Project Presentation

GUIDED BY:
DR.PADMALATHA
DEPT. OF BIOTECHNOLOGY
TOPIC: COMPARATIVE STUDY OF
PHARMACOLOGICAL GROUP MEMBERS: ADMISSION NUMBER:

ASSESSMENT ON -JAPHIA RIANA

-ISHWARYA
121221572001
121221572002
SATINWOOD,MANGO, TULSI AND -SRUJAN
-VARSHA
121221572012
121221572026
INDIGO:
 Introduction
Pharmacological assessment of medicinal plants is important because it helps to understand the
therapeutic mechanisms of these plants, which can be used to treat complex diseases. Pharmacological
studies also help to identify, validate, and standardize medicinal plants.
Ocimum sanctum Linn.,also referred to as Tulsi, is a Southeast Asian Ayurvedic plant with a long history of
conventional use. The significance of this plant for use in food, medicine, and industry prompted researchers
to examine its chemistry and pharmacology.
Mangifera indica , commonly known as mango, is a pharmacologically, ethnomedically, and phytochemically
diverse plant. Various parts of M. indica tree have been used in traditional medicine for the treatment of different
ailments

Indigofera tinctoria is known as the indigo plant , is famous for its blue dye. It's been used for centuries
to produce indigo dye, which has been significant in textile production, especially in regions like
India.The indigo plant is also considered as a medicinal plant in the traditional sense to treat skin care

Chloroxylon swientenia is medicinal plant and finds immense

application as a phytopharmaceutical formulation for
therapeutic use of C.swietenia is a folklore medicinal plant that is
commonly used for antimicrobial, anti-fertility,analgesic
properties.
 angifera Indigofera Chloroxylon
Ocimum M
tinctoria swietenia
sanctum indica d )
(indigo) (s atinwoo
(Tulsi) (mango)
 Aim of the Project
Comparing the leaves of the plants Chloroxylon swietenia, Mangifera
indica, Ocimum sactum, and Indigofera tinctoria involves examining
factors such as their phytochemical properties, as they have potential
medicinal uses. Each plant may contain unique compounds with varying
pharmacological effects, such as anti- bacterial, antifungal, anti-
inflammatory, phenolic properties . The comparison aims to understand the
potential of each plant and their suitability for use in traditional medicine
or as sources of new pharmaceutical compounds.
Also comparing wild and cultivated (normal) plants involves
examining how different growing conditions and environments impact
the chemical composition, bioactivity, and potential medicinal
properties of plants, which help to identify active compounds within
plants, paving the way for the development of new drugs or natural
remedies.
 Acknowledgement

BioInnovale Osmania
Lifescience University
The results for test samples
The methodology used anti-inflammatory,
phenolics,and mic (minimum
in the entire project
inhibitory concentration)using
was guided by spectrophotometer was
bioinnovale life science provided by research college of
osmania
 Review of Literature
GC-MS Analysis and Antimicrobial Activity of Estimation of Total Phenols and
Phytochemical screening and Phytochemical screening and Flavonoids in Selected Indian
Phytochemical screening of Indigofera suffruticosa. antibacterial activity of quantitative estimation of total TraditionalPlants.DEEPAK KUMAR1
Indigofera tinctoria (Linn.) Evid based complement chloroxylon swietenia DC. Int J flavonoids of OCIMUM , ANUPAM JAMWALREECHA
Leaf Extract Characterizing alternat Med 2006 Jun; 3(2): MADAAN2
its Mediicinal use.
Pharma ResHealth Sci.2017 ; sanctum in different solvents
261–265.Published online and SURESH KUMAR
International Journal of 5(6),2002- extract. Praveen Garg and Department of Pharmaceutical
2006 Apr 5. doi: 07.DOI:10,21276.06.20,
Ayurvedic Medicine . Vol Rajesh Garg, ISSN (E):2277-7695 Sciences and Drug Research, Punjabi
10.1093/ecam/nel010PMCI CODEN(USA)-IJPRUR,e- University
11(2) , 289-299 , ISSN NO: , ISSN(P):2349-8242.TPI
D: PMC1475935 ISSN:2348-6465. RMahmood et Chitkara College of Pharmacy,
0976-5921.Mishra DN, 2019;8(2):16-21.
PMID: 16786057 Chitkara University, Rajpura-140 401,
Gomare KS, Sheelwant SV al . WWW.thepharmajournal.com Patiala, Punjab, India

