Journal of Pharmacognosy and Phytochemistry 2014; 3(3): 130-133

  

E: ISSN 2278-4136
P: ISSN 2349-8234 Chemical composition and biological activity of the
JPP 2014; 3(3): 130-133
Received: 08-07-2014 essential oil of rhizome of Zingiber zerumbet (L.) Smith
Accepted: 30-08-2014
Chingakham B Singh, Saikhom B Chanu, Lenin Kh, Ningombam
Chingakham B Singh
Institute of bioresources and
Swapana, Charles Cantrell, Samir A Ross
sustainable development, Imphal-
795001, Manipur, India, Abstract
National center for natural The aim was designed to study the biological activity and chemical composition of essential oil of
products research, University of Zingiber zerumbet (L.) Smith. The essential oil extracted from the rhizome of the plant was analysed by
Mississippi, University, MS gas chromatography-mass spectroscopy and its major components amounting to 98.4 % were found to be
38677, USA. zerumbone (75.2%), α- caryophyllene (7.1%), camphene (5.1%), eucalyptol (2.4%), and camphor
(3.0%). Antioxidant activity and total phenol content assay were studied using the DPPH and Folin-
Saikhom B Chanu Ciocalteu colorimetric methods. Antimalarial, antileishmanial, antimicrobial assays were analysed for the
Institute of bioresources and essential oil and its major component, zerumbone. The essential oil and zerumbone exhibited antimalarial
sustainable development, Imphal- and antileishmanial activities, whereas only Cryptococcus neoformans showed antimicrobial activity of
795001, Manipur, India. the essential oil.
Lenin Kh Keywords: Zingiber zerumbet, Essential oil, Zerumbone, Antimalarial, Antileishmanial, Antimicrobial,
Institute of bioresources and
Antioxidant
sustainable development, Imphal-
795001, Manipur, India.
1. Introduction
Ningombam Swapana Zingiber zerumbet (L.) Smith is a monocotyledonous perennial medicinal plant belonging to
Department of chemistry, s. kula the Zingiberaceae family. It is known as shampoo ginger or pinecone ginger as its rhizomes
womens College, nambol-795134, produce foaming properties likes that of shampoo. It has many different local names
Manipur, India depending on their area of vegetation and location. It grows in subtropical climates such as
Charles Cantrell
India, South-East Asian countries, South Pacific Islands and Okinawan Islands, and has been
Natural products utilization used for local folk medicine and gardening. It is called Shinkha in Manipur, North-East India.
research unit, usda-ars, university, Traditionally, it is used for the treatment of stomach ache, toothache, fever, and indigestion [1].
ms 38677, USA. It has been also used as a spice and for ulcerative colitis [2]. The essential oils of Z. zerumbet
have been studied by different authors since 1944. The major component of the rhizome’s oil
Samir A Ross varies from 12.7-73.1% [3, 4, 5, 6]. Zerumbone and α-caryophyllene has been reported as major
National Center for Natural
constituents in rhizome and leaf oils [7]. Many authors have reported different bioactivities
Products Research and
Department of Biomolecular
from the essential oils and extracts of Z. zerumbet such as anti-inflammatory [8, 9], antitumor [10,
2]
Sciences, School of pharmacy, , free radical scavenging [11], colon and skin cancer [12], antialzheimer diseases and dementia
University of Mississippi, activities [13, 14].
University, MS 38677, USA. In the present work we aimed to evaluate the antileishmanial, antimalarial, antimicrobial and
chemical composition of the essential oil. In this work we are reporting for the first time, the
antileishmanial and antimalarial activities of essential oil from Z. zerumbet and the major
compound zerumbone isolated from its rhizome essential oil.

2. Materials and Methods

2.1. Plant material
The plant materials were collected from Manipur, North-East India, 920 m from sea level,
longitude 93°58´´and latitude 24°44´´ in March, 2012. The plant was identified by the
taxonomist of the institute and accession number as IBSD/Z-42-23.

