The three chilli (Capsicum frutescens L.) cultivars viz. chilli DARL 210, Chilli Long Black and Sartoria were grown in middle hill climatic conditions of western Himalayas at an altitude of 5,500 feet above the sea level. The variation in total capsaicinoids (Capsaicin and dihydrocapsaicin) was studied from budding to full maturity stage of fruits. The fruits were harvested at every fifteen days interval from flower budding (stage zero) to full maturity stage (stage seven). The total capsaicinoids were evaluated with the help of HPLC. It can be concluded from the study that, to achieve maximum pungency (total capsaicinoids) in these cultivars, the chilli cv. DARL 210 should be harvested at the stage of full maturity (after 105 days). Whereas, Chilli long black should be harvested after 75 days and cultivar Sartoria in between 60 to 90 days from flower budding stage. As far as pungency among these cultivars is concerned DARL 210 was the most pungent followed by Chilli Long black, while Sartoria was found least pungent cultivar.