Hot peppers are known for medicinally important capsaicinoids. Bhut Jolokia, Capsicum chinense/Capsicum frutescens, is the hottest pepper in the world. However, its bioactivity and quantity of heat principles are not reported. We have now quantified capsaicin (C) and dihydrocapsaicin (DHC) in Bhut Jolokia and compared it with commonly consumed hot peppers, Jalapeno (Capsicum annuum) and Scotch Bonnet (Capsicum chinense). The concentration of C and DHC in Bhut Jolokia was 5.36%, which is about 338 and 18 times greater than in Scotch Bonnet and Jalapeno, respectively. We have also isolated capsaicin (C) and dihydrocapsaicin (DHC) in pure form and determined lipid peroxidation (LPO) and cyclooxygenase (COX-1 and -2) enzymes inhibitory concentrations. This is the first report of the quantification of C and DHC in Bhut Jolokia, comparison of capsaicinoids content in Bhut Jolokia with Jalapeno and Scotch Bonnet hot peppers and the COX and LPO inhibitory activities of C and DHC.