Determination and pharmacokinetic study of norcantharidin in human serum by HPLC-MS/MS method

A sensitive, simple and selective high-performance liquid chromatography–tandem mass spectrometry method was developed and applied to the determination of norcantharidin concentration in human serum. Norcantharidin (NCTD) and cyclophosphamide (IS) in serum were extracted with acetone, separated on a C18 reversed-phase column, gradiently eluted with a mobile phase of acetonitrile–water containing 2 mm ammonium acetate and 0.1% formic acid (pH 3), ionized by positive ion pneumatically assisted electrospray and detected in the multi-reaction monitoring mode using precursor product ions of m/z 169.3 123.1 for NCTD and 261.2 140.2 for IS, respectively. The linear range of the calibration curve for NCTD was 2.5–50 ng/mL, with a lowest limit of quantification of 2.5 ng/mL, and the intra/inter-day RSD was less than 10%. The method was suitable for determination of low NCTD concentration in human serum after therapeutic oral doses, and has been successfully used for pharmacokinetic studies in healthy Chinese volunteers. Copyright © 2007 John Wiley & Sons, Ltd.