- Research article
- Open Access
Compositional studies and Biological activities of Perovskia abrotanoides Kar. oils
https://doi.org/10.1186/0717-6287-47-12
© Ashraf et al.; licensee BioMed Central Ltd. 2014
- Received: 24 October 2013
- Accepted: 31 December 2013
- Published: 1 April 2014
Abstract
Background
Current study has been designed to evaluate the chemical composition of essential and fixed oils from stem and leaves of Perovskia abrotanoides and antioxidant and antimicrobial activities of these oils.
Results
GC-MS analysis of essential oil identified 19 compounds with (E)-9-dodecenal being the major component in stem and hexadecanoic acid in leaves. In contrast, GC-MS analysis of fixed oil showed 40 constituents with α-amyrin the major component in stem and α-copaene in leaves. The antioxidant activity showed the highest value of 76.7% in essential oil from leaves in comparison with fixed oil from stem (45.9%) through inhibition of peroxidation in linoleic acid system. The antimicrobial assay tested on different microorganisms (e.g. E. coli, S. aureus, B. cereus, Nitrospira, S. epidermis, A. niger, A. flavus and C. albicans) showed the higher inhibition zone at essential oil from leaves (15.2 mm on B. cereus) as compared to fixed oil from stem (8.34 mm on S. aureus) and leaves (11.2 mm on S. aureus).
Conclusions
The present study revealed the fact that essential oil analyzed from Perovskia abrotanoides stem and leaves could be a promising source of natural products with potential antioxidant and antimicrobial activities, as compared to fixed oil.
Keywords
- Perovskia abrotanoides
- Essential oil
- Fixed oil
- Antioxidant capacity
- Antimicrobial activity
Introduction
Perovskia abrotanoides (Lamiaceae) locally known as hoosh, visk, brazambal, domou, and gevereh[1–4] is a medicinally important plant found in Baluchistan province and northern areas of Pakistan. The plant is used by local communities for treatment of typhoid, headache, gonorrhea, vomiting, motion, toothache, atherosclerosis, cardiovascular diseases, liver fibrosis, and cough [5–7]. It has sedative, analgesic, antiseptic and cooling effect [5, 8, 9]. Herbal tea of this plant is used in curing infection problems and painful urination [10].
Some of the pharmacological effects of plant such as antiplasmodial, antiinflammatory and cytotoxic effects have also been reported [11–13]. Its antioxidant performance including heart enhancing and optimized performance as cell toxicity in pathogens, viruses and cancer cells were also reported [5, 14]. Essential and fixed oils of Perovskia abrotanoides plays an important role in protection of stored grains and showed to be effective in washing wounds, anti-ring worm, dermal parasites, anti-fungus and anti-hypoxia [14–17]. Despite its multipurpose usage, little data exists on chemical composition as well as biological activities of this plant. Therefore this study was designed to investigate composition of essential and fixed oil and antioxidant and antimicrobial activities of stem and leaves of Perovskia abrotanoides.
Results and discussion
The diversity of agrogeoclimatic conditions of Pakistan offers the broadest array of flora. This rich floral biodiversity of Pakistan constitutes an impressive pool for ‘natural food and healing’ from which the indigenous communities select ingredients for food and prepare herbal recipes for the treatment, management and control of their various ailments. It is believed that antimicrobials and antioxidants of plant origin are without side effects as compared to synthetic drugs and have an enormous therapeutic potential to heal many infections and diseases. Much of the potential of these botanicals is however still unearthed. In current research, a less explored medicinal plant found in Pakistan has been investigated for compositional studies and biological activities.
GC-MS analysis of essential and fixed oils
GC-MS product distribution of the P. abrotanoides essential oil from stem (top) compared with essential oil from leaves (bottom).
