Determination of Pyridinolines in Bovine Collagenous Tissues

A new and sensitive method is described for the determination of the trivalent collagen cross-linkers hydroxylysylpyridinoline (HP) and lysylpyridinoline (LP) in intramuscular and other connective tissues. The procedure includes hydrolysis of the isolated connective tissue, filtration and phosphocellulose chromatography of the crude hydrolysates, and reversed-phase ion-paired high-performance liquid chromatography (RP-HPLC), using sodium octanesulfonic acid as ion-pairing agent. Detection is done by measuring the natural fluorescence of pyridinolines with a spectrofluorometer. The method has been tested by determining pyridinolines in several bovine connective tissues varying in collagen content