高效液相色谱法同时测定盐酸丁卡因滴眼液中盐酸丁卡因和羟苯乙酯的含量

Determination of tetracaine hydrochloride and ethyl hydrobenzoate in tetracaine hydrochloride eye drops by HPLC

目的:建立高效液相色谱法(high-performanceliquidchromatography,HPLC)测定盐酸丁卡因滴眼液中盐酸丁卡因和羟苯乙酯的含量。方法:采用Agilent Eclipse PLUS C18色谱柱(250 mm×4.6m m, 5μm);以1%三乙胺溶液(三乙胺1 0m L,加水9 9 0m L,用冰醋酸调节p H值至5.0±0.5)-乙腈(65:35)为流动相,流速为1.0m L/min;柱温为30℃;检测波长为256nm;进样体积2 0μL。结果:盐酸丁卡因在0.0 5~0.3 6m g/m L范围内线性关系良好(r=1.0 0 0),平均回收率为99.2%,相对标准偏差(relative standard deviation,R SD)为0.3%(n=9),羟苯乙酯在3.02~24.14μg/m L范围内线性关系良好(r=1.000),平均回收率为98.2%,RSD为0.4%(n=9),该方法重现性好。对照品溶液和供试品溶液在室温放置24h基本稳定;结论:本方法简便、快速、准确。适用于检测盐酸丁卡因滴眼液中盐酸丁卡因和羟苯乙酯的含量。

Objective:To establish a high-performance liquid chromatography(HPLC) method for the determination of tetracaine hydrochloride and ethyl hydrobenzoate in tetracaine hydrochloride eye drops.Methods:The analytic column was Agilent Eclipse Plus C18(4.6 mm ×250 mm,5 μm) and the mobile phase was 1% triethylamine solution(10 mL triethylamine and 990 mL water,pH adjusted to 5.0±0.5 with glacial acetic acid)-acetonitrile(65:35) at a flow rate of 1.0 mL/min.The detection wavelength was 256 nm and the column temperature was30℃.The injection volume was 20 μL.Results: The linear range of tetracaine hydrochloride was well shown within 0.05-0.36 mg/mL(r=1.000).The average recovery rate of tetracaine hydrochloride was 99.2% with relative standard deviation(RSD) 0.3%(n=9).The linear range of ethyl hydrobenzoate was well shown within3.02-24.14 μg/mL(r=1.000).The average recovery rate of tetracaine hydrochloride was 98.2% with RSD 0.4%(n=9).The method had high reproducibility.The reference solution and testing solution were stable for 24 h in room.Conclusion:The method is simple,quick and accurate,which is suitable for the determination of tetracaine hydrochloride and ethyl hydrobenzoate in tetracaine hydrochloride eye drops.