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Rapid method for determining desmosine, isodesmosine, 5-hydroxylysine, tryptophan, lysinoalanine and the amino sugars in proteins and tissues

1986; Elsevier BV; Volume: 378; Linguagem: Inglês

10.1016/s0378-4347(00)80700-8

1872-812X

Constantinos G. Zarkadas, George C. Zarkadas, C.N. Karatzas, Ali D. Khalili, Quang D. Nguyen,

Bone and Dental Protein Studies

A rapid and sensitive chromatographic method is described for determining desmosine, isodesmosine, 5-hydroxylysine, tryptophan, lysinoalanine, glucosamine and galactosamine at picomole levels in protein and tissue hydrolysates. This method uses either an automated amino acid analyser with a 17.5 x 0.28 cm microcolumn packed with 6.0 ± 0.5 μm spherical resin, thermostated at 52°C, one buffer system (0.21 M sodium citrate, pH 5.125) and 3-nitrotyrosine as the internal standard, or conventional instruments using the same system but with larger diameter columns and resins (11.0 ± 1.0 μm). This method should be especially valuable for determining collagen and elastin in tissue hydrolysates from the amounts of 5-hydroxylysine, and desmosine or isodesmosine present, respectively, and for studying protein hydroxylation, glycosylation, cross-linking formation, and the turnover rates of collagen and elastin in normal and diseased tissues.