Determination of molecular weight distribution of acid mucopolysaccharides by Sephadex gel filtration

A method has been developed for the determination of molecular weight distribution of AMPS. Chondroitin sulfate A was fractionated by gel filtration on Sephadex G-200 into bands of narrow distribution of molecular weights. The molecular weights of the AMPS from bands widely separated were determined by a combined diffusion-sedimentation coefficient method. Aliquots from the same AMPS preparations were also subjected to gel filtration on a 1.06 (i.d.) 86.0 cm Sephadex G-200 column. It was found that a linear relationship exists between elution volume and log of molecular weight of the respective AMPS. Chondroitin sulfate B, chondroitin sulfate C, and heparitin sulfate follow the same relationship. Elution diagrams of the urinary AMPS of the three main variants of the Hurler syndrome are characteristically distinct from each other and they are also different from the elution diagrams of AMPS obtained from normal subjects. Heparitin sulfate and a portion of chondroitin sulfate B from Hurler patients are markedly degraded.