LC-MS／MS法测定人血浆中小檗碱的浓度及其药动学研究(英文)  

LC-MS/MS method for quantitation of berberine in human plasma:application to a pilot pharmacokinetic study in healthy Chinese subjects

作　　者：张丹[1] 韩静[1] 王晓琳[1] 王涛[1] 魏振满[2] 刘会臣[1] 

机构地区：[1]航天中心医院临床药理室,北京100049 [2]中国人民解放军302医院临床药理室,北京100039

出　　处：《中国新药与临床杂志》2013年第2期123-129,共7页Chinese Journal of New Drugs and Clinical Remedies

摘　　要：目的建立测定人血浆中小檗碱的液相色谱串联质谱(LC-MS/MS)法。方法六名健康受试者单次口服盐酸小檗碱片(含盐酸小檗碱100 mg)后,采集血样,并用LC-MS/MS法测定血浆中小檗碱的浓度。血浆样本经液-液萃取法处理,选用Inertsil ODS-SP色谱柱(75 mm×2.1 mm,3μm),以乙腈:10 mmol·L-1乙酸铵(含0.1%甲酸)(55:45,V/V)为流动相,流速为0.3 mL·min-1。选用API 3200型三重四极杆串联质谱仪的多重反应监测(MRM)扫描方式进行监测,电喷雾离子化源,正离子方式。结果小檗碱以及内标克拉霉素的保留时间分别为1.17、1.14 min;血浆中小檗碱的线性范围为0.005 00～2.00μg·L-1(r>0.99),定量下限为0.005 00μg·L-1;日内、日间相对标准差(RSD)均小于12.0%;相对误差(RE)均在±14.0%的范围以内;提取回收率较高,且可重现;小檗碱在各种贮存条件下均较稳定。该方法成功地应用于盐酸小檗碱片在中国健康人体内的药动学研究。六名健康受试者单次口服盐酸小檗碱片(含盐酸小檗碱100 mg)后,ρmax、AUC0-t和t1/2值分别为0.061 8～0.354μg·L-1、0.906～6.04μg.h.L-1和6.4～56.3 h。结果表明口服盐酸小檗碱片后小檗碱的吸收程度较低,并且其主要药动学参数存在较大的个体间差异。结论该方法快速、灵敏、专属性强、重现性好,适用于小檗碱的人体药动学研究。AIM To develop a rapid and sensitive liquid chromatography- tandem mass spectrometric （LC-MS/MS） method for the quantitation of berberine in human plasma. METHODS Six healthy subjects were given a single oral dose of berberine hydrochloride tablet containing 100 mg berberine hydrochloride. Blood samples were collected at designated time points. Plasma concentrations of berberine were quantified by a selective and sensitive liquid chromatography-tandem mass spectrometric （LC-MS/MS） method. After extracted from plasma by liquid-liquid extraction, berberine and clarithromycin （internal standard, IS） were separated on an Inertsil ODS-SP column （75 mm × 2.1 mm, 3μm） using acetonitrile-10 mmol＇L-t ammonium acetate （containing 0.1% formic acid） （55 ： 45, V/V） as mobile phase at a flow rate of 0.3 mL.min-1. Detection was by electrospray positive ionization mass spectrometry using multiple reaction monitoring （MRM） mode. RESULTS Berberine and IS were eluted at 1.17 and 1.14 min, respectively. The method was linear over the concentration range of 0.005 00 - 2.00 Ixg. L-I for berberine with a lower limit of quantitation of 0.005 00 Ixg＂ L-l. The intra- and inter-day precision as relative standard deviation （RSD） were 〈 12.0%, and accuracy as relative error （RE） were within ＋ 14.0%. The extraction recoveries were high and reproducible. Berberine was found to be stable under various storage conditions. The method was successfully applied to a pilot pharmacokinetic study of berberine in healthy Chinese subjects after a single oral dose of berberine hydrochloride tablet containing 100 mg berberine hydrochloride, in which the values of ρmax, AUC0 t and tl/2 were 0.061 8 - 0.354 μg.L-1, 0.906 - 6.04μg.h.L-1 and 6.4 - 56.3 h, respectively. The results indicated the absorption rate of berberine was poor, and there was significant individual difference in the pharmacokinetics of berberine in healthy Chinese subjects after a single oral dose. CONCLUSION The established method is rapi

关 键 词：小檗碱 色谱法 液相 串联质谱法 血浆 药动学 

分 类 号：R917[医药卫生—药物分析学] R969.1[医药卫生—药学]