compounds by solid-phase extraction (SPE), using a hydrophilic-lipophilic balanced polymer followed by liquid chromatography (LC) coupled to hybrid quadrupole-linear ion trap mass spectrometry (QqLIT-MS). Quantitation was performed by the internal standard approach, to correct for matrix effects. The accuracy of the method was generally higher than 40% for raw and treated wastewater with a precision below 12%. In wastewater influent and effluent samples the detection limits for the majority of target compounds were 0.3-2.5ngL(-1) and 0.1-3.1ngL(-1), respectively. The method was applied to the analysis of influent and effluent wastewater samples from urban wastewater treatment plants. Moreover, to obtain an extra tool for confirmation and identification of the studied diclofenac-derived compounds, Information-Dependent Acquisition (IDA) experiments were performed, with selected reaction monitoring (SRM) as the survey scan and an enhanced product ion (EPI) scan as the dependent scan. Diclofenac and its major human metabolite, 4'-hydroxydiclofenac were detected in all samples at concentrations of 331-1150ngL(-1) and 585-6000ngL(-1), respectively. Neither microbial transformation product of diclofenac was detected in any of the influent samples analyzed, but in effluents, their concentrations ranged from 4 to 105ngL(-1). PMID:24835592

Osorio, Victoria; Imbert-Bouchard, Marta; Zonja, Bozo; Abad, José-Luis; Pérez, Sandra; Barceló, Damià



Rapid and sensitive gas chromatography ion-trap mass spectrometry method for the determination of tobacco specific N-nitrosamines in secondhand smoke  

SciTech Connect

Tobacco-specific nitrosamines (TSNAs) are some of the most potent carcinogens in tobacco and cigarette smoke. Accurate quantification of these chemicals is needed to help assess public health risks. We developed and validated a specific and sensitive method to measure four TSNAs in both the gas- and particle-phase of secondhand smoke (SHS) using gas chromatography and ion-trap tandem mass spectrometry,. A smoking machine in an 18-m3 room-sized chamber generated relevant concentrations of SHS that were actively sampled on Teflon coated fiber glass (TCFG) filters, and passively sampled on cellulose substrates. A simple solid-liquid extraction protocol using methanol as solvent was successfully applied to both filters with high recoveries ranging from 85 to 115percent. Tandem MS parameters were optimized to obtain the best sensitivity in terms of signal to-noise ratio (S/N) for the target compounds. For each TSNA, the major fragmentation pathways as well as ion structures were elucidated and compared with previously published data. The method showed excellent performances with a linear dynamic range between 2 and 1000 ng mL-1, low detection limits (S/N> 3) of 30-300 and precision with experimental errors below 10percent for all compounds. Moreover, no interfering peaks were observed indicating a high selectivity of MS/MS without the need for a sample clean up step. The sampling and analysis method provides a sensitive and accurate tool to detect and quantify traces of TSNA in SHS polluted indoor environments.




Quantifying the Labeling and the Levels of Plant Cell Wall Precursors Using Ion Chromatography Tandem Mass Spectrometry1[W][OA  

PubMed Central

The biosynthesis of cell wall polymers involves enormous fluxes through central metabolism that are not fully delineated and whose regulation is poorly understood. We have established and validated a liquid chromatography tandem mass spectrometry method using multiple reaction monitoring mode to separate and quantify the levels of plant cell wall precursors. Target analytes were identified by their parent/daughter ions and retention times. The method allows the quantification of precursors at low picomole quantities with linear responses up to the nanomole quantity range. When applying the technique to Arabidopsis (Arabidopsis thaliana) T87 cell cultures, 16 hexose-phosphates (hexose-Ps) and nucleotide-sugars (NDP-sugars) involved in cell wall biosynthesis were separately quantified. Using hexose-P and NDP-sugar standards, we have shown that hot water extraction allows good recovery of the target metabolites (over 86%). This method is applicable to quantifying the levels of hexose-Ps and NDP-sugars in different plant tissues, such as Arabidopsis T87 cells in culture and fenugreek (Trigonella foenum-graecum) endosperm tissue, showing higher levels of galacto-mannan precursors in fenugreek endosperm. In Arabidopsis cells incubated with [U-13CFru]sucrose, the method was used to track the labeling pattern in cell wall precursors. As the fragmentation of hexose-Ps and NDP-sugars results in high yields of [PO3]?/or [H2PO4]? ions, mass isotopomers can be quantified directly from the intensity of selected tandem mass spectrometry transitions. The ability to directly measure 13C labeling in cell wall precursors makes possible metabolic flux analysis of cell wall biosynthesis based on dynamic labeling experiments. PMID:20442274

