HPLC-MS/MS测定人体内齐墩果酸血药浓度

 目的 建立快速、灵敏的高效液相色谱-串联质谱（HPLC-MS/MS）法测定人血浆中的齐墩果酸。方法 血浆样品定量加入内标液，经乙酸乙酯液液萃取后，以甲醇-0.1%醋酸铵缓冲溶液（93∶7）为流动相，采用Zorbax Extend-C₁₈（4.6 mm×250 mm， 5 μm）柱分离。使用电喷雾离子化，负离子SRM检测。用于定量的离子反应分别为m/z 455.1→455.3（齐墩果酸）；m/z 469.1→469.3（内标甘草次酸）。结果 测定血浆中齐墩果酸的线性范围为0.1～40.0 μg·L-1，定量限为0.1 μg·L-1，批内、批间精密度（RSD）均小于20.0%，方法学回收率94.0%～102.0%，符合相关规定。单个样品的分析时间为6.5 min。本法已用于齐墩果酸口服制剂的人体生物等效性研究。结论 本方法灵敏度高，选择性强，分析测试时间较短，适用于人血浆样品中齐墩果酸的测定。

HPLC-MS/MS Determination of Oleanolic Acid in Human Plasma

OBJECTIVE To develop and validate a method for the determination of oleanolic acid in human plasma by HPLC-MS/MS. METHODS The plasma sample was extracted with ethyl acetate， then separated on a Zorbax Extend-C₁₈（4.6 mm×250 mm， 5 μm）column using methanol-0.1%ammonium acetate (93∶7) as the mobile phase. The triple quadrupole mass spectrometry was applied. Quantitation was performed using positive ion selected reaction monitoring (SRM) of the transitions of m/z 455.1→455.3 and m/z 469.1→469.3 for oleanolic acid and the internal standard glycyrrhetinic acid， respectively. RESULTS The linear calibration curves were obtained in the concentration range of 0.1-40.0 μg·L-1. The limit of quantification was 0.1 μg·L-1. The intra-day and inter-day relative standard deviation over the entire concentration range was below 20.0%. The accuracy was 94.0%-102.0%. Each sample was finished within 6.5 min. The method was applied to evaluate the bioequivalence of two oleanolic acid tablet formulations in healthy volunteers. CONCLUSION This method is sensitive， selective， rapid and suitable for the determination of oleanolic acid in human plasma.