HPLC法测定利福布汀原料药及胶囊中有关物质

Determination of Related Substances in Rifabutin Crude Drug and Capsules by HPLC

目的:建立利福布汀原料药及胶囊中有关物质检查的方法。方法:采用高效液相色谱法。色谱柱为Agilent XDB-C8,流动相为乙腈-0.1 mol/L磷酸二氢钾溶液(pH 6.5±0.1)(50:50,V/V),流速为1.0 mL/min,检测波长为254 nm,柱温为30℃,进样量为20μL,供试品以流动相为溶剂制备成质量浓度为1.0 mg/mL的溶液。采用此新建立的方法与利福布汀原料药及胶囊质量标准中的现行方法(供试品溶液质量浓度为0.5 mg/mL,色谱柱为C18)分别进行系统适用性试验,并对利福布汀原料药及胶囊共6批样品进行有关物质检查(峰面积归一化法)。结果:利福布汀的检测质量浓度线性范围为0.8~16μg/mL(r=1.000 0),精密度、重复性和稳定性试验(12 h)的RSD均小于2.0%(n=6),检测限和定量限分别为0.025 4、0.085 2μg/mL。在系统适用性试验中,采用改进方法与现行方法,利福布汀峰与其前降解产物峰的分离度分别为7.50、3.47;在测定6批样品时,采用新方法检出的杂质个数较现行方法多1~5个,杂质总量高出0.19%~0.55%。结论:建立的新方法分离效果好,灵敏度高,可用于利福布汀原料药和胶囊有关物质的检查,更有助于控制药品质量。

OBJECTIVE:To establish a method for the determination of related substances in rifabutin crude drug and capsules by HPLC.METHODS:HPLC method was adopted.The determination was performed on Agilent XDB-C8 column with mobile phase consisted of acetonitrile-0.1 mol/L potassium dihydrogen phosphate solution(pH 6.5±0.1)(50:50,V/V)at the flow rate of1.0 mL/min.The detection wavelength was set at 25/4 nm,the column temperature was 30 ℃,and sample size was 20 μL.The mobile phase was used as solvent to prepare the sample solution with a mass concentration of 1.0 mg/mL.The system suitability test was performed by using newly established method and current method(mass concentration of sample solution 0.5 mg/mL,C18 column)stated in quality standard of rifabutin crude drug and capsules.Related substance test was conducted for 6 batches of rifabutin crude drug and capsule(peak area normalization method).RESULTS:The linear range of rifabutin was 0.8-16μg/mL(r=1.000 0),RSDs of precision,reproducibility and stability tests(12 h)were all lower than 2.0%(n=6);the limits of detection and quantification were 0.025 4,0.085 2 μg/mL.In system suitability test,by using new method and current method,separation degree of rifabutin peak and pre-degradation product peak were 7.50 and 3.47.When 6 batches of samples were determined,the number of impurities detected by this method was 1-5 more than that by the current method,and the total amount of impurities was0.19%-0.55% higher.CONCLUSIONS:Established new method is well-separated and sensitive,and can be used for the determination of related substance in rifabutin crude drug and capsules,which helps the quality control of drugs.