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目的建立超高效液相色谱-串联质谱法(UHPLC-MS/MS)同时测定大鼠血浆中毛蕊花糖苷、异毛蕊花糖苷及连翘酯苷B的水平,并计算3个酚苷类成分在大鼠体内的药动学参数。方法大鼠ig给予裸花紫珠提取物(5 g/kg)后不同时间取血浆,血浆样品经盐酸处理,乙腈沉淀蛋白后,以乙腈-0.005%甲酸为流动相,通过Phenomenex®Kinetex C18柱(50 mm×2.1 mm,1.7μm)梯度洗脱;采用电喷雾离子化源(ESI)三重四极杆串联质谱,以多反应监测模式(MRM)负离子测定血浆中毛蕊花糖苷、异毛蕊花糖苷、连翘酯苷B。结果毛蕊花糖苷、异毛蕊花糖苷、连翘酯苷B分别在7.77~3 880.00 ng/m L(r~2=0.995 5)、5.04~2 520.00 ng/m L(r~2=0.994 9)、1.78~890.00 ng/m L(r~2=0.995 1)呈良好的线性关系,各成分的提取回收率在75.2%~89.9%,日内、日间精密度RSD均小于8.8%。大鼠ig给予裸花紫珠提取物后,毛蕊花糖苷、异毛蕊花糖苷及连翘酯苷B均在30 min左右达到最大血药浓度,AUC0~t分别为(93 881.65±18 326.65)、(29 204.97±8 499.88)、(15 027.05±3 763.82)ng·min/m L,Cmax分别为(2 179.00±355.60)、(737.57±210.31)、(227.30±48.38)ng/m L,t1/2z分别为(235.41±117.90)、(151.56±49.23)、(161.68±63.92)min。结论建立的UHPLC-MS/MS方法简便、快速、专属性强,可用于大鼠血浆中毛蕊花糖苷、异毛蕊花糖苷、连翘酯苷B的同时测定及其药动学研究。
OBJECTIVE To establish a method for the simultaneous determination of verbascoside, isogamidine and forsythiaside B in rat plasma by ultra performance liquid chromatography-tandem mass spectrometry (UHPLC-MS / MS), and calculate the levels of three phenolic glycosides in rats In vivo pharmacokinetic parameters. Methods The rat plasma was extracted with igneous flower extract (5 g / kg) at different time and the plasma samples were treated with hydrochloric acid. After the protein was precipitated by acetonitrile, acetonitrile-0.005% formic acid was used as mobile phase and passed through a Phenomenex® Kinetex C18 column (50 mm × 2.1 mm, 1.7 μm). Electrospray ionization (ESI) triple quadrupole tandem mass spectrometry (ESI-MS / MS) triple quadrupole tandem mass spectrometry was used to determine the concentrations of verbascoside and isoverine in plasma with multiple reaction monitoring mode (MRM) Caerulein B Results Verbascoside, Isoserbide and Forsythiaside B were in the range of 7.77 ~ 3880.00 ng / m L (r ~ 2 = 0.995 5), 5.04 ~ 20 520.00 ng / m L (r ~ 2 = 0.994 9) ~ 890.00 ng / m L (r ~ 2 = 0.995 1). The recoveries of all the components were in the range of 75.2% -89.9%. The RSDs of intra- and inter-day precision were less than 8.8%. After the rats were given igneous flower extract, Verbascoside, Isoserbide and Forsythiaside B reached the maximum blood concentration about 30 min, the AUC0 ~ t were (93 881.65 ± 18 326.65), (29 204.97 ± 8 499.88, 15 027.05 ± 3 763.82 ng · min / m L, respectively, and the Cmax values were (2 179.00 ± 355.60), (737.57 ± 210.31) and (227.30 ± 48.38) ng / (235.41 ± 117.90), (151.56 ± 49.23), (161.68 ± 63.92) min. Conclusion The established UHPLC-MS / MS method is simple, rapid and specific and can be used for the simultaneous determination and pharmacokinetics of verbascoside, isogamidine and forsythiaside B in rat plasma.