Size and shape of soil humic acids estimated by viscosity and molecular weight

Ultrafiltration fractions of three soil humic acids were characterized by viscometry and high performance size-exclusion chromatography (HPSEC) in order to estimate shapes and hydrodynamic sizes. Intrinsic viscosities under given solute/solvent/temperature conditions were obtained by extrapolating the concentration dependence of reduced viscosities to zero concentration. Molecular mass (weight average molecular weight (M¯w) and number average molecular weight (M¯n)) and hydrodynamic radius (RH) were determined by HPSEC using pullulan as calibrant. Values of M¯w and M¯n ranged from 15 to 118×103 and from 9 to 50×103 (g mol−1), respectively. Polydispersity, as indicated by M¯w/M¯n, increased with increasing filter size from 1.5 to 2.4. The hydrodynamic radii (RH) ranged between 2.2 and 6.4 nm. For each humic acid, M¯w and [η] were related. Mark-Houwink coefficients calculated on the basis of the M¯w−[η] relationships suggested restricted flexible chains for two of the humic acids and a branched structure for the third humic acid. Those structures probably behave as hydrated sphere colloids in a good solvent. Hydrodynamic radii of fractions calculated from [η] using Einstein's equation, which is applicable to hydrated sphere colloids, ranged from 2.2 to 7.1 nm. These dimensions are fit to the size of nanospaces on and between clay minerals and micropores in soil particle aggregates. On the other hand, the good agreement of RH values obtained by applying Einstein's equation with those directly determined by HPSEC suggests that pullulan is a suitable calibrant for estimation of molecular mass and size of humic acids by HPSEC.