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ISSN Online: 2374-6866
SOJ Pharmacy & Pharmaceutical Sciences
Research Article
Open Access
Toxicity Evaluation of the Extract and Fraction of
Chrysophyllum Albidum Seed Cotyledons in Rats
Adedoyin Akinmayowa Shobo1, Michael Oluwatoyin Daniyan1*, Gbola Olayiwola2,
Thomas Oyebode Idowu3, Abiodun Oguntuga Ogundaini3 and Saburi Adejimi Adesanya4
1
2
Department of Pharmacology, Faculty of Pharmacy, Obafemi A wolowo University, Ile-Ife, Nigeria.
Department of Clinical Pharmacy and Pharmacy Administration, Faculty of Pharmacy, Obafemi Awolowo University, Ile-Ife, Nigeria.
3
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Obafemi Awolowo University, Ile-Ife, Nigeria.
4
Department of Pharmacognosy, Faculty of Pharmacy, Obafemi Awolowo University, Ile-Ife, Nigeria.
Received: January 11, 2019; Accepted: February 15, 2019; Published: February 20, 2019
*Corresponding author: Michael Oluwatoyin Daniyan, PhD, Senior Lecturer, Department of Pharmacology, Faculty of Pharmacy, Obafemi Awolowo University, Ile-Ife, 220005, Osun State, Nigeria. Email: toyinpharm@gmail.com, mdaniyan@oauife.edu.ng
Abstract
The seed of Chrysophyllum albidum (G.Don) Sapotaceae, is widely
employed for its economic and medicinal values, necessitating the need
to establish its safety. Therefore, the toxicity profiles of the methanol
extract and butanol fraction of C. albidum seed cotyledons following
acute and 28-day repeated dosing was investigated in rats using
OECD Test Guidelines 420 and 407. The median lethal dose (LD50)
of the extract and fraction were 760 and 200 mg/kg respectively,
and both effect varying degree of significant changes at p < 0.05 in
organ weights, organ-brain weight ratio, hematological indices (WBC,
RBC, Hb, HCT, MCV, MCH, MCHC), and biochemical indices (AST, ALT,
creatinine, direct and total bilirubin) as well as observed pathological
changes following acute and repeated dose administration. The results
showed potential to cause mild to moderate toxicity, especially more
with butanol fraction, and at higher doses. The recovery data showed a
potential for recovery from toxic effects, but also provided a reflection
of delayed toxicity. Hence, while taking advantage of the many
medicinal and economic potentials of C. albidum seed cotyledons, its
potential toxic effects need to be considered.
Keywords: Medicinal plants; Toxicity profiles; Acute toxicity;
Repeated dose toxicity; OECD;
Abbreviations
OECD, Organization for Economic Co-operation and
Development; TG, Test Guidelines; ME and BF, Methanol extract
and Butanol fraction of C. albidum seed cotyledons respectively;
FOB, Functional observatory batteries; FWR, Female Wistar rats;
ALT, Alanine aminotransferase; AST, Aspartate aminotransferase;
WBC, White blood cell; RBC, Red blood cell; Hb, Hemoglobin
concentration; HCT, Hematocrit; MCV, Mean corpuscular volume;
MCH, mean corpuscular hemoglobin; MCHC, Mean corpuscular
hemoglobin concentration.
Introduction
Medicinal plants have been employed in the treatment and
management of many diseases [1,2], and there are increasing
global calls, especially in Africa, to promote and integrate
Symbiosis Group
traditional medical practices into their health system [3]. Cost
effectiveness, ease of accessibility, wider cultural acceptability,
source of raw materials as well as potential chemical candidates
for drug discovery are some of the factors responsible for upsurge
of interest in medicinal plants [1]. However, the extensive and
indiscriminate uses of these plant-based medicines among other
concerns, makes the evaluation of their toxicity imperative [4].
Among the medicinal plants that are widely consumed and
known for therapeutic properties is the Chrysophyllum albidum
(G.Don) Sapotaceae (White Star Apple) [5]. The tree plant is
popularly referred to as “agbalumo” (South-western Nigeria),
“udara” (South-eastern Nigeria), agwaluma (Northern Nigerian)
[2,6]. It is a dominant canopy tree of lowland mixed rain forest
widely distributed throughout the tropical Central, East and
West Africa regions [2,6]. The oil of C. albidum seed cotyledons is
employed in soap and candle production and as lubricant, while
the stem-bark is used as remedy for malaria, sleeping sickness,
and yellow fever [2,7]. The leaves are used as emollients and for
the treatment of skin eruptions, diarrhoea and stomach ache [2,6].
