高效液相色谱测定急性胰腺炎香猪用药后血中大黄素及汉防己甲素

Measurement of Emodin and Tetrandrine in the Blood of Miniature Pigs with Acute Pancreatitis by HPLC after Using Chinese Medicines

目的 用高效液相色谱检测急性胰腺炎 (acutepancreatitis ,AP)香猪在应用清胰汤 (Qing yi tang ,QYT)及汉防己甲素 (Tetrandrine ,Tet)后血清大黄素与血浆Tet的含量 ,为药效学及药代动力学的进一步研究提供方法学的基础。方法 采用胰管逆行加压推注牛磺脱氧胆酸钠复制AP香猪模型 (n =18) ,将动物随机均分 3组 :QYT组 (AP+QYT)、Tet组 (AP +Tet)、联合用药组 (AP +QYT +Tet) ,分别接受相应药物 ,每 12h 1次 ,连续 3d。于制模前、制模后 2 4、48、72h同时采门静脉血和腔静脉血 (每次采血均在前一次用药后 12h) ,分离血清或血浆 ,用高效液相色谱检测血清大黄素及血浆Tet的含量。结果 大黄素在 5 0～ 2 5 0ng/ml含量范围内、Tet在 5 0～ 40 0ng/ml含量范围内与相应的峰值呈良好的线性关系 ,相关系数分别为 0 .994,0 .993;回收率分别为 84.6 %和 89.1%。AP +QYT组的 2 4、48、72h的门静脉血清大黄素含量较稳定 ;腔静脉的在 2 4、48h稍低 (P >0 .0 5 ) ,72h的含量较高 ,差异显著 (P <0 .0 5 )。AP +Tet组的 2 4、48h腔静脉血浆Tet含量高于门静脉的 ,以 48h的为明显 (P <0 .0 5 ) ;72h的测定值也高于其他时间点 (P <0 .0 5 )。AP +QYT +Tet联合用药组在各时间点 。

Objective By high performance liquid chromatography(HPLC),serum emodin and plasma tetrandrine levels were tested to reflect the blood concentrations of Chinese medicine,Qing-Yi-Tang(清胰汤,QYT) and tetrandrine(Tet) used in treating acute pancreatitis(AP).Methods 18 miniature pigs were fasted over night,and then subjected to laparotomy.The portal vein and vena cava were catheterized.Model pigs of AP were induced by a retrograde injection of 5% sodium taurocholate into the bilio-pancreatic duct,and then divided into three groups:one was treated with QYT(AP+QYT),one with Tet(AP+Tet),and one with QYT+Tet(AP+QYT+Tet).Tet was injected intraperitoneally,and QYT was given with a nose-gastric catheter.These medicines were given within 10 min,and repeated every 12 h after AP inducement.At the pre-model time and 24 h,48 h,72 h after AP,the blood samples were taken from both portal vein and vena cava.Emodin in the serum,and Tet in the plasma were tested by HPLC.Results By HLPC,the linear concentration range of emodin was from 50 ng·ml -1 to 250 ng·ml -1,and Tet was from 50 ng·ml -1 to 400 ng·ml -1.Their average recovery rates were 84.6% and 89.1% respectively.The serum emodin in AP+QYT group was stable at 24 h and 48 h,and increased at 72 h after AP.The plasma Tet levels in AP+Tet group showed a similar change style as that in AP+QYT.In AP+QYT+Tet group,the serum emodin levels were not obviously changed,but the plasma Tet levels increased significantly as compared with AP+Tet group.Conclusion HLPC used in biomedical sample analysis of emodin and Tet is simple,rapid and accurate. The increased Tet levels in the AP+QYT+Tet group may be one of the mechanisms of the synergetic effect of these medicines.