Clinimix E(克灵麦)中色氨酸杂质M的测定

Determination of Tryptophan Impurity M Content in Clinimix E

目的:建立Clinimix E(克灵麦)中氨基酸注射液色氨酸杂质M的测定方法。方法:采用HPLC法测定,以十八烷基硅烷键合硅胶为填充剂;以磷酸二氢钠缓冲液(取磷酸二氢钠约3.9 g,加水溶解至1000 mL,加2.9 g·L^(-1)磷酸溶液1000 mL,以磷酸调节pH值至2.3)-乙腈=88.5∶11.5为流动相A,以磷酸二氢钠缓冲液(pH2.3)-乙腈=65∶35为流动相B,采用梯度洗脱,流速为每分钟0.7 mL;检测波长为220 nm;柱温为40℃。结果:在0.2028~4.0560μg·mL^(-1)范围内,色氨酸杂质M的浓度与峰面积之间线性关系良好;该方法的重复性和准确度的RSD值分别为0%、0.2%。结论:该方法专属性、重现性、准确度良好,可用于Clinimix E(克灵麦)中氨基酸腔袋色氨酸杂质M的测定。

A method for determination of tryptophan impurity M in amino acid injectionof Clinimix E.was established.HPLC was used to determine,octadecylsilane silica gel was used as filler,sodium dihydrogen phosphate buffer(pH=2.3)-acetonitrile(volume ratio 88.5∶11.5)was used as mobile phase A,sodium dihydrogen phosphate buffer(pH=2.3)-acetonitrile(volume ratio 65∶35)was used as mobile phase B,gradient elution was adopted,the flow rate was 0.7mL·min^(-1),the detection wavelength was 220 nm,the column temperature was 40℃.The results showed that,there was good linear relationship between the mass concentration of tryptophan impurity M and the peak area method at the range of 0.2028~4.0560μg·mL^(-1).The RSD values for the repeatability and accuracy of the method were 0 and 0.2%,respectively.The method has good specificity,reproducibility and accuracy,and can be used for the determination of tryptophan impurity M content in Clinimix E.