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目的建立茚三酮柱后衍生化氨基酸分析仪测定N(2)-L-丙氨酰-L-谷氨酰胺(L-Ala-L-Glu)含量的方法。方法根据L-Ala-L-Glu与茚三酮反应后溶液在570 nm处有最大吸收的原理,以钾钠缓冲液为流动相,将L-Ala-L-Glu洗脱后,与茚三酮进行柱后衍生反应,生成可以被分光光度计检测的有色物质,氨基酸分析仪测定L-Ala-L-Glu的含量。结果 L-Ala-L-Glu在1～20 mg.L-1内线性关系良好(r=0.997 9),低、中、高3种浓度的回收率分别为94.5%,97.6%和98.7%;RSD分别为0.52%,0.10%和0.06%。结论本法操作简单,分离度良好,结果准确可靠,可用于L-Ala-L-Glu的含量测定。
Objective To establish a method for the determination of N (2) -L-alanyl-L-glutamine (L-Ala-L-Glu) by column derivatization with ninhydrin. Methods Based on the principle that L-Ala-L-Glu and ninhydrin reacted at 570 nm, the maximum absorbance was obtained. After the L-Ala-L-Glu was eluted with potassium-sodium buffer as eluent, Ketones for post-column derivatization to produce colored substances which can be detected by spectrophotometer. The content of L-Ala-L-Glu was determined by amino acid analyzer. Results The linearity of L-Ala-L-Glu in 1 ~ 20 mg.L-1 was good (r = 0.997 9). The recoveries of L-Ala-L-Glu were 94.5%, 97.6% and 98.7% RSD were 0.52%, 0.10% and 0.06% respectively. Conclusion This method is simple, good resolution, the results are accurate and reliable, can be used for the determination of L-Ala-L-Glu.