Rapid method for the isolation of hydroxylysylpyridinoline and lysylpyridinoline from concentrated bone hydrolysates by liquid chromatographic techniques |
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Authors: | G. Lamprecht |
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Affiliation: | Institute for Analytical Chemistry, University of Vienna, Währinger Strasse 38, A-1090 Vienna, Austria |
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Abstract: | A rapid method for the isolation of hydroxylysylpyridinoline and lysylridinoline from bone by liquid chromatographic methods is described. Decalcified bone is hydrolysed in 7 M hydrochloric acid. After evaporation of the acid, the high molecular mass dark coloured degradation products are removed by adsorption on non-polar adsorbents. The pyridinolines are separated from the majority of the amino acids by adsorption on cellulose. Separation of HP and LP is performed either by cation-exchange chromatography or by reversed-phase ion-pari chromatography. The pyridinoline containing fractions are desalted by size-exclusion chromatography. The progress of the hydrolytic cleavage of collagen and the optimal parameters for purification and separation were examined. As a result the existing method allows the isolation of high amounts of pyridinolines with low amounts of adsorbents and chemical within a short time. |
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Keywords: | Bone hydrolysate Hydroxylysylpyridinoline Lysylpyridinoline Amino acids |
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