色谱

色谱 ›› 2012, Vol. 30 ›› Issue (02): 154-159.DOI: 10.3724/SP.J.1123.2011.10034

• 研究论文 • 上一篇    下一篇

超高效液相色谱-串联质谱法测定兔血浆中的丝裂霉素C

汤瑶1, 张双庆1, 李响2, 孙旭1, 闻镍1, 于敏1, 彭莉莉3, 李进让3, 李佐刚1*, 李波1   

  1. 1. 中国食品药品检定研究院 国家药物安全评价监测中心, 北京 100176; 2. 吉林大学生命科学学院, 长春 130012; 3. 中国人民解放军海军总医院, 北京 100048
  • 收稿日期:2011-10-28 修回日期:2011-12-07 出版日期:2012-02-28 发布日期:2012-03-22
  • 通讯作者: 李佐刚,研究员,主要从事临床前药动学研究和GLP管理. Tel: (010)67872233-8312,
  • 基金资助:

    国家“重大新药创制”科技重大专项(No. 2008ZX09305-002)和全军“十一五”计划项目(No. 06MA013)

Determination of mitomycin C in rabbit plasma by ultra-high performance liquid chromatography-tandem mass spectrometry

TANG Yao1, ZHANG Shuangqing1, LI Xiang2, SUN Xu1, WEN Nie1, YU Min1, PENG Lili3, LI Jinrang3, LI Zuogang1*, LI Bo1   

  1. 1. National Center for Safety Evaluation of Drugs, National Institutes for Food and Drug Control, Beijing 100176, China; 2. College of Life Sciences, Jilin University, Changchun 130012, China; 3. Navy General Hospital, the Chinese People’s Liberation Army, Beijing 100048, China
  • Received:2011-10-28 Revised:2011-12-07 Online:2012-02-28 Published:2012-03-22

PDF

406

被引次数

20

摘要： 建立了采用超高效液相色谱-串联质谱测定兔血浆中丝裂霉素C的方法。以兔空白血浆为基质,通过添加标准溶液的方法配制含丝裂霉素C和内标物曲安奈德的样品,选用乙酸乙酯为提取溶剂,液-液萃取法处理血浆样品。采用Hypersil Gold C18分析柱(50 mm×2.1 mm, 1.9 μm),流动相为甲醇-0.1%甲酸水溶液(90:10, v/v),等度洗脱,流速0.2 mL/min,柱温35 ℃,在3 min内实现了快速分离。采用电喷雾正离子(ESI+)模式电离,选择反应监测(SRM)模式检测,以曲安奈德作为内标物进行定量。用于监测的定量离子对分别为丝裂霉素C m/z 335.2→242.2和曲安奈德m/z 435.2→397.3/415.2,用基质匹配标准溶液法进行定量。结果表明: 兔血浆中丝裂霉素C的质量浓度在1～1000 μg/L范围内线性关系良好(r=0.9978,权重系数(weighting): 1/x2);血浆中丝裂霉素C的检出限(信噪比为3)为0.2 μg/L;其平均回收率为85%～ 115%;日内及日间的相对标准偏差(RSDs)均小于15%,满足生物样品检测的要求。该方法可用于兔气管外壁给药后的血浆样品中丝裂霉素C的检测。本方法选择性强、灵敏度高、操作简便快速、重现性好,适用于丝裂霉素C药代动力学等方面的研究。

关键词: 超高效液相色谱-串联质谱法, 电喷雾离子化, 丝裂霉素C, 新西兰大白兔, 血浆

Abstract: An ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for the determination of mitomycin C (MMC) in rabbit plasma was established. The blank rabbit plasma sample solutions added with mitomycin C and triamcinolone acetonide (internal standard, IS) were prepared. The solutions containing MMC and IS were extracted from the plasma with ethyl acetate using liquid-liquid extraction method. A Hypersil Gold C18 column (50 mm×2.1 mm, 1.9 μm) was employed and the column temperature was set at 35 ℃. The isocratic elution of methanol and 0.1% (v/v) formic acid aqueous solution as mobile phase was performed at a flow rate of 0.2 mL/min, and a rapid separation was completed within 3 min. The electrospray was operated in the positive ionization mode and the MMC and IS were identified in selected reaction monitoring (SRM) mode. The monitoring ions of MMC and IS were m/z 335.2→242.2 and m/z 435.2→397.3/415.2, respectively, which were used to qualify and quantify the targets by the method of matrix-matched standard solution. The calibration curve showed good linearity within the mass concentrations of 1 to 1000 μg/L (r=0.9978, weighting: 1/x2). The limit of detection (S/N=3) was 0.2 μg/L. The recoveries were from 85% to 115% at the spiked levels of 1, 5, 100 and 800 μg/L, and the relative standard deviations (RSDs) of intra- and inter-day were both less than 15%. The method can meet the determination requirements of biological samples, and can be used for the determination of mitomycin C in rabbit plasma after the administration of mitomycin C. The method is selective, sensitive, convenient, rapid and reproducible in the determination of mitomycin C, and also can be used for the pharmacokinetics research of mitomycin C in plasma.

Key words: electrospray ionization (ESI), mitomycin C, New Zealand big white rabbit, plasma, ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS)