UPLC-UV法同时测定阿莫西林克拉维酸钾片中阿莫西林和克拉维酸的含量及其在国产片剂溶出曲线测定中的应用

Simultaneous determination of amoxicillin and clavulanate in amoxicillin and clavulanate potassium tablets by UPLC- UV and its application in the dissolution profile study in domestic tablets

目的：建立快速准确同时测定阿莫西林克拉维酸钾片中阿莫西林和克拉维酸含量的UPLC-UV方法，并将其应用于阿莫西林克拉维酸钾片溶出曲线的测定，比较不同生产厂家产品在水中的溶出行为。方法：采用Agilent Zorbax Eclipse Plus C18 Rapid Resolution RRHD 色谱柱（2.1 mm×50 mm，1.8 μm），流动相：0.05 mol·L-1磷酸二氢钠溶液（取磷酸二氢钠7.8 g，加水900 mL使溶解，用10%磷酸溶液或氢氧化钠试液调节pH至4.4±0.1，加水稀释至1000 mL）-甲醇（94：6）；流速：0.2 mL·min-1；柱温：35 ℃；检测波长：220 nm。采用中国药典2010年版二部附录X C第二法的装置，分别测定了9个生产企业阿莫西林克拉维酸钾片在水中的溶出曲线，并用AV因子法进行了比较分析。结果：2个组分在1min内全部出峰且分离良好，阿莫西林和克拉维酸进样量分别在19.79~494.85 ng和9.85~246.35 ng范围内均呈良好的线性关系（r=1），平均回收率分别为100.1%和100.0%（n=9），重复性试验测得的精密度分别为0.31%和0.22%（n=6）；定量限分别为1.5 ng和1 ng。AV因子计算结果表明，9个生产企业中只有两家2个组分的的溶出曲线完全相似，其余各企业产品2个组分溶出曲线有差异，差异均具有统计学意义。结论：方法快速简便、准确可靠、灵敏度高、重复性好，满足了快速测定的要求；不同生产企业的辅料、生产工艺和生产规模导致了国产阿莫西林克拉维酸钾片的溶出曲线存在较大的差异。

Objective： To establish a UPLC-UV method for the simultaneous determination of amoxicillin and clavulanate in oral tablets and to investigate the dissolution behavior of the tablets. Methods： The Agilent Zorbax Eclipse Plus C18 Rapid Resolution RRHD Column（2.1 mm×50 mm,1.8 μm）was adopted with the mobile phase consisting of 0.05 mol·L-1 solium dihydrogen phosphate（adjusted to pH 4.4 with 10% phosphoric acid）- methanol（94：6）at the flow fate of 0.2 mL·min-1;the detection wavelength was 220 nm,and the column temperature was at 35 ℃.The dissolution profiles of amoxicillin and clavulanate potassium tablets from 9 pharmaceutical manufacturers（which were represented by A,B,C,D,E,F,G,H,I,respectively）in water were determined by the method 2 in appendix X C of Chinese pharmacopoeia Vol Ⅱ（2010 edition）,and comparatively analyzed by the AV factor method. Results： The two main peaks were well separated in 1 minute and the linear ranges of amoxicillin and clavulanate were 19.79-494.85 ng（r=1）and 9.85 -246.35 ng（r=1）,respectively,the average recoveries were 100.1% and 100.0%（n=9）,respectively,RSDs of repetitive test were 0.31% and 0.22%（n=6）,respectively,the limits of quantization were 1.5 ng and 1.0 ng,respectively.The AV factors among amoxicillin and clavulanate potassium tablets were significantly different.The dissolution profiles of amoxicillin and clavulanate potassium tablets showed significant difference from each other except manufacturer A and B. Conclusion： The established method is sensitive,accurate and specific,which can satisfy the requirement of fast-analysis by shortening the time for an analysis program within 1 minute.The quality of excipients,the formulation and the process and the scale lead to the difference of the dissolution of domestic amoxicillin and clavulanate potassium tablets from different manufacturers.