Employment of gas-liquid chromatography for the analysis of collagen amino acids in biopsy tissue

Abstract

In this paper, gas-liquid chromatography, adapted for the determination of collagen amino acids, is described. This technique was attractive for its sensitivity in that only a small amount of protein such as in 0.5 mg of tissue, especially as obtained from biopsy tissue, was needed for the separation and determination of proline (Pro), 4-hydroxyproline (4-Hyp), 3-hydroxyproline (3-Hyp), lysine (Lys), hydroxylysine (Hyl) and epsilon-hydroxy-norleucine (epsilon-PH-Norleu), the characteristic amino acids of collagen. Thus, without purification of collagen, by determining the ratio Hyl/4-Hyp and 4-Hyp/Pro it was possible to determine some anomalies in the collagen content of biopsy tissue (skin or liver). The ratio Hyl/4-Hyp allows an estimation of the lack of hydroxylation of polypeptidic lysine as in the Ehlers-Danlos syndrome type VI; and the ratio 4-Hyp/Pro allows measurement of variations in collagen content in relation to protein, especially in the liver, as in alcoholic cirrhosis.