Alonso, Ana P.; Piasecki, Rebecca J.; Wang, Yan; LaClair, Russell W.; Shachar-Hill, Yair



Liquid chromatography tandem mass spectrometric quantitation of sulfamethazine and its metabolites: direct analysis of swine urine by triple quadrupole and by ion trap mass spectrometry.  


This work describes a new method for the quantitation of trace amounts of sulfamethazine (SMZ) and its main metabolite, N4-acetylsulfamethazine (Ac-SMZ), in swine urine, using high-performance liquid chromatography (HPLC) tandem mass spectrometric analysis of crude urine after addition of internal standard and simple dilution with water. The aim was to determine whether residues of this sulfamidic drug, normally administered to swine in order to prevent infectious diseases, were present in urine at levels lower than those permitted by regulatory authorities before human consumption (EU Project SMT, contract number CT 96-2092). A 10 microL volume of diluted urine was injected into a very short, narrow-bore chromatographic column (Zorbax SB-C18 2.1 i. d. x30 mm length, 3.5 microm pore size). Elution of the analytes of interest was achieved in less than seven minutes using a rapid gradient (from 20 to 80% methanol in 3 minutes). Either a PE Sciex API 365 triple quadrupole (QqQ), operated in the selected reaction monitoring (SRM) mode, or a Finnigan LCQ ion trap (IT) mass spectrometer, operated in narrow-range product ion scan, was used as the final detector. Electrospray (ESI) was used as the ionization technique. A comparison of the two tandem mass spectrometers was performed by analyzing the same set of test samples, at three concentration levels, on three different days. Linearity of responses of the calibration standards, intra- and inter-assay precision of the samples, specificity and limits of detection were evaluated for both systems. Both the QqQ and the IT instrument was suitable for rapid, sensitive and specific determination of the analytes, although the overall performance of the QqQ was slightly superior in terms of linearity, precision and sensitivity. PMID:10844733

Bartolucci, G; Pieraccini, G; Villanelli, F; Moneti, G; Triolo, A



Evaluation of the extent of heterogeneity in the Glycera dibranchiata monomer haemoglobin fraction by the use of n.m.r. and ion-exchange chromatography.  

PubMed Central

The coelomic haemoglobin of Glycera dibranchiata is known to be separable into monomeric and higher-Mr fractions. Although exhibiting homogeneity with respect to Mr, the extent of haemoglobin heterogeneity for the monomer fraction has never been adequately assayed. In the present paper we demonstrate that there exists in the monomer haemoglobin fraction reproducibly detectable heterogeneity regardless of the presence or absence of proteinase inhibitors during the isolations. These results show that, considered on the same time scale as previous preparations used for amino acid sequencing, crystallography and kinetics, the monomer haemoglobin fraction is highly heterogeneous. Application of ion-exchange chromatography and ion-filtration methods resulted in the isolation of four resolvable haem protein components from the Glycera monomer haemoglobin fraction. Three of these components were isolated in sufficient quantity to employ proton n.m.r. as a successful analytical tool for discriminating the individual haemoglobins. These results are not surprising. Several previous studies indicated less extensive heterogeneity in the monomer fraction. Moreover, the ability of the Glycera monomer haemoglobin to bind oxygen at even quite low partial pressures has been attributed to functional diversity originating in multiple haemoglobin components. The present work reveals the extent of the haemoglobin heterogeneity. The results show that it is more extensive than previously believed. Examination of this monomer fraction is particularly important, since crystallography indicates that one of the components of the monomer fraction lacks the E-7 (distal) histidine residue. As a consequence, the identification of such extensive heterogeneity is important to many previously published ligand-binding studies. PMID:3977860

Kandler, R L; Constantinidis, I; Satterlee, J D



Determination of seven arsenic species in seafood by ion exchange chromatography coupled to inductively coupled plasma-mass spectrometry following microwave assisted extraction: method validation and occurrence data.  


