Silica gel high-performance liquid chromatography for the determination of cross-links in elastin.
A high-performance liquid chromatographic (HPLC) procedure for the determination and preparation of desmosine (DES) and isodesmosine (IDE), the major cross-links of elastin, has been developed. Hydrolysates of elastin are treated with a SEP-PAK silica-gel small column and then separated on a silica-gel normal phase column using n-propanol-water-25% ammonia (9:3:0.06, v/v/v) followed by detection at 275 nm. The quantitative yields of DES and IDE are 7.98 +/- 0.47 and 6.44 +/- 0.51 (mean +/- S.D., n = 10) mg/g dry weight, respectively, in bovine ligamentum nuchae. Preparations of cross-links are carried out by means of a preparative column (240 mm x 10 mm), using the same solvent system.
Nakamura
F
F
Laboratory of Molecular Technology of Animal Products, Faculty of Agriculture, Tohoku University, Sendai, Japan.
Suyama
K
K
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D003432:Cross-Linking Reagents; D003895:Desmosine / Q000737:chemistry; D004549:Elastin / Q000737:chemistry*; D005260:Female; D007524:Isodesmosine / Q000737:chemistry; D008969:Molecular Sequence Data; D013056:Spectrophotometry, Ultraviolet
D003432:Cross-Linking Reagents; D003895:Desmosine; D004549:Elastin; D007524:Isodesmosine