The aim of this study was to evaluate the mineral content of dentin prepared using three different desensitizing agents and the Nd:YAG laser. The occlusal third of the crowns of 30 molar teeth were cut with a slow-speed diamond saw sectioning machine under water cooling. Dentin slabs from the 30 teeth were randomly divided into five experimental groups, each comprising six slabs. The five groups were treated as follows: group A, no treatment; group B, treatment with oxalate-containing desensitizing agent (BisBlock); group C, treatment with resin-based desensitizing agent (Admira Protect); group D, treatment with glutaraldehyde-containing desensitizing agent (Systemp); and group E, irradiation with the Nd:YAG laser (DEKATM ) at 0.40 W. The levels of Mg, P, Ca, K, and Na in each slab were measured by inductively coupled plasma-atomic emission spectrometry (ICP-AES). Data were analyzed by one-way analysis of variance and the Tukey HSD test. The effect of desensitizing agents and laser irradiation on the dentin surface were evaluated using a scanning electron microscope. There were no significant differences between the groups (p > 0.05). Group E was showed the lowest Ca/P ratio. SEM showed that the resin-based agent occluded the dentinal tubules, the glutaraldehyde-containing agent increased the Ca/P ratio, and Nd:YAG laser irradiation decreased the Ca/P ratio. The mean percentages by weight of Ca, Mg, K, Na and P were not affected by Nd:YAG laser irradiation or any of the desensitizing agents.