Alternative electrolyte solutions for untargeted breath metabolomics using secondary‐electrospray ionization high‐resolution mass spectrometry

Rationale Secondary‐electrospray ionization (SESI) coupled with high‐resolution mass spectrometry is a powerful tool for the discovery of biomarkers in exhaled breath. A primary electrospray consisting of aqueous formic acid (FA) is currently used to charge the volatile organic compounds in breath. To investigate whether alternate electrospray compositions could enable different metabolite coverage and sensitivities, the electrospray dopants NaI and AgNO3 were tested. Methods In a proof‐of‐principle manner, the exhaled breath of one subject was analyzed repeatedly with different electrospray solutions and with the help of a spectral stitching technique. Capillary diameter and position were optimized to achieve proper detection of exhaled breath. The detected features were then compared using formula annotation. Using an evaporation‐based gas standard system, the signal response of the different solutions was probed. Results Principal component analysis revealed a substantial difference in features detected with AgNO3. With silver, more sulfur‐containing features and more unsaturated hydrocarbon compounds were detected. Furthermore, more primary amines were potentially ionized, as indicated by van Krewelen diagrams. In total, twice as many features were unique to AgNO3 than for other electrospray dopants. Using gas standards at known concentrations, the high sensitivity of FA as a dopant was demonstrated but also indicated alternate sensitivities of the other electrospray solutions. Conclusions This work demonstrated the potential of AgNO3 as a complementary dopant for further biomarker discovery in SESI‐based breath analysis.