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HONEY BASED POLYHERBAL ANTI TUSSIVE AND EXPECTORANT
AND THEIR ANTI-BACTERIAL ANALYSIS
Syed Asadulla*, Champa Kumari S., Fathima Fayiza, Fathimath Fayiza M., Fathima
Unaisa U.K., Fayis Muhammed K. and Mintu Aleyas
Department of Pharmacognosy, Malik Deenar College of Pharmacy, Kasaragod, Kerala.
ABSTRACT
The extracts of dried powders of Lobelia (Lobelia erinus),
Pushkaramoola root (Inula racemosa), Tulsi (Ocimum sanctum),
vasaka(Adhatoda vasika), Yastimadhu (Glycyrrhiza glabra), Long
pepper(Pipper longum) etc and also plants which enhance thier anti
microbial activity like cardamom, mustard, cinnamon, kantakari, etc
was selected in the present study for developing poly herbal syrup. The
prepared polyherbal syrup was evaluated immediately after
preparation. The purpose of this research was to formulate honey
polyherbal antitussive and expectorant and to evaluate antimicrobial
activity of these preparation againt S.aureus and E.coli by using agar
well diffusion method using different concentration of extracts.
KEYWORDS: Polyherbal, formulation, syrup, liquid dosage form, S.aureus, E.coli.
INTRODUCTION
There are many types of drugs that are used to suppress cough and are often prescribed in
combination. Before dealing with the particular type of drug used, it is important to consider
briefly the nature of cough production, its role in disease and desirability of suppressing it.
The studies showing that about 95 million units of pediatric drugs were sold for cough and
cold every year in India. Commercially available cough syrups containing synthetic and
semisynthetic have several disadvantages. To overcome such disadvantages naturally
occurring herbs having antitussive and expectorant activity can be used individually or in
combination. The herbal medicines are normally considered safer than the non -herbal
medicines because the naturally active ingredients in the herbal medicines are in combination
with other component.
WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES
SJIF Impact Factor 7.632
Volume 9, Issue 11, 1768-1779 Research Article ISSN 2278 – 4357
*Corresponding Author
Syed Asadulla
Department of
Pharmacognosy, Malik
Deenar College of
Pharmacy, Kasaragod,
Kerala.
Article Received on
06 Sept. 2020,
Revised on 27 Sept. 2020,
Accepted on 18 October 2020
DOI: 10.20959/wjpps202011-17686
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Cough syrups are the medicated liquid preparations used for relieve coughing. Cough syrups
can be prepared by using traditional drugs. The herbal cough syrups are available in the
market and Are more effective against cough. In the recent times the use of herbal cough
syrups is on the rise due to the spread of the awareness of the effect of complementary and
alternative medicine. It is also due to the much stronger believe that alternative therapy is
with less side effects, that makes the demand of herbal products in the market. People are
willing to follow alternative therapy since nowadays they are well bothered about the side
effects of organic medicines.
Herbal cough syrups are prepared by using honey as vehicle. The stability and fragrance of
the preparation can be improved by addition of excepients. Herbal formulations were
generally examined through physicochemical as well as pharmacological studies. pH,
viscosity, skin irritations, diffusion studies, visual appearance and stability studies.
Combination of plant drug extract shows synergistic activities like antimicrobial activity.
METHODOLOGY
Plant Collection
The plants were collected from Bangalore. The plant material was taxonomically identified
by the botanist Mr. Dr. V S Anil Kumar, H O D, department of botany, Govt college
Kasaragod. The plant parts were dried and then powdered with mechanical grinder and stored
in an air tight container.
PREPARATION OF EXTRACT
Maceration
Weighed about 25 g of powdered crude drug, to this added 100 ml of water and 10 g of
sucrose(10% of water taken) and boiled for 2 hrs. Allowed to cool to this added 1.5 g of yeast
and kept for 7 days. Then the mixture were filtered through muslin cloth and evaporated to
get the extract
Preparation of Artificial Honey
For the preparation of honey first we have taken the microorganism Aspergillus niger. Which
is obtained from the bulbs of Onion as well as garlic. It can be seen in the decade bulbs in the
form of black powder. It is then collected and cultured using Sabouraud’s dextrose agar
medium. The fully grown fungi were obtained after 5 days.
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In this honey we are giving a pineapple flavor so that the palatability can be enhanced. For
that we have taken a large pineapple, it is then cut in to small pieces, blended it and allowed
to boil for the separation of fibers about 2-3 hrs. Then the juice was filtered using muslin
cloth.
For the artificial honey we have to prepare immobilized Aspergillus beads. This beads can be
prepared by following procedure, 2% of sodium alginate solution was prepared by dispersing
and mixing 1 g in 50 ml of Tris buffer HCl. The Aspergillus separated from culture medium
was added to 50 ml of alginate solution and mixed thoroughly to make dispersion. The gel
was then taken into a syringe and added drop wise to calcium chloride solution of volume 50
ml. It was then kept for 20 minutes and beads obtained were washed with dist. Water and
then with buffer. It was then dried to room temperature.
