Determining the molar mass of a plasma substitute succinylated gelatin by size exclusion chromatography-multi-angle laser light scattering, sedimentation equilibrium and conventional size exclusion chromatography.
The clinical effectiveness of succinylated gelatin as a plasma substitute depends strongly on its molar mass, determined conventionally by size exclusion chromatography (SEC). This study evaluates different SEC calibration standards in comparison with two independent "absolute" methods for determining the weight average molar mass (M(w)) of a succinylated gelatin sample. SEC calibrated using succinylated gelatin fractions correlated well with size exclusion chromatography-multi-angle laser light scattering (SEC-MALLS) and sedimentation equilibrium whereas SEC calibrated with unmodified gelatin, sodium polystyrene sulfonates or pullulans overestimated M(w) by over 20%. Universal calibration was equivocal. The problems associated with the preparation of succinylated gelatin fractions suggest that an absolute method such as SEC-MALLS may be a more suitable choice for determining the M(w) in succinylated gelatins.
Kaur
Manjit
M
Institute of Pharmaceutical Sciences Research, School of Pharmacy, University of Nottingham, UK.
Jumel
Kornelia
K
Hardie
Kim R
KR
Hardman
Andrea
A
Meadows
John
J
Melia
Colin D
CD
D002138:Calibration; D002850:Chromatography, Gel / Q000379:methods*; D005780:Gelatin / Q000737:chemistry*; D008027:Light; D010952:Plasma Substitutes / Q000737:chemistry*; D012542:Scattering, Radiation; D013386:Succinates / Q000737:chemistry*
D010952:Plasma Substitutes; D013386:Succinates; C060085:succinylated gelatin; D005780:Gelatin