摘要
目的:建立一种简便、快速的LC-MS/MS法测定大鼠脑组织中奥拉西坦的浓度,并对SD大鼠静脉注射奥拉西坦后的脑组织分布进行研究。方法:SD大鼠脑组织样品用0.9%氯化钠溶液匀浆后,以吡拉西坦为内标,用乙腈沉淀蛋白,采用LC-MS/MS系统进行分离和测定。色谱柱为Agilent Poroshell 120 SB-C_(18)柱(100 mm×4.6 mm,2.7μm),流动相为30%甲醇-70%水(含10 mmol·L^(-1)甲酸铵,0.1%甲酸),流速为0.2 mL·min^(-1),柱温为30℃。质谱条件:电喷雾离子源(ESI),正离子模式,多反应离子监测(MRM),检测离子为奥拉西坦m/z 158.6→113.6,内标吡拉西坦m/z 142.6→125.4。SD大鼠分别单次静脉注射奥拉西坦500、1 000、2 000 mg·kg^(-1),于不同时间点分别采集脑组织,测定脑组织中奥拉西坦的浓度。结果:SD大鼠脑组织匀浆液质量浓度在1.0~100.0μg·mL^(-1)范围内线性关系良好,定量下限为1.0μg·mL^(-1),批内和批间精密度RSD均小于5.7%,准确度为96.3%~105.2%。大鼠分别单次静脉注射500、1 000、2 000 mg·kg^(-1) 3个剂量的奥拉西坦后,主要药动学参数t1/2分别为(3.56±0.27)h、(3.36±0.49)h、(3.60±0.79)h,C_(max)分别为(24.78±16.06)μg·g^(-1)、(50.79±25.92)μg·g^(-1)、(82.21±19.33)μg·g^(-1),MRT0-t分别为(2.54±0.02)h、(2.38±0.30)h、(2.57±0.08)h,AUC0-t分别为(48.59±10.92)μg·h·g^(-1)、(94.54±9.30)μg·h·g^(-1)、(201.35±18.87)μg·h·g^(-1)。结论:本测定方法可用于大鼠奥拉西坦药代动力学在脑组织中的分布研究;大鼠分别单次静脉注射3个剂量的奥拉西坦后,奥拉西坦进入脑组织的时间较快,浓度较高,持续时间较长,利于发挥活化和改善脑组织功能的作用。
Objective:To develop a simple and rapid HPLC-MS/MS method for the determination of oxiracetam concentration in rat brain tissue,and to study the distribution of oxiracetam in the brain tissues after being injected intravenously in SD rats.Methods:The tissue samples of SD rat brain were firstly homogenized with 0.9% sodiumchloride solution.Using piracetam as an internal standard(IS)and acetonitrile as a protein precipitant,the samples were separated and determined by LC-MS/MS system.Agilent Poroshell 120 SB-C_(18) analytical column(100 mm×4.6 mm 2.7 μm)was used in the system,and the mobile phase was prepared with methanol and water in a volume ratio of 30∶70,containing 10 mmol·L-1 ammonium acetate and 0.1% formic acid.The flow rate was 0.2 mL·min-1 and the column temperature was set at 30 ℃.The parameters for MS were as follows:electrospray ionization source(ESI)was operated in positive ion mode;the quantification was performed using multiple reaction monitoring(MRM)mode to monitor the precursor-to-product ion transitions of m/z 158.6 → 113.6 for oxiracetam and m/z 142.6 → 125.4 for the internal standard piracetam.SD rats were injected intravenously with single doses of 500,1 000 and 2 000 mg·kg-1 oxiracetam,respectively.The rat brain tissues were collected at different time intervals and the concentrations of oxiracetam in the brain tissues were determined.Results:The linear calibration curve of oxiracetam was obtained in the concentration range of 1.0-100.0μg·mL-1,and the lower limit of quantification(LLOQ)was 1.0 μg·mL-1. Both the intra-and inter-batch precisions of RSD were less than 5.7%.The accuracy was in the range of 96.3%-105.2%.The main pharmacokinetic parameters of oxiracetam in SD rats brain tissue after a single dose of 500,1 000,2 000 mg·kg-1 oxiracetam by intravenous route were as follows:t1/2=(3.56±0.27)h,(3.36±0.49)h,(3.60±0.79)h;Cmax=(24.78±16.06)μg·g-1,(50.79±25.92)μg·g-1,(82.21±19.33)μg·g-1;MRT0-t=(2.54±0.22)h,(2.38±0.30)h,(2.57±0.08)h;AUC0-t=(48.59±10.92)μg·h·g-1,(94.54±9.30)μg·h·g-1,(201.35±18.87)μg·h·g-1.Conclusion:The method can be used to investigate the pharmacokinetics of oxiracetam distributed in rat brain tissues.After being injected intravenously with a single dose of oxiracetam at 3 dosages,the uptake of oxiracetam into the brain tissue is very quick,maintained at a high concentration with a long duration.All the actions promote the activation and improvement of the brain functions.
作者
程强
羿国娟
叶雷
贺英
孙文霞
荣祖元
CHENG Qiang;YI Guo-juan;YE Lei;HE Ying;SUN Wen-xia;RONG Zu-yuan(Antibiotics Research and Re-evaluation Key Laboratory of Sichuan Province,Sichuan Industrial Institute of Antibiotic,Chengdu University,Chengdu 610052,China;Dongze Pharmaceutical Science and Technology Co.,Ltd.,Chengdu 400030,China;Sichuan Institute for Food and Drug Control,Chengdu 611731,China)
出处
《药物分析杂志》
CAS
CSCD
北大核心
2018年第11期1952-1959,共8页
Chinese Journal of Pharmaceutical Analysis