A practical, rapid and inexpensive method for muscle glucose determination is proposed. The method is based on acid digestion of the sample, followed by neutralization of the solution and the subsequent glucose determination using a domestic glucometer for human blood glucose level control. In a pre-experimental phase, the accuracy of this methodology and the titration method was determined, comparing results to pre-established glucose concentrations. There were significant differences between the glucose reading given by the glucometer and the pre-established glucose concentrations and the results given by the titration method. Nevertheless, when discrepancies (bias) were identified, differences resulted non-significant (P = 0.85). Subsequently, muscle glucose concentration (n = 24) was validated in the experimental phase and significant differences were shown between glucose concentration values given by the glucometer and the titration method, the latter considered as a standard method or as a reference (P < 0.0001). Once the sources of discrepancies were identified and removed, the differences observed between the values given by the two methods were not statistically significant (P = 0.99). It was concluded that the glucometer is a potential method for muscle glucose determination, but must be improved.