目的 建立快速高通量测定滴眼液中10个抑菌剂(氯己定、苯甲醇、三氯叔丁醇、山梨酸、苯甲酸、对羟基苯甲酸甲酯、对羟基苯甲酸乙酯、对羟基苯甲酸丙酯、硫柳汞和苯扎氯铵)的高效液相色谱方法并探索其抑菌效力。方法 色谱柱为Waters XBridge C18 (4.6 mm×150 mm,5 μm);流动相为0.1%磷酸溶液-甲醇-四氢呋喃;梯度洗脱,流速为1.0 mL·min-1;检测波长为214 nm;对两种滴眼液的抑菌效力进行评价。 结果 10个抑菌剂的峰面积与浓度线性关系良好(r为0.999 3~1.000 0),加样回收率为96.1%~101.8%。一种滴眼液抑菌效力不符合规定。结论 建立的高通量测定方法简便快速,准确,重复性好,样品基质等几乎不干扰测定,可用于滴眼液中抑菌剂的快速高通量测定。研发过程中应对抑菌剂的添加剂量进行处方筛选或进行最低抑菌效力考察代替经验值。
Abstract
OBJECTIVE To establish an HPLC method for rapid screening and simultaneous determination of 10 preservatives in eye drops and assess their antimicrobial effectiveness. METHODS A Waters XBridge C18 column (4.6 mm×150 mm,5 μm) was used, and the mobile phase was 0.1% phosphoric acid-methanol-THF eluted in gradient mode at a flow rate of 1.0 mL·min-1. The detection wavelength was set at 214 nm. The antimicrobial effectiveness of the preservatives was assessed according to Ch. P 2015. RESULTS The calibration curves were linear in the range of the corresponding test concentrations (r=0.999 3-1.000 0). The recoveries were between 96.1%-101.8%. One of the eye drops products did not meet the requirement of Ch. P 2015. CONCLUSION The established method is rapid and inexpensive. And it ensures excellent simplicity, sensitivity, specificity, and reproducibility and can be used for rapid screening and determination of the contents of preservatives in eye drops. The amount of preservatives should be established during the R&D period or determined according to the RESULTS of antimicrobial effectiveness test rather than using empirical values.
关键词
滴眼液 /
抑菌剂 /
对羟基苯甲酸乙酯 /
苯扎氯铵 /
高效液相色谱法 /
抑菌效力
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Key words
eye drops /
preservatives /
ethyl paraben /
benzalkonium chloride /
HPLC /
antimicrobial effectiveness
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中图分类号：
R917
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参考文献
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