Ksenofontov A L, Boyko A I, Mkrtchyan G V, Tashlitsky V N, Timofeeva A V, Graf A V, Bunik V I, Baratova L A
Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow, 119991, Russia.
Biochemistry (Mosc). 2017 Oct;82(10):1183-1192. doi: 10.1134/S000629791710011X.
An optimized method for analysis of free amino acids using a modified lithium-citrate buffer system with a Hitachi L-8800 amino acid analyzer is described. It demonstrates clear advantages over the sodium-citrate buffer system commonly used for the analysis of protein hydrolysates. A sample pretreatment technique for amino acid analysis of brain extracts is also discussed. The focus has been placed on the possibility of quantitative determination of the reduced form of glutathione (GSH) with simultaneous analysis of all other amino acids in brain extracts. The method was validated and calibration coefficient (K) was determined. Examples of chromatographic separation of free amino acids in extracts derived from different parts of the brain are presented.
描述了一种使用改良柠檬酸盐缓冲系统和日立L-8800氨基酸分析仪分析游离氨基酸的优化方法。该方法相较于常用于分析蛋白质水解产物的柠檬酸钠缓冲系统具有明显优势。还讨论了脑提取物氨基酸分析的样品预处理技术。重点在于定量测定还原型谷胱甘肽(GSH)并同时分析脑提取物中所有其他氨基酸的可能性。对该方法进行了验证并确定了校准系数(K)。给出了源自脑不同部位提取物中游离氨基酸的色谱分离示例。