Continental J.

Pharmaceutical Sciences 4: 35 - 39, 2010 ISSN: 2141 - 4149

© Wilolud Journals, 2010 http://www.wiloludjournal.com

TOXICITY AND HAEMATOLOGY STUDIES OF THE ROOT BARK OF HYMENOCARDIA ACIDA, TUL
(EUPHHORBIACEAE)

P.N. Olotu1, H. Ibrahim2, N Iliyas2 and J.S Gushit3

1
Department of Pharmacognosy, Faculty of Pharmaceutical Sciences, University of Jos, Jos, Nigeria.
2
Department of Pharmacognosy and Drug Development, Faculty of Pharmaceutical Sciences, Ahmadu Bello
University Zaria, Nigeria. 3 Department of Science Laboratory Technology, Faculty of Natural Sciences
University of Jos, Nigeria.

ABSTRACT
The research work covers the toxicities and haematology studies of the root bark of Hymenocardia
acida which is claimed by the Hausa in the Northern Nigeria to be used traditionally for the treatment
of headache, chest-pain, rheumatic pain, toothache, ear pain, migraine and sickle cell crisis. The plant
was safe orally on acute toxicity investigation on mice. There was no observable LD50. The mice
survived even the doses greater than 5000mg/kg and up to 11000mg/kg. The histopathological studies
on chronic administration of the root bark in mice fed with 25% w/w of amended diet showed no
observable organ damage but the mice fed with 50%w/w amended diet showed a focal area of hepatic
necrosis and the globule cells of the intestine were covered with progressive mucin and lymphocyte
proliferations were observed with the spleen. This means that the long term use of the plant in
traditional practice should be cautioned. Furthermore, the body weight, feed and water-intake increase
with the mice fed with the lower concentration of the amended diet (25% w/w) but decreased with the
higher concentration (50% w/w) at P<0.05. There was also a prominent change with the post treated
mice. This result may suggest the use of the plant for nutritional purposes at lower concentration.
Analysis of the blood parameters showed very significant increase with the RBC. This justifies the
claim of the plant in the treatment of anaemia. Analysis of the plant on the remaining blood parameters
was not significant.

KEYWORDS: Acute toxicity, Chronic toxicity, Haematology, Hymenocardia acida and

Euphorbiaceae.

INTRODUCTION
The plant Hymenocardia acida is a tree of about 6 m high, gnarled and twisted with characteristics rough rusty-
red bark, of the wooded savanna throughout the region from Senegal to west Cameroons, and widespread in
tropical Africa. The wood is light brown or pink, darkening to orange, close- grained, with conspicuous annual
rings, and hard. It has been said to be brittle and good only for firewood. This may be reflected in certain Ivorian
names meaning ‘The tree which kills the wife’, i.e. when an unfaithful wife goes to collect firewood, the tree
shatters at her tough and a branch pieces her abdomen. (Bouquet and Debray, 1974).

The Gbaya of the central African Republic recognize the tree as producing good firewood; indeed classified it as
‘woman’s firewood’ being good for the hearth and cooking place, long- lasting while the house wife is about
other chores, yet reviving quickly from sleeping embers, with a hot flame and little smoke (Bouquet, 1969). The
tree is used for house- posts in southern Nigeria, and in Gabon where the wood is made into charcoal for
blacksmith’s work. In Kenya and in Uganda, the wood is known for its hardness, denseness, durability and good
resistance to termite- attack. It is used to make pestles and back- cloth mallets. Charcoal made from the branches
is powdered and rubbed on the head for headache in the soudanian region (Burkill, 1985).

The foliage is browsed a little by cattle in Senegal as a supplementary food and in Nigeria. Leaves and leafy
shoots have a considerable medicinal use. When chewed, they have an acid taste. Leaves are prepared into
infusion for use in Senegal for chest complaints and small- pox and with the roots, for deficiency diseases. A
macerate is given for gripe, and leaf- decoction used as an eye wash (Daziel, 1937). A decoction with honey is
taken in Guinea for biliousness. In Ivory coast- upper Volta, a leaf- decoction is used in baths and draughts as a
febrifuge and leaf- powder is taken as snuff for headache or applied topically for rheumatic pains and toothache,
or for the same purposes, leaves may be pulped with an organic acidic substances such as citron juice or sap of
Piliostigma reticulatum (leguminosae: caesalpinioideae), (Kerharo, 1974).

