Fluorescence High Performance Liquid Chromatography of Bile Acid 3-Sulfates Using l-Bromoacetylpyrene as Prelabeling Reagent (生体成分の高速液体クロマトグラフィ-)

概要

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A method for the simultaneous determination of bile acid 3sulfates in human serum without prior hydrolysis or solvolysis is developed. Free and glycine conjugated bile acid 3-sulfates were labeled with 1-bromoacetylpyrene in acetonitrile by using cyclohexyl-18-crown-6-ether as catalyst. Derivatized bile acid 3-sulfates were separated stepwisely on a reversed phase column by using acetonitrile-0.01 M sodium acetate-methanol (80 : 100 : 35, v/v) and acetonitrile-water-methanol (50 : 100 : 30, v/v) as mobile phase at a flow rate of 1.5 ml/min. The effluent was monitored by a fluorophotometer at 370 run (excitation) and 440 nm (emission). Linearities of fluorescence intensities (peak heights) with the amounts of each bile acid 3-sulfates were obtained in the range from 25 to 125 pmol. The present method has proved to be applicable to the determination of bile acid 3-sulfates in serum, bile and urine.
社団法人日本分析化学会の論文
1984-07-05