Size exclusion chromatography of polysaccharides with reverse phase liquid chromatography

He, Yue; Hou, Wenbo; Thompson, Melissa R; Holovics, Heidi J.; Hobson, Tracy; Jones, M. B.

doi:10.1016/j.chroma.2013.11.010

Cited by 18 publications

(9 citation statements)

References 20 publications

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“…The conjugation reaction is typically conducted in the presence of excess polysaccharide [4,9], requiring the removal of unreacted (free) polysaccharide from the final product [8]. A number of methods have been explored for purification of the conjugate including size exclusion [3,8], hydrophobic interaction [3,8], and reverse phase [10] chromatography, as well as liquid-liquid extraction [3] and ammonium sulfate precipitation [3]. Although the chromatographic methods do provide reasonably good selectivity, they have very low dynamic binding capacities due to the large size of the polysaccharides / conjugates.…”

Section: Introductionmentioning

confidence: 99%

Ultrafiltration behavior of bacterial polysaccharides used in vaccines

Hadidi

Buckley

Zydney

2015

Journal of Membrane Science

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Section: Introductionmentioning

confidence: 99%

Ultrafiltration behavior of bacterial polysaccharides used in vaccines

Hadidi

Buckley

Zydney

2015

Journal of Membrane Science

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“…Such a material seemed to be suitable for purification of polysaccharides by means of SEC because the hydrodynamic radius of polysaccharides with MW of 10–1000 kDa such as alginate, chitosan, pulluan, galactomannan, heparin and methyl cellulose, calculated from the Stokes–Einstein equation based on dynamic light scattering experiments is in the range of 70–340 Å [ 34 ]. Literature data [ 28 – 30 , 35 ] showed that bare and modified silica of larger pores (100–300 Å) can be used for SEC of polysaccharides in aqueous mode.…”

Section: Resultsmentioning

confidence: 99%

“…Earlier works [ 28 ] showed a possibility of fractioning dextrans of average MW 40 kDa into low and high fraction (25 and 70 kDa respectively) using mesoporous silica of particle size 200–500 micron. Bare silica was also investigated in HPLC mode (particle size 5 micron) [ 29 ] and SEC properties was showed for dextrans of MW between 5 and 100 kDa when pore size of the bed were 100–250 Å. Interestingly, a RP column with large pores (300 Å) was also suitable for purification and fractioning dextrans of MW 6–600 kDa if logMW difference was equal to 1 [ 30 ].…”

Section: Introductionmentioning

confidence: 99%

SEC Separation of Polysaccharides Using Macroporous Spherical Silica Gel as a Stationary Phase

et al. 2018

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Meso- and macroporous spherical silica gels of pore sizes in the range of 60–1000 Å and 40–75 µm particle size were investigated as a stationary phase for the separation and purification of polysaccharides and poly(ethylene glycols) (PEGs) of various MWs using an aqueous mobile phase. Sephadex and Bio-Gel were used for comparison as the most common stationary phases for similar purposes. The separation of dextrans of a mean MW = 31 kDa from small molecules (NaCl) was possible with SiO2 with a pore size of 60–300 Å, but the observed efficiencies of a column of the same size were lower comparing with Sephadex or Bio-Gel. In the case of oxidized alginic acid only SiO2 of the 60 Å pore size was suitable, while Sephadex, Bio-Gel and other investigated silicas were not efficient. Sephadex and 300–1000 Å SiO2 offered the possibility of dividing dextrans with MW within the range of 1 MDa–10 kDa into fractions of various MWs, while Bio-Gel and 60 Å SiO2 were not suitable. The investigated silica gels strongly adsorbed PEGs of MW 2–20 kDa. The amount adsorbed decreased with the increase of pore size and they were not useful as a stationary phase for this class of polymers. An advantage of SiO2 of the investigated particle size was a very low back pressure comparing with Sephadex. A considerably lower price of silica offers time- and cost-efficient separation of polysaccharides.Graphical Abstract

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“…A recent innovation has been the use of a reverse phase (RP) column for size-based separation of hydrophilic polysaccharides; the study included an investigation of stationary and mobile phase parameters and the coupling of RP-HPLC to MALLS (He et al 2014). A recent innovation has been the use of a reverse phase (RP) column for size-based separation of hydrophilic polysaccharides; the study included an investigation of stationary and mobile phase parameters and the coupling of RP-HPLC to MALLS (He et al 2014).…”

Section: Improved Characterization Of Solution Properties Of Carbohydmentioning

confidence: 99%