Reactive carbonyl compounds (RCCs) are ubiquitous in the environment and are generated endogenously as a result of various physiological and pathological processes. These compounds can react with biological molecules inducing deleterious processes believed to be at the basis of their toxic effects. Several of these compounds are implicated in neurotoxic processes, aging disorders, and cancer. Therefore, a method characterizing exposures to these chemicals will provide insights into how they may influence overall health and contribute to disease pathogenesis. Here, we have developed a high resolution accurate mass (HRAM) screening strategy allowing simultaneous identification and relative quantitation of DNPH-derivatized carbonyls in human biological fluids. The screening strategy involves the diagnostic neutral loss of hydroxyl radical triggering MS 3 fragmentation, which is only observed in positive ionization mode of DNPH-derivatized carbonyls. Unique fragmentation pathways were used to develop a classification scheme for characterizing known and unanticipated/unknown carbonyl compounds present in saliva. Furthermore, a relative quantitation strategy was implemented to assess variations in the levels of carbonyl compounds before and after exposure using deuterated d 3 -DNPH. This relative quantitation method was tested on human samples before and after exposure to specific amounts of alcohol. The nano-electrospray ionization (nano-ESI) in positive mode afforded excellent sensitivity with detection limits on-column in the high-attomole levels. To the best of our knowledge, this is the first report of a method using HRAM neutral loss screening of carbonyl compounds. In addition, the method allows simultaneous characterization and relative quantitation of DNPH-derivatized compounds using nano-ESI in positive mode. [Figure not available: see fulltext.]
|Original language||English (US)|
|Number of pages||11|
|Journal||Journal of the American Society for Mass Spectrometry|
|State||Published - Apr 1 2017|
Bibliographical noteFunding Information:
The authors would like to thank the volunteers of the research study "Acetaldehyde-derived DNA adducts for the investigation of alcohol hangover" at the University of Minnesota and Robert Carlson for editorial assistance. This work was supported by NIOSH-funded MCOHS ERC Pilot Research Training Program (OH008434). The contents of this article are solely the responsibility of the authors and do not necessarily represent the official view of the National Institute for Occupational Safety and Health, Centers for Disease Control and Prevention, or other associated entities. Mass spectrometry was carried out in the Analytical Biochemistry Shared Resource of the Masonic Cancer Center, University of Minnesota, funded in part by Cancer Center Support Grant CA-077598 and S10 RR-024618 (Shared Instrumentation Grant).
© 2016, American Society for Mass Spectrometry.
- Aldehydes/carbonyl compounds
- High resolution accurate mass data-dependent MS neutral loss screening
- Relative quantitation