Wei Chun-min, Zhang Rui, Wang Ben-jie, Yuan Gui-yan, Guo Rui-chen
Institute of Clinical Pharmacology, Qilu Hospital of Shandong University, Jinan 250012, People's Republic of China.
Biomed Chromatogr. 2008 Jan;22(1):44-9. doi: 10.1002/bmc.892.
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.
建立了一种灵敏、简便且选择性高的高效液相色谱-串联质谱法，并将其应用于测定人血清中去甲斑蝥素的浓度。血清中的去甲斑蝥素（NCTD）和环磷酰胺（内标，IS）用丙酮萃取，在C18反相柱上分离，用含2 mM醋酸铵和0.1%甲酸（pH 3）的乙腈-水流动相进行梯度洗脱，通过正离子气动辅助电喷雾进行离子化，并在多反应监测模式下检测，NCTD的前体离子→产物离子分别为m/z 169.3→123.1，IS的为m/z 261.2→140.2。NCTD校准曲线的线性范围为2.5 - 50 ng/mL，最低定量限为2.5 ng/mL，日内/日间相对标准偏差小于10%。该方法适用于测定治疗口服剂量后人血清中低浓度的NCTD，已成功用于健康中国志愿者的药代动力学研究。
