UHPLC-MS/MS同时测定大鼠血浆中裸花紫珠3个酚苷类成分及其药动学研究  

Simultaneous determination of three phenolic glycosides in Callicarpa nudiflora by UHPLC-MS methods and analysis of their pharmacokinetics in plasma of rats

作　　者：郑东昆 陈伟康 马双成[3] 邵军[1,2] 王杰[1,2] 罗跃华[1,2] 

机构地区：[1]江西省药品检验检测研究院/江西省药品与医疗器械质量工程技术研究中心,江西南昌330029 [2]南昌大学药学院,江西南昌330006 [3]中国食品药品检定研究院,北京100050

出　　处：《中草药》2015年第23期3533-3538,共6页Chinese Traditional and Herbal Drugs

基　　金：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)

摘　　要：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 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 plasma of rats were obtained at different time after rats were administrated with Callicarpa nudiflora extract（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#174; 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/m L（r-2 = 0.995 5）, 5.04-2 520.00 ng/m L（r-2 = 0.994 9）, and 1.78 – 890.00 ng/m L（r-2 = 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 tmax of acteoside, isoacteoside, and forsythoside B was about 30 min, AUC0-t were（93 881.65 ± 18 326.65）,（29 204.97 ± 8 499.88）, and（15 027.05 ± 3763.82） ng·min/m L, Cmax were（2 179.00 ± 355.60）,（737.57 ± 210.31）, and（227.30 ± 48.38） ng/m L, t1/2z 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.

关 键 词：裸花紫珠 毛蕊花糖苷 异毛蕊花糖苷 连翘酯苷B 超高效液相色谱-串联质谱法 药动学 

分 类 号：R285.51[医药卫生—中药学]