高效液相色谱法测定更昔洛韦中6种嘌呤类杂质

Determination of six kinds purines impurities in ganciclovir by high performance liquid chromatography

建立高效液相色谱法同时分析更昔洛韦中2-氨基-9-[[(2-氯丙基-2-烯-1-基)氧]甲基]-1,9-二氢-6H-嘌呤-6-酮、(2RS)-2[(2-氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)甲氧基]-3-羟丙基丙酸酯、2-氨基-9-[[(1Rs)-2-氯-1-(羟甲基)乙氧基]甲基]-1,9-二氢-6H-嘌呤-6-酮、2-氨基-9-[[[2-羟基-1-(羟甲基)乙氧基]甲氧基]甲基]-1,9-二氢-6H-嘌呤-6-酮、2-氨基-9-[[(2Rs)-2-3-二羟基丙氧基]甲基]-1,9-二氢-6H-嘌呤-6-酮和2-氨基-1,9-二氢-6H-嘌呤-6-酮6种杂质含量的方法。采用强酸性阳离子交换基团键合硅胶为填充剂的USP L9 SCX型色谱柱(250 mm×4.6 mm,10μm),以0.05%三氟乙酸溶液-乙腈(50∶50)为流动相,柱温为40℃,流量为1.5 mL/min,检测波长为254 nm,进样体积为20μL。结果表明更昔洛韦与相邻杂质峰之间相互的分离度良好。除2-氨基-9-[[[2-羟基-1-(羟甲基)乙氧基]甲氧基]甲基]-1,9-二氢-6H-嘌呤-6-酮外,其它5种杂质分别在各自的质量浓度范围内与色谱峰面积线性关系良好,线性相关系数不小于0.999,方法检出限为4.43~7.25 ng/mL,定量限为14.76~24.16 ng/mL。该方法简单快捷,专属性强,精密度好,可用于更昔洛韦中6种嘌呤类杂质的检测。

A method for the determination of 2-amino-9-[[(2-chloroprop-2-en-1-yl)oxy]methyl]-1,9-dihydro-6H-purin-6-one;(2RS)-2-[(2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl)methoxy]3-hydroxypropyl propionate;2-amino-9-[[(1RS)-2-chloro-1-(hydroxymethyl)ethoxyl]-methyl]-1,9-dihydro-6H-purin-6-one;2-amino-9-[[[2-hydroxy-1-(hydroxymethyl)ethoxyl]-methyl]methyl]-1,9-dihydro-6H-purin-6-one;2-amino-9-[[(2RS)-2,3-dihydroxypropoxy]methyl]-1,9-dihydro-6H-purin-6—one,and 2-amino-1,9-dihydro-6H-purin-one(guanine)in ganciclovir by high performance liquid chromatography was established.A USP L9 SCX column(250 mm×4.6 mm,10μm)was used with strongly acidic cation exchange group bonded silica gel as a filler.0.05%trifluoroacetic acid solution-acetonitrile(50∶50)was used as mobile phase,the column temperature was 40℃,the flow rate was 1.5 mL/min,the detection wavelength was 254 nm,and the injection amount was 20μL.The degree of separation of ganciclovir from adjacent impurity peaks and adjacent impurity peaks was good.Ganciclovir and purines impurities except 2-amino-9-[[[2-hydroxy-1-(hydroxymethyl)ethoxyl]-methyl]methyl]-1,9-dihydro-6H-purin-6-one had a good linear relationship with the chromatographic peak area in their mass concentration range,correlation coefficients were more than 0.999,the detection limit of the method was 4.43-7.25 ng/mL,and the limit of quantification was 14.76-24.16 ng/mL.The method was simple and fast,with strong specificity and good precision,which can be used for the inspection and analysis of 6 kinds purines impurities in ganciclovir.