Our Study was aimed at evaluating the anti-inflammatory of activity Clerodendrum Inerme , Methanolic Root Extract. We evaluated under the HRBC Membrane Stabilization and Protein Denaturation Method.

1. PHYTOCHEMICAL SCREENING AND INVITRO ANTI-INFLAMMATORY ACTIVITY OF
METHANOLIC ROOT EXTRACT OF Clerodendrum Inerme.,
Presented by
Naveen Kumar S (RA1621003010088)
Dyari Sherko Mustafa (RA1621003010076)
Praveen Kumar S (RA1621003010081)
Bewar Tahsin Karim (RA1621003010081)
Nwafor Obasi Ivuoma Roseline (RA1621003010081)
Surendhar V (RA1621003010081)
Under the guidance of
Dr. M. Thirumal, M. Pharm., Ph.d.,
1

2. Introduction
 The word inflammation is derived from the Latin word “inflammo” which means
I set alright, “ignite”. Inflammation is a complex biological response of vascular
tissues to harmful stimuli like infection, irritation, pathogen damaged cell,
irritants or foreign substances.
 Inflammation is basically of two types
2

3. Introduction
 Acute Inflammation : It is achieved by increased movement of plasma and
leukocyte present in the blood towards the injured tissues which is due to the
initial response made by the body towards the harmful stimuli and chronic
inflammation or the prolonged inflammation is characterized by progressive shift
in types of cells found at the inflammation site which undergoes simultaneous
destruction following with the healing of tissues due to inflammation process.
 Chronic Inflammation : Includes asthma, chronic peptic ulcer, rheumatoid
arthiritis, tuberculosis, ulcerative colitis and chronic hepatitis, chronic sinusitis. In
the mechanism of acute inflammation it starts with tissue injury within a few
seconds or in a few minutes.
3

4. PHARMACOGNOSY & PHYTOCHEMICAL :
 Arshad Farooqul, MD et al.(2010)- have extracted silver nano particles using extracts of leaf Clerodendrum inerme.
 Gurudeepan, X et al. (2010)- have worked on antioxidant and radical scavenging effect of Clerodendrum inerme and
concluded the anti-oxidant property of Clerodendrum inerme was due to phenolic compounds.
 Jasvinder Kaurchahlal, et al. (2011)- have studied on efficacy of Clerodendrum inerme against human pathogenic strains. The
methanolic extract of the leaves shows higher inhibition against S.aureus and A.riger with decreased MIC values 0.78 mg/ml.
 Kothari Avani et al.10 (2006)- have showed ex situ conservation method for Clerodendrum inerme.
 Margaret beula Banerjee al. (2012)- have carried out the studied on antiviral, anti-oxidant and toxicological evaluation of
mangrove associate from south east coast of india. Evaluations are done for Salicornia brachiate, Clerodendrum inerme,
Rhizopharalamarckii.
 Prasad M.P et al. (2012)- have studied phytochemical analysis, anitoxidant potential, anti-bacterial activity & molecular
characterization of Clerodendrum inerme. Species reported the chemical constituents in 6 species of (Clerodendrum inerme,
pariculatum, Phiippinum Phlomoids, serrattum, vilosum). The study revealed the anti-oxidant activity shown to be maximum
in Clerodendrum inerme than other species.
 Triptech Kanchanapoom et al. (2001)- have elucidated that megastigmane and irridiod glycosides from Clerodendrum inerme.
Review Of Literature
4

5. PHARMACOLOGICAL :
 Graimaupamyu, et al.(2011)- have carried out the acute toxicity and diuretic activity in male albino wistar rats. The extract of leaves of
Clerodendrum inerm showed good diuretic activity in wistar rats after 24 hrs by increase in potassium and sodium ion excretion
 Guessan, KN et al. (2009)- have worked on Hypotensive effect of aqueous extract of Clerodendrum inerms leaves on the arterial
pressure of rabbit and reported that the dried leaves of plant Clerodendrum inerme found to have hypotensive effect at higher doses
where as lower concentration has no effect on blood pressure.
 Rajsekran Anitha & Ponnusamy Kannan, et al. (2005)- have studied the anti-fungal activity of Clerodendrum inerme and phlomoids.
The thyl acetate extract of leaves was reported for anti-fungal activity.
 Suresh, M et al.- have studied anti-fungal activity of selected indian medicinal plant salt and found that plant salt of indian medicinal
plants. i.e, Acalypha indica, Centellaasiatica, Eervalanata, Clerodendrum inerme, Pergularia daemia, Solamammelo genaadd promising
anti-fungal activity against candida Albicans, Cryptococus Meoformans and Aspergillus Flavus the anti-fungal assay was performed
using agar disc diffusion method and it was selected medicinal plant revealed to posses potential anti-fungal activity.
 Vijayamirtharaj,R, et al. - have studied anti-inflammatory and analgesic properties of methanolic extract of aerial part of Clerodendrum
inerme in experimental animal model, reported that the methanolic extract of aerial part, at dose of 200 mg/kg body weight produced
significant analgesic and anti-inflammatory.
Review Of Literature
5

6. Aim
The present study was aimed at evaluating the anti-inflammatory activity of
Clerodendrum inerme methanolic root extract.
Objectives
To do the phytochemical screening and Pharmacological evaluation by
HRBC membrane stabilization and protein denaturation method.
6
Aim And Objective

