RP-HPLC Method for the Estimation of 6-Mercaptopurine in spiked human plasma and pharmaceutical formulations

A simple, rapid and accurate reverse-phase high-performance liquid chromatography (RP-HPLC) method for the determination of 6-mercaptopurine (6-MP) in pharmaceutical formulations and spiked human plasma samples has been developed and validated. The assay of the drug was performed on a CLC C18 (5 μm, 25 cm × 4.6 mm i.d.) with UV detection at 325 nm. The mobile phase consisted of methanol-water mixture in the ratio of 90: 10, and a flow rate of 1 mL/min was maintained. The standard curve was linear over the range of 25–150 μg/mL (r2 = 0.9964). Analytical parameters have been evaluated. Between and within-day precision and accuracy were acceptable down to the limit of quantification of 9.5 μg/mL in plasma. The proposed method was validated for accuracy and precision. Statistical analysis proves that the method was found to be simple, precise, accurate, rapid and reproducible and can be used for the routine determination of 6-mercaptopurine. The proposed method was successfully applied to the determination of 6-mercaptopurine in spiked human plasma and pharmaceutical formulations. The method will be useful for routine quality control analysis.