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Evaluation of Antibacterial and Anticough Forming Effects of Zingiber officinale

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Article  in  Journal of Chemical and Pharmaceutical Research · December 2012

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 Pharmacology Research Article
International Journal of Chemical And Pharmaceutical Research ISSN 2319-1716
Volume 1, Issue 6, December 2012.
© Glorigin Lifesciences Private Limited.

EVALUATION OF ANTIBACTERIAL AND ANTICOUGH FORMING EFFECTS OF

Zingiber officinale EXTRACT
Wasim Raja*1, Sonam Pandey2, Sarfaraz Hanfi1 and Aafrin Khan1

Abstract
The present study was carried out to the antibacterial and anticough forming activity of well-known
herbal medicinal plant Zingiber officinale extract. The dietary intake of herbal medicine has increased in
recent years. In this experiment the antibacterial property of different concentration of plant extract was
monitored using ‘disc diffusion assay’. Antibacterial activity was screened for three microorganisms:
Proteus mirabilis, Klebsiella pneumoniae and Streptococcus aureus. The data was compared to that of
standard antibiotics. To antibacterial activity of Zingiber officinale data revealed the sensitizing quality of
extract against Proteus mirabilis, Klebsiella pneumoniae and Streptococcus aureus. 250 and 500 mg/kg
concentration of extract were having good activity, showing zone of inhibition after 12 hour time interval.
Another set of experiment anticough forming activity of Zingiber officinale extract shows the expiratory
effort due to an end tracheal mechanical stimulus was reduced by Z. officinale extract shows the dose-
response in SGOT and SGPT enzyme as compared to SO2 treated group. The mortality rate was observed
to be nil in all experimental groups. A significant reduction in body weight gain was observed. Serum
SGOT and SGPT concentration showed a significant increase as compared to control.

Author Affiliations: 1Genesis Institute of Biomedical Research, Bhopal, Madhya Pradesh India
2
Priyamvada Birla Cancer Research Institute, Satna, Madhya Pradesh, India.

Keywords: Zingiber officinale, Klebsiella pneumoniae, Proteus mirabilis, Klebsiella pneumoniae,

Streptococcus aureus, SGOT and SGPT

* Corresponding Author: Dr. Wasim Raja, Genesis Institute of Biomedical Research, Bhopal, Madhya
Pradesh, India. Email: dr.wasimraja@yahoo.com, Phone: +919425447174

