Waste Citrus limon Leaves as Source of Essential Oil Rich in Limonene and Citral: Chemical Characterization, Antimicrobial and Antioxidant Properties, and Effects on Cancer Cell Viability
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Published:2023-06-08
Issue:6
Volume:12
Page:1238
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ISSN:2076-3921
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Container-title:Antioxidants
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language:en
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Short-container-title:Antioxidants
Author:
Petretto Giacomo Luigi1ORCID, Vacca Giuseppe1, Addis Roberta1, Pintore Giorgio1, Nieddu Mariella2ORCID, Piras Franca2, Sogos Valeria2ORCID, Fancello Francesco3ORCID, Zara Severino3ORCID, Rosa Antonella2ORCID
Affiliation:
1. Department of Medicine, Surgery and Pharmacy, University of Sassari, Viale San Pietro, 07100 Sassari, Italy 2. Department of Biomedical Sciences, University of Cagliari, Cittadella Universitaria, 09042 Monserrato, CA, Italy 3. Department of Agriculture, University of Sassari, Viale Italia, 07100 Sassari, Italy
Abstract
This study investigated chemical composition, cytotoxicity in normal and cancer cells, and antimicrobial and antioxidant activity of the essential oil (EO) isolated by hydrodistillation from the discarded leaves of lemon (Citrus limon) plants cultivated in Sardinia (Italy). The volatile chemical composition of lemon leaf EO (LLEO) was analyzed with gas chromatography-mass spectrometry combined with flame ionization detection (GC/MS and GC/FID). The most abundant component of LLEO was limonene (260.7 mg/mL), followed by geranial (102.6 mg/mL) and neral (88.3 mg/mL). The antimicrobial activity of LLEO was tested using eight bacterial strains and two types of yeasts by a microdilution broth test. Candida albicans showed the greatest susceptibility (MIC = 0.625 μL/mL) and Listeria monocytogenes and Staphylococcus aureus were inhibited at low LLEO concentration (MIC values from 2.5 to 5 μL/mL). The C. limon leaf EO displayed radical scavenging ability (IC50 value of 10.24 mg/mL) in the 2,2-diphenyl-1-picryl-hydrazylhydrate (DPPH) assay. Furthermore, the LLEO impact on cell viability was explored by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in cancer HeLa cells, A375 melanoma cell line, normal fibroblasts (3T3 cells), and keratinocytes (HaCaT cells). LLEO, at 24 h of incubation, significantly reduced viability from 25 μM in Hela cells (33% reduction) and A375 cells (27%), greatly affecting cell morphology, whereas this effect was found from 50 μM on 3T3 fibroblasts and keratinocytes. LLEO’s pro-oxidant effect was also established in HeLa cells by 2′,7′-dichlorodihydrofluorescein diacetate assay.
Funder
Research Integrative Fund (FIR) of the University of Cagliari
Subject
Cell Biology,Clinical Biochemistry,Molecular Biology,Biochemistry,Physiology
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