用RP-HPLC法测定溴芬酸钠滴眼液中溴芬酸钠及有关物质的含量

Content determination of bromfenac sodium and related substances in bromfenac sodium eye drops by RP-HPLC method

目的:建立测定溴芬酸钠滴眼液中溴芬酸钠及有关物质含量的RP-HPLC法。方法:采用Capcellpak C18色谱柱(150mm×4.6mm,5μm),流动相:0.05mol/L乙酸铵(用冰乙酸调节pH至6.0)-甲醇-四氢呋喃(68∶22∶10,V/V/V),流速1.0ml/min,柱温40℃,检测波长270nm。结果:在建立的色谱条件下,有关物质的峰与主峰完全分离,且专属性良好。有关物质的检出限为1.22×10-5 mg/ml,线性范围为0.051～10.07μg/ml,相关系数(r)=0.999 7。溴芬酸钠含量测定的线性范围为0.012 6～0.302 0mg/ml,r=0.999 9,精密度良好。低、中、高浓度(0.16、0.20、0.24mg/ml)的加样回收率分别为(100.7±0.6)%、(100.2±1.1)%和(100.2±0.7)%(n=3)。结论:本研究建立的RP-HPLC法适用于溴芬酸钠滴眼液中溴芬酸钠及有关物质的含量测定。

Objective:To establish a RP-HPLC method for content determination of bromfenac sodium and related sub-stances in bromfenac sodium eye drops.Methods:Capcellpak C18column(150mm×4.6mm,5μm)was performed with the mo-bile phase consisting of0.05mol/L ammonium acetate(adjusting pH to6.0by glacial acetic acid)-methanol-tetrahydrofuran(68:22:10,V/V/V)at a flow rate of1.0ml/min.The column temperature was set at40℃and the determination wave-length was270nm.Results:The peaks of the related substances were well separated from the peak of bromfenac sodium,with good specificity.The detection limit of related substances was1.22×10^-5mg/ml.The concentrations of related substances were in good linearity with the peak area within the ranges of0.051-10.07μg/ml(r=0.9997).The concentration of bromfenac sodi-um was in good linearity with the peak area within the ranges of0.0126-0.3020mg/ml(r=0.9999),with good precision.The recovery rates of low,middle and high concentrations(0.16,0.20,0.24mg/ml)were(100.7±0.6)%,(100.2±1.1)%and(100.2±0.7)%(n=3),respectively.Conclusion:The established RP-HPLC method could be used for content determina-tion of bromfenac sodium and related substances in bromfenac sodium eye drops.