title: Rapid determination of branched chain amino acids in human blood plasma by pressure-assisted capillary electrophoresis with contactless conductivity detection.
abstract: A CE method with contactless conductivity detection has been developed for the clinical determination of the branched chain amino acids (BCAAs) valine, isoleucine and leucine in human blood plasma. The CE separation was performed in an optimised BGE with composition of 3.2 M acetic acid in 20% v/v methanol, pH 2.0. The achieved separation time was 125 s when using a capillary with an effective length of 14.7 cm, electric field intensity of 0.96 kV/cm and simultaneous application of a hydrodynamic pressure of 50 mbar. The separation efficiency in blood plasma equalled 461 000 theoretical plates/m for valine and isoleucine, and 455 000 theoretical plates/m for leucine; the detection limits are equal to 0.4 μM for all three amino acids. The RSD values for repeatability of the migration time equalled 0.1% for measurements during a single day and 0.3% for measurements on different days; the RSD values for repeatability of the peak areas equalled 2.3-2.6% for measurements during a single day and 2.7-4.6% for measurements on different days. It followed from the performed tests that the plasmatic levels of BCAAs attain a maximum 60 min after intravenous application of an infusion of BCAAs.
authors: Institute of Biochemistry, Cell and Molecular Biology, Third Faculty of Medicine, Charles University in Prague, Prague, Czech Republic.
authors: Petr
authors: P
authors: Tůma
authors: 2nd Internal Department of Third Faculty of Medicine and Faculty Hospital Královské Vinohrady, Centre for Research on Diabetes, Metabolism and Nutrition, Charles University in Prague, Prague, Czech Republic.
authors: Jan
authors: J
authors: Gojda
mesh_terms: D000328:Adult; D000597:Amino Acids, Branched-Chain / Q000097:blood* / Q000493:pharmacokinetics; D000068456:Clinical Studies as Topic; D003920:Diabetes Mellitus; D004553:Electric Conductivity; D019075:Electrophoresis, Capillary / Q000379:methods*; D005260:Female; D006801:Humans; D007328:Insulin / Q000097:blood / Q000378:metabolism; D057230:Limit of Detection; D016014:Linear Models; D015203:Reproducibility of Results
chemical_list: D000597:Amino Acids, Branched-Chain; D007328:Insulin
keywords: Amino acids; CE; Clinical analysis; Contactless conductivity detection; Diabetes mellitus