Clerodendrum volubile P Beauv (Verbenaceae) is a climbing shrub used traditionally in the treatment of swelling, gout and venereal diseases.  In this study, the microscopic, macroscopic, phytochemical analysis and anti-inflammatory activities were investigated using standard procedures. The results revealed the presence of alkaloids, flavonoids, saponins, anthraquinone and cardiac glycoside while the methanol extract, petroleum ether and ethyl acetate fractions exhibited significant (p&lt;0.05) anti-inflammatory activities when compared to the effect of the reference drug Diclofenac sodium

ADEKOYA, YEWANDE

JAIYESIMI, ADEDIWURA

English

Advanced Research Journals

  
 
International Journal of Phytomedicine 4 (2012) 414-418 
http://www.arjournals.org/index.php/ijpm/index 
 
Original Research Article 
             
Pharmacognostic Studies and Antiinflammatory Activities of Clerodendrum 
Volubile P Beauv Leaf 
Fred-Jaiyesimi Adediwura1* and Adekoya Yewande1 
 
*Corresponding author: 
 
Fred-Jaiyesimi Adediwura 
 
1Department of Pharmacognosy, 
Faculty of Pharmacy 
Olabisi Onabanjo University, Sagamu 
Campus Ogun State, Nigeria. 
 
 
 
 
 
 
 
A b s t r a c t  
Clerodendrum volubile P Beauv (Verbenaceae) is a climbing shrub used traditionally in the treatment 
of swelling, gout and venereal diseases.  In this study, the microscopic, macroscopic, phytochemical 
analysis and anti-inflammatory activities were investigated using standard procedures.   
The results revealed the presence of alkaloids, flavonoids, saponins, anthraquinone and cardiac 
glycoside while the methanol extract, petroleum ether and ethyl acetate fractions exhibited significant 
(p<0.05) anti-inflammatory activities when compared to the effect of the reference drug Diclofenac 
sodium. 
Keywords:anti-inflammatory, Clerodendrum volubile, macroscopy, microscopy, phytochemical 
screening 
 
 
. 
Introduction 
Clerodendrum volubile P. Beauv (Family; Verbenaceae) also 
known as the white Butterfly leaf is a climbing shrub grown in 
deciduous forests from Senegal to Fernando Po. [1] In folklore 
medicine, the plant is used to treat swellings, oedema, gout, 
arthritis, rheumatism and venereal diseases.  [1]  
 Previous studies have reported the presence of manganese, 
copper, iron, phenols and flavonoids in the leaf of C. volubile [2]  
while, Fleischer et al., 2011 in their study reported the antimicrobial 
and anti-inflammatory activities of the petroleum ether, ethyl 
acetate and ethanol extracts of a species of Clerodendrum, 
Clerodendrum splendens leaves. [3] 
In order to justify the use of Clerodendrum volubile in the folklore 
treatment of inflammation, this study investigates the 
pharmacognostic properties and anti-inflammatory activities of 
Clerodendrum volubile leaf methanol extract and fractions. 
Material and Methods 
Plant Material 
The leaves of Clerodendrum volubile were collected from Agbele 
Community High School, Sagamu, and Ogun State, Nigeria.  The 
plant was authenticated at the Department of Pharmacognosy, 
Olabisi Onabanjo University, and Sagamu where a voucher 
specimen was deposited. 
Extraction 
The leaves were air dried and ground into fine powder.  The 
powdered plant were macerated in 80% methanol for three days, 
filtered and evaporated to dryness under vacuum.  The residue 
was reconstituted in water and successively partitioned with 
petroleum ether and ethyl acetate. 
Chemicals and Instruments 
 Digital camera, Compound microscope, glass and cover slips.  All 
chemicals and reagents used were of analytical grade. 
Phytochemical Studies 
 The Phytochemical screening of the leaf of C. volubile was 
screened for the presence of alkaloids, saponins, flavonoids, 
Tannins and cyanogenetic glycosides using established 
procedures.  [4-5] 
Pharmacognostic Studies 
Macroscopical Features 
 The macroscopical features of the leaves of C. volubile such as 
the odour, colour, size and shape were studied.  [6] 
Microscopy 
The sections for microscopy were prepared by free hand section of 
the leaf which was cleared with chloral hydrate and mounted in 
glycerine. [7-8]  The leaf was viewed by using a compound 
ISSN: 0975-0185 
 