In addition, the C. albidum seed cotyledons have been reported to
have a number of folkloric applications, including wound healing,
treatment of oligospermia, amenorrhea, certain dermatological
and vaginal infections, intestinal worms and hemorrhoids [2,8].
Previous investigation of the chemical constituents of C. albidum
showed that the stem-bark contains stigmasterol, epicatechin,
epigallocatechin and procyanidin B5 [9]. Phytochemical studies
of the crude and methanol extracts of seed cotyledons revealed
the presence of saponin, alkaloid, tannin, flavonoid, sterol and
anthraquinone [10]. Earlier, bioassay-guided fractionation of
the methanol extract of the C. albidum seed cotyledons led to the
isolation of eleagnine, tetrahydro-2-methylharman and skatole
[11]. Like many herbal products containing β-carbolines, the
presence of β-carboline alkaloids in C. albidum seed cotyledons
suggests its potential usefulness in the treatment of cancer,
neurological disorders, malaria, jaundice and asthma [12,13].
The known pharmacological activities of C. albidum seed
*Corresponding author email: toyinpharm@gmail.com, mdaniyan@oauife.edu.ng
Toxicity Evaluation of the Extract and Fraction of Chrysophyllum Albidum
Seed Cotyledons in Rats
cotyledons, attributable to the presence of eleagnine, are antioxidant, anti-inflammatory, anti-nociceptive and anti-microbial
activities [11, 14, 15, 16]. Also reported are the biochemical
effect and membrane stabilization potential of eleagnine [17] and
anti-hyperglycemic and hypolipidemic properties of the ethanol
extract of C. albidum seed cotyledons [18].
However, in spite of these plethoras of pharmacologic
and economic potentials, the toxicity profile remains largely
unexplored. In this study, we report the evaluation of the toxicity
profile of the extract and fraction of C. albidum seed cotyledons.
Materials and Methods
Plant Material: Collection, extraction and fraction
The fresh fruits of C. albidum were bought from Sabo
fruit market at Ile-Ife, Osun State, Nigeria. It was identified
and authenticated by Mr. A.T. Oladele of the Department of
Pharmacognosy, Faculty of Pharmacy, Obafemi Awolowo
University (OAU), Ile-Ife, Osun State, Nigeria, where the
herbarium specimen, with a voucher number FPH/S/001, was
deposited. The plant name was also checked with http://www.
theplantlist.org for confirmation. The seeds were separated from
the fresh fruits, de-shelled to obtain the white cotyledons, and
air-dried at room temperature. The air-dried cotyledons (500
g) were subsequently pulverized using mortar and pestle, and
extracted with 2.5 litres of 100 % methanol three times at room
temperature for 72 hours. The pooled extract was concentrated
in a rotary evaporator to obtain the crude methanol extract
(68 g). Forty grams (40 g) of the crude extract was dissolved in
distilled water and successively partitioned with ethyl acetate
and butanol. The resultant ethyl acetate, butanol and aqueous
fractions were concentrated to dryness in vacuo.
Care and use of experimental Animals
Experiments were performed using nulliparous Female
Wistar rats (FWRs), weighing 130 – 150 g, and bred locally
in the animal holdings of the Department of Pharmacology,
Faculty of Pharmacy, OAU, Ile-Ife, Nigeria. FWRs were housed
in standard plastic cages, exposed to natural room temperature
and lighting conditions, and allowed one week acclimatization
with free access to standard laboratory pellets (Grand Cereals,
United African Company Plc, Nigeria) and water ad libitum. The
procedure for the care and use of animals was in strict compliance
with the recommendations in the “Guide for the Care and Use of
Laboratory Animals – Eighth Edition” of the National Research
Council of the National Academies, USA [19]. The protocol was
approved by the Committee on the care and use of laboratory
animals, Obafemi Awolowo University, Ile-Ife, Nigeria (Protocol
number PHP12/13/H/0601).
Median lethal dose (LD50) determination and Sighting
study
Median lethal dose (LD50) values of ME and BF were
determined using Lorke’s method [20] and Hodge and Sterner
scale [21] was used in the categorization of the degree of toxicity.
