反相高效液相色谱法测定甲氧普烯原料中有关杂质的含量OA

A reversed-phase high performance liquid chromatography(RP-HPLC)method was established for determination of the impurities in methoprene raw materials.The analysis was performed on an Agilent Zorbax SB-C18 column(250×4.6 mm,5 μm).The gradient mobile phase was using acetonitrile-methanol-1%glacial acetic acid solution(V/V,47∶21∶32)as mobile phase A,acetonitrile as mobile phase B.The detection wavelength was 264 nm,the flow rate was 1.0 mL/min,the column temperature was 30℃,and the injection volume was 20 μL.The relative retention time was used to determine the position of the known impurities,the correction factor was used to determine the content of related substances in methoprene.Under the proposed chromatographic conditions,the chromatographic peak of methoprene and the related substances were separated completely,and the diluent didn't interfere in the determination of methoprene and the related substances.The relative retention time of impurity B,C,D,A was 0.50,0.64,0.67 and 0.96 respectively.The relative correction factors of impurity B,C,D,A were measured by standard curve method as 0.95,0.92,1.10 and 1.17.Methoprene has a good linear relationship in the range of 0.0261 μg/mL～0.3908 μg/mL(r=1.0000);the four impurities have a good linear relationship within the corresponding concentration range(r＞0.9990).The precision,reproducibility,stability and specificity tests of methoprene and four known impurities were good.The average recoveries of four known impurities were between 100.2%～102.8%.The method is simple and accurate with high specificity,which can be used to determine the impurities of methoprene.