RHIZOME ESSENTIAL OIL COMPOSITION OF ZINGIBER CERNUUM AND ITS ANTI MICROBIAL ACTIVITY

RHIZOME ESSENTIAL OIL COMPOSITION OF ZINGIBER CERNUUM AND ITS ANTI MICROBIAL ACTIVITY          

Mity Thambi ^*1 and Mohamed P Shafi ²

Department of Chemistry ¹, Catholicate College, Pathanamthitta, Kerala, India.

Department of Chemistry ², University of Calicut, Kerala, India.

ABSTRACT: Zingiber cernuum belongs to family Zingiberaceae is a perennial herb, also called as Curved stem ginger. The rhizomes of Zingiber cernuum is steam distilled and the composition of essential oil was studied. The essential oil obtained was yellow in colour and the yield was about 0.007%. The chemical profile of the oil was obtained from their GC-MS analysis. The major components present in the oil are trans-Caryophyllene, α-Humulene and δ-3-Carene. The antimicrobial activity of the essential oil was studied using disc diffusion method. The essential oil was tested against four pathogenic stains of gram positive and gram negative bacteria. The essential oil of Zingiber cernuum show considerable antibacterial activity. The oil was found to be active against both gram positive and gram negative bacteria. The anti bacterial activity is mainly due to the major components present in the oil or due to the synergistic effect of the major and minor components.

Zingiber cernuum,

 GC-MS, Disc-diffusion method

INTRODUCTION: Zingiber cernuum Dalzell Common name: Curved-Stem Ginger, Nodding-stem ginger. Family: Zingiberaceae (Ginger family) Curved-Stem Ginger is a large perennial herb, 1-2 m tall, with curved stem. Leaves are 15-30 cm, narrow-elliptic, long-pointed. Flowers are borne in spikes 5-10 cm long, directly from the rootstock, rising just above the ground. Bracts are 2-3 cm long, greenish-yellow. Sepal cup is shortly 3-lobed. Stamen is one with short filament. Style is threadlike. Capsules are 1 cm long, smooth with red, channeled seeds. Flowers are creamish, variegated with red, with the lib broad and 3-lobed.

Flowers open at night. Curved-Stem Ginger is found in the evergreen forests of Western Ghats, India. Kulkarni and coworkers identified flavanoid and tannins from the phytochemical investigation of Z. cernuum ¹. This species shows an appropriate amount of iron, manganese and low amount of molybdenum, sulphur, nitrate in rhizome and leaf ². Phytochemical screening of Z. cernuum shows flavnoids which shows antioxidant activity ³.

In the present investigation the composition of essential oil from the rhizome of Z.cernuum was studied and the chemical profiles of the oil were obtained from their GC-MS analysis.

MATERIALS AND METHODS:

Extraction: The plant material was collected from Malappuram district, Kerala, India. The rhizome (Z.cernuum-1.1kg) was ground into a paste and subjected to steam distillation separately for 5 hours. The oils were extracted with diethyl ether. The ether extract was dried using anhydrous sodium sulphate and ether evaporated. The pure oil weighed 80 mg.

Analysis of the oil:

GC-FID analysis was carried out using a Perkin Elmer Clarus 500 GC equipped with a 30 m × 0.32 mm Elite-5MS capillary column (0.32 μm film thickness). 1 μl of each sample was diluted with 300 μl of Et₂O and injected (0.5 μl) in the “split” mode (1:30) with a column temperature programme of 40°C for 5 min, then increased to 280°C at 4°C/min and finally held at this last temperature for 10 min. Injector and detector were set at 250 and 300°C, respectively, and the carrier gas was He with a head pressure of 12.0 psi.

GC/MS analyses were carried out using a Perkin Elmer Clarus 500 GC equipped with a Clarus 500 mass spectrometer using the same capillary column and chromatographic conditions as for the GC/FID analysis. Mass spectra were acquired over 40-500 amu range at 1 scan/sec. The identification of essential oil components was performed by means of their retention indices (RI), by a peak matching library search ⁴ and by comparison with authentic reference compounds as well as with published mass spectra ^{5, 6}. Retention indices (RI) were calculated using a n-alkane series (C₆-C₃₅) under the same GC conditions as for the samples. The relative amount (%) of individual components of the oil is expressed as percent peak area relative to total peak area from the GC/FID analyses of the whole components.

Anti microbial activity:

The antibacterial screening of the extract was carried out by determining the zone of inhibition using standard method ⁷. The essential oil was tested against four pathogenic bacterial strains of gram positive and gram negative organism by disc diffusion method ⁸. The test microorganisms of gram positive bacteria:   Staphylococcus aureus, Bacillus subtilis, Streptococcus faecalis, Staphylococcus albus. gram negative  bacteria: Escherichia  coli, Pseudomonas aeruginosa, Protieus vulgaris, Klebsiella aerogenes. Previously prepared paper discs were dispensed onto the surface of the inoculated agar plate. Each disc was pressed down firmly to ensure complete contact with the agar surface. The discs were placed on the medium suitably apart and the plates were incubated at 5°C for 1h to permit good diffusion and then transferred to incubator at 37°C for 24h. After completion of 24h, the plates were inverted and placed in an incubator set to 37°C for 24h.

