Pharmaceutical Biology

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Hepatotoxicity of the Methanol Extract of Carica

papaya. (Paw-Paw) Seeds in Wistar Rats

Francis V. Udoh & Paul B. Udoh

To cite this article: Francis V. Udoh & Paul B. Udoh (2005) Hepatotoxicity of the Methanol Extract
of Carica�papaya. (Paw-Paw) Seeds in Wistar Rats, Pharmaceutical Biology, 43:4, 349-352, DOI:
10.1080/13880200590951810

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Pharmaceutical Biology
2005, Vol. 43, No. 4, pp. 349–352
Hepatotoxicity of the Methanol Extract of Carica papaya
(Paw-Paw) Seeds in Wistar Rats
Francis V. Udoh1 and Paul B. Udoh2
1
Department of Pharmacology, College of Medical Sciences, University of Calabar, Nigeria; 2Department of Zoology,
Faculty of Science, University of Calabar, Nigeria
Abstract
Activity of Carica papaya Linn. (commonly known
as paw-paw) seed extract in liver physiology of albino
(Wistar) rats was studied. The MeOH extract of
C. papaya seeds were Soxhlet extracted. Tolerated doses
of C. papaya were estimated in acute toxicity studies and
administered orally, single or repeated doses, for 30 days
to adult male rats weighing between 190 and 200 g, which
were divided into four groups of five rats per group.
Group 1 received 10 mg=kg; group 2 received 50 mg=kg;
group 3 received 200 mg=kg; group 4 received normal
saline (1 ml=rat) as control. Twenty-four hours after
treatments, the animals of all groups were sacrificed
and blood samples collected by heart puncture into cen-
trifugal tubes. The blood samples were allowed to coagu-
late before centrifuged at 400 rpm at 4
C for 15 min to
separate the serum for enzyme assays. A portion of liver
was cut off and fixed in 10% normal saline. The result
showed that C. papaya seed extract treatments caused
elevation of rat serum levels of acid phosphatase
(ACP), alkaline phosphatase (ALP), and aspartate
amino transferase (AST). Also revealed was mild to
severe metaplasia of hepatocytes in a dose-related man-
ner as well as proliferation of Kupfer cells and hepatic
cells cirrhosis. These biochemical and pathological
changes indicated liver cell damage and malfunction.
These results, therefore, suggest that seeds of C. papaya
should be used in herbal medicine with care to avoid
toxicity.
Keywords: Carica papaya, hepatotoxicity, liver sections,
Wistar rats.
Accepted: March 1, 2005
Address correspondence to: Francis V. Udoh, Department of Pharmacology, College of Medical Sciences, University of Calabar,
Calabar, Nigeria. Tel.: 0803-5091976; E-mail: pbudoh@yahoo.com
DOI: 10.1080/13880200590951810
Introduction
Carica papaya Linn. (Caricaceae) is a fruit tree com-
monly cultivated in some parts of southern Nigeria. It
is popularly called paw-paw tree and its fruits are called
paw-paw. The ripe paw-paw fruits are used for preparing
Nigerian salads, sweets, or taken alone with a meal to
enhance digestion. The unripe fruit extract is used for
the treatment of hypertension in Nigerian herbal medi-
cine. The seeds have no nutritive value, but in herbal
medical practice, they have been used to treat a variety
of ailments such as typhoid fever and malaria fever.
Recently, our preliminary investigation into the activity
of the seeds extract of C. papaya on male fertility
revealed antifertility properties (Udoh & Kehinde, 1999).
The phytochemical analysis of the ripe fruit paw-paw
indicated the presence of vitamins A and C (Georges &
Pandelai, 1949). The bark of the tree contains latex,
which is the commercial source of an enzyme used in
the production of digestive medicine, canned meat,
leather tanning, and so forth (Georges & Pandelai, 1949).
The roots of C. papaya have been claimed by herbal-
ists to possess antimicrobial properties against micro-
organisms like gonorrhea, and so forth. In West
Africa, the plant is mainly used as diuretic (root and
leaves), antihelmintic (leaves and seeds), and to treat
bilious conditions (fruit) (Dalziel, 1937). Ripe and unripe
fruits (epicarp, endocarp, and seeds) showed very signifi-
cant antibacterial activity with Staphylococcus aureus,
Bacillus cereus, Escherichia coli, Pseudomonas aerugi-
nosa, and Shigella flexneri (Georges & Pandelai 1949).
# 2005 Taylor & Francis Ltd.
350 F.V. Udoh and P.B. Udoh
Literature searches revealed that few scientific investiga-
tions have been carried out on the pharmacodynamic
properties of C. papaya. However, Nigerian traditional
medical practitioners use the seed extract for treatment
of various types of ailments without considering adverse
side effects.
Based on the above facts, it became necessary to inves-
tigate the action of C. papaya on the liver function of
Wistar rats.
Materials and Methods
Animals
Adult male albino Wistar rats weighing between 190 and
200 g were bought from the Animal House Unit, Depart-
ment of Biological Sciences, University of Calabar,
Nigeria. The animals were housed at controlled tempera-
ture (282
C) with a 12-h dark-light cycle and free access
to rat pellets (Agro Feeds Ltd, Calabar, Nigeria) and
water. The animals were allowed an acclimatization
period of 7 days before treatments.
Extraction
Ripe fruits of Carica papaya were bought from Watt
Market, Calabar, Nigeria, between the months of Octo-
ber and December 1999. They were identified by Frank
Apejoye of the Department of Botany Herbarium, Uni-
versity of Calabar (voucher specimen no. 73). The fruits
were cut open to remove the seeds and were washed clean
to remove debris. The clean seeds were sun-dried for 2
days and then oven-dried at 40
C for 1 week to ensure
complete dryness. The dried seeds were ground into
power with the help of an electric blender (Inoulmex,
France).
The powdered sample (100 g) of C. papaya seeds was
wrapped in a thimble and placed in a 500 cm3 Soxhlet
extractor (M&G Scientific Co., England). The sample
was Soxhlet extracted following standard analytical lab-
oratory method at 60
C in absolute ethanol (450 ml) for
72 h. The extract was evaporated to a paste form at 40
C
for 8 h to produce 11% yield of raw sample of C. papaya.
The powder plant-drug was stored for use in the experi-
ment in a refrigerator.
Extract concentration
A gram weight of C. papaya extract was dissolved in
10 ml of normal saline to give a stock concentration of
100 mg=ml. The doses of the C. papaya extract adminis-
tered were calculated based on kilogram body weight of
rat.
Treatment
Forty adult male Wistar rats weighing between 190 and
200 g were divided into four groups (n ¼ 10). Group 1
received a single dose of 5 ml of normal saline (0.85%
NaCl) orally as control; groups 2, 3, and 4 were treated
with a single dose of extract of C. papaya (10, 50, and
200 mg kg 1 day 1), respectively. Another set of 40 male
rats weighing between 190 and 200 g were grouped into
four groups (n ¼ 10) so that group 1 received 5 ml of nor-
mal saline (0.85 NaCl) orally, daily for 3 days, and
groups 2, 3, and 4 received 1, 50, and 200 mg kg 1 day 1
of C. papaya extract daily for 3 days, respectively. After
7 days post-treatment, the animals of all the groups were
anesthetized with chloroform (Sigma, St. Louis, Mo,
USA) to allow easy collection of cardiac blood sample
for biochemical assay and the animals later sacrificed,
and the liver tissues isolated were fixed in 10% formal-
saline.
Biochemical assays
Animals treated with either normal saline as control or
extract of C. papaya (10, 50, and 200 mg kg 1 day 1)
in single dose or repeated doses for 3 days were anesthe-
tized with chloroform, and blood samples were collected
through heart puncture. The blood samples were centri-
fuged at 4000 rpm at 4
C for 15 min to separate the
serum. The method for measurement of enzyme activity
of acid phosphatase (ACP), alkaline phosphatase
(ALP), and aspartate amino transferase (AST) described
by Reitman and Frankel (1957) were used. Enzyme
activity was measured colorimetrically at 546 nm using
the standard method described by Reitman and Frankel
(1957) and Oyedapo and Araba (2001).
Histopathology
A part of each liver was cut off and prepared for light
microscopy by paraffin embedding, 6-mm sectioning,
and staining with hematoxylin and eosin (H&E). The
tissue specimens were histologically processed for mor-
phological investigations and viewed and photographed
using a light microscope (Olympus, Japan) and camera
(Verorex Company, USA).
Phytochemical analysis
The ethanol extract of C. papaya seeds was phytochemi-
cally screened and found to contain glycosides and poly-
phenol, and some traces of alkaloid, saponins,
flavonoids, reducing compounds, phlobatanins, and
hydroxymethyl anthaquinons (Table 1).
 Hepatotoxicity of Carica papaya 351

