Equine Odontoclastic Tooth Resorption and Hypercementosis (EOTRH) is a common, painful and poorly understood disease. Enamel, dentin and cementum accumulate both essential and toxic trace elements during mineralization. Characterization of the spatial accumulation pattern of trace elements may provide insight into the role that toxic elements play and inform biological processes affecting these hard dental tissues for future research. Laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) was used to map the distribution of multiple trace elements and heavy metals across equine healthy and diseased (hypercementosis-affected) hard dental tissues among four teeth extracted from horses with EOTRH. Results showed banding patterns of some trace elements (lead, strontium, barium), reflecting the temporal component of accumulation of trace elements during dentin mineralization. Essential elements zinc and magnesium did not show banding patterns. Comparison to the unaffected cementum and dentin adjacent to the hypercementosis region showed that there is an underlying incremental pattern in the uptake of some metals with spatial irregularities. This supports a possible metabolic change involved in hypercementosis lesion development. This represents the first use of LA-ICP-MS to study the microspatial distribution of trace elements in equine teeth, establishing a baseline for elemental distribution in normal and EOTRH impacted dental hard tissue.