RP-HPLC法检测大鼠海马中4种氨基酸类神经递质含量的研究

Determination of 4 Amino Acids Neurotransmitters in Rat Hippocampi by RP-HPLC

建立了RP-HPLC测定大鼠海马中4种氨基酸类神经递质含量的方法。以2,4-二硝基氟苯（DNFB）为衍生化试剂进行柱前衍生,色谱条件为Diamonsil C18（4.6mm×200mm,5μm）色谱柱,乙腈和醋酸钠缓冲液为流动相,采用二元梯度洗脱,UV350nm检测。结果显示：在18min内,4种氨基酸分离完全,在0.01-100μg/mL范围内线性关系良好,天冬氨酸（Asp）、谷氨酸（Glu）、甘氨酸（Gly）、γ-氨基丁酸（GABA）相关系数分别为0.9985、0.9976、0.9943、0.9924;日内变异系数分别为2.4%、3.8%、3.2%和3.5%;标准加入平均回收率分别为94.80%、93.93%、92.69%和96.74%。

In this paper,a method for the determination of 4 amino acids neurotransmitters in rat hippocampi by RP-HPLC was established. The samples were precolumn derivated with DNFB. The chromatogram condition is Diamonsil C18 column （4.6mm×200mm, 5μm）. Acetonitrile/acetate buffer solution was used as mobile phase and the gradient elution was detected by UV at 350 nm. The results showed that 4 amino acids neurotransmitters were separated within 18 minutes. The linear relation was well in the range from 0.01 to 100 μg/mL,and the correlation coefficients of Asp,Glu,Gly and GABA were 0. 9985,0. 9976,0. 9943,and 0. 992 4,respectively. The coefficient of differentiations were 2. 4%, 3. 8%, 3. 2% and 3. 5%, and the average recoveries were 94.80% ,93.93% ,92.69% and 96.74% ,respectively. This method is speediness,accurate,simple, high sensible and well repetition and provides a reference for the determination of amino acids neurotransmitter and related researches.