- Research Article
- Open Access
Chemical composition, antioxidant and antimicrobial potential of essential oils from different parts of Daphne mucronata Royle
© The Author(s) 2018
- Received: 9 July 2018
- Accepted: 21 November 2018
- Published: 17 December 2018
- D. mucronata
- Essential oil
Medicinal plants are well-known since beginning of human civilization for welfare of mankind and they dwell an imperative place in the socio-cultural as well as in the health-system of indigenous communities of Pakistan. Plant’s essential oils are worthwhile natural-products that are employed as raw materials in various fields, such as cosmetics, fragrances, phyto-therapy, nutrition and spices. Daphne mucronata Royle belongs to the family Thymelaeaceae. Common names of this plant include Kutilal, Nirko, Laighonai (laighuanay), Kheweshk. Leaves of this plant are poisonous and applied as insect repulsive abscesses for sore and glue is used for muscular and nerve troubles . Plant poultice is applied for rheumatism and sweeping . The plant has attractive flowers and can be used as decorative plant . The roots and shoots of D. mucronata Royle are considered as anthelmintic and employed in treatment of gonorrhea . Fruits are multipurpose so they are used for eating purposes and for treating eye problems, to cure skin, considered as remedy for face freckles, for killing lices, ticks and are also involved in coloring leather [4, 5]. Wood is used as firewood and used in preparation of gun powder charcoal . The bark is used in turmoil of bone for washing hairs and in folk medicines. Previous study revealed the presence of several phytochemicals, in this specie . To date, there are no previous reports related to Phyto-chemical composition as well as biological potential of plant Daphne mucronata Royle essential oils. As part of our efforts [8–12] this study is, therefore, reporting for the first time the aerial parts (stem and leaves) essential oil composition, and there biological potential.
Percentage yield and chemical composition of essential oils
GC/MS analysis of D. mucronata essential oils
7-Hexadecenoic acid, methyl ester, (Z)-
(Z,Z)-9,12-octadecadienoic acid methyl ester
11,14,17-Eicosatrienoic acid, methyl ester
Octadecanoic acid, methyl ester
Eicosanoic acid, methyl ester
1,2- diisooctyl benzenedicarboxylic acid ester
Behenic acid, methyl ester
Tetracosanoic acid, methy ester
Hexacosanoic acid, methyl ester
Antioxidant and antimicrobial potential of essential oils
% Yield and antioxidant analysis of D. mucronata Royle essential oils
Samples, standard compound
% Yield g/100 g
% Inhibition of peroxidation in linoleic acid
DPPH radical scavenging IC50 (µg/mL)
Leaves essential oil
37.57 ± 0.89
85.15 ± 0.31
Stem essential oil
64.16 ± 0.93
45.46 ± 0.04
89.1 ± 0.78
9.01 ± 0.10
Antimicrobial activity of D. mucronata Royle essential oils
Leaves essential oil
Stem essential oil
Zone of inhibition (mm)
Zone of inhibition (mm)
Zone of inhibition (mm)
19.0 ± 0.60
0.86 ± 0.001
20.7 ± 0.55
0.48 ± 0.001
21.7 ± 0.49
0.97 ± 0.0003
18.9 ± 0.20
2.44 ± 0.002
22.2 ± 0.01
0.98 ± 0.005
23.8 ± 0.67
0.25 ± 0.0001
8.88 ± 0.01
11.2 ± 0.40
25.26 ± 0.3
0.46 ± 0.0002
22.9 ± 0.43
0.39 ± 0.0007
30.0 ± 0.32
0.25 ± 0.0001
23.4 ± 0.50
0.33 ± 0.0003
The entire plant “D. mucronata Royle” was attained from Quetta, Pakistan. The plant was identified by Prof. Dr. Rasool Bakhsh Tareen, Botany Department, University of Balochistan, Quetta, Pakistan, where we deposited sample-specimen (Voucher # DM-RBT-09).
