Physically adsorbed coatings based on chitosan for electrophoretic separation of biologically active substances

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Abstract

Coatings of the inner walls of a quartz capillary based on cationic high-molecular chitosan with a deacetylation degree of 95% were formed. The dependence of the electroosmotic flow rate on the pH of the background electrolyte was studied, and the stability of the coating under the influence of various solvents was assessed. The results were compared with another cationic coating based on poly(diallyldimethylammonium chloride) (PDADMAC). It was shown that when separating amino acids, catecholamines, and organic acids, the formed coatings based on chitosan are slightly inferior in efficiency to coatings made of PDADMAC, but provide a higher resolution of the studied biologically active analytes. It was found that chitosan on the inner walls of a quartz capillary promotes an increase in enantioselectivity in the separation of β-blocker enantiomers (carvedilol, propranolol, sotalol) in the presence of (2-hydroxypropyl)-β-cyclodextrin in the background electrolyte, as well as non-steroidal anti-inflammatory drugs (ketoprofen and ketorolac) using vancomycin as the second chiral selector.

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E. A. Kolobova

Institute of Chemistry of the Saint Petersburg State University

Author for correspondence.
Email: ekatderyabina@mail.ru
Russian Federation, 26, Universitetsky Ave., Peterhof, Saint Petersburg, 198504

E. R. Ziangirova

Institute of Chemistry of the Saint Petersburg State University

Email: ekatderyabina@mail.ru
Russian Federation, 26, Universitetsky Ave., Peterhof, Saint Petersburg, 198504

E. V. Solovyova

Institute of Chemistry of the Saint Petersburg State University

Email: ekatderyabina@mail.ru
Russian Federation, 26, Universitetsky Ave., Peterhof, Saint Petersburg, 198504

L. A. Kartsova

Institute of Chemistry of the Saint Petersburg State University

Email: ekatderyabina@mail.ru
Russian Federation, 26, Universitetsky Ave., Peterhof, Saint Petersburg, 198504

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Supplementary files

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2. Scheme 1. Structure of chitin/chitosan. n is the number of repeating units of glucosamine, m is the number of repeating units of acetylglucosamine, n + m is the degree of polymerisation, n / (m + n) is the degree of deacetylation. When n exceeds 50% of the total number of links, the polymer is called chitosan [17].

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3. Fig. 1. Dependences of electrophoretic mobility of EOP (µEOP) on the pH of the background electrolyte: 1 - uncoated capillary; 2 - capillary modified with PDADMAH [39]; 3 - chitosan-coated capillary.

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4. Fig. 2. SEM images of a cross section of a capillary coated with (a), (b) chitosan-based and (c) PDADMAH obtained at different magnifications with a scale bar: (a) 2 μm; (b), (c) 200 nm. Instrument: Zeiss Merlin, secondary electron image.

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5. Fig. 3. Electrophoregrams of a model mixture of organic acids: (a) capillary modified with PDADMAH; (b) capillary modified with chitosan. Background electrolyte: 10 mM benzoic acid, 1 mM EDTA, 10 mM DEA. Model mixture of carboxylic acids 25 µg/ml: 1 - oxalic acid, 2 - formic acid, 3 - tartaric acid, 4 - citric acid, 5 - malic acid, 6 - lactic acid, 7 - succinic acid, 8 - acetic acid, 9 - propionic acid, 10 - butyric acid.

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6. Fig. 4. Electrophoregrams of a model mixture of amino acids and catecholamines (50 µg/ml): (a) unmodified capillary; (b) capillary modified with chitosan; (c) capillary modified with PDADMAH. Background electrolyte: 10 mM phosphate buffer solution with pH 2.0. Analytes: 1 - DOPA, 2 - Tyr, 3 - Phe, 4 - Trp, 5 - E, 6 - NMN, 7 - NE, 8 - 3-MT, 9 - DA.

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7. Fig. 5. Electrophoregrams of the model mixture of β-blockers on different capillaries: (a)-(c) capillary modified with chitosan; (d), (e) unmodified capillary; (f) capillary coated with PDADMAH. Background electrolyte: (a), (d) 25 mM phosphate buffer solution (PBS) with pH 2.0; (b) 25 mM PBS with pH 2.0, 0.01 wt% chitosan; (c), (d), (e), (f) 25 mM PBS with pH 2.0, 5 mM GP-β-CD. Analyses: 1 - sotalol, 2 - propranolol, 3 - carvedilol.

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8. Fig. 6. Electrophoregrams of a model mixture of non-steroidal anti-inflammatory drugs (25 µg/ml). Droplet-105M; capillary modified with chitosan. Background electrolyte: (a) 25 mM phosphate buffer solution (PBS) with pH 6.4; (b) 25 mM PBS with pH 6.4, 0.01 wt% chitosan; (c) 25 mM PBS with pH 6.4, 0.5 mM GP-β-CD; (d) 25 mM PBS with pH 6. 4, 0.5 mM β-CD; (e) 25 mM FBI with pH 4.2, 2.5 mM vancomycin, 10 vol % methanol; (f) unmodified capillary, 25 mM FBI with pH 4.2, 2.5 mM vancomycin, 10 vol % methanol. Analyses: 1 - ketorolac, 2 - ketoprofen, 3 - ibuprofen.

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9. Fig. 7. Dependences of resolution (Rs) of enantiomers (1) of ketorolac and (2) of ketoprofen on the composition of the background electrolyte. Conditions: capillary modified with chitosan; background electrolyte: (a) 10-25 mM FBR with pH 4.2, 1 mM vancomycin; (b) 25 mM FBR with pH 4.2, 1 mM vancomycin, 0-20 vol. % methanol; (c) 25 mM FBR with pH 4.2, 0-5 mM vancomycin, 10 vol. % methanol. Analyses: 1 - ketorolac, 2 - ketoprofen.

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