共役ジエンを標的とする新蛍光標識試薬の開発ならびに同試薬のビタミンDおよびA代謝物定量への応用

概要

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A new sensitive and highly reactive fluorescence-labeling reagent (DMEQ-TAD) targeting conjugated dienes was developed. The fluorescent dienophile (DMEQ-TAD), in which fluorescent dimethoxyquinoxalinone (DMEQ) group is substituted via ethylene spacer on a dienophile, 1,2,4-triazoline-3,5-dione (TAD), was synthesized in 8 steps in 24% overall yield from dinitroveratrole. The reactions of DMEQ-TAD with six major vitamin D metabolites and some synthetic analogs were examined under various conditions. The reaction produced the corresponding 6,19-cycloadduct as a pair of the C (6) epimers (8 and 9) in quantitative yield. The structures of the adducts (8 and 9) including the stereochemistry at C (6) were unambiguously determined. The fluorescence-labeled vitamins were analyzed by HPLC with fluorescence detector. The new fluorometric method was used in the assay of plasma 25-OH-D_3, 24,25 (OH)_2D_3, and 25,26 (OH)_2D_3. The method was proved to be precise and reliable by comparing with the HPLC-UV method. The reaction of DMEQ-TAD with vitamin A metabolites and fluorometric assay of plasma retinoic acid was also examined.
日本ビタミン学会の論文
1994-01-25