Essential oil extraction
For the essential oils extraction, 50 g of each part (stem and leaves) of powdered plant materials dried under the shady place, were hydro distillated by employing a Clevenger-type device for 5 h. Sodium sulphate (Na2SO4) was used for drying the extracted essential oils, then after filtration oils were stored in a vial at 4 °C till start of further analysis.
TR(x) = represents retention time of compounds to be identified
TR(n) = retention time of carbon (Cn)
TR(n+i) = retention times of carbon (Cn+i)
DPPH radical scavenging assay
Percentage-inhibition of linoleic peroxidation
Analysis of reducing power
At different concentrations (2.5–10 mg), the plant oils were mingled with 1% potassium ferricyanide (5 mL) and 5 mL of sodium phosphate buffer (0.2 M, pH 6.6) solution. For 20 min at 50 °C, the reaction mixture was heated and after that, 10% of trichloroacetic acid (5 mL) was mixed with heated reaction mixture. Then the resultant solution was subjected for centrifugation for 10 min at 5 °C at the rate of 980 rpm. At that time, the 5 mL of upper layer of reaction mixture was dissolved in 5 mL of distilled H2O. As a final point, 1 mL of 0.1% freshly prepared FeCl3 solution was added in it. At 700 nm absorbance was noted and result were obtained in triplicates .
Four different bacteriological strains (Bacillus cereus ATCC 14579, Escherichia coli ATCC 25922, Staphylococcus epidermidis ATCC 12229 and Nitrospira moscoviensis locally isolated) and three different fungal strains (Aspergillus niger ATCC 10595, Candida albicans ATCC 10231, Aspergillus flavus ATCC 32612) were used to check the antimicrobial effects of essential oils. For this study, pure microbial organisms were provided by Department of Veterinary Microbiology (DVM) (University of Agriculture Faisalabad (UAF), Pakistan). The nutrient agar was employed to culture bacteriological strains overnight at 37 °C while potato dextrose agar (PDA) was cast off for the development and culturing of fungal strains at 28 °C.
Disc diffusion method
The antimicrobial potential of plant essential oils was determined by Disc Diffusion method . For this, the 6 mm diameter discs were employed whose soaking was performed with 20 mg/mL essential oil (100 μL/disc). Moreover, soaked disk were placed on the inoculated agar. Discs without samples were used as negative control. The fungone (100 μL/disc) and Rifmapicin (100 μL/disc) were served as a positive control for fungal and bacteriological strains respectively. The incubation of petri-dishes for bacteria were performed at 37 ± 0.1 °C for 24 h while for fungi at 28 ± 0.3 °C for 48 h. For the results, zones of inhibition (ZOIs) formation were measured on the agar media.
Minimum inhibitory concentration (MIC)
The resazurin microtitre-plate assay was employed to determine the minimum inhibitory concentration (MICs) of the D. mucronata essential oils .
All samples were analyzed in triplicate. Data were analyzed by analysis of variance (ANOVA) using Costat (Version 3.8) statistical software.
We have investigated essential oils from aerial parts of Daphne mucronata obtained by hydro-distillation process. Fifty-one different compounds were found in stem and leaves essential oils by GC–MS analysis. These compounds made the essential oils very effective in antimicrobial and antioxidant potential. Our study revealed that oils obtained from D. mucronata could be a promising source of effective antioxidant and antimicrobial compounds and may play vital role for discovery of new drugs against pathogenic diseases. Both of these essential oils may play an important role in flavoring and cosmetic industry.
IA, MZ, KR, NR and MJ made a significant contribution to Conceptualization, data curation and experimental work. SAK, RBT contributed towards formal analysis. VUA, AM, MR, MZUH and HZEJ contributed to interpretation of data and helped in drafting of manuscript. All authors read and approved the final manuscript.
The authors are thankful to Higher Education Commission Pakistan (HEC) for funding through the Research Project No 20-1563/R&D/09/1582.
The authors declare that they have no competing interests.
Availability of data and materials
All the main experimental and characterization data have been presented in the form of tables and figures.
Consent for publication
We the all authors consent to publication.
Ethics approval and consent to participate
The research was funded by Higher Education Commission (HEC), Pakistan.
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