反相高效液相色谱法测定大鼠脑组织吗啡-3-葡萄糖醛酸苷含量  

DETERMINATION OF MORPHINE-3-GLUCURONIDE IN RAT BRAIN IN VITRO BY REVERSED-PHASE HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

作　　者：王丽[1] 阎本勇[1] 陈亦兰[1] 刘丽京[1] 孙莉[1] 邓艳萍[1] 

机构地区：[1]北京大学中国药物依赖性研究所,北京100191

出　　处：《中国药物依赖性杂志》2010年第1期25-28,共4页Chinese Journal of Drug Dependence

基　　金：国家科技重大专项课题(2009ZX09301-010);国家科技支撑计划项目(2008BAI49B01)资助

摘　　要：目的:建立一种反相高效液相色谱(RP-HPLC)检测法,用于测定大鼠脑组织中吗啡-3-葡萄糖醛酸苷(M3G)的浓度。方法:微粒体酶体外反应体系,经孵育和处理后,采用HPLC分析定量,并进行方法学考察。RP-HPLC测定采用DiamonsilTMC18色谱柱(250mm×4.6mm,5μm),流动相为10mmol.L-1磷酸二氢钾缓冲液(含1mmol.L-1十二烷基磺酸钠,磷酸调pH=2.15):乙腈(79.5∶20.5),流速:1.5ml.min-1,紫外检测波长:210nm,进样量50μl。结果:本实验条件下各物质分离良好,M3G线性范围50-600μg.L-1。M3G最低检测限为50μg.L-1。M3G的方法回收率在90%-105%范围内,提取回收率在75%以上。结论:本法可用于对大鼠脑微粒体体外孵育生成吗啡-3-葡萄糖醛酸苷含量的测定。Objective：To establish a reversed-phase high performance liquid chromatography（RP-HPLC）method for the determination of morphine＇s metabolite-M3G in rat brain.Methods：Chromatographic separation was achieved with a DiamonsilTM C18 analytical column（250 mm× 4.6 mm,5 μm）.The mobile phase was 10 mmol·L-1 potassium phosphate and 1 mmol·L-1 sodium dodecyl sulfate（pH=2.15 adjusted by phosphoric acid）-acetonitrile（79.5∶20.5）,and flow rate was 1.5 ml · min^-1.The drug concentrations were measured by UV at 210 nm.Results：The calibration curves were linear in the concentration range of 50-600 μg · L^-1 for M3G.The limit of quantitation（LOQ）was 50 μg· L-1 for M3G.The method recovery of the sample was within 90%-105%,the absolute recovery was above 75%.Conclusion：This method is simple and rapid,especially for quantitative analysis of M3G in rat brain in vitro.

关 键 词：反相高效液相色谱法 吗啡-3-葡萄糖醛酸苷(M3G) 吗啡代谢 微粒体 

分 类 号：R927.2[医药卫生—药学] R927.1