丹参酮Ⅱ_A在Beagle犬体内的血药浓度测定及其药动学研究

Study on the Plasma Concentration Determination and Pharmacokinetics of Tanshinone ⅡAin Beagle Dogs

目的：建立丹参酮ⅡA血药浓度测定方法,并用于丹参酮ⅡA在Beagle犬体内的药动学研究。方法：采用高效液相色谱（HPLC）法。色谱柱为Phenomenex Luna C18,流动相为水（5 mmol/L醋酸铵＋0.05%H3PO4）-乙腈,流速为1.0 ml/min,检测波长为270 nm,柱温为40℃,进样量为20μl。9只Beagle犬随机分为丹参酮ⅡA低、中、高剂量（2、4、8 mg/kg）组,分别于iv给药前及给药后2、5、10、15、20、30、45、60、75、90、120 min,从前肢静脉取血测定血药浓度,采用DAS 2.0软件计算药动学参数。结果：丹参酮ⅡA检测质量浓度线性范围为0.097 5~12.50μg/ml（r=0.999 8）,精密度、稳定性试验的RSD均小于10%,方法回收率为100.4%~107.2%,提取回收率为90.2%~92.3%。丹参酮ⅡA低、中、高剂量组t1/2α分别为（1.01±0.27）、（1.03±0.46）、（1.51±0.65）min,t1/2β分别为（16.25±4.78）、（22.42±9.32）、（24.45±12.02）min,AUC0-120 min分别为（150.88±45.25）、（305.44±92.55）、（643.67±178.27）μg·min/ml,CL分别为（12.01±4.36）、（12.78±5.06）、（12.17±5.41）ml/（min·kg）。结论：本方法精密度、稳定性和准确度均符合生物样品测定要求,丹参酮ⅡA在Beagle犬体内AUC0-120 min与剂量呈现良好的线性关系。

OBJECTIVE：To establish a method for determining plasma concentrations of tanshinone ⅡAand use it for pharmacokinetics study of tanshinone ⅡAin Beagle dogs.METHODS：HPLC was performed on the column of Phenomenex Luna C18,with the mobile phase of water（5 mmol/L of ammonium acetate＋0.05% H3PO4）-acetonitrile at flow rate of 1.0 ml/min;the detection wavelength was 270 nm,column temperature was 40 ℃ and the volume was 20 μl.9 Beagle dogs were randomly divided into tanshinone ⅡAlow,medium and high dose groups（2,4 and 8 mg/kg）,blood was respectively taken from forelimb venous blood to determine the plasma concentration before and after 2,5,10,15,20,30,45,60,75,90 and 120 min of iv administration.DAS 2.0 software was used to calculate the pharmacokinetic parameters.RESULTS：The linear range of tanshinone ⅡAwas 0.097 5-12.50 μg/ml（r=0.999 8）;the RSDs of precision and stability tests were all less than 10%;the method recovery was 100.4%-107.2%,and extraction recovery was 90.2%-92.3%.The t1/2αin tanshinone ⅡAlow,medium and high dose groups were respectively（1.01±0.27）,（1.03±0.46）and（1.51±0.65）min;t1/2βwere respectively（16.25±4.78）,（22.42±9.32）and（24.45±12.02）min;AUC0-120 minwere respectively（150.88±45.25）,（305.44±92.55）and（643.67±178.27）μg·min/ml;and CL were respectively（12.01±4.36）,（12.78±5.06）and（12.17±5.41）ml/（min·kg）.CONCLUSIONS：The precision,stability and recovery of the method are all in line with the determination requirements of biological samples,and tanshinone ⅡAshowed a good linear relationship with dose in Beagle dogs in AUC0-120 min.