HPLC-MS/MS测定血浆中丁丙诺啡的浓度

Determination of Buprenorphine in Human Plasma by Liquid Chromatography-Tandem Mass Spectrometry

目的建立人血浆中丁丙诺啡浓度的HPLC-MS/ MS 测定法。方法以曲马多为内标,将待测血浆样品经氢氧化钠碱化后,加正己烷提取,上清液经Agilent Eclipse plus C18 （4. 6 mm ×150 mm,3. 5 μm）色谱柱分离,甲醇- 水（80∶20,v∶v,含0. 005 mol/ L 甲酸铵和0.25%甲酸）为流动相,流速0. 5 mL/ min,进行洗脱,采用电喷雾电离源（ESI）,以多反应监测（MRM）方式进行正离子检测。用于定量分析的离子分别为m/ z 468. 3→55. 2 和m/ z 264. 2→58. 1（内标）。结果血浆中丁丙诺啡的浓度与峰面积比在50-20 000 ng/ L 的范围内线性关系良好,定量下限为50 ng/ L;提取回收率大于67%,日内日间精密度小于11%。结论该法简单、灵敏、快速,分离效果良好,可用于人体血药浓度测定和药动学研究。

Objective To develop an HPLC-MS/MS method for the determination of buprenorphine in human plasma. Methods Plasma were alkalified with sodium hydroxide, extracted with n-hexane, and then separated on a Eclipse plus C18 （4. 6 mm ×150 mm,3. 5 μm） column with tramadol as internal standard. The mobile phase was composed of methanol and ammonium formate-formic acid buffer containing 0. 005 moL/L ammonium formate and 0.25 % formic acid （80： 20,v/v）, at a flow rate of 0. 5 mL/min. Multiple reaction monitoring （MRM） transition of m/z 468.3→55.2 and m/z 264.2→58. 1 was performed in positive mode. Results The calibration curves of buprenorphine were linear in the range of 50-20 000 ng/L. The lower limit of quantification was 50 ng/L. The recovery was more than 67%. The intra and inter-day precision were less than 11%. Conclusion The method is simple, quick and sensitive, and it is suitable for concentration determination of buprenorphine in plasma and pharmacokinetics study.