362
J. T . D I N G L E A N D M . W E B B
suitable salts. A micro-adaptation of this method by Gardell (1961),
in which glass-paper strips, dipped in cetylpyridinium chloride, were
used in place of the cellulose column, enabled 50 /xg amounts of each of
these compounds to be separated. Heparin and related sulphated
mucopolysaccharides have been separated by paper chromatography
with a mixture of ammonium formate buffer and isopropanol (65 : 35)
as solvent (Spolter and Marx, 1960), and recently Marsden and Kent
(1962) have developed a rapid micro-fractionation procedure, based
on the differential solubility of the cetylpyridinium complexes, for the
separation on paper of acidic mucopolysaccharides in the 5-70 /xg
range. This latter method appears potentially of great value in tissue
culture work.
Chromatography on modified cellulose (ECTEOLA) also separates
and gives excellent recovery of hyaluronic acid, chondroitin sulphate-A
and heparin (Ringertz and Reichard, 1959, 1960). As, however,
elution from these ion-exchange systems is most effective at low pH,
in the application of these methods degradative changes may occur in
certain mucopolysaccharides e.g. keratosulphate and JV-sulphated
mucopolysaccharides (Schiller et al., 1961).
3. Electrophoresis
Electrophoretic methods for the separation of mucopolysaccharide
fractions, either alone, or in combination with other procedures, have
been described by a number of authors, e.g. Gardell, Gordon and
Aqvist (1950), Schiller et al. (1954), Bollet (1958), Berenson (1958).
Separation of mucopolysaccharides by paper electrophoresis has been
described by Rienets (1953) and Berenson, Lumpkin and Shipp (1958)
among others. One difficulty in paper electrophoresis is the identification of carbohydrate fractions on a carbohydrate support. A quantitative method of estimation has been developed, however, by Kahan
(1959), and the use of glass fibre paper as described by Fuller and
Northcote (1956) may simplify the problems of identification.
G . C H E M I C A L E S T I M A T I O N OF
M U C O P O L Y S A C C H A R I D E C O N S T I T U E N T S
Until recently the characterization of isolated mucopolysaccharides
has rested on the determination of the contents of amino sugar, uronic
acid and sulphate. For the estimation of the amino sugars the most
commonly used method is that of Elson and Morgan (1933). The
specificity of this colorimetric procedure can be increased greatly if
the amino sugars are absorbed on a cation exchange resin (Boas, 1953)
and the impurities eluted with water, the amino sugars being eluted
J. T . D I N G L E A N D M . W E B B
suitable salts. A micro-adaptation of this method by Gardell (1961),
in which glass-paper strips, dipped in cetylpyridinium chloride, were
used in place of the cellulose column, enabled 50 /xg amounts of each of
these compounds to be separated. Heparin and related sulphated
mucopolysaccharides have been separated by paper chromatography
with a mixture of ammonium formate buffer and isopropanol (65 : 35)
as solvent (Spolter and Marx, 1960), and recently Marsden and Kent
(1962) have developed a rapid micro-fractionation procedure, based
on the differential solubility of the cetylpyridinium complexes, for the
separation on paper of acidic mucopolysaccharides in the 5-70 /xg
range. This latter method appears potentially of great value in tissue
culture work.
Chromatography on modified cellulose (ECTEOLA) also separates
and gives excellent recovery of hyaluronic acid, chondroitin sulphate-A
and heparin (Ringertz and Reichard, 1959, 1960). As, however,
elution from these ion-exchange systems is most effective at low pH,
in the application of these methods degradative changes may occur in
certain mucopolysaccharides e.g. keratosulphate and JV-sulphated
mucopolysaccharides (Schiller et al., 1961).
3. Electrophoresis
Electrophoretic methods for the separation of mucopolysaccharide
fractions, either alone, or in combination with other procedures, have
been described by a number of authors, e.g. Gardell, Gordon and
Aqvist (1950), Schiller et al. (1954), Bollet (1958), Berenson (1958).
Separation of mucopolysaccharides by paper electrophoresis has been
described by Rienets (1953) and Berenson, Lumpkin and Shipp (1958)
among others. One difficulty in paper electrophoresis is the identification of carbohydrate fractions on a carbohydrate support. A quantitative method of estimation has been developed, however, by Kahan
(1959), and the use of glass fibre paper as described by Fuller and
Northcote (1956) may simplify the problems of identification.
G . C H E M I C A L E S T I M A T I O N OF
M U C O P O L Y S A C C H A R I D E C O N S T I T U E N T S
Until recently the characterization of isolated mucopolysaccharides
has rested on the determination of the contents of amino sugar, uronic
acid and sulphate. For the estimation of the amino sugars the most
commonly used method is that of Elson and Morgan (1933). The
specificity of this colorimetric procedure can be increased greatly if
the amino sugars are absorbed on a cation exchange resin (Boas, 1953)
and the impurities eluted with water, the amino sugars being eluted