Multicomponent analysis by off-line combination of synchronous fluorescence spectroscopy and capillary electrophoresis of collagen glycation adducts

Capillary electrophoresis separation and synchronous fluorescence spectral detection was used off-line to reveal the nature of fluorescent adducts for med in vivo in the collagen molecule and their distribution in the molecule . It was shown that by using the Delta lambda in the area of the Stokes shi ft for the analyzed entities (similar to 10 nm for pentosidine, 4,5(E)-epox y-2(E)-heptenal and 4,5(E)-epoxy-2(E)-decenal lysine adducts) a distinct pr ofile of spectral bands can be obtained allowing for differentiation of the several entities involved. In combination with capillary electrophoretic s eparation of the CNBr peptides the location of individual adducts was possi ble: while pentosidine land, perhaps, pentosidine related compounds K-1-K-4 ) is found in the large alpha(1)(I)CB6 and alpha(2)(I)CB3,5 peptides along with a complete set of the other fluorescent adducts, low-molecular-mass pe ptides originating from the terminal region of the molecule are devoid of a ny fluorescence. All other parts of the molecule possess synchronous fluore scence profiles corresponding to the intact molecule except that they are d evoid of pentosidine. The results indicate random distribution of fluoresce nt adducts in the collagen molecule and, in a broader context, indicate the usefulness of multicomponent analysis by means of combining synchronous lu minescence spectra and capillary electrophoresis. (C) 1999 Elsevier Science B.V. All rights reserved.