Intl. J Chem Pharm Res 2012;1(6): 141-147 141 www.ijpcr.net

Wasim Raja, et al., Antibacterial and Anticough forming Effects of Zingiber officinale
1. INTRODUCTION:
Ayurveda or Ayurvedic medicine is the
most ancient of the six recognized Indian
Systems of Medicine to the Indian subcontinent.
It is sometimes considered to be a Hindu system
of health care because of its origins in the oral
advice on living from the Vedic metaphysics
(Charaka Samhita). It is used by millions of
people in India, Nepal, Sri Lanka, and
increasingly in the Western world as alternative
medicine. Ginger is carminative, pungent,
stimulant, used widely for indigestion,
stomachache, malaria and fevers. It is chiefly
used to cure diseases due to morbidity of Kapha
and Vata. Ginger with lime juice and rock salt
increases appetite and stimulates the secretion of
gastric juices. Previous animal studies had
demonstrated the anti-emetic property of fresh
ginger [1], but it was the clinical work of Mowrey
and Clayson which generated a wider interest in
[2]
this use of ginger . Some Chinese and Japanese
research found that oral and intra-gastric
application of fresh ginger decoction produced a
[1]
stimulant action on gastric secretion . Ginger
and 6-gingerol inhibited experimental gastric
[3,4]
ulcers in rats . Fresh ginger decocted in water
resulted in symptomatic improvement in 10
patients with peptic ulcers [1]. Srivastava and co-
workers found that aqueous extract of ginger
inhibited platelet aggregation induced by ADP,
epinephrine, collagen and arachidonic acid in
vitro [5]. Ginger acted by inhibiting thromboxane
synthesis [5,6]. Ginger exerted a powerful positive
inotropic effect on isolated guinea pigs left atria.
Intl. J Chem Pharm Res 2012;1(6): 141-147
Gingerols were identified as the active
[7,8]
components . The mutagenic activity of
ginger extracts has been observed in several
strains [9,10].
2. MATERIALS AND METHODS
2.1. Preparation of plant extracts
First of all ginger was cut into small
pieces. Then these small pieces of ginger were
shade dried. After drying the ginger pieces were
powdered using a mixer grinder. Then the
sample and solvent were taken in the ratio of 1:3
(sample:methanol) in Soxhlet unit. Temperature
was set around 50-60º C. the extraction was
completed in 5-6 days. After extraction, the
extract was kept in oven at 60º C for
evaporation. After evaporation extract was
available as fine crystals form. Then these
crystals were used for screening of medicinal
properties of Zingiber officinale.
2.2. Antibacterial Assay
Antibacterial activity of extracts of Z.
officinale was determined by agar disk diffusion
[3]
method at two different concentrations i.e.,
1000 mg/ml and 500mg/ml. Muller Hinton Agar
was prepared according to the manufacturer’s
instructions. The medium was sterilized by
autoclaving at 121o C for 15 minutes at 15 psi
pressure and was used for tests. Sterile molten
cool (45 o C) agar was poured aseptically into
sterile Petri dishes (15 ml each) and the plates
were allowed to solidify at room temperature in
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Wasim Raja, et al., Antibacterial and Anticough forming Effects of Zingiber officinale
sterile condition. After solidification and drying,
the plates were seeded with appropriate
microorganisms by streaking evenly on to the
surface of the medium with a sterile cotton swab
or pouring the appropriate microorganism on the
surface of dry agar plate present in peptone
broth. Care was taken for the even distribution
of culture all over the plate. The inoculums were
allowed to dry for 5 minutes. The discs of 6 mm
diameter were prepared from Whatman filter
paper No. 1 and were sterilized. The discs were
then impregnated with the extracts; solvent
DMSO and standard antibiotic i.e. Penicillin,
Gentamicin, Ampicillin and Streptomycin were
used as standard. Sterile Whatman filter paper
No 1 with different test concentrations ranging
from 1000 and 500 mg/ml/disc were placed on
to the agar with flamed forceps and gently
pressed down to ensure contact along with the
diluted extract, one appropriate control dry disc
also placed at the center. Then the plates were
incubated below 37oC for 24 hrs to allow
perfusion of drugs being tested. The next day the
zones of inhibition were measured with a
measuring scale. This experiment was carried
out in triplicate for their confirmation. The
results were read by the presence or absence of
zone of inhibition.
2.3. Anti cough forming Activity:
This activity was performed as per the
describe method of May and Widdicombe[11].
Intl. J Chem Pharm Res 2012;1(6): 141-147
2.3.1. Method of SO2 production
To 5 ml of concentrated Sulphuric acid
in glass flask, 0.5 grams of copper turnings were
added and heated in a flask on burner. The
mouth of flask was covered with aluminum foil.
After 5 minutes of heating, SO2 gas was
produced. After 5 minutes, the mouth of flask
was opened and gave the SO2 gas to the mice for
inhaling, by hanging them in front of flask
mouth for 5-10 seconds. The mice were exposed
to SO2 gas for 5-6 days. After completion of
exposure period the mice were dissected. After
dissection mice liver were flushed with normal
saline and fixed in 10% formalin. Then
histopathology study was done by the help of
histopathologist.
2.3.2. Serum biochemical analysis
The activity of SGOT and SGPT was
carried out by the method of Reitman and
Frankel[12] and expressed as IU/L. The collected
data were statistically analyzed by Student’s t-
test and the treatment groups were considered
statistically significant at P<0.05.
2.3.2.1. SGOT
Aspartate transaminase (GOT- AST)
catalyses the reaction between alpha-
ketoglutaric acid and L-aspartate giving
glutamate and oxaloacetate. Oxaloacetate, in the
presence of malate dehydrogenase (MDH) reacts
with NADH giving malate and NAD. The rate of
NADH decrease is determined photometrically
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Wasim Raja, et al., Antibacterial and Anticough forming Effects of Zingiber officinale

and is directly proportional to the GOT activity 0.9 mm (Table No.1). Thus the extract was
in the sample. showing varying activity against the
microorganism.
2.3.2.2. SGPT
The SGPT - ALT is a cellular enzyme, Table: 1 Effect of Z. officinale in Antibacterial
activity
found in highest concentration in liver and Microorganism
Test
kidney. High levels are observed in hepatic (Zone of Inhibition in mm)
Sample
Concentration (%) S. K. P.
disease like hepatitis; disease of muscle and aureus pneumonia mirabilis
traumatisms, its better application is in the Z. officinale,
7 6 9
(500 mg/ml)
diagnosis of the disease of the liver. Z. officinale,
9 7 9
When they are used in conjunction with (1000 mg/ml)
PENICILLIN- G
AST aid in the diagnosis of infarcts in the < 16 < 00 < 17
10 units/disc
myocardium, since the value of the ALT stays GENTAMICIN
< 24 < 00 < 33
within the normal limits in the presence of 120 mcg/disc
AMPICILLIN
elevated levels of AST. < 32 < 00 < 26
10 mcg/disc
STREPTOMYCIN
< 22 < 16 < 30
300 mcg/disc
3. RESULTS AND DISCUSSION
The initiation of microbial growth was
On comparing the zone of inhibition of
considered as zero hour and further accordingly
extract to that of standard antibiotics
reading was taken. Our present study showed
(Streptomycin, Gentamicin, Ampicillin and
that antibacterial activity of Zingiber officinale
Penicillin) (Table no. 1), extract showed better
extract against Streptococcus aureus which was
activity than Gentamicin & Ampicillin but
best in 1000 mg/ml concentration after 12 hrs.
extract is not potent than erythromycin &
The 0.9 mm zone of inhibition was obtained.
amoxicillin in these conditions (Table 1).
Although 500 mg/kg concentration was having
mild effect as 0.7 mm zone of inhibition. In
3.1. Cough Reflexes:
Klebsiella pneumoniae 1000 mg/ml
The expiratory effort due to an end
concentration of extract is having good
tracheal mechanical stimulus was reduced by
antibacterial activity at maximum zone of
Zingiber officinale extract shows the dose-
inhibition 0.7 mm. On the other hand 500 mg/kg
response in SGOT and SGPT enzyme as
is showing same activity, with zone of inhibition
compared to SO2 treated group. The mortality
of 0.7 mm (Table 1). For Proteus mirabilis 1000
rate was observed to be nil in all experimental
mg/ml and 500 mg/ml both concentration of
groups. A significant reduction in body weight
extract showed a maximum zone of inhibition
gain was seen. Serum SGOT and SGPT
Intl. J Chem Pharm Res 2012;1(6): 141-147 144 www.ijpcr.net
Wasim Raja, et al., Antibacterial and Anticough forming Effects of Zingiber officinale

