METHOD FOR MEASUREMENT OF L-PHENYLALANINE MUSTARD IN MOUSE AND DOG BY HIGH-PRESSURE LIQUID-CHROMATOGRAPHY

Abstract

The distribution of L-phenylalanine mustard (L-PAM [an antineoplastic agent]) was studied in dogs and mice by high-pressure liquid chromatography. Separation of L-PAM from its products of hydrolysis was accomplished with a .mu.-Bondapak C18 column, a solvent system composed of 2-methyoxethanol/0.1% acetic acid, and solvent programming with a step gradient. Complete separation was effected in less than 15 min. The half-life for disappearance of L-PAM from mouse blood was 41 min; that from dog blood was 29 min. The monohydroxy derivative of L-PAM, L-MOH [4-[(2-chloroethyl)(2-hydroxyethyl)amino]-L-phenylalanine] disappeared from dog serum with a half-life of 32 min. L-MOH was not detectable in mouse tissue other than blood at times greater than 15 min after injection. In the dog at 4 h after injection, the tissue/serum concentration ratios were > 1 for liver, spleen, intestine, skeletal muscle, urinary bladder and gallbladder. The concentration of L-PAM in the bile was approximately 500 times higher than that in serum.