色谱 ›› 2012, Vol. 30 ›› Issue (10): 1062-1067.DOI: 10.3724/SP.J.1123.2012.06028

• 研究论文 • 上一篇    下一篇

厚朴酚键合硅胶高效液相色谱固定相分离嘌呤、嘧啶、蝶呤及黄酮

陈红, 李来生*, 张杨, 周仁丹   

  1. 南昌大学分析测试中心, 江西 南昌 330047
  • 收稿日期:2012-06-18 修回日期:2012-08-13 出版日期:2012-10-28 发布日期:2012-10-17
  • 通讯作者: 李来生,博士,教授,主要研究方向为色谱分析与生化分析研究. Tel: (0791)88304414, E-mail: lilaishengcn@163.com.
  • 基金资助:

    江西省自然科学基金项目(2010GZH0089).

Separation of purines, pyrimidines, pterins and flavonoids on magnolol-bonded silica gel stationary phase by high performance liquid chromatography

CHEN Hong, LI Laisheng*, ZHANG Yang, ZHOU Rendan   

  1. Center of Analysis and Testing, Nanchang University, Nanchang 330047, China
  • Received:2012-06-18 Revised:2012-08-13 Online:2012-10-28 Published:2012-10-17

摘要: 将新制备的厚朴酚键合硅胶固定相(MSP)用于嘌呤、嘧啶、蝶呤及黄酮类化合物的液相色谱分离分析。选取了4种嘌呤、8种嘧啶、4种蝶呤及5种黄酮类药物作为极性化合物的代表,以商品反相碳十八烷基键合硅胶柱(ODS)作参照,研究了新固定相对碱性化合物的选择性和相关分离机理。实验发现,在简单流动相条件下,厚朴酚键合硅胶固定相对上述药物表现出较高的选择性及分离效果。尽管MSP没有进行封尾处理,但含氮类极性化合物(嘌呤、嘧啶、蝶呤)仍表现出基本对称的色谱峰形。多数药物在两柱上的洗脱顺序大致相同,说明疏水作用始终存在,这说明新固定相具有反相色谱性能。比较研究还发现,MSP在分离上述极性药物时能够提供除疏水性作用之外的其他作用位点。例如,在分离嘌呤、嘧啶及蝶呤时,氢键和偶极作用明显存在;同时MSP与溶质结构中的芳环(硫唑嘌呤、紫花牡荆素)之间有较强的π-π电子相互作用等,使得含氮类极性化合物和黄酮的保留一般比ODS强,分离度也有一定的改善。多种作用可以合理地解释MSP柱对极性溶质有更强的分离能力,厚朴酚键合硅胶固定相可在一定程度上弥补ODS单一疏水作用的不足,有利于分类碱性化合物。

关键词: 蝶呤, 反相高效液相色谱, 厚朴酚键合硅胶固定相, 黄酮, 嘧啶, 嘌呤, 色谱保留机理

Abstract: A new magnolol-bonded silica gel stationary phase (MSP) was used to separate the basic drugs including four purines, eight pyrimidines, four pterins and five flavonoids as polar representative samples by high performance liquid chromatography (HPLC). To clarify the separation mechanism, a commercial ODS column was also tested under the same chromatographic conditions. The high selectivities and fast baseline separations of the above drugs were achieved by using simple mobile phases on MSP. Although there is no end-caped treatment, the peak shapes of basic drugs containing nitrogen such as purines, pyrimidines and pterins were rather symmetrical on MSP, which indicated the the magnolol as ligand with multi-sites could shield the side effect of residual silanol groups on the surface of silica gel. Although somewhat different in the separation resolution, it was found that the elution orders of some drugs were generally similar on both MSP and ODS. The hydrophobic interaction should play a significant role in the separations of the above basic drugs, which was attributed to their reversed-phase property in the nature. However, MSP could provide the additional sites for many polar solutes, which was a rational explanation for the high selectivity of MSP. For example, in the separation of purines, pyrimidines and pterins on MSP, hydrogen-bonding and dipole-dipole interactions played leading roles besides hydrophobic interaction. Some solute molecules (such as mercaptopurine, vitexicarpin) and MSP can form the strong π-π stacking in the separation process. All enhanced the retention and improved the separation selectivity of MSP, which facilitated the separation of the basic drugs.

Key words: chromatographic retention mechanism, flavonoids, pterins, purines, pyrimidines, reversed-phase high performance liquid chromatography (RP-HPLC), magnolol-bonded silica gel stationary phase

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