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本工作将DANS反应和微量薄膜层析技术与荧光测量加以修改后联用,并注意了组织样品的去蛋白和DANS-牛磺酸洗脱剂的改进,获得了满意的定量测定氨基酸的结果。GABA、牛磺酸的浓度(GABA:0—60×10~(-12)克分子;牛磺酸:0—300×10~(-12)克分子)与其相应的DANS衍生物的荧光强度成线性关系。回收率分别为95—113％和82—94％。灵敏度比原有荧光方法提高了近100倍(10~(-12)克分子数量级)。用此法测得小鼠尾壳核中的GABA和牛磺酸分别为1.26±0.06和15.5±1.0微克分子/克湿重,组织样品只需0.5毫克或更少些。这为测定微量神经组织中的自由氨基酸提供了一种简便的超微量方法.
In this work, DANS reaction and micro-thin-layer chromatography techniques were used in combination with fluorescence measurements, and the deproteinization and DANS-taurine eluent improvements of the tissue samples were noted. Satisfactory quantitative results of the amino acids were obtained. The fluorescence intensity of GABA and taurine (GABA: 0-60 × 10-12 molecules; taurine: 0-300 × 10-12 molecules) and their corresponding DANS derivatives Linear relationship. The recoveries were 95-113% and 82-94%, respectively. Sensitivity than the original fluorescence method increased by nearly 100 times (10 ~ (-12) mole fraction). Using this method, GABA and taurine in the caudal putamen of mice were found to be 1.26 ± 0.06 and 15.5 ± 1.0 μg / g wet weight, respectively, and only 0.5 mg or less tissue samples were required. This provides a convenient ultra-trace method for the determination of free amino acids in trace nerve tissue.