The determination of seven arsenic species in seafood was performed using ion exchange chromatography on an IonPac AS7 column with inductively coupled plasma mass spectrometry detection after microwave assisted extraction. The effect of five parameters on arsenic extraction recoveries was evaluated in certified reference materials. The recoveries of total arsenic and of arsenic species with the two best extraction media (100% H(2)O and 80% aqueous MeOH) were generally similar in the five seafood certified reference materials considered. However, because MeOH co-elutes with arsenite, which would result in a positively biased arsenite concentration, the 100% H(2)O extraction conditions were selected for validation of the method. Figures of merit (linearity, LOQs (0.019-0.075 mg As kg(-1)), specificity, trueness (with recoveries between 82% (As(III)) and 104% (As(V) based on spikes or certified concentrations), repeatability (3-14%), and intermediate precision reproducibility (9-16%) of the proposed method were satisfactory for the determination of arsenite, monomethylarsonic acid, dimethylarsinic acid, arsenate, arsenobetaine and arsenocholine in fish and shellfish. The performance criteria for trimethylarsine oxide, however, were less satisfactory. The method was then applied to 65 different seafood samples. Arsenobetaine was the main species in all samples. The percentage of inorganic arsenic varied between 0.4-15.8% in shellfish and 0.5-1.9% at the utmost in fish. The main advantage of this method that uses only H(2)O as an extractant and nitric acid as gradient eluent is its great compatibility with the long-term stability of both IEC separation and ICP-MS detection. PMID:21147319

Leufroy, Axelle; Noël, Laurent; Dufailly, Vincent; Beauchemin, Diane; Guérin, Thierry



[Screening and confirmation of carcinogenic dyes in textiles by high performance liquid chromatography-linear ion trap/orbitrap high resolution mass spectrometry].  


A method of high performance liquid chromatography-linear ion trap/Orbitrap high resolution mass spectrometry (HPLC-LTQ/Orbitrap MS) was used to screen and confirm carcinogenic dyes in textiles. The analytes were extracted from textile samples with pyridine/water (1/1, v/v) in a water bath under controlled conditions (95 degrees C, 150 r/min), and then filtered with a 0.22 microm polytetrafluoroetylene (PTFE) membrane. The eluates were separated on a CAPCELL PAK C18 column (100 mm x 2.0 mm, 5 microm) using gradient elution with acetonitrile/ 5 mmol/L ammonium acetate aqueous solution containing 0.01% formic acid (in positive mode) and acetonitrile/5 mmol/L ammonium acetate aqueous solution (in negative mode), and finally detected by HPLC-LTQ/Orbitrap MS in ESI modes. Full scan experiments were performed over the range of m/z 200-800. The screening and quantitative analysis were carried out by the accurate mass of quasi-molecular ion and the peak area in extracted chromatogram with accurate mass, respectively. The confirmatory analysis for target compounds was performed with the retention time and qualitative fragments obtained by data-dependent scan mode. Under the optimal conditions, nine carcinogenic dyes were routinely detected with mass accuracy below 5 x 10(-6) (5 ppm), and good linearities were provided in their respective linear ranges with correlation coefficients higher than 0.99. The limits of detection were in the range of 0.125-25 mg/kg. The average recoveries at three spiked levels were in the range of 62.13%116.28% with the relative standard deviations (RSDs) lower than 15%. The proposed method was applied to screen and confirm the nine carcinogenic dyes in textile samples. It is convenient and reliable. PMID:24432638

Xiu, Xiaoli; Luo, Xin; Niu, Zengyuan; Ye, Xiwen; Zhixu, Tang; Li, Jingying; Wang, Yongwei; Du, Wei



Ion Torrent-based transcriptional assessment of a Corynebacterium pseudotuberculosis equi strain reveals denaturing high-performance liquid chromatography a promising rRNA depletion method  

