Imidazolium-embedded cysteine-functionalized stationary phases: preparation and its chromatographic applications

doi:10.3969/j.issn.1000-1565.2020.01.004

Abstract

Abstract: A novel imidazolium-embedded cysteine-functionalized stationary phase SIL-AVI-Cys was prepared by “click reaction”, which was further characterized by thermogravimetric analysis, FT-IR as well as elemental analysis. The stationary phase exhibits HILIC mode in the separation of five hydrophilic compounds, such as cytosine, cytidine, inosine, uracil and uridine, which also exhibits HILIC/RPLC mixed mode when separating anilines and phenolic substances. The separation mechanism of the stationary phase was further explored by investigating the effects of mobile phase pH and salt concentration on the retention of analytes. The change in the pH of the mobile phase can affect the interaction between analytes and the stationary phase by varying the dissociation of the analytes. The separation characteristics of the SIL-AVI-Cys column were also investigated by amides and nucleobases/nucleosides. The SIL-AVI-Cys column- DOI:10.3969/j.issn.1000-1565.2020.01.004咪唑嵌合半胱氨酸修饰固定相制备及色谱应用王胜玉¹,梁鹏¹,段立广²,乔晓强¹(1. 河北大学药学院,河北保定 071002;2. 河北医科大学第一医院药学部,河北石家庄 050000)摘要:利用“点击反应”合成了咪唑嵌合半胱氨酸硅胶固定相SIL-AVI-Cys,并通过热重、红外光谱以及元素分析对材料进行了表征.填充SIL-AVI-Cys色谱柱在分离胞核嘧啶、胞苷、肌苷、尿嘧啶和尿苷等5种亲水性化合物时表现为亲水作用模式,而在分离苯胺和苯酚类物质时则可表现出亲水/反相混合模式特征.通过考察流动相pH值、盐浓度对分析物保留的影响,进一步探讨了该固定相的分离机理.流动相pH值的改变可通过改变分析物的离子化影响其与固定相之间的作用力及其保留行为.流动相中盐浓度的增加,可减弱酸性分析物的保留,而对于碱性和中性分析物则无显著影响.此外,SIL-AVI-Cys色谱柱的分离性能采用酰胺及核酸碱基/核苷类物质进行了表征,并将其进一步应用于奶粉中三聚氰胺的表征和检测.关键词:混合模式作用色谱;氨基酸离子液体;固定相;奶粉;三聚氰胺中图分类号:O657.7 文献标志码:A 文章编号:1000-1565(2020)01-0018-09Imidazolium-embedded cysteine-functionalized stationary phases: preparation and its chromatographic applicationsWANG Shengyu¹, LIANG Peng¹, DUAN Liguang², QIAO Xiaoqiang¹(1. College of Pharmacy, Hebei University, Baoding 071002, China; 2. Department of Pharmacy, The First Hospital of Hebei Medical University, Shijiazhuang 050000, China)Abstract: A novel imidazolium-embedded cysteine-functionalized stationary phase SIL-AVI-Cys was prepared by “click reaction”, which was further characterized by thermogravimetric analysis, FT-IR as well as elemental analysis. The stationary phase exhibits HILIC mode in the separation of five hydrophilic compounds, such as cytosine, cytidine, inosine, uracil and uridine, which also exhibits HILIC/RPLC mixed mode when separating anilines and phenolic substances. The separation mechanism of the stationary phase was further explored by investigating the effects of mobile phase pH and salt concentration on the retention of analytes. The change in the pH of the mobile phase can affect the interaction between analytes and the stationary phase by varying the dissociation of the analytes. The separation characteristics of the SIL-AVI-Cys column were also investigated by amides and nucleobases/nucleosides. The SIL-AVI-Cys column- 收稿日期:2019-04-23 基金项目:国家自然科学基金资助项目(21675039) 第一作者:王胜玉(1990—),男,河北霸州人,河北大学硕士研究生,主要从事色谱分析研究. E-mail: 983942430@qq.com 通信作者:段立广(1984—),男,河北邢台人,河北医科大学第一医院主管药师,主要从事药物新剂型相关研究. E-mail: dlg123456@163.com乔晓强(1982—),男,河北宁晋人,河北大学教授,博士,主要从事色谱分离分析新材料研究. E-mail: hbuqiao@163.com第1期王胜玉等:咪唑嵌合半胱氨酸修饰固定相制备及色谱应用was further used for characterization and detection of melamine in baby milk powder.

Key words: mixed-mode chromatography, amino acid ionic liquid, stationary phase, baby milk powder, melamine

CLC Number:

O657.7

WANG Shengyu, LIANG Peng, DUAN Liguang, QIAO Xiaoqiang. Imidazolium-embedded cysteine-functionalized stationary phases: preparation and its chromatographic applications[J]. Journal of Hebei University (Natural Science Edition), 2020, 40(1): 18-26.

