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为了建立并改良高效液相色谱法测定大鼠基底核氨基酸类神经递质含量,我们改良使用0.1M磷酸二氢钾[KH2PO4,含0.8%四氢呋喃(THF),pH=5.8,流动相A]与甲醇∶乙腈=3∶1(流动相B)为流动相,梯度洗脱,邻苯二甲醛柱前衍生高效液相色谱荧光检测器检测大鼠基底核4种氨基酸类神经递质:谷氨酸(Glu)、谷氨酰胺(Gln)、甘氨酸(Gly)和γ-氨基丁酸(GABA)的含量。结果显示,4种氨基酸在18min内洗脱完全,分离效果良好;其相关系数均为0.999;4种氨基酸1天内精密度范围为1.85%～7.95%,绝对回收率均>86.30%。提示,改良的高效液相色谱荧光法灵敏度高、重复性好,能有效分离检测大鼠基底核氨基酸类神经递质含量。
In order to establish and improve the HPLC method for the determination of amino acid neurotransmitters in rat basal nuclei, we modified and purified 0.1M potassium dihydrogen phosphate [KH2PO4 (containing 0.8% tetrahydrofuran (THF), pH = 5.8, mobile phase A] and Methanol: acetonitrile = 3: 1 (mobile phase B) was used as the mobile phase and the gradient elution was performed. The four amino acid neurotransmitters, glutamic acid (Glu), glutamine (Gln), glycine (Gly) and γ-aminobutyric acid (GABA) content. The results showed that the four kinds of amino acids eluted completely within 18min, the separation effect was good. The correlation coefficients were 0.999. The precision of four amino acids within 1 day ranged from 1.85% to 7.95% and the absolute recoveries were both> 86.30%. It is suggested that the improved high performance liquid chromatography with high sensitivity and repeatability can effectively separate and detect the content of amino acid neurotransmitters in rat basal nucleus.