PubMed Central

Corynebacterium pseudotuberculosis equi is a Gram-positive pathogenic bacterium which affects a variety of hosts. Besides the great economic losses it causes to horse-breeders, this organism is also known to be an important infectious agent to cattle and buffaloes. As an outcome of the efforts in characterizing the molecular basis of its virulence, several complete genome sequences were made available in recent years, enabling the large-scale assessment of genes throughout distinct isolates. Meanwhile, the RNA-seq stood out as the technology of choice for comprehensive transcriptome studies, which may bring valuable information regarding active genomic regions, despite of the still impeditive associated costs. In an attempt to increase the use of generated reads per instrument run, by effectively eliminating unwanted rRNAs from total RNA samples without relying on any commercially available kits, we applied denaturing high-performance liquid chromatography (DHPLC) as an alternative method to assess the transcriptional profile of C.?pseudotuberculosis. We have found that the DHPLC depletion method, allied to Ion Torrent sequencing, allows mapping of transcripts in a comprehensive way and identifying novel transcripts when a de novo approach is used. These data encourage us to use DHPLC in future transcriptional evaluations in C.?pseudotuberculosis. PMID:23316806

Castro, Thiago L P; Seyffert, Nubia; Ramos, Rommel T J; Barbosa, Silvanira; Carvalho, Rodrigo D O; Pinto, Anne Cybelle; Carneiro, Adriana Ribeiro; Silva, Wanderson Marques; Pacheco, Luis G C; Downson, Christopher; Schneider, Maria P C; Miyoshi, Anderson; Azevedo, Vasco; Silva, Artur



Development of cadmium/silver/palladium separation by ion chromatography with quadrupole inductively coupled plasma mass spectrometry detection for off-line cadmium isotopic measurements.  


A separation method was investigated to perform off-line cadmium isotopic measurements on a (109)Ag transmutation target. Ion chromatography (IC) with Q ICPMS detection (quadrupole inductively coupled plasma mass spectrometry detection) was chosen to separate cadmium from the isobarically interfering elements, silver and palladium, present in the sample. The optimization of chromatographic conditions was particularly studied. Several anion and cation columns (Dionex AG11(®), CS10(®) and CS12(®)) were compared with different mobile phases (HNO(3), HCl). The separation procedure was achieved with a carboxylate-functionalized cation exchange CS12 column using 0.5 M HNO(3) as eluent. The developed technique yielded satisfactory results in terms of separation factors (greater than 5) and provides an efficient solution to obtain rapidly purified cadmium fractions (decontamination factors higher 100,000 for silver and palladium) which can directly be analyzed by multi collection inductively coupled plasma mass spectrometry (MC ICPMS). By applying the proposed procedure, accurate and precise cadmium isotope ratios were determined for the irradiated (109)Ag transmutation target. PMID:21703628

Gautier, C; Bourgeois, M; Isnard, H; Nonell, A; Stadelmann, G; Goutelard, F



Carbohydrate Signatures of Aquatic Macrophytes and Their Dissolved Degradation Products as Determined by a Sensitive High-Performance Ion Chromatography Method  

PubMed Central

The sugar contents of emergent macrophytes from a freshwater lake, a freshwater swamp, and a salt marsh in the southeastern United States were examined together with the dissolved free sugars produced during macrophyte degradation and in natural water samples collected adjacent to macrophyte stands. Simultaneous separation of up to 13 neutral and 2 amino sugars together with 3 uronic acids and muramic acid was achieved by anion-exchange high-performance ion chromatography. As little as 10 pmol or a concentration of 20 nM sugar can be detected by pulsed amperometry, a greater sensitivity for sugar quantification than that of previously reported detection techniques used in conjunction with either gas or liquid chromatographic systems. Optimum conditions for hydrolysis of plant material by using trifluoroacetic acid were determined, and internal standards were used to quantify losses due to matrix effects and solid-phase extraction of samples. Our data demonstrate that ratios of certain indicator sugars in undegraded macrophytes differ significantly from ratios of dissolved free sugars formed during macrophyte degradation, reflecting the complex processes (biological and physical) involved in vascular plant degradation in aquatic ecosystems. Natural water samples collected adjacent to macrophyte beds contained dissolved free sugars at concentrations of 620 nM (lake), 890 nM (freshwater swamp), and 2,300 nM (salt marsh). Sugar signatures of these natural water samples were similar to those of macrophyte degradation products. PMID:16348579

Wicks, Richard J.; Moran, Mary Ann; Pittman, Laura J.; Hodson, Robert E.



