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PHYTOCHEMICAL AND EVALUATION OF ANTI ULCER EFFECT OF CORDIA
DICHOTOMA G (ForsT
...

A Dissertation Submitted to

JAWAHARLAL NEHRU TECHNOLOGICAL UNIVERSITY,
HYDERABAD
...
T
...
15S61S0103
Under the Guidance of
Mr
...
Shamim Qureshi M
...


Associate Professor
DEPARTMENT OF PHARMACOGNOSY
ANWARUL ULOOM COLLEGE OF PHARMACY
NEW MALLEPALLY, HYDERABAD- 500 001
...
(ForsT
...

Peptic/ stomach ulcer is the
condition in which imbalance of
aggressive factor and defensive
factors
...

Defensive factor: mucous and
Bicarbonate secretion
...


PEPTIC ULCER
CAUSES
� Decreases mucous secretion
and bicarbonate secretion
� Increases protein diet
� Skipping of diet
...

� Alcohol
...

�

eat

�

DIAGNOSIS
...

Endoscopic biopsy:
...
and it also grow in outer tract of
sub Himalaya ranges ,which is having elevation of about 1500
m in ascending and it is also native in India and Nepal
...
The plants with various parts
like leaves, roots, seed, bark, and fruits which shows the
various activity like immunodulator, antidaibetic
...
– Cordia
Species: Cordia dichotomaG
...
– fragrant manjack
o

VERNACULAR NAMES
o

o
o

Hindi
–
Gunda
or
Tentidela
...

English: Fragrant manjack
...


Odour

Taste

Shape

• None or pleasant

Fruits are sweetish and
mucilaginous

•Leaves are
ovate
...

▪ Africa and
▪ Asia
...

Antifungal
� larvacidal
...

� Cytotoxic
...

� Anti-inflammatory
� Anthelmintic
� Wound healing
�

AYURVEDIC USES
...

⮚ The bark and fruits of
the tree are used in
treatment of variety of
diseases
...


AIMS AND OBJECTIVES
...


PLAN OF WORK
Collection, Identification and Authentication of plant
...
e
...

Extraction of plant material
...


Preliminary Phytochemical screening
...
(Leaves & Root Bark)

Anti-Ulcer Activity
Methanolic & Aqueous
extracts
Acute &chronic model
Aspirin induced
gastric ulcer in Rats

Acute & chronic model
Ethanol induced gastric
ulcer in Rats

METHODOLOGY

800 g
...

to
hot
Mahasamund,
continuous
Chhattisgarh
...
Soxhlet
obtain
apparatus
Ahmedullah,
ed
with
Scientist
“E”
from - in
solvents
Botanical
thethe
survey
CPCS of
increasing
India,
Ministry
EA/IA
of
oforder
EC
polarity
Environment
for
&using
Forests,
anima
petroleum
Deccan
l
ether,
regional
studie
chlorofor
Centre,
m,s
Hyderabad
...
S
...

for
ration
extract
Cordia
No
...

controlled
BSI/DRC/2015

Drying of
leaves

Collection And
Authentication
Of Plant
Materials

CPCSEA /
IAEC
Approval for
Animal
Studies

COLLECTION
AND AUTHENTICATION

Soxhlet Extraction
�

Aim: To perform the Soxhlet extraction with different solvent according to polarity

like boiling point
...
First the leaves are weighted and inserted in a Soxhlet
apparatus and in Soxhlet first the cotton is inserted and then the
powder of leaves are poured in a Soxhlet and then the Soxhlet is fixed
to the RBF and heating mantle and then pet –ether is poured in a
Soxhlet and then the condenser is placed which is attached to the pipes
and motor and the temperature of heating mantle is maintain at35°c
and then steam distillation is occurred and siphon take place with the
help of siphon tubes and then the final extract was formed in the RBF

Percentage Yield
The percentage yields of the extractives were calculated using the
formula:
Weight of the extractives
% Yield =------------------------------------ x 100
Weight of the crude drug

PHYTOCHEMICAL
SCREENING
PHYTOCHEMICAL SCREENING TEST

S
...


NAME OF THE TEST

1
...
H2SO4 From The Sides
Junction
Of The Test Tube
...


Fehling’s Test

First Yellow, Then
Brick Red Ppt
...


Presence Of Reducing
Sugars
...


Benedict’s Test

Take Equal Volumes Of
Fehling’s A And B Reagents,
Add 2-3 Ml Of Test Solution
And Heat On A Water Bath For
5 Min
...


A
...


Test For Cardiac Glycosides:
Baljet’s Test:

Thin section of the drug is dipped in Sodium Yellow to orange colour is
picrate solution
obtained
...


a
...


Legal’s Test

To aqueous or alcoholic extract add 1ml
...

Pyridine and 1ml
...


Presence of Cardiac glycosides
...


Keller Killiani Test

To 2ml
...
H2SO4
...


Presence of Cardiac glycosides
...
of the extract add dil
...
Cool the filtrate and shake with equal
volume of CHCl3

Ammonical layer does not turn to Absence of Anthraquinone glycosides
...


