High Technology Letters ISSN NO : 1006-6748

PHYTOCHEMICAL SCREENING AND ASSESSMENT OF ANTI-

ANAEMIC ACTIVITY OF FICUSCARICA LEAVES IN
PHENYLHYDRAZINE INDUCED ANAEMIC RATS
LAXMAN CH,LAKSHMAN KUMAR CH, PAVANI REDDY M, SUPRIYA N,
VASUDHA B, NERELLA MOUNIKA*
School of Pharmacy, Anurag University, Venkatapur, Ghatkesar, Hyderabad,
Telangana, India.

Corresponding Author:
NERELLAMOUNIKA
Assistant Professor,
Dept. of Pharmacology,
School of Pharmacy, Anurag University,
Venkatapur, Ghatkesar,Medchal,
Hyderabad, Telangana, INDIA – 500088.
ABSTRACT
Nature is the best combinatorial chemist and perhaps has answers to most of the diseases of
mankind. Failure of few synthetic drugs and its possible side effects are the reason for
building interest in many of the researches to seek out help from the ancient healing methods,
that mainly use herbal medicines to give relief. Today herbal remedies are back in repute.
Finding healing powers in plants is an ancient idea. People on all continents have long
applied poultices and imbibed infusions of hundreds, if not thousands, of indigenous plants,
dating back to prehistory. Ficus carica, also popularly known as common fig, its health
management properties have been mentioned in religious books, it is one of the traditional
Mediterranean species belonging to the family of Moraceae. The aim of the present study is
to assess the anti-anaemic activity of ethanolic leaves extract of Ficus carica against
phenylhydrazine induced haemolytic anaemia in rats. Phenylhydrazine (40mg/kg) was
administered intraperitoneally for 2 days to induce anaemia in rats. The animals were divided
in to four groups of 6 animals each. Group I served as normal control, group II as anaemic
control, group III as reference control administered with vitaminB12 and group IV animals
were treated with 200mg/kg, of ethanolic leaves extract of Ficus carica. All the test drugs
were administered once daily for 14 days through oral route. On 15thday blood was
withdrawn, through sinus puncture under phenobarbitone anaesthesia and subjected to the

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estimation of RBC, Hb and percentage haematocrit.Both the ethanolic leaves extract of Ficus
carica and vitamin B12 significantly increased the RBC, Hb and haematocrit levels, which
conclude that Ficus carica leaf extract exhibits anti-anaemic activity. This finding is useful
for further advancements in the fields of food supplements, food additives.Ficus carica has
emerged as a good source of traditional medicine for the treatment of various ailments such
as anaemia, cancer, diabetes, leprosy, liver diseases, paralysis, skin diseases, and ulcers. It is
a promising candidate in pharmaceutical biology for the development/formulation of new
drugs and future clinical uses.
Keywords: Anaemia, anti-anaemic activity, Ficus carica, phenylhydrazine, vitamin B12.

Abbreviations: RBC-Red Blood Cells; CBP-Complete Blood Picture; PHZ-

Phenylhydrazine; Hb-Haemoglobin; WHO-World Health Organization; F. carica-Ficus
carica.

INTRODUCTION
India has an ancient heritage of traditional medicine. The material medica provides a great
deal of information on the folklore practices and traditional aspects of therapeutically
important natural products. Indian traditional medicine is based on various systems including
Ayurveda, Siddha, Unani and Homoeopathy. The evaluation of all these drugs is based on
phytochemical and pharmacological approaches which lead to drug discovery often is
referred to as “natural product screening”. Any part of the plant may contain active
components like bark, leaves, flowers, roots, fruits, seeds, etc.Plants have been used for
thousands of years to flavour and conserve food, to treat health disorders and to prevent
diseases including epidemics. The knowledge of their healing properties has been transmitted
over the centuries within and among human communities. Active compounds produced
during secondary vegetal metabolism are usually responsible for the biological properties of
some plant species used throughout the globe for various purposes, including treatment of
infectious diseases.
Anaemia is a blood disorder that is defined as either red blood cell (RBC) count
below normal or red blood cells which are smaller in size than normal or a level of
haemoglobin below normal. There are over 400 types of anaemia, many of which are rare
but, in all cases, there is lower than normal number of circulating red blood cells. Presently,
more than half of the world’s population will experience some forms of anaemia in their

