高效液相法测定5-氟-1H-吲哚-2,3-二酮中基因毒性杂质盐酸羟胺的含量

Determination of Hydroxylamine Hydrochloridein 5- fluoro-1H-indole-2,3-dione by HPLC

[目的]:采用高效液相色谱法检测5-氟-1H-吲哚-2,3-二酮中的基因毒性杂质盐酸羟胺的含量。[方法]:以YMC-PacK ODS-A(4.6mm×250mm,5μm)为色谱柱;流动相为10mmol·L^(-1)磷酸二氢钾缓冲液(用磷酸调节pH值为2.3)-乙腈(70:30);流速为1.0mL·min^(-1);检测波长为254nm;柱温为40℃。[结果]:盐酸羟胺浓度在0.055~0.44μg·min-1范围内与峰面积线性关系良好,线性方程为y=91894x+531.53(r=0.9991);定量下限为0.03μg·mL^(-1);平均回收率为99.9%(RSD=0.9%)。[结论]:本方法适用于5-氟-1H-吲哚-2,3-二酮中盐酸羟胺的检测。

[Objective]:The genotoxic impurity hydroxylamine hydrochloride in 5-fluoro-1H-indole-2,3-dione was determined by HPLC.[Methods]:The chromatographic column was YMC-PacK ODS-A(4.6mm×250mm,5μm).The mobile phase consisted of 10mmol·L^(-1) potassiumdihydrogen phosphate buffer(ph adjusted to 2.3 with phosphoric acid)-acetonitrile(70:30)at a flow rate of 1.0mL·min^(-1).The detection wavelength was 254nm and the column temperature was 40℃.[Results]:The concentration of 5-fluoro-1H-indole-2,3-dione in the range of 0.055~0.44μg·mL^(-1) was in good linear relationship with the peak area.The regression equation was y=91894x+531.53(r=0.9991).The quantification limits of was 0.03μg·mL^(-1).The average recoveries were 99.9%(RSD=0.9%).[Conclusion]:This method is suitable for the determination of hydroxylamine hydrochloride in 5-fluoro-1H-indole-2,3-dione.