This article presents background information related to methodology for estimating brain metabolite concentration from magnetic resonance spectroscopy (MRS) and magnetic resonance spectroscopic imaging measurements of living human brain tissue. It reviews progress related to this methodology, with emphasis placed on progress reported during the past 10 years. It is written for a target audience composed of radiologists and magnetic resonance imaging technologists. It describes in general terms the relationship between MRS signal amplitude and concentration. It then presents an overview of the many practical problems associated with deriving concentration solely from absolute measured signal amplitudes and demonstrates how a various signal calibration approaches can be successfully used. The concept of integrated signal amplitude is presented with examples that are helpful for qualitative reading of MRS data as well as for understanding the methodology used for quantitative measurements. The problems associated with the accurate measurement of individual signal amplitudes in brain spectra having overlapping signals from other metabolites and overlapping nuisance signals from water and lipid are presented. Current approaches to obtaining accurate amplitude estimates with least-squares fitting software are summarized.