高效液相色谱法测定大鼠血浆中雷替曲塞浓度

HPLC Determination of Raltitrexed in Rat Plasma

目的:建立测定大鼠血浆中雷替曲塞的高效液相色谱方法。方法:血浆经甲醇沉淀蛋白后离心,上清液直接进样,HPLC- 紫外检测。色谱条件:Phenomenex Prodigy 5μODS3 100A色谱柱(5μm,0.46cm×25 cm)和Shim-pack GVP-ODS预柱。流动相为1%醋酸-甲醇-乙腈(60:25:15),流速1 mL·min-1。检测波长349 nm。结果:本法线性范围为25- 5000 ng·mL-1,最低定量限为25 ng·mL-1。血浆中雷替曲塞的绝对回收率为95.1%-105.8%,方法回收率为102.3%- 107.7%,日内RSD≤9.0%,日间RSD≤8.3%。结论:本方法适用于雷替曲塞在大鼠体内的药代动力学研究。

Objective:To develop an HPLC method for the determination of raltitrexed in rat plasma. Methods: Protein was precipitated by adding methanol into plasma, after centrifugation the supernatant was analysed by HPLC with a UV detector at 349 nm. A Phenomenex Prodigy 5μODS3 100A column(5 μm,0. 46 cm×25 cm) and a Shim - pack GVP - ODS guard column were used. The mobile phase consisted of 1 % acetic acid, methanol and acetonitrile in the proportion of 60: 25= 15(v/v) and the flow rate was 1 mL·min-1. Results:The linear range of the calibration curve for raltitrexed was 25-5000 ng·mL-1, and the limit of quantitation was about 25 ng·mL-1. The absolute recoveries of raltitrexed were 95.1%-105.8% , the analytical recoveries of raltitrexed were 102.3%-107.7%. Within-day RSD and between - day RSD were less than 9.0%and 8.3%.respectively. Conclusion: The method is confirmed to be appropriate for the pharmacokinetic study of raltitrexed in rats.