高效液相色谱法测定阿司匹林丁香酚酯咀嚼片的有关物质

doi:10.16431/j.cnki.1671-7236.2023.07.040

摘要/Abstract

摘要： 【目的】采用高效液相色谱（high performance liquid chromatography，HPLC）法测定阿司匹林丁香酚酯（aspirin eugenol ester，AEE）咀嚼片的有关物质，建立一种更简便、准确的检测方法，应用于AEE咀嚼片的质量控制研究。【方法】通过HPLC法考察方法专属性、线性范围、检测限与定量限、精密度、重复性、稳定性、回收率、破坏性试验、耐用性试验及样品测定，综合评价AEE咀嚼片有关物质测定方法，并以自身对照法计算AEE咀嚼片有关物质的量。选用Phenomenex Luna C18（150 mm×4.6 mm，5 μm）色谱柱，柱温为35 ℃；流动相以0.5%磷酸水溶液为A相，以乙腈为B相，梯度洗脱；流速为1.0 mL/min；检测波长为279 nm；进样量为10 μL。【结果】AEE咀嚼片有关物质的HPLC测定方法专属性高，被检测峰与其他峰分离度良好；AEE在0.26~26.00 μg/mL浓度范围内与峰面积线性关系良好，R²=1（n＝7）；有关物质杂质A在0.25~25.50 μg/mL浓度范围内与峰面积线性关系良好，R²＝0.9996（n＝7）；AEE与杂质A定量限分别为0.52及0.51 μg/mL，检测限分别为0.26及0.25 μg/mL；精密度相对标准偏差（RSD）<1%；日内重复性与日间重复性的RSD分别为0.88%及2.50%，稳定性的RSD为1.84%；平均回收率为100.14%（n＝9，RSD=1.66%）。破坏性试验结果显示，不同破坏条件下物料基本守恒，物料守恒均在95%~105%之间。耐用性试验表明，该检测条件的耐用性良好，RSD值为1.87%。3批样品中，AEE咀嚼片杂质A含量均<0.5%，总杂质含量均<1.0%。【结论】该方法操作简便、专属性强、准确度高，可用于AEE咀嚼片中有关物质测定。

关键词: 阿司匹林丁香酚酯(AEE); 咀嚼片; 高效液相色谱; 有关物质

Abstract: 【Objective】 The study was to determine the related substances of aspirin eugenol ester (AEE) chewable tablets by high performance liquid chromatography (HPLC), and establish a more convenient and accurate detection method for the quality control of AEE chewable tablets.【Method】 The specificity, linear range, limits of detection and quantification, precision, repeatability, stability, recovery, destructive testing, durability testing, and sample determination were investigated by HPLC, in order to comprehensively evaluate the determination method of related substances in AEE chewable tablets, and calculate the content of related substances by self-contrast method.The chromatographic separation was performed using a Phenomenex Luna C18 column (150 mm×4.6 mm, 5 μm) at 35 ℃.The mobile phase consisted of water containing 0.5% phosphoric acid (A) and acetonitrile (B) with gradient elution.The flow rate was 1.0 mL/min, the detection wavelength was 279 nm, and the injection volume was 10 μL.【Result】 The results showed that the HPLC method of AEE chewable tablets had high specificity, and the detected peaks were well separated from other peaks.The linear relation between peak area and concentration of the AEE was good in the range of 0.26-26.00 μg/mL, R²=1 (n=7).In addition, the linear relation between peak area and concentration of the impurity A was well in the range of 0.25-25.50 μg/mL, R²=0.9996 (n=7).The limits of quantification of AEE and impurity A were 0.52 and 0.51 μg/mL, and the limits of detection were 0.26 and 0.25 μg/mL, respectively.The relative standard deviation (RSD) of precision was less than 1%.RSD of intraday repeatability, daytime repeatability and stability were 0.88%, 2.50% and 1.84%, respectively.Average recovery was 100.14% (n=9, RSD=1.66%).Destructive test showed that material was basically conserved under different destruction conditions, and the material conservation was between 95% and 105%.The durability test showed that the durability of the detection conditions was good, RSD was 1.87%.In the three batches of samples, the content of impurity A in AEE chewable tablets was less than 0.5%, and the total impurity content was less than 1.0%.【Conclusion】 The method was validated to be simple, specific, accurate and suitable for the determination of related substances in AEE chewable tablets.

Key words: aspirin eugenol ester (AEE); chewable tablets; high performance liquid chromatography; related substances

中图分类号:

S859.79

白莉霞, 刘希望, 杨亚军, 秦哲, 李世宏, 葛闻博, 李剑勇. 高效液相色谱法测定阿司匹林丁香酚酯咀嚼片的有关物质[J]. 中国畜牧兽医, 2023, 50(7): 3007-3016.

BAI Lixia, LIU Xiwang, YANG Yajun, QIN Zhe, LI Shihong, GE Wenbo, LI Jianyong. Determination of Related Substances in Aspirin Eugenol Ester Chewable Tablets by High Performance Liquid Chromatography[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(7): 3007-3016.

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