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High Technology Letters ISSN NO : 1006-6748

lifetime. The various forms of anaemia include iron deficiency anaemia, haemolytic anaemia
(destruction of RBCs), vitamin B12 deficiency anaemia, folic acid deficiency anaemia,
anaemias caused by inherited abnormalities of RBCs such as sickle cell anaemia and
thalassemia and anaemia caused by chronic disease. Anaemia has attained epidemic
proportions worldwide and it impairs normal development in children and constitutes a major
public health problem in young children in the developing countries with wide social and
economic implications. It is a very expensive disorder to manage by use of conventional
drugs. Iron deficiency anaemia is managed by prescribing iron supplements and or a diet of
foods rich in iron. Iron pills are taken together with vitamin C to promote iron absorption.
Iron pills are not taken together with antacids and dairy products since these prevent iron
absorption by the body. Iron tablets may have side effects such as abdominal cramping;
nausea; constipation; and dark, hard stools. The WHO categories anaemia as a dreadful
public health problem (prevalence > 40%) for children of not up to five years in 69 countries
and for pregnant women in 68 countries.
According to National Family Health Survey, the occurrence of anaemia in the
developed world is 71%, in the underdeveloped world it is 84% and the global occurrence is
79%. The major cause of anaemia is linked to low iron levels. However, anaemia exists in
association with other causes, such as malaria, parasitic infection, nutritional deficiencies,
drug toxicity as well as genetic or acquired defect. Anaemia is one of the numerous ailments
claimed to have been successfully treated with plant materials by traditional medicine
practitioners. In China for instance, blood diseases such as malformation of blood circulatory
system, anaemia, varicose veins and haemorrhages have been treated with plant materials.
The genus Ficus (Moraceae) constitutes one of the largest genera of angiosperms with
more than 800 species of trees, epiphytes, and shrubs in the tropical and sub-tropical regions
worldwide. It is one of the 40 genera of the mulberry family, Moraceae. Figs have
considerable cultural importance throughout the tropics, both as objects of worship and for
their many practical uses. This plant also invites attention of the researchers worldwide for its
biological activities. The therapeutic utilities of Ficus carica have been indicated in the
traditional systems of medicine such as Ayurveda, Unani, and Siddha. It has been used to
cure disorders of the endocrine system (diabetes), respiratory system (liver diseases, asthma,
and cough), gastrointestinal tract (ulcer and vomiting), reproductive system (menstruation
pain), and infectious diseases (skin disease, scabies, and gonorrhea).

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The leaves are bright green, single, alternate and large, usually
usually up to 11 ft in length.
They are rough hairy on the upper surface and soft hairy on the underside, deeply lobed with
1-55 sinuses. In receptacles the flowers are seen; arise from the axils of old leaves (Figure 2).
Ficus carica leaves are being used in the
the form of either tea or medicine. It is reported
that Ficus carica leaves have shown beneficial effects in gastrointestinal diseases
diseases, respiratory
diseases,, cardiovascular diseases,
diseases diabetes, skin diseases, ulcers, dysentery
dysentery, and hemorrhoids.
In Western herbal, Nicholas Culpeper (1616–1654)
(1616 1654) wrote: “A syrup made of the leaves is
extremely good for the treatment of cough, lung diseases and it dissolves congealed blood
caused by bruises or falls. F. carica leaves are reported to contain 126 phytoconstituents out
of which polyphenolic compounds are predominant. The phytochemical investigation of
Ficus carica was undertakenn and has led to the identification of over 100 compounds, several
coumarins were isolated from it. Multiple flavonoids have been identified from its, leaves,
stem and roots.
The aim of the present study is to assess the anti-anaemic
anti anaemic activity of ethanolic leaves
extract of Ficus carica against phenylhydrazine induced haemolytic anaemia in rats.

Figure 1: Ficus Caricaplant

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High Technology Letters ISSN NO : 1006-6748

Figure 2: Ficus Caricaleaf

MATERIALS AND METHODS

Ethical approval
This experiment was approved by the Institutional Animal Ethical Committee (IAEC)
(I/IAEC/AGI/001/2021 WR ♀♂), School of Pharmacy, Anurag University, Venkatapur,
Ghatkesar, Medchal, Hyderabad, Telangana.
Study area
The investigation was conducted at Pharmacology Laboratory, Department of Pharmacology,
School of Pharmacy, Anurag University, Venkatapur, Ghatkesar, Medchal, Telangana.
Plant material
The bulk quantity of freshly Ficus carica (fig)) leaves were collected from local vegetable
market, Uppal,, Hyderabad, Telangana.

