平衡透析结合LC-MS/MS法测定大鼠血浆中游离SN-38浓度及其药代动力学应用  

Determination of free SN-38 concentration in rat plasma by equilibrium dialysis combined with LC-MS/MS and its pharmacokinetic application

作　　者：罗珍华 宋成慧 李小川 张枢 张维维 李玉杰 段小群[3] 徐笑天 LUO Zhen-hua;SONG Cheng-hui;LI Xiao-chuan;ZHANG Shu;ZHANG Wei-wei;LI Yu-jie;DUAN Xiao-qun;XU Xiao-tian(School of Pharmacy,Guilin Medical University,Guilin Guangxi 541199;Xuhui Analysis Co.,Ltd.,Suzhou Jiangsu 215316;School of Biomedical Industry,Guilin Medical University,Guilin Guangxi 541199)

机构地区：[1]桂林医学院药学院,广西桂林541199 [2]苏州旭辉检测有限公司,江苏苏州215316 [3]桂林医学院生物医药产业学院,广西桂林541199

出　　处：《中南药学》2024年第1期71-77,共7页Central South Pharmacy

基　　金：广西科技重大专项(No.桂科AA22096026)。

摘　　要：目的 建立大鼠血浆中游离SN-38浓度的LC-MS/MS检测方法。方法 采用平衡透析法模拟SN-38在人体内与血浆蛋白结合的过程,在透析完成后分离出游离SN-38,以[2H5]SN-38为内标,用蛋白沉淀法预处理透析液侧(F侧)样品,取上清液进行LC-MS/MS分析;总SN-38及透析后血浆侧(T侧)SN-38样品的预处理方法与游离SN-38相同;分别考察两种方法的专属性、线性关系、精密度、准确度、提取回收率、基质效应、稳定性。结果F侧SN-38在0.04~80 ng·mL^(-1)内与峰面积线性关系良好(r> 0.99);提取回收率为98.6%~101.6%,基质效应为96.2%~97.2%;日内、日间RSD均<15%,且分别在不同条件下稳定性良好。大鼠血浆样品中的总SN-38在0.5~1000 ng·mL^(-1)内与峰面积线性关系良好(r> 0.99);提取回收率为97.3%~99.5%,基质效应为100.5%~101.2%;日内、日间RSD均<15%,且分别在不同条件下稳定性良好。结论 该方法简单易行、准确、灵敏、专属性好,适用于大鼠血浆中游离SN-38浓度的测定。Objective To establish an LC-MS/MS method to determine free SN-38 concentration.Methods Equilibrium dialysis was used to simulate the process of SN-38 binding with plasma protein,and the free SN-38 was isolated after the dialysis.The concentration of free SN-38 in dialysate side(F side) was determined by protein precipitation with [2H5]SN-38 as the internal standard and the supernatant was used for LC-MS/MS analysis.The total SN-38 and the SN-38 in the plasma(T side) after the dialysis were prepared by the same method as free SN-38.The specificity,linearity,precision,accuracy,extraction recovery,matrix effect and stability of both methods were determined respectively.Results SN-38 on the F side was well separated with good linear relationship(r > 0.99) at 0.04 ~ 80 ng·mL^(-1).The extraction recovery was 98.6% ~ 101.6%,and the matrix effect was 96.2% ~ 97.2%.Both inter-day and intra-day RSDs were lower than 15%,showing good stability in different conditions.The total SN-38 in the rat plasma samples was well separated by this method,with good linearity at 0.5 ~ 1000 ng·mL^(-1)(r > 0.99).The extraction recovery were 97.3% ~ 99.5%,and the matrix effect was 100.5% ~ 101.2%.RSDs of both inter-day and intra-day were lower than 15%,and also had good stability in different conditions.Conclusion The method is simple,accurate,sensitive and specific,which is suitable for the determination of free SN-38 concentration in rat plasma.

关 键 词：SN-38 游离浓度 平衡透析法 LC-MS/MS 

分 类 号：R96[医药卫生—药理学]