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,1.7 μm)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/mL(r²＝0.995 5)、5.04～2 520.00 ng/mL(r²＝0.994 9)、1.78～890.00 ng/mL(r²＝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,AUC_0~t分别为(93 881.65±18 326.65)、(29 204.97±8 499.88)、(15 027.05±3 763.82)ng·min/mL,C_max分别为(2 179.00±355.60)、(737.57±210.31)、(227.30±48.38)ng/mL,t_1/2z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 An UHPLC-MS/MS method was developed for the simultaneous determination of acteoside, isoacteoside, and forsythoside B in plasma of rats and the pharmacokinetic parameters for three phenolic glycosides were calculated as well. Methods Samples of plasmaof ratswere obtainedat different time after rats were administrated with Callicarpa nudifloraextract (5 g/kg).After the addition of acidification (hydrochloric acid, 0.25 mol/L) and deproteinization by acetonitrile, plasma samples were separated on a Phenomenex^® Kinetex C₁₈ column (50 mm ×2.1 mm, 1.7 μm) with gradient elution using acetonitrile-0.005% formic acid as mobile phase. Mass spectrometric detection was carried out by multiple reaction monitoring (MRM) using electrospray ionization in negative ion mode. Results A good linearity of acteoside, isoacteoside, and forsythoside B was shown in the ranges of 7.77 - 3 880.00 ng/mL (r² = 0.995 5), 5.04 - 2 520.00 ng/mL (r² = 0.994 9), and 1.78 – 890.00 ng/mL (r² = 0.995 1), respectively. The mean extraction recoveries of analytes were in the range of 75.2% - 89.9%, and the intra- and inter-day RSD values were less than 8.8%. The t_max ofacteoside, isoacteoside, and forsythoside Bwas about 30 min,AUC_0~t were (93 881.65 ±18 326.65), (29 204.97 ±8 499.88), and (15 027.05 ±3763.82) ng·min/mL, C_max were (2 179.00 ±355.60), (737.57 ±210.31), and (227.30 ±48.38) ng/mL, t_1/2z were (235.41 ±117.90), (151.56 ±49.23), and (161.68 ±63.92) min, respectively. Conclusion The method is proved to be simple, rapid, and specific, and to be suitable for the simultaneous determination of acteoside, isoacteoside, and forsythoside B in plasma of rats and the pharmacokinetic study.

国家自然科学基金资助项目(81373955);中国食品药品检定研究院中青年发展研究基金课题(2013WA9);江西省自然科学基金资助项目(20142BAB205085)