There is a rapidly growing market for topical use of virgin coconut oil (VCO). Studies of topical use in dogs are lacking. The objective of this study was to measure the release of lactate dehydrogenase (LDH, a plasma membrane disruption marker) and production of nitrite (Griess reaction, an oxidative stress marker) from a canine keratinocyte cell line after exposure to VCO as an initial toxicity screening to suggest future studies. Canine progenitor epidermal keratinocytes (CPEKs) were plated onto permeable transwell membranes and cultured with undiluted organic VCO or control media. Following a 24h incubation, an LDH assay and a Griess reaction were performed on the collected subnatants. Exposure of CPEKs to VCO significantly increased LDH release compared to controls, 62.29±16.32% versus 8.88±5.82% (P=0.0056) and there was no significant difference in production of nitrite compared to controls, 2.47±1.56μmol/L versus 1.42±0.95μmol/L (P=0.086). Based on this study VCO induced an increased disruption of plasma membrane integrity, as measured by LDH. However, VCO did not induce increased oxidative stress, as measured by nitrite production. Based on these preliminary data, further studies to assess the toxicity of VCO are needed.