3
...


b
...

a
...

a
...
Hcl
...
5 gm of magnesium
observed
...


b
...
of NaOH sol
...
Hcl
...
later changes to colourless
...


Zinc Hcl Test

Treat the extract with Zinc dust and conc
...


To 5 ml extract add 5ml 5% FeCL3 and 5ml
Ammonical layer shows pinkish- Presence of anthraquinone glycosides
dilute Hcl
...


Presence of saponin glycosides

Presence of Flavonoids
...


6
...


Test For Alkaloids:
Dragendroff’s Test

b
...
, add Dragendroff’s Orange brown ppt
...


Presence of Alkaloids

Mayer’s Test

To the test sol
...
is formed
...


c
...
, add Hager’s reagent

Yellow ppt
...


Presence of Alkaloids
...


Wagner’s Test

To the test sol
...
is obtained

Presence of Alkaloids
...


Murexide Test For Purine Alkaloids

Presence of Alkaloids
...

a
...
test solution add 3-4 drops Purple colour is observed
...
HNO3, evaporate to dryness,
cool and add two drops of NH4OH sol
...


Presence of Tannins

b
...

Vanillin Hcl
...


Presence of Tannins

c
...


Discoloration of the extract is Presence of Tannins
observed
...


Acetic Acid Solution Test

Treat the extract with Acetic acid sol
...


Presence of Tannins
...


Lead Acetate Solution Test

Treat the extract with Lead acetate sol
...
is formed
...


f
...


White ppt
...


Presence of Tannins
...


Potassium Dichromate Test

Treat the extract
dichromate sol
...


Dilute Iodine Solution Test

Treat the extract with dilute iodine Translucent red colour is seen
solution
...


i
...

solution

Presence of Tannins
...


Dilute
Nh4oh
And
Ferricyanide Solution Test

with

Potassium Treat the extract with
Dilute
NH4OH
and
ferricyanide sol
...
appears

Potassium

Red color appears
...


ASPIRIN INDUCED
GASTRIC ULCER
⮚

⮚

⮚

⮚

�

�

Group-1 (control) treated with Normal
(Distilled water)
Group-2 Standard received Ranitidine
(Ranitidine 2mg/kg, b
...
o) for once
daily for 10 days
Gropu-3 (Test-1) received CDLME
(250mg/kg b
...
o) once daily for 10
days and Aspirin (250mg/kg, b
...
o)
suspended in 1% CMC on 10th day
...
w, p
...
w p
...

Gropu5-(Test-3) received CDLAE
(250mg/kg b
...
o) once daily for 10
days and Aspirin (500mg/kg, b
...
o)
suspended in 1% CMC on 10th day
...
w, p
...
w p
...


Male and female rats is taken which is having
weight approximately 160g were used for the
purpose of study
...
But free access to water, the
aspirin were suspended in (500mg/kg, b
...
o) which was suspended in 1% CMC of
carboxyl methyl cellulose at a concentration
of 50mg \ml
...
After a time interval of one,
two, three four up to tenth days
...
Then the surface
of the mucosa was wiped with a soft tissue
...


ETHANOL INDUCED
GASTRIC ULCER
�
�

�

�

�

�

Group-1 control (Distilled water)
Group-2
Standard
(Ranitidine
2mg/kg, b
...
o in 1 ml of 1% CMC
Group-3 (Test-1) received CDLME
(250mg/kg b
...
o) once daily for 10
days and Ethanol (1ml/kg b
...
o)
on 10th day
...
w, p
...
w, p
...

Group-5 (Test-1) received CDLAE
(250mg/kg b
...
o) once daily for 10
days and
Ethanol (1ml/kg
b
...
o) on 10th day
...
w, p
...
w, p
...


First the rats are kept in a fasting stage for
24 hrs after competition of 24 hrs the
rats are treated with 1 ml of ethanol
through orally
...
and then the drugs are
administrated orally in 30 min before the
administration of ethanol 1ml
...

Then the gastric contents were subjected
to centrifuge to centrifugation at 1000
rpm for 10 mins
...


Collection and Measurement of Gastric juice
...
Then the gastric
content were collected and centrifuged at 1000 rpm for 10mins
...

� Determination of pH of Gastric juice
...
With the help of digit meter
...

� Estimation of Free and total Acidities
...
In that2 to 3 drops of
Tofers reagent was added to the conical flask and then titrate with 0
...
The volume of alkali is noted
...
then 2 or 3
drops of phenolphthalein solution was added and the titration was continued until a definite
reading appears
...
This volume correspond
total acidity the total acidity was calculated by using the formula
...
ofNaOH×N×100mEq/L
Acidity =
-----------------------------------------0
...

Mean ulcer score for each animals is expressed as Ulcer Index
...

The number of ulcer present in the stomach were noted and severity of the scoring
was done with the help of microscopically and score method was done with the help
of as per
...
0
...

1
...