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High Technology Letters ISSN NO : 1006-6748

Figure 3: Dried leaves powder

Animals
Healthy adult albino rats of Wistar strain of both sexes, weighing about 150
150-200 g were
obtained from the animal house of Anurag University,, Venkatapur, Hyderabad. The rats of
either sex was isolated and housed in separate cages during the experimental
experimental period and kept
at room temperature (24± 20C) with a 12: 12 h light/dark cycle. The animals were fed with a
standard pellet diet and provided water ad libitum. All the procedures and protocols were
reviewed and approved by the Institutional Animal Ethics
Ethics Committee of the School of
Pharmacy, Anurag University.
University

Preparation of extract
The collected leaves were soaked in fresh water for about 15-20
15 20 min to wash out the dirt and
mud particles over their surface. Leaves of Ficus caricawas
was under the shade ai
air dried for 4
weeks and mechanically pounded into fine particles using electronic grinder. About 300g of
the pounded plant materials were weighed and extracted by soxhlation for 72 h with90%
ethanol. The extract was then filtered, evaporated to dryness, and
and stored in capped bottles
inside the refrigerator at 4 0C for further pharmacological study.
study

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Figure 4: Soxhlation
Experimental Design:
Preliminary phytochemical studies
tests of ethanolic extract of leaves of Ficus carica was performed
Preliminary phytochemicaltests
by using standard protocols.
Pharmacological screening
In-vivo anti-anaemic
anaemic activity
Anaemiawasinducedinratsbyintraperitonealadministrationofphenylhydrazine(60mg/kg) daily
for 2 days. Rats that developed anaemia with Haemoglobin concentration lower than 13 g/dl
were recruited for the study.
Following the injections, rats were divided into five groups of six rats each.
 Group-II was anon
anon-anaemic
animal(normalcontrol)whoreceived1ml/kgofnormalsalinesolution.
 Group-II
II was served as anaemic
an control.
 Group-IIIservedasareferencecontrol,received
IIIservedasareferencecontrol,receivedvitaminB12syrup(1ml/rat)
syrup(1ml/rat).
 Group-IV
IV animals received 200mg/kg of ethanolic extract Ficus carica Linn. leaves
through oral administration, by suspending
uspending in saline solution. All the test drug was
administered
istered orally, once daily for 14 days.
days

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Statistical analysis
Data were expressed as mean ± SEM. The data were analysed by using one
one-way analysis of
variance (ANOVA) followed by Dennett’s’ test.P values < 0.05 were considered as
significant.

Figure 5:Collection
Collection of blood sample by retro orbital sinus puncture method

Figure 6:Oral administration of drug samples

RESULT AND DISCUSSION
The present studywasdesignedtoscreentheanti-anaemicactivityofethanolic
anaemicactivityofethanolic
extractofleavesofFicuscaricain
inphenylhydrazineinducedanaemiainrats.Literature
edanaemiainrats.Literature review
conducted on this plant indicated the presence of various active constituents like glycosides,
flavonoids, saponins, triterpenoid, carbohydrates, alkaloids etc (Table 1)
1).Since these
compounds were soluble in ethanol and was
was used as solvent for extraction.

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The anti-anaemic activity of Ficus carica leaf extract was tested on the rats which are
priority induced with anaemia by phenylhydrazine. The experiment was carried out for 14
days of dosing, the blood samples of all subjects were collected and gone through the
CBP.Theresults ofCBPofbothextractandstandardanti-anaemicdrugwascompared and finally
noticed that the extract has good efficiency in treating the anaemia. In phytochemical
screening it is found that it contains rich of chemical constituents which are helpful in
treating anaemia.
Table 1:Preliminary phytochemical screening
Constituents Ethanolic extract

Alkaloids +

Amino acids -

Anthocyanins -

Carbohydrates +

Coumarins -

Fixed oils and -

fats
Flavonones +

Flavones +

Glycosides +

Phenols +

Proteins -

Quinones -

Saponins +

Steroids +

Tannins +

Terpenoids +

Volatile oils -

(+) Present, (-) Absent

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Hematological findings of Hb conc. (g/dL), RBC count (million cells/mm3) and

Table 2:Hematological
PCV (hematocrit or PCV)
PC on day 4 post-PHZ
PHZ treatment
Blood parameter Mean RBC Mean Hb Mean HCT
6
(10 /µL) (g/dL) (%)
Group-I (Control) 5.15 ± 0.04 15.5 ± 0.08 46.3 ± 0.32
Group-II
II (PHZ control) 4.20 ± 0.03 12.61 ± 0.07 37.1 ± 0.51
Group-III (Std control) 4.20 ± 0.05 12.6 ± 0.05 37.27 ± 0.10
Group-IV
IV (Test control) 4.12 ± 0.03 12.51 ± 0.03 36.21 ± 0.06
Values were expressed as mean ± S.D. for six rats in each group, significantly different from Group
Group-II
P<0.05.

