Constant time gradient HSQC‐iDOSY: practical aspects

McLachlan, Andy S.; Richards, J. J.; Bilia, Anna Rita; Morris, Gareth A.

doi:10.1002/mrc.4277

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“…(vi) 1 H‐ 13 C HSQC experiments incorporating an MT module (MT‐HSQC), performed using a 2‐s CW B 1 = 1 μT saturating field placed at either 6.7 or 8.2 ppm, and four phase‐cycled averages; the output of these scans was subtracted from identical experiments involving the same timings but no saturation, in order to observe the moieties affected by the CW irradiation. (vii) 1 H‐ 13 C HSQC experiments incorporating diffusion weighting via a constant time, stimulated‐echo, bipolar gradient pulsed sequence, 46 using 2‐ms diffusion gradient pulses of 5 and 25 G/cm, separated by a diffusion time of 21.7 ms; in order to account for the loss of sensitivity because of the diffusion weighting, each experiment was performed in eight blocks of four averages, which alternated between high and low b‐valued experiments; these were then suitably combined, in order to highlight fast‐diffusing 1 H‐ 13 C pairs.…”

Section: Methodsmentioning

confidence: 99%

High‐field ex vivo and in vivo two‐dimensional nuclear magnetic resonance spectroscopy in murine brain: Resolving and exploring the molecular environment

2022

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The structural and chemical complexities within the brain pose a challenge that few noninvasive techniques can tackle with the dexterity of nuclear magnetic resonance (NMR) spectroscopy. Still, even with the advent of ultrahigh fields and of cryogenically cooled coils for in vivo research, the superposition of metabolic resonances arising from the brain remains a challenge. The present study explores the potential to tackle this milieu using a combination of two-dimensional (2D) NMR techniques, implemented on murine brains in vivo at 15.2 T and ex vivo at 14.1 T. While both experiments were affected by substantial inhomogeneous broadenings conveying distinct elongated lineshapes to the cross-peaks, the ability of increased fields to resolve off-diagonal resonances was clear. A comparison between the corresponding conventional and double quantum-filtered correlated spectroscopy traces enabled an improved assignment of in vivo resonances on the basis of more sensitive ex vivo 2D acquisitions, foremost on the basis of homonuclear cross-relaxation-driven correlations for peaks resonating downfield from water, and of heteronuclear correlations at natural abundance for the upfield protons. With the aid of such 2D correlations approximately 29 metabolites could be resolved and identified. This enhanced resolution was used to explore features related to the metabolites' diffusivities, their exposure to water, and their facility to undergo magnetization transfers to amide/amine/ hydroxyl resonances. Cross-peaks from main murine brain biomolecules, including choline, creatine, γ-aminobutyric acid, N-acetyl aspartate, glutamine, and glutamate, showed enhancements in several of these various features, opening interesting vistas about metabolite compartmentalization as viewed by these 2D NMR experiments.

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Section: Methodsmentioning

confidence: 99%