摘要
建立了山羊血浆中三苯双脒的高效液相色谱检测方法。山羊血浆用氢氧化钠和三乙铵处理,再用甲苯萃取三苯双脒,高速离心后,上清液真空干燥,残留物用含内标物的甲醇溶解后分析。采用WatersC18色谱分析柱(3.9mm×150mm,10μm),流动相为甲醇 水(50∶50,pH3),流速0.6mL/min,UV检测波长249nm,柱温为25℃,内标物为水杨酸(5μg/mL)。测得三苯双脒保留时间为4.3min,水杨酸保留时间为8.0min。在50～1000ng/mL浓度范围内三苯双脒和内标物峰面积之比与三苯双脒浓度具有良好的线性关系,回归方程为A=1456.67C 60.01(r=0.9747,n=5),RSD为5.1%。1000、500和100ng/mL质控浓度添加样品的日内(n=5)和日间(n=5)RSD分别为3.8%、9.7%、11.7%和5.2%、8.0%、11.1%。回收率为(94.3±1.1)%、(91.9±0.9)%和(94.7±5.0)%。最低检测限为20ng/mL。本方法操作简便,灵敏度高,为三苯双脒兽用制剂临床前及临床药物代谢动力学研究提供了分析方法。
A HPLC method for the determination of tribendimidin in goat plasma was established. The goat plasma(1 mL) was mixed with NaOH and triethylamine, then tribendimidin was extracted with toluene. After centrifugation, the supernatant liquids were evaporated to dry. The residues were dissolved in methanol with internal standard of salicylic acid and analyzed. Tribendimidin was separated on Waters C_(18) analytic column(3.9 mm×150 mm, 10 μm) at 25 ℃ with a methanol-water (50∶50) as mobile phase and detected with UV detector at 249 nm. The flow rate was 0.6 mL/min. The retention time of tribendimidin and salicylic acid was 4.3 min and 8.0 min, respectively. Calibrated curve in goat plasma was linear(r=0.974 7,n=5) from 50 to 1 000 ng/mL for tribendimidin. For the sample of tribendimidin 1 000, 500,100 ng/mL, the RSD of intra-run (n=5) and inter-run (n=5) were 3.8%, 9.7%, 11.7% and 5.2%, 8.0%, 11.1%, respectively, the average recoveries were ((94.3)±1.1)%,(91.9±0.9)%,(94.7±5.0)%, respectively. The limit of detection was 20 ng/mL. The analytical method for tribendimidin was sensitive, rapid and simple, suitable for application in preclinical and clinical pharmacokinetics studies.
出处
《中国兽药杂志》
2004年第9期17-19,共3页
Chinese Journal of Veterinary Drug
