液-质联用法测定人血浆中利伐沙班的浓度

Determination of rivaroxaban in human plasma by HPLC-MS/MS

目的：建立液-质联用测定人血浆中利伐沙班浓度的方法。方法：采用Thermo Hypurity C18色谱柱（150 mm×2.1mm,5μm）;流动相：5 mmol·L-1醋酸铵-乙腈（70∶30）;流速：0.3 ml·min-1;柱温：30℃;MRM模式检测（离子对436.1/144.9）,内标法定量。结果：血浆利伐沙班浓度在0.6-300 ng·ml-1范围内线性关系良好（r=0.999 7）。低浓度点RSD小于20%,其余浓度点RSD均小于15%。血浆样品冻存（-20℃）30 d稳定性良好,反复冻融3次及提取后室温放置6 h条件下,样品浓度均无显著变化。结论：所建方法快速简便、灵敏准确,本法可用于利伐沙班血浆中浓度的测定以及药动学研究。

OBJECTIVE To establish a HPLC MS/MS method for the determination of rivaroxaban in human plasma. METHODS The analysis was carried out by using Thermo Hypurity C18 column（150 mm×2. 1 mm,5μm）. The mobile phase was consisted of 5 mmol, L-1 ammonium acetate-acetonitrile （70：30） with a flow rate at 0. 3 ml,min-1. The column temperature was 30 ℃ and the concentrations in plasma were determined at ion pairs （436. 1/144.9） by MRM of HPLC-MS/MS.RESULTS The linear range of pseudoephedrine hydrochloride was 0. 6 - 300 ng. ml-1 （r = 0. 999 7）. Both the inter- and intra- precisions were less than 20% at the low concentration and less than 15% at the middle and high concentrations. The spiked plasma samples were stable at - 20 ℃ in a long-term stability test. After three freeze-thaw cycles of the spiked samples and 6h after extraction, the changes in levels of rivaroxaban were not obvious. CONCLUSION The method can be used for the deter- mination of rivaroxaban in human plasma and the study of its pharmacokinetics.