abstract
1D, and 2D 800 MHz high-resolution nuclear magnetic resonance spectroscopy of human amniotic fluid (HAF) enabled the identification of similar to 50 metabolites. In addition, liquid chromatography-NMR and diffusion ordered spectroscopy (DOSY) allowed signal overlap to be reduced and the characterization of higher molecular weight (M-w) components, respectively. Indeed, the DOSY spectrum of,a M-w >10 kDa HAF fraction enabled three protein families, differing in average Mw, to be detected and may therefore be of potential value in the study of disorderrelated variations in HAF protein profiles. The effects of freeze-drying, storage at -20 or -70 degrees C, and freeze-thawing cycles on HAF compositional stability were investigated, as well as stability at room temperature (to account for overnight data acquisition runs). These data are the basis for establishing statistically validated correlations between HAF NMR data and any physiological disorders of the fetus/mother. Freeze-drying caused signal loss for urea, ethanol, and compounds resonating at 2.22 and 1.17 ppm. Storage at -70 degrees C or lower is recommended since only minor compositional changes were observed, affecting mainly acetate and pyruvate. Freeze-thaw cycles did not cause significant compositional changes, and room-temperature stability studies indicated a 4-5 h maximum period of handling/ acquisition time to ensure HAF stability.
keywords
MAGNETIC-RESONANCE-SPECTROSCOPY; RESOLUTION H-1-NMR SPECTROSCOPY; DIFFUSION-ORDERED SPECTROSCOPY; P-31 NMR; PLASMA; H-1; HEPATOTOXICITY; METABONOMICS; BIOFLUIDS; STORAGE
subject category
Chemistry
authors
Graca, G; Duarte, IF; Goodfellow, BJ; Barros, AS; Carreira, IM; Couceiro, AB; Spraul, M; Gil, AM
our authors
Ana Maria Pissarra Coelho Gil
Associate Professor with Aggregation
António de Sousa Barros
Post-doc Fellowship
Brian James Goodfellow
Assistant Professor