A preliminary sighting study [22] using functional observatory
Copyright:
© 2019 Daniyan MO, et al.
battery (FOB) [23] was conducted to determine the humane
endpoint criteria and for the selection of the appropriate doses
for the main study. Test agents were administered orally. FOB
used in this study consists of twenty functional and behavioural
activities, including behavioral changes, motor activity, sensory
reflex, coordination, respiratory distress, vocalisation, oculonasal
discharge, fear and death. Using LD50 values as guide, the starting
dose for the sighting study, selected from the Organisation for
Economic, Co-operation and Development (OECD) Test Guideline
(TG) 420 [22] fixed dose levels of 5, 50, 300 and 2000 mg/kg,
was fixed at 50 mg/kg for BF and 300 mg/kg for ME. Based on
the resultant evident or lack of evident toxicity / mortality,
subsequent dose levels were selected from the OECD fixed dose
levels, including a control. Each dose level uses one (1) FWR with
a period of 24 hours in-between dosing. Where death occurs, a
confirmatory test with a second FWR was conducted. FOB were
monitored continuously for the first 30 minutes, then every 30
minutes for 4 hours, and thereafter at regular interval for 24
hours and daily for a total of 14 days.
Experimental design
The experiment was divided into two test phases: Acute
and repeated dose toxicity studies. FWRs were fasted overnight
before the start of the studies. Test agents were administered
orally.
The acute toxicity study was conducted using the OECD TG
420 [22], with 35 FWRs, randomly allotted to 7 groups of 5 rats
each, namely, control group 1, ME treated groups 2, 3, 4 and BF
treated groups 5, 6, 7. Group 1 receives single oral dose of normal
saline (0.9 % NaCl w/v). Groups 2, 3 and 4 were administered
single oral doses of 150, 300, and 600 mg/kg body weight ME
respectively. Groups 5, 6 and 7 were treated orally with single
doses of 40, 80, 160 mg/kg body weight BF respectively. FWR
body weights were taken before dose administration, and at least
twice weekly for two weeks. FOBS were monitored as described
above.
The repeated dose toxicity test was conducted in accordance
with the OECD TG 407 [24]. Fifty FWRs were used, and were
randomly divided to 5 groups of 10 rats each for control (group
1), ME treated groups 2 and 3 and BF treated groups 4 and 5.
Group 1 received normal saline (0.9 % NaCl w/v), groups 2 and
3 received 100 and 300 mg/kg body weight of ME respectively,
while groups 4 and 5 were given 50 and 150 mg/kg body weight
BF respectively. All doses were administered once daily for 28
days and FWRs body weights were taken periodically. On day 28,
the control and surviving members of each treated groups 2, 3, 4
and 5, were randomly divided into two equal Sets: Toxicity and
Recovery. Recovery set were allowed a further 21 days of nondosing recovery period. Following each daily administration, FOB
were monitored continuously for the first 30 minutes, then once
every 30 minutes for 4 hours.
In all the experiments, doses were prepared using
physiological saline (0.9 % NaCl w/v) and volume of administered
doses was not more than 5 ml / kg body weight. Furthermore, on
day 14 for acute toxicity test, and days 29 and 49 for repeated
Citation: Shobo AA, Daniyan MO, Olayiwola G, et al. (2019) Toxicity Evaluation of the Extract and Fraction of Chrysophyllum
Albidum Seed Cotyledons in Rats. SOJ Pharm Sci, 6(1) 1-12. DOI: 10.15226/2374-6866/6/1/00194
Page 2 of 12
Copyright:
© 2019 Daniyan MO, et al.
Toxicity Evaluation of the Extract and Fraction of Chrysophyllum Albidum
Seed Cotyledons in Rats
dose Toxicity and Recovery sets respectively, the surviving rats
were euthanized using cervical dislocation, and all efforts were
made to minimize suffering. Blood samples were collected by
cardiac puncture into EDTA sample bottles and processed for
hematological and biochemical assays, while organs samples
(brain, liver and kidney) were isolated and weighed. Liver and
kidney were then processed for histopathological analysis.
Statistical analysis
Data were expressed as mean ± standard error of mean (SEM),
and analyzed using Student’s t – test and / or one – way analysis
of variance (ANOVA) followed by Dunnett’s post hoc test using
Graph Pad Prism version 5.01 (Graph Pad software, San Diego,
California, U.S.A). The level of significance was set at p < 0.05.
