Silver staining of collagen type I after sodium dodecylsulphate polyacrylamide gel electrophoresis: effect of Maillard reaction.

Differences in the acidic silver staining, after sodium dodecylsulphate polyacrylamide gel electrophoresis, between the alpha 1 and alpha 2 collagen chains, as well as between rat-tail tendon and calf-skin collagen type I, were observed. The staining conditions at which the staining differences are both most expressed and reproducible were characterized. Age differences between staining of the alpha 1 CB6 fragment from young rats (2 and 12 months) and old rats (29 months) indicated that different susceptibilities of collagen species to the silver staining can be the result of different extents of some age-dependent post-translational modification, such as glycation.

Capillary zone electrophoresis: its applicability and potential in biochemical analysis.

Recent developments in capillary zone electrophoresis (CZE) are reviewed, starting with available instrumentation, a description of different operational modes and the most commonly used detection systems. Appropriate attention is paid to CZE-mass spectrometry coupling and coupling of electrophoretic and chromatographic procedures. The possibility of separating chiral molecules is also discussed.

Separation of elastin cross-links as phenylisothiocyanate derivatives.

A method has been developed for the separation and quantitation of desmosines in tissue samples. The tissue is treated with cold 10% trichloroacetic acid to remove collagen and hydrolysed in HCl vapours in sealed vials. Preseparation of desmosines from tissue acid hydrolysates is performed on a cellulose column, first eluted with n-butanol-acetic acid-water to wash out other amino acids and then with water to recover desmosines.

Change with age of UV absorbance and fluorescence of collagen and accumulation of epsilon-hexosyllysine in collagen from Wistar rats living on different food restriction regimes.

Accumulation of glycation products (as revealed by the thiobarbituric test and hexosyllysine assay) and the pigmented products (350 nm UV absorbance and 370ex/440em nm fluorescence) in aortal and skin collagen was investigated under the conditions of different nutritional regimes. Four groups of animals were tested: (1) ad libitum fed controls, (2) animals which were food restricted throughout their whole life (50% food intake), (3) animals fed ad libitum during their first year of life and then food restricted and (4) animals food restricted when young and fed ad libitum from the age of 1 year onwards. It was shown that all food-restricted animals showed lower levels of glycation and pigmentation products in collagen preparations from skin and aorta.

Separation and partial characterization of Maillard reaction products by capillary zone electrophoresis.

Capillary zone electrophoresis proved useful for separating small amounts of both charged and uncharged solutes that are otherwise difficult to analyse. A typical complex mixture that had previously resisted all analytical approaches, including reversed-phase separations, is the products arising from the reaction of free amino acids with aldehydic sugars (Maillard reaction products). By using capillary zone electrophoresis [untreated capillary 50 cm x 75 microns I.