High-Performance Liquid Chromatographic Measurement of Cyclophosphamide in Serum

The analysis of cyclophosphamide [N,N-bis(2-chlorethyl)-tetrahydro-2H-1,3,2-oxazaphosphorin-2-amine -2-oxide] by high-performance liquid chromatography using ultraviolet detection is described. The method will enable measurement of serum concentrations of cyclophosphamide over a period of approximately 24 h after a dose of 150 mg, and requires 1 ml of serum. The between-day precision of the assay at concentrations of 0.3, 1.0, and 15.0 mg/L generated coefficients of variation of 11.8, 12.2, and 7.7%, respectively. Percentages of analytical recovery of cyclophosphamide and internal standard (5-ethyl-5-p-tolylbarbituric acid) were 63 and 73%, respectively. Preliminary data providing the half-life for two patients with normal renal function are presented.