A Method for the Quantitation of Charge by Size Exclusion Chromatography Demonstrated with Components of Ficoll 400

Carboxylated Ficolls: Preparation, Characterization, and Electrophoretic Behavior of Model Charged Nanospheres

“…[12][13][14] Conversion of surface hydroxyl groups to carboxyl groups offers the possibility of a series of negatively charged spheres of variable size and charge, which should be model solutes for the study of electrophoretic mobility. In this paper, we report on the preparation of carboxylated ficoll samples with sizes between those of dendrimers and latex particles.…”

“…Ficolls, prepared by cross-linking polysucrose with epichlorhydrin (Figure ), are highly branched polysaccharides, whose spherical shape and size is retained regardless of solution ionic strength. − Conversion of surface hydroxyl groups to carboxyl groups offers the possibility of a series of negatively charged spheres of variable size and charge, which should be model solutes for the study of electrophoretic mobility. In this paper, we report on the preparation of carboxylated ficoll samples with sizes between those of dendrimers and latex particles.…”

Hyaluronic Acid as Macromolecular Crowder in Equine Adipose-Derived Stem Cell Cultures

“…With respect to biophysical analysis, all molecules assessed had a negative charge; the high negative charge of carrageenan can be attributed to its sulphate content [ 66 ], and the Ficoll ® , although it is considered to be non-ionic, is a rather heterogeneous in molecular weight macromolecule and has been demonstrated to contain negatively charged residues on about 50% of its macromolecular components [ 67 ], and the carboxyl groups of the glucuronic acid residues are negatively charged at physiological pH and ionic strength, making hyaluronic acid a negatively charged polyanionic macromolecule [ 68 , 69 , 70 ]. For the HA 100 kDa and the HA 500 kDa, a concentration-dependent zeta potential decrease and a hydrodynamic radius increase was observed as a function of increased concentration.…”

Size-exclusion chromatography of heparin oligosaccharides at high and low pressure

“…At pH > 1, the negatively charged GAGs eluted much earlier than at pH 1 (Fig. 5C), most likely due to electrostatic repulsion between the negatively charged GAGs and the matrix [26][27][28][29] or through intermolecular repulsion of heparin [30], despite sufficient content of counter ions in the mobile phase (see Section 3.9). Interestingly, this reduction in elution time continued also at pH higher than the pKa of the sulfate groups (pKa ≈ 0.5-1.5 [31]) suggesting that polarization of the matrix attributes additional repulsive forces to negatively charged GAGs.…”

“…Interaction of the analytes with the matrix may occur because most hydrophilic SEC matrices comprise residual negative charges or polarizeable functional groups [26][27][28][29]. As a consequence, addition of an electrolyte in the mobile phase may partially compensate this interaction through creation of a diffuse Coulomb layer.…”