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目的 建立并验证一种快速测定大鼠血浆中鸡骨草皂苷β-D-吡喃葡萄糖-3-(O-β-D-吡喃葡萄糖氧基)-齐墩果酸(AA-2)浓度的超高效液相串联三重四级杆质谱检测法(UPLC-MS/MS).方法 取大鼠适量全血,置肝素化离心管中,离心分离出血浆.氯唑沙宗(IS)作为内标,用乙腈-甲醇溶液(9∶1)进行蛋白质沉淀后,用UPLC BEH C18 Column (2.1mm×100 mm,1.7 μm)色谱柱,以含0.1％的甲酸水和乙腈进行梯度洗脱.用电喷雾离子源负离子模式下的多反应监测模式(MRM)进行测定,AA-2和氯唑沙宗的离子对分别为m/z 793.5→631.4、m/z168.1→132.1.结果 AA-2在10～5000 ng·mL-1内线性良好,其回收率为70.4％ ～ 87.8％,日间和日内精密度均＜10％,准确度为91.1％～104.1％.大鼠静脉给予3个浓度(1,3,5 mg·kg-1)AA-2后,MRT0-t分别为(0.32 ±0.13),(0.48±0.15),(0.44±0.08) h;t1/2分别为(1.15±0.72),(1.32±0.90),(1.02±0.22)h;CL分别为(4.53±1.66),(4.44±0.94),(4.86±0.49)L·h-1·kg-1,Cmax分别为(681.67±246.78),(1809.45 ± 479.11),(4953.21 ± 722.72)ng·mL-1.结论 该方法操作简便、快捷,灵敏度高,适于大鼠体内AA-2的药代动力学研究.“,”Objective To determine the concentration ofβ-D-gluco-pyranosy1-3-(0-β-D-glucopyranosyloxy)-oleanolate (AA-2) in rat plasma by ultra performance liquid chromatography and tandem mass spectrometry (UPLC-MS/MS) method.Methods Plasmas were separated from the heparinized whole blood by centrifugation.After addition of chlorzoxazone as an internal standard (IS),protein precipitation by acetonitrile-methanol (9∶1) was used to prepare samples.Chromatographic separation was achieved on a UPLC BEH C18column (2.1 mm × 100 mm,1.7 μm) with 0.1％ formic acid and acetonitrile as the mobile phase with gradient elution.An electrospray ionization source was applied and operated in negative ion mode.Multiple reactions monitoring (MRM) mode was used for quantification using target fragment ions m/z 793.5→631.4 for AA-2,and m/z 168.1→132.1 for IS.Results Calibration plots were linear throughout the range of 10-5000 ng · mL-1 for AA-2 in rat plasma.Mean recoveries of AA-2 in rat plasma ranged from 70.4％ to 87.8％.RSD of intra-day and inter-day precision were both less than 10％.The accuracy of the method was between 91.1％ and 104.1％.After intravenous administration of AA-2(1,3,5 mg · kg-1),the mean residence time(MRT0-t) were (0.32 ±0.13),(0.48 ±0.15),(0.44±0.08) h.The i1/2 were (1.15±0.72),(1.32 ± 0.90),(1.02 ± 0.22) h.The CL were (4.53±1.66),(4.44 ±0.94),(4.86±0.49) L · h-1 · kg-1 The Cmax were (681.67 ±246.78),(1809.45 ±479.11),(4953.21 ± 722.72) ng· mL-1.Conclusion The method was simple,rapid and sensitive,which was suitable for pharmacokinetics study of AA-2 after intravenous administration in rats.