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,1.9μm),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1～1 000μg/L范围内线性关系良好(r=0.997 8,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0.2μg/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。
An ultra-high performance liquid chromatography-tandem mass spectrometry （UH- PLC-MS/MS） method for the determination of mitomycin C （MMC） in rabbit plasma was estab- lished. 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 comple- ted 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 1 000 μg/L （ r =0. 997 8, 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 stand- ard 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.
出处
《色谱》
CAS
CSCD
北大核心
2012年第2期154-159,共6页
Chinese Journal of Chromatography
基金
国家"重大新药创制"科技重大专项(No.2008ZX09305-002)
全军"十一五"计划项目(No.06MA013)
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
ultra-high performance liquid chromatography-tandem mass （UHPLC-MS/MS）
electrospray ionization （ESI）
mitomycin C
New Zealand big plasma spectrometry white rabbit
