Simultaneous determination of the advanced glycation end product N (epsilon)-carboxymethyllysine and its precursor, lysine, in exhaled breath condensate using isotope-dilution-hydrophilic-interaction liquid chromatography coupled to tandem mass spectromet

Abstract

Analysis of biomarkers in exhaled breath condensate (EBC) is a non-invasive method for investigating the effects of different diseases or exposures, on the lungs and airways. N(epsilon)-carboxymethyllysine (CML) is an important biomarker of advanced glycation end products (AGEs). A method has been developed for simultaneous determination of CML and its precursor, the amino acid lysine, in exhaled breath condensate (EBC). After addition of labelled internal standards (d-4-CML; d-4-lysine), the EBC was concentrated by freeze-drying. Separation and detection of the analytes were performed by hydrophilic-ion liquid chromatography coupled with tandem mass-spectrometric detection (HILIC-MS-MS). The limits of quantification were 10 pg mL(-1) EBC and 0.5 ng mL(-1) EBC for CML and lysine, respectively. The relative standard deviation of the within-series precision was between 2.8 and 7.8% at spiked concentrations between 40 and 200 pg mL(-1) for CML and between 6 and 20 ng mL(-1) for lysine. Accuracy for the analytes ranged between 89.5 and 133%. The method was used for the analysis of EBC samples from ten healthy persons from the general population and ten persons receiving dialysis. CML and lysine were detected in all EBC samples with median values of 19 pg mL(-1) CML and 11.9 ng mL(-1) lysine in EBC of healthy persons and 25 pg mL(-1) CML and 9.5 ng mL(-1) lysine in EBC of dialysis patients.