Figure 7: Day 4 post PHZ treatment

Hematological findings of Hb conc. (g/dL), RBC count (million cells/mm3) and

Table 3:Hematological
PCV (hematocrit or PCV)
PC on day 14 of treatment
Blood parameter Mean RBC Mean Hb Mean HCT
6
(10 /µL) (g/dL) (%)
Group-II (Control) 5.24 ± 0.02 15.7 ± 0.07 47.43 ± 0.11
Group-II
II (PHZ control) 4.38 ± 0.07 13.11 ± 0.18 39.13 ± 0.74
Group-III (Standard control) 4.80 ± 0.03 14.7 ± 0.03 43.66 ± 0.40
Group-IV
IV (Test control) 4.75 ± 0.02 14.45 ± 0.02 43.33 ± 0.08

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Values were expressed as mean ± S.D. for six rats in each group, significantly different from Group
Group-II
P<0.05.

Figure 8: Day 14 of treatment

emic activity of Ficus carica leaf extract was tested on the rats which
The anti-anaemic
are priorly induced with anaemia
emia by phenylhydrazine.
phenylhydrazine. The experiment was carried out for 14
days of dosing, the blood samples of subjects were collected and gone through the complete
blood picture. The results of CBP of both extract and standard anti-anaemic
anti anaemic drug was
compared and finally noticed that the extract
extract has good efficiency in treating the anaemia. In
the phytochemical screening, it is found that it contains rich of chemical constituents which
are helpful in treating anaemia.
CONCLUSION
Fig is a deciduous tree having wide-range
wide of disease management activities as it is a rich
source of anti-oxidants.
oxidants. Traditionally as well as scientifically various experiments it has been
confirmed that fig has potential role in diseases cure. From this review it can be concluded
that, F. carica has many beneficial biological
biological activities, these facts can be established
clinically by further studies. An extensive literature survey revealed that Ficus carica is a
sacred and important medicinal plant used for the ethnomedicinal treatment of anaemia,
bronchitis, constipation, diabetes, fever (jaundice), hemorrhoids, inflammation, liver
disorders, infectious diseases, and many more throughout the world. The anti
anti-anaemic effect

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produced by the F. caricaleaf may be due to its high content of iron which is present in the
plant. Pharmacological studies carried out on the fresh plant materials, crude extracts, and
isolated components of Ficus carica provide an experimental support for its numerous
traditional uses.
The ethanolic leaf extract of the Ficus carica Linn exhibits anti-anaemic activity
against phenylhydrazine induced anaemia in rats.It also shows good anti-oxidant activity as
well.Thebotheffects producedbyFicuscaricaleafmaybeduetoitshighcontent of phenols, sterols,
iron and other chemical components.Further investigations should be conducted to isolate
and characterizethe active components of this plant.Further studies on quantitative estimation
of phytoconstituents along with in-vivoevaluations are necessary for elucidating the exact
mechanism of action of the observed anti-anaemic effect.

Declarations

Acknowledgement

The authors wish to thank the management of School of Pharmacy, Anurag University,
Venkatapur, Ghatkesar, Telangana, India for providing necessary equipment for research,
praiseworthy inspiration, constant encouragement, facilities and support.

Author contributions

All authors contributed to experimental work, data collection, drafting or revising the article,
gave final approval of the version to be published, and agree to be accountable for all aspects
of the work.

Funding statement

This research did not receive any specific grant from funding agencies in the public,
commercial, or not-for-profit sectors.

Competing interest statement

All authors declare that there is no conflict of interests regarding publication of this paper.

Additional information

No additional information is available for this paper.

Financial support and Sponsorship

None.