Pharmacological evaluation of ANTIDERMATOPHYTIC ACTIVITY OF

Phytochemical and Pharmacological evaluation DIOSPYROS CHLOROXYLON
selected medicinal plants used
Pharmacological of medicinal plants with LEAVES: USING TWO IMPORTANT SOLVENT
in the management of oral and EXTRACTS
evaluation of ethanolic antidiabetic activities in skin infections in Ebem-Ohafia 1 2* P. SHIVAKUMAR SINGH AND G.M.
VIDYASAGAR
extract of Lepisanthes Ethiopia:A District , Abia state,Nigeria. 1Department of Botany, Plamuru University,
rubiginosa L.leave. hasan rewiew.Metabolism Open 13 Hindawi, The scientific world Mahabubnagar-509001, Telangana, India.
2Medicinal Plants and Microbiology Research
et al.BMC Complemetary (2022) 100174. Journalvolume 2018 , Article ID
Laboratory Department of Post- Graduate
4757458, 16
and Alternative Medicine www.sciencedirect.com/jour Studies and
pages.http://doi.org/10.1155/201 Research in Botany Gulbarga University,
(2017) 17:49. DOI 10.1186/s nal/metabolism-open 8/4757458 Gulbarga – 585 106, Karnataka, India.
*Corresponding author:
12906-017-2010-Y gm.vidyasagar@rediffmail.com
 Materials and Methods
Sample Preparation

Collection and Filtration of Then, it was filtered

pondering of leaves samples with Whatman No. 1

filter paper and the
The leaves of The solution was left solvent part was
for 24 hrs collected
satinwood,
at room temperature
Tulsi,indigo,mango
were collected , dried
and powdered.
Preparation of 5grm of plant extract was
solvents added in 100ml of
DISTILLED WATER
The plant extract
was added to 4 5grm of plant extract was
added in 100ml of ACETONE
different solvents
in the ratio of 1:20
5grm of plant extract was
added in 100ml of ETHANOL

5grm of plant extract was

added in 100ml of
HYDROALCOHOL( 60%ETHANOL
 Screening of plant secondary metabolites

We have done 12 tests to find out secondary metabolites in the plant

REAGENT PREPARATION FOR PHYTOCHEMICAL SCREENING

1.Wagner's reagent: Dissolve 2 g of lodine and 6 g of

1.Wagner's reagent: Test for Alkaloids
Potassium iodide in 100 ml of water. 2 Molish's test: Test for Carbohydrates
2 Molish's test: Disslove 15g of a-naphthol in 100 ml of 3. Keller kelliani's test: Test for Cardiac
alcohol or chloroform.
Glycosides
3. Keller kelliani's test: Glacial acetic acid, 5% aqueous ferric
chloride, and conc. Sulphuric acid. 4. Alkaline reagent: Test for Flavonoids
4. Alkaline reagent: 20% Sodium hydroxide solution. (NaOH) 5. Aqueous Ferric chloride: Test for Phenols
5. Aqueous Ferric chloride: 5% aqueous Ferric chloride
6. Ninhydrin Tests : Amino acids and Protein
solution.
6. Ninhydrin Tests 1% solution of Ninhydrin in acetone. 7. Foam Test : Test for saponins
7. Foam Test (Saponin) and Turbidity Test (Resin): Distilled 8. Alcoholic Ferric chloride: Test for tannins
water.
9. Salkowski's test: Test for terpenoids
8. Alcoholic Ferric chloride: 10% Ferric chloride in absolute
alcohol solution. 10. Coumarins: presence of coumarins
9. Salkowski's test: Chloroform and concentrated Sulphuric
acid
10. Coumarins: 10% Sodium Hydroxide (NaOH).
 QUANTITATIVE ESTIMATION OF PHENOLIC by F/C method
Preparation of standard
Requirement: Plant extract, Distilled water, Gallic acid (Standard), F/C reagent, Sodium Carbonate
(Na2CO3). 100 mg of the extract of the sample was weighed and dissolved in 100 ml of triple distilled
water. 1 ml of this solution was transferred to a test tube, then 0.5 ml 2N of the Folin-Ciocalteu
reagent and 1.5 ml 20% of Sodium carbonate solution was addor 3) Es volume was made upto 8 ml
with Triple Distill Water followed by vigorous shaking. This was allowed the total 2 hours. The
absorbance was measured at 765nm in a balibrated UV-Visible Spectrophotometer. The total
phenolic content was estimated using a standard calibration curve of gallic acid. (10 µg/ml,
200mg/ml, 80 µg/mlg

