A 3 μm undecylenic acid-functionalized stationary phase (UAS) was prepared for the separation of nucleosides and nucleobases using per aqueous liquid chromatography (PALC) and hydrophilic interaction liquid chromatography (HILIC). The retention behaviors of nucleosides and nucleobases in PALC and HILIC modes were explored by adjusting parameters such as water content, buffer concentration, pH of the mobile phase and column temperature. The experimental data and separation chromatogram demonstrated that PALC could provide retention comparable to that of HILIC for nucleosides and nucleobases. Comparative studies using diluted adenosine solutions evaluated theoretical plates and peak shape for the same retention factors (between 0.25 and 5.0) in PALC and HILIC. There was no buffer component in the mobile phases used to operate the comparisons. HILIC mode is more efficient for adenosine than PALC mode at low retention factors. It's the exact opposite phenomenon for high retention factors. It is proposed that the mass transfer of adenosine between the UAS, the water-rich layer and the ACN-rich mobile phase in HILIC is relatively slow. Given the significant use of toxic ACN in HILIC, PALC emerges as a safer and more effective alternative for separating nucleosides and nucleobases.

中文翻译：

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核苷和核碱基在 3 μm 十一碳烯酸功能化硅胶柱上在水相色谱和亲水相互作用液相色谱分离模式下的保留行为

制备了 3 μm 十一碳烯酸官能化固定相 (UAS)，用于使用全水性液相色谱 (PALC) 和亲水相互作用液相色谱 (HILIC) 分离核苷和核碱基。通过调整水含量、缓冲液浓度、流动相 pH 值和柱温等参数，探讨了 PALC 和 HILIC 模式下核苷和核碱基的保留行为。实验数据和分离色谱图表明，PALC 可以提供与 HILIC 相当的核苷和核碱基保留。使用稀释腺苷溶液的比较研究评估了 PALC 和 HILIC 中相同保留因子（0.25 至 5.0 之间）的理论板数和峰形。用于进行比较的流动相中没有缓冲剂成分。在低保留因子下，HILIC 模式比 PALC 模式对腺苷更有效。对于高保留因子来说，这是完全相反的现象。提出在 HILIC 中，UAS、富水层和富乙腈流动相之间腺苷的传质相对较慢。鉴于 HILIC 中大量使用有毒乙腈，PALC 成为分离核苷和核碱基的更安全、更有效的替代方案。