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Determination of glucuronic acid in serum by HPLC.
Title: Determination of glucuronic acid in serum by HPLC.
Author: Yuko HANAOKA; Kazuhiro SHIKAKUME; Yasusuke MATSUMOTO; Yasukuni YAKABE; Yukihiro TARUI; Yoshikatsu SAYAMA (Chemical Standards Department, Tokyo Laboratory, Chemicals Inspection and Testing Institute; Chemical Assessment Center, Chemicals Inspection and Testing Institute; Product Development Department, Personal Health Care Division, Chugai Pharmaceutical Co., Ltd.)
Source: Bunseki kagaku; ISSN:0525-1931; VOL.47; NO.2; PAGE.119-125; (1998)
Abstract: An analytical method for the determination of glucuronic acid in human serum was developed by high-performance liquid chromatography (HPLC) using an ion-pair method, and the optimum analytical conditions were investigated. This method involves the acidic degradation of 200 μl of human serum with 4 M sulfuric acid, neutralization with sodium hydroxide, a reaction with 0.5 M 3-metyl-1-phenyl-5-pyrazolone (PMP) methanol, and neutralization with 0.4 M sulfuric acid. Excess reaction regent was extracted with chloroform, and an aliquot of the aqueous solution was injected to the HPLC. By using an ion-pair reagent, glucuronic acid and glucose in human serum could be separated. Under the optimized analytical conditions, the calibration curve was linear over the range 550 mg/l. The recoveries of glucuronic acid from human serum samples at a level of 550 mg/l were 8294%. The intra-day and inter-day precision of the method were 0.64.3% (n = 5) and 4.36.9% (n = 5), respectively. The concentration of glucuronic acid in human serum was constant for 30 days at -20°C. This method was applied to the determination of glucuronic acid in human serum after single oral administration of glucuronolactone to volunteers.
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