7. Materials And Methods
Plant Collection and Authentication :
 Fresh roots of Clerodendrum inerm (L.) Gaertn. Verbanaceae (Laminaceae) were collected from the
region of Avadi, Chennai, Tamilnadu. The plant was identified, and authenticated by comparing with
an authentic specimen by botanist Dr. P. Jayaraman, Plant Anatomical Research center (PARC),
Tambaram, Chennai
 The extract was then concentrated by distilling of the solvent and evaporated to dryness and the
percentage yield was calculated.
Preparation of Plant Extract :
The powedered root (500g ) of Clerodendrum inerme was extracted with methanol using soxhlet apparatus for
72 hours on a water bath. The extract was evaporated to dryness on water bath to obtain semi-solid mass. The
dried extract of root of Clerodendrum Inerme was collected and stored in the refrigerator until used for further
studies.
7

8. Preliminary Phytochemical Analysis
TEST FOR CARBOHYDRATES :
MOLISH’S TEST (GENERAL TEST) : To 2-3ml aqueous extract, add few drops of alpha-napthyl solution in
alcohol, shake and con. Sulphuric Acid from sides of the test tube. Violet ring is formed at the junction of two
liquids.
TEST FOR REDUCING SUGARS:
FEHLING’S TEST : Mix 1ml Fehling’s A and 1ml Fehling’s B solutions, boil for one minute. Add equal
volume of test solution. Heat in boiling water bath for 5-10minutes first a yellow, then brick precipitate is
observed.
BENEDICT’S TEST : Mix equal volume of Benedict’s reagent and test solution in test tube. Heat it in boiling
water for 5 miutes, Solution appears green, yellow, or red depending on amount of reducing sugar present in test
solution.
8

9. Preliminary Phytochemical Analysis
TEST FOR MONOSACCHARIDES :
BARFOED’S TEST : Mix equal volume of Barfoed’s reagent and test solution. Heat for 1-2 minutes in boiing
water bath and cool. Red Precipitate is observed.
TEST FOR PENTOSE SUGARS :
 Bail’s Orcinol Test : To boiling Bail’s reagent add few drops of test solution. Green or purple coloration
appears.
 Aniline Acetate Test : Boil test solutions in test tube. Hold filter paper soaked in aniline acetate in the
vapour. Filter paper turns pink.
TEST FOR MUCILAGE :
 Powdered drug material shows red color with ruthenium red.
 Powdered drug swells in water or aqueous KOH. 9

10. Preliminary Phytochemical Analysis
TEST FOR GLYCOSIDES :
Determine free sugar content of the extract. Hydrolyse the extract with mineral acid. Again determine the total
sugar content of the hydrolysed extract. Increase in sugar content indicates presence of glycosides in the extract.
Glycoside = Aglycon + Glycon
TEST FOR SAPONIN GLYCOSIDES :
 Foam Test : Shake the drug extract or dry powder vigorously with water. Persistent foam is observed.
 Haemolytic Test : Add drug extract or dry powder to one drop of blood placed on glass slide. Haemolytic
Zone appears.
10

11. Preliminary phytochemical screening
 The variety of plant and plant extracts contains different phytochemicals with biological
activity that can be of valuable therapeutic Index. The much of the protective effect of fruits
and vegetables has been attributed by phytochemicals which are the Non-nutrient plant
compounds.
 Different phytochemicals have been found to possess a wide range of activities,
which may help in protection against chronic diseases. The results obtained in the present
investigation are reported in the Table.
Result
11

12. Preliminary phytochemical screening
Result
12
The results obtained in the present investigation are reported in the Table
Phyto Constituents Methanolic Extract
Alkaloids +
Glycosides +
Proteins +
Cardiac Glycosides -
Flavonoids +
Glycosides +
Phenols +
Saponins +
Steroids -
Tannins +
Amino Acids +
+ = Presence of
Phytoconstituents
- = Absence of
Phytoconstituents

13. Preliminary phytochemical screening
Result
13
HRBC Membrane stabilization Method :
It was observed that methanolic extract of root of
Clerodendrum inerme had lower activity than that
of standard diclofenac sodium. At a concentration
of 1000mcg/ml, the anti-inflammatory activity of
standard Diclofenac sodium reached 94.66%
while at the same concentration, methanolic
extract of Clerodendrum inerme root was 85.34%
.Though the HRBC membrane stabilization
abilites of the extract were less than those of
Diclofenac sodium as standard. When observe the
IC 50value of methanolic extract of
Clerodendrum inerme is 142.86 μg/ml and in
same way for standard is 91.24 μg/ml.

14. Preliminary phytochemical screening
Result
14
Inhibition of Protein Denaturation Method:
It was observed that methanolic extracts of root
Clerodendrum inermehad lower activity than
that of standard Diclofenac sodium . At a
concentration of 1000mcg/ml, the arthritic
activity of standard Diclofenac sodium reached
94.82% while at the same concentration
ethanoilc extract of Clerodendrum inerme root
was 80.82% . Though the protein denaturation
abilites of the extracts were less than
Diclofenac sodium. When observing IC 50
value of the ethahoic extract of Clerodendrum
inerme is 142.65 μg/ml and in same for
standard is 108.86 μg/ml.

15. Discussion And Conclusion
 In the present study, we have found that methanolic extract of Clerodendrum inerme
root is having significant anti-inflammatory and anti- arthritic activity.
 It shows the significant effect in concentration dependent manner. The activity may be due to the
presence of phytoconstituents such as phenol, steroids, di and tri terpenes, volatile oils, alkaloids,
glycosides, carbohydrates, flavonoids, hydrocarbons, proteins, phytosterols, quinines, saponins.
 This work provides an insight to understanding some molecular basis of therapeutic
properties of Clerodendrum inerme. Our future aim is to isolate and characterize the chemical
constituents responsible for the anti-inflammatory and anti arthritic activity.
 We strongly believe that the outcome of the study will trigger exciting research on
addressing the anti-inflammatory and anti-arthritic drug in a cost effective manner.
15

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Thank You