References

[1] REN X J, HU C X, GAO D, et al. Preparation of a poly(ethyleneimine)embedded phenyl stationary phase for mixed-mode liquid chromatography[J]. Analytica Chimica Acta, 2018, 1042: 165-173. DOI:org/10.1016/j.aca.2018.09.049.
[2] QU Q S, SI Y, XUAN H, et al. A nanocrystalline metal organic framework confined in the fibrous pores of core-shell silica particles for improved HPLC separation[J]. Microchimica Acta, 2017, 184(10): 4099-4106. DOI:10.1007/s00604-017-2439-1
[3] ZHANG L, DAI Q, QIAO X Q, et al. Mixed-mode chromatographic stationary phases: Recent advancements and its applications for high-performance liquid chromatography[J]. TrAC Trends in Analytical Chemistry, 2016, 82: 143-163. DOI:10.1016/j.trac.2016.05.011
[4] ZHANG B Y, LEI X Y, DENG L J, et al. Ultrafast preparation of a polyhedral oligomeric silsesquioxane-based ionic liquid hybrid monolith via photoinitiated polymerization, and its application to capillary electrochromatography of aromatic compounds[J]. Microchimica Acta,2018, 185(7): 318. DOI:10.1007/s00604-018-2847-x.
[5] QIAO X Q, ZHANG L, ZHANG N, et al. Imidazolium embedded C8 based stationary phase for simultaneous reversed-phase/hydrophilic interaction mixed-mode chromatography[J]. Journal of Chromatography A, 2015, 1400: 107-116. DOI:10.1016/j.chroma.2015.04.060.
[6] ZHAO W J, JIANG X M, NI S J, et al. Layer-by-layer self-assembly of polyelectrolyte multilayers on silica spheres as reversed-phase/hydrophilic interaction mixed-mode stationary phases for high performance liquid chromatography[J]. Journal of Chromatography A, 2017, 1499: 111-117. DOI:10.1016/j.chroma.2017.03.083.
[7] WANG Q, YE M, XU L, et al. A reversed-phase/hydrophilic interaction mixed-mode C18-Diol stationary phase for multiple applications[J]. Analytica Chimica Acta, 2015, 888: 182-190. DOI:10.1016/j.aca.2015.06.058.
[8] MCCALLUM J L, YANG R, YOUNG J C, et al. Improved high performance liquid chromatographic separation of anthocyanin compounds from grapes using a novel mixed-mode ion-exchange reversed-phase column[J]. Journal of Chromatography A, 2007, 1148(1): 38-45. DOI:10.1016/j.chroma.2007.02.088.
[9] BO C M, WANG C Z, WEI Y M. Preparation and evaluation of diblock copolymer-grafted silica by sequential surface initiated-atom transfer radical polymerization for reverse-phase/ion-exchange mixed-mode chromatography[J]. Journal of Separation Science, 2017, 40(24): 4700-4708. DOI:10.1002/jssc.201700719.
[10] WANG J B, LUE H X, LIN X C, et al. Monolithic column with double mixed-modes of hydrophilic interaction/cation-exchange and reverse-phase/cation-exchange stationary phase for pressurized capillary electrochromatography[J]. Electrophoresis, 2008, 29(4): 928-935. DOI:10.1002/elps.200700600.
[11] TAKAFUJI M, SHAHRUZZAMAN M, SASAHARA K, et al. Preparation and characterization of a novel hydrophilic interaction/ion exchange mixed-mode chromatographic stationary phase with pyridinium-based zwitterionic polymer-grafted porous silica[J]. Journal of Separation Science, 2018, 41(21): 3957-3965. DOI:10.1002/jssc.201800578.
[12] BO C M, WANG X M, WANG C Z, et al. Preparation of hydrophilic interaction/ion-exchange mixed-mode chromatographic stationary phase with adjustable selectivity by controlling different ratios of the co-monomers[J]. Journal of Chromatography A, 2017, 1487: 201-210. DOI:10.1016/j.chroma.2017.01.061.
[13] ZHOU D, ZENG J, FU Q F, et al. Preparation and evaluation of a reversed-phase/hydrophilic interaction/ion-exchange mixed-mode chromatographic stationary phase functionalized with dopamine-based dendrimers[J]. Journal of Chromatography A, 2018, 1571: 165-175. DOI:10.1016/j.chroma.2018.08.018.
[14] KAZARIAN A A, NESTERENKO P N, SOISUNGNOEN P, et al. Comprehensive analysis of pharmaceutical products using simultaneous mixed-mode(ion-exchange/reversed-phase)and hydrophilic interaction liquid chromatography[J]. Journal of Separation Science, 2014, 37(16):2138-2144. DOI:10.1002/jssc.201400411.
