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Trop J Nat Prod Res, December 2020; 4(12):1108-1115 ISSN 2616-0684 (Print) ISSN 2616-0692 (Electronic) Tropical Journal of Natural Product Research Available online at https://www.tjnpr.org Original Research Article Chemical Composition, Antioxidant Potential and Antimicrobial Properties of the Essential Oils of Haplophyllum tuberculatum (Forsskal) A. Juss from Morocco Abdelkrim Agour1*, Ibrahim Mssillou1, Hamza Saghrouchni2, Amina Bari2, Badiaa Lyoussi1, Elhoussine Derwich1,3 1Laboratory of Natural Substances, Pharmacology, Environment, Modeling, Health & Quality of Life, Department of Biology, Faculty of Sciences, Sidi Mohamed Ben Abdellah University, Fez, Morocco 2Laboratory of Biotechnology, Environment, Agri-food and Health, Department of Biology, Faculty of Sciences, Sidi Mohamed Ben Abdellah and University, Fez, Morocco 3Unity of GC/MS and GC, City of Innovation, Sidi Mohamed Ben Abdellah University, Fez, Morocco ARTICLE INFO ABSTRACT Article history: Essential oils are secondary metabolites of plants which have shown very important biological Received 16 October 2020 activities. The chemical composition, antioxidant and antimicrobial activity of essential oils Revised 15 November 2020 isolated from Haplophyllum tuberculatum (Forsskal) A. Juss are the subject of this study. The Accepted 20 December 2020 essential oils was extracted by hydrodistillation, GC/MS and GC-FID were used to determine Published online 02 January 2021 the chemical composition of the essential oils studied. Two tests were used to assess antioxidant power DPPH (2,2-diphenyl-1-picrilhidrazil) test and TAC (Total antioxidant capacity) assays. Agar diffusion and microdilution were used to examine the antimicrobial activity of these oils. The GC/MS and GC-FID analysis reveal 50 components. The major component was Farnesyl bromide (37.02%) followed by santolina triene (10.26%), terpinene-4-ol (6.13%) and limonene (2.26%). The scavenging capacity of DPPH by essential oils (IC = 3.2395 ± 0.009 mg/mL) was Copyright: © 2020 Agour et al. This is an open- 50 access article distributed under the terms of the lower than that of ascorbic acid (IC50= 2.8195 ± 0.008 μg/mL), but the measurement of its TAC revealed the presence of a significant amount of antioxidants in the oils studied (617.95 ± 0.589 Creative Commons Attribution License, which + permits unrestricted use, distribution, and mg EAA/g extract). S. aureus (bacteria G ) appears the most sensitive (24 mm). The values of reproduction in any medium, provided the original the MIC are proportional to the values of the diameters of the inhibition zone, the oil was very author and source are credited. active on S. aureus (MIC = 5.91μg/mL). E. coli and P. aeruginosa were the least sensitive (MIC = 94.6 μg/mL). The results of this study showed that this oil can be an alternative source of antimicrobials to combat strains with a problem of resistance to conventional antibiotics. Keywords: H. tuberculatum, Essential oils, GC/MS, Antioxidant, Antimicrobial activity. Introduction stings, throat inflammation, cough, such as dewormer, tonsillitis, otitis and loss of appetite. In Sudan the H. tuberculatum is used as an Medicinal plants have been used as a source of remedies antispasmodic, to treat allergic rhinitis and gynaecological disorders, since ancient times and the ancient Egyptians were familiar with many and on the cardiovascular System.10 Essential oils were used in ancient medicinal herbs and were aware of their usefulness in the treatment of 1 Rome, Greece and Egypt and throughout the Middle and Far East as various diseases. Interest in the use of herbal drugs has increased in 11 2 perfumes, food flavors, deodorants and pharmaceuticals . The both the traditional and modern medical systems. In addition, several essential oils which were utilized centuries ago in cosmetics usually clinical and preclinical studies reveal the biological effects of a wide 3 show interesting biological features. Essential oils are volatile range of plant-derived compounds. Haplophyllum tuberculatum aromatic concentrated hydrophobic oily liquids which are obtained (Forsskal) A. Juss is a species in the rutaceae family. This species is from various plant parts such as flowers, buds, seeds, leaves, twigs, one of 2100 species grouped into 154 genera composed of trees, bark, woods, fruits and roots.12 The rutaceae family contains many shrubs and aromatic herbs mainly tropical and subtropical 5 .This 6 aromatic and medicinal plants. Multiples studies have been reported family is a potential source of many medicinal substances. H. on the essential oils of H. Tuberculatum species belonging to different tuberculatum grows on sandy hills and rocky slopes of arid regions, it 13,14,15 7 regions in the world. Actually, however, essential