Shi Qian, Chen Junhui, Li Xin, Cao Wei, Zheng Li, Zang Jiaye, Wang Xiaoru
Qingdao Key Lab of Analytical Technology and Standardization of Chinese Medicines, The First Institute of Oceanography, State Oceanic Administration, Qingdao 266061, China.
Se Pu. 2011 Jun;29(6):481-7. doi: 10.3724/sp.j.1123.2011.00481.
The most suitable background electrolytes (BGEs) for simultaneous separation of 15 nucleosides by different modes of capillary electrophoresis (CE) were obtained. Various modes of CE were performed including capillary zone electrophoresis (CZE), capillary electrophoresis-electrospray ionization-time of flight mass spectrometry (CE-ESI-TOF/MS) and micellar electrokinetic chromatography (MEKC). The electrolyte buffers using sodium tetraborate decahydrate, disodium hydrogen phosphate, sodium acetate, sodium bicarbonate, ammonium acetate or 1, 2-diamino-ethane (DEA) were tested, and the best of them were systematically optimized. In CZE mode, the nucleosides could not be separated completely with sodium tetraborate decahydrate or disodium hydrogen phosphate as BGEs, demonstrating the limited applicability of the two buffer systems for complex samples. However, with 300 mmol/L DEA (containing 2% acetone) as BGE, 15 nucleosides could be separated with good resolution and peak shape, which proved that the DEA buffer was most suitable in CZE. The best buffer system in MEKC mode was 25 mmol/L disodium hydrogen phosphate with 70 mmol/L sodium dodecyl sulfate (SDS), and it was successfully applied for the separation of the nucleosides in Chinese Anthopleura lanthogrammica Berkly. The optimum buffer system for CE-ESI-TOF/MS analysis was 20 mmol/L ammonium acetate (pH 10.0). In the positive ion mode, the MS signals of each compound were better than those in the literature using DEA as BGE. The results of this study demonstrated the applicability of different buffer systems for the simultaneous separation of 15 nucleosides, and were helpful for the development of CE method in complex sample separation.
获得了用于通过不同模式的毛细管电泳(CE)同时分离15种核苷的最合适背景电解质(BGEs)。进行了多种CE模式,包括毛细管区带电泳(CZE)、毛细管电泳-电喷雾电离-飞行时间质谱(CE-ESI-TOF/MS)和胶束电动色谱(MEKC)。测试了使用十水合四硼酸钠、磷酸氢二钠、醋酸钠、碳酸氢钠、醋酸铵或1,2-二氨基乙烷(DEA)的电解质缓冲液,并对其中最佳的缓冲液进行了系统优化。在CZE模式下,以十水合四硼酸钠或磷酸氢二钠作为BGE时,核苷无法完全分离,这表明这两种缓冲系统对复杂样品的适用性有限。然而,以300 mmol/L DEA(含2%丙酮)作为BGE时,15种核苷能够以良好的分辨率和峰形分离,这证明DEA缓冲液在CZE中最为合适。MEKC模式下最佳的缓冲系统是25 mmol/L磷酸氢二钠与70 mmol/L十二烷基硫酸钠(SDS),并且它成功应用于中国海葵(Anthopleura lanthogrammica Berkly)中核苷的分离。CE-ESI-TOF/MS分析的最佳缓冲系统是20 mmol/L醋酸铵(pH 10.0)。在正离子模式下,每种化合物的质谱信号都优于文献中使用DEA作为BGE时的信号。本研究结果证明了不同缓冲系统用于同时分离15种核苷的适用性,有助于复杂样品分离中CE方法的发展。