FORMULATION AND EVALUATION OF

ANTI-BACTERIAL HERBAL OINTMENT

Project submitted to
JAWAHARLAL NEHRU TECHNOLOGICAL
UNIVERSITY ,HYERABAD
ASFIYA UNNISA - 15VH1R0007
AYESHA SHIREEN - 15VH1R0009
BUSHRA FATIMA - 15VH1R0010
IMRANA BEGUM - 15VH1R0018
SANA BEGUM - 15VH1R0030

Under the guidance and supervision of

MR. PAJJURI VENKANNA.
M.Pharm
Dept of Pharmacognosy

MAK COLLEGE OF PHARMACY,

MOINABAD,R.R DIST, HYDERABAD,
TELANGANA
 Overview
• Introduction
• Aims and Objective
• Plan of work
• Plant profile
• Materials and methods
• Phytochemical analysis
• Formulation of ointment
• Evaluation of ointment
• In vitro anti- microbial studies
• Results
• Conclusion
• Refrences
 INTRODUCTION
• Medicinal plants, also called medicinal herbs, have
been discovered and used in traditional medicine
practices since prehistoric times.
• Plant drugs are frequently considered to be less toxic and
freer from side effects than the synthetic ones.
• Along with other dosage forms, herbal drugs are also
formulated in the form of ointment.
 INTRODUCTION (contd…)
DESIRABLE PROPERTIES OF OINTMENT:
• Stable under normal conditions of use and storage.
• Non-reactive and compatible.
• Free from objectionable odor.
• Non toxic.
• Non sensitizing.
• Aesthetically appealing.
• Easy to apply.
• Non greasy.
• Non irritating.
 INTRODUCTION( contd…)
TYPES OF OINTMENT BASES:
There are 5 classes or types of ointment bases:
• Oleaginous bases.
• Absorption bases.
• Water in oil emulsion bases.
• Oil in water emulsion bases.
• Water soluble or water
miscible bases.
 INTRODUCTION(Contd…)
• Trachyspermum ammi is a small erect, annual,
herbaceous plant with branched leafy stems, feather
like leaves.
• The fruits are minute, greyish brown colored and egg
shaped.
• Seeds contain an essential oil containing about 50%
thymol which is strong germicide, anti-spasmodic
and fungicide
 AIMS AND OBJECTIVE
• To evaluate the anti-bacterial activity of herbal
formulation of the collected plant seed i.e.,
Trachyspermum ammi (Ajwain).
 PLAN OF WORK
SELECTION OF PLANT AND LITERATURE REVIEW.

COLLECTION OF SEEDS

CONVERSION OF SEEDS INTO POWDER

MACROSCOPIC AND MICROSCOPIC CHARACTERS

SELECTION OF SUITABLE METHODS OF EXTRACTION

FORMULATION AND EVALUATION OF OINTMENT

EVALUATION OF ANTI MICROBIAL ACTVITY

 PLANT PROFILE
• Common name: Ajwain
• Botanical name: Trachyspermum Ammi
• Family: Apiaceae
• Venicular names:
Sanskrit: Yamini
Assamese: Jain
Bengali: Yamini
English: Bishop’s weed
Telugu: Vamu
 PLANT PROFILE(Contd…)
Taxonomical classification:
• Kingdom: Plantae, Plant
• Subkingdom: Tracheobionta, Vascular plant
• Superdivision: Spermatophyta, Seed Plant
• Division: Magnoliophyta, flowering plant
• Class: Magnoliopsida, Dicotyledons
• Order: Apiales
• Family: Apiaceae
• Genus: Trachyspermum
• Species: Ammi
 PLANT PROFILE(Contd…)
Botanical Description:
 Climate
 Soil
 Preparation of land
 Sowing time
 Seed rate
 Seed treatment
 Sowing method
 Manures and fertilizers
 Irrigation
 Weed management
 Plant protection
 PLANT PROFILE(Contd…)
MACROSCOPIC DESCRIPTION:
• Ajowan is an erect, glabrous or minutely pubescent,
branched spice herb which reaches up to 90 cm in height.
Stem is straight, much branched and usually puberulous.
• Flowers are terminal or seemingly-lateral pedunculate,
compound umbels, white, small, pedicels (0.5-4mm) and
unequal. Calax teeth are conspicuous, minute, ovate or
obsolete.
 PLANT PROFILE(Contd…)
MICROSCOPIC DESCRIPTION:
• The transverse section of fruit shows two hexagonal
structures attached with each other by carpophores
• Epicarps consists of a single layer of tangentially
elongated tabular cells it also consist of mesocarp
which has thick-walled, rectangular elongated cells
having some vittae.
 PLANT
PROFILE(Cont…)
MEDICINAL USES:
• Ajwain is used as germicide and antiseptic, it is
used in cough syrups,throat lozenges, asthma,
indigestion etc.
• Used in treatment of abdominal tumor,
enlargement of spleen, piles, vomiting etc.
• Useful in alleviating spasmodic pains of stomach
and intestine.
• It is a good digestive.
• Relieves heart ache when taken with hot water.
 PLANT PROFILE(Cont…)
HERBAL MEDICINAL USES:
• In pregnancy
• Venomous insect bite
• Toothache and headache
• Hiccups
• Kidney stone
• Stomach tonic
• Acidity
 MATERIALS AND METHODS
• Collection of plant material.
• Preparation of plant extract.
• Solvent extraction using soxhlet apparatus.
Materials required:
 Ajwain seeds
 Soxhlet apparatus
 Heating mantle
 Methanol
 Measuring cylinder
 Petri plates etc
 PHYTOCHEMICAL
SCREENING
S no Constituents Methanolic extract
1 Alkaloids +
2 Proteins +
3 Carbohydrates +
4 Oils &fats +
5 Resins +
6 Phenols +
7 Starch +
8 Tannins -
9 Steroids -
10 Terpenoids -
 FORMULATION OF
OINTMENT
Working formula for preparation of ointment:
S.NO COMPONENTS F-1 F-2