  
This work is licensed under a Creative Commons Attribution 3.0 License.  
Adediwura et al. International Journal of Phytomedicine 4 (3) 414-418 [2012] 
 
PAGE | 415 | 
 
 
 
microscope and pictures of the sections made using a digital 
camera. 
Anti-inflammatory Studies 
Rats of both sexes weighing between 100 – 130gm obtained from 
the animal house of the Department of Veterinary Medicine, 
University of Ibadan were used for this study.  The rats were 
housed and allowed to acclimatize at the animal house of the 
Department of Clinical Pharmacy and Biopharmacy, Faculty of 
Pharmacy, Olabisi Onabanjo University, Sagamu.  The animals 
were maintained at 25oC, fasted for 12h and allowed free access to 
water before the study. Diclofenac sodium (Novartis) was used as 
the reference drug while egg albumin was used to induce 
inflammation by injecting 0.1ml of fresh egg-albumin into the 
subplantar surface of the right hind paw. [9-11] 
The extracts were re-constituted in water and administered orally 
an hour before inducing the inflammation.  The linear 
circumference of the injected paw was measured immediately after 
injection at 0, 30, 60, 90, 120, 150 and 180 minutes.  Increase in 
the linear diameter indicated oedema in the paw. 
% Inhibition 
The percentage inhibition for each treatment was determined by 
using the equation  
% Inhibition of oedema = Mean Control – Mean treatment  
                                           Control 
Statistical Analysis 
The results are presented as mean ± standard deviation and 
percentage decrease (%).  The level of significance of the activities 
of the extract and fractions were determined by the one way 
Analysis of Variance (ANOVA). 
Results and Discussion 
The macroscopic study showed that C. volubile has a simple leaf 
with an entire blade, a reticulate venation, acuminate apex, and a 
cuneate base.  The leaves are green with a bland odour. Table 1  
while the phytochemical analysis of the leaf of C volubile showed 
the presence of flavonoids, tannins, saponins, anthraquinone, 
cardiac glycosides and alkaloids.  Table 2 
The microscopic study of the leaf of C volubile revealed the 
presence of a mixture of unicellular non-glandular and awn-shaped 
uniseriate glandular trichomes with one cell stock and multicellular 
head and an anomocytic type of stomata because the stomata 
 
 
 
 
Table 1:  Macroscopic characteristics of fresh C. volubile leaf. 
 
Colour                           Green 
Odour                            Bland 
Taste                              Slightly bitter 
Type                              Simple 
Margin                           Entire 
Venation                        Reticulate 
Shape                             Lanceolate 
Apex                              Acuminate 
Base                               Cuneate 
Surface                           Smooth 
Texture                           Smooth/soft 
Blade length: Breadth   10cm; 3.2cm 
lacked distinct subsidiary cells Fig1, Fig 2.  A comparison of the 
stomata and hair types are in agreement with previous studies 
which have reported the presence of diacytic, anomocytic, 
anisocytic types of stomata in the members of the family 
verbenaceae.  In addition, a comparison of the microscopic, 
macroscopic and phytochemical analysis of C. volubile with 
another Clerodendrum species (Clerodendrum inerme) has 
reported some similarities. [12-13] 
The anti-inflammatory activity of the leaf extracts of C. volubile was 
established by using a non-steroidal (NSAID) model of fresh 
albumin- induced rat paw oedema.  The model used is known to 
act in a biphasic manner where the initial phase of inflammation (0-
2) is attributed to the release of histamine and kinnins while the 
second phase is sustained by the release of prostaglandins.  [14-
15] 
When the activities of the test extracts were compared to the 
untreated control (water), the methanol extract, ethyl acetate and 
pet ether fractions exhibited an anti-inflammatory effect of 70%, 
73.3% and 70% respectively.  The study revealed that the activity 
exhibited by the methanol, ethyl acetate and pet. ether extracts 
were more pronounced than that exhibited by the reference drug 
(Diclofenac Sodium) at 180 min.   
Though the extracts did not exhibit anti-inflammatory effect until 
after 60 mins.  This therefore confirms that the constituents in the 
methanol, pet ether and ethyl acetate of C volubile acted like most 
clinically effective anti-inflammatory drugs.  [16] 
In all the groups except that administered water only (control), 
maximum inflammation was observed at 60 mins Table 3 
At 150 mins, the anti-inflammatory effect of the Pet. ether and ethyl 
acetate extracts were similar (11.1%) Table 3The maximum anti-
inflammatory effect of 15.6%, 20% and 15.6% were exhibited by 
the methanol, ethyl acetate and pet ether extracts respectively  
 