Preparation of test samples

0.1 ml of plant extract , 0.1 ml gallic acid, 0.3ml of AlCl3, 4ml of double distilled water, 2ml of NaOH was
added to the test tube and incubated at room temperature for 5 mins. Then 0.3ml of NaNO2 was added
to the test tube. Make the sample to final volume of 10ml . The sample was tested at UV light Visible
Spectrophotometer at 765nm. A standard curve of absorbance against gallic acid concentration must
be prepared which is used for estimation of total phenols content in test samples.
Screening of plant secondary metabolites

QUANTITATIVE
ESTIMATION OF
PHENOLIC by F/C
method
 Anti microbial activity of bacteria and fungus
●For preparation of bacteria , the nutrient broth was prepared by adding 4g of nutrient broth powder in
250ml of distilled water and was heated in hot plate. Then the microbial culture ( e.coil)was transferred to
the using inoculation loop and was incubated for 24hr for formation of culture.

●For preparation of fungi culture nidulus was used, PDA broth was prepared and the nidulus was subculture in
broth and was incubated in orbital shaker for 24hr
●For media preparation for bacterial culture, Nutrient broth of 1 lit was prepared then 9 grms of agar agar was
the broth and heated on hot plate for 15 mins. For media preparation of fungus, 1 lit PDA broth was prepared a
grms of agar agar was added and heated in the hot plate for 15 Mins and the media was autoclaved at 121°C,15
15mins
● Petriplates was double sterilized in autoclave and hot air oven . The laminar air flow was sterilized using stan
● The respective media was poured onto the plate and allowed to solidified, 1 ml of liquid microbial culture
was poured into the respective media and was spread using a spreader. 5 wells were made in each petriplate,
one in the center( negative)and other 4 in the top, bottom , left and right which marked as P1, P2,P3,P4.

● Using micropipette, in negative distilled water was filled in the wells, In P1 Hydro alcohol was filled in wells of
Chloroxylon and mango and in P1 Ethyl alcohol was filled in the wells of Tulasi and indigo. In P2 Ethyl alcohol was
filled in the wells of Chloroxylon and Mango whereas in tulsi and Indigo P2 is Hydroalcohols solvent. In P3 & P4
distilled water solvent and acetone was filled in the wells of respective plant petriplates
Anti microbial activity of bacteria and fungus
In-vitro Pharmacological study. Anti-inflammatory assay
(Inhibition of Protein denaturation)
Requirement: Plant extract, Egg albumin (1%), Phosphate buffer-saline (PBS, pH-6.4)

The reaction mixture (5 mL) consists plant of extract 0.1ml and the mix bovine egg albumin 0.2ml , 4.78 mL of
phosphate buffer and the mixture was mixed, and was incubated in a incubator at 37C for 15mins and then the
reaction mixture was heated at 70 "C for 5 min. After cooling, the turbidity. The measured at 660 nm in
spectrometer and phostpahe buffer was used as cobtrolwas used as the control. The of protein denaturation
was calculated by using the following
Absorbance control - Absorbance test
% of denaturation = × 100
Absorbance control
 Result Analysis

✅ ✅ ✅ ✅ ✅ ✅ ✅ ✅ ✅
✅ ✅ ✅ ✅ ✅
✅ ✅ ✅ ✅
✅ ✅
✅ ✅ ✅ ✅ ✅ ✅ ✅ ✅ ✅
✅ ✅ ✅ ✅ ✅ ✅
✅ ✅ ✅
✅ ✅ ✅ ✅ ✅ ✅ ✅
✅ ✅ ✅ ✅
✅ ✅ ✅ ✅ ✅ ✅ ✅