GC-MS product distribution and relative proportion (wt% of total compounds) of dry weight essential oil (stem and leaves) from Perovskia abrotanoides (in mg/g oil)
RI | Chemical constituents | % Composition | |||
|---|---|---|---|---|---|
Essential oil | Fixed oil | ||||
Stem | Leaves | Stem | Leaves | ||
820 | 2,2,3,4-tetramethylpentane | 0.46 | -- | -- | -- |
850 | t* | -- | 0.77 | -- | -- |
922 | 2,4,6-trimethyloctane | 0.76 | -- | -- | -- |
973 | Nonane, 5-(2-methylpropyl) | -- | 1.07 | -- | -- |
1000 | t* | -- | -- | 0.33 | -- |
1033 | Eucalyptol | -- | -- | -- | 1.26 |
1052 | 2,2,5,5- tetramethylhexane | 3.96 | -- | -- | -- |
1125 | Ethylbenzene | -- | -- | -- | 3.08 |
1143 | Camphor | -- | -- | -- | 1.54 |
1165 | Borneol | -- | -- | -- | 2.12 |
1175 | S-propyl methanesulfonothioate | 3.0 | -- | ||
1189 | α terpineol | -- | -- | -- | 1.07 |
1200 | 1-bromo dodecane | -- | -- | 0.11 | -- |
1236 | 2-pentylfuran | -- | -- | 0.52 | 3.05 |
1291 | Bornyl acetate | -- | -- | -- | 0.7 |
1352 | Terpinyl acetate | -- | -- | -- | 1.26 |
1362 | Geranyl acetate | -- | -- | -- | 0.83 |
1399 | Tetradecane | 2.41 | 5.19 | -- | -- |
1403 | (E)-9-dodecenal | 66.5 | -- | -- | -- |
1404 | δ-caryophyllene | -- | -- | -- | 0.7 |
1416 | α-copaene | -- | -- | -- | 10.99 |
1424 | Caryophyllene | t* | -- | -- | 0.95 |
1454 | 1,4,7-cycloundecatriene,1,5,9,9-tetramethyl, Z,Z,Z- | -- | -- | -- | 0.82 |
1500 | Pentadecane | -- | 3.97 | t* | -- |
1532 | Epiglobulol | -- | -- | 0.65 | 4.95 |
1642 | Cubenol | -- | -- | -- | 1.64 |
1654 | α-cadinol | -- | -- | 0.48 | -- |
1659 | β-eudesmol | -- | -- | 0.42 | -- |
1719 | 8-hexylpentadecane | -- | -- | -- | 1.16 |
1720 | Tetradecanoic acid | -- | -- | 0.17 | -- |
1729 | 4-(E)-3-hydoxyprop-1-enyl)-2-methoxyphenol | -- | -- | 1.26 | 1.93 |
1750 | t* | -- | -- | 0.33 | -- |
1800 | t* | -- | -- | -- | 0.81 |
1800 | Cis-9-hexadecenal | -- | -- | 2.23 | -- |
1822 | 5,6-dimethylheptadecane | 2.94 | -- | -- | -- |
1834 | Unidentified | -- | -- | 2.61 | 2.06 |
1847 | 2-pentadecanone, 6,10,14-trimethyl | -- | -- | -- | 0.79 |
1854 | 5-methyloctadecane | -- | -- | 0.48 | 3.21 |
1914 | Anthranilic acid | -- | -- | -- | 1.04 |
1949 | Heptadec-1-ol | -- | -- | -- | 0.99 |
1962 | Hexadecanoic acid, methyl ester | -- | 27.79 | -- | -- |
2000 | Eicosane | -- | 6.22 | -- | -- |
2000 | Unidentified | -- | -- | -- | 5.63 |
2010 | Unidentified | -- | -- | -- | 1.28 |
2012 | Isopropyl hexadecanoate | -- | -- | 6.56 | 6.67 |
2080 | Octadecanoic acid, 17- methyl ester | -- | 2.19 | -- | -- |
2088 | Octadecanoic acid, methyl ester | 8.37 | 18.45 | -- | -- |
2100 | Unidentified | 1.93 | -- | -- | -- |
2110 | t* | -- | -- | -- | 0.69 |
2111 | Trans phytol | -- | -- | 2.03 | 7.33 |
2138 | 7-methoxy-8-(3-methyl-2-butenyl)-2H-chromen-2-one | -- | -- | 2.29 | -- |
2161 | Oleic acid | -- | -- | -- | 3.31 |
2200 | Stearic acid | -- | -- | 0.33 | -- |
2221 | Nonadecanoic acid | -- | -- | 0.6 | -- |
2240 | Androst-2-en-17-one | -- | -- | 2.18 | --- |
2330 | Trans-ferruginol | -- | -- | 0.48 | 3.06 |
2400 | Tetracosane | -- | -- | 0.17 | -- |
2430 | 3-hydroxyandrostan-17-one | -- | -- | -- | 2.06 |
2435 | t* | -- | -- | -- | 0.88 |
2526 | Docosanoic acid | -- | 2.47 | -- | -- |
2700 | Unidentified | -- | 1.1 | -- | -- |
2730 | Tetracosanoic acid | -- | 2.62 | -- | -- |
3011 | t* | 0.53 | -- | -- | |
3098 | 5.β-cholestan-3α-ol | -- | 1.46 | -- | -- |
3400 | Tetratriacontane | -- | -- | 0.14 | -- |
3408 | β sitosterol | -- | -- | 2.82 | -- |
3654 | 1-(+)-Ascorbic acid 2,6-dihexadecanoate | 3.0 | -- | -- | -- |
3654 | Unidentified | -- | -- | -- | 3.3 |
3600 | Hexatriacontane | -- | 0.69 | -- | -- |