1. Methanolic Extract 2 4

2. Emulsifyning wax 14 14

3. White soft paraffin 25 25

4. Liquid paraffin 10 10
 FORMULATION OF
OINTMENT(contd…)
Methadology:
Fusion method
• Fusion method is the act or procedure of
liquefying or melting by the application of heat.
By the fusion method all the components of the
ointment are combined by melted together and
cooled with constant stirring until congealed.
EVALUATION OF FORMULATED
OINTMENT
Physicochemical parameters:
• Color and odor.
• Loss on drying.
• Diffusion study and
• Irritancy test
• Penetration
 IN VITRO ANTI-MICROBIAL
STUDIES
(1) Preparation of Agar (2) Laminar air flow unit

(3)Preparation of Agar plates (4) Inoculation of Agar plates.

 RESULTS
Literature revealed that the selected plant that
Trachyspermum ammi (seeds) revealed antibacterial
activity.
Extraction and phytochemical screening was done using
methanol as solvent. Phytochemical screening
confirmed the presence of various phytoconstituents
like oils and fats, alkaloids, proteins, carbohydrates,
resins, phenols, starch etc.
 Physiochemical constituents of
formulated ointment
S no Phytochemical Formulation- 1(2%) Formulation- 2(4%)
parameters

1 Colour Yellowish green Dark yellowish green

2 Odour Characteristic Characteristic

3 Loss on drying 0.40 0.51

5 Diffusion study 0.2cm 0.3cm

6 Spreadability Good Good

 Antibacterial activity of formulated
ointment showing zone of inhibition:

S no Organisms Formulation 1(2%) Formulation 2 (4%) Standard

1 E.coli 6.5 7.8 8.0

2 P. aeroginosa 7.1 8.3 8.9

3 S. aureus 8.4 9.1 10.7

Zone of inhibition against the
bacteria
 CONCLUSION
In the present study the methanolic extract of Trachyspermum ammi
against the test organisms brought the suggestion that the above
plant have antibacterial activity, tested against the micro-organisms
that are E.coli, P.aeroginosa( gram negative bacteria), S.aureus(
gram positive bacteria) in concentrations of 2%,4% in the form of
ointment prepared by fusion method. Since the methanolic crude
extract and formulated ointment of the plant(seeds) showed the
presence of alkaloids, carbohydrates, oils & fats, glycosides, resins
content. Methanolic extract of Trachyspermum ammi showed
greater zone of inhibition towards gram-positive i.e S.aureus
(9.1mm) bacteria compare to gram-negative bacteria.
Thus, the procedure can be adopted to formulate good herbal ointment
which may lead to a new formulation for eradicating the microbial
related diseases for serving the human society.
 REFRENCES
• Dr. Mahtab Alam Khan, Introduction And Importance Of
Medicinal Plants, article 2016.
• Kamal Jeet, Nisha Devi et al., Trachyspermum ammi (ajwain):
A comprehensive review,International Research Journal of
Pharmacy 2012, 3(5).
• Saxena and Vyas, “A comprehensive review on Ajwain
(Trachyspermum ammi)”, The National Centre for
Biotechnology Information (NCBI), Pharmacognosy Review.,
1986.
• Vikas Shrivastav et al., “Anti microbial potential of Ajwain
(Trachyspermum ammi)”, Journal of Pharmacy
Research.,2012.