Osteoporosis is a systemic skeletal disease characterized by a disorder between bone reabsorption and formation with increase in bone fragility. The aim of this study was to evaluate the bone mass loss using solid-state techniques and biochemical assays to characterized bone samples of ovariectomized rats (OVX) and control group (CTR). The in vivo assays were carried out using biochemical analyses and bone mineral density (BMD) measurements. After the animals were euthanized, the femurs were examined using simultaneous differential thermal analysis (DTA), thermogravimetry/derivative thermogravimetry (TG/DTG) and atomic absorption spectrometric. The microstructure was examined using scanning electron microscopy (SEM). OVX had statistically higher averages (p < 0.05) for alkaline phosphatase activity in the serum. Therefore, our results demonstrated that the comparison of the OVX with the CTR showed no evidence of significant differences in the serum calcium and phosphorus levels (p > 0.05); however, OVX showed a significant weight gain (p < 0.0001) with a body mass increase of 30.23 ± 6.45 %. BMD results presented significant differences (p = 0.01985) between CTR and OVX. The total mass loss (TG) was 39.12 ± 0.45 % in OVX, while in CTR groups, it was the 32.45 ± 1.01 % (p = 0.0038). The results of the calcium amount in the bones of OVX (199.86 ± 8.08) were lower than the CTR (229.62 ± 10.02). SEM micrographs showed that OVX presented bone porosity, while CTR showed denser trabecular structure; this result corroborated with bone mass loss in OVX. The ovariectomy-induced bone loss in the rodent model can be evaluated using physical–chemical techniques based on thermal analysis, atomic absorption spectrometry and scanning electronic microscopy.