Analysis of anthelmintics in surface water by ultra high performance liquid chromatography coupled to quadrupole linear ion trap tandem mass spectrometry.  


A method based on ultra high performance liquid chromatography coupled to quadrupole linear ion trap mass spectrometry (UHPLC-QqLIT-MS) has been developed to investigate occurrence of 10 anthelmintic drugs from different structural groups (moxidectin, flubendazole, fenbendazole, levamisol, mebendazole, oxibendazole, albendazole, triclabendazole, febantel and praziquantel) in surface water. Analytes were pre-concentrated by solid phase extraction (SPE) using hydrophilic-lipophilic polymeric based sorbent. Quantification of investigated analytes was done using deuterated compounds as internal standards in order to minimize matrix effect. Analyte recoveries from spiked samples at two concentration levels were above 75% for most of the analytes. The main advantages of developed method are fast separation using UHPLC and therefore short analysis time, combined with good sensitivity which is demonstrated by low ngL(-1) detection limits. The developed method was applied for analysis of anthelmintics in the Llobregat River (NE Spain) and its main tributaries (rivers Anoia and Cardener). Eight out of ten anthelmintics were detected in all analyzed samples with the concentrations in low ngL(-1) level. The method fills the gap on analytical methodologies for determination of anthelmintic drugs in the environment. PMID:24289978

Zrn?i?, Mirta; Gros, Meritxell; Babi?, Sandra; Kaštelan-Macan, Marija; Barcelo, Damia; Petrovi?, Mira



Determination of aminophenols and phenol in hair colorants by ultrasound-assisted solid-phase dispersion extraction coupled with ion chromatography.  


A simple and reliable ultrasound-assisted solid-phase dispersion extraction coupled with ion chromatography was developed for the determination of aminophenols and phenol. The highly viscous hair colorant was dispersed in solvents using anhydrous sodium sulfite having dual functions of dispersant and antioxidant. The use of anhydrous sodium sulfite did not change the sample volume because it could completely dissolve in solution after matrix dispersion. The extraction and cleanup were combined in one single step for simplifying operation. The extraction process could be rapidly accomplished within 9 min with high sample throughput under the synergistic effects of vibration, ultrasound, and heating. Satisfactory linearity was observed with correlation coefficients higher than 0.9992, and the limits of detection varied from 0.02 to 0.09 mg/L. The applicability of the proposed method was demonstrated by measuring the concentrations of aminophenols and phenol in 32 different commercial hair color products. The recoveries ranged from 86.4-101.2% with the relative standard deviations in the range of 0.52-4.3%. The method offers an attractive alternative for the analysis of trace phenols in complex matrices. PMID:24841051

Zhong, Zhixiong; Li, Gongke; Wu, Rong; Zhu, Binghui; Luo, Zhibin



Effect of Photoperiod on the Levels of Endogenous Gibberellins in Spinach as Measured by Combined Gas Chromatography-selected Ion Current Monitoring 1  

PubMed Central

The changes in the levels of five endogenous gibberellins (GAs) in spinach in relation to photoperiodic treatment have been examined by combined gas chromatography-selected ion current monitoring. Long-day treatment caused a 5-fold decline in the level of GA19 while the levels of GA20 and GA29 increased dramatically during the same period. In absolute terms, the level of GA20 increased from 0.8 microgram per 100 grams dry weight in short days to 5.5 micrograms per 100 grams dry weight after 14 long days. The levels of GA17 and GA44 did not change significantly with long-day treatment. These results are consistent with the hypothesis that GA19 is converted to GA20 and that this conversion is under photoperiodic control. Since stem growth in spinach is correlated with an increase in the level of GA20, one major aspect of photoperiodic control of stem growth might be the availability of GA20 through regulation of the conversion of GA19 to GA20. PMID:16661539

Metzger, James D.; Zeevaart, Jan A. D.



