Localized magnetic resonance spectroscopy (LMRS) promises a powerful noninvasive means to determine myocardial triacylglycerol (TAG) in a clinical setting. Here, the linearity, specificity, robustness, precision, and accuracy, of an ex vivo mouse-heart LMRS TAG assay are assessed by quantifying the spatial, spectral, and relaxation induced uncertainties. The protocol, which is based on localization by adiabatic selective refocusing (LASER) using frequency offset corrected inversion pulses (FOCI), alternating gradient polarity, and simple post-processing, is shown to have good characteristics. The presented protocol has a benchmark, phantom-based, accuracy of 3%, and when applied to ex vivo mouse-hearts the accuracy is 6%, making the LMRS assay comparable to the typical destructive bioanalytical assay.