 
 
 
 
 
Adediwura et al. International Journal of Phytomedicine 4 (3) 414-418 [2012] 
 
PAGE | 416 | 
 
 
 
Table 2 : Phytochemical Screening of Leaf of C. volubile. 
 
                   
Anthraquinone 
Saponins Tannins Cardiac glycoside alkaloids Flavonoids 
Free Combined 
+ ++ ++ ++ ++ ++ ++ 
 
 
 
Table3 : Antiinflamatory Activities of the methanol extract, Pet ether, and Ethyl acetate fractions of Clerodendrum volubile leaf 
 
                 MEAN RAT PAW SIZE (cm) 
     0           30            60           90               120          150       180 (min) 
Water  2.20 ± 
0.21      
2.35  
±0.07 
2.40 
±0.14 
2.75 ±0.21 2.80 
±0.35 
2.90 
±0.56 
3.00 ±0.14 
Petroleum 
ether 
2.40 
±0.14 
2.55  
±0.14 
2.90 
±0.07 
2.60±0.14 
**5.5% 
2.10±0.14 
**25% 
2.00±0.0
0 
**31.0% 
1.90±0.14 
**70% 
Ethyl acetate 2.30± 
0.14 
2.40± 
0.14 
2.70 
±0.14 
2.40± 
0.14** 
12.7% 
2.35 
±0.25 
**16.1% 
2.00± 
0.10 
**31% 
1.80 
±0.00** 
73.3% 
methanol 1.95± 
0.07 
2.25±0.0
7 
3.00±0.14 2.35±0.21**
14.7% 
2.30±0.14 
**17.9% 
2.15±0.2
1 
**25.9% 
1.90±0.14 
**70% 
Diclofenac 
Sodium 
2.35±0.07 
 
2.50±0.1
4 
2.65±0.07 2.45± 0.07 
**10.9% 
2.40±0.00 
**14.3% 
2.25±0.0
7 
**22.4% 
2.25±0.07 
**25% 
P<0.05 when compared to the effect of Diclofenac sodium. 
** % Decrease when compared to the untreated control. 
 
 
Fig 1.   Microscopic features of Clerodendrum volubile leaf 
Adediwura et al. International Journal of Phytomedicine 4 (3) 414-418 [2012] 
 
PAGE | 417 | 
 
 
 
 
 
 
Figure 2: Trichomes and stomata types found in Clerodendrum volubile leaf. 
 
 
when compared to the anti-inflammatory effect of the reference 
drug (Diclofenac Sodium) at 180 min.   
Furthermore, this study revealed that the ethyl acetate fraction of C 
volubile was the most active on a time course curve. There was 
however no significant difference in the anti-inflammatory effects 
exhibited by the extract and fractions (p<0.05) when compared to 
the effect of the reference drug Diclofenac sodium. 
It could therefore be suggested that the active extract and/or 
constituents of C volubile leaf may be exhibiting an inhibitory effect 
on chemical mediators such as prostaglandins. 
This anti-inflammatory effect reported in this study and that 
reported in other Clerodendrum species [3] could suggest that 
Clerodendrum species may be a possible source of novel ant 
inflammatory drugs and this study therefore justifies the use of C. 
volubile as an antiiflammatory agent in folklore medicine. 
 