Results and Discussion
Biochemical assays
Blood samples in EDTA tubes were centrifuged at 3000 rpm
for 5 minutes to obtain the plasma for biochemical assays. Alanine
aminotransferase (ALT), aspartate aminotransferase (AST),
bilirubin (total and direct) and creatinine, were assayed using
standard biochemical kits as per manufacturer’s instructions
(Randox Laboratories Limited, Crumlin, County Antrim, BT294QY,
United Kingdom).
Hematological assays
The whole blood samples were subjected to hematological
analysis using Mindray BC 2800 Haematology Auto-Analyzer
[25]. WBC, RBC, Hb, HCT, MCV, MCH, MCHC were analyzed.
Histopathological examination
The liver and the two kidneys were fixed in 10% buffered
formalin and dehydrated using increasing concentrations of
isopropyl alcohol (80 - 100%). The organs were embedded in
paraffin, and sectioned at 5 µm thickness using a Leica rotary
microtome (Bright B5143 Huntington, England). The sections
were subjected to routine hematoxylin–eosin (HE) staining,
involving deparaffinization, hydration, staining, rinsing and
clearing in xylene in line with standard procedures [26,27]. Slides
were viewed under light microscope with photomicrographs
taken with a Leica DM750 Camera Microscope (X 400).
Medicinal plants remain an indispensable alternative and
complementary pharmacotherapy in the management of many
diseases, necessitating the need to establish their safety [1]. In
this report, we have evaluated the toxicity of C. albidum seed
cotyledons methanol extract and butanol fraction using acute
and repeated dose toxicity models with a view to ascertain their
safety and the possibility of reversibility or persistence of their
toxic effects. Butanol fraction was selected based on our earlier
work that found eleaginine, its essential active phytochemicals,
in butanol fraction following TLC derivatization of the three
fractions using Draggendorff’s reagent [11,17].
Median lethal dose (LD50) and humane endpoint
criteria
The estimated oral LD50 of the ME (760 mg/kg) and BF
(200 mg/kg) provided an indication that they could be slightly
and moderately toxic respectively [21]. Also, the sighting studies
revealed that loss of righting reflex and respiratory distress are
the obvious FOB observed in all rats that died (Table 1), and are
therefore used as humane endpoint criteria in subsequent acute
and repeated dose toxicity studies.
Table 1: Summary of cage side observations following single and repeated oral dose administrations of Methanol extract and Butanol fraction of C.
albidum cotyledon
Single dose oral administration (x/5)
Signs of toxicity
Methanol Extract
(mg/kg)
Control
Repeated dose oral administration (x/10)
Butanol Fraction (mg/
kg)
Methanol Extract
Control
150
300
600
40
80
160
(mg/kg)
Butanol Fraction
(mg/kg)
100
300
50
150
Piloerection*
0/5
0/5
0/5
2/5
0/5
1/5
1/5
0/10
0/10
2/10
2/10
1/10
Reaction to handling
0/5
0/5
0/5
0/5
0/5
1/5
0/5
0/10
0/10
3/10
0/10
2/10
Palpebral closure
0/5
0/5
0/5
0/5
0/5
0/5
0/5
0/10
0/10
5/10
0/10
0/10
Eye colour
0/5
0/5
0/5
0/5
0/5
0/5
0/5
0/10
0/10
0/10
0/10
0/10
Lacrimation
0/5
0/5
0/5
0/5
0/5
0/5
0/5
0/10
0/10
0/10
0/10
0/10
Gait
0/5
0/5
0/5
0/5
0/5
1/5
0/5
0/10
0/10
3/10
0/10
4/10
Sedation
0/5
0/5
0/5
0/5
0/5
0/5
0/5
0/10
0/10
0/10
0/10
0/10
Skin colour
0/5
0/5
0/5
0/5
0/5
0/5
0/5
0/10
0/10
0/10
0/10
0/10
Respiration distress#
0/5
0/5
0/5
2/5
0/5
0/5
1/5
0/10
0/10
2/10
1/10
4/10
Tremor
0/5
0/5
0/5
0/5
0/5
0/5
0/5
0/10
0/10
0/10
0/10
0/10
Convulsion
0/5
0/5
0/5
0/5
0/5
0/5
0/5
0/10
0/10
0/10
0/10
0/10
Defecation
0/5
0/5
0/5
0/5
0/5
0/5
0/5
0/10
0/10
0/10
0/10
0/10
Citation: Shobo AA, Daniyan MO, Olayiwola G, et al. (2019) Toxicity Evaluation of the Extract and Fraction of Chrysophyllum
Albidum Seed Cotyledons in Rats. SOJ Pharm Sci, 6(1) 1-12. DOI: 10.15226/2374-6866/6/1/00194
Page 3 of 12
Copyright:
© 2019 Daniyan MO, et al.
