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Previous work has demonstrated that deficiencies in volume selection sequences used in magnetic resonance spectroscopy may compromise the quality of the spectra obtained. In this paper, further studies on the ISIS and PRESS sequences are presented. Under conditions of partial saturation, ISIS can exhibit serious contamination with extraneous signal, particularly when a small volume of interest (VOI) is selected. ISIS protocols should therefore use VOIs that are large relative to the target volume, and repetition times that are as long as practicable. In PRESS, contamination is found to be minimised by using a VOI that is small relative to the target volume, and to be independent of repetition time. PRESS performance is also independent of echo time, except when very short echo times are used. These results are consistent with previously published work on ISIS and PRESS, and it is now possible to establish generic features of these sequences and to understand the implications for quantitative spectroscopy. T(1)-weighting of contamination in ISIS can compromise both relative and absolute quantification techniques in several respects. Contamination in PRESS is largely independent of relaxation times and would be easier to model and correct for in the context of quantitative spectroscopy.

We have performed a multicentre trial to assess the performance of three techniques for absolute quantification of cerebral metabolites using in vivo proton nuclear magnetic resonance (NMR). The techniques included were 1) an internal water standard method, 2) an external standard method based on phantom replacement, and 3) a more sophisticated method incorporating elements of both the internal and external standard approaches, together with compartmental analysis of brain water. Only the internal water standard technique could be readily implemented at all participating sites and gave acceptable precision and interlaboratory reproducibility. This method was insensitive to many of the experimental factors affecting the performance of the alternative techniques, including effects related to loading, standing waves and B1 inhomogeneities; and practical issues of phantom positioning, user expertise and examination duration. However, the internal water standard method assumes a value for the concentration of NMR-visible water within the spectroscopic volume of interest. In general, it is necessary to modify this assumed concentration on the basis of the grey matter, white matter and cerebrospinal fluid (CSF) content of the volume, and the NMR-visible water content of the grey and white matter fractions. Combining data from 11 sites, the concentrations of the principal NMR-visible metabolites in the brains of healthy subjects (age range 20-35 years) determined using the internal water standard method were (mean+/-SD): [NAA]=10.0+/-3.4 mM (n=53), [tCho]=1.9+/-1.0 mM (n=51), [Cr + PCr]=6.5+/-3.7 mM (n=51). Evidence of system instability and other sources of error at some participating sites reinforces the need for rigorous quality assurance in quantitative spectroscopy.