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The composition of L. cubeba oil has been reviewed in a few literatures.12,13 Literature search revealed that the chemical composition of litsea oil varies with different geographic areas and thereby the synergistic or antagonistic effects cause different biological activities.7,14-17 Several authors have reported that citral (α-citral; geranial and β-citral; neral) is the major constituent in the oil.7,11,18-21 In Indian litsea oil, Gogoi et al. have reported that citral (geranial 51.9% and neral 40.7%) was the major constituent.18 On the other hand, citronellal (44.8-77.2%) and citronellol (10.9-14.0%) were the major constituents but not citral (geranial 0.20.4%) in litsea oil from Assam, India.22 Litsea oil of Chinese variety contained nerol, β-caryophyllene, 2,6-dimethyl-5-heptenal and 6-methyl-5-hepten-2ol as major constituents.22 On the other hand, the major constituents of Tibetian, Chinese litsea oil were limonol (44.2%), linalool (8.8%), eucalyptol (5.4%), elemicin (3.9%), methyl euginol (3.8%), etc.8 In litsea oil from Jiangxi province of China, Si et al.15 have reported that citral (80.6-82.9%) and limonene (4.1-5.3%) were the major constituents, while the composition was varied with citral (47.9%) and limonene (26.2%) in a report of Wang et al.23 The ethanolic extract of L. cubeba fruits from Zhejiang Province, China was analysed by Hu et al., and thirty compounds were identified with citral (59.3%) and limonene (8.5%) as the predominant compounds.14 Therefore, the composition of litsea oil remarkably varied within India and China. Since Nepal lies between these two countries, we decided to compare the variation in the oil composition from our own product to the global distribution and its affects in the bioactivities. Herein, we report the identification and quantification of volatiles of litsea oil by GCMS, phytochemical screening of the fruit extracts, and in vitro evaluation of antibacterial activity and antioxidant capacity of the oil and extracts. ABSTRACT Background: Biological activities of the plant materials may vary with different geographic origin. Litsea cubeba fruits are consumed in Nepal as a spice ingredient and are considered to be possessed antibacterial property. Objectives: Aims of this work are: to compare phytoconstituents present in litsea oil by GC-MS analysis, and to evaluate antibacterial and antioxidant activities of the volatile oil and the fruit extracts. Materials and Methods: Hydrodistillation of L. cubeba fruits was performed to obtain litsea oil followed by extractions with boiling methanol. Fractionations of the extract provided hexane, dichloromethane, ethyl acetate and aqueous extracts. The oil and extracts were analysed for the phytochemicals present. They were evaluated by using antibacterial susceptibility, brine-shrimp lethality and antioxidant capacity assays. Results: All together 49 compounds (accounting 93.66%) were identified in the litsea oil by GC-MS. Monoterpenes being dominant components, the oil constituted 15.96% of citral (geranial and neral). Other major constituents were capric acid (12.44%), β-caryophyllene oxide (7.69%), linalool (5.96%), eucalyptol (5.13%) and cisβ-terpineol (4.22%). Litsea oil, hexane extract and dichloromethane extract displayed very effective antibacterial property. Ethyl acetate extract (LC50 = 21.52 μg mL ‒1) and litsea oil (LC50 = 31.62 μg mL‒1) were found highly cytotoxic against brine-shrimp nauplii. Ethyl acetate (IC50 = 124.57 μg mL‒1) and dichloromethane (IC50 = 271.08 μg mL ‒1) extracts displayed a modest DPPH free radical scavenging activity. Conclusion: Phytoconstituents present in the L. cubeba fruits from Nepal were analysed. Litsea oil and the extracts have displayed high antibacterial and potentially anticarcinogenic activities.

A High Antibacterial Efficacy of Fruits of Litsea cubeba (Lour.) Pers from Nepal. GC-MS and Antioxidative Capacity Analyses