Japanese Journal of Clinical Chemistry
Online ISSN : 2187-4077
Print ISSN : 0370-5633
ISSN-L : 0370-5633
A Method for the Determination of γ-Carboxyglutamic Acid in Human Bile by High-Performance Liquid Chromatography with Fluorometric Detection
Shigeru Notsumoto, Masahito Higashiura, Yuji Usui, Kunio Kobayashi
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Keywords: γ-carboxyglutamicacid, peptide-bound γ-carboxyglutamic acid, high-performance liquid chromatography, fluorometric detection, columnswitching system
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1998 Volume 27 Issue 4 Pages 239-244

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Abstract
We established a novel method to determine γ-carboxyglutamic acid (Gla) inhumanbile, which has been suggested to be related to gallstone formation. Gla was derived with o-phthalaldehyde, then measured by the HPLC method with fluorometric detection (Ex 345nm, Em 445nm) and a column-switching system. Peptidebound Gla was effectively converted into free Gla by heating with 11mol/l potassium hydroxide at 110°C for 20 hrs. The calibration curve for Gla was linear over the range 76 to 7600pmol/ml. The detection limit of Gla was 4pmol/ml. Accuracy and precision (CV) were 98% (mean recovery) and 1.9-4.0% (n=4 each), respectively.
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