关木通药材中马兜铃酸A的药动学研究

Study on pharmacokinetics of aristolochic acid caulis aristol ochiae Manshuriensis extraction

目的采用高效液相色谱法测定血浆中马兜铃酸A的浓度,研究小鼠灌胃给予关木通提取液后体内马兜铃酸A的药动学特点.方法采用DiamonsilTMC18色谱柱(250mm ×4.6mm,5μm),以甲醇-水-冰乙酸(72:27:1)为流动相,流速1.0mL·min-1,检测波长315nm.小鼠按马兜铃酸A 4.0mg·kg-1灌胃给予关木通提取液后,经HPLC法测定血浆药物浓度.结果马兜铃酸A在0.1098～3.66mg·L-1的浓度范围内线性关系良好(r=0.9998,n=7),最低检测浓度为0.0183mg·L-1.马兜铃酸A高、中、低3种不同浓度的平均回收率及相对标准偏差分别为97.07%(1.0%),97.10%(1.1%)和99.36%(0.8%),该法的日内和日间精密度RSD值均小于9.3%.小鼠给药后,药动学房室模型为一室模型,主要药动学参数T1/2(ka),T1/2(ke),Tmax,AUC,Cmax,CL/F(s),V/F(c)分别为:3.94,16.29,10.65min,188.88(mg/L)·min,2.84mg/L,0.038mg/kg/min/(mg/L),0.89(mg/kg)/(mg/L).结论本实验建立了关木通药材中马兜铃酸A血药浓度的HPLC测定方法,阐明了马兜铃酸A的药动学特征.

OBJECTIVE To establish a revered-phase HP LC method for the study of pharmacokinetics of aristolochic acid A from Caulis Aristolochiae manshuriensis extration in mice after single oral administratio n.METHOD The determination of aristolochic acid A concentration w as performed by HPLC on a waters apparatus with a 315nm ulerstraviolot detection and Diamonsil TM C 18 column (250mm×4.6mm, 5μm), which was eluted w ith a mixture of methanol-water-acetic acid (72∶27∶1) solution at a flow rat e of 1.0mL·min -1 and at a room temperature of 24℃. Mice were given the e xtraction by oral administration at dose of 4.0mg/kg aristolochic acid A and pla sma concentrations were determined by HPLC.RESULTS The standard curve showed a good linearity over a range of 0.1098~36.6mg·L -1 for aristolochic acid A (r=0.9998, n=7). The lo west determination concentration of aristolochic acid A was 0.0183mg·L -1. The average recoveries and relative standard deviations of aristolochic acid A were 99.36% (0.8%), 97.10% (1.0%), and 97.08% (1.0%), respectively. The relative standard deviations of intra-day and inter-day precisions for the method were all less than 9.3%. After single oral administration of the extraction to mice, the mean plasma concentration-time course was found to fit a one-compartment model. The main pharmacokinetic parameters of aristolochic acid A as follows: T 1/2(ka), T 1/2(ke), T max, AUC, C max, CL/F (S), V/F (c) were 3.94min, 16.29min, 10.65min, 104.88(mg/ L)·min, 2.84mg/L, 0.038mg/kg/min/(mg/L), 0.89(mg/kg)/(mg/L). CONCLUSION It is a good method to determine the concentration o f aristolochic acid A in plasma and to obtain pharmacokinetic parameters.