Serum Metabolite Profiling via NMR

Serum metabolites of samples collected from three time-points i.e., baseline (prior to drink consumption), 30 and 120min of OGTT were analyzed and profiled by ¹H NMR spectroscopy (Bruker Advance 600, Canada) as previously described by our laboratory [16 (link)]. Briefly, serum samples (350 μL) were first filtered through pre-washed 10-kDa ultra centrifugal filters, and the filtrate was transferred to phosphate buffer containing NaN₃ and dimethyl silapentane sulfonate (DSS). Samples were brought to a final volume of 450 μL with D₂O, so that the concentration of DSS in the sample remained at 0.5M, before the analysis. Resultant data was individually processed and profiled using Chenomx NMR Suite 7.5 (Chenomx, Canada).

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Mayengbam S., Virtanen H., Hittel D.S., Elliott C., Reimer R.A., Vogel H.J, & Shearer J. (2019). Metabolic consequences of discretionary fortified beverage consumption containing excessive vitamin B levels in adolescents. PLoS ONE, 14(1), e0209913.

Publication 2019

Advance 600 Chenomx NMR Suite 7.5

Buffer Nan3 Nmr spectroscopy Ogtt Phosphate Serum Sulfonate

Corresponding Organization : University of Calgary

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Variable analysis

independent variables

Time-points (baseline, 30min, 120min) of OGTT

dependent variables

Serum metabolites profiled by 1H NMR spectroscopy

control variables

Serum sample volume (350 μL)
Phosphate buffer containing NaN3 and dimethyl silapentane sulfonate (DSS)
Final sample volume (450 μL) with 0.5M DSS concentration

controls

Positive control: Not mentioned
Negative control: Not mentioned

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