基于UPLC-MS分析大鼠口服线叶菊有效部位后血浆中3种有效成分的药代动力学

Pharmacokinetics of Three Active Components in Rat Plasma After Oral Administraion of Effective Part of Filifolium sibiricum by UPLC-MS

目的:建立高效液相色谱-质谱法(UPLC-MS)测定大鼠口服线叶菊有效部位后血浆中异荭草素、异鼠李素-3-O-β-D-葡萄糖苷和东莨菪苷的含量,研究这3个指标性成分在大鼠体内的药代动力学行为。方法:采用ACQUITY UPLC HSS T3色谱柱(2.1 mm×100 mm,1.7μm),流动相0.1%甲酸水溶液(A)-0.1%甲酸乙腈溶液(B)梯度洗脱(0~2 min,98%A;2~6 min,98%~30%A;6.0~7.6 min,30%~40%A;7.6~8.0 min,40%~2%A;8~10 min,2%A;10~12 min,2%~98%A),流速0.4 m L·min-1,柱温40℃;电喷雾离子源(ESI),多反应监测(MRM)模式。采用单次给药法,大鼠按1.0 g·kg-1灌胃给予线叶菊有效部位,通过负离子模式测定血浆中异荭草素、异鼠李素-3-O-β-D-葡萄糖苷和东莨菪苷的含量。结果:各成分的线性关系良好(R2>0.997),各成分的提取回收率在80.87%~93.75%;日内、日间精密度RSD均<15%。3种成分的达峰时间(tmax)分别为(32.40±4.16),(30.00±4.80),(20.10±5.10)min,药峰浓度(Cmax)分别为(31.80±5.46),(28.00±2.54),(20.33±1.17)μg·L^(-1),药时曲线下面积(AUC0-t)分别为(37.80±3.94),(48.83±3.05),(32.40±2.64)μg·min·L^(-1),消除半衰期(t1/2β)分别为(114.06±22.19),(126.12±19.38),(148.14±24.10)min。结论:建立的UPLC-MS精密、准确,能够同时测定生物样品中异荭草素等3种成分的血药浓度。这3种成分进入体内及吸收的速度较快,但吸收情况并不理想。

Objective： An UPLC-MS method was developed for the simultaneous determination of isoorientin, isorhamnetin-3-O-13-D-glucoside and scopolin in plasma of rats after oral administration of effective part of Filifolium siburicum and the pharmacokinetie parameters of three components were calculated as well. Method ： Concentrations of the three analytes in plasma were separated on a ACQUITY UPLC HSS T3 column （2. 1 mm ×100 mm, 1.7 μm） with mobile phase of 0.1% formic acid aqueous solution-acetonitrile （containing 0. 1% formic acid） for gradient elution. Mass spectrometric detection was carried out by multiple reaction monitoring （MRM） with electrospray ionization source under negative ion mode. Single dose method was adopted, rats were oral treated with 1.0 g·kg^-1 of effective part of F. siburicum. Result： All calibration curves showed good linearity （Ra 〉 0. 997） over a wide concentration range for all analytes. The mean extraction recoveries of analytes were in the range of 80.87%-93.75% , and the intra-day and inter-day RSD were 〈 15%. The peak times （tmax） of isoorientin, isorhamnetin-3-O-β-D-glucoside and scopolin were （32.40 ± 4. 16）, （30.00 ± 4. 80）, （20. 10 ± 5. 10） min; the areas under the curve （AUC0-t） of them were （37.80 ± 3.94）, （48.83 ± 3.05）, （32.40 ± 2.64） μg·min·L^-1; the peak concentrations （Cmax） of them were （31.80 ±5.46）, （28.00 ±2.54）, （20.33 ± 1.17） txg.L-1 ; the elimination half-life （t1/2β） of them were （114.06 ±22. 19） , （126. 12 ± 19.38） , （148. 14 ±24. 10） min, respectively. Conclusion： This UPLC-MS is proved to be precise and accurate, and it is suitable for the simultaneous determination of blood concentration of isoorientin and other two components in biological samples. Rates of these three components into the body and the absorption are fast, but the absorption is not ideal.