Here we tried to prepare the best honey for that we have taken different concentration of
pineapple juice as well as sucrose using 6 beakers. Each beaker was added with 100 beads of
Aspergillus. To this 90 ml, 85 ml, 80 ml, 75 ml, 70 ml, 66.4 ml pine apple juice was added
from 1-6 beakers. Similarly sucrose was added in concentration of 10%, 15%, 20%, 25%,
30%,33.6% respectively. Then the beakers were kept for 7 days and observed regularly.
The best honey was obtained in the 6 th beaker which contained high amount of fructose. It
was then taken and filtered through muslin cloth for the separation of beads. Then it was
warmed and allowed to cool. Now the artificial honey is ready to use.
1) 90 ml juice + 10% sucrose 2) 85 ml juice + 15% sucrose
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3)80 ml juice + 20% sucrose 4) 75 ml juice +25% sucrose
(5) 70 ml juice + 30% sucrose (6) 66.6 ml juice+ 33.4% sucrose
PREPARATION OF POLYHERBAL ANTITUSSIVE AND EXPECTORANT
Formulation For Antitussive
Aniseed 10mg
Thyme 10mg
Peppermint 30mg
Cinnamon 15mg
Pushkaramoola root .................................. 250mg
Kantakari plant 150mg
Ocimum sanctum ...................................... 250mg
Vasaka leaf 250mg
Yeshtimadhu stem ..................................... 250mg
Shunti root 250mg
Pippal root 250mg
Pineapple flavoured honey .......................................... qs
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Formulation For Expectorant
Lobelia 50mg
Liquorice 100mg
Wild cherry 15mg
Black mustard seed…… 15mg
Black pepper 30mg
Cardamom 30mg
Ammonium chloride ............................... 0.138g
sod. citrate 57.03mg
Menthol 1.14mg
Nilgiri oil 1.2mg
Pineapple flavoured honey ............................. qs
Procedure
Sufficient quantity of drug extract were weighed and dissolved in small amount of prepared
honey. These are mixed well and add preservative and other additives in remaining amount of
honey. And mix the two solutions. Volume was made to 10 ml with honey.
Physico Chemical Evaluation of Anti Tussive And Expectorant
The physical parameters of antitussives and expectorants like colour, appearance, pH were
evaluated.
Visual Appearance
The preparations were inspected for its clarity, color and transparency. They were also
evaluated for the presence of particles.
Stability Study
The purpose of stability testing is to provide evidence on how the quality of drug substance
varies with time, under the influence of variety of factors such as temperature, humidity and
light. And to establish a retest period for the drug substance or a shelf life for the drug
product and recommended storage conditions.
Placebo and cough syrup were evaluated for the thermo stability, 10 ml of the antitussive an
10 ml of expectorant was taken in 25 ml beaker, each were taken in 2 beakers, one was kept
at room temperature and the other was kept at 40˚C for 2 weeks. It was also examined for pH,
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appearance and homogenecity.
p H STUDIES
Well calibrated pH meter was used to determine the pH of cough syrups. The result were
documented and compared over 2 weeks.
Antibacterial Activity Test
Individual plant extracts and formulated antitussive and expectorant are studied for its anti-
microbial properties. However there are no reports regarding the evaluation of antimicrobial
activity of these herbs in combination. Hence the drug was evaluated for antimicrobial
activity in S. aureus and E. coli
Screening of Antibacterial Activity
Table No 2: Screening Of Antibacterial Activity Preparation Of Culture Media
Preparation Of Nutrient Agar Media.
ORGANISM S. aureus, E. coli
METHOD Agar well diffusion method
MEDIA Nutrient agar media
STANDARD Ciprofloxacin
TEST Plant extracts and formulations
Suspended 1.4 g in 50 ml dist. Water, heat to boiling to dissolve the medium completely.
Sterilized by autoclaving at 15 Ibs pressure. 121ºC for 15 minutes
Screening By Agar Well Diffusion Method
Nutrient agar was prepared. Then it was poured on sterilized petri plate and allowed to
solidify. The bacterial culture was swabbed over the plates. Then, the wells were prepared by
using sterilized cork borer. To that the separate drug extract and combined drug extract as
well as std ciprofloxacin was added. Then the plates were incubated at 37˚C for 24-48 hrs.
The zone of inhibition was measured in mm.
Confirmation of Fructose Content In Honey
For the confirmation of fructose content in honey seliwanoff’s test and Osazone test was done
Seliwanoff’s Test
To 3 ml of seliwanoff’s reagent, add 1 ml of the test solution. Boil in water bath for 2
minutes. A cherry red colour precipitate with in 5 minutes was obtained.
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Osazone Test
To 5 ml of test solution add 10 drops of glacial acetic acid 1 g of crystalline sodium acetate,
0.5 g of phenyl hydrazine hydrochloride and mix well. Keep the tube boiling for 20 minutes
and cool and observed under microscope.
