Abstract
AbstractGlycation of collagen produces advanced glycation end products including pentosidine, glyoxal‐lysine dimer, and methylglyoxal‐lysine dimer. These products are markers for aging and associated with the development of several diseases. In this study, a rapid and simultaneous analytical method for pentosidine, glyoxal‐lysine dimer, and methylglyoxal‐lysine dimer was developed. The separation of advanced glycation end products was carried out on a silica hydride column using either acetonitrile or methanol in water with 0.1% (v/v) formic acid. A total run time of <5.5 min was achieved using isocratic elution and 11 min using gradient conditions. The gradient elution also allowed the rapid and simultaneous of collagen crosslinks and advanced glycation end products in one run. The eluted peaks were detected by mass spectrometry then the fragmentation was measured by tandem mass spectrometry to confirm their identity. The method developed herein was successfully applied to high‐performance liquid chromatography with fluorescence and ultraviolet detection. Although the ultraviolet detection sensitivity of glyoxal‐lysine and methylglyoxal‐lysine dimers is low, it is useful for preparative work for the collection of intact advanced glycation end‐products. This method was applied to several biological sample products and used to monitor the formation of glycation product in gelatin treated with glucose.
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