2= Ulcer>3mm but<5mm3=Ulcers>5mm
Ulcer index (UI) = UN+US+UP×10
Where, UI = ulcer index
UN = average number of ulcers per animals,
US = average of severity score
UP = percentage of animals with ulcers
...
19 Flow chart Representation of Anti Ulcer activity (Including Acute and Chronic model)

�

�

�

PLANT MATERIAL AND EXTRACTION
The percentage yield of the successive leaf extractives of Cordia dichotoma are present in the
table
...
) Leaves Methanaolic Extract by
Soxhlet Extraction and Aqueous Extraction by Decoction method was calculated and the results
obtained are as follows:
Table 6 Extractives
Percentage Extraction result
value of Cordia
G (ForsT)
leaves analysis
Nature
Yielddichotoma
(%w/w)
Phytochemical

Petroleum

Greenish brown solid

0
...
5% w/w

Alkaloids, phenols ,terpenoids

Methanol

Brown solid

2
...
8 %w/w

Flavonoids, phenolenoids, tannins

�

Phytochemical Screening:
The results of the Preliminary Phytochemical Screening of the Cordia dichotoma Methanaolic and Aqueous Extracts are
as follows:
Results of Phytochemical Screening of Cordia dichotoma G
...


Test For Alkaloids
Tannins

Test For Flavoids

Test For

Test For Amino Acids Test For Steroids Triterpenoids Test For Carbohydrates Test For
Alkaloids

RESULT OF GASTROPROTECTIVE EFFECT:
TABLE 8- Result of gastro protective effect of CDLME and CDLAE on Volume of Gastric Juice, pH, Free Acidity, Total
acidity, Ulcer index and % Ulcer protection in Experimental model No
...

S
...

b
...
p
...
w, p
...
of 1%CMC
+CDLME(250mg/kg b
...
o in 1
ml of 1% CMC)
Test-2 Aspirin 500mg/kg b
...
o
in 1ml
...
w, p
...
w, p
...
of 1%CMC
+CDLAE(250mg/kg b
...
o in 1
ml of 1% CMC)
Test-2 Aspirin 500mg/kg b
...
o
in 1ml
...
w, p
...
52±0
...
6±0
...
25±0
...
3±0
...
3±0
...
2±0
...
2±0
...
19±0
...
95±0
...
95%

5
...
09*

5
...
12*

41
...
26

65
...
06

4
...
43*

71%

4
...
12**

6
...
08**

43
...
18

55
...
33

2
...
08

75
...
7±0
...
9±0
...
8±0
...
1±0
...
10±0
...
28%

47
...
16**

6
...
10**

41
...
97

48
...
88

3
...
42**

68
...
2 Ethanol
Induced Gastric ulcers in Albino wistar Rats
...
6±0
...
85±0
...
0±0
...
13±1
...
3±0
...
7±0
...
08±0
...
2±0
...
2±0
...
5%

Test-1 Ethanol (1ml/kg b
...
o in 1ml
...
w, p
...
3±0
...
49±0
...
8±0
...
94±0
...
5±0
...
5%

Test-2 Ethanol (1ml/kg b
...
o in 1ml
...
w, p
...
79±0
...
3±0
...
66±0
...
83±0
...
30±0
...
70%

Test-1 Ethanol (1ml/kg b
...
o in 1ml
...
w, p
...
8±0
...
49±0
...
0±0
...
66

7
...
11*

58
...
w, p
...
of
1%CMC) +CDLAE(500mg/kg b
...
o in 1
ml of 1% CMC)

2
...
25**

3
...
19**

22
...
93

39
...
31

6
...
11**

62
...
NO

TREATMENT

1

Control
(Distilled water)
Standard (Ranitidine2mg/kg
...
w
...
o in 1
ml of 1%CMC)

2

3

4

5

6

Morphological Appearance of Gastric Ulcers in Aspirin Induced Gastric
Ulcers

• Control

• Standard
• Test -1 (CDLME 250mg/kg b
...
w)
• Test -3 (CDLAE 250mg/kg b
...
w)

�
�

�

Histopathological Studies
Stomachs were preserved in 10% formaldehyde samples were given to shadan institute of
Medical sciences and research center for sectioning, staining and assessing Histopathological
studies
...
Section of 5-6 microns
in thickness were cut and stained with staining reagents
...

Plate Histopathological Studies of the Antiulcer activity using Aspirin Induced Model of
Cordia dichotoma CDLME (250mg/kg, b
...
Phytochemical constituents like tannins,
Flavonoids, alkaloids, glycosides are reported for their
anti-ulcer activity
...
Cordia Dichotoma Ger (ForsT)
Leaf Methanaolic and Aqueous extracts suggests to
be beneficial in antiulcer disorders
...
Herbal medicine
...

Dubey NK, Kumar Rajesh, Tripati Pramila
...
Current Science 86:2004; 37-41
...
Pathological Basis of disease, 6th Ed
...
B
...

Bose Manoj, Motgere, Vijay M, Dakhale, Ganesh N, Tarunka Avinash
...
Pol J Pharmacol 55:2003; 91-95
...
Role of
oxygen radicals in ischemia-induced lesions in the cat stomach
...




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