DETERMINATION OF VITAMIN-D3 AND 25-HYDROXYVITAMIN-D3 IN SERA BY COLUMN-SWITCHING HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY WITH FLUORESCENCE DETECTION

被引：16

作者：

IWATA, T ^{[1
]}

YAMAGUCHI, M ^{[1
]}

HANAZONO, H ^{[1
]}

IMAZATO, Y ^{[1
]}

NAKAMURA, M ^{[1
]}

OHKURA, Y ^{[1
]}

机构：

[1] KYUSHU UNIV 62,FAC PHARMACEUT SCI,FUKUOKA 812,JAPAN

来源：

ANALYTICAL SCIENCES | 1990年 / 6卷 / 03期

关键词：

3,4-dihydro-6,7-dimethoxy-4-methyl-3-oxoquinoxaline-2-carbonyl azide; column-switching high performance liquid chromatography; fluorescence detection; human serum; rat serum; Vitamin D[!sub]3[!/sub]25-hydroxyvitamin D[!sub]3[!/sub;

D O I：

10.2116/analsci.6.361

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

A column-switching high performance Liquid chromatographic method with fluorescence detection for the determination of vitamin Dj and 25-hydroxyvitamin D in human and rat sera is described. The vitamins in a lipid extract from serum, obtained by solid-phase extraction technique using a Bond-Elut C18 cartridge, are converted with 3,4-dihydro-6,7-dimethoxy-4-methyl-3-oxoquinoxaline-2-carbonyl azide into the corresponding fluorescent derivatives. The derivatives are separated from endogenous interfering substances by column-switching chromatography. The chromatographic system consisted of a precolumn chromatography for clean-up of the derivatives and an analytical column chromatography for the complete separation of the derivatives. The derivatives are detected fluorometrically at excitation and emission wavelengths of 360 and 440 nm, respectively. The detection limits (S/N=3) for vitamin D3 and 25-hydroxyvitamin D3 are 15 and 8 fmol, respectively, in a 10μl injection volume. The sensitivity permits simultaneous determination of the vitamins in 1 ml of normal human and rat sera. © 1990, The Japan Society for Analytical Chemistry. All rights reserved.

引用

页码：361 / 366

页数：6

共 29 条

[1] QUANTITATION OF THE MAIN METABOLITES OF VITAMIN-D IN A SINGLE SERUM SAMPLE .1. EXTRACTION, SEPARATION AND PURIFICATION OF METABOLITES [J].