Quantification of magnetic resonance spectroscopy data using a combined reference: Application in typically developing infants

NMR Biomed. 2021 Jul;34(7):e4520. doi: 10.1002/nbm.4520. Epub 2021 Apr 28.


Quantification of proton magnetic resonance spectroscopy (1 H-MRS) data is commonly performed by referencing the ratio of the signal from one metabolite, or metabolite group, to that of another, or to the water signal. Both approaches have drawbacks: ratios of two metabolites can be difficult to interpret because study effects may be driven by either metabolite, and water-referenced data must be corrected for partial volume and relaxation effects in the water signal. Here, we introduce combined reference (CRef) analysis, which compensates for both limitations. In this approach, metabolites are referenced to the combined signal of several reference metabolites or metabolite groups. The approach does not require the corrections necessary for water scaling and produces results that are less sensitive to the variation of any single reference signal, thereby aiding the interpretation of results. We demonstrate CRef analysis using 202 1 H-MRS acquisitions from the brains of 140 infants, scanned at approximately 1 and 3 months of age. We show that the combined signal of seven reference metabolites or metabolite groups is highly correlated with the water signal, corrected for partial volume and relaxation effects associated with cerebral spinal fluid. We also show that the combined reference signal is equally or more uniform across subjects than the reference signals from single metabolites or metabolite groups. We use CRef analysis to quantify metabolite concentration changes during the first several months of life in typically developing infants.

Keywords: combined reference, GABA, infant development, magnetic resonance spectroscopy, quantification, water scaling.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Corpus Callosum / diagnostic imaging
  • Data Analysis*
  • Female
  • Gray Matter / diagnostic imaging
  • Humans
  • Infant
  • Magnetic Resonance Spectroscopy*
  • Male
  • Metabolome
  • Reference Standards
  • Signal Processing, Computer-Assisted
  • Water
  • White Matter / diagnostic imaging


  • Water