Quantitation by GC–MS of Methylglyoxal as a Marker in Anxiety-Related Studies

• Published in: Chromatographia 2013-05, Vol.76 (9-10), p.571-576
• Main Authors: Chen, Bud-Gen, Lin, Chia Han, Chen, Chinpiao, Hambsch, Boris, Chern, Chi-Liang
• Format: Article
• Language: English
• Subjects: Analytical Chemistry; Chemistry; Chemistry and Materials Science; Chromatography; Laboratory Medicine; Pharmacy; Proteomics; Short Communication
• ISSN: 0009-5893; 1612-1112
• DOI: 10.1007/s10337-013-2397-7

Methylglyoxal (MG) is present in the human system. In the cell, MG is degraded by glyoxalase 1 (Glo 1), an enzyme differentially expressed in several mouse models of anxiety-related behavior. It has also been associated with the formation of advanced glycation and lipoxidation end products, contributing to diabetes-related complications. Approaches developed for accurate quantitation of MG in this study include the synthesis of a deuterated MG analog to serve as an internal standard; skillful application of the standard addition protocol (with “reiteration”) to quantitate MG in “blank” samples, allowing the preparation of calibration standards using the same test specimen matrix system; and various pretreatment methods, including extraction and derivatization, to prepare complex brain tissue specimens for gas chromatography–mass spectrometry analysis. Using our method, brain MG levels in high- and low-anxiety-related behavior (HAB and LAB) mice were determined as 5.11 ± 0.89 and 13.05 ± 0.89 μg g −1 , respectively. These data supported the conclusion that MG level in brain is negatively correlated with anxiety in mouse models of anxiety-related studies, contributing to better understanding of the molecular mechanisms underlying reported observations.