Thyroid benign (TBN) and malignant (TMN) nodules are a common thyroid lesion. The differentiation of TMN often remains a clinical challenge and further improvements of TMN diagnostic accuracy are warranted. The aim of present study was to evaluate possibilities of using differences in chemical elements (ChEs) contents in nodular tissue for diagnosis of thyroid malignancy. Contents of eight ChEs including bromine (Br), calcium (Ca), chlorine (Cl), iodine (I), potassium (K), magnesium (Mg), manganese (Mn), and sodium (Na) were prospectively evaluated in nodular tissue of thyroids with TBN (79 patients) and to TMN (41 patients). Measurements were performed using instrumental neutron activation analysis. It was observed that in TMN tissue the mass fractions of Br and I are approximately 2.9 and 14 times, respectively, lower, while the mass fraction of Ca, K, and Mg are 94%, 56%, and 36%, respectively, higher than in TBN tissue. Contents of Cl, Mn, and Na found in the TBN and TMN groups of nodular tissue samples were similar. Finally, it was proposed to use the I mass fraction and I/Ca, I/K, and I/Mg mass fraction ratios in a needle-biopsy of thyroid nodules as a potential tool to diagnose thyroid malignancy. Further studies on larger number of samples are required to confirm our findings and proposals.