DETERMINATION OF ALTRETAMINE IN HUMAN PLASMA WITH HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY

A fast, simple, and sensitive isocratic HPLC method has been developed and validated for the determination of the anticancer drug altretamin e in human plasma. Spiked serum samples and clinical plasma samples ar e extracted with acetonitrile at 4 degrees C and the precipitate remov ed by filtration. The plasma sample volume required (ca. 0.2 ml) is sm all and the total analysis time is less than 15 min per sample (includ ing batch-wise pre-treatment). Recovery of altretamine is 99 to 106% f or pooled human serum spiked with altretamine in the range 200 ng/ml t o 10 mg/ml. In this concentration range, the R.S.D. varies from 1 to 8 %. The limit of quantitation is ca. 150 ng/ml for an R.S.D. of 10%. Th e intra-day R.S.D. for human samples spiked at 5 mg/ml varied between 1.7 and 4%; the inter-day R.S.D. at this concentration was ca. 3%. A p reliminary study with one patient receiving 260 mg/m2 by mouth indicat ed that the peak altretamine concentration was significantly lower aft er a standard breakfast than in the fasting state.