含腺嘌呤化合物的高效液相色谱灵敏荧光分析法
Sensitive fluorimetry of adenine-containing compounds with high-performance liquid chromatography.
作者信息
Yoshioka M, Nishidate K, Iizuka H, Nakamura A, El-Merzabani M M, Tamura Z, Miyazaki T
出版信息
J Chromatogr. 1984 Jul 13;309(1):63-71. doi: 10.1016/0378-4347(84)80006-7.
A definitive method to determine adenine compounds simultaneously was established by introducing a new fluorescent reagent into high-performance liquid chromatography. Bromoacetaldehyde was the best reagent among the haloacetaldehydes examined. A quantitative reaction was obtained even for unstable ADP and ATP. A high resolution of adenine nucleotides was obtained using a column of Hitachi gel No. 3012-N. The method was applied to the measurement of cyclic AMP in urine, and ADP and ATP in brain and blood. Further, the sensitivity of the method was increased by a new fluorescence spectrophotometer constructed for micro-HPLC. Femtomole amounts of the adenine nucleotides were clearly separated.
通过将一种新型荧光试剂引入高效液相色谱法，建立了一种同时测定腺嘌呤化合物的确定性方法。在所研究的卤代乙醛中，溴乙醛是最佳试剂。即使对于不稳定的ADP和ATP也能实现定量反应。使用日立3012-N号凝胶柱可实现腺嘌呤核苷酸的高分辨率分离。该方法应用于尿液中环磷酸腺苷以及脑和血液中ADP和ATP的测定。此外，通过为微高效液相色谱构建的新型荧光分光光度计提高了该方法的灵敏度。飞摩尔量的腺嘌呤核苷酸能够清晰分离。
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