Rapid and reliable method for the analysis of nucleotide pools by reversed-phase high-performance liquid chromatography.
Rapid and reliable method for the analysis of nucleotide pools by reversed-phase high-performance liquid chromatography.
复制标题
通过反相高效液相色谱分析核苷酸池的快速可靠的方法。
DOI：
10.1016/s0021-9673(00)85954-3
复制
发表时间：
1982
期刊：
影响因子：
--
通讯作者：
Taylor,MW
中科院分区：
文献类型：
--
作者：
Martinez-Valdez,H;Kothari,RM;Hershey,HV;Taylor,MW
本文建立了一种快速、可靠的同时分离核糖、脱氧核糖和环核苷酸的方法。该方法采用C18μBondapak柱，磷酸铵缓冲液（0.2M，pH5.1）等度洗脱。已通过在高碘酸盐氧化前后进行分离来确认脱氧核糖核苷酸的分离度。洗脱的一般顺序是核糖核苷酸、脱氧核糖核苷酸和环核苷酸。虽然高碘酸氧化通过消除核糖核苷酸提高了脱氧核糖核苷酸分离的清晰度，但在洗脱液中掺入甲醇缩短了环核苷酸的保留时间。应用这种方法对一个复杂的生物系统的报告。
A rapid and reliable protocol for the simultaneous separation of ribo-, deoxyribo- and cyclic nucleotides has been developed using high-performance liquid chromatography on a C18μBondapak column and isocratic elution with ammonium phosphate buffer (0.2M, pH 5.1). Resolution of deoxyribonucleotides has been confirmed by performing resolution before and after periodate oxidation. The general order of elution is ribonucleotides, deoxyribonucleotides and cyclic nucleotides. While periodate oxidation improved the clarity of separation of deoxyribonucleotides by eliminating ribonucleotides, incorporation of methanol in the eluent shortened the retention time of the cyclic nucleotides. The application of this method to a complex biological system is reported.