Quantification of myocardial perfusion by contrast-enhanced cardiovascular magnetic resonance imaging (CMR) aims for an observer independent and reproducible risk assessment of cardiovascular disease. Currently, the data used for the pixel-wise analysis of cardiac perfusion are either filtered prior to a fitting procedure, which inherently reduces the spatial resolution of data; or all pixels are considered without any regularization or prior filtering, which yields an unstable fit in the presence of low signal-to-noise ratio. Here, we propose a new pixel-wise analysis based on spatial Tikhonov regularization which exploits the spatial smoothness of the data and ensures accurate quantification even for images with low signal-to-noise ratio. The regularization parameter is determined automatically by an L-curve criterion. We study the performance of our method on a numerical phantom and demonstrate that the method reduces significantly the root-mean square error in the perfusion estimate compared to a non-regularized fit. In patient data our method allows us to recover the myocardial perfusion and to distinguish between healthy and ischemic regions.