References  
[1]. Burkill HM. The useful plants of West 
tropical Africa.  Royal Botanic 
Gardens, Kew, 1985; 1:319. 
[2]. Erukainure OL, Oke OV, Ajiboye AJ, 
Okafor OY.  Nutritional qualities and 
Phytochemical constituents of 
Clerodendrum volubile, a tropical 
non-conventional vegetable.  Inter 
Food Res J  2011; 18(4): 1393 – 
1399 
[3]. Fleischer TC, Mensah AY, Oppong 
AB and Mensah MLK, Dickson RA 
and Annan K: Antimicrobial and 
Antiiflammatory activities of 
Clerodendrum splendens leaves.  
Pharmacog Com, 2011; 1: 85 – 89. 
[4]. Harborne JB. Method of extraction 
and isolation In. Phytochemical 
Methods, Chapman and Hall, 
London. Third edition 1998; 60-66. 
[5]. Evans WC. Pharmacognosy, 
Saunders. London. Fifteenth edition 
2009; p585  
[6]. Mukerjee PK. Morphological 
Examinations. In: Quality Control of 
herbal drugs.  Business Horizons 
Pharmaceutical Publishers. 2002; 
132-133. 
[7]. Wallis TE.  A textbook of 
Pharmacognosy. J and A Churchill 
Ltd, London. Third edition 1967 
[8]. Lala PK. Lab Manuals of 
Pharmacognosy.  CSI Publishers 
and Distributors, Calcutta Fifth 
edition, 1993  
Adediwura et al. International Journal of Phytomedicine 4 (3) 414-418 [2012] 
 
PAGE | 418 | 
 
 
 
[9]. Akah PA,  Nwambie A. Evaluation of 
Nigerian traditional medicines. I: 
Plants used for rheumatic 
(inflammatory) disorders. J. 
Ethnopharmacol 1994; 42: 179-182. 
[10]. Winter CA. Risley EA,  Nuss GW.  
Carrageenin-induced edema in hind 
paw of the rats as an assay of anti-
inflammatory drugs. Proc. Soc. Exp. 
Biol. Med., 1962; 111: 544 - 547 
[11]. Hess SM, Milonig RC. Inflammation. 
In: Inflammation Mechanism and 
Control, Lepow, I.H., P.S. Ward 
(Eds.). Academic Press, New York, 
USA, 1972; 1-12. 
[12]. Dinesh K, Pravin VB, Zulfikar AB, 
Jeevan SD, Yogesh SK, Santosh 
SB. Macroscopical and 
Microscopical evaluation of leaves 
of Clerodendrum inerme Gaertn.  
Inter J  Bio  Med sci 2011; 2(1): 404 
– 408. 
[13]. Devi VG, Vijayan C, John A, 
Gopakumark K.  Pharmacognostic 
and Antioxidant studies on 
Clerodendrum inerme and 
identification of ursolic acid as 
marker compound.  Inter J Pharm 
Pharm Sci 2011; 4(2): 145 – 148.  
[14]. Di Rosa M.  Biological properties of 
carrageenan. J. Pharm Pharmacol 
1972; 24: 89-102. 
[15]. Di Rosa, M, Papaimitriou J, 
Willioughy DA. A histopathological 
and Pharmacological analysis of the 
mode of action of non-steroidal and 
anti- inflammatory drugs. J Pathol 
1971; 105: 329-356. 
[16]. Vinegar R, Schreiber W, Hugo R.  
Biphasic development of 
carrageenin oedema in rats. J 
Pharmacol Exp Ther 1969; 166: 96-
103. 
 
 
 
 
 
 


PHARMACOGNOSTIC STUDIES AND ANTIINFLAMMATORY ACTIVITIES OF CLERODENDRUM VOLUBILE P Beauv LEAF

https://www.arjournals.org/index.php/ijpm/article/download/627/506

PHARMACOGNOSTIC STUDIES AND ANTIINFLAMMATORY ACTIVITIES OF CLERODENDRUM VOLUBILE P Beauv LEAF

Abstract

Similar works

Full text

Available Versions

Advanced Research Journals