基于HPLC-MS/MS的2种乙酰氨基阿维菌素注射剂在兔体内的药代动力学对比研究

doi:10.16431/j.cnki.1671-7236.2025.05.033

摘要/Abstract

摘要： 【目的】建立兔血浆中乙酰氨基阿维菌素(EPR)的高效液相色谱-串联质谱(HPLC-MS/MS)检测方法，比较2种EPR注射剂在商品兔体内的药代动力学，为后续制剂开发提供支持。【方法】将12只健康新西兰商品兔，雌雄各半，分为参比制剂(市售EPR注射剂)组和受试制剂(自研EPR缓释注射剂)组，皮下注射相同剂量的2种制剂，于不同时间点采集血液样本，以莫西菌素为内标，采用HPLC-MS/MS法检测EPR的浓度，并对所建立的方法进行特异性、检测限和定量限、标准曲线、准确度和精密度、基质效应、稳定性考察，以软件Phoenix WinNonlin对实际的采样时间-血药浓度数据进行权重为1/Y的非房室分析，求得2种制剂在兔体内的药代动力学参数。【结果】建立的HPLC-MS/MS分析方法检测限和定量限分别为0.5和1 ng/mL；标准曲线在1～200 ng/mL范围内线性关系良好，且相关系数(R²)＞0.99；高、中、低及定量限4个浓度的批内和批间准确度在93.70%～107.73%范围内，批内和批间相对标准差(RSD)均＜15%。参比制剂和受试制剂中EPR达峰时间(T_max)分别为(0.39±0.09)和(0.22±0.04)d；峰浓度(C_max)分别为(112.82±29.40)和(30.86±5.11)ng/mL；半衰期(t_1/2)分别为(7.14±0.58)和(22.15±1.22)d；药时曲线下面积(AUC_0→∞分别为(420.55±21.74)和(300.65±13.62)ng·d/mL；平均滞留时间(MRT)分别为(3.74±0.80)和(41.62±9.04)d。【结论】本研究建立的HPLC-MS/MS分析方法灵敏度和准确度高，稳定可靠，可用于兔血浆中EPR的检测。2种制剂在吸收速率、吸收程度及消除速率方面具有明显差异，与参比制剂相比，受试制剂能显著延长商品兔血浆中EPR存在的时间。

关键词: 乙酰氨基阿维菌素; 高效液相色谱-串联质谱(HPLC-MS/MS); 药代动力学; 缓释注射剂

Abstract: 【Objective】 The purpose of the experiment to develop a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for the determination of eprinomectin (EPR) in rabbit plasma,and to compare the pharmacokinetics of two EPR injections in commercial rabbits,so as to provide support for subsequent formulation development.【Method】 12 healthy New Zealand rabbits,half male and half female,were divided into a reference preparation (marketed EPR injection) group and a test preparation (self-developed EPR extended-release injection) group.After subcutaneously injecting the same dose of the two preparations,the plasma samples were collected at different time points.The concentration of EPR was determined by HPLC-MS/MS using moxidectin as an internal standard.The specificity,detection limit,quantification limit,standard curve,accuracy and precision,matrix effect and stability of the established method were investigated.The actual sampling concentration-time data were analyzed with the software Phoenix WinNonlin for non-compartmental model with a weight of 1/Y,and the pharmacokinetic parameters of the two formulations in rabbits were obtained.【Result】 The limit of detection and the limit of quantitative were 0.5 and 1 ng/mL,respectively.The standard curve had a good linear relationship in the range of 1-200 ng/mL with a correlation coefficient (R²) ＞0.99.The intra-assay and inter-assay accuracies of the four concentrations were in the range of 93.70% to 107.73%,and the intra-assay and inter-assay RSDs were all less than 15%.The results showed that the peak time (T_max) of EPR in the reference preparation and the test preparation was (0.39±0.09) and (0.22±0.04)d respectively,the peak concentrations (C_max) were (112.82±29.40) and (30.86±5.11)ng/mL,the half-lives (t_1/2) was (7.14±0.58) and (22.15±1.22)d respectively,the area under the drug time curves (AUC_0→∞) were (420.55±21.74) and (300.65±13.62) ng·d/mL,the mean retention time (MRT) was (3.74±0.80) and (41.62±9.04)d,respectively.【Conclusion】 The results showed that the established HPLC-MS/MS method was sensitive,accurate,stable and reliable,and could be used for the determination of EPR in rabbit plasma samples.The two preparations differed significantly in the rate and extent of absorption as well as in the rate of elimination.The test formulation significantly prolonged the presence of EPR in rabbit plasma compared to the reference formulation.

Key words: eprinomectin; high-performance liquid chromatography-mass spectrometry (HPLC-MS/MS); pharmacokinetics; extended-release injection

中图分类号:

S859.79⁺5

张贺超, 张振东, 刘希望, 杨亚军, 李世宏, 秦哲, 李剑勇. 基于HPLC-MS/MS的2种乙酰氨基阿维菌素注射剂在兔体内的药代动力学对比研究[J]. 中国畜牧兽医, 2025, 52(5): 2295-2304.

ZHANG Hechao, ZHANG Zhendong, LIU Xiwang, YANG Yajun, LI Shihong, QIN Zhe, LI Jianyong. Comparative Study on Pharmacokinetics of Two Eprinomectin Injections in Rabbits by HPLC-MS/MS[J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(5): 2295-2304.

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