Liquid chromatographic analysis, stability and protein binding studies of the anti-HIV agent benzoic acid, 2-chloro-5[[(1-methylethoxy)thioxomethyl]amino]-,1-methylethyl ester.

A method was developed for the assay of benzoic acid, 2-chloro-5-[[(1-methylethoxyl)thioxomethyl]amino]-,1-meth yle thyl ester (NSC 629243) in hamster, mouse, human and, to a limited extent, dog plasma. Protein in 0.5 ml of plasma was precipitated with four volumes of methanol and the supernatant was analysed for NSC 629243 by LC. Liquid chromatography was carried out on a reversed-phase Nova-Pak C18 column, with a mobile phase of 60% acetonitrile in water at 1 ml min-1, and the compound was quantified with a UV detector set at 283 nm. Two standard curves of NSC 629243 were needed to cover a concentration range of 0.05-100 micrograms ml-1. All standard curves had correlation coefficients > 0.999. In practice, the minimum quantifiable concentration was approximately 0.05 microgram ml-1 in 0.5 ml of plasma. NSC 629243 appeared to have good stability at 37 degrees C in hamster, human and dog plasma at concentrations of 1 and 50 micrograms ml-1 (at least 80% remained in plasma after a 4 h incubation). Breakdown occurred in mouse plasma after 1 h at 37 degrees C, with extensive breakdown occurring after 24 h. NSC 629243 was extensively bound to plasma proteins of Syrian hamsters and humans. The extent of binding ranged from a minimum of 88.6% to a maximum of 99.9% over a concentration range of ca 1-100 micrograms ml-1.

Effect of dose on the percutaneous absorption of 2- and 4-chloronitrobenzene in rats.

The effect of dose on the dermal absorption of 2- and 4-chloronitrobenzene (2- and 4-CNB) has been investigated in rats following nonocclusive protective dermal application on an area of 4 cm2 per animal at approximately 0.0325, 0.325, and 3.25 mg/cm2 (0.65, 6.5, and 65 mg/kg, respectively). At the three-dose levels, 33-40% and 51-62% of the dose of 2- and 4-CNB, respectively, was absorbed from the skin within 72 hr. The balance of the dose was recovered in the protective device and the organic trap (i.e. that portion unavailable for dermal absorption). The absorbed radioactivity was excreted in urine (21-28% of dose, 2-CNB; 43-45%, 4-CNB) and feces (11-15%, 2-CNB; 5-12%, 4-CNB). The extent and rate of dermal absorption and urinary and fecal excretion of 2-CNB were linear over the 0.65-65 mg/kg dose range; for 4-CNB they were linear over the 0.65-6.5 mg/kg dose, and nonlinear at the 65 mg/kg dose.