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Research Article
Odangowei et al. WORLD JOURNAL OF PHARMACEUTICAL
World Journal of Pharmaceutical and Medical Research
AND MEDICAL RESEARCH ISSN 2455-3301
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EVALUATION OF NATURAL AND ARTIFICIAL RIPENED PINEAPPLE (ANANAS

COSMOSUS L. MERR.) FRUITS ON BIOCHEMICAL PARAMETERS OF WISTAR RATS

Timipa R. Ogoun1 and Odangowei I. Ogidi*2

1
Department of Human Anatomy, Bayelsa Medical University, Yenagoa, Bayelsa State, Nigeria.
2
Department of Biochemistry, Federal Polytechnic Ekowe, Yenagoa, Bayelsa State, Nigeria.

*Corresponding Author: Odangowei I. Ogidi

Department of Biochemistry, Federal Polytechnic Ekowe, Yenagoa, Bayelsa State, Nigeria.

Article Received on 07/06/2022 Article Revised on 27/06/2022 Article Accepted on 17/07/2022

ABSTRACT
Fruits are of high significance worldwide due to their nutritional and commercial values. Likewise, they are
sources of essential nutrients; fortify diet with vitamins and minerals. Artificial ripening may contaminate the fruit
and may cause toxicity. This study was aimed at evaluating natural and artificial ripened Pineapple (Ananas
cosmosus) fruit juice on the Biochemical parameters of Wistar rats. Twenty four (24) adult Wistar rats weighing
between 126.9- 213.3 g was used for this study. The experimental Wistar rats were grouped into three and were
allowed to acclimatize for two weeks at libitum. Five (5) ml/kg of natural and calcium carbide ripened Pineapple
fruit juice were administered orally. At the end of the four weeks feeding period, the rats were sacrificed through
cervical dislocation. Blood was collected by cardiac puncture, using 5ml syringes and 23G needles into blood
sample containers for biochemical analysis using the standard principles of biochemical analysis. The biochemical
parameters evaluated were creatinine, urea, albumin, total protein, transaminases (AST, ALT), alkaline
phosphatase (ALP), total cholesterol, total bilirubin and lactate dehydrogenase and were compared with the control
and natural ripened groups. All mean values of ALT, ALP, Urea, creatinine, Albumin and total cholesterol were
lower when compared with the natural ripened group. The concentrations of AST, total bilirubin, total protein and
lactate dehydrogenase were observed to be higher when compared with the natural ripened group. Statistically, all
biochemical parameters evaluated had significant differences at 95% confidence level (P < 0.05), except creatinine,
albumin and total proteins. This indicates that, consuming calcium carbide ripened pineapple fruits formulated
diets may cause myocardial infarction, hepatic dysfunction and liver and heart diseases, therefore artificial ripening
by the use of calcium carbide method should be discouraged.

KEYWORDS: Artificial, Natural, Pineapple (Ananas cosmosus), Biochemical parameters, CaC2, Wistar rat.

INTRODUCTION chemicals are responsible for different kinds of health

burdens such as weakness, skin disease, kidney failure,
Fruits are of high significance worldwide due to their
cancer, lung failure, dizziness, ulcer, heart diseases.[3]
nutritional and commercial values. Likewise, they are
However, these accelerated ripening processes may also
sources of essential nutrients, fortify diet with vitamins
alter the fruit metabolism and fruit quality due to varying
and minerals, and provide variety to food and make food
physiological processes influenced by multiple biological
appetizing. They further account for a substantial
and environmental factors.
fraction of the world’s agricultural output, and, some
have acquired great economic importance. Fleshy and
Pineapple (Ananas comosus) is a tropical plant with an
juicy fruits are used in producing jams, marmalade, ice
edible fruit; it is the most economically significant plant
creams, yoghurt and juices. In Sub-Saharan Africa,
in the family Bromeliaceae. The pineapple is indigenous
agriculture (fruits production a mainstay) accounts for
to South America, where it has been cultivated for many
about 32% of gross domestic product (GDP).[1]
centuries. The fruit is juicy with excellent flavor and
taste. Pineapple is now considered to be the third most
At present, various ripening agents are being used in the
important fruit crop in world production after banana and
process of fruit ripening. Naturally ripened fruits are
citrus.[4] Pineapple contains different vitamins,
perishable; therefore, various chemicals like ethephon,
antioxidants, a proteolytic enzyme, bromelain which
ethylene, and calcium carbide that are hazardous to
helps in the digestion process by breaking down proteins.
human health are being used to ripen and enhance the
Bromelain has anti-inflammatory, anti-clotting and anti-
fruits' shelf-life. Artificial ripening may contaminate the
cancer properties. Furthermore, it can also interact with
fruit and may cause toxicity.[2] Consequently, these

