Content Determination of Related Substances in Fentanyl Citrate Injection by HPLC
Objective To establish a high-performance liquid chromatography(HPLC)method for the content determination of related substances in Fentanyl Citrate Injection.Methods The chromatographic column was Shim-pack scepter C18-120 column(150 mm×4.6 mm,5 µm),the mobile phase was 0.01 mol/L ammonium chloride solution-acetonitrile(950∶50,V/V)-0.01 mol/L ammonium chloride solution-acetonitrile(400∶600,V/V)with gradient elution,the flow rate was 1.0 mL/min,the detection wavelength was 210 nm,the column temperature was 40℃,and the injection volume was 100 µL.Results The linear ranges of impurity Ⅰ,impurity Ⅱ,impurity Ⅲ and impurity Ⅺ were 0.02-0.50 µg/mL,0.02-0.50 µg/mL,0.02-0.49 µg/mL,and 0.02-0.50 µg/mL(r>0.993),respectively.The limits of detection(LOD)were 1.00,1.00,0.98,and 1.00 ng,respectively.The limits of quantitation(LOQ)were 2.01,2.00,1.97,and 2.01 ng,respectively.The RSD of each impurity in the repeatability test was lower than 2.0%(n = 6).The RSD of each impurity in the intermediate precision test was lower than 3.0%(n = 12).The average recoveries of impurity Ⅰ,impurity Ⅱ,impurity Ⅲ and impurity Ⅺ were 98.03%,100.04%,97.94%,and 101.03%,with RSDs of 1.61%,1.09%,0.98%,and 1.36%(n = 9),respectively.The content of specific and unknown impurities in the pilot samples of two specifications(three batches each)was lower than 0.05%(disregard limit).Conclusion The method is simple,specific,and sensitive,with good durability,high accuracy and precision,the separation of various impurities in this method is superior to the methods in the Chinese Pharmacopoeia and the British Pharmacopoeia,it is suitable for the determination of related substances in Fentanyl Citrate Injection.The impurity Ⅰ,impurity Ⅱ,impurity Ⅲ,and impurity Ⅺ in the samples shall not exceed 0.5%of the labeled amount,the maximum unknown impurities shall not exceed 0.2%,and the total unknown impurities shall not exceed 0.75%.
万方数据
维普
