Lavi L E, Holcenberg J S, Cole D E, Jolivet J
J Chromatogr. 1986 Apr 25;377:155-63. doi: 10.1016/s0378-4347(00)80770-7.
The analytical methodologies for the determination of free amino acids in plasma, serum, erythrocytes and leukemic cells are described. Deproteinization of the sample by methanol or organic acids is followed by derivatization with phenylisothiocyanate to form stable phenylthiocarbamylamino acid derivatives. The derivatives are separated by reversed-phase high-performance liquid chromatography in 80 min using a 5-microns C18 column (250 X 4 mm I.D.) and monitored by ultraviolet detection at 254 nm. Twenty physiological amino acids are resolved and quantified in plasma and erythrocyte samples. The resolution and sensitivity of the analytical method permitted unequivocal quantification of very low asparagine and glutamine levels in leukemic cells and growth media following treatment with asparaginase and glutaminase enzymes despite the presence of high aspartic and glutamic acid levels.
本文描述了测定血浆、血清、红细胞和白血病细胞中游离氨基酸的分析方法。用甲醇或有机酸对样品进行脱蛋白处理，随后用异硫氰酸苯酯进行衍生化反应，以形成稳定的苯硫代氨基甲酰氨基酸衍生物。使用5微米的C18柱（内径250×4毫米），通过反相高效液相色谱在80分钟内分离这些衍生物，并在254纳米处通过紫外检测进行监测。在血浆和红细胞样品中可分离并定量20种生理性氨基酸。尽管存在高浓度的天冬氨酸和谷氨酸，但该分析方法的分辨率和灵敏度能够明确地定量白血病细胞和生长培养基在用天冬酰胺酶和谷氨酰胺酶处理后极低水平的天冬酰胺和谷氨酰胺。