4000 | Unidentified | 2.51 | 1.99 | -- | -- |
4050 | α-amyrenone | -- | -- | 11.8 | 5.19 |
4160 | α-amyrin | -- | -- | 47.01 | -- |
4175 | Lupeol | -- | 21.5 | -- | -- |
Antioxidant activity
% Yield and Antioxidant activity of Perovskia abrotanoides essential and fixed oils*
Plant parts and tested samples | % yield (g/100 g) | DPPH radical scavenging (IC50) (μg/mL) | Inhibition of peroxidataion in linoleic acid system (%) | |
|---|---|---|---|---|
Stem | Essential oil | 2.13 ± 0.06 | 17.9 ± 0.004 | 61.1 ± 0.87 |
Fixed oil | 2.96 ± 0.04 | 73.1 ± 0.007 | 45.9 ± 1.74 | |
Leaves | Essential oil | 2.86 ± 0.05 | 45.4 ± 0.01 | 76.7 ± 0.87 |
Fixed oil | 3.61 ± 0.10 | 62.5 ± 0.05 | 47.4 ± 0.87 | |
Standard | BHT | -- | 8.78 ± 0.10 | 90.4 ± 0.87 |
The percent inhibition of linoleic acid peroxidation was observed for plant essential and fixed oils whereas synthetic BHT provided inhibition at the level of 90.4%. Essential oil of leaves exhibited highest % inhibition of peroxidation (76.4%) followed by stem essential oil (61.1%), leaves fixed oil (47.4%) and stem fixed oil (45.9%). When the results of DPPH scavenging activity and the percent inhibition of linoleic acid oxidation were compared with standard BHT, all the oils showed significantly (p < 0.05) minor activity. Essential oils have the efficacy to reduce the peroxide formation during incubation in linoleic acid system as per our previous studies [25–27].
Reducing potential of essential and fixed oils of P. abrotanoides (stem and leaves).
Antimicrobial activity
Antimicrobial activity of Perovskia abrotanoides essential and fixed oils*
Diameter of Inhibition Zone (IZ, mm) | ||||||
|---|---|---|---|---|---|---|
Tested microrganisms | Stem | Leaves | Standard drugs | |||
Essential oil | Fixed oil | Essential oil | Fixed oil | Novidate | Fungone | |
E. coli | - | 12.9 ± 0.03 | - | - | 27.7 ± 0.02 | - |
S. aureus | - | 8.34 ± 0.03 | - | 11.2 ± 0.04 | 21.2 ± 0.03 | - |
B. cereus | - | - | 15.2 ± 0.05 | - | 24.5 ± 0.02 | - |
Nitrospira sp. | 9.76 ± 0.01 | - | - | - | 31.7 ± 0.04 | - |
S. epidermidis | - | - | - | 14.2 ± 0.07 | 23.4 ± 0.01 | - |
A. niger | - | - | 10.6 ± 0.005 | 8.46 ± 0.01 | - | 30.2 ± 0.06 |
A. flavus | 9.94 ± 0.03 | - | - | 9.64 ± 0.02 | - | 28.4 ± 0.01 |
C. albicans | - | 8.07 ± 0.02 | 9.78 ± 0.01 | 8.32 ± 0.03 | - | 30.2 ± 0.01 |
Minimum Inhibitory Concentration (MIC, mg/mL) | ||||||
E. coli | - | 7.95 ± 0.02 | - | - | 0.77 ± 0.01 | - |
S. aureus | - | 14.2 ± 0.03 | - | 9.40 ± 0.05 | 1.06 ± 0.02 | - |
B. cereus | - | - | 5.75 ± 0.04 | - | 0.95 ± 0.03 | - |
Nitrospira sp. | 19.6 ± 0.05 | - | - | - | 0.28 ± 0.01 | - |
S. epidermidis | - | - | - | 5.25 ± 0.02 | 0.88 ± 0.03 | - |
A. niger | - | - | 11.4 ± 0.02 | 18.4 ± 0.01 | - | 0.47 ± 0.02 |
A. flavus | 14.5 ± 0.01 | - | - | 17.1 ± 0.03 | - | 0.24 ± 0.01 |
C. albicans | - | 1.26 ± 0.01 | 11.7 ± 0.08 | 1.93 ± 0.02 | - | 0.17 ± 0.02 |
Conclusions
We have investigated two different types of Perovskia abrotanoides oils. The presence of different compounds founded by GC-MS analysis, rendered the essential oil very efficient in antioxidant and antimicrobial capacity in respect with fixed oil which could be due to the presence of unsaturated fatty acids and anthocyanins compounds. We believe that the compounds, in particular (E)-9-dodecenal and hexadecanoic acid from essential oil are directly involved in antioxidant and antimicrobial processes. Finally, our study revealed the fact that essential oil analysed from Perovskia abrotanoides stem and leaves could be a promising source of natural products with potential antioxidant and antimicrobial activities in respect with fixed oil.
