Determination of the Glycosaminoglycan
and Collagen
Contents in Tissue Samples by High-Resolution <sup>1</sup>H NMR Spectroscopy
after DCl-Induced Hydrolysis
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Abstract
The determination of the collagen and glycosaminoglycan
(GAG) contents
of native and particularly bioengineered tissues is of considerable
interest because the collagen-to-GAG ratio determines the water content
of the tissue, which is crucial regarding its mechanical properties. <sup>1</sup>H NMR spectroscopy subsequent to the hydrolysis of the sample
by aqueous 6 M DCl at 353 K is used to determine the GAG and collagen
contents simultaneously. Under these strongly acidic conditions the
biopolymers of the extracellular matrix, collagen, and GAG are fragmented
into their individual monomers, that is, free amino acids from collagen
and monosaccharides from the polymer repeat units of GAGs. The amino
acid amount can be easily determined in the presence of an internal
standard by <sup>1</sup>H NMR spectroscopy because amino acids proved
to be stable under acidic conditions. The carbohydrates are subject
to charring in the presence of concentrated DCl, but glucosamine and
galactosamine were found to be sufficiently stable for quantification
under the chosen conditions