Mentha longifolia is a well-known species of medicinal plant for its ethnopharmacological, pharmacological importance, industrial uses etc. This research paper presents a comprehensive study on a new species Mentha longifolia subsp. himalaiensis essential oil, focusing on its GC-MS analysis, nematicidal and antioxidant activities, as well as in-silico PASS prediction. The essential oil was extracted from Mentha longifolia subsp. himalaiensis leaves using steam distillation and subjected to gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) analysis to identify its chemical composition. The in vitro nematicidal activities were accessed via nematode mortality assay and nematode egg hatching assay against Meloidogyne incognita. The essential oil was also tested for its antioxidant activities via DPPH (2,2-diphenyl-2-picrylhydrazyl) radical scavenging activity, H2O2 (Hydrogen peroxide) scavenging activity and Metal chelating activity. The essential oil was also tested for antimicrobial potential against 5 strains of bacteria and two strains of fungus. Furthermore, the major chemical constituents of the essential oils were subjected to in-silico PASS prediction for their biological activities. The GC and GC-MS analysis revealed that piperitenone oxide (31.13 %) and cis-piperitone oxide (51.61 %) were the major components of the essential oil. The essential oil exhibited significant nematicidal activity against M. incognita and notable antioxidant activity. The essential oils also showed significant antibacterial and antifungal activity against targeted microbial strains. In-silico PASS prediction further supported the essential oil's bioactive properties and its potential for diverse applications. These results highlight the potential of Mentha longifolia subsp. himalaiensis essential oil as a natural nematicide, antimicrobial and antioxidant agent.