Toxicity Evaluation of the Extract and Fraction of Chrysophyllum Albidum
Seed Cotyledons in Rats
0/5
0/5
0/5
2/5
0/5
1/5
2/5
0/10
0/10
4/10
4/10
5/10
0/5
0/5
0/5
2/5
0/5
0/5
1/5
0/10
0/10
2/10
1/10
4/10
0/5
0/5
0/5
2/5
1/5
1/5
2/5
0/10
8/10
10/10
6/10
8/10
Hypokinesia*
0/5
0/5
1/5
2/5
0/5
2/5
3/5
0/10
0/10
9/10
3/10
6/10
Tail elevation
0/5
0/5
0/5
0/5
0/5
0/5
0/5
0/10
0/10
0/10
0/10
0/10
Inappetence*
0/5
0/5
1/5
2/5
0/5
2/5
4/5
0/10
0/10
3/10
1/10
4/10
Fear
0/5
0/5
0/5
0/5
0/5
0/5
0/5
0/10
0/10
0/10
0/10
0/10
Death
0/5
0/5
0/5
2/5
0/5
0/5
1/5
0/10
0/10
2/10
1/10
4/10
Vocalisation*
Loss of righting
reflex#
Oculonasal
discharge*
Functional observational battery (FOB) in toxicity
studies
The acute toxicity study shows no clear signs of intoxication
(including mortality) aside general malaise such as lethargy,
transient hypoactivity and inappetence in the FWR administered,
except at 600 mg/kg methanol extract (ME) where 40% mortality
was observed and 20% mortality at 160 mg/kg butanol fraction
(BF) (Table 1). On the other hand, repeated dose toxicity study
showed 20% mortality at 300 mg/kg ME, and 10% and 40 %
mortalities at 50 mg/kg and 150 mg/kg BF respectively (Table
1). It should be noted that FOB [23], revealed that while the
behavioural alterations during the course of the acute and 28
days repeated administration are similar, they also correlated
well with the rate of mortality, indicating that BF could be more
toxic than ME (Table 1). It is proposed that the observed toxicity
may be connected with the reported depressant or inhibitory
effect of the test agents on the central nervous system (CNS) [17].
Toxicological assessment of C. albidum seed cotyledons
extract and fraction
Effect on body weight, organ weights and organ-brain
weight ratio
Available evidence has shown that body weight gain, organ
weights and organ - brain weight ratio are important and sensitive
indices of toxic effects [28]. Essentially, organ – brain weight ratio
is a more relevant index for toxicity in cases where significant
variations in body weight is inevitable, as test materials that alter
body weight generally do not alter brain weight [28]. In the acute
toxicity study, though significant differences (p < 0.05) in relative
body weight were observed in all treated FWRs at all tested
doses when compared to control (Table 2), lack of significant
changes in relative organ weights and organ–brain weight ratio
(Table 4) suggest that any potential toxicity of the test agents
may be temporary. On the other hand, following repeated oral
administration, significant increase in body weights between
sample days (p > 0.05) and during recovery were observed (Table
3). However, 50 mg/kg BF produced a significant decrease in
organs weights, and at higher doses of 300 mg/kg ME and 150
mg/kg BF, there were significant decrease in organs weights and
significant increase in organ-brain weight ratio (P < 0.05) (Table
4). Therefore, the significant reduction in relative organs weights,
as well as significant increases in organ - brain weights ratio at
higher doses, suggest that at higher doses, which is often the
case with indiscriminate use of medicinal plants, there may be an
increased risk of organs toxicity [28]. Also, the general increase
in organ - brain weight ratio in recovery group compared to
toxicity group in all tested doses (Table 5) may suggest a form of
delayed toxicity [29].
