Determination of the glycosaminoglycan and collagen contents in tissue samples by high-resolution 1H NMR spectroscopy after DCl-induced hydrolysis.
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. (1)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 (1)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.
University of Leipzig , Faculty of Medicine, Institute of Medical Physics and Biophysics, Leipzig, Germany. riemer@uni-leipzig.de
Thomas
T
Riemer
Ariane
A
Nimptsch
Kathrin
K
Nimptsch
Jürgen
J
Schiller
D000116:Acetylgalactosamine / Q000737:chemistry; D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D002356:Cartilage / Q000737:chemistry; D002417:Cattle; D002809:Chondroitin Sulfates / Q000737:chemistry; D003094:Collagen / Q000737:chemistry*; D003903:Deuterium; D005109:Extracellular Matrix / Q000737:chemistry; D005944:Glucosamine / Q000737:chemistry; D006025:Glycosaminoglycans / Q000737:chemistry*; D006820:Hyaluronic Acid / Q000737:chemistry; D006851:Hydrochloric Acid / Q000737:chemistry*; D006868:Hydrolysis; D009682:Magnetic Resonance Spectroscopy; D013710:Tendons / Q000737:chemistry
D000596:Amino Acids; D006025:Glycosaminoglycans; D006820:Hyaluronic Acid; D002809:Chondroitin Sulfates; D003094:Collagen; D003903:Deuterium; D000116:Acetylgalactosamine; D005944:Glucosamine; D006851:Hydrochloric Acid