MICRODETERMINATION OF HYALURONIC-ACID IN HUMAN URINE BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY

被引：10

作者：

AKIYAMA, H ^{[1
]}

TOYODA, H ^{[1
]}

YAMANASHI, S ^{[1
]}

SAGEHASHI, Y ^{[1
]}

TOIDA, T ^{[1
]}

IMANARI, T ^{[1
]}

机构：

[1] CHIBA UNIV,FAC PHARMACEUT SCI,1-33 YAYOI CHO,CHIBA 260,JAPAN

来源：

BIOMEDICAL CHROMATOGRAPHY | 1991年 / 5卷 / 05期

关键词：

D O I：

10.1002/bmc.1130050502

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A high performance liquid chromatographic (HPLC) system is described for determination of the unsaturated disaccharide (DELTA-Di-HA) derived from hyaluronic acid (HA) in human urine by digestion with hyaluronidase SD. The effects of eluents on the separation of DELTA-Di-HA and DELTA-Di-0S, which is derived from the reaction of chondroitin with the enzyme, have been studied. The established chromatographic conditions were as follows-column: a stainless steel tube (4 mm i.d. x 250 mm) packed with TSKgel NH2-60; eluent: a mixture of acetonitrile and 0.1 M Tris-HCl buffer containing 0.1 M boric acid and 10 mM sodium sulphate, pH 7.0 (64:36, v/v). The strong fluorescence of unsaturated disaccharide after the reaction with 2-cyanoacetamide in alkaline medium was used for post-column detection. The calibration curve for DELTA-Di-HA was linear in the range 5 pmol-5 nmol with a practical detection limit of 2 pmol. The assay coefficients of variation (n = 5) at 200 pmol for DELTA-Di-HA and DELTA-Di-0S were 1.7 and 1.5%, respectively. This HPLC system has been applied to the determination of HA in human urine.

引用

页码：189 / 192

页数：4

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