Study of Chemical and Biological Aspects of Valeriana Wallichii DC
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Asian Journal of Chemistry; Vol. 24, No. 7 (2012), 3243-3246 Study of Chemical and Biological Aspects of Valeriana wallichii DC. Root Essential Oil * ZAHIDA PARVEEN , SAIMA SIDDIQUE, MUAFIA SHAFIQUE, SHAISTA JABEEN KHAN and RAZIA KHANUM PCSIR Laboratories Complex, Ferozpur Road, Lahore-54600, Pakistan *Corresponding author: Fax: +92 42 99230705; Tel: +92 42 99230688-95 Ext: 318; E-mail: [email protected] (Received: 15 September 2011; Accepted: 24 February 2012) AJC-11113 The essential oil of Valeriana wallichii DC. root, which is an indigenous plant was extracted by hydro distillation. Chemical composition of the oil by GC-MS analysis revealed the presence of seven new components i.e. isoaromadendrene epoxide (3.435 %), isocaryophyllene (1.919 %), epiglobulol (1.692 %), longifolene-[V4] (1.618 %), panasinene (1.452 %), lognifolenaldehyde (0.875 %) and butanoic acid, 3-methyl-, 3-methylebutylster (0.840 %). Patchouli alcohol (16.425 %) was found to be the major component among ten identified components of the essential oil. Antibacterial activity and antioxidant activity of the said essential oil was investigated for the first time. The antibacterial potential of the essential oil of V. wallichii was determined by paper disc diffusion method, against Gram positive and Gram negative bacteria. Essential oil without any dilution was found to be more effective against all test microbes as compared to the standard antibiotics i.e. streptomycin and penicillin G. used as positive controls. DPPH scavenging effect was 96.47 % for essential oil of V. wallichii. Antioxidant activity of the oil was 11.27 % higher than BHT used as control. Key Words: Valeriana wallichii, Valerianaceae, Essential oil, Gas chromatography, Antibacterial activity, Antioxidant activity, Patchouli alcohol. INTRODUCTION reported to be between 0.2 and 0.7 %1. They are used in perfumes, cosmetics, soap and other products, for flavouring Valeriana wallichii DC (Asian specie) belongs to family food and drink and for scenting incense and household Valerianaceae, has about 200 perennial herb and small shrub cleaning products5,7. The antimicrobial and antioxidant activity species throughout the world, is chiefly found in the temperate of essential oils have great medicinal as well as food value. Himalaya and cold regions of the northern hemisphere. Other Increase in the emergence of new bacterial strains that commonly used specie of genus valeriana include V. officinalis are multi-resistant, coupled with the non-availability and high 1 from Eastern Europe and V. edulis from Americas . V. wallichii cost of new generation antibiotics have resulted in increase is an indigenous plant from Indian subcontinent used since long morbidity and mortality8. Hence, there is need to look for potent 2 in Ayurvedic and Unani systems of medicine . Valeriana antimicrobial and antioxidant agents from other sources. So, wallichii DC is a small perennial herb of 14-45 cm height, with investigations of the antimicrobial activities, mode of action root stock, thick branching stem, sharply pointed leaves, white and potential uses of plant essential oils have regained or pink flowers in clusters and hairy fruit. It occurs in Kashmir, momentum. Studies showed that most of the essential oils 3,4 Muree hills, Punjab and Northern areas of Pakistan . Valerian showed significant antimicrobial activity7. Literature indicates is widely used as a mild sedative and sleep aid for insomnia, that studies on V. wallichii are scarcely found. Recently, excitability and exhaustion. Other medicinal uses include essential oil of V. wallichii is reported to show the antifungal9 diuretic, carminative antispasmodic, anticonstipation, anti- and nematocidal activity10. However, its antibacterial and 5 scorpion poison and to improve blood circulation . antioxidant activities are not yet reported. Essential oils and extracts obtained from plants have recently The aim of present study is to determine the chemical gained a great popularity and scientific interest because of their composition, antimicrobial and antioxidant properties of relatively safe status, wide acceptance by consumers and exploi- essential oils from V. wallichii an indigenous Indian subconti- 6 tation for potential multi-purpose functional use . Essential oils nent plant specie. are concentrated, hydrophobic liquids containing volatile aroma compounds from plants. Various essential oils have been used EXPERIMENTAL medicinally at different periods in history. Valerian oil is obtained Extraction of essential oil: The V. wallichii roots were by steam distillation of the dried, ground roots and yield is collected from the local market. They were cleaned from 3244 Parveen et al. Asian