乙酰氨基阿维菌素缓释注射剂中乙酰氨基阿维菌素含量与有关物质HPLC检测方法的建立

试验旨在建立一种高效液相色谱检测方法,同时测定乙酰氨基阿维菌素(EPR)缓释注射剂的主成分与有关物质,采用外标法计算主成分的含量,采用自身对照法计算有关物质的含量。色谱柱为Phenomenex Luna 5 μ-C8-100A 250 mm×4.60 mm,5 μm;色谱条件为:流动相为乙腈-0.1%高氯酸水溶液,梯度洗脱,流速为1.5 mL/min,检测波长为245 nm,柱温为35 ℃,进样量为20 μL。结果表明,EPR缓释注射剂主成分与杂质分离度良好,B1a与B1b之间分离度>3;在EPR浓度为1.5625~1 000 μg/mL的范围内线性关系良好;EPR的检测限与定量限分别为0.01和0.34 μg/mL,3种精密度的相对标准偏差(RSD)均<2%,稳定性的RSD为0.28%,加样回收率为102.54%,且RSD为3.14%(n=9)。耐用性试验表明,EPR缓释注射剂主成分的分离度均>1.5,理论塔板数>4 500,表明该检测条件的耐用性良好。3批样品中,EPR缓释注射剂含量的平均值为102.60%,B1b占B1a与B1b之和的1.68%,总杂质的平均含量为2.57%。建立的EPR缓释注射剂中主成分含量与有关物质的测定方法具有简便、专属性强、灵敏度高、结果准确等特点,可用于EPR缓释注射剂的质量控制。

Establishment of HPLC Method for Determination of Eprinomectin and Related Substances in Eprinomectin Sustained-release Injection

The study was designed to develop an HPLC method for the determination of the content of the principal ingredient and related substances in eprinomectin (EPR) sustained-release injection.The external standard method and self-control method were utilized for calculating the content of the principal ingredient and related substances, respectively.The chromatographic column of the content and related substances was Phenomenex Luna 5 μ-C8-100A 250 mm×4.60 mm, 5 μm.The detection method was as followed:Acetonitrile-0.1% aqueous perchloric acid solution with gradient, the flow rate was 1.5 mL/min, the detection wavelength was 245 nm, the column temperature was 35 ℃, and the injection volume was 20 μL.The results showed that the principal component of EPR was well separated with the peak of related substances, the degree of separation between B1a and B1b was greater than 3.The linear relationship was good when the concentration of EPR was 1.5625-1 000 μg/mL.The detection and quantification limit of EPR were 0.01 and 0.34 μg/mL, respectively.RSD of the three kinds of precision was less than 2%, RSD of stability test was 0.28%, average recovery was 102.54%, and RSD was 3.14% (n=9).The robustness showed that the resolution of the principal components of EPR sustained-release injection was more than 1.5, and the number of theoretical plates was more than 4 500.The results showed that the detection conditions were durable.In 3 batches, average value of EPR was 102.60%, B1b accounts for 1.68% of the sum of B1a and B1b, total impurity were 2.57%.All the results suggested that the optimized HPLC method in this study was simple, specific, sensitive, accurate, and suitable for quality control of EPR sustained-release injection.