HPLC法测定大鼠脑组织游离天门冬氨酸、谷氨酸、肌肽及γ-氨基丁酸  

Determination of Free Aspartate,Glutamate,Carnosine and GABA in Rat Brain by HPLC

作　　者：宋杨[1] 于涛[1] 庞磊[1] 刘影[1] 韩璐[1] 杨菁[1] 

机构地区：[1]辽宁医学院生物化学教研室,辽宁锦州121000

出　　处：《辽宁医学院学报》2010年第4期314-316,共3页Journal of Liaoning Medical University (LNMU) Bimonthly

基　　金：辽宁省教育厅创新团队资助;编号:2008T115

摘　　要：目的建立镄4定大鼠脑组织中游离天门冬氨酸、谷氨酸、肌肽以及γ-氨基丁酸含量的高效液相色谱法。方法采用C18色谱柱，以乙腈和25mmol／L磷酸盐缓冲液（pH6．8）为流动相，用邻苯二甲醛为柱前衍生剂，流速为1mL／min，激发波长为338nln，发射波长为425nm，在柱温38℃下采用二元梯度洗脱，同时对大鼠脑组织中游离天门冬氨酸、谷氨酸、肌肤以及γ-氨基丁酸的含量进行了测定。结果天门冬氨酸、谷氨酸、肌肽以及γ-氨基丁酸在该色谱奈件下均可基线分离，在O．5—6μg／mL范围内呈现良好线性关系，天门冬氨酸、谷氨酸、肌肽以及γ-氨基丁酸的相关系数分别是0．9991，0．9998，0．9996，0．9995。回收率分别为99．80％，99．74％，99．63％，99．34％。结论该方法灵敏可靠，专属性强，重现性好。天门冬氨酸、谷氨酸、肌肽以及γ-氨基丁酸在大鼠脑组织中的含量分别为0．77±0．04、1．16±0．07、0．36±0．05、1．30±0．06mg／g。Objective To establish HPLC methods to determine free Aspartate, Glutamate, Camosine and GABA in brain. Methods The sample was derived from O-phthalaldehyde （OPA）, and then the derivative sample was separated on Kromail C18 column at 38℃. A linear gradient elution of A （0. 025 mol/L phosphate buffer of 3% Acetonitrile, pH 6. 8） and B （0. 025 moiL/ L phosphate buffer of 50% Acetonitrile） was used, the flow rate was 1 mL/min, and detected by fluorescence detector with excitation wavelength （kex） at 338 run and emission wavelength （λem） at 425 nm. Results Aspartate, Glutamate, Carnosine and GABA were separated to baseline. A good linearity was obtained over the range of 0. 5 to 6 μg/mL, and the correlation coefficients of Aspartate, Glutamate, Carnosine and GABA were O. 9991,0. 9998, 0. 9996 and 0. 9995, respectively, the recoveries were 99. 34% -99. 80%. Conclusions The result shows that this method is simple, efficient, specific, and reliable. The contents of free Aspartate, Glutamate, camosine and GABA in rat＇s brain were （0. 77±0. 04）, （ 1.16±0. 07）, （0. 36±0. 05）, （ 1.30±0. 06） mg/g.

关 键 词：天门冬氨酸 谷氨酸 Γ-氨基丁酸 肌肽 邻苯二甲醛 

分 类 号：Q593[生物学—生物化学]