Methods
Collection of plant material
The Perovskia abrotanoides whole plant was collected and deposited from Quetta and Ziarat Valley and further identified by Dr. Rasool Bukhsh Tareen, Department of Botany, University of Baluchistan, Quetta, Pakistan.
Essential oil extraction
Dried powdered of plant material (100 g) was subjected to hydro-distillation for 5 h using a Clevenger type apparatus for extraction of stem and leaves from essential oils. The extracted essential oil was dried over anhydrous Na2SO4, filtered and stored in a vial at 4°C until further analysis.
Fixed Oil extraction
The fixed oil was extracted following the method of Ajayi and contributors [32]. Briefly, 100 g of the shade dried powdered stem and leaves of plant were separately extracted in 250 mL n- hexane (99.9% purity) solvent by using a Soxhlet extractor for 6 h and then the extra-solvent was removed by distillation under reduced pressure in a rotary evaporator at 35°C and the pure oil was kept at 4°C in the dark.
GC-MS analysis of essential and fixed oils
Essential and fixed oils were analysed by GC-MS (QP2010, SHIMADZU, Japan) using an Agilent GC 6890 N model (1 μL sample injected, split 1:50 column flow 1.0 mL/min., program temp. 200°C, rate 10°C/min) coupled with a quadrupolar MS 5973. GC was equipped with capillary column (30 m × 0.25 mm; film thickness 0.25 μm). Oven temperature was kept at 45°C first for 5 min, and then raised at 325°C at a 15°C/min for another 5 min. Helium gas was then employed at a flow rate of 1.1 mL/min (60 KPa pressure; 38.2 cm/sec linear velocity). The identification of components was based on comparison of their mass spectra with those of NIST mass spectral library [33].
Antioxidant activity
The 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay
Where:
Ablank = absorbance of the control
Asample = absorbance of the test samples
Inhibition of linoleic acid peroxidation
Analysis of reducing power
The reducing power of the oils was determined according to the procedure described by Yen et al. [36] with little modification. The plant oils at various concentrations (2.5-10 mg) were mixed with sodium phosphate buffer (5 mL; 0.2 M) of pH 6.6 and potassium ferricyanide (5 mL; 1.0%). The mixture was heated for 20 min at 50°C. Then trichloroacetic acid (5 mL; 10%) was added and centrifuged at 980 rpm at 5°C for 10 min. Further, its upper layer (5 mL) was dissolved in 5 mL distilled water and finally freshly prepared ferric chloride (1 mL; 0.1%) was added. The absorbance was calculated at 700 nm and a result for each sample was recorded in triplicate.
Antimicrobial assay
Microbial strains
Bacillus cereus ATCC 14579, Escherichia coli ATCC 25922, Nitrospira sp. locally isolated, Staphylococcus epidermidis ATCC 12229, Staphylococcus aureus API Staph tac 6736153 were used as bacterial strains and Aspergillus niger ATCC 10595, Aspergillus flavus ATCC 32612, Candida albicans ATCC 10231 as fungal strains. The pure bacterial and fungal strains were obtained from the Department of Veterinary Microbiology, University of Agriculture, Faisalabad, Pakistan. The bacterial strains were cultured overnight at 37°C in nutrient agar (Oxoid, UK) while fungal strains were cultured overnight at 28°C using potato dextrose agar (Oxoid, UK).
Antimicrobial disc susceptibility test
Antimicrobial activity of the plant oils was determined by using the disc susceptibility test [37]. The discs (6 mm diameter) were impregnated plant oils (100 μL/disc) placed aseptically on the inoculated agar. Discs without injected samples served as a negative control, and Novidate (100 μL/disc) and Fungone (100 μL/disc) (both from Oxoid, UK) as positive control. The Petri dishes were incubated at 37 ± 0.1°C for 20–24 h and 28 ± 0.3 for 40–48 h for both bacteria and fungi. At the end of the period, the inhibition zones were measured. The positive antimicrobial activity was read based on growth inhibition zone.
Minimum inhibitory concentration (MIC)
The MIC of the plant oils was estimated following resazurin microtitre-plate assay reported by Sarker and contributors [38].
Statistical analysis
All the aforementioned experiments were conducted in triplicate. Statistical comparisons were performed by one-way analysis of variance (ANOVA) followed by Dunnett’s t-test using SPSS version 12.0 (SPSSS Inc., Chicago, IL, USA). Probability values < 0.05 were considered to indicate significant difference.
Declarations
Authors’ Affiliations
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