RESULT AND DISCUSSION
Plant Collection and Drying
Dried plant parts used for the study was collected from Bangalore, Karnataka and some from
Kasaragod too. The drugs were identified and authenticated by Dr. V.S Anil kumar, HOD,
Govt. college, Kasaragod.
Preparation of Extract
The extract were prepared by maceration. The extract were evaporated. The percentage yield
of extracts were calculated and result are shown below
Drugs % Yield of Extracts
Lobelia 13.8%
Kantakari 9%
Aniseed 9.7%
Black mustard 8.5%
Black pepper 7%
Vasaka 10%
Tulsi 8.7%
Pepper mint 10.2%
Liquorice 12.5%
Pushkaramoola 11%
Ginger 6.8%
Thyme 6.6%
Wild cherry 10.8%
Cinnamon 11.4%
Cardamom 10%
Long pepper 9%
Formulation of Polyherbal Anti Tussive And Expectorant
The poly herbal antitussive and expectorant were prepared on the basis of the formula given
Visual Appearance
When the optimized formula was evaluated for 4 weeks the visual appearance of the
formulations were found to be unchanged, stable and acceptable.
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Stability Study
The formulations were stored under and observed under room temperature and elevated
temperature (40˚C) for 4 weeks. On observation the formulations were found to be stable,
unchanged and good in appearance. Hence it was found to be thermo stable.
Table: stability study.
Formulation Appearance Homogenicity Ph
Anti tussive Stable and
unchanged Maintained 4
Expectorant Stable and unchanged Maintained 4.2
Evaluation of Antibacterial Activity
Table: evaluation of antibacterial activity.
Sample Concentration(µl)
Zone of inhibition(mm)
Bacterial culture
S. aureus E. coli
Lobelia
50 10 mm 11 mm
75 12 mm 13mm
100 13 mm 15mm
Liquorice
50 9 mm 12 mm
75 10mm 12mm
100 11 mm 13 mm
Kantakari
50 10 mm 10 mm
75 11 mm 12 mm
100 12 mm 13 mm
Ginger
50 10 mm 11 mm
75 12 mm 13mm
100 13 mm 14 mm
Cardamom
50 8 mm 10 mm
75 11mm 10 mm
100 10 mm 13 mm
Pushkaramoola
50 11mm 14 mm
75 12mm 15mm
100 14mm 15 mm
Vasaka
50 9mm 10mm
75 11 mm 12mm
100 11mm 13mm
Pepper mint
50 +9mm 10mm
75 11mm 12mm
100 13mm 14mm
Black pepper
50 8mm 9mm
75 10mm 11mm
100 11mm 12mm
Black mustard 50 10mm 12mm
75 11mm 13mm
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100 11mm 14mm
Long pepper
50 9mm 10mm
75 10mm 12mm
100 9mm 10mm
Aniseed
50 10mm 13mm
75 11mm 14mm
100 14mm 15mm
Thyme
50 8mm 9mm
75 10mm 10mm
100 11mm 13mm
Tulasi
50 10mm 12mm
75 13mm 14mm
100 14mm 15mm
Wild cherry
50 7mm 8mm
75 9mm 9mm
100 10mm 11mm
Cinnamon
50 10mm 12mm
75 11mm 13mm
100 12mm 14mm
Anti tussive 100 15 mm 19 mm
Expectorant 100 18 mm 21 mm
Standard Ciprofloxacin 20mm 25 mm
Pushkaramoola Aniseed Mustard Thulasi Ginger
Cardomom Kantakari Thyme Liquorice Black pepper
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Lobelia Pepper mint Vasaka Wild cherry Long pepper
Cinnamon
Anti Bacterial Effect of Formulation
A- S. aureus B-E,coli C- S. aureus D-E. coli
A – Antibacterial activity of antitussive against S. aureus
Antibacterial activity of antitussive against E. coli
C - Antibacterial activity of Expectorant against S. aureus
D - Antibacterial activity of Expectorant against E. coli
FORMULATIONS
Pineapple honey Expectorant Antitussive
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Identification of Fructose Content In Honey
By molisch’s and fehling’s test it was found that the prepared pineapple flavored honey
contains fructose. For the confirmation also done the osazone test and the seliwanoff’s test.
The result of osazone test is given below
Cherry red Broom stick
CONCLUSION
The present study aimed at the formulation of Honey based polyherbal anti tussive and
expectorant and their Anti- bacterial analysis
Extraction of various crude drugs were done by Soxhlet extraction technique (methanolic
extract), and maceration technique (aq.extract).
Polyherbal antitussive and expectorant were prepared using honey as the solvent, menthol,
thymol, Ammonium chloride, it yielded a formulation of satisfactory phytochemical
appearance.
The stability study of the formulation was carried out.
Studied the Anti-bacterial activity of the individual extract as well as the formulation against
S. aureus and E. coli.
As per our research work, it reveals that the drug extracts have a good Antibacterial activity,
How ever further studies to be done to improve the stability of the formulations.
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