Fungicides play an important role in controlling fungal diseases. Therefore, this study was conducted with the aim of evaluating the toxicity of fungicides on tomato spot disease pathogens under laboratory conditions. Nine chemical fungicides: Azoxystrobin + Difenoconazole, Copper Oxychloride, Chlorothalonil + Metalaxyl-M, Mancozeb, Dimethomorph + Pyraclostrobin, Tebuconazole, Azoxystrobin + Propiconazole, Chlorothalonil, and Metalaxyl-M were compared to one bio-fungicide Jojoba Oil on Alternaria alternata, Curvularia pseudobrachyspora, and Curvularia hawaiiensis using the poisoned food technique. Among the tested fungicides, tebuconazole demonstrated the highest potency in vitro inhibition (%) on C. hawaiiensis with an IC50 value of 0.8 ppm, while (Azoxystrobin + Difenoconazole) revealed the highest inhibition (%) on the growth of A. alternata and C. pseudobrachyspora with IC50 values of 0.83 and 1.88 ppm, respectively. The least inhibition was seen when copper oxychloride was used, with IC50 values of 4152.61, 3585.18, and 4755.95 for A. alternata, C. pseudobrachyspora, and C. hawaiiensis, respectively. A significant number of the tested fungicides are illustrated to be significantly effective in inhibiting the microorganisms that cause tomato spot disease, according to the overall results.