J. Chem. extraneous matter. The essential oil was extracted through to examine the antioxidant activity of V. wallichii oil. The hydro-distillation by reverse Dean Stark assembly11. The steam scavenging effect on DPPH radical was determined by distillate was removed, dried over anhydrous sodium sulphate modifying the previous methods15. Briefly, 0.004 % DPPH and stored at 4 ºC for further studies. solution was prepared in methanol. The essential oil was mixed GC-MS analysis: The analysis of the essential oil was with methanol in appropriate amounts to prepare 20, 40, 60, carried out on GC-MS of Agilent Technologies, Model 6890N, 80 and 100 % ( v/v) methanolic test solutions. Butylated operating in EI mode at 70 ev equipped with a split-splitless hydroxytoluene (BHT) was used as a positive control. A injector. Helium used as a carrier gas at the flow rate of 1 mL/ butylated hydroxytoluene stock solution (100 µg/mL) was used min, while HP-5MS (30 m × 0.25 mm id, 0.25 µ film thickness) to prepare 20, 40, 60 and 80 %(v/v) methanolic solution, capillary column was used. The initial temperature was whereas butylated hydroxytoluene stock solution (100 µg/mL) programmed at 50-140 ºC at the rate of 5 ºC/min and then was considered as 100 % working solution. Each methanolic 100-250 ºC at the rate of 3 ºC/min followed by a constant dilution of essential oil as well as butylated hydroxytoluene temperature at 260 ºC for period of 20 min. Sample (2 µL) (100 µL) was mixed with 3 mL of DPPH solution separately. was injected to column programmed at 200 ºC and resolutions The mixtures were shaken vigorously and left to stand for 0.5 of components were attained. The components were identified h in dark at room temperature. The absorbance of the resulting by their retention time and peak enhancement with standard solutions was measured at 517 nm using a UV-VIS spectro- samples in gas chromatographic mode and NIST library search photometer (Nicolet, Evlution-300, Germany). Decreasing from the derived fragmentation pattern of the various compo- absorbance of DPPH solution indicates an increase in DPPH nents of the oil. radical scavenging activity corresponding to the antioxidant Antibacterial assay: In vitro antimicrobial studies were activity of the experimental V. wallichii essential oil. This carried out on eight bacterial strains including Bacillus subtilis activity is given as % inhibition, which is calculated with the ATCC6333, Klebsiella pneumoniae, Salmonella typhimurium, following equation: Pseudomonas aeruginosa, Pseudomonas fluorescens, Staphy- DPPH scavenging effect (% inhibition) = {(Ablank - Asample)/ lococcus aureus, Escherichia coli and Enterobacter aerogenes. Ablank} × 100 Among the tested microorganisms Bacillus subtilis ATCC6333 where, A stands for absorbance. Each experiment was perfor- was obtained from microbiology laboratory of PCSIR labs med in triplicate. complex Lahore and other were collected from pathological laboratory of a local hospital. All clinical isolates were charac- RESULTS AND DISCUSSION terized to specie level according to standard microbiological The essential oil was extracted by hydrodistillation from 12 techniques described by Monica . The cultures of bacteria the V. wallichii roots, having 0.245 % (v/w) yield on dry weight were maintained in the laboratory on nutrient agar slants at basis. Present results indicate that the essential oil yield of V. 4 ºC throughout the study. wallichii roots (0.245 %) is higher than the claim made by 13 The paper disc diffusion method was applied with slight Bos et al.16 (0.09 %) on dry weight basis. The gas chromato- modification to test the antimicrobial activity of V. wallichii graphy coupled with mass spectrometric analysis revealed the essential oil. Normal strength nutrient agar medium (OXOID, presence of 26 compounds, out of which 10 components were England) was prepared and autoclaved at 121 ± 1 ºC for 15 identified (Table-1). These components were further classified min. For antibacterial assay 24 h old bacterial cultures grown in two fractions i.e. hydrocarbon fraction and oxygenated -1 at 37 ºC were used. Cultures were diluted 10 in sterile ringer fraction. Hydrocarbon fraction constituted camphene, 14 6 solution to set an inoculums density of approximately 10 isocaryophillene, isocaryophillene, panasinene while butanoic CFU/mL, which was used for the test. 30 µL of these suspen- acid, 3-methyl-, 3-methylbutylester, bornyl acetate, sions were inoculated to plates containing sterile nutrient agar epiglobulal, patchouli alcohol, isoarmadendrene epoxide, medium using a sterile cotton swab. lognifolenaldehyde comprized the oxygenated fraction of the Each filter paper discs (6 mm in diameter) impregnated oil. Presence of patchouli alcohol (16.425 %) as major compo- with 25 µL of different concentrations