Spectra were obtained using the Bruker 1-D NOESY gradient water suppression pulse sequence ‘noesygppr1d’ with 10 ms mixing time. Each sample was run for 512 scans to a total acquisition size of 128 k, a spectral window of 20.5 ppm, a transmitter offset of 4.7 ppm, and a recycle delay of 4 s. All measurements were recorded using a Bruker triple resonance TBO-Z probe at room temperature. The above acquisition parameters correspond to an approximate data collection time of 1 h per sample. The Bruker automation program “pulsecal” was used on each sample before data acquisition to guarantee that the 90-degree pulse was calibrated correctly, ensuring quantitative and comparable data across samples (Paxman et al., 2018 (link)). The spectra were zero filled to 256 k, automatically phased, baseline corrected, and line-broadened by 0.3 Hz (Kiss et al., 2016 (link)). Spectra were then exported to MATLAB (MathWorks, Natick, MA) as ASCII files where they underwent Dynamic Adaptive Binning (Anderson et al., 2011 ), followed by manual inspection and correction. The dataset was then normalized to the total metabolome, excluding the region containing the water peak, and pareto scaled and log-transformed.
1 d noesy gradient water suppression pulse sequence
Manufactured by Bruker
The 1-D NOESY gradient water suppression pulse sequence is a nuclear magnetic resonance (NMR) technique used to acquire one-dimensional NMR spectra with efficient suppression of the water signal. The core function of this pulse sequence is to selectively suppress the intense water resonance while preserving the signals of interest in the NMR spectrum.
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3 protocols using 1 d noesy gradient water suppression pulse sequence
1
NMR Spectroscopic Analysis of Metabolomes
2
NMR Spectroscopy for Metabolomic Analysis
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Spectra were collected on a 700 MHz Bruker Avance III HD spectrometer (Bruker, Milton, ON, Canada). The Bruker 1-D NOESY gradient water suppression pulse sequence ‘noesygppr1d’ was used with 10 ms mixing time. Each sample was run for 512 scans to a total acquisition size of 128 k, a spectral window of 20.5 ppm, a transmitter offset of 4.7 ppm, and a recycle delay of 4 s. All measurements were recorded using a Bruker triple resonance TBO-Z probe. The Bruker automation program “pulsal” was used on each sample before data acquisition to guarantee that the 90-degree pulse was calibrated correctly, ensuring quantitative and comparable data across samples [26 (link)]. The spectra were zero filled to 256 k, automatically phased, baseline corrected, and line-broadened by 0.3 Hz [25 (link)]. Spectra were then exported to MATLAB (MathWorks, Natick, MA, USA) as ASCII files, and underwent dynamic adaptive binning [30 (link)], followed by manual inspection and correction. Spectral binning resulted in 439, 460, and 379 spectral bins for kidney, liver, and breast muscle, respectively. The dataset was then normalized to the total metabolome, excluding the region containing the water peak, and pareto scaled.
3
NMR Profiling of Concussion Biomarkers
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NMR spectra were collected on a 700-MHz Bruker Avance III HD spectrometer (Bruker, ON, Canada). The Bruker 1D NOESY gradient water suppression pulse sequence was used. Each sample was run for 128 scans, and the total acquisition size was 128 k. The spectra were zero filled to 256 k, automatically phased, baseline corrected, and line-broadened to 0.3 Hz. The processed spectra were exported as ascii files to MATLAB version R2015b (The MathWorks, MA, USA) for statistical analysis. Spectra were first binned using dynamic adaptive binning [code available for download at (28 (link))] and then manually adjusted to optimize the number of variables and ensure accuracy. All spectra had the regions corresponding to water and urea removed before normalization to the total area of all spectral bins. The peaks corresponding to acetaminophen derivatives (29 (link)) were also removed to account for the fact that the athletes tended to take pain medication following an SRC. The data were then Pareto scaled to increase the influence of weak peaks and deemphasize the influence of larger peaks. All the peaks in each spectrum were referenced to TSP (0.00δ).
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