concentration showed a significant increase as affecting the cells nearest the central vein while
compared to control (Table 2). toxic substances in the blood would show up
first in cells near the portal triad. Liver
Table: 2 Effect of Z. officinale extract on cough macrophages, called Kupffer Cells, are present
forming activity using SGOT and SGPT enzyme in sinusoids.
Body Weight
S.No Groups SGOT SGPT
Initial Final
4. CONCLUSION
Untreated 22.03 23.03 72.54 76.01±
1. The dietary intake of herbal medicine
control ± 0.62 ± 0.26 ± 0.53 o.77
has increased in recent years. In this experiment
Positive 21.56 19.86 52.85 57.11±
2. the antibacterial property of different
control ± 0.47 ± 0.69 ± 0.56 0.55
concentration of plant extract was monitored
Standard 21.77 23.03 74.12 78.23±
3. using ‘disc diffusion assay’. Antibacterial
Benadryl^ ± 0.63 ± 0.75 ± 0.85* 0.47*
Zingiber 23.16 23.06 62.47 65.49± activity was screened for three microorganisms
4.
officinale ± 0.53 ± 0.62 ± 0.51* 0.56* Proteus mirabilis, Klebsiella pneumoniae and
*denotes statistically significant as compared to Streptococcus aureus. The data was compared to
positive control group at p<0.05. that of standard antibiotics. To antibacterial
activity of Zingiber officinale data revealed the
^ Commercially available Benadryl was used as
sensitizing quality of extract against Proteus
Standard. Benadryl contains diphenhydramine as
active ingredient and is a brand name cough and mirabilis, Klebsiella pneumoniae and
allergy medicine marketed over-the-counter by Streptococcus aureus. 250 and 500 mg/kg
Johnson & Johnson subsidiary McNeil Consumer concentration of extract were having good
Healthcare. activity, showing zone of inhibition after 12
hour time interval.
3.2. Histology of Liver:
There are reports which showed
Result shows treated group histology
antibacterial activities of Zingiber officinale. As
slide of the liver the anatomical lobule with the
Proteus mirabilis, Klebsiella pneumoniae are
central vein in the center is useful for the study
gram negative and Streptococcus aureus is gram
of liver morphology, but doesn't give much
positive and our extract is having good activity
information about the functional units of the
against all three. Naturally occurring
liver. With the portal triad in the center of the
antioxidants have been extensively studied for
functional lobule and blood flowing from this
their capacity to protect organism and cells from
center in all directions toward various "central
oxidative damages. Many plant constituents
veins" some liver disease can be better
including Zingiber officinale appear to be potent
explained. For example reduced arterial blood
supply would manifest itself initially by
Intl. J Chem Pharm Res 2012;1(6): 141-147 145 www.ijpcr.net
Wasim Raja, et al., Antibacterial and Anticough forming Effects of Zingiber officinale

antioxidant property reported by other and our good alternative for broad spectrum antibiotic
system using in vitro fenton reaction. and anticough forming effect. Further studies are
Early animal studies had demonstrated needed to verify this hypothesis.
[1]
the anti-emetic property of fresh ginger , but it
was the clinical work of Mowrey and Clayson ACKNOWLEDGMENTS
which generated a wider interest in this use of The authors are thankful to Director,
[2]
Zingiber officinale . Genesis Institute of Biomedical Research,
We were also found the anti cough Bhopal, (M.P.) India for providing facility and
forming activity of Zingiber officinale extract as technical support to carry out the above work.
compared to standard (Benadryl) and control
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