PHYTOCHEMICAL COMPOSITION, ANTIMICROBIAL AND CYTOTOXIC ACTIVITIES OF CLINOPODIUM INSULARE ESSENTIAL OIL FROM PALESTINE
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Date
2023-02-05
Authors
Ziyad Bader Salman
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Abstract
ABSTRACT
Clinopodium insulare (Calamintha incana) is one of the medicinal aromatic plants that is dominant in the eastern Mediterranean regions, including Palestine, and has a distinctively pleasant mint-like smell. Traditionally, C. incana leaves are used as a spice and herbal tea. The current work's goal is to examine the potential pharmacological characteristics of chemical constituents.
The essential oil of the plant was extracted using the hydrodistillation (Clevenger method) technique. The use of gas chromatography–mass spectrometry (GC–MS), the chemical components of the plant's essential oil were determined. The essential oil's antioxidant capacity was evaluated by inhibiting 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical. The 3,5-dinitrosalicylic (DNSA) assay was used to assess anti-amylase activity. Anti-lipase activity was assessed using the p-nitrophynel butyrate method.
Antimicrobial activity was assessed using the broth microdilution method for antibacterial testing and the poisoned food technique for fungal testing. Six bacterial strains were used: four Gram-negative: Escherichia coli, Pseudomonas aeruginosa, Proteus vulgaris, and Klebsiella pneumonia; and one yeast: Candida albicans. Two were gram-positive: MRSA (Methicillin-Resistant Staphylococcus aureus); in addition, the oil was tested against three dermatophytes: Microsporum canis, Trichophyton rubrum, and Trichophyton mentagrophyte. Anti-cancer activity was evaluated by using the MTS assay on HeLa cells.
Plant essential oil yield was 0.213 percent (w/w). GC-MS analysis detected the presence of sixty compounds, which accounted for 98% of the total oil composition. The major compound was piperitone oxide, representing 41.178%. C. incana showed anti-oxidant activity with an IC50 of 390 μg/mL.. Essential oil has a moderate amylase inhibitory effect with an IC50 value of 120 μg/ml and a moderate anti-lipase inhibitory effect with an IC50 value less than 800 μg/ml. The sample showed broad antimicrobial activity, potent against Gram-negative bacteria with MICs ranging between 0.4883 ug/ml and 62.5 ug/ml , between 26.041 ug/ml and 31.25 ug/ml for Gram-positive bacteria, and yeast with a MIC of 0.2441 ug/ml. The antifungal activity against Microsporum canis, Trichophyton rubrum, and Trichophyton mentagrophyte was observed, with MICs ranging from 0.15 μl/ml against Trichophyton mentagrophyte to 0.37 μl/ml against Trichophyton rubrum. The anticancer activity of the oil was tested against HeLa cells and showed promising results with an IC50 dose of 50 μg/mL.
The plant's essential oil contained varying percentages of various phytochemicals, which provided various potential biological activities such as antioxidant, anti-lipase, anti-amylase, anti-cancer, and antimicrobial effects.
Keywords: Essential oil; Antibacterial activity; Antifungal activity; Clinopodium insulare.