HPLC-MS/MS法测定人血浆中氯法拉滨的浓度

目的:建立测定人血浆中氯法拉滨浓度的方法。方法:血浆样品经甲基叔丁基醚提取处理后,采用高效液相色谱串联质谱电喷雾(HPLC-MS/MS)法进行测定,色谱柱为Thermo C18,流动相为乙腈-水(250∶3,其中含4 mmol·L-1乙酸铵和0.3%甲酸),流速为0.5 mL·min-1,进样量为10μL,内标为克拉屈滨。用于定量氯法拉滨和克拉屈滨的离子反应分别为m/z 304.2→170.0、m/z286.1→170.0。每个样品的分析时间为5 min。结果:氯法拉滨和克拉屈滨的保留时间分别为3.83、3.90 min。氯法拉滨血药浓度在2.5～500 ng·mL-1范围内线性关系良好(r=0.999 9),日内、日间RSD均<14%,方法回收率为95.64%～99.23%。结论:本方法简便、灵敏、快速、准确,适用于氯法拉滨的血药浓度测定及药动学研究。

Determination of the Concentration of Clofarabine in Human Plasma by HPLC-MS/MS

OBJECTIVE： To establish a method for the concentration determination of clofarabine in human plasma. METHODS： The plasma samples were extracted with methyl tert-butyl ether, then was determined by HPLC-MS/MS. Thermo C18 column was used as analytic column with mobile phase consisted of acetonitrile-water （250 ∶ 3, containing 4 mmol·L-1 ammonium acetate and 0.3% formic acid） at flow rate of 0.5 mL·min-1. The injection volume was 10 μL and cladribine was used as the internal standard （IS）. Clofarabine and cladribine were detected on multiple reaction monitoring （MRM） by the transitions from the precursor to the product ion （m/z 304.2→170.0 and m/z 286.1→170.0）. And the total running time was 5 min. RESULTS： The retention time of clofarabine and cladribine were 3.83 min and 3.90 min. The linear range of clofarabine was 2.5～500 ng·mL-1（r=0.999 9）. The RSD of intra-day and inter-day were both less than 14% and the average recovery was 95.64%～99.23%. CONCLUSION： Established method is simple, sensitive, rapid and accurate, and it is suitable for plasma concentration determination and pharmacokinetics study of clofarabine.