HPLC法测定美司钠原料中的有关物质

Determination of the Related Substances in Mesna by HPLC

目的:建立HPLC法测定美司钠原料中的有关物质。方法:采用C18柱(250 mm×4.6 mm,5μm);以甲醇-磷酸盐缓冲液(取磷酸二氢钾2.94 g、磷酸氢二钾2.94 g、四丁基硫酸氢铵2.6 g溶于660 ml水中,用磷酸调pH 2.3)(34∶66)为流动相;流速为1.0 ml·min^-1;检测波长为235 nm。结果:美司钠与各杂质及强制破坏产物能完全分离;美司钠及杂质A、B、D、F分别在41.03~4103.00μg·ml^-1(r=1.0000)、0.53~53.44μg·ml^-1(r=1.0000)、0.43~42.88μg·ml^-1(r=1.0000)、30.80~308.00μg·ml^-1(r=1.0000)、0.4224~42.24μg·ml^-1(r=1.0000)范围内线性关系良好;各杂质加样回收率在97.8%~101.4%之间;进样精密度和重复性均符合规定;其中杂质A、B、F的相对校正因子分别为0.03,0.01,0.01。结论:建立的方法可用于美司钠原料有关物质的质量控制。

Objective:To develop an HPLC method for the determination of related substances in mesna.Methods:The analysis was performed on a C18 column(250 mm×4.6 mm,5μm)with the mobile phase of methanol-phosphate buffered solution(2.94 g potassium dihydrogen phosphate,2.94 g dipotassium hydrogen phosphate and 2.6 g tetrabutylammonium hydrogen sulfate were dissolved in ca 660 ml water,and the pH value was adjusted to 2.3 with phosphoric acid)(34∶66)at the flow rate of 1.0 ml·min-1,and the detection wavelength was at 235 nm.Results:Mesna was separated completely from the impurities and degradation products under various conditions.Mesna and impurities A,B,D and F had good linearity within the range of 41.03-4103μg·ml^-1(r=1.0000),0.53-53.44μg·ml^-1(r=1.0000),0.43-42.88μg·ml^-1(r=1.0000),30.80-308.00μg·ml^-1 and 0.42-42.24μg·ml^-1(r=1.0000),respectively.The sampling precision and repeatability both met the requirements.The relative correction factor of impurities A,B and F was 0.03,0.01 and 0.01,respectively.Conclusion:The method is simple,specific and accurate,and can be applied to the quality control of mesna.