【摘要】 目的:通过细胞摄取巴氯酚的体外药动学研究及液质联用定量检测方法的建立,为确定巴氯酚更合理的给药途径和跨越血脑屏障的机制研究提供依据。方法:利用CHO-K1细胞摄取药物的细胞生物学实验方法结合液质联用检测技术,以细胞/介质(cell/medium,C/M)比值为指标进行CHO-K1细胞摄取巴氯酚体外药动学研究,并进行方法学考察。本实验选取的摄取时间点分别为30s,1,5,30,60min。结果:液质联用定量方法灵敏度高,重复性好。在0～30min区间内,细胞摄取巴氯酚的量随时间几乎呈线性增加,30min时的C/M值达到最高为9.74(n=4),基本达到饱和状态,30～60min区间,细胞摄取巴氯酚的量几乎不再随时间发生变化。结论:为深入研究巴氯酚跨越血脑屏障的机制并确定实际应用的给药途径提供数据基础和参考依据。定量方法简便快捷,灵敏度高,重复性好,可广泛应用于细胞摄取微量药物的检测研究中,具实用价值。更多还原

【Abstract】 OBJECTIVE To study the mechanism of baclofen crossing the blood-brain barrier and to find out a more effective route of administration for baclofen,the time-course experiment for baclofen uptake by CHO-K1 cells was done and the content of baclofen in samples was determined by LC-MS/MS technology.METHODS By the means of the cell uptake experiment method together with LC-MS/MS technology and using cell/medium ratio as evaluation criteria to study the transport properties of uptake of baclofen by CHO-K1 cells.Meanwhile,the methodology was investigated.Time points were chosen as 30 s,1 min,5 min,30 min and 60 min,respectively.RESULTS The method of determination by LC-MS/MS technology was high sensitive and better reproducibility.The results showed that the uptake of baclofen by CHO-K1 cell increased nearly lineally with time from 0min to 30min and the C/M ratio was 9.74(n=4)at 30 min,the uptake was nearly saturated and there was no significant change from 30min to 60 min.CONCLUSION This experiment offered the data to study further about the mechanism of baclofen crossing the blood-brain barrier and investigate much more effective route of administration of baclofen.The method of determination by LC-MS/MS technology was high sensitive and better reproducibility and practicable and was able to be applied in the study of compounds uptake by cells.更多还原