A number of techniques are available for fractionation and isolation of acid mucopolysaccharides.
In one procedure, AMPS* * is precipitated from an aqueous solution containing different metal ions (Ca, Mg, Ba) with organic solventslss. Ion exchange chromatography with Dowex 394, DEAEG or ECTEOLA celluloseO have been used by other investigators. The complex formed between polyanions and quaternary ammonium salts, described by SCOTT 78 has since been utilized as the basis for the isolation of individual fractions by several investigatorso-11. It has been our experience that none of these methods provides sharp separations.
ANTONOFOULOS and co-workers12 recommended a method combining the SCOTT procedure with column chromatography. This provided clean separations of HA, the CS group and Hep, but wasnot capable of distinguishing members of the chondroitin sulfate group on Dowex I, DEAE or ECTEOLA columnsl+l6.
To permit identification of members of the chondroitin sulfate group in investigations of arterial AMPS, a two-step procedure was adopted. The first step served to separate HA, HMS, and the CS group by elution from a Dowex r column4~i6 or the CP-AMPS complexes from a cellulose columnlo, in the second step, the CS group was subjected to alcohol fractionation on a Hyflo supercel column, an adaptation of KAPLAN AND MEYER'S technique 17. It proved feasible to combine these two steps on a single cellulose column for the sharp separation of individual components from the mixture in isolated aortic AMPS.
MATERIALS CIFONELLI, Chicigo University;
CS-C (Kakenaku-Kako Co. Ltd.,