核酸碱基和核苷的高效液相色谱——表面增强拉曼光谱在线联机分析研究

Analysis of Nucleobases and Nucleosides by Reverse Phase-High Performance Liquid Chromatography Using On-Line Detection by Surface-Enhanced Raman Spectroscopy

本文将反相高效液相色谱(RP——HPLC)与表面增强拉曼光谱(SERS)联机,对三个核嘧啶、四个核嘌呤和两个核嘧啶及其核苷混合物进行在线联机分析研究。在流动条件下,对影响 SERS 测定的因素,如激光照射、温度、pH 值、流速、记忆效应,以及联机系统的接口装置等进行了讨论。文中给出了联机测得的三维和二维 SERS 色谱图。

The identification of a mixture of three nucleic pyrimidine bases (cytosine, uracil and thymine),a mixture of four purine bases (guanine adenine,xanthine and hy- poxanthine) and a mixture of two pyrimidine bases and two pyrimidine nucleosides (cy- tosine,uracil,cytidine,uridine) with reverse phase-high performance liquid chromatog- raphy (RP-HPLC) in combination with on -line surface-enhanced Raman spectroscopy (SERS) detection is demanstratecl.The factors which effect on SERS detection under flowing condition,such as the laser irradiation,temperature,pH,flow rate,memory effect,as well as the construcion of the interface between RP-HPLC system and Raman spectrometer was investigated.The quantity used for detecction was l nmol for adenine and guanine,2.5 nmol for cytosine and uracil,5 nmol for xanthine,cytidine and uri- dine,10 nmol for hypoxanthine and thymine.The three dimensional (SERS intensity, Raman shift and chromatographic retention time ) and two dimensional (SERS intensi- ty,and chromatographic retention time ) SERS chromatograms are presented.