鸡血浆中乙酰氨基阿维菌素HPLC检测法的建立及口服给药后的药代动力学研究

Establishment of a HPLC Method for determination of eprinomectin in chicken plasma and the pharmacokinetics of eprinomectin in chicken following oral administration

关键词:  荧光高效液相色谱  乙酰氨基阿维菌素  鸡  药代动力学

Key words:  High performance liquid chromatography with fluorescence detecion  Eprinometin  Chicken  Pharmacokinetics

摘要:

旨在建立鸡血浆中乙酰氨基阿维菌素（EPR）浓度的高效液相色谱（HPLC）检测方法，并进行EPR在鸡体内的药代动力学研究。用甲醇提取血浆中的EPR，并用Sep-Pak C18固相萃取法进行纯化，纯化后的EPR经干燥处理，用三氟乙酸酐和N-甲基咪唑对其进行衍生化，使用荧光HPLC检测。结果表明，在血浆EPR含量为0.1~100 ng/mL范围内，标准曲线线性关系良好，相关系数r=0.9999。检测限（LOD）为0.1 ng/mL，定量限（LOQ）为0.3 ng/mL。批内批间的平均回收率均大于90%，批内变异系数2.81~8.02%，批间变异系数4.32~5.83%。给蛋鸡口服5.0 mg/kg的EPR，EPR在鸡体内的药代动力学参数显示：给药后1.58 h可达最大血药浓度（Cmax）354.27 ng/mL；消除半衰期（T1/2el）为5.52 h；平均滞留时间（MRT）为6.40 h。以上结果说明建立的检测方法灵敏度高、准确度高，干扰少，适用于鸡血浆中EPR含量的检测。药代参数提示EPR在鸡体内的吸收代谢迅速，可较快消除。

Abstract:

The aim of the study was to establish a high performance liquid chromatography (HPLC) method for the determination of eprinomectin (EPR) in chicken plasma, and to evaluate the pharmacokinetics of EPR in chicken. EPR in plasma was extracted with methanol and purified by Sep-Pak C18 solid phase extraction. After purification, the EPR was dried, derivated with trifluoroacetic anhydride and N-methylimidazole, which was detected by HPLC with fluorescence detection (HPLC-FLD). EPR in plasma of chicken showed a good linear relationship within the concentration in the range of 0.1~100 ng/mL, and the correlation coefficient was r=0.9999. The limit of detection (LOD) was 0.1 ng/mL, and the limit of quantitation (LOQ) was 0.3 ng/mL in chicken plasma. The intra- or inter-assay recovery rates of EPR were both higher than 90%, and the intra- and inter-assay coefficient of variation were 2.81~8.02% and 4.32~5.83%, respectively. Layer were orally administrated with EPR at a dose of 5.0mg/kg. The pharmacokinetic parameters of EPR in chicken showed that the concentration of EPR in plasma reached peak values (Cmax) of 354.27 ng/mL at 1.58 h (Tmax); the elimination half-life (T1/2el) was 5.52 h; the mean residence time (MRT) was 6.40 h. The above results indicated that the HPLC method had high sensitivity, good accuracy and less interference, which could be used to determine the concentration of EPR in chicken plasma. The pharmacokinetic parameters suggested that EPR was absorbed and distributed rapidly and eliminated quickly in chickens.