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 Odangowei et al. World Journal of Pharmaceutical and Medical Research

other medications. Care should be taken when animal house of the Bayelsa Medical University;
consuming fresh pineapple with medications (antibiotics, following the guidance of National Research Council,
anticoagulants, blood thinners, anticonvulsants, Guide for the Care and Use of Laboratory Animals,
barbiturates, benzodiazepines, insomnia drugs and 2011.[9]
tricyclic antidepressants) due to the possibility of certain
side effects associated with it.[5] Administration of samples
LD50 was done using Lorke (1983).[10] Method for
Both natural and chemical ripening procedures are used administration of samples.
for pineapples. However, different ripening procedures
may have different nutrient values and sensory profiles, Group 1: Normal control group of 8 rats (4 males and 4
influencing consumers' choices.[6] The final fruit ripening females) received normal water and feeds only as
stage involves a sequence of biochemical and placebo.
physiological occurrences prime to changes in texture, Group 2: Feeding Group (1) of 8 rats (4 males and 4
color, aroma, and flavor that shape the fruits more females) received 5 ml/kg naturally ripped pineapple
palatable, attractive, and tasty.[7] The rising demand of (Ananas comosus) juice.
fruit safety has inspired researchers about the risk related Group 3: Feeding Group (2) of 8 rats (4 males and 4
to the use of chemically ripened fruits.[8] The present females) received 5ml/kg Calcium Carbide ripened
study was therefore conducted to investigate the possible pineapple (Ananas comosus) juice. The Feeding lasted
changes in biochemical parameters in wistar rats fed on for four weeks.
diets containing natural and artificial (calcium carbide)
ripened pineapple (Ananas comosus) in Yenagoa, Blood sample collection
Nigeria. The animals were observed in their cages for clinical
symptoms daily and at the end of the four weeks
MATERIALS AND METHODS treatment, the rats were sacrificed under chloroform
anesthesia and blood was collected by cardiac puncture,
Fruit and calcium carbide sample collection
using 5 ml syringes and 23G needles into blood sample
Mature unripe pineapple (Ananas comosus) fruits were
containers. The blood was allowed to stand for 2 hours to
plucked off from the pineapple (Ananas comosus) plant
coagulate and was centrifuged for 10 minutes at 2000
in Yenagoa, Bayelsa State. The fruits were divide into
rpm and the supernatant (Serum) carefully collected for
two groups, one group was kept and allowed to rip at
biochemical analysis.
normal room temperature and the second category was
induced to ripe with calcium carbide at the Histology
Biochemical analysis
Laboratory, Bayelsa Medical University, Yenagoa,
Serum levels of aspartate aminotransferase (AST) and
Bayelsa State. Calcium carbide was bought at Swali
alanine aminotransferase (ALT) were determined
Market, Yenagoa, Bayelsa State. 10gram of Calcium
following the principle described by.[11] while the
carbide was placed in a bowl and 5ml of water was used
alkaline phosphates (ALP) were carried out according to
to dissolved it in a closed metal bucket containing 1kg of
the method described by.[12] to assess liver function.
the fruit rapped with black nylon and was allowed for
Renal function was assessed by measuring plasma
two days (48 hours) for ripening. After ripening, sampled
creatinine (CREA) levels and blood urea was assayed
fruits were washed and juiced.
following the method of.[13,14]
Preparation of sample
In order to assess the synthetic function of the liver, total
In this study, 600g of both the naturally ripened and
protein (TP), Total bilirubin and albumin (ALB)
calcium carbide ripened pineapple (Ananas comosus)
concentrations were determined according to the
fruits were peeled separately and blended in an electric
principles based on the Biuret reaction.[15,16] and
blender with 350 ml/1L of distilled water. The juice was
bromocresol green reaction.[17] respectively. Total
filtered with a clean fine sieve and was poured into clean
cholesterol (TC) and Lactate dehydrogenase
bottles and labeled (CaC2 ripened pineapple (Ananas
concentrations were estimated following the method
comosus) fruits juice and naturally ripened pineapple
described by.[18]
(Ananas comosus) juice) then, stored in a refrigerator for
further usage.
Analysis of Data
Data collected from this study was analyzed as Mean ±
Experimental Wistar Rats
Standard Error of Mean (SEM). Significant difference
Twenty four (24) adult Wistar rats (12 male and female
among the groups were determined as P<0.05 by two-
of each sex) weighing between 126.9- 213.3 g was used
way ANOVA, using Statistical Analysis Program for
for this study. The experimental Wistar rats were
Social Sciences (SPSS 22.0 Version).
grouped into three and was allowed to acclimatize for
two weeks (fed with grower mash with clean water) at
libitum then, different dosage of the fruit juice were
administered orally based on their body weight. They
were kept in standard environmental condition in the

