Rapid method for the isolation of hydroxylysylpyridinoline and lysylpyridinoline from concentrated bone hydrolysates by liquid chromatographic techniques.
A rapid method for the isolation of hydroxylysylpyridinoline and lysylyridinoline 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 and 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-pair 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 chemicals within a short time.
Lamprecht
G
G
Institute for Analytical Chemistry, University of Vienna, Austria.
D000596:Amino Acids / Q000302:isolation & purification*; D001842:Bone and Bones / Q000737:chemistry*; D002853:Chromatography, Liquid; D006868:Hydrolysis
D000596:Amino Acids; C015484:pyridinoline; C036020:deoxypyridinoline