MOLECULARDIMENSIONANDELUTIONVOLUMIE172.2HydrodynamicVolumeConceptSmall molecularAS=0A.dimensionKsec = 1CV,VA =V,V9RIntermediateAS<0B.SKsec =0.5VoV,V<V,LargemolecularC.AS<<0dimensionKsec→0V.V,VA=VoFig.2.5.Thermodynamic representation of the SEC process.Elution of:A totally permeatedrnolecules; Bpartiallyexcludedmolecules;Ctotallyexcluded molecules
MOLECULAR DIMENSION AND ELUTION VOLUME Small molecular dimension Intermediate Large molecular dimension
SCHEMATIC REPRESENTATION OFA SIZE-EXCLUSIONCHROMATOGRAPHYSET-UPCelladicaricamentoPOMPAValvolaa4vieTamponealcalinoColonnadipermeazione di gelREGISTRATORERIVELATORE
SCHEMATIC REPRESENTATION OF A SIZE-EXCLUSION CHROMATOGRAPHY SET-UP
GELPERMEATIONINNa,B,OzH9.20.02M0.1MLow pressureIIA.HA dissolved at pH 9.2B.Titrated topH6.0with acetic acidC.topH4.5withaceticacidD.to pH3.5 with acetic acidoeE. to pH 2.0 with acetic acidF.BacktitratedtopH3.5withKOHG.topH4.5withKOHH.topH6.0withKOHI.topH8.5withKOHJ.androtoevaporatedatpH8.56060120120VoVoPiccolo,Nardi,Concheri(1996).European Journal of SoilScience,47:319-328
GEL PERMEATION IN Na4 B2 O7 P H 9.2 Low pressure 0.02M 0.1M A. HA dissolved at pH 9.2 B. Titrated to pH 6.0 with acetic acid C. to pH 4.5 with acetic acid D. to pH 3.5 with acetic acid E. to pH 2.0 with acetic acid F. Backtitrated to pH 3.5 with KOH G. to pH 4.5 with KOH H. to pH 6.0 with KOH I. to pH 8.5 with KOH J. and rotoevaporated at pH 8.5 Piccolo, Nardi, Concheri (1996). European Journal of Soil Science, 47:319-328
HPSECelutioninNaNO,0.05MpH770-Monocarboxvlic acids65A=HA(volcanic soil)pH760AMw=32961B55cDB=topH3.5withHCl50-LHCINMw=-13.7%4540-C-withHCOOH35△MW=20.8%30D=with CH,COOH25-AMw=-29.8%20-E=withCH.CH,COOH15104Mw=-5.7%CH,COOH5F= with CH,(CH2),COOHHCOOH0151827303391122124AMw=+2.3%VoElutionvolume/mlPiccolo,Conte,Cozzolino(1999).European Journal of Soil Science,50:687-694
Monocarboxylic acids A= HA (volcanic soil) pH 7 Mw=32961 B= to pH 3.5 with HCl ΔMw=-13.7% C=with HCOOH ΔMw=-20.8% D=with CH3 COOH ΔMw=-29.8% E=with CH3 CH2 COOH ΔMw=-5.7% F= with CH3 (CH2 )2 COOH ΔMw=+2.3% CH3 COOH HCOOH HCl Piccolo, Conte, Cozzolino (1999). European Journal of Soil Science, 50:687-694 HPSEC elution in NaNO3 0.05 M pH 7
HPSECelutioninNaNO,0.05MpH7Sulphonic acids60-A=HA(oxidizedcoal)55-ABpH7c50-DMw-941845-40-B=topH3.5withHClSI35-AMw=-8.4%30-C=to pH3.5with25methansulphonic acid20-AMw=-27.0%15-10-D=topH3.5with5-benzensulphonicacid0302112151824279AMw=-28.7%ELUTION VOLUME(mL)Cozzolino,Conte,Piccolo(2001)Soil Biology and Biochemistry,33:563-571
A= HA (oxidized coal) pH 7 Mw=9418 B=to pH 3.5 with HCl ΔMw=-8.4% C= to pH 3.5 with methansulphonic acid, ΔMw=-27.0% D= to pH 3.5 with benzensulphonic acid ΔMw=-28.7% HPSEC elution in NaNO3 0.05 M pH 7 Sulphonic acids Cozzolino, Conte,Piccolo (2001) Soil Biology and Biochemistry, 33:563-571