盐酸安他唑啉在大鼠体内的药动学研究

Determination of antazoline hydrochloride in rat plasma by HPLC and its application to pharmacokinetic study

目的：建立高效液相色谱分析法评价灌胃给予盐酸安他唑啉在大鼠体内的药动学特征。方法：采用HPLC-内标法，液液萃取SD大鼠血浆中的待测物和内标物，以C18柱将待测物分离，用可变波长紫外检测器进行检测。流动相为甲醇-5mmol·L-1四丁基溴化铵（40：60）；流速为1．0ml·min。结果：盐酸安他唑啉在50-4000ng·ml-1范围内线性关系良好（R〉0．9993），定量下限达到50ng·ml-1。日内和日间精密度值均小于15％。准确度在89．9％-110．6％的范围内。盐酸安他唑啉的样品绝对回收率大于80％，非那西汀的绝对回收率在75％以上。灌胃后其主要动力学参数：AUG0-1=（5．15±0．39）mg·h·L-1、AUC=（5．37±0．26）mg·h·L-1、z1／2=（3．91±0．88）h、CL=（1．86±0．09）L·kg-1·h-1、V=（8．07±2．51）L·kg-1、c=（3．35±0．21）mg·L-1、tmax=（0．75±0．36）h。结论：该方法准确度高、灵敏度强，可用于大鼠灌胃给药盐酸安他唑啉后体内药物动力学研究。结果盐酸安他唑啉在大鼠体内消除快，不易蓄积，为其临床药动学以及生物等效性研究提供了理论依据。

OBJECTIVE To establish a HPLC method for the determination of antazoline hydrochloride in rat plasma and to study the pharmacokinetics of antazoline hydrochloride in rats. METHODS A sensitive and specific HPLC method was developed and validated using phenacetin as the internal standard （IS）. The analyte and the IS were extracted from rat by ethyl acetate under the basic condition. The analytes were separated by a C18 column and detected with a variable wavelength UV-detector. The mobile phase consisted of methanol-5 mmol L-1 tetrabutyl ammonium bromide （40：60） and the flow rate was 1.0 ml · min- 7 RESULTS Standard calibration graph for antazoline hydrochloride was linear over a curve range of 50 - 4 000 ng. ml- 1 （R= 0. 999 3） and the lower limit of quantification was 50 ng.m1-1 using a plasma sample of 100μ1. The intra- and inter-day precision values were less than 13.30% relative standard deviation （RSD）. The intra-day and inter-day assay precision was in the range of ＋ 15%and the accuracy in the range of 89. 9%- 110. 6%. The extraction recoveries were on the average of 80% for antazoline hydroehloride and 75% for IS. After i. g. administration of antazoline hydroehloride, the major pharmacokinetic parameters were as follows：AUGht = （5.15 ＋ 0. 39） mg.h.L-1 , AUC = （5.37 ±0. 26） mg.h-L-1 , t1/2 = （3.91 ± 0. 88） h, CL=（1.86±0.09） L.kg-1,h-1,V=（8.07±2.51） L-kg -1, C = （3. 35 ± 0. 21） mg-L-1 , tmax = （0. 75 ± 0. 36） h. CX）NCLUSION This method was successfully applied to pharmacokinetic studies of antazoline hydrochloride after ig administration to SD rats.