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 Odangowei et al. World Journal of Pharmaceutical and Medical Research

RESULTS with the control and natural ripened groups. All mean
values of ALT, ALP, Urea, creatinine, Albumin and total
Biochemical parameters
cholesterol were lower when compared with the natural
Table 1 shows the Body Weight of wistar rats
ripened group. The concentrations of AST, total
administered with natural and CaC2 ripened pineapple
bilirubin, total protein and lactate dehydrogenase were
fruit juice and the control. While Table 2 shows the
observed to be higher when compared with the natural
effects of natural and CaC2 ripened pineapple fruit juice
ripened group. Statistically, all biochemical parameters
on biochemical parameters of wistar rats at 5 ml/kg dose.
evaluated had significant differences at 95% confidence
The biochemical parameters evaluated were creatinine,
level (P < 0.05). Except parameters like creatinine,
urea, albumin, total protein, transaminases (AST, ALT),
albumin and total proteins were not significant at 95%
alkaline phosphatase (ALP), total cholesterol, total
confidence level.
bilirubin and lactate dehydrogenase and were compared

Table 1: Body weight of Wistar rats in grams.

Group Control Natural ripened pineapple fruit juice CaC2 ripened pineapple fruit juice
Mean ±SEM 214.30±10.53 184.53±19.53 174.28±17.35

Table 2: Biochemical Results of natural and CaC2 Ripened Pineapple fruits.

Natural ripened CaC2 ripened
Biochemical parameters Control
pineapple fruit juice pineapple fruit juice
AST [u/l] 50.73±2.08A 66.40±0.20D 67.60±0.80M
F Y
ALT [u/l] 31.10±0.10 49.10±3.30 47.60±1.80A
I S
ALP [u/l] 70.60±2.00 85.80±2.60 85.50±0.70A
Z Z
Creatinine [mg/dl] 0.62±0.02 0.75±0.03 0.73±0.04Z
P T
Urea [mg/dl] 15.50±0.30 24.90±0.50 24.3±1.80R
Total Biluribin [mg/dl] 0.34±0.02S 0.60±0.06X 0.64±0.02H
B B
Albumin [g/dl] 4.50±0.10 4.40±0.20 4.05±0.05B
A A
Total Protein [g/dl] 8.50±0.10 7.80±0.60 7.90±0.30A
W Q
Total Cholesterol [mg/dl] 73.00±3.40 105.25±6.95 95.20±2.40A
X G
Lactate dehydrogenase [u/l] 151.00±2.40 196.60±1.80 199.7±1.90B
Mean±SEM, The Means with Different superscript alphabets in the same row indicates significant difference at 95%
confidence level (p<0.05).

