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Cm Effective Length (cm + effective_length)
Selected AbstractsEffects of lactate and acetate on the determination of serum ethyl glucuronide by CZEELECTROPHORESIS, Issue 23 2006Michaela Mrázková Abstract The analysis of ethyl glucuronide,(EtG), a marker of recent alcohol consumption, in serum with an optimized CZE assay is reported. The method uses a 0.1-mm,id fused-silica capillary of 50,cm effective length that is coated with linear polyacrylamide, a pH,4.4 nicotinic acid/,-aminocaproic acid (EACA) BGE, reversed polarity and indirect analyte detection. The assay is based on a 1:1 dilution of serum with deionized water and has LODs for EtG, lactate and acetate of 3.8×10,7,M, 2.60×10,6,M and 2.18×10,6,M, respectively. Separation of EtG from endogenous macro- and microcomponents (anionic serum components of high and low concentration, respectively) and its quantification are shown to be possible for a wide range of lactate (stacker) and acetate (destacker) concentrations, macrocomponents that have an impact on the CZE behavior of EtG and that change after intake of ethanol. The assay has been successfully applied to the analysis of EtG, lactate and acetate in (i),sera of volunteers that ingested known amounts of alcohol and (ii),samples of patients that were classified (teetotalers and social drinkers vs. alcohol abusers) via analysis of carbohydrate-deficient transferrin. [source] Development of off-line and on-line capillary electrophoresis methods for the screening and characterization of adenosine kinase inhibitors and substratesELECTROPHORESIS, Issue 12 2006Jamshed Iqbal Abstract Fast and convenient CE assays were developed for the screening of adenosine kinase,(AK) inhibitors and substrates. In the first method, the enzymatic reaction was performed in a test tube and the samples were subsequently injected into the capillary by pressure and detected by their UV absorbance at 260,nm. An MEKC method using borate buffer (pH,9.5) containing 100,mM SDS (method,A) was suitable for separating alternative substrates (nucleosides). For the CE determination of AMP formed as a product of the AK reaction, a phosphate buffer (pH,7.5 or 8.5) was used and a constant current (95,,A) was applied (method,B). The methods employing a fused-silica capillary and normal polarity mode provided good resolution of substrates and products of the enzymatic reaction and a short analysis time of less than 10,min. To further optimize and miniaturize the AK assays, the enzymatic reaction was performed directly in the capillary, prior to separation and quantitation of the product employing electrophoretically mediated microanalysis (EMMA, method,C). After hydrodynamic injection of a plug of reaction buffer (20,mM Tris-HCl, 0.2,mM MgCl2, pH,7.4), followed by a plug containing the enzyme, and subsequent injection of a plug of reaction buffer containing 1,mM,ATP, 100,,M adenosine, and 20,,M,UMP as an internal standard,(I.S.), as well as various concentrations of an inhibitor, the reaction was initiated by the application of 5,kV separation voltage (negative polarity) for 0.20,min to let the plugs interpenetrate. The voltage was turned off for 5,min (zero-potential amplification) and again turned on at a constant current of ,60,,A to elute the products within 7,min. The method employing a polyacrylamide-coated capillary of 20,cm effective length and reverse polarity mode provided good resolution of substrates and products. Dose,response curves and calculated Ki values for standard antagonists obtained by CE were in excellent agreement with data obtained by the standard radioactive assay. [source] Capillary electrophoretic chiral separation of hydroxychloroquine and its metabolites in the microsomal fraction of liver homogenatesELECTROPHORESIS, Issue 5-6 2006Carmem Dickow Cardoso Abstract A rapid, selective, and low-cost chiral capillary electrophoretic method was developed for the simultaneous analysis of hydroxychloroquine (HCQ) and its three chiral metabolites: desethylchloroquine (DCQ), desethylhydroxychloroquine (DHCQ), and bisdesethylchloroquine (BDCQ) in the microsomal fraction of liver homogenates. After liquid,liquid extraction using toluene as extracting solvent, the drug and metabolites were resolved on a fused-silica capillary (50,,m ID, 50,cm total length, and 42,cm effective length), using 100,mmol/L of Tris/phosphate buffer, pH,9.0 containing 1% w/v sulfated-,-CD and 30,mg/mL hydroxypropyl-,-CD. Detection was carried out at 220,nm. The extraction