高效液相色谱法同时测定复方硫酸新霉素滴眼液中7种抑菌剂

Simultaneous Determination of Seven Bacteriostatic Agents in Compound Neomycin Sulfate Eye Drops by HPLC

目的建立同时测定复方硫酸新霉素滴眼液中羟苯甲酯、羟苯乙酯、羟苯丙酯、苯甲酸钠、硫柳汞、苯扎氯铵、苯扎溴铵7种抑菌剂含量的高效液相色谱(HPLC)法。方法色谱柱为月旭Ultimate®XB-C_(18)柱(150 mm×4.6 mm,5μm),流动相为甲醇-磷酸水溶液(pH=3.5),梯度洗脱,流速为1.0 mL/min,柱温为30℃,检测波长为214 nm,进样量为10μL。结果羟苯甲酯、羟苯乙酯、羟苯丙酯、苯甲酸钠、硫柳汞、苯扎氯铵、苯扎溴铵的质量浓度分别在0.625~62.500μg/mL、1.080~108.000μg/mL、0.535~53.500μg/mL、1.015~101.500μg/mL、1.210~121.00μg/mL、4.965~496.500μg/mL、5.350~535.000μg/mL范围内与峰面积线性关系良好(r=0.9999,n=7);精密度、稳定性、重复性试验的RSD均低于1.50%;加样回收率分别为97.24%,98.67%,99.85%,98.55%,97.99%,98.72%,97.55%,RSD分别为0.93%,1.02%,0.97%,0.87%,0.67%,0.85%,0.88%(n=9)。结论该方法操作简便,结果准确,稳定性好,能同时测定复方硫酸新霉素滴眼液中7种抑菌剂的含量。

Objective To establish a high-performance liquid chromatography(HPLC)method for the simultaneous determination of seven bacteriostatic agents such as methylparaben,ethylparaben,propylparaben,sodium benzoate,thiomersal,benzalkonium chloride and benzalkonium bromide in Compound Neomycin Sulfate Eye Drops.Methods The chromatographic column was Welch Ultimate®XB-C_(18)column(150 mm×4.6 mm,5μm),the mobile phase was methanol-phosphoric acid aqueous solution(pH=3.5),the elution method was gradient elution,and the flow rate was 1.0 mL/min,the column temperature was 30℃,the detection wavelength was 214 nm,and the injection volume was 10μL.Results The linear ranges of methylparaben,ethylparaben,propylparaben,sodium benzoate,thiomersal,benzalkonium chloride and benzalkonium bromide were 0.625-62.500μg/mL,1.080-108.000μg/mL,0.535-53.500μg/mL,1.015-101.500μg/mL,1.210-121.00μg/mL,4.965-496.500μg/mL and 5.350-535.000μg/mL(r=0.9999,n=7),the RSDs of precision,stability and repeatability tests were lower than 1.5%,and the recovery rates were 97.24%,98.67%,99.85%,98.55%,97.99%,98.72%and 97.55%with the RSDs of 0.93%,1.02%,0.97%,0.87%,0.67%,0.85%and 0.88%(n=9).Conclusion The method is simple,accurate and stable,which can be used for the simultaneous determination of seven bacteriostatic agents in Compound Neomycin Sulfate Eye Drops.