DISCUSSION Increase in bilirubin concentration as seen in the calcium

carbide ripened pineapple group may be suggestive of a
The Biochemical results in this study indicated a slight
hepatobiliary disorder occasioned by the complete or
decrease in serum total protein in the rats fed with
partial blockage of the bile ducts due to increased tension
calcium carbide induced pineapple fruit formulated diet,
on the bile ducts owing probably to the presence of
compared with the control. The reduction in protein may
toxicants or possibly infections which consequently leads
be attributed to increased proteolysis.[19] or probably
to the accumulation of plasma bilirubin [24, 25]. A
reduction in the synthesis of protein which maybe
bilirubin test is important in establishing the underlying
occasioned by the presence of a toxicant which may
factor for jaundice. Hyperbilirubinema may result in the
consequently have resulted into a shift in nitrogen
buildup of bilirubin in the brain which is a risk factor in
metabolism.[20] The observed decrease agrees with the
neurological disorder. Hyperbillirubinaemia is prevalent
information from emerging data suggesting that toxicants
in parenchymal liver disease and biliary obstruction.[26]
such as arsenic and phosphorus present in calcium
In the current study, the highest level of bilirubin was
carbide may leak into the fruit in the course of
noticed in the rats fed calcium carbide ripened pineapple
ripening.[21]
formulated diet which differed significantly from the
control and naturally induced ripening methods. This
The plasma albumin level was observed to decrease in
suggests that calcium carbide increases the level of
the calcium carbide fed group when compared with the
billirubin more than the natural ripening method which
control and the natural ripened fed group. This could be
lends support to earlier report of hepatic dysfunction
adduced that with the induced dosage and the period of
induced by toxicants such as Arsenic found in calcium
feeding calcium carbide does not cause dehydration or
carbide.[27,28]
liver function impairment since the liver is responsible
for the metabolism of protein.[22] The hepatic cells are
Results obtained for creatinine and Urea revealed that no
solely responsible for the synthesis of plasma albumin
significant difference was found in the calcium carbide
which is very vital in the regulation of the blood pressure
induced fed group when compared to the naturally
and the binding and transportation of cellular
induced fed group. Creatinine and urea are analysis
components.[23]

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 Odangowei et al. World Journal of Pharmaceutical and Medical Research

carried out to evaluate kidney function.[29] Creatinine is The decrease in total cholesterol may probably be an
produced from creatine as a waste product of normal indication of slight degenerative changes in the liver.[39]
breakdown of muscle tissue which is then filtered out Meanwhile, there was a significant increase in the levels
through the kidneys without re-absorption and excreted of Lactate dehydrogenase (LDH) in the calcium carbide
in the urine. Blood level of creatinine is elevated when ripened fed group observed when compared with the
this infiltration is deficient, thus creatinine levels are control and natural ripened fed groups which can be lead
used as a test of renal function.[30] Since no significant to heart diseases. These findings were in agreements with
difference was observed for both parameters it implies the studies of Pauline et al.[40]
that the kidney was not yet negatively affected by the
artificial ripening methods. This report agrees with CONCLUSION
earlier report.[31,20] who submitted no significant
The results obtained in this study indicates that all mean
difference in both parameters.
values of ALT, ALP, Urea, creatinine, albumin and total
cholesterol of calcium carbide ripened fruit group were
The marker enzymes of hepatic function showed a varied
lower when compared with the natural ripened fruit
response of the experimental rats. Aspartate transaminase
group. The concentrations of AST, total bilirubin, total
(AST) in the calcium carbide fed group was slightly
protein and lactate dehydrogenase of calcium carbide
elevated when compared with the natural ripened fed and
ripened fruit group were observed to be higher when
control groups and also significant statistically. This
compared with the natural ripened fruit group. This
finding agrees with previous report of Igbinaduwa and
indicates that, consuming calcium carbide ripened
Aikpitanyi-Iduitua,[32] and Gbakon et al.,[20]
pineapple fruits formulated diets may cause myocardial
infaction, hepatic dysfunction and liver and heart
When compared to the natural ripened fed group, there
diseases, therefore artificial ripening by the use of
was a slight decrease in the levels of Alkaline
calcium carbide method should be discouraged.
Phosphatase (ALP) and Alkaline transaminase (ALT)
was observed in the calcium carbide ripened fed group.
ACKNOWLEDGEMENT
ALP are a set of vital metallo enzymes needed for the
hydrolysis of ester bond and the cleaving of phosphorus I want to use this medium to acknowledge Mr. Moses
thereby creating an alkaline environment suitable for Itugha and Preye Sidney O. for their contributions in the
cellular functions.[33] They are abundant in the hepatic laboratories.
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