LC-MS/MS Quantitation of Formaldehyde–Glutathione Conjugates as Biomarkers of Formaldehyde Exposure and Exposure-Induced Antioxidants: A New Look on an Old Topic

Humans are continuously exposed to formaldehyde via both endogenous and exogenous sources. Prolonged exposure to formaldehyde is associated with many human diseases, such as lung cancer and leukemia. The goal of this study is to develop biomarkers to measure formaldehyde exposure, which could be used to predict the risk of associated diseases. As glutathione (GSH) is well-known for its crucial role in the detoxification of a wide variety of xenobiotics, including formaldehyde, we rigorously quantitated in this study the conjugates formed when formaldehyde reacted with GSH using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) coupled with an isotope dilution method. The results showed for the first time that (S)-1-(((R)-2-amino-3-(carboxymethylamino)-3-oxopropylthio)methyl)-5-oxopyrrolidine-2-carboxylic acid (PGF) and thioproline-glycine (SPro-Gly) are major metabolites in both nonenzymatic reactions and formaldehyde-exposed human cells. In particular, over 35% of the formaldehyde from external sources was found to convert to SPro-Gly in the exposed cells. Interestingly, data showed that these exposure-induced adducts exhibited good antioxidative properties, which can protect cells from hydrogen peroxide mediated oxidative insult. It is anticipated that the findings of this study could shed light on developing PGF and SPro-Gly as dietary supplements and on the development of noninvasive methods to assess health risks associated with formaldehyde exposure.