Extraction methods of biologically active compounds of essential oil from marjoram (origanum majorana) and their applications in pharmaceutical formulation

Abstract

Objective: The main objective of the present work is to perform the phytochemical study investigating some biological and therapeutic properties such as antioxidant, antimicrobial, anticancer effects of Origanum majorana’s essential oil (OMEO), and its toxicity with wound healing properties in animals. Materials and Methods: An ethnobotanical study is conducted on Origanum majorana in the local population of Boumerdes. the essential oil was obtained using hydro-distillation technique with; later, it is subjected to chemical analysis using GC-MS. Assays of OMEO’s antioxidant activity by DPPH, and ABTS were carried out. The antibacterial activity is determined by disc diffusion and minimum inhibitory concentration techniques against different bacterial and fungal types. The anticancer activities are performed by MTT assays, Apoptosis assay, and migration assay, on PC-3 and SKBR3 cancer cell lines. In vivo toxicity is carried out on animal models, and the wound healing effects of a developed cream are studied using a cream formulation on induced wounds. Results: The ethnobotanical survey revealed that in 500 individuals questioned only 21% knew and used Origanum majroana. Regarding the essential oil’s hydrodistillation extraction yielded 3.33%. The analysis by GC-MS , revealed 29 compounds, which represent 98.08% of total volatile oil. The major compounds identified in OMEO were terpinen-4-ol (21.37%), ?-terpinene (15.78%), a-terpinene (10.43%), and trans-sabinene hydrate (9.27%). The assessement of the antioxidant activity showed different reactions to free radical scavenging activity with IC50 of 11.7mg/ml , and 265.2µg/ml, for DPPH , and ABTS, respectively, compared to BHT that exhibited an important activity with IC50 of 2.85µg/ml, and, 11.19µg/ml, respectively for DPPH and ABTS. Furthermore, The oil expresses a notable antimicrobial effect against most of the tested strains, the bacteria exhibit inhibition zones ranging from 10 to 31 mm and IC50 values from ?15.6 to 1051.9 ?g/ml for bacteria. Regarding the antifungal activity, the minimal inhibitory concentration (MIC) of the oil shows good results for Malassezia furfur, Candida albicans, Candida glabrata, Trichosporon sp., and Aspergillus niger, ranging from 0.125 to 0.250 ?l/ml. In the anticancer activity, the cytotoxicity evaluated by MTT on prostate cancer (PC-3), breast cancer (SKBR3), and normal retinal pigment epithelium (ARPE19) cell lines. The results show a selective cytotoxicity effect, by decreasing cell viability of PC-3 cancer cells with half Inhibitory Concentration (IC50) of 608.57µg/ml and 672.5µg/ml after 48h and 72h respectively. Regarding SKBR3 cancer cells the IC50 was 991.5µg/ml. OMEO exhibites no significant cytotoxicity against normal (ARPE19) cells. Furthermore, we conducted a cell apoptosis assay using Hochest 33342 dye to explore the potential mechanism pathway of OMEO. The findings verifie that OMEO could trigger apoptosis in PC3 and SBKR3 cancer cells. The ability of OMEO to inhibit cell

migration

assessed via wound healing assay reveals a significant decrease in cell migration. Acute toxicity study in vivo proves a moderate toxicity of 2500g/kg of the essential oil for therapeutic doses. Moreover, the test of wound healing reveals that 1% cream shows the most potent effect in tissue regeneration among the treated groups. Conclusion: The present study opens a window on the promising therapeutic potentials of Origanum majorana essential oil as a natural antioxidant, antimicrobial, and anticancer agent. Its pharmaceutical value is increasing by selective cytotoxicity and wound healing potential. However, additional studies need to be carried out to understand the mechanisms of action and explore possible clinical applications