HPLC法对复方安替比林滴耳液中有关物质的测定

Determination of Related Substances in Compound Antipyrine Ear Drops by IIPLC

目的建立复方安替比林滴耳液中的有关物质测定方法。方法Agilent 1260 Infinity高效液相色谱(HPLC)仪,Agilent ZORBAX SB-C_(18)(4.6 mm×250 mm,5μm)色谱柱,以(0.05 mol·L^(-1)KH_(2)PO_(4)-0.01%三氟乙酸-水)-甲醇-乙腈(65:14:21)为流动相,检测波长为210nm,流速为1.0ml·min^(-1),柱温:30℃,进样体积:20μl,洗脱时间:30min。结果复方安替比林滴耳液中的主峰与杂峰以及杂峰与杂峰之间分离度均大于1.5,杂质吡唑酮和2,6-二甲基苯胺的定量限分别为5.0和1.0μg·m1^(-1),且在各自的线性范围内线性关系良好(R^(2)=0.999,n=6),RSD分别为0.6%和0.5%。结论该HPLC测定方法专属性较好,灵敏度较高,能有效地测定复方安替比林滴耳液中的有关物质吡唑酮和2,6-二甲基苯胺。

Objective To establish a method for the determination of related substances in compound antipyrine ear drops.Methods The conditions of chromatography included Agilent ZORBAX SB-C_(18)(4.6 mm×250 mm,5μm)with the Agilent 1260 infinity HPLC,and(0.05 mol·L^(-1)KH_(2)PO_(4)-0.01%trifluoroacetic acid-water)-methanol-acelonitrile at a ratio of 65:14:21 as the mobile phase.The detection wavelength was 210 nm,the flow rate 1.0 ml·min^(-1)and the column temperature was 30℃.The injection volume was 20μl and the elution time was 30 min.Results The resolutions between the main peak and the impurity peak,and those between the impurity peaks in the compound antipyrine ear drops exceeded 1.5.The quantitative limits of impurities of pyrazolone and 2,6-dimethylaniline were 5.0 and 1.0μg·ml^(-1),respectively.The calibration curves of two known impurities were linear in the self-concentration range(R^(2)=0.999,n=6).The RSDs were 0.6%and 0.5%,respectively.Conclusion This method has a good specificity and sensitivity,which can be used for the determination of related substances in compound antipyrine ear drops.