SIMULTANEOUS DETERMINATION OF RETINOL, ALPHA-TOCOPHEROL, AND 25-HYDROXYVITAMIN-D IN HUMAN SERUM BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY

被引:38
作者
AKSNES, L
机构
[1] Department of Pediatrics, University of Bergen, Haukeland Hospital, Bergen
关键词
VITAMIN-A; VITAMIN-D; VITAMIN-E; RETINOL; 25-HYDROXYCHOLECALCIFEROL; ALPHA-TOCOPHEROL; HUMAN SERUM; HPLC;
D O I
10.1097/00005176-199404000-00015
中图分类号
R57 [消化系及腹部疾病];
学科分类号
摘要
Determination of the serum or plasma levels of retinol (vitamin A), 25-hydroxyvitamin D, and cr-tocopherol (vitamin E) are the most frequently used parameters to evaluate status of vitamin A, D, and E, and also to assess the gastrointestinal absorption of the vitamins. We present a simple and sensitive method for simultaneous determination of these vitamins in 0.5 mi human serum or plasma. The vitamins were extracted from serum by methanol/iso-propanol (80/20, v/v) and n-hexane. The n-hexane phase was evaporated and injected to a reversed-phase (C-18) high-performance liquid chromatography system. Elution was performed with methanol/water (85:15, v/v) for 25-hydroxyvitamin D and retinol, and after that by methanol/water (98:2, v/v) for alpha-tocopherol. The eluate was monitored by a UV detector at 265 nm for detection of the vitamins. Baseline separation was obtained for all vitamins, and the system also permitted separate determinations of the D-2 and D-3 forms of 25-hydroxyvitamin D. The limit of detection and interassay variation for determination in 0.5 ml serum were 6.0 nmol/L and 6.2% for 25-hydroxyvitamin D-2, 6 nmol/l and 6.1% for 25-hydroxyvitamin D-3, 0.035 mu mol/L and 5.0% for retinol, and 1.2 mu mol/l and 5.5% for alpha-tocopherol.
引用
收藏
页码:339 / 343
页数:5
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