A rare haemoglobin variant (Hb Phnom Penh) manifesting as a falsely high haemoglobin A1c value on ion-exchange chromatography.  


Most haemoglobin (Hb) variants are clinically silent. However, some Hb variants may interfere with the measurement of haemoglobin A1c (HbA1c), resulting in spurious values depending on the assays used. We herein report the case of a 53-year-old Taiwanese man with type 2 diabetes mellitus, who presented with an abnormal HbA1c peak on ion-exchange chromatography. Additional investigations, including intensified self-monitored blood glucose tests, an alternative HbA1c assay, and a glycaemic indicator based on a different method, revealed that the HbA1c values were falsely elevated. Subsequent DNA analysis confirmed that the patient was heterozygous for the insertion of an isoleucine residue at codons 117/118 of the a1-globin gene, Hb Phnom Penh. Clinical laboratorians should be aware of the interfering factors in their HbA1c analysis. Cautious inspection of the chromatogram may provide a valuable clue to the presence of an Hb variant. PMID:25189312

Chen, Chi-Fen; Tai, Yen-Kuang



Determination of pesticides associated with suspended sediments in the San Joaquin River, California, USA, using gas chromatography-ion trap mass spectrometry  

USGS Publications Warehouse

An analytical method useful for the quantification of a range of pesticides and pesticide degradation products associated with suspended sediments was developed by testing a variety of extraction and cleanup schemes. The final extraction and cleanup methods chosen for use are suitable for the quantification of the listed pesticides using gas chromatography-ion trap mass spectrometry and the removal of interfering coextractable organic material found in suspended sediments. Methylene chloride extraction followed by Florisil cleanup proved most effective for separation of coextractives from the pesticide analytes. Removal of elemental sulfur was accomplished with tetrabutylammonium hydrogen sulfite. The suitability of the method for the analysis of a variety of pesticides was evaluated, and the method detection limits (MDLs) were determined (0.1-6.0 ng/g dry weight of sediment) for 21 compounds. Recovery of pesticides dried onto natural sediments averaged 63%. Analysis of duplicate San Joaquin River suspended-sediment samples demonstrated the utility of the method for environmental samples with variability between replicate analyses lower than between environmental samples. Eight of 21 pesticides measured were observed at concentrations ranging from the MDL to more than 80 ng/g dry weight of sediment and exhibited significant temporal variability. Sediment-associated pesticides, therefore, may contribute to the transport of pesticides through aquatic systems and should be studied separately from dissolved pesticides.

Bergamaschi, B.A.; Baston, D.S.; Crepeau, K.L.; Kuivila, K.M.



A new method for the determination of carbonyl compounds in wines by headspace solid-phase microextraction coupled to gas chromatography-ion trap mass spectrometry.  


A new analytical method for the determination of 18 carbonyl compounds [2,3-pentadione, hexanal, (E)-2-hexen-1-al, octanal, acetoin, (E)-2-octenal, furfural, decanal, (E)-2-nonenal, benzaldehyde, 5-methylfurfural, (E,E)-2-cis-6-nonadienal, ?-damascenone, phenylacetaldehyde, acetophenone, (E,E)-2,4-decadienal, benzophenone, and vanillin] in wines using automated headspace solid-phase microextraction (HS/SPME) coupled to gas chromatography-ion trap mass spectrometry (GC-ITMS) was developed. Five fibers with different polarities were tested, and a study of the influence of various factors such as time and extraction temperature, desorption time and temperature, pH, and ionic strength and content in tannins, anthocyans, sucrose, SO(2), and alcoholic degree was conducted. These factors were optimized using a synthetic wine doped with the different analytes. The proposed method affords wide ranges of linearity, good linearity (r(2) > 0.998), values of repeatability and reproducibility lower than 5.5% of RSD, and detection limits ranging from 0.62 ?g/L for ?-damascenone to 129.2 ?g/L for acetoin. Therefore, the optimized method was applied to the quantitative analysis of the aforementioned analytes in real samples of wines. PMID:21121610

Pérez Olivero, Sergio J; Pérez Trujillo, Juan P