Correspondence: 2.2. Solvents and chemicals

Samir A Ross Gallic acid, ascorbic acid, 1,1-diphenyl-2-picrylhydrazyl (DPPH), ciprofloxacin, amphotericin-
Department of pharmacognosy, B chloroquine, pentamidine were obtained from Sigma Aldrich. Analytical grade solvents
school of pharmacy, University were used during the experiments obtained from the Merck chemicals, Mumbai, India and
of Mississippi, University, MS Sigma-Aldrich.
38677, USA.
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2.3. Extraction of essential oil scavenge the 50% DPPH free radical.
Fresh rhizomes were collected and washed thoroughly with
tap water. These were cut into 5-6 mm slices and put into the 2.8 Biological activity
Clevenger type oil extractor. The oil was dried over 2.8.1. Antimicrobial assay
anhydrous sodium sulphate and stored at 4±2 °C. The oil and the isolated compound, zerumbone were tested
for antibacterial activity against Staphylococcus aureus
2.4. Gas chromatography-mass spectrometry (GC-MS) ATCC 29213, methicillin resistant S. aureus ATCC
The oil was analysed by GC-MS on a Varian CP-3800 GC 33591(MRS), Escherichia coli ATCC 35218, Pseudomonas
coupled to a Varian Saturn 2000 MS/MS. The GC was aeruginosa ATCC27853, Mycobacterium intracellulare
equipped with a DB-5 fused silica capillary column (30 m x ATCC 23068 and antifungal activity against Candida
0.25 mm, with film thickness of 0.25 µm) operated using the albicans ATCC 90028, C. glabrata ATCC 90030, C. krusei
following conditions: injector temperature, 240 °C, column ATCC 6258, Cryptococcus neoformans ATCC 90113,
temperature, 60-240 °C at 3 °C per minute, then held at 240 Aspergillus fumigatus ATCC 204305. Ciprofloxacin and
°C for 5 min; carrier gas, He; injection volume, 1 µl Amphotericin-B were used as positive control for bacteria
(splitless). The MS mass ranged from 40 to 650 m/z, filament and fungi respectively [17].
delay of 3 min, target TIC of 20,000, a prescan ionization
time of 100 µ sec, an ion trap temperature of 150 °C, 2.8.2. Antimalarial activity
manifold temperature of 60 °C, and a transfer line In vitro antimalarial activity was determined against
temperature of 170 °C. chloroquine sensitive (D6, Sierra Leone) and resistant (W2,
Indo China) strains of P. falciparum by measuring
2.5. Identification of components plasmodial LDH activity. Chloroquine was used as positive
The constituents of the oil were identified using retention control [18].
times, kovats indices and mass spectra. Confirmed integrated
peaks were then used for the percentage of each chemical 2.8.3. Antileishmanial activity
constituents present in the essential oil. Kovats indices were The antileishmanial activity was tested against Leishmania
calculated using the equation: donovani promastigotes; pentamidine and Amphotericin-B
were used as positive controls [19].
KI (x) =100 [(log RT (x)-log Pz)/(log RT(Pz+1)-log RT (Pz)]
Where: RT (Pz) ≤ RT(x) ≤ RT(Pz+1), and P4…..P25 are n 2.8.4 Statistical analysis
paraffins. All IC50 values were calculated using the XL Fit curve fitting
software.
2.6. Determination of total phenolic content
The total phenolic content of the essential oil was determined 3. Results and Discussion
by the Folin–Ciocalteu colorimetric method [15]. An aliquot 3.1. Extraction yield and GC-MS analyses
(0.125 ml) of a diluted acetone sample was added to 0.5 ml The yield of essential oil was 0.12%. GC-MS analyses of the
of deionized water and 0.125 ml of the Folin-Ciocalteu essential oil led to the identification of ten major compounds
reagent. The mixture was shaken and allowed to stand for 6 accounting for the 98.4% of the oil (Table 1). Zerumbone
min, before adding 1.25 ml of 7% sodium carbonate solution. (75.2%), α-caryophyllene (7.1%), camphene (5.1%),
The solution was adjusted with deionized water to a final eucalyptol (2.4%), and camphor (3.0%) were the major
volume of 3 ml and mixed thoroughly. After incubation for components of the oil. Zerumbone was isolated in pure form
90 min at 23 °C, the absorbance versus prepared blank was and its structure was confirmed by 1H-NMR, 13C-NMR,
read at 725 nm with a UV-Visible spectrophotometer DEPT, HR-ESIMS and comparison with literature data.
(Multiskan spectrum, Thermo Scientific). Total phenolic
content was expressed as milligrams of gallic acid 3.2. Antioxidant activity and total phenolic content
equivalents per 100 gram of essential oil through the The DPPH free radical scavenging activity together with
calibration curve with gallic acid. Folin-Ciocalteu colorimetric assay was performed on the
essential oil [15]. The results were given in table 2. The
2.7. Antioxidant activity assay antioxidant activity in the DPPH radical scavenging test is
The DPPH free radical scavenging was assessed of the due to the hydrogen donating ability of the test material. The
essential oil [16]. 0.1 mM DPPH radical solution in ethanol capability of substances to donate hydrogen to convert DPPH
was prepared. After 30 min, the absorbance was measured at into the non-radical form of DPPH can be followed
517 nm on UV-Visible spectrometer (Multiskan spectrum, spectrophotometrically. The oil exhibited DPPH scavenging
Thermo Scientific). Decreasing the absorbance of the DPPH activity (IC50 1.60 µg/ml) which is comparable with the
solution indicates an increase in DPPH radical scavenging standard ascorbic acid (IC50 1.59µg/ml). The high scavenging
activity. The radical scavenging activity, expressed as activity was attributed to the high content of zerumbone (IC50
percentage of inhibition was calculated using the equation: 2.55 µg/ml). Total phenolic content of the oil was 5.05
mg/100 g and it doesn’t play much role in the high
Inhibition Concentration % = [(A-B)/A] x 100 scavenging activity of the oils [16].
Although not much data on antioxidant activity of Z.
Where A is the absorbance of control (DPPH solution zerumbet is available, solvent extracts (ethanol, methanol and
without the sample), B is the absorbance of DPPH solution in isopropanol) of Z. zerumbet spent have shown fair DPPH
the presence of the sample. scavenging activity [20] which is similar with what we are
IC50 value is the concentration of the sample required to reporting. However, other authors reported low DPPH
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scavenging activity which may be due to difference in environmental factors [21].
extraction methods, cultivars, maturity and other

