Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 501 Haike Road, Shanghai 201203, PR China; Department of Clinical Pharmacy, Bethune International Peace Hospital, 398 West Zhongshan Road, Shijiazhuang 050082, PR China.
Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 501 Haike Road, Shanghai 201203, PR China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2014 Mar 1;951-952:62-8. doi: 10.1016/j.jchromb.2014.01.017. Epub 2014 Jan 19.
Sinotecan is a camptothecin analog, currently under clinical testing as an antitumor medication. We developed and validated a rapid, specific and reliable liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for simultaneous quantification of sinotecan and its active metabolite, 7-hydroxyethyl-camptothecin (7-HEC), in human blood. Aliquots (200μL) of heparinized blood samples were processed by deproteinization with 400μL acetonitrile each. Chromatographic analyte separation used an Agilent Zorbax SB C8 column (4.6mm×150mm, 5μm) and methanol/10mM ammonium acetate/formic acid (70/30/0.14, v/v/v) as mobile phase, at a flow rate of 0.60mL/min. A Thermo Finnigan TSQ Quantum Ultra tandem mass spectrometer was operated in multiple-reaction monitoring mode. The precursor-to-product ion transitions m/z 493→m/z (331+375) for sinotecan, m/z 393→m/z (233+261) for 7-HEC, and m/z 396→m/z 352 for d3-SN38 (IS) were used for quantification. The method was validated for 1.0-500ng/mL for sinotecan and 0.5-250ng/mL for 7-HEC using 200μL of blood sample. Total time for each chromatograph was ∼6.0min. The intra- and inter-day precision and accuracy of the quality control samples at low, medium, and high concentration levels exhibited relative standard deviations (RSD)<13.8% and the accuracy values ranged from -5.3% to 2.4%. The method was successfully applied to a pharmacokinetic study of sinotecan in cancer patients.
喜树碱是一种喜树碱类似物，目前正在临床测试中作为抗肿瘤药物。我们开发并验证了一种快速、特异和可靠的液相色谱-串联质谱（LC-MS/MS）方法，用于同时定量人血液中的喜树碱及其活性代谢物 7-乙基喜树碱（7-HEC）。用 400μL 乙腈沉淀处理肝素化血样等分试样（200μL）。用 Agilent Zorbax SB C8 柱（4.6mm×150mm，5μm）和甲醇/10mM 乙酸铵/甲酸（70/30/0.14，v/v/v）作为流动相进行色谱分析物分离，流速为 0.60mL/min。Thermo Finnigan TSQ Quantum Ultra 串联质谱仪以多反应监测模式操作。用于定量的前体到产物离子跃迁 m/z 493→m/z（331+375）为喜树碱，m/z 393→m/z（233+261）为 7-HEC，m/z 396→m/z 352 为 d3-SN38（IS）。使用 200μL 血样，喜树碱的定量下限为 1.0-500ng/mL，7-HEC 的定量下限为 0.5-250ng/mL。每个色谱图的总时间约为 6.0min。低、中、高浓度质控样品的日内和日间精密度和准确度的相对标准偏差（RSD）<13.8%，准确度值范围为-5.3%至 2.4%。该方法成功应用于癌症患者喜树碱的药代动力学研究。