Screening of plant secondary metabolites

 Screening of
plant
secondary
1 1 1 1 1 1 metabolites

1.Satinwood
1 1 1 1 1 1
2.Mango
3.Indigo
4.Tulsi
2 2 2 2 2

2
2 3 3 3 4 4
QUANTITATIVE ESTIMATION OF PHENOLIC by F/C method

Concentration Absorbance Cat (415nm)

10 0.02
20 0.04
40 0.06
60 0.10
80 0.12
100 0.16

Result of standards
 Antimicrobial activity:

Mangifera indica Octimum Sanctum

Indigofera tinctoria Chloroxylon
P1: Ethyl. alc Hydro.alc
P2: Hydro.alc Ethyl.Alc E.coli showed inhibition in:
Mangifera indica- Distilled water (P3)
P3: Distilled water Distilled water
Indigofera tinctoria-Hydro.alc (P2)
P4: Acetone Acetone
Octimum Sanctum- Ethyl alc (P1)
Tulsi / chloroxylon/ Chloroxylon- Ethyl.alc (P2)
indigo Mangifera indica
 Anti bacterial
Satinwood activity

Mango

Tulsi
Indigo
Antifungal activity

Chloroxylon

Mango
 ANTIMICROBIAL ACTIVITY IN BACTERIA(E.COLI )

Hydro alcohol Ethylalcohol Distilled water Acetone

Mango Nil Nil 10

Nil
Maximum inhibition is
mm

11
seen in chloroxylon with
Satinwood Nil Nil Nil
ethyl alcohol in E.Coli
mm

Tulsi Nil
bacteria
10mm
Nil Nil

Nil
Indigo 8mm Nil Nil

ANTIMICROBIAL ACTIVITY IN FUNGAL( NIDULUS )

Hydro alcohol Ethylalcohol Distilled water Acetone

Maximum inhibition is seen
Mango
in chloroxylon with ethyl
Nil 9 mm Nil Nil

13
Satinwood Nil mm Nil Nil
alcohol when done using
fungus nidulus
 FUTURE WORK
In-vitro Pharmacological study- MINIMUM INHIBITORY CONCENTRATION (MIC)

1.Nutrient broth was prepared and autoclaved at 15 psi pressure in 121°C for 15 minutes.

2.8 test tubes were taken( auto claved). To each 8 test tubes, 5 ml of autoclaved Nutrient broth was poured.

3. All the test tubes were then inoculated by 100µl. of microbial suspension culture.

4. One test tube is the kept as positive control and one test tube is kept as negative control.

5. Amongst six test-tubes, the plant sample was added in the decreasing concentration as 1000µg/ml,
800µg/ml, 600µg/ml, 400µg/ml, 200µg/ml, 100µg/ml respectively to the six inoculated test tubes.

6. These test tubes were then were kept for incubation in the incubator at 37°C for 24-36 hours.

7. After incubation the suspension broth was spectrophotometrically analysed at 600 nm with water kept as
blank. The graph was plotted using absorbance against concentration of the plant extract.
QUANTATIVE ANALYSIS OF PHENOLIC
Samples are given to the lab

ANTI-INFLAMATORY:
Samples are given to the lab
THESIS:
We have started writing and will be submitted on time.
 CONCLUSION
Overall,each plant extract has shown varied outcome on conducting phytochemical
analysis, anti microbial test on both bacteria and fungus.
In photochemical analysis different tests like alkaloid,terpenoid etc are done to which
each plant extract reacted differently and few shown positive response and others have
shown no effectt.
Quantitative estimation of phenolic by f/c method was done on all plant extracts to which
results are yet to come .
Anti microbial test with E.Coli bacteria was performed to see the level of inhibition to
which maximum inhibition is seen in chloroxylon with ethyl alcohol,
Aslo anti microbial using fungus was also conducted on mango and chloroxylon to which
again chloroxylon showed maxmimum inhibition in ethyl alcohol.
Anti inflammatory are also conducted to which results are yet to come as we gave it to a
lab for the final result using spectrophotometer
And we are left with anti diabetic test that will also be conducted on all four plant extracts.
Thank's For
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