Table 1: Chemical composition of Zingiber zerumbet essential oil

Compounds Retention time Kovat Index % Components

Cyclone 6.050 927.04 0.11
Alpha pinene 6.338 938.31 0.59
Camphene 6.744 953.35 5.1
Eucalyptol 9.250 1037.09 2.4
Camphor 13.506 1150.84 3.0
Humulene 26.455 1458.66 1.2
Caryophyllene Oxide 31.514 1582.35 1.91
2,6,6-trimethylundecan-1,3-dien-9-yn-5-one 32.122 1595.85 1.96
α-caryophyllene 32.526 1606.61 7.1
Zerumbone 37.209 1740.71 75.2

Table 2: Antioxidant and Total phenol content assay of Zingiber zerumbet essential oil and zerumbone

Samples IC50 (µg/ml) TPC (mg/100 g)

Rhizome oil 1.60 5.05
Ascorbic acid 1.59 ND
Zerumbone isolated 2.55 ND

ND-not determined

3.3. Antimicrobial activity 3.4. Antimalarial activity

Antimicrobial study was done for both the isolated Antimalarial activity was studied both for the oil and the
compounds and the essential oil against five fungi and five compound zerumbone against chloroquine sensitive
bacteria. The oil was found to have good activity against Plasmodium falciparum (D6, Sierra Leone) and resistant
Cryptococcus neoformans ATCC 90113 with IC50 value of (W2, Indo China). The oil showed moderate antimalarial
8µg/ml. However, no antimicrobial activity was shown activity (IC50 17.5 µg/ml and 20.0 µg/ml against P.
against zerumbone (Table 3) [17]. falciparum D6 and P. falciparum W2). Zerumbone showed
The essential oil of Z. zerumbet exhibited a negative good activity with IC50 values of 3.9 µg/ml and 4.4 µg/ml
antifungal effect using broth microdilution and disc gel against P. falciparum D6 and P. falciparum W2 (Table 3)
diffusion methods [22]. The antifungal activity was assessed [18]
.
against five dermatophytes (Trichophyton mentagrophytes, T.
rubrum, Microsporum canis, M. nanum and Epidermophyton 3.5. Antileishmanial activity
floccosum), three filamentous fungi (Aspergillus niger, A. Antileishmanial evaluation was done on the oil and the
fumigatus and Mucor sps.) and five strains of yeast compound zerumbone against Leishmania donovani. The oil
(Saccharomyces cerevisiae, C. neoformans, Candida showed moderate activity with IC50 and IC90 values of 4.6
albicans, C. tropicalis and Torulopsis glabrata). This report µg/ml and 18.0 µg/ml. Zerumbone showed good activity with
is similar with what we are reporting, however we found the IC50 and IC90 <1.6 µg/ml and 4.7 µg/ml [19].
essential oil Z. zerumbet with IC50 value of 8µg/ml against C.
neoformans.

Table 3: Biological activity of essential oil of Zingiber zerumbet and zerumbone

Biological activity Test parasite Rhizome essential oil (µg/ml) Zerumbone (µg/ml)
L. donovani IC50 4.62 <1.6
Antileishmanial activity
IC90 18.00 4.69
Antimalarial activity P. falciparum D6 IC50 17.47 3.94
W2 IC50 20.03 4.44
Antimicrobial C .neoformans 8.00 No activity

4. Conclusion its antimalarial and antileishmanial properties is on its way

In conclusion, the biological activity of the essential oil of Z. which may lead to further information about the properties of
zerumbet is attributed to zerumbone. To the best of our zerumbone.
knowledge, this is the first study of antimalarial and
antileishmanial properties of zerumbone and its rhizome 5. Acknowledgments
essential oil. A further continuation of this research work for CBS is thankful to DBT, Govt. of India for DBT Overseas
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fellowship while WRD to DBT, Govt. of India for RA, neurotrophic factors. Yakugaku Zasshi 2006; 126:747-
National Programme. SBC also thanks to DBT, Govt. of 755.
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