Quantitative analysis of cardiac dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) perfusion datasets is dependent on the drawing (manually or automatically) of myocardial contours. The required accuracy of these contours for myocardial blood flow (MBF) estimation is not well understood. This study investigates the relationship between myocardial contour errors and MBF errors. Myocardial contours were manually drawn on DCE-MRI perfusion datasets of healthy volunteers imaged in systole. Systematic and random contour errors were simulated using spline curves and the resulting errors in MBF were calculated. The degree of contour error was also evaluated by two recognized segmentation metrics. We derived contour error tolerances in terms of the maximum deviation (MD) a contour could deviate radially from the 'true' contour expressed as a fraction of each volunteer's mean myocardial width (MW). Significant MBF errors were avoided by setting tolerances of MD ≤ 0.4 MW, when considering the whole myocardium, MD ≤ 0.3 MW, when considering six radial segments, and MD ≤ 0.2 MW for further subdivision into endo- and epicardial regions, with the exception of the anteroseptal region, which required greater accuracy. None of the considered segmentation metrics correlated with MBF error; thus, both segmentation metrics and MBF errors should be used to evaluate contouring algorithms.