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- Title
Synthesis and characterization of (R)-phenylephrine bonded non-porous polystyrene-divinylbenzene for fast separation of proteins by mixed-mode liquid chromatography.
- Authors
Kitprapiumpon, Nirawan; Kulkanjanatorn, Pisamai; Wongyai, Surapote
- Abstract
A mixed-mode liquid chromatography PSDVB-PE solid support based on (R)-phenylephrine bonded non-porous polystyrene-divinylbenzene (PSDVB) was prepared and characterized for the fast separation of proteins. The support consisting of both anion-exchange and hydrophobic moieties was synthesized by bonding of the (R)-phenylephrine to the non-porous PSDVB particles via hydroboration, bromination and nucleophilic substitution reactions. The support with a surface coverage of 420 µmol/g was characterized with regard to its physical and chromatographic properties. The reverse phase and anion-exchange behaviors of this new stationary phase were investigated by injection of acidic, neutral and basic molecules at a wide pH range (pH 2 to 11.0) of eluents. The retention behaviors of the tested compounds (e.g. ethylparaben, benzoic acid, benzene and aniline) were studied by modifications of pH values and amounts of organic modifiers in the mobile phase in an isocratic elution. Fast separation of six proteins (e.g. cytochrome c, immunoglobulin G, ribonuclease A, human serum albumin, ovalbumin, and conalbumin) within 3 min showed many advantages over non-modified PSDVB support. Applications for separation of proteins in papaya latex, egg white, and human serum were demonstrated. Compare to the reverse phase column, the mixed-mechanism column provided a flexible and versatile method for fast separation of proteins.
- Subjects
PROTEIN fractionation; LIQUID chromatography; RIBONUCLEASE A; SUBSTITUTION reactions; NUCLEOPHILIC substitution reactions; IMMUNOGLOBULIN G; OVALBUMINS; CYTOCHROME c
- Publication
Pharmaceutical Sciences Asia, 2020, Vol 47, Issue 4, p366
- ISSN
2586-8195
- Publication type
Article
- DOI
10.29090/psa.2020.04.019.0133