| Summary: | <i>Cymbopogon</i> species essential oil (EO) carries significant importance in pharmaceuticals, aromatherapy, food, etc. The chemical compositions of <i>Cymbopogon</i> spp. <i>Viz. Cymbopogon winterianus</i> (citronella) <i>Cymbopogon citratus</i> (lemongrass), and <i>Cymbopogon martini</i> (palmarosa) were analyzed by gas chromatography–mass spectrometry (GC-MS), enantiomeric distribution by chiral GC-MS, and antimicrobial activities of some selected pure major compound and root and leaves EOs of citronella. The EO of leaves of <i>Cymbopogon</i> spp. showed comparatively higher yield than roots or other parts. Contrary to citral (neral and geranial) being a predominant compound of <i>Cymbopogon</i> spp., α-elemol (53.1%), α-elemol (29.5%), geraniol (37.1%), and citral (90.4%) were detected as major compounds of the root, root hair with stalk, leaf, and root stalk with shoot of citronella EO, respectively. Palmarosa leaves’ EO contains neral (36.1%) and geranial (53.1) as the major compounds. In the roots of palmarosa EO, the prime components were α-elemol (31.5%), geranial (25.0%), and neral (16.6%). Similarly, lemongrass leaves’ EO contains geraniol (76.6%) and geranyl acetate (15.2%) as major compounds, while the root EO contains a higher amount of geraniol (87.9%) and lower amount of geranyl acetate (4.4%). This study reports for the first time chiral terpenoids from <i>Cymbopogon</i> spp. EOs. Chiral GC-MS gave specific enantiomeric distributions of nine, six, and five chiral terpenoids in the root, root stalk with a shoot, and leaves of citronella EOs, respectively. Likewise, four and three chiral terpenoids in the root and leaves of lemongrass oil followed by two chiral terpenoids in the leaves and root of palmarosa EOs each. Additionally, the root and leaves’ EOs of citronella exhibit noticeable activity on bacteria such as <i>Staphylococcus aureus</i>, <i>Staphylococcus epidermidis</i>, and <i>Streptococcus pyogenes</i> and fungus such as <i>Candida albicans</i>, <i>Microsporum canis</i>, and <i>Trichophyton mentagrophytes</i>. So, geranial-, neral-, geraniol-, and citronellal-rich EOs can be used as an alternative antimicrobial agent.
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