HPLC-荧光检测法测定犬血中紫花前胡苷的浓度及其药代动力学初步研究

目的:建立HPLC-荧光法测定犬血浆中紫花前胡苷的浓度,初步研究紫花前胡苷在犬体内的药代动力学特征。方法:以乙腈为蛋白沉淀剂,以Hypersil BDS C₁₈柱(200 mm×4.6 mm,5μm)为色谱柱,流动相为甲醇-0.1%磷酸溶液(45:55);激发波长为360 nm;发射波长为395 nm。Beagle犬4只,单次静脉注射紫花前胡苷30 mg·kg^-1,后肢静脉取血,采用HPLC测定血药浓度,并用DAS 2.0程序计算药代学参数。结果: Beagle犬血浆中内源性成分对测定无干扰,紫花前胡苷的线性范围为0.1~40.0μg·mL^-1(r=0.9993),平均相对回收率为89.8%~100.7%,日内和日间RSD均<10%。紫花前胡苷在犬体内的药代动力学过程符合开放二房室模型,主要药代动力学参数t_1/2α、t_1/2β、V₁、CL和AUC_(0-t) 分别为(0.536±0.085)h、(3.048±0.826)h、(0.446±0.038)L·kg^-1、(0.417±0.108)L·h^-1·kg^-1和(68.966±16.181) mg·L^-1·h。结论:本方法准确、灵敏、快速,适用于紫花前胡苷的血药浓度测定和药代动力学研究,为紫花前胡苷的开发和临床应用提供参考。

Objective: To establish a sensitive and reliable HPLC-fluorescence method to determine the concentration of nodakenin in Beagle dogs’ plasma,and to investigate its pharmacokinetics.Methods: The plasma samples were pretreated by acetonitrile for protein precipitation and were analyzed by HPLC.Separation and determination were achieved on a Hypersil BDS column(200 mm×4.6 mm,5 μm)with the mobile phase consisting of 0.1% phosphoric acid-methanol(55:45).The excitation wavelength and the emission wavelength were set at 360 nm and 395 nm,respectively.Four dogs were administered with nodakenin by intravenous injection at a single dose of 30 mg·kg^-1.The blood for test was taken from the vein on the posterior limb,and the plasma concentration was determined by HPLC.The main parameters of pharmacokinetics were calculated by DAS 2.0 software.Results: The endogenous ingredients in the Beagle dog blood showed no interference to the determination.The range of linearity was 0.1-40.0 μg·mL^-1(r=0.9993).And the mean recovery rate was 89.8%-100.7%.Both the intra-day and inter-day RSDs were less than 10%.The plasma concentration-time curve of nodakenin was conformed to be a two-compartment open model.The main pharmacokinetic parameters t_1/2α,t_1/2β,V₁,CL and AUC_(0-t) were(0.536±0.085)h,(3.048±0.826)h,(0.446±0.038)L·kg^-1,(0.417±0.108)L·h^-1·kg^-1 and(68.966±16.181)mg·L^-1·h,respectively.Conclusion: This method is accurate,sensitive,rapid,and suitable for plasma concentration determination and pharmacokinetic research of nodakenin,providing reference for its development and application.