Mentha × piperita L. is one of the highly demanded Mentha species reputed for commercial peppermint oil. In present research, the chemical characterization of the essential oil of a novel menthofuran rich variant of peppermint (Mentha × piperita L.) was done using gas chromatography coupled with mass spectrometry. Moreover, in order to explore the potential for its industrial use, the variability in essential oil yield and composition was analysed and compared at different crop maturity of first and second harvest. Altogether, twenty-eight constituents representing 96.11-98.56% of essential oil compositions were identified, exclusively represented by oxygenated monoterpenes (88.56-92.40%), viz. menthofuran (32.88-42.32%), pulegone (22.25-37.69%), menthol (9.83-21.01%), and menthone (2.47-8.13%). Significant variations were noticed in the herb and oil yield, and content of major constituents due to different harvesting maturity of the crop. The results obtained could be of economic importance for obtaining higher yield of quality essential oil (with significant content of menthofuran and pulegone) under sub-tropical agro climatic conditions of north India.