oils and their extends from Morocco to the west of Pakistan, it has shown great components are gaining increasing interest because of their relatively variability in its traditional uses depending on geographic and 8 9 safe status, their wide acceptance by consumers, and their exploitation ecological location. According to Hadjadj et al., H. tuberculatum for potential multi-purpose functional use.16,17 According to Liaqat et was traditionally used as an antiseptic, calming, for infertility, al.,18 essential oils were used as antibacterial, antioxidant, antiviral, treatment of diabetes, against fever, bloating, diarrhea, flu, scorpion antifungal, anticancer, anti-inflammatory, antihistamine and anti- diabetic and in food preservation. Therefore, the determination of the *Corresponding author. E mail: [email protected] chemical composition of the volatile oil of H. tuberculatum harvest in Tel: +212 777801988 south-eastern Morocco, the evaluation of its antioxidant and antimicrobial power are the subject of this study. Citation: Agour A, Mssillou I, Saghrouchni H, Bari A, Lyoussi B and Derwich E. Chemical Composition, Antioxidant Potential and Antimicrobial Properties of the Essential Oils of Haplophyllum tuberculatum (Forsskal) A. Juss from Morocco. Trop J Nat Prod Res. Materials and Methods 2020; 4(12):1108-1115. doi.org/10.26538/tjnpr/v4i12.13 Plant material Official Journal of Natural Product Research Group, Faculty of Pharmacy, The aerial parts of H. tuberculatum were collected during the second University of Benin, Benin City, Nigeria. week of February 2019 near the town of Akka in the province of Tata (south-eastern Morocco). The plant has been identified by Pr. Amina 1108 © 2020 the authors. This work is licensed under the Creative Commons Attribution 4.0 International License Trop J Nat Prod Res, December 2020; 4(12):1108-1115 ISSN 2616-0684 (Print) ISSN 2616-0692 (Electronic) BARI botanist at the Sidi Mohamed Ben Abdellah University, Fez, Shimadzu 160-UV Spectrophotometer. Ascorbic acid was used as a Morocco. A voucher (RH001190122) sample was deposited at the positive control. The percentage of inhibition of free radicals DPPH herbarium of the Laboratory of Natural Substances, Pharmacology, was calculated by the following equation: Environment, Modeling, Health & Quality of Life, Faculty of Sciences Fez. PI (%) = (A0 - A/ A0) *100 Extraction of essential oils Where; PI: Percentage of inhibition. In order to extract the essential oils, the adopted method is A0: Absorbance of the control (containing all reagents except the test hydrodistillation using the Clevenger device.19 After their compound). fragmentation, 200 g of aerial parts pre-dried in the open air were A: Absorbance of the test compound. inserted into a 2 liter flask, 750 mL of water were added to it and the The IC50 is calculated graphically by linear regression of the graph. mixture was allowed to boil for 3 h. The essential oils obtained were stored at a temperature of -4°C. The yield (in percentage) was Determination of total antioxidant activity calculated on the basis of the dry weight of the vegetable materials. 200 µl of the essential oils (diluted 10 times in methanol) was combined with 2 mL of the reagent solution (0.6 M sulfuric acid, 28 Identification of essential oils components mM sodium phosphate and 4 mM ammonium molybdate), and the The analysis of essential oils of H. tuberculatum leaves was performed medium was incubated at 95 °C for 90 min. The optical density was by gas chromatography with flame-ionization detection (GC/FID) and measured at 695 nm in a Shimadzu 160-UV Spectrophotometer with a gas chromatography coupled with mass spectrometry (GC/MS), blank containing 200 μL of methanol in place of the extract. Ascorbic equipped with TRIPLUS AS S/N 20063460 in the light of the acid was used as standard, and the total antioxidant capacity was following experimental protocol, at the City of Innovation in Sidi expressed in milligrams of ascorbic acid per gram of dry extract (Mg Mohammed Ben Abdellah University, Fez. Morocco. EAA/ g of extract).22 Gas chromatography analysis (GC/FID) Antimicrobial activity evaluation The GC (Trace GC ULTRA, S/N 20062969, Thermo Fischer, France) Growing medium was equipped with flame ionization detector (GC-FID). The column For the disc diffusion method, the Muller Hinton Agar MHA medium used was, Varian capillary column (CP-Sil 5CB, 50m length, 0.32 mm was used for bacteria, and for yeast the Yeast Pepton Glucose YBG of diameter and Film thickness 1.25 µm). The column temperature medium was used. For the microdilution technique, the broths used was programmed from 40°C to 280°C for 5°C/min. The temperature are: YPG Broth for yeasts and Muller Hinton MHB Broth for bacteria. of
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