Determination of Related Substances of 2-(4-Nitrophenyl)Butanoic Acid by High Performance Liquid Chromatography
Objective To establish a high performance liquid chromatography(HPLC)method for the detection of related substances in 2-(4-nitrophenyl)butyric acid.Methods Octadecylsilane bonded silica gel was used as filler[Agilent Infinity Lab Poroshell 120 EC-C18(4.6 mm×250 mm,4 μm)was recommended,or chromatographic column with equivalent efficiency],using water(adjusted to pH 2.5 with phosphoric acid)-acetonitrile(90∶10)as mobile phase A and water-acetonitrile(10∶90)as mobile phase B for gradient elution.The flow rate was 1.0 ml/min,the column temperature was 30 ℃,the detection wavelength was set at 215 nm,and the injection volume was 10 μl.Results The linearity of 2-(4-nitrophenyl)butyric acid and its 10 impurities was good within certain linear ranges.The resolution between 2-(4-nitrophenyl)butyric acid and impurity i-8 was greater than 1.2,and the resolution between the peaks of other components was greater than 1.5.The detection limit of each component was 0.02-0.07 μg/ml.The RSDs of the recoveries of all the impurities in the precision test were not higher than 6％.The recoveries of all impurities were within the range of 92.17％-106.40％.There was no significant changes in the contents of the relevant substances within 27.5 h at room temperature.Replacing the chromatographic column,changing the column temperature,flow rate,pH of the mobile phase,and initial ratio of the mobile phase did not result in significant changes in the detection results of the relevant substances.Conclusion This method has strong specificity and high sensitivity,and it can be used for the detection of related substances of 2-(4-nitrophenyl)butyric acid.
万方数据
维普
