一种聚合型弱阳离子交换/亲水相互作用色谱固定相的制备及色谱性能

doi:10.3724/SP.J.1123.2019.08019

摘要/Abstract

摘要：

通过亲核取代反应将聚乙烯马来酸酐键合到氨基硅胶表面,然后将残余的马来酸酐水解,制备了一种弱阳离子交换/亲水相互作用高效液相色谱固定相(Sil-PolyCOOH),通过固体核磁、ζ-电势及元素分析对固定相进行了表征。选取核苷和核酸碱为模型化合物,通过考察流动相组成,离子强度和pH等因素对溶质保留的影响,探讨了固定相的分离性能和保留机理,结果表明,该固定相的保留机理同时涉及亲水分配相互作用和多重主客体作用力。该固定相还对糖类、敌草快与百草枯等化合物具有良好的分离性能。上述研究结果表明该固定相在极性化合物的分离上具有良好的应用前景。

关键词: 亲水相互作用色谱, 高效液相色谱, 弱阳离子交换, 聚乙烯马来酸酐, 固定相

Abstract:

a weak cationic exchange/hydrophilic interaction mixed-mode stationary phase (Sil-PolyCOOH) for high performance liquid chromatography was prepared by bonding polyethylene maleic anhydride to the surface of amino silica gel via a nucleophilic substitution reaction, followed by hydrolysis of the residual maleic anhydride. The new stationary phase material was characterized by solid-state¹³C nuclear magnetic resonance analysis and ζ-potential measurements. The chromatographic performance and retention mechanism of this stationary phase were evaluated using nucleosides and nucleic bases as probes. The effects of mobile phase composition, ion strength, and pH on the retention factor were investigated. The results show that the retention mechanism of the stationary phase involves both hydrophilic distribution interaction and multiple forces between the host and guests. The stationary phase also showed good separation performance for saccharides, diquat, and paraquat. These results show that the stationary phase has a good application prospect in the separation of polar compounds.

Key words: hydrophilic interaction chromatography (HILIC), high performance liquid chromatography (HPLC), weak cation exchange, polyethylene maleic anhydride, stationary phase

中图分类号:

O658

娄旭华, 左慧颖, 王媛, 赵文杰. 一种聚合型弱阳离子交换/亲水相互作用色谱固定相的制备及色谱性能[J]. 色谱, 2020, 38(4): 430-437.

LOU Xuhua, ZUO Huiying, WANG Yuan, ZHAO Wenjie. Preparation and chromatographic evaluation of a polymer-based stationary phase for weak anion-exchange/hydrophilic interaction[J]. Chinese Journal of Chromatography, 2020, 38(4): 430-437.

图/表 11

图1 聚合物基弱阳离子交换/亲水相互作用色谱固定相的制备过程

Fig. 1 Schematic illustration of the synthetic process of weak anion-exchange/hydrophilic interaction stationary phase DMF: N,N-dimethylformamide.

图2 固定相Sil-PolyCOOH的固体13C CP/MAS NMR谱图

Fig. 2 Solid-state 13C CP/MAS NMR spectra of Sil-PolyCOOH material 13C CP/MAS NMR: 13C cross polarization magic angle spinning nuclear magnetic resonance spectroscopy; δ: chemical shift.

图3 Sil-PolyCOOH材料在不同pH条件下的ζ-电势

Fig. 3 ζ-potential of Sil-PolyCOOH material at different pH levels

图4 核苷和核酸碱类化合物在Sil-polyCOOH柱上的分离

Fig. 4 Separation of nucleosides and nucleic bases compounds on Sil-polyCOOH column Mobile phase: CH3CN-ammonium acetate (85∶15, v/v, pH 3.26); detection wavelength: 265 nm. Peak Nos.: 1. thymidine; 2. uridine; 3. inosine; 4. adenine; 5. 2-NH2-adenine nucleoside; 6. cytidine; 7. cytosine.

图5 流动相中水含量对核苷和核酸碱保留的影响

Fig. 5 Influence of water content in the mobile phase on the retention of nucleosides and nucleic bases k: retention factor; CB: water content in the mobile phase.

表1 基于保留模型(1)、(2)和(3)的线性回归系数

Table 1 Linear regression coefficients based on retention models (1), (2), and (3)

Solute	Retention model (1)			Retention model (2)			Retention model (3)
Solute	lg k_w	-S	R²	lg k_B	-A_S/n_B	R²	a	b	c	R²
Thymidine	0.109	3.177	0.975	-1.411	0.913	0.839	2.575	0.894	-11.312	0.986
Uridine	0.276	3.389	0.980	-1.346	0.976	0.846	3.003	0.911	-11.873	0.991
Inosine	0.587	3.723	0.965	-1.204	1.082	0.853	3.114	0.679	-11.608	0.964
Adenine	0.539	2.767	0.970	-0.826	0.847	0.973	-1.237	-0.953	-2.109	0.988
2-NH₂-adenine	0.686	3.147	0.980	-0.861	0.957	0.967	-0.646	-0.857	-3.416	0.992
Cytidine	0.870	3.763	0.987	-0.977	1.140	0.965	-0.279	-0.878	-4.740	0.995
Cytosine	0.874	3.215	0.947	-0.718	0.992	0.968	-1.513	-1.357	-1.336	0.972

图6 (a)流动相中缓冲盐浓度和(b)pH对模型化合物保留的影响

Fig. 6 Influence of (a) buffer concentration in the mobile phase and (b) pH values on the retention of model compounds

图7 模型化合物在Sil-polyCOOH柱上的范特霍夫曲线

Fig. 7 Van’t Hoff plots for model compounds on Sil-polyCOOH column Mobile phase: CH3CN-ammonium acetate (85∶15, v/v, pH 3.26); detection wavelength: 265 nm. T: thermodynamic temperature.

表2 Sil-polyCOOH柱上7个模型化合物的热力学参数

Table 2 Thermodynamic parameters for seven model compounds on Sil-polyCOOH column

Analyte	ΔH/ (kJ/mol)	ΔS/ (J/(mol·K))	Correlation coefficient
Thymidine	-7.67	-35.25	0.9996
Uridine	-8.05	-32.91	0.9988
Inosine	-7.56	-31.23	0.9987
Adenine	-7.30	-27.82	0.9998
2-NH₂-adenine	-7.96	-27.89	0.9942
Cytidine	-7.74	-25.31	0.9983
Cytosine	-9.03	-28.14	0.9977

图8 糖类化合物在Sil-polyCOOH柱上的分离

Fig. 8 Separation of saccharides on Sil-polyCOOH column Mobile phase: CH3CN/H2O (90∶10, v/v); ELSD (evaporative light scattering detection): gas pressure, 2×105 Pa; tube temperature, 80 ℃; gain, 30.

图9 百草枯和敌草快在Sil-polyCOOH柱上的分离

Fig. 9 Separation of paraquat and diquat on Sil-polyCOOH column Mobile phase conditions: ACN/15 mmol/L CH3COONH4 (75∶25, v/v); detection wavelength: 257 nm.

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