血浆中25-羟基维生素D3和24R,25-二羟基维生素D3的荧光测定法。
Fluorometric assay of 25-hydroxyvitamin D3 and 24R,25-dihydroxyvitamin D3 in plasma.
作者信息
Shimizu M, Gao Y, Aso T, Nakatsu K, Yamada S
机构信息
Institute for Medical and Dental Engineering, Tokyo Medical and Dental University, Japan.
出版信息
Anal Biochem. 1992 Aug 1;204(2):258-64. doi: 10.1016/0003-2697(92)90236-z.
The first practical fluorometric assay of plasma 25-hydroxyvitamin D3 (25-OH-D3) and 24R,25-dihydroxyvitamin D3 (24,25-(OH)2D3) is described. The method uses a highly fluorescent dienophile, 4-[2-(6,7-dimethoxy-4-methyl-3-oxo-3,4-dihydroquinoxalyl)ethyl]-1, 2,4- triazoline-3,5-dione (DMEQ-TAD), to fluorescence-label vitamin D. Vitamin D metabolites were roughly purified with a short cartridge column followed by HPLC, labeled with DMEQ-TAD, and the product was analyzed on HPLC. In the assay of 25-OH-D3 the new fluorometric method was compared with the HPLC-uv method and was confirmed to be as accurate and reliable (CV, 4-5%) as the HPLC-uv method. Plasma 24,25-(OH)2D3 was accurately assayed by the HPLC-FL method, where the standard addition method was successfully used to calculate the overall recovery.
本文描述了血浆25-羟基维生素D3（25-OH-D3）和24R,25-二羟基维生素D3（24,25-(OH)2D3）的首个实用荧光测定法。该方法使用一种高荧光亲双烯体4-[2-(6,7-二甲氧基-4-甲基-3-氧代-3,4-二氢喹喔啉基)乙基]-1,2,4-三唑啉-3,5-二酮（DMEQ-TAD）对维生素D进行荧光标记。维生素D代谢物先用短柱进行初步纯化，然后通过高效液相色谱（HPLC），用DMEQ-TAD进行标记，产物再通过HPLC进行分析。在25-OH-D3的测定中，将新的荧光测定法与HPLC-紫外检测法进行了比较，结果证实其准确性和可靠性与HPLC-紫外检测法相当（变异系数为4-5%）。通过HPLC-荧光检测法可准确测定血浆中的24,25-(OH)2D3，其中成功采用标准加入法计算总回收率。
Zotero 插件