In vivo magnetic resonance spectroscopy currently lacks data-driven quality control standards, the development of which can be informed by characterizing the effects of spectral quality on metabolite quantification. We assess the influences of spectral line width and signal-to-noise ratio on both simulated and experimentally derived in vivo resonances from twenty-one metabolites measured by short-echo-time STEAM and J-difference editing for glutathione and GABA at 7 Tesla. We show that spectral quality exerts distinct effects on apparent concentrations of different metabolites, underlining the importance of explicit quality analysis in studies employing in vivo magnetic resonance spectroscopy, especially cross-sectional investigations of physiologically distinct groups.
The in vivo magnetic resonance spectroscopy (1H-MRS) field needs data-driven quality control standards.1 Such standards can be informed by quantifying the effects of spectral quality, including full width at half maximum (FWHM; frequency width of a resonance at the ordinate point halfway between baseline and apex) and signal-to-noise ratio (SNR; a prominent singlet amplitude normalized by noise intensity standard deviation) on metabolite quantification.
Understanding the numerical influences of spectral quality on metabolite quantification informs both quality control and experimental integrity. Brain water or metabolite spectral FWHM has demonstrated increases in multiple sclerosis,2 Wilson’s disease,3 and normal maturation.4 Additionally, any cohort who might compromise magnetic field homogeneity by increased motion between shim optimization and signal acquisition, like children, may also exhibit broader lines. It is important to determine how this may influence metabolite quantification by 1H-MRS. Some previous work has addressed this question with simulation experiments,5-7 but these findings are not generalizable to higher field strengths or to metabolite signals isolated by spectral editing.
Glutathione (GSH) is an antioxidant synthesized in neurons and glia,8 while γ-amino butyric acid (GABA) is an inhibitory neurotransmitter and potential immunomodulator9 and glutamate the central nervous system's predominating excitatory neurotransmitter. All three compounds play key mechanistic roles in neurological health and disease. Reduced peak amplitude and complex line shapes due to J-coupling-induced signal splitting, plus spectral overlap with nearby resonances, complicate the spectroscopic quantification of these molecules, predicting its strong influence by spectral quality.
To facilitate data-driven quality assessment for 1H-MRS experimental planning and interpretation, we applied linear combination modeling and quantification error analysis to 25,250 simulated and measured MEGA-sLASER and STEAM spectra based on previously published sequences at 7 Tesla10 and calculated the effects of spectral quality measures FWHM and SNR on the quantification accuracy of these three metabolites, plus common concentration references total creatine and N-acetyl aspartate (NAA).
We analyzed the accuracy of linear combination model fits to proton spectra of varying complexity to examine how FWHM and SNR affect apparent metabolite concentration as quantified by in vivo 1H-MRS at 7 T. Linear combination modeling routines exploited functionality from INSPECTOR;11 additional scripting was implemented in MATLAB (Mathworks, Natick, MA, USA). Quantification errors were calculated relative to apparent concentrations output by perfect model fits to noiseless spectra at each FWHM and signal amplitude combination (Figure 1A).
Condition 1 comprised fits at 5 FWHM, 10 starting signal amplitudes, and 100 Gaussian noise patterns, based on a simulated Lorentzian singlet at 3.01 ppm, or GSH or GABA difference resonances from MEGA-sLASER experiments (TE = 72 ms) with J-difference editing (glutathione 4.56 ppm; GABA 1.89 ppm)10 simulated using SpinWizard12 (Figure 1B). Actual concentration was kept constant across FWHM. Noise patterns were matched across signal amplitude and line width conditions for each resonance; noise amplitude remained constant within each experiment. Condition 2 consisted of fits at 5 FWHM and 5 starting signal amplitudes, also with 100 Gaussian noise patterns, based on GSH or GABA difference spectra plus co-edited resonances (NAA or NAA, glutamate, and glutamine, respectively), or full metabolite spectra (STEAM, TE = 10 ms)10 including twenty-one metabolites, in approximately physiological ratios (Figure 1C). Condition 3 repeated GSH J-difference spectral fits with an in vivo baseline averaged from fit residuals to similar acquisitions in the frontal cortex of healthy adults (N = 7). This enabled the calculation of quantification error under spectral conditions resembling those seen in vivo against a perfect gold standard without noise or baseline (Figure 1D).
Results
Fit error standard deviation decreased with SNR (Figures 2, 3B) and increased with FWHM (Figures 2, 3C) in all fits of singlets, GABA, and GSH and varied positively with metabolite amplitude Cramér-Rao Lower Bound (CRLB) (Figures 2, 3D-E). In vivo baseline conditions introduced systematic asymmetry into quantification errors expected from linear combination model fits plus linear baseline to GSH J-difference spectra (Figure 3B, C), and glutamate fit errors exhibited greater influence by spectral quality than those of total creatine or NAA (Figure 4). Underestimation of actual fit errors by reported CRLB varied with fit frequency range and baseline complexity (Figure 5).
Conclusions
SNR and FWHM predictably affect the standard deviation of quantification errors expected from linear combination model fitting of isolated and overlapping singlet, J-difference-edited, and unedited metabolite resonances. Complex baseline shapes may introduce systematic quantification errors that could influence conclusions based on comparisons of groups exhibiting disparate spectral quality. These results motivate further work on the effects of fitting algorithms, particularly baseline modeling routines, on conditions exhibiting such systematic errors here. All told, the relationship between spectral quality and fit error differs by metabolite, arguing against the use of one-size-fits-all quality control thresholds for in vivo 1H-MRS experiments.
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