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Justification
The Hausa tribe in the Northern Nigeria has over the years used the decoction of leaves and the stem bark or
root bark of Hymenocardia acida in the treatment of pain of various categories such as migraine, sickle cell
crisis and menstrual pain (Agishi, 2004). So far, no official work has been done on the toxicity and
haematological studies of the root-bark extracts to establish its safety scientifically.

This study aims at establishing the safety of the crude drug with the view of its future development.

MATERIALS AND METHODS

Plant Collection and Identification:
The root bark of Hymenocardia acida was collected from Kudingi village, Zaria, Nigeria. The plant was
identified in the field using keys and description given in the official books (Woody Plant of West Tropical
Africa). The collection (voucher specimen) was confirmed and authenticated at the Herbarium, Biological
Sciences, Ahmadu Bello University (A.B.U.) Zaria (1010).

Acute Toxicity Studies

The Lorke method (1983) was used for this study. Doses of 500, 1000, 2000, 3000, 4000 and 5000 Mg/Kg body
weight (bw) and 8 mice were used. Doses of 10,000 and 11,000 Mg/Kg bw and 3 mice were used orally.

Chronic Toxicity Studies

The use of Hymenocardia acida in traditional medicine is normally a long term administration and this is why
the aim of this study is to monitor the long term effect of this drug on the animal tissue and organs. Male Swiss
albino mice aged 4-6 weeks were used and these were grouped into 3 of 6 mice each. The control group (1) was
given un-amended diet and water. Whereas the treated (2 & 3) groups were given amended diets of 25%w/w and
50%w/w of the plant root bark respectively for 3 months. The pre and post-treatment water-intake and body
weights of each mouse were daily recorded. At the end of the 3 months, the mice were sacrificed and their
various organ weights recorded. The organs checked for histopathology include the liver, the kidney, the heart,
the lung and the skin. The organs were fixed in 10% buffered neutral formalin for 72h before processing. This
was an attempt to maintain the tissues as it was in the ante- mortem before the post- mortem.

The different tissues were then labelled and allowed to dehydrate in graded series of alcohol in the ascending
order of 70%, 80%, 95% and 100% alcohol after which the tissues were cleared with xylene and impregnated
with paraffin wax, separately embedded for sectioning with rotatory microtome and microtome knife. The
tissues were then sectioned at 6- micro thick and were mounted on a clean and grease free slip and then dried in
an oven. The stained slides were examined with the compound microscope at X40 objectives and the results
were recorded. This was repeated with the other 6 mice fed with 50% w/w amended diet and the control group
(guided by the Department of Pharmacology, Ahmadu Bello University, Zaria). This experiment was repeated
for female albino mice.

Haematology
The experiment was aimed at evaluating the effects of the plant on blood parameters with the view of
investigating the traditional claims of the plant by the Hausas in the treatment of sickle cell anaemia. The
parameters observed were full blood count or differential count for complete blood count (CBC), packed cell
volume (PCV), mean haemoglobin concentration (MHC), total red blood cell count (RBC count), mean
corpuscular volume (MCV), total white blood cell count (WBC count), differential Leucocyte count (LC), and
platelet count.

Swiss albino mice were fed with amended diet containing 25%w/w and 50% w/w of the root bark. After 65 days,
the mice were euthanized and the blood parameters were evaluated using standard methods described by
(Akinloye and Olorede, 2004). The results were compared with animals fed on unamended diet.

RESULTS AND DISCUSSION

Acute toxicity studies
The water extract of Hymenocardia acida when administered orally was found to be safe. No death was
recorded nor was any sign of toxicity observed within 24h of administration even at a higher dose of
11,000mg/kg although LD50 of values greater than 5,000mg/kg have no therapeutic value (Lorke, 1983).

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 P.N. Olotu et al: Continental J. Pharmaceutical Sciences 4: 35 - 39, 2010

Chronic Toxicity Studies

Over 90 days of the experimental studies, the mice fed with 25% w/w amended diet did not show any observable
tissue damage in the lungs, hearts, kidneys, brain intestines and spleens. The control rats also showed no visible
liver damage but the rats fed with 50% w/w amended diet reacted differently. They showed a focal area of
hepatic necrosis and the globule cells of the intestine were covered with progressive mucin and lymphocyte
proliferations were observed with the spleen. But obviously there was no tissue damage of the pancreas, hearts,
lungs, kidneys and brain. This is to say that prolonged (chronic) treatment with this drug in traditional medicine
should be cautioned.

Furthermore, the body weight, feed and water-intake increase with the mice fed with the lower concentration of
the amended diet (25% w/w) but decreased with the higher concentration (50% w/w). There was also a prominent
change with the post treated mice. This result may suggest the use of the plant for nutritional purposes at lower
concentration. The different plates (1, 2, 3 and 4) are shown below.

