Characterization of Marine-Surface-Dissolved Organic Matter via Amino Acid Enantiomers and Its Implications Based on Diel and Seasonal Observations

Due to the essential roles of dissolved organic matter (DOM) in both microbiol food loop and marine carbon cycling, changes in marine DOM composition have an important impact on the marine ecosystem and carbon cycling. In October 2014 and June 2015, two field investigations for the DOM in the upper 200 m were conducted in the slope region of the northern South China Sea to characterize the DOM composition via amino acid enantiomers. In June, our sampling locations were under upwelling impact induced by an eddy-pair event, whereas in October there were no eddies. High-frequency sampling (a few hours interval) over 24 h reveals that the variability of the amino acid carbon yield (min. 0.2%) and the D/L alanine ratio (min. 0.02) is larger than its corresponding analytical and propagated errors, suggesting solid short-term changes for these two molecular-based indicators. Section samples from June showed a lower D/L alanine ratio (0.43 vs. 0.53) and a GABA mol% (1.0% vs. 1.6%) relative to the section samples from October, suggesting that DOM in June is more fresh (less degraded) compared to that in October. A higher serine mol% (19.5% vs. 13.2%) and lower D/L serine ratio (0.06 vs. 0.24) from the diel observation in June relative to October further indicates that phytoplankton, rather than bacteria, plays an more important role in DOM composition alternation. This is consistent with the higher phytoplankton biomass found in June, promoted by the eddy-pair.

Simultaneous Measurement of Amino Acid Enantiomers in Aged Mouse Brain Samples by LC/MS/MS Combined with Derivatization Using Nα-(5-Fluoro-2,4-dinitrophenyl)-l-leucinamide (l-FDLA)

d-amino acids have distinct roles from their l-enantiomer. In particular, some d-amino acids function as agonists or antagonists of neuronal receptors and are involved in higher brain functions. Thus, it is important to precisely measure the levels of these amino acid enantiomers in cells and tissues. Various quantification methods have been developed for measurements of chiral amino acids. However, each method has advantages and disadvantages. Additionally, measuring the amino acid enantiomers in crude biological samples requires a higher selectivity. In this study, we developed a quantification method for amino acid enantiomers using derivatization with Nα-(5-Fluoro-2,4-dinitrophenyl)-l-leucinamide (l-FDLA) followed by liquid chromatography–tandem mass spectrometry (LC/MS/MS) with a conventional reversed-phase column. We simultaneously identified 10 chiral amino acids. Furthermore, we applied this method to investigate murine tissue samples and examined the effect of aging on the amino acid levels in aged brain regions. We found that aging decreased the levels of both d-serine and d-aspartate in the hippocampus. In addition, d-Phenylalanine in the thalamus significantly increased with age. In conclusion, our method is suitable for the quantification of the d-amino acids in crude biological samples and may contribute to elucidating the biological roles of chiral amino acids.

Efficient Synthesis of New Fluorinated β-Amino Acid Enantiomers through Lipase-Catalyzed Hydrolysis

An efficient and novel enzymatic method has been developed for the synthesis of β-fluorophenyl-substituted β-amino acid enantiomers through lipase PSIM (Burkholderia cepasia) catalyzed hydrolysis of racemic β-amino carboxylic ester hydrochloride salts 3a–e in iPr2O at 45 °C in the presence of Et3N and H2O. Adequate analytical methods were developed for the enantio-separation of racemic β-amino carboxylic ester hydrochlorides 3a–e and β-amino acids 2a–e. Preparative-scale resolutions furnished unreacted amino esters (R)-4a–e and product amino acids (S)-5a–e with excellent ee values (≥99%) and good chemical yields (>48%).