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Not AvailableCapsicum species are not only cultivated as vegetable and condiment crop but are also incorporated into a number of medicinal preparations in the ancient literature around the world. ‘Naga chilli’ or ‘Bhoot Jolokia’ (Capsicum chinense Jacq.) is a chilli variety indigenous to the northeast region of India and has been recognized as one of the hottest chilli in the world. It has also been used conventionally in treating various human ailments since time immemorial by the indigenous people of the northeast India. Capsaicin and dihydrocapsaicin are the two major members of the so-called capsaicinoid family, which includes other minor analogues, and usually account for at least 90 % of the pungency trait in chilli fruits. In the present study, determination of Capsaicinoid content of Bhoot Jolokia (Capsicum chinense Jacq.) under different sowing time and spacing was done. In the Scoville Organoleptic test, the highest Scoville Heat Unit (SHU) value was found in September 15 sowing fruits and the lowest in the February 15 sowing fruits. Fruit produced from spacing 105 cm × 105 cm showed highest Scoville Heat Unit. In High Performance Liquid Chromatography analysis, highest amount of capsaicinoid was also found in September 15 sowing while fruits produced from spacing 105 cm × 105 cm showed highest amount of capsaicinoid. Identification of Nordihydrocapsaicin, Capsaicin and Dihydrocapsaicin was done by comparing the retention time of sample with those of standard.Not Availabl