Table 1. The photochemical screening of Carica papaya seed
extract.
Serial no. Chemical constituent Alcoholic extract
1 Akaloid
2 Glycosides
3 Saponins
4 Flavonoids
5 Reducing compounds
6 Polyphenol
7 Phlobatanins
8 Hydroxymethyl-anthraquinones
þ, present; þþ , present in excess.
Statistical analysis
Results were expressed as means SEM based on at least
five experiments. Significance was determined by using
Student’s t-test for paired data.
Results
Biochemical assays
Acid phosphatase (ACP)
Oral administration of the methanol extract of C. papaya
to rats in doses of 10, 50, and 200 mg=kg reduced the
concentration of serum ACP with respect to control
(Table 2). In contrast, rats treated with the C. papaya
(50 and 200 mg=kg) caused elevated concentration of
ACP in a dose-related manner (Table 2). Decreased or
increased concentration of the serum ACP was signifi-
cantly different (p < 0.05) compared with the control.
Alkaline phosphatase (ALP)
The methanolic extract of C. papaya (10, 50, and
200 mg=kg) administered to rats orally raised serum con-
centration of ALP in a dose-related manner (Table 2).
The increase in the concentration of the serum ALP
was significantly different compared to control
(p < 0.01).
Aspartate amino transferase (AST)
Serum samples of rats treated with the MeOH extract of
C. papaya (10, 50, and 200 mg=kg) exhibited increased
concentrations of the enzyme AST. The elevation of
þ
þþ
the enzyme concentration was dose-related and signifi-
þ cantly different compared with the control (p < 0.05)
þ (Table 2).
þ
þþ Histopathology
þ
þ The histology of the liver sections of rats treated with
normal saline (control experiments) showed normal cell
morphology (Fig. 1a). Comparatively, the histology of
the sections of livers of rats treated with C. papaya seed
extract (10 mg=kg) showed a mild metaplasia of the
hepatocytes without liver cell damage (Fig. 1d). How-
ever, rats treated with C. papaya, 50 and 200 mg=kg,
revealed severe metaplasia of binucleated cells, prolifer-
ation of Kupfer cells, and hepatic cell cirrhosis that
was significant and more pronounced in the rats treated
with 200 mg=kg (Figs. 1b and c).
Discussion
C. papaya has been employed in Nigerian traditional
medicine for the treatment of a variety of ailments. Liver
is a known major metabolic organ in which most sub-
stances foreign to the body are biologically transformed
or metabolized into nontoxic or more toxic compounds
than the original substance (Bowman & Rand, 1980;
Jie, 1995, Loch et al., 1995). This investigation showed
that the administration of a methanol seed extract of
C. papaya to rats elevated serum levels of ACP and
AST in a dose-related fashion.
Elevated serum levels of ALP and AST signified
liver cell cirrhosis and malfunction (Busik et al., 1993;
Grgurevich et al., 1995; Oyedapo, 1996; Oyedapo &
Araba, 2001). It was reported that an extract of the seed
of Garcinia kola (bitter kola) given to rats caused liver
cell damage (Braide, 1990). Similarly, administration of
C. papaya seed extract to rodents resulted in elevation
of liver transaminases and could cause liver cell lesion