procedure was efficient in removing endogenous interferents, and low values (,15%) for CVs and deviation from theoretical values were demonstrated for both within-day and between-day assays. The quantitation limit was 125,ng/mL with linear response over the 125,2000,ng/mL of concentration range for all metabolites. After validation, the method was used for an in vitro metabolism study of HCQ. The major HCQ metabolite formed by microsomal enzymes was (,)-(R)-DHCQ. [source] Fast CE analysis of adrenergic amines in different parts of Citrus aurantium fruit and dietary supplementsJOURNAL OF SEPARATION SCIENCE, JSS, Issue 16 2010Laura Mercolini Abstract A CE method has been developed for the simultaneous analysis of the adrenergic amines synephrine, octopamine and tyramine in Citrus aurantium (bitter orange) fruit extracts and in dietary supplements. The analytes were separated on a fused silica capillary (50,,m id, 40.0,cm effective length, 48.5,cm total length) using a BGE composed of phosphate buffer (pH 2.5, 50,mM) and applying a 30,kV potential. The samples were injected hydrodynamically at 50,mbar for 25,s. The use of photodiode array detection (,=195,nm) allowed the quantification of the analytes and the control of peak purity. The method has been fully validated, obtaining satisfactory values of precision and extraction yield. The analytes are extracted with water from the dried whole fruits or fruit parts (endocarp, mesocarp and exocarp) or from the commercial formulations and directly injected into the CE apparatus. The results obtained were satisfactory in terms of precision (RSD <,5.7%) and accuracy (recovery >,89%). Thus, the method has demonstrated to be suitable for the qualitative and quantitative determination of synephrine, octopamine and tyramine in C. aurantium extracts, for dietary supplement quality control and for food adulteration identification. [source] Analysis of gastrodin and tetramethylpyrazine in traditional Chinese preparations by micellar electrokinetic chromatographyJOURNAL OF SEPARATION SCIENCE, JSS, Issue 6-7 2003Wang Rongying Abstract A simple, rapid, and accurate micellar electrokinetic chromatographic method has been developed for determination of gastrodin and tetramethylpyrazine in three traditional Chinese preparations: Zhennaoning jiaonang, Yangxue shengfa jiaonang, and Xiaoshuan zaizao wan. Running buffer comprising 50 mM sodium tetraborate and 15 M sodium dodecylsulfate (SDS), pH 9.50, was found to be most suitable for the separation. All experiments was performed with a 47 cm (40 cm effective length)×75 ,m ID uncoated fused-silica capillary and UV detection at 200 nm. The linear calibration ranges were 2.5,200 ,g mL,1 (R = 0.999) for gastrodin and 5.0,200 ,g mL,1 (R = 0.997) for tetramethylpyrazine; the detection limits were 0.5 ,g mL,1 and 0.8 ,g mL,1, respectively. Recoveries of the two analytes from the samples, calculated by use of a method described in detail in the text, were between 94.21 and 104.46%. The amounts of gastrodin and tetramethylpyrazine in the preparations were easily determined within 10 min. [source] Micellar electrokinetic capillary chromatography of methylxanthines-containing beverages: discussion of the molecular species involvedJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 4 2005Alicia B Pomilio Abstract Micellar electrokinetic capillary chromatography (MECC) experimental conditions were applied to 12 samples of methylxanthine-containing infusions of different commercial brands of yerba mate, coffee, tea and cocoa as well as two cola drinks. The best resolution in this mode of automated high-performance capillary electrophoresis (HPCE) was achieved here when using 15 kV voltage in an uncoated fused-silica capillary of 45 cm length (40 cm effective length), 50 mM sodium dodecylsulfate, 90 mM pH 8.5 borate buffer and UV detection. Theobromine, caffeine and theophylline were separated, and the peak splitting due to tautomeric species was observed. Experimental conditions were controlled, keeping constant the size of the elution window in each analysis. The limit of detection was less than 1 mg l,1, the limit of quantitation was 2.5 mg l,1 and the work range was 2.5,300 mg l,1. This HPCE,MECC system has proved suitable for the analysis/quality control of xanthines in beverages for consumption. Roles of various parameters as well as distinctly charged species of each xanthine and the origin of peak splitting in this MECC system are discussed. Copyright © 2004 Society of Chemical Industry [source] | ||||||||||