摘要
目的:建立测定血清中靛玉红含量的高效液相色谱(HPLC)方法,并应用于靛玉红在大鼠体内的药代动力学研究。方法:采用ODS C18色谱柱(4.6mm×250mm,5μm),流动相为甲醇-水(75∶25),流速1.0mL·min-1,检测波长289nm,柱温35℃,进样量为20μL,内标物为炔雌醇。给大鼠颈静脉插管后,分别按2.0,4.0mg·kg-1经插管静脉注射靛玉红,用HPLC测定给药后不同时刻大鼠血清中靛玉红的浓度。采用Winnonlin5.0软件拟合,计算药动学参数。结果:靛玉红在0.031～2.48mg·L-1线性关系良好(R2=0.9996),最低检测限31μg·L-1。回收率大于98%,其日内、日间RSD均小于10%。靛玉红在大鼠体内的药代动力学过程符合二室模型。结论:本方法操作简便、准确,灵敏度高、重复性好,可用于靛玉红血药浓度的监测及其药代动力学研究。
Objective： To improve the method of indirubin in serum by HPLC and apply to pharmacokinetics in rats. Method： Chromatographic separation was conducted on an C18 column （4. 6 mm × 250 mm, 5 μm） , using a mixture of methanol-water （75：25） as mobile phase at a flow rate of 1.0 mL min^- 1 with UV detection at 289 nm, the column temperature was at 35 ℃ and ethinyl estradiol was used as an internal standard. Rats were administered i. v. bolus of indirubin in doses of 2.0 and 4. 0 mg · kg^-1 through a jugular vein catheter, respectively. Serial blood samples （ about 100 μL） were individually collected at 2, 5, 10, 20, 30, 60, 90, 120, 180 min after administration, and the concentrations of indirubin determined were in rat serum by HPLC. The pharmacokinetie parameters were calculated with the Winnonlin 5.0 software. Result： The calibration curve for indiruhin was linear （ R^2 = 0. 999 6） in the range of 0.031-2. 48 mg · L^-1 and the limit of detection （LOD） was 31 μg · L^-1. The average recovery of indirubin in rat serum was more than 98% and the relative standard deviations of intra-day and inter-day were both less than 10%. The pharmacokinetics of Indirubin in rats was fitted to two-compartment model. Conclusion： The method is simple and accurate with a high sensitivity and a good repeatability, and it can be applied to the evaluation of pharmacokinetic parameters of indirubin in rats and blood concentration of indirubin in clinical controlling.
出处
《中国中药杂志》
CAS
CSCD
北大核心
2010年第9期1148-1151,共4页
China Journal of Chinese Materia Medica
基金
北京市属市管高等学校人才强教计划资助项目
