LC-MS测定大鼠脑脊液与血浆中西达本胺浓度及其药动学研究

Pharmacokinetic Study of Chidamide Concentration in Cerebrospinal Fluid and Plasma of Rats by LC-MS

目的建立快速检测SD大鼠脑脊液和血浆中西达本胺浓度的LC-MS测定方法,并应用于大鼠静脉注射西达本胺后的脑脊液和血浆中药动学研究。方法对建立的检测大鼠脑脊液和血浆中西达本胺浓度的LC-MS分析方法进行专属性、精密度、准确性、基质效应及稳定性等考察验证。♂SD大鼠12只,分为2组,每组6只,分别静脉注射给予西达本胺3,6 mg·kg^(-1)剂量,于给药后0.083,0.25,0.5,1,1.5,2,3 h采集脑脊液和血浆,采用LC-MS测定大鼠脑脊液和血浆中西达本胺的浓度变化,并利用DAS药动学软件拟合脑脊液和血浆中AUC、C_(max)、T_(max)、t_(1/2)等主要药动学参数,计算血脑屏障透过率。结果方法学研究显示,大鼠脑脊液中西达本胺线性范围为0.2~64 ng·mL^(-1)(r=0.999 0,定量下限为0.2 ng·mL^(-1)),血浆中西达本胺的线性范围为2~1 000 ng·mL^(-1)(r=0.999 1,定量下限为2 ng·mL^(-1)),脑脊液和血浆中西达本胺的日内精密度、日间精密度、准确度均符合方法学验证要求。大鼠药动学研究显示,大鼠静脉注射给予西达本胺3,6 mg·kg^(-1)剂量后,脑脊液中的AUC_(0-t)分别为16.01,32.09 ng^(-1)·mL^(-1)·h,C_(max)分别为17.58,28.37 ng·mL^(-1),t_(1/2)分别为1.14,1.23 h;血浆的AUC_(0-t)分别为2 570.11,4 537.63 ng^(-1)·mL^(-1)·h,C_(max)分别为4 366.54,9 103.97 ng·mL^(-1),t_(1/2)分别为0.83,0.77 h。2组西达本胺的平均血脑屏障透过率分别为0.62%和0.72%。结论本研究建立了简便、迅速、灵敏的检测大鼠脑脊液和血浆中西达本胺浓度的分析方法,并成功应用于大鼠药动学研究,静脉注射西达本胺能够有效地透过血脑屏障,可以实现直接作用于脑部肿瘤的目的。

OBJECTIVE To establish a rapid LC-MS method for the determination of chidamide in cerebrospinal fluid and plasma of SD rats,and apply it to the pharmacokinetic study of cerebrospinal fluid and plasma in rats after intravenous injection of chidamide.METHODS The specificity,precision,accuracy,matrix effect and stability of the established LC-MS method for detecting the concentration of chidamide in cerebrospinal fluid and plasma of rats were investigated and verified.Twelve male SD rats were divided into 2 groups with 6 rats in each group.They were given 3,6 mg·kg^(-1)intravenously,respectively.Cerebrospinal fluid and plasma were collected at 0.083,0.25,0.5,1,1.5,2,3 h after administration.The concentrations of chidamide in cerebrospinal fluid and plasma of rats were determined by LC-MS method,and the pharmacokinetic parameters of AUC,C_(max),T_(max)and t_(1/2)in cerebrospinal fluid and plasma were fitted by DAS pharmacokinetic software,and the blood-brain barrier permeability was calculated.RESULTS Methodological studies showed that the linear range of chidamide in rat cerebrospinal fluid was 0.2~64 ng·mL^(-1)(r=0.999 0,lower limit of quantification was 0.2 ng·mL^(-1)),and the linear range of chidamide in plasma was 2~1 000 ng·mL^(-1)(r=0.999 1,lower limit of quantification was 2 ng·mL^(-1)).Inter-day and intra-day variability and accuracy of chidamide in cerebrospinal fluid and plasma all met the requirements of methodological verification.Pharmacokinetic study of rats showed that the AUC_(0-t)in cerebrospinal fluid was 16.01,32.09 ng^(-1)·mL^(-1)·h after intravenous administration of 3,6 mg·kg^(-1),C_(max) were 17.58,28.37 ng·mL^(-1),t_(1/2)were 1.14,1.23 h.The AUC,C_(max) of plasma were 2 570.11,4 537.63 ng^(-1)·mL^(-1)h,4 366.54,9 103.97 ng mL^(-1),t_(1/2)were 0.83,0.77 h,respectively.The average blood-brain barrier permeability of chidamide in the two groups was 0.62% and 0.72%,respectively.CONCLUSION The study establishes a simple,rapid and sensitive method for the determination of chidamide concentration in cerebrospinal fluid and plasma of rats,and successfully applies to the pharmacokinetic study of rats.Intravenous administration of chidamide can effectively cross the blood-brain barrier and directly act on brain tumors.