Mustard oil is popular edible oil in Bangladesh. It contains about more than fifty percent erucic acid which is toxic to human beings. This study evaluated the determination of biochemical properties (acid value, iodine value, saponification value and unsaponifiable matter %) of mustard oil, virgin coconut and sesame oil and toxicity determination of these oils through blood indices targeting of these oil by mixing with normal diet as a dietary supplementation. This study found acid value of mustard, sesame and virgin coconut oil was 0.92, 0.312, 0.52 respectively. Iodine value of mustard, sesame and virgin coconut oil were 110.2, 105.2, and 11 respectively. Saponification value of wild mustard, sesame and virgin coconut oil was 171.1, 187.2 and 240 respectively and unsaponifiable matter % of mustard, sesame and virgin coconut oil was 1.19, 0.31 and 0.92 respectively. Also in vivo experiment we divided wistar male rats were 4 groups such as normal diet 12 g/day/rat, mustard oil (MO) 0.6 g/day/rat, virgin coconut oil (VCO) 0.6g/day/rat and sesame oil (SO) 0.6 g/day/rat. In this study the vegetable oil causes the effect on average body weight gain, Food Efficiency Ratio (FER), and lipid profiles [Total cholesterol (Tch), HDL- Cholesterol LDL - Cholesterol, Triglyceride (TG)] and serum enzymes [SGOT (AST), SGPT (ALT)] of different groups of rat also been estimated. The study found that mustard oil significantly (p<0.05) increase lipid profile and enzymes SGOT, SGPT level in blood serum which is an indication of heart and liver disorder. The study found that coconut and sesame oil consumption in rat significantly (p<0.05) decrease of these parameters.