High-performance liquid chromatographic method for quantification of busulfan in plasma after derivatization by tetrafluorothiophenol.

M H Quernin, B Poonkuzhali, Y Médard, D Dennison, A Srivastava, R Krishnamoorthy, M Chandy, E Jacqz-Aigrain

Index: J. Chromatogr. B. Biomed. Sci. Appl. 721(1) , 147-52, (1999)

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Abstract

A high-performance liquid chromatographic (HPLC) method was developed and validated for the determination of busulfan in plasma. Busulfan was extracted in toluene, derivatized by 2,3,5,6-tetrafluorothiophenol to obtain di-TFTP-butane, the derivatization product was then re-extracted in toluene and injected into the HPLC system with ultraviolet detection (wavelength: 275 nm). Recovery from extraction was 80%, the limit of quantification was 50 ng/ml and linearity ranged from 50 to 2000 ng/ml. In addition, forty-two samples obtained from pediatric patients treated with busulfan were analyzed by the HPLC and GC-MS assays based on the same derivatization procedure. The correlation between the di-TFTP-butane concentrations was highly significant (p<0.0001), demonstrating that the two methods were in good agreement.