2，4-二硝基氯苯衍生高效液相色谱法测定血清中的谷氨酸和谷氨酰胺  

DETERMINATION OF GLUTAMIC ACID AND GLUTAMINE IN SERUM BY HPLC WITH 2,4-DINITROCHLOROBENZENE DERIVATIZATION

作　　者：骆春迎[1] 黎源倩[1] 邹晓莉[1] 曾红燕[1] 彭海燕[2] 张思维[1] 王习祥[1] 

机构地区：[1]四川大学华西公共卫生学院,成都610041 [2]四川大学华西药学院

出　　处：《现代预防医学》2008年第10期1900-1902,共3页Modern Preventive Medicine

基　　金：四川大学校青年基金资助(校200467)

摘　　要：[目的]建立测定血清中谷氨酸(glutamic acid,Glu)和谷氨酰胺(glutamine,Gln)的高效液相色谱法(high performance liquid chromatography,HPLC)。[方法]血清经三氯乙酸沉淀蛋白,2,4-二硝基氯苯(2,4-dini-trochlorobenzene,DNCB)衍生其中的氨基酸后进行HPLC分离测定。[结果]谷氨酸和谷氨酰胺的检出限分别为0.005μg/ml和0.054μg/ml,线性范围上限分别5g/ml和20g/ml,谷氨酸和谷氨酰胺标准测定的相对标准偏差(rela-tive standard derivatizations,RSDs)分别为2.2%和3.9%,样品测定的RSD分别为3.7%和4.7%,平均加标回收率分别在94.2%～106.0%和80.6%～116.0%之间。[结论]所建立的HPLC法灵敏、快速,2,4-二硝基氯苯的衍生产物稳定,适用于血清样品中谷氨酸和谷氨酰胺的测定。[Objective] To establish a method for determination of glutamic acid （Glu） and glutamine （Gin） in serum by reversed-phase High Performance Liquid Chromatography （HPLC） with 2,4-dinitrechlorobenzene （DNCB） derivatization. [Methods ] After proteins in serum were precipitated with trichloroacetic acid （TCA）, amino acids in serum was derivatized with 2,4-dinitrochlorobenzene （DNCB）, and contents of Glu and Gin were analyzed by HPLC. [Results] The detection limits of this method for Gln and Gln were 0.005ug/ml and 0.054ug/ml, and whose linear upper limits were 5mg/ml and 20mg/ml, respectively. The relative standard derivations （RSDs） of standards for Glu and Gin were 2.2% and 3.9%, respectively, and those of samples were 3.7% and 4.7%, respectively. The average recoveries of Glu and Gin were at the range of 94.2%-106.0% and 80.6%- 116.0%, respectively. [Conclusion] The HPLC with 2, 4-dinitrecldombenzene with sensitive and rapid adapts for determination of Glu and Gin in serum.

关 键 词：谷氨酸和谷氨酰胺 高效液相色谱法 2 4-二硝基氯苯 血清 

分 类 号：R286.0[医药卫生—中药学] R122.11[医药卫生—中医学]