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REFFERNCES
1. Ayoub L, Hassan F, Hamid S, Abdelhamid Z, Souad A. Phytochemical screening,
antioxidant activity and inhibitory potential of Ficus Carica and Olea europaea
leaves. Bioinformation. 2019;15(3):226.
2. Didangelos T, Karlafti E, Kotzakioulafi E, Margariti E, Giannoulaki P, Batanis G,
Tesfaye S, Kantartzis K. Vitamin B12 Supplementation in Diabetic Neuropathy: A 1-
Year, Randomized, Double-Blind, Placebo-Controlled Trial. Nutrients.
2021;13(2):395.
3. Akorede GJ, Ambali SF, Olatunji AO, Shittu M, Aremu A, Basiru A, Biobaku KT,
Ahmed AO, Ameen SA. Protective effect of vitamin C on chronic carbamazepine-
induced reproductive toxicity in male Wistar rats. Toxicology reports. 2020; 7:269-
76.
4. Silva FR, Matias TM, Souza LI, Matos-Rocha TJ, Fonseca SA, Mousinho KC, Santos
AF.Phytochemical screening and in vitro antibacterial, antifungal, antioxidant and
antitumor activities of the red propolis Alagoas. Brazilian Journal of Biology.
2019;79(3):452-459.
5. Sathiya R, Krishnaraj K, Muralidharan S, Muruganantham N. A simple and validated
HPTLC method of evaluation for quetiapine fumarate in oral solid dosage form.
Eurasian Journal of Analytical Chemistry. 2010 Mar 10;5(3):246-53.
6. Morales-Gutierrez J, Díaz-Cortés S, Montoya-Giraldo MA, Zuluaga AF. Toxicity
induced by multiple high doses of vitamin B12 during pernicious anaemia treatment:
a case report. ClinicalToxicology. 2020 Feb 1;58(2):129.
7. Ramesh C, Lopamudra P; Prevalence of Iron Deficiency Anaemia in India: Results
from a Large Nationwide Survey. Journal of Population and Social Studies, 2010;
19(1): 59-80
8. Roque M, D’Anna C, Gatti C, Veuthey T; Haematological and Morphological
analysis of erythropoietic regenerative responses in Phenylhydrazine induced
haemolyticanaemia in mice. Scandinavian Journal of Laboratory Animal Science,
2008; 35: 181 – 190.
9. Kambou SP, Bleyere MN, Attemene SDD, Tiahou GG, Dembele A, Sess DE.
Antianaemic effect of spirulina in rabbits (Oryctolagus cuniculus), a made and used
food supplement in Côte d’Ivoire. Scholars Academic J Biosci 2015;3(9):725-732

Volume 28, Issue 8, 2022 29 http://www.gjstx-e.cn/

High Technology Letters ISSN NO : 1006-6748

10. NY Gond, SS Khadabadi, Hepatoprotective activity of Ficus carica leaf extract on

rifampicin induced hepatic damage in rats. Indian journal of pharmaceutical sciences,
2008,70 (3): 364-366.
11. Devi PrasunaKalluri, Mohammed Sayeed, Vasudha Bakshi, Narender Boggula.
Phytochemical Screening and Anti-Inflammatory Activity of Moringa oleifera Pods-
An In Vivo Design. The Pharma Innovation. 2018; 7(7): 673-679.
12. Solomon A, Golubowicz S, Yablowicz Z, Grossman S, Bergman M, Gottlieb HE,
Altman A, Kerem Z, Flaishman MA. Antioxidant activities and anthocyanin content
of fresh fruits of common fig (Ficus Carica L.). J Agric Food Chem. 2006; 54:7717.
13. Yancheva SD, Golubowicz S, Yablowicz Z, Perl A, Flaishman MA. Efficient
agrobacterium-mediated transformation and recovery of transgenic fig (Ficus
carica L.) plants. Plant Science. 2005;168(6):1433–1441.
14. Smitha Surendran, Syed Mohammed BasheeruddinAsdaq, PrapullaPutta, Mounika
Nerella, Narender Boggula. Anti-Obesity Screening of Figs (Ficus Carica) In
Animals Fed on Atherogenic and Cafeteria Diet. Journal for Innovative Development
in Pharmaceutical and Technical Science. 2020; 3(6):1-10.
15. Desna Amelliya, Helmi Arifin, WidyaKardela. A Review: Plants That Have Activities
as Anti-Anemia. IOSR Journal of Pharmacy and Biological Sciences (IOSR-JPBS).
2021; 16(1):48-55.
16. Baskaran K, et al. Anti-Anemic Activity of Ethanolic Leaf Extract of
Kedrostisfoetidissima in Phenylhydrazine Induced Anemic Rats. Biosci. 2016;
4(8):681-683.
17. Joshi Ankur, Soni Priyanka, Vyas Narendra, Khan Javed, Malviya Sapna and Kharia
Anil leaf extract of Aegle marmelos in Phenylhydrazine induced anemic rats.
International Journal of Current Research. 2017; 9(04):48928-48931.
18. Narender Boggula, Swetha Reddy Naram Reddy, Teja Sri Alla, Azmath Farhana,
JainendraBattineni, Vasudha Bakshi. Phytochemical evaluation and in vitroanti
bacterial activity of dried seeds of Abrusprecatorius. International Journal of
Pharmaceutical Sciences Review and Research. 2017;44(1):101-107.

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