High-performance liquid chromatographic method for monitoring leupeptin in mouse serum and muscle by pre-column fluorescence derivatization with benzoin.

A high-performance liquid chromatographic method is described for the determination of leupeptin, a possible therapeutic drug for muscular dystrophy, in mouse serum and muscle. Leupeptin is reduced with sodium borohydride to leupeptinol, and then converted to a fluorescent derivative with benzoin. The derivative is separated on a reversed-phase column (LiChrosorb RP-18) with isocratic elution and determined with fluorescence detection. The detection limits of leupeptin in serum and muscle are 250 pmol/ml (107 ng/ml) and 500 pmol/g (214 ng/g), respectively, corresponding to approximately 150 fmol each in a 100-microliters injection volume. This method is simple and sensitive enough to permit the quantification of leupeptin in biological samples from mice dosed with leupeptin.