Abstract
Recently, desmosine (D) and isodesmosine (ID) in elastin hydrolysates have been analyzed by high-performance liquid chromatography (HPLC). However, it is difficult to resolve ID and ID by these procedures and the regular HPLC procedures themselves are time-consuming. For these reasons, a rapid and simple determination of D anb ID dy HPLC was developed. D and ID were separated on an HPLC column (Nucleosil C-18) with a mobile phase of 0.1 M CH3SO3H(pH 2.0)/CH3CN=90/10 containing 6 mM heptane sulfonate or 90/5 containing 2 mM heptane sulfonate. We applied the method for the analysis of D and ID contained in the elatstin hydrolysates of bovine ligamentum nuchae, smooth muscle culture medium, and rat aorta. Results of the analysis of the ligamentum nuchae and the medium agreed with those obtained by automatic amino acid analysis.
The method has advantages over other types of analysis for D and ID. It is accurate, reproducible, and simple and allows rapid analysis. Furthermore, its sensitivity is such that elastin from small amount of tissue that containslow concentrations of elastin can be quantitated.
The determination of D and ID in tissues or cell culture medium containing elastin may assume the same significance as the determination of collagen based on its hydroxyproline content.
