1986 年 35 巻 3 号 p. 329-332
A high performance liquid chromatographic method is described for the determination of antipain which is a protease inhibitor and also a possible therapeutic drug for muscular destrophy. Antipain was converted to the fluorescent derivative by reaction with benzoin, a fluorogenic reagent for guanidino group. The derivative was then determined on a reversed phase column with isocratic elution within 15 min by using fluorescence detector. The detection limit of antipain in mouse serum was 0.42μg/ml, which corresponds to the amount of 240 pg (400 fmol) in an injection volume of 100μ1 at a signal to noise ratio of 2. The calibration graph for the compound was linear up to at least a concentration of 30 μg/ml in the serum. The proposed method was simple, rapid and sensitive enough to permit the quantification of antipain in as little as 40 μl of mouse serum dosed with the compound.