衍生化高效液相色谱法测定醋酸西曲瑞克中10种手性氨基酸的含量

Determination of 10 chiral amino acids in cetrorelix acetate by derivatization HPLC method

目的衍生化高效液相色谱法(HPLC)测定醋酸西曲瑞克中10种手性氨基酸的含量。方法醋酸西曲瑞克水解后,采用Marfey法进行柱前衍生化反应,使用Hedera ODS-2 C18(4.6 mm×250 mm,5μm)色谱柱,流动相为0.065%甲酸溶液(A)-乙腈(B),按程序进行梯度洗脱,紫外检测器检测波长为340 nm。结果醋酸西曲瑞克水解后各氨基酸之间分离度良好,LOD和LOQ的浓度均分别为0.68μmol·L^(-1)和2.04μmol·L^(-1),组成醋酸西曲瑞克的10种手性氨基酸线性范围为10.20~24.49μmol·L^(-1)。结论本方法专属性强,准确度高,可用于同时测定醋酸西曲瑞克中10种手性氨基酸的含量。

Objective To determine the content of 10 chiral amino acids in cetrorelix acetate based on the pre-column derivatization HPLC method.Methods After hydrolysis of cetrorelix acetate,Marfey’s method was used for the pre-column derivatization reaction.Hedera ODS-2 C18 column(4.6 mm×250 mm,5μm)was used,with gradient elution of 0.065%formic acid solution and acetonitrile as the mobile phase.The detection wavelength of the ultraviolet detector was 340 nm.Results All the 10 chiral amino acids in cetrorelix acetate were well separated.The concentration of LOD and LOQ were 0.68μmol·L^(-1) and 2.04μmol·L^(-1) respectively.The linear range was 10.20~24.49μmol·L^(-1).Conclusion This method is specific and accurate for the simultaneous content determination of the 10 chiral amino acids in cetrorelix acetate.