[15] LIU S, XU H, YU J, et al. Novel imidazolium-embedded N,N-dimethylaminopropyl-functionalized silica-based stationary phase for hydrophilic interaction/reversed-phase mixed-mode chromatography[J]. Analytical and Bioanalytical Chemistry, 2015, 407(30): 8989-8997. DOI:10.1007/s00216-015-9064-9.
[16] QIAO X, ZHANG L, ZHANG N, et al. Imidazolium embedded C8 based stationary phase for simultaneous reversed-phase/hydrophilic interaction mixed-mode chromatography[J]. Journal of Chromatography A, 2015, 1400: 107-116. DOI:10.1016/j.chroma.2015.04.060.
[17] LI Y, FENG Y, CHEN T, et al. Imidazoline type stationary phase for hydrophilic interaction chromatography and reversed-phase liquid chromatography[J]. Journal of Chromatography A, 2011, 1218(35): 5987-5994. DOI:10.1016/j.chroma.2011.04.023.
[18] QIAO L, SHI X, LU X, et al. Preparation and evaluation of surface-bonded tricationic ionic liquid silica as stationary phases for high-performance liquid chromatography[J]. Journal of Chromatography A, 2015, 1396: 62-71. DOI:10.1016/j.chroma.2015.03.081.
[19] OHYAMA K, TAKASAGO S, KISHIKAWA N, et al. Poly(L-lactic acid)-modified silica stationary phase for reversed-phase and hydrophilic interaction liquid chromatography[J]. Journal of Separation Science, 2015, 38(5): 720-723. DOI:10.1002/jssc.201401243.
[20] OHYAMA K, INOUE Y, KISHIKAWA N, et al. Preparation and characterization of surfactin-modified silica stationary phase for reversed-phase and hydrophilic interaction liquid chromatography[J]. Journal of Chromatography A, 2014, 1371: 257-260. DOI:10.1016/j.chroma.2014.10.073.
[21] IVERSON C D, LUCY C A. Aniline-modified porous graphitic carbon for hydrophilic interaction and attenuated reverse phase liquid chromatography[J]. Journal of Chromatography A, 2014, 1373: 17-24. DOI:10.1016/j.chroma.2014.11.003.
[22] QIU H, ZHANG M, GU T, et al. A sulfonic-azobenzene-grafted silica amphiphilic material: a versatile stationary phase for mixed-mode chromatography[J]. Chemistry-A European Journal, 2013, 19(52): 18004-18010. DOI:10.1002/chem.201302746.
[23] LIANG T, FU Q, SHEN A, et al. Preparation and chromatographic evaluation of a newly designed steviol glycoside modified-silica stationary phase in hydrophilic interaction liquid chromatography and reversed phase liquid chromatography[J]. Journal of Chromatography A, 2015, 1388: 110-118. DOI:10.1016/j.chroma.2015.02.019.
[24] WANG Q, YE M, XU L, et al. A reversed-phase/hydrophilic interaction mixed-mode C18-Diol stationary phase for multiple applications[J]. Analytica Chimica Acta, 2015, 888, 182-190. DOI:10.1016/j.aca.2015.06.058.
[25] WANG Q, LONG Y, YAO L, et al. Preparation, characterization and application of a reversed phase liquid chromatography/hydrophilic interaction chromatography mixed-mode C18-DTT stationary phase[J]. Talanta, 2016, 146: 442-451. DOI:10.1016/j.talanta.2015.09.009.
[26] QIU H, JIANG Q, WEI Z, et al. Preparation and evaluation of a silica-based 1-alkyl-3-(propyl-3-sulfonate)imidazolium zwitterionic stationary phase for high-performance liquid chromatography[J]. Journal of Chromatography A, 2007, 1163(1-2): 63-69. DOI:10.1016/j.chroma.2007.06.001.
[27] ZHANG L, DAI Q, QIAO X, et al. Mixed-mode chromatographic stationary phases: Recent advancements and its applications for high-performance liquid chromatography[J]. TrAC Trends in Analytical Chemistry, 2016, 82: 143-163. DOI:10.1016/j.trac.2016.05.011.
[28] WANG H, ZHANG L, MA T, et al. Imidazolium-embedded iodoacetamide-functionalized silica-based stationary phase for hydrophilic interaction/reversed-phase mixed-mode chromatography[J]. Journal of Separation Science, 2016, 39(18): 3498-3504. DOI:10.1002/jssc.201600448.
[29] QIU H, WANG L, LIU X, et al. Preparation and characterization of silica confined ionic liquids as chromatographic stationary phases through surface radical chain-transfer reaction[J]. Analyst, 2009, 134(3): 460-465. DOI:10.1039/B809125K.
[30] QIAO L, WANG S, LI H, et al. A novel surface-confined glucaminium-based ionic liquid stationary phase for hydrophilic interaction/anion-exchange mixed-mode chromatography[J]. Journal of Chromatography A, 2014, 1360: 240-247. DOI:10.1016/j.chroma.2014.07.096.