HPLC法测定氨酚曲马多片有关物质方法的建立

Determination of the Related Substances in Paracetamol and Tramadol Hydrochloride Tablets by HPLC

目的:建立HPLC法测定氨酚曲马多片中的有关物质。方法:色谱柱:Agilent ZORBAX SB Phenyl(250 mm×4.6 mm,5μm);流动相:四氢呋喃-水-三氟乙酸-三乙胺(5∶95∶0.1∶0.1)(pH2.2~2.4);流速:1.0 ml·min^(-1);柱温:50℃;检测波长:216 nm;进样体积:30μl。测定6个厂家共16批样品的有关物质含量。结果:在该色谱条件下,两主成分峰及其相应的杂质峰均能有效分离,辅料不干扰有关物质的测定;曲马多和两个已知杂质O-去甲曲马多和顺式曲马多(曲马多杂质A)分别在0.078~15.58(r=1.0000),0.077~15.25(r=0.9999),0.077~15.35μg·ml^(-1)(r=0.9999)的范围内线性关系良好;定量限分别为0.16,0.10,0.13μg·ml^(-1);检测限分别为0.047,0.029,0.038μg·ml^(-1)。结论:建立的方法专属性强,准确度高,重复性好,优于现行标准方法,能较好的区分不同企业氨酚曲马多片的质量,可用于氨酚曲马多片有关物质的质量控制。

Objective:To develop an HPLC method for the determination of the related substances in paracetamol and tramadol hydrochloride tablets.Methods:The chromatography separation was performed on an Agilent ZORBAX SB Phenyl(250 mm×4.6 mm,5μm)column with tetrahydrofuran-water-trifluoroacetic-triethylamineacid(5∶95∶0.1∶0.1)(pH2.2-2.4)as the mobile phase.The flow rate was 1.0 ml·min^(-1),the column temperature was 50℃,the detection wavelength was 260 nm,and the volume injection was 30μl.The contents of related substances in 16 batches of samples from 6 manufacturers were determined.Results:Under the chromatographic conditions,the two principal component peaks and the corresponding impurity peaks could be separated effectively,and the excipients showed no interference with the determination of related substances.Tramadol and two known impurities,O-demethyl-tramadol and cis-tramadol(tramadol related compound A)showed good linearity within the concentration range of 0.078-15.58μg·ml^(-1)(r=1.0000),0.077-15.25μg·ml^(-1)(r=0.9999)and 0.077-15.35μg·ml^(-1)(r=0.9999),respectively.The limits of quantification were 0.16μg·ml^(-1),0.10μg·ml^(-1) and 0.13μg·ml^(-1),and the limits of detection were 0.047μg·ml^(-1),0.029μg·ml^(-1) and 0.038μg·ml^(-1),respectively.Conclusion:The method is specific and accurate with good reproducibility,which is better than the existing standard method,and can be used for the determination of related substances in paracetamol and tramadol hydrochloride tablets,and for distinguishing the quality of products from different enterprises.