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Frontiers of Environmental Science & Engineering

ISSN 2095-2201

ISSN 2095-221X(Online)

CN 10-1013/X

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Front. Environ. Sci. Eng.    2017, Vol. 11 Issue (2) : 16    https://doi.org/10.1007/s11783-017-0922-x
RESEARCH ARTICLE
Tetra-detector size exclusion chromatography characterization of molecular and solution properties of soluble microbial polysaccharides from an anaerobic membrane bioreactor
Qingbin Guo,Sheng Chang()
School of Engineering, University of Guelph, Guelph, Ontario, N1G 2W1, Canada
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Abstract

Tetra-detector HPSEC was evaluated for the SMP characterization

Molecular weight and intrinsic viscosity of the SMP were characterized

Specific viscosity and osmotic pressure of the SMP solution were studied

Approach to analyze the concentration polarization of the SMP was discussed

Characterization of the molecular properties of soluble microbial products (SMP) is critical for understanding the membrane filtration and fouling mechanisms in anaerobic and aerobic membrane bioreactors (AnMBR & MBR). In this study, the distributions of the absolute molecular weight and intrinsic viscosity of SMP polysaccharides from an AnMBR were effectively determined by a high performance size exclusion chromatography (HPSEC) that was coupled with the refractive index (RI), diode array UV (DAUV), right and low angle light scattering (LS), and viscometer (Vis) detectors. Based on the tetra-detector HPSEC determined absolute molecular weights and intrinsic viscosity, a universal calibration relationship for the SMP polysaccharides was developed and the molecular conformations, average molecular weights, and hydrodynamic sizes of the SMP polysaccharides were also explored. Two factors which can be derived from the tetra-detector HPSEC analysis were proposed for the characterization of the viscous and osmotic pressure properties of the SMP polysaccharides. In addition, it was also extrapolated how to analyze the resistance characteristics of the concentration polarization layers formed in membrane filtration based on the molecular properties determined by the tetra-detector HPSEC analysis.
Keywords Soluble microbial product      Extracellular polymeric substance      Membrane bioreactor      Membrane fouling      Molecular weight      Intrinsic viscosity     
Corresponding Author(s): Sheng Chang   
Issue Date: 07 April 2017
 Cite this article:   
Qingbin Guo,Sheng Chang. Tetra-detector size exclusion chromatography characterization of molecular and solution properties of soluble microbial polysaccharides from an anaerobic membrane bioreactor[J]. Front. Environ. Sci. Eng., 2017, 11(2): 16.
 URL:  
https://academic.hep.com.cn/fese/EN/10.1007/s11783-017-0922-x
https://academic.hep.com.cn/fese/EN/Y2017/V11/I2/16
sample
name		 operation days reactor operation conditions
R1 0 before reactor start (10 kPa 100 kDa filtration)
R2 56 5 gCOD·L·d−1, no sludge wasting (10 kPa 100 kDa filtration)
R3 71 7.5 gCOD·L·d−1, no sludge wasting (10 kPa 100 kDa filtration)
R4 94 10 gCOD·L·d−1, with sludge wasting (10 kPa 100 kDa filtration)
Tab.1  SMP samples prepared for the tetra-detector HPSEC analysis
Fig.1  Tetra-detector chromatograms of soluble microbial products (SMP) for the sample R4
Fig.2  (a) Universal calibration curves of SMP polysaccharides. (b) log(intrinsic viscosity)-log (molecular weight) relations of SMP polysaccharides
samples RI Peak EV /mL a values b values R2
R1 23.224 −0.58 19.2 0.9932
R2 23.036 −0.52 17.7 0.9982
R3 22.906 −0.58 19.2 0.9828
R4 22.58 −0.54 18.1 0.9897
average values ? −0.55 18.6 ?
standard deviations ? 0.03 0.77 ?
Tab.2  Linear fitting constants (a and b) of the Log (intrinsic viscosity x molecular weight) and elution volume (mL) relation: Log([h]M) = aEV+ b, where [h] is the intrinsic viscosity (dl·g−1), M is the molecular weight (Da), and EV is the elution volume (mL).
MW<105.5 Da 105.5 <MW<106 Da MW>106 Da
sample Mw /KDa PDI [h]/(d·g −1) F/% α/logK Mw/KDa PDI [h]/(d·g −1) F/% α/logK Mw /KDa PDI [h]/(d·g−1) F/% α/logK
R1 181.5 1.18 2.10 61.8 1.49/−7.75 562.3 1.11 5.83 20.2 0.45/-1.99 3266.4 1.62 21.04 17.9 0.67/-3.53
R2 143.5 1.18 1.48 68.2 1.56/−7.70 575.5 1.11 5.92 17.9 0.44/-1.78 2415.9 1.3 15.82 14.0 0.74/-3.58
R3 185.5 1.24 2.16 45.3 1.16/−5.89 549.4 1.11 5.75 27.4 0.62/-2.87 3159.8 1.36 20.39 27.2 0.85/-4.32
R4 181.2 1.2 2.09 55.5 1.78/−8.91 534.3 1.1 5.65 27.7 0.69/-3.06 2069.8 1.12 13.67 16.7 1.03/-5.12
mean 177.1 1.21 2.03 57.7 1.50/−7.56 545.5 1.11 5.73 23.3 0.55/-2.43 2690.0 1.31 17.49 18.9 0.82/-4.14
stdev 18.5 0.04 0.31 9.7 0.25/1.08 20.8 0.00 0.14 5.0 0.13/0.55 614.8 0.19 3.79 5.7 0.16/0.65
Tab.3  Weight average molecular weight (Mw), polydispersity index (PDI) values, intrinsic viscosity ([h]), mass fraction (F) and Mark-Houwink parameters (a and logK) of the SMP polysaccharides for different molecular weight ranges
MW<105.5 Da 105.5 <MW<106 Da MW>106 Da
sample Rg /nm Rh/nm ρ/(Rg/Rh) Rg /nm Rh/nm ρ/(Rg/Rh) Rg /nm Rh/nm ρ/(Rg/Rh)
R1 26.1 18.2 1.43 45.8 37.3 1.23 151.2 102.9 1.47
R2 21.5 15.0 1.43 46.4 37.8 1.23 124.3 84.6 1.47
R3 26.6 18.5 1.44 45.2 36.9 1.22 148.0 100.7 1.47
R4 26.1 18.2 1.43 44.6 36.3 1.23 112.5 76.6 1.47
mean 25.1 17.5 1.43 45.5 37.1 1.23 134.0 91.2 1.47
stdev 2.1 1.4 0.00 0.7 0.5 0.00 16.2 11.0 0.00
Tab.4  Average radius of gyration (Rg), viscometric radius (Rh) and conformational parameter (r) of the SMP polysaccharides for different molecular weight ranges
Fig.3  (a) SFi[h]i values of the SMP polysaccharides. (b) Relationships between osmotic pressure and concentration for the SMP polysaccharides
Fig.4  Approach to solve the concentration polarization problem (K is the permeability, Rg is the radius of gyration, f is the volume fraction of polymer, f* is the overlap concentration of polymer, D is the diffusivity of polymer molecule, η0 is the solvent viscosity, p is the osmotic pressure, c is the polymer mass concentration, J is the flux, Rc is the resistance of concentration layer, Dx is the thickness of the concentration polarization layer, and sub index i represents a thin concentration polarization layer i)
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