EFFECT Oil palm, .Xanthine oxidase, 1.0 Introduction Humans and


Achuba ,F. I .
Department of Biochemistry, Delta State University, PMB 1, Abraka Nigeria. [email protected]

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The toxicity of petroleum hydrocarbon across the
living systems is now a common knowledge among the scientific community. What
is lacking is a mini-scale antidote that can be adopted by the inhabitants of
crude oil producing areas of the world. This was the reason for this study. The
study is comprised forty eight female Wister rats divided into six groups of
eight rats each. The rats were fed as flows: Group 1: (Control). Rats in group
2 and 3 were fed with diets treated with 5.0g and 10.0g oil palm leaf
respectively. Rats in group 4 were fed with crude oil contaminated diet (Crude
oil Control). Rats in groups 5 and 6 were fed with contaminated diet mixed with
ground oil palm leaf (5.0 g and 10.0 g respectively). After three and six months
exposure periods respectively, four rats from each group were sacrificed and
the kidney used to prepare supernatant needed for the determinations of lipid
peroxidation and activities of xanthine oxidase,  superoxide
dismutase  and catalase. The
results show that pretreatment of crude oil contaminated diet with oil palm
leaf tend to restore values of lipid peroxidation,
xanthine oxidase activity, superoxide dismutase activity and catalase activity
close to control values. Thus, it is pertinent to state that there exist
potentials in the use oil palm leaf in the treatment of crude oil toxicity. And
indeed setting a fresh agenda for further serious scientific investigations




Keywords: Crude oil, Kidney, Lipid peroxidation, Oil
palm, .Xanthine oxidase,



 1.0 Introduction

 Humans and animals get exposed to crude oil or
its byproducts when these chemicals are released into the surroundings during
oil exploration activities, equipment failures, corrosion, illegal bunkering,
usage, oil theft and illicit refining 1-3. Crude oil stimulates oxidative
stress in animals 4, 5. Lipid peroxidation, xanthine oxidase superoxide
dismutase (SOD) and catalase activities are part of oxidative stress indices
6. Lipid peroxidation elicits oxidative damage in plants and animals and its
value in conjunction with alterations in the level of antioxidants represent a
measure of oxidative stress. Similarly, the activity of xanthine oxidase is a
defense mechanism as well as measure of oxidative stress in animals 6. Report
has it that the deleterious action of crude oil on the kidney is based on
oxidative stress 7.

 Byproducts of the oil palm tree are important
medicinally. This is because the leaf juice have wound healing property while
the sap is used as laxative 8.This is due to 
 compounds rich in medicinal and antioxidant
properties inherent in oil palm leaf 9, 10. 
The antioxidant action is attributed to the presence of phytochemicals (flavonoid,
tannin and phenols) in the leaves of oil palm tree 11. In fact, oil palm leaf
extract contains more antioxidative phenolic compounds than various green tea
extracts 12. Therefore, oil palm leaf extract is a potential source of
functional food ingredient, based on reports of its health benefit 13 .This
study is aimed at evaluating the protective potentials of oil palm leaf against
crude oil contaminated diet induced nephrotoxicity in rats.

2.0 Materials and methods

The crude oil used for this study was obtained from
Nigeria National Petroleum Corporation (NNPC) Warri, Delta State, Nigeria. The
palm leaf used was obtained from Elaeis guineensis tree in Obiaruku,
Delta state, Nigeria Forty eight (48) female albino wistar rats with weights
ranging from 0.088kg to 0.182 kg obtained from the animal house of Department
of Anatomy, Delta State University, Abraka, Nigeria were used for this study.
The rats were housed in a standard wooden cage made up of wire gauze, net and
solid woods and left to acclimatize for one week on grower’s marsh and tap
water at laboratory temperature of 28o C and
12 hour day/ night regime. After the acclimatization period, the rats
were weighed and grouped.

2.1 Preparation of leaf powder.

leaves were isolated from the stock and sun- dried. The dried leaf was then
ground with domestic kitchen blender into a fine powder and stored in a clean
and sealed plastic container

2.2 Treatment of animals

The forty eight (48) female albino wistar rats were
assigned to six (6) groups according to their weights, with eight rats in each
group. Rats in the control, Group 1 were fed with grower’s marsh only. Rats in
Group 2 were fed with grower’s marsh treated with 5g of powdered palm leaf.
Group 3 rats were fed with grower’s marsh treated 10g of powdered palm leaf.
Group 4 rats were fed with grower’s marsh contaminated with crude oil (4ml per
100g of feed).This concentration of crude oil in diet
was established by a pilot study to be tolerated by the rats over a long period.
Rats in Group 5 were fed grower’s marsh contaminated with crude oil (4ml per
100g of feed) plus 5g of powdered palm fronds. While rats in Group 6 were fed
with crude oil contaminated marsh (4ml per 100g of feed) plus 10g of powdered
palm leaves. The rats in each group were allowed access to clean drinking water
while the experiment lasted. The feeds were prepared fresh daily and stale feed
remnants were discarded regularly. This was done every
morning between the hours of 8 am – 9 am and each group provided with 400 g of
the respective diet. The animals in each group were exposed to their
respective diets for three and six months respectively. The National Institute of health guide for the care and use
of laboratory animals (NIH, 1985) was adopted all through the experiment



2.3 Collection
of samples

After the first exposure period (three months), four rats
from each group were sacrificed and the kidneys were harvested. Five grams (5.0
g) of the kidneys were weighed in chilled conditions and homogenized with 5ml
of normal saline in a mortar. The mixture was diluted
with 45 ml of buffered saline before being centrifuged at 2, 500 rpm and the supernatant was transferred into plastic tubes and
stored at – 4o C in
the refrigerator before used for analysis within forty eight hours. This same
procedure was adopted after six months exposure period.

2.4 Determination of lipid peroxidation and xanthine oxidase activity

The activity of
xanthine oxidase in the kidney of rats was measured using the method of
Bergmeyer et. al. 14, a reaction based on the oxidation of xanthine to uric
acid, a molecule that absorbs light maximally at 290 nm. A unit of activity is
that forming one micromole of uric acid per minute at 25oC. Lipid
peroxidation in the kidney of rats was measured by the thiobarbituric acid
reacting substances TBARS, method of Gutteridge and Wilkins 15.Total
superoxide dismutase activity was assayed using the method of Misra and
Fredorich 16. Catalase was assayed as reported by Rani et al. 17  

2.5Statistical Analysis

of variance (ANOVA) and post Hoc Fisher’s test for multiple comparison
was performed using statistical package for social science (SPSS), version 20  to determine statistical significant
differences between means. P values


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