Table 2. Effect of Carica papaya seeds extract on alkaline phosphatase (ALP), acid phosphatase (ACP), and aspartate amino trans-
ferase (AST) serum levels of rats.

Group Experiment Dosage mg kg 1 day 1 ACP IU=L ALP IU=L AST IU=L

A Control 1.0 ml of 0.85% NaCl 5.0  0.2 140  0.4 49  0.1

B Carica papaya 200 7.4  0.2d 250  0.3a 95.7  0.3
C Carica papaya 50 6.7  0.2e 223  0.4b 85  0.2
D Carica papaya 10 4.6  0.1f 205  0.07c 74.0  0.4

Results are means  SEM of 5 experiments.

a
p < 0.01; bp < 0.01; cp < 0.05; dp < 0.05; ep < 0.05; fp < 0.05.

p < 0.01;  p < 0.05;  p < 0.01; versus control by Student’s t-test.
352 F.V. Udoh and P.B. Udoh

Figure 1. Photomicrographs of sections of liver morphology: (a) Control; (b) C. papaya (200 mg kg 1 day 1) treated rat liver (arrows
‘‘A’’ show binucleated cell metaplasia and ‘‘B’’ Kupffer cells proliferation); (c) C. papaya (50 mg kg 1 day 1) treated rat liver [arrows
‘‘A’’ and ‘‘B’’ indicated as for part (b)]; (d) C. papaya (10 mg kg 1 day 1) treated liver morphology (arrows show metaplasia of the
hepatocytes) (H & E, 400)

in the same manner as reported for Garcinia kola seed
extract (Braide, 1990; Cummings & Metcalf, 1995). This
observation, therefore, indicated that the use of the seed
extract of C. papaya in herbal medicine practice without
proper scientific evaluation of its activity is dangerous to
health. Therefore, proper dose regimens should be
designed to avoid toxicity. The photomicrographs of
the liver sections of the rats treated with a high dose of
C. papaya (200 mg=kg) confirmed this fact. However,
the extent of liver cell damage administered at low doses
of C. papaya (10 and 50 mg=kg) was not significant.
Therefore, the seeds of C. papaya should be used in
herbal medicine with caution.
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