Haematology studies
Analysis of the blood parameters showed very significant increase with the RBC (Table 1) this justifies the
claim of the plant in the treatment of anaemia. Analysis of the plant on the remaining blood parameters was not
significant (Table 2, 3, 4, 5, 6 & 7).

Plate 1: Fatty degeneration of the Hepatic cells Plate 2: Necrosis of the Hepatic cells

Plate 3: Progressive mucin seen on

Plate 3: Progressive mucin seen on the Plate 4: The section of the spleen showing
globule cells of the Intestine lymphocyte proliferation

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 P.N. Olotu et al: Continental J. Pharmaceutical Sciences 4: 35 - 39, 2010

Table 1: Red blood cells (RBC) Root bark

ANOVA
Source of Variation SS DF MS F P-value F crit
Between Groups 3.754167 2 1.877083 4.147663 0.040437 3.805567
Within Groups 5.883333 13 0.452564
Total 9.6375 15

Table 2: Packed cell volume (PCV) Root bark

ANOVA
Source of Variation SS DF MS F P-value F crit
Between Groups 1.914167 2 0.9570833 1.671574 0.225917 3.805567
Within Groups 7.443333 13 0.5725641
Total 9.3575 15

Table 3: Hb (Root bark)

ANOVA
Source of Variation SS DF MS F P-value F crit
Between Groups 331772.4 2 165886.2 0.807852 0.466986 3.805567
Within Groups 2669449 13 205342.3
Total 3001222 15
Table 4: TP (Root bark)
ANOVA
Source of Variation SS DF MS F P-value F crit
Between Groups 1.914167 2 0.957083 1.671574 0.225917 3.805567
Within Groups 7.443333 13 0.572564
Total 9.3575 15

Table 5: White blood cells (WBC) Root bark

ANOVA
Source of Variation SS DF MS F P-value F crit
Between Groups 18.00417 2 9.002083 0.83108 0.457452 3.805567
Within Groups 140.8133 13 10.83179
Total 158.8175 15

Table 6: Neutrophils (Neu) Root bark

ANOVA
Source of Variation SS DF MS F P-value F crit
Between Groups 21.02083 2 10.51042 0.879155 0.438423 3.805567
Within Groups 155.4167 13 11.95513
Total 176.4375 15
Table 7: Lymphocytes (Lymp) Root bark
ANOVA
Source of Variation SS DF MS F P-value F crit
Between Groups 30.52083 2 15.26042 1.150616 0.346665 3.805567
Within Groups 172.4167 13 13.26282
Total 202.9375 15

CONCLUSION
The toxicity and haematology studies of the root bark of Hymenocardia acida have been carried out
successfully. The results obtained could serve as the tool for establishing the safety of the use of this plant in
traditional medicine and in anaemia.

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REFERENCES
Agishi, E. C., (2004). Etulo, Idoma, Igede, Tiv and Hausa names of Plants. AGITAB Publishers Ltd. Makurdi,
Nigeria. pp.188.

Akinloye, J., & Olorede, M. (2004). Toxicity study of the Leaves of Cochlospermum planchonii. Vol. 13. Pp 23.

Bouquet, A., & Debray, M. (1974): Medicinal plants of the Ivory Coast, Trav. Doc. Orst 32: (Serv. Cent.
Document Orstambondy 93140 France) pp.1

Bouquet, A. (1969). Medicinal plants of the Ivory Coast, Trav. Doc. Orst pp 116.
nd
Burkill, H. M., (1985). The Useful Plants of West Tropical Africa, 2 Edition, Vol.1. Royal Botanic Gardens,
Kew, 960p.

Daziel, J. M., (1937). Useful plants of Tropical West Africa. Crown Agent for Overseas Government and
Administration, London. pp 52-53.

Kerharo, J. (1974). Historic and Ethnopharmacognosic review on the belief and Traditional practices in the
treatment of sleeping sickness in West Africa, Bull Soc. Med. Afri. Noire Lang. FR. 19: (Fac. Med. & Pharm
Dakar Senegal) pp. 400.

Lorke, D.A. (1983). A New Approach to Practical Acute Toxicity Testing. Arch. Toxicology 54:275-287.

Received for Publication: 07/09/2010

Accepted for Publication: 19/10/2010

Corresponding Author:
J.S Gushit
Department of Science Laboratory Technology, Faculty of Natural Sciences University of Jos, Nigeria.
Email: johngushit@yahoo.com

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