RESULTS: The essential oil obtained from Z.cernuum was yellow in colour. The yield is 0.007% of fresh weight. The details of compounds identified by GC-MS analysis are given in Table 1

Table 1: CHEMICAL COMPOSITION OF ESSENTIAL OIL OF Z.CERNUUM

RI*: R.P. Adams, Identification of essential oil components by Gas Chromatography/mass spectrometry 4th edition (2007) Allured Publishing  Corporation, Carol Stream, IL

RI**: Calculated by GC/MS using n-alkane series under the same conditions as for the  Sample

The major (above 10%) compounds constituting the oil are trans-Caryophyllene, α-Humulene and δ-3-Carene from Z.cernuum. These and the other compounds present in the oil from different plant sources exhibit diverse biological properties. trans-Caryophyllene and α-Humulene show anti-inflammatory properties ⁹. δ-3-Carene present in Z. cernuum is found to exhibit antimicrobial properties ¹⁰. Terpinen-4-ol was able to induce caspase-dependent apoptosis of melanoma cells and this effect was more evident in the resistant variant cell population. It also shows anti-inflammatory properties ^{11, 12, 13}. The activity of essential oil against the microorganism under study can be concluded from their respective zone of inhibition diameter which is given in Table 2.

TABLE 2: ANTIMICROBIAL SCREENING OF ESSENTIAL OILS

Standard (STD) – Ciprofloxacin 2µg/disc

Solvent – DMSO (Shows nil effect against the micro organisms under test)

It was observed that the essential oil exhibit biological activity, hampering the growth of one or the other organism. It shows susceptibility to both

gram +ve and gram -ve bacteria. Table 3 shows the minimal inhibitory concentration.

TABLE 3: MINIMAL INHIBITORY CONCENTRATION

Standard (STD) – Ciprofloxacin 2µg/disc

NI – No Inhibitory effect

 DISCUSSION: Essential oil from Z.cernuum shows good activity.  This activity can be attributed to the compounds present in the oils. The higher antimicrobial activity in essential oil may be due to the presence of major components or due to the synergistic effect of the major and minor components. Usually the major components are responsible for the antimicrobial activity of plant essential oil, but the minor components also play major role making the whole oil more active than the combination of major components in synergism.

CONCLUSION: It can be concluded that the essential oil from the rhizomes of Z.cernuum growing in Kerala have major components such as trans-Caryophyllene, α-Humulene and δ-3-Carene which were already reported for their potential biological properties. The oil also possesses considerable anti-bacterial activity against gram positive and gram negative bacteria in vitro. The component present in the oil is responsible for its antibacterial activity.                              .

ACKNOWLEDGEMENT: P.M. Shafi is thankful to UGC, New Delhi (SAP Programme) for financial assistance.

REFERENCES:

1. Kasarkar AR and Kulkarni DK: Phytochemical investigation of the genus Zingiber from the family Zingiberaceae. International Journal of pharmaceutical research and allied sciences 2011; 2:648-649.
2. Kasarkar AR and Kulkarni DK: Mineral investigation from genus Alpinia and Zingiber from south western Maharashtra. Journal of Pharmacy Research 2012; 5: 3913-3915.
3. Sanjay J: Phytochemical Screening, Antioxidant activity, Multi-Elemental and Flavonoid Analysis of Rhizome Extracts of Zingiber cernuum Dalzell. Journal of Academia and Industrial Research 2015; 4: 56-64.
4. NIST/EPA/NIH Mass Spectral Database, Version 2.1 Perkin-Elmer Instrument LLC, Copyright © 2000.
5. D Joulain, WA Konig. The atlas of spectral data of sesquiterpene hydrocarbons. E.B. Verlag, Hamburg, 1998.
6. Adams RP: Identification of Essential Oil Components by Gas Chromatography/Mass Spectrometry, 4th ed. Allured Publ. Corp., Carol Stream, IL, 2007.
7. NCCLS (National Committee for Clinical Laboratory Standards), 2000a.  Performance standards for antimicrobial disc susceptibility tests. Approved. Standard, M2-A7.
8. Bayer AW, M. Kirby DK, Sherris JC and Trick M: Antibiotic Susceptibility testing by   standard single disc diffusion method. Am J Clinical Pathol. 1966; 45: 493-496.
9. Elizabeth SF, Giselle FP, Rodrigo M et al:  Anti-inflammatory effects of compounds α-humulene and (−)-trans caryophyllene isolated from the essential oil of Cordia verbenacea. European Journal of Pharmacology 2007; 569: 228-236.
10. Cosentino S, Barra A, Pisano B et al.: Composition and antimicrobial properties of Sardinian juniperus essential oils against food borne pathogens and spoilage microorganisms. J Food Prot 2003; 66: 1288-91.
11. Annarica C, Annarita S, Laura T et al.: Terpinen-4-ol, The Main Component of Melaleuca Alternifolia (Tea Tree) Oil Inhibits the in vitro Growth of Human Melanoma Cells. Journal of Investigative Dermatology 2004; 122: 349-360.
12. Hart PH, Brand C, Carson CF et al.:  Terpinen-4-ol, the main component of the essential oil of Melaleuca alternifolia (tea tree oil), suppresses inflammatory mediator production by activated human monocytes. Inflamm Res 2000; 49: 619-626.
13. Francesca M, Flavia De B, Antonietta G et al.: In vivo activity of terpinen-4-ol, the main bioactive component of Melaleuca alternifolia Cheel (tea tree) oil against azole-susceptible and - resistant human pathogenic Candida species. BMC Infectious Diseases 2006; 6:158.
    

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How to cite this article:

Thambi M and Md Shafi P: Rhizome Essential oil Composition of Zingiber Cernuum and its anti Microbial Activity. Int J Pharm Sci Res 2016; 7(2): 829-33.doi: 10.13040/IJPSR.0975-8232.7(2).829-33.

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