分子排阻色谱-反相高效液相色谱法测定阿莫西林颗粒中高分子聚合物含量

Content Determination of High-Molecular Polymer in Amoxicillin Granules by SEC-RP-HPLC

目的建立测定阿莫西林颗粒中高分子聚合物含量的分子排阻色谱-反相高效液相色谱法。方法色谱柱为Cef-SEC球状亲水硅胶柱(300 mm×7.8 mm,5μm),流动相为磷酸盐缓冲液[pH 7.0,0.005 mol/L磷酸氢二钠-0.005 mol/L磷酸二氢钠(50∶50,V/V)]-乙腈(90∶10,V/V),流速为0.6 mL/min,检测波长为254 nm,柱温为25℃,进样量为20μL。结果阿莫西林质量浓度在2.51~35.07μg/mL范围内与峰面积线性关系良好(r=0.9993,n=8);定量限为5.00 ng,检测限为1.60 ng;精密度、稳定性、重复性、中间精密度、耐用性试验结果均符合要求。国内A公司5批阿莫西林颗粒中高分子聚合物的含量均低于国外L公司的原研阿莫西林细粒。结论该方法专属性强、结果准确、重复性好,可用于测定阿莫西林颗粒中高分子聚合物含量。

Objective To establish a size exclusion chromatography-reversed phase-high performance liquid chromatography(SEC-RP-HPLC)method for the content determination of the high-molecular polymer in Amoxicillin Granules.Methods The chromatographic column was a Cef-SEC spherical hydrophilic silica gel column(300 mm×7.8 mm,5μm),the mobile phase was phosphate buffer solution[pH 7.0,0.005 mol/L disodium hydrogen phosphate-0.005 mol/L sodium dihydrogen phosphate(50∶50,V/V)-acetonitrile(90∶10,V/V)],the flow rate was 0.6 mL/min,the detection wavelength was 254 nm,the column temperature was 25℃,and the injection volume was 20μL.Results The linear range of amoxicillin was 2.51-35.07μg/mL(r=0.9993,n=8).The limit of quantification(LOQ)was 5.00 ng,and the limit of detection(LOD)was 1.60 ng.The results of the precision,stability,repeatability,intermediate precision,and durability tests all met the requirements.The content of high-molecular polymer in the five batches of Amoxicillin Granules produced by the Company A in China was lower than that of original-patented Amoxicillin Granules produced by the Company L in the foreign country.Conclusion This method has strong specificity,accurate results,and good repeatability,which can be used for the content determination of the high-molecular polymer in Amoxicillin Granules.