The Use of Solid Phase Extraction Cartridges as a Pre-Fractionation Step in the Quantisation of Intermolecular Collagen Crosslinks and Advanced Glycation End-Products

Abstract

A range of solid phase extraction (SPE) sorbents was investigated to determine their suitability as a pre-fractionation step during the quantitation of collagen crosslinks and advanced glycation end products by reverse phase high performance liquid chromatography (RP HPLC).

Propylsulphonic acid (PRS) SPE cartridges were shown to be the most suitable sorbent type by testing for resolution of specific amino acids using both inorganic salt buffers and volatile organic solvents.

The loading capacity of the PRS-SPE cartridges was determined and the efficacy of the cartridges evaluated by comparison with previously published methods using fibrous cellulose and SP Sepharose.

PRS-SPE cartridges provided a superior, rapid, cost effective alternative to either of these pre-fractionation steps giving a higher recovery of both the mature collagen crosslink, pyridinoline, and the advanced glycation end-product crosslink, pentosidine.