人胃粘膜中γ-氨基丁酸和谷氨酸含量的高效液相色谱法测定

Determination of γ-Aminobutyric Acid and Glutamate in Human Gastric Mucosa by High Performance Liquid Chromatography

建立了胃粘膜组织中γ 氨基丁酸和谷氨酸含量的高效液相色谱测定法。用异硫氰酸苯酯(PITC)作衍生剂,色谱柱为Pico·Tag 氨基酸专用柱,梯度洗脱,紫外检测波长254nm。γ 氨基丁酸和谷氨酸分别在0 125～6.25μmol/L,0 025～2 5mmol/L时线性关系良好;γ 氨基丁酸和谷氨酸的平均加样回收率分别为95 4%和93 5%,批内误差分别为3 56%和1 12%,批间误差分别为7 47%和5 98%。该方法稳定、灵敏。用该方法测定了30例胃粘膜标本,结果胃癌组织的γ 氨基丁酸和谷氨酸含量显著高于正常组织。

A method for analysis of γ-aminobutyric acid (GABA) and glutamate (Glu) in human gastric mucosa by high performance liquid chromatography (HPLC) was developed. A gradient elution was used for the separation and quantification of GABA and Glu after pre-column derivatization with phenylisothiocyanate (PITC). The column was Pico·Tag~ for free amino acids. GABA and Glu were determined with UV detector at 254 nm. Good linearities were observed within the ranges from 0.125 to 6.25 (μmol/L) for GABA and from 0.025 to 2.5 (mmol/L) for Glu. The average recoveries were 95.4% for GABA and 93.5% for Glu. The intra- and inter-precision values were within 3.56% and 7.47% for GABA, and 1.12% and 5.98% for Glu, respectively. The method is sensitive, specific and accurate. It can be used in the determination of GABA and Glu in human gastric mucosa tissue. The concentrations of GABA and Glu in cancer tissues are significantly higher than those in normal tissue.