苯磺贝他斯汀滴眼液中苯扎氯铵含量分析方法研究

Determination of Benzalkonium Chloride in Bepotastine Besilate Eye Drops

目的建立苯磺贝他斯汀滴眼液中抑菌剂苯扎氯铵含量测定的高效液相色谱法(HPLC),并分析滤膜吸附作用对检测结果的影响。方法色谱柱为Agilent TC-C C18(4.6 mm×250 mm,5μm),流动相为乙腈-5 mmol/L乙酸铵溶液(含1%三乙胺,用磷酸调节pH值至5.0)=65:35,检测波长214 nm,进样量20μl;通过控制滤膜材质、来源、过滤体积,考察滤膜吸附差异。结果溶剂与辅料对苯扎氯铵含量测定无干扰,在0.02438~0.07314 mg/ml范围内线性关系良好(y=2.453×107x+2.173×104),仪器精密度RSD为0.44%,重复性RSD为1.25%,中间精密度RSD为1.70%,低、中、高浓度对应的平均回收率为96.94%,RSD为1.28%,24 h内溶液稳定性RSD为0.81%,3批检测含量:112.89%,104.61%,108.22%。滤膜材质、来源、过滤体积均会影响滴眼液中苯扎氯铵吸附情况,不同同系物吸附强度不同,且亲水性滤膜吸附作用相对较小。结论该方法准确可靠,灵敏度较高,适用于对苯磺贝他斯汀滴眼液中苯扎氯铵的检查。

Objective To establish an HPLC method for the determination of the bacteriostatic agent benzalkonium chloride in Bepotastine Besilate Eye Drops, and to analyze the effect of filter adsorption on the detection results. Methods An Agilent TC-C C18(4.6 mm×250 mm, 5 μm) column was used with the mixture of acetonitrile and 5 mmol/L ammonium acetate solution(containing 1 % triethylamine, adjusted to pH 5.0 with phosphoric acid)(65:35) as the mobile phase. The detection wavelength was 214 nm and the injection volume was 20 μl. The filter adsorption difference was investigated by controlling the filter material, source and filtration volume. Results Solvents and excipients did not interfere with the determination of benzalkonium chloride. The linear relationship was good in the range of 0.02438-0.07314 mg/ml(y=2.453×107 x+2.173×104). The precision RSD of the instrument was 0.44 %, the RSD of repeatability was 1.25 %, and the intermediate precision RSD was 1.70 %. The average recovery of low, medium and high concentrations was 96.94 %, with RSD of 1.28 %. The RSD of stability was 0.81 % within 24 h and the contents of 3 batches of preparations were 112.89 %, 104.61 % and 108.22 %, respectively. The material, source and filtration volume of the membrane could all affect the adsorption of benzalkonium chloride in Bepotastine Besilate Eye Drops. The adsorption strength of different homologues was different and the adsorption of hydrophilic membrane was relatively small. Conclusion The method is accurate, reliable and sensitive, and is suitable for the detection of benzalkonium chloride in Bepotastine Besilate Eye Drops.