Effect on hematological indices
Blood plays important roles in maintaining body functions
and homeostasis [30]. Generally, following single and repeated
dose administrations of either the extract or fraction (Table
6), the results showed varying degree of significant changes
(p < 0.05) in the hematological indices (WBC, RBC, Hb, HCT,
MCV, MCH, MCHC) and appear to be dose dependent and more
pronounced with BF (Table 6). In acute toxicity test, apart from
ME at 150 mg/kg, where RBC was significantly higher, other doses
showed significant decreases in RBC. Also, there were consistent
significant decreases in hemoglobin concentration, HCT, MCH and
MCHC in all tested doses of ME and BF. And while all the doses of
BF showed significant reduction in MCV, only at 150 mg/kg ME
was there such an effect observed. Meanwhile, upon repeated
administration of the test substances (ME and BF), there were
significant changes in RBC, MCV and MCH in all the tested doses
and only at 50 mg/kg BF was changes in MCHC not significant
(Table 6). However, while the pattern of changes were similar for
hemoglobin concentration, hematocrit and MCHC, the effect of
repeated doses on RBC, MCV, and MCH showed opposite pattern
between ME and BF, when comparing lower doses with higher
doses. For instance, RBC was significantly higher at lower dose of
100 mg/kg ME, and lower at higher dose of 300 mg/kg ME, but
the reverse was the case with BF, suggesting a different pattern
or mechanism of toxic action of ME and BF (Table 6). It should
be noted that significant changes in critical hematological indices
following acute and repeated doses, especially with BF and at
higher doses, is suggestive of potential toxicity [30,31]. Generally,
the differential dose dependent effects on various hematological
indices, may provide insights into the potential toxic effects
resulting from the cumulative doses of both the extract and
fraction. In addition, though the Recovery set showed significant
improvement in the hematological indices when compared with
Toxicity set, suggesting potential for recovery from toxic effects,
such recovery may take longer at higher doses, especially with
the butanol fraction (Table 7).
Citation: Shobo AA, Daniyan MO, Olayiwola G, et al. (2019) Toxicity Evaluation of the Extract and Fraction of Chrysophyllum
Albidum Seed Cotyledons in Rats. SOJ Pharm Sci, 6(1) 1-12. DOI: 10.15226/2374-6866/6/1/00194
Page 4 of 12
Copyright:
© 2019 Daniyan MO, et al.
Toxicity Evaluation of the Extract and Fraction of Chrysophyllum Albidum
Seed Cotyledons in Rats
Table 2: Changes in body weights following single and repeated dose oral administration of extract and fraction of C albidum seed cotyledon
Dose (mg/
Acute Toxicity**
kg)
Day 7 (%)
Dose /
Repeated Dose Toxicity#
Day 14 (%)
Groups
Day 7 (%)
Day 14 (%)
Day 21 (%)
Day 28 (%)
Control (n=5)
121.80 ± 1.36
131.20 ± 1.96
Control (n=5)
102.46 ± 0.54
104.76 ± 0.50
106.73 ± 1.08
109.27 ± 1.32
Methanol
150 (n=5)
103.00 ± 0.45*
116.00 ± 3.28*
100 (n=5)
103.21 ± 0.72
105.30 ± 0.65
108.03 ± 1.07
109.72 ± 0.99
Extract
300 (n=5)
107.20 ± 0.29*
110.40 ± 1.08*
300 (n=4)
103.03 ± 0.91
105.23 ± 0.73
107.01 ± 0.96
110.47 ± 1.28
600 (n=3)
104.40 ± 1.69*
118.40 ± 3.03*
Control (n=5)
121.80 ± 1.36
131.20 ± 1.96
Control (n=5)
102.46 ± 0.54
104.76 ± 0.50
106.73 ± 1.08
109.27 ± 1.32
40 (n=5)
106.00 ± 1.23*
112.80 ± 1.24*
50 (n=5)
102.74 ± 0.56
104.83 ± 0.78
106.44 ± 1.24
110.14 ± 1.49
80 (n=5)
105.20 ± 2.08*
109.40 ± 2.29*
150 (n=3)
103.36 ± 1.43
103.96 ± 1.01
105.22 ± 0.82
107.85 ± 2.12
160 (n=4)
94.00 ± 2.11*
94.80 ± 3.12*
Butanol
Fraction
*Significant difference at p < 0.05 when compared to Control. **Significant difference at p < 0.05 when comparing Day 7 to Day 14. #Significant
difference between Sampling Days using One Way ANOVA at p < 0.05.
Table 3: Changes in body weights of Recovery set of the repeated dose toxicity studies of extract and fraction of C albidum cotyledon
Dose / Groups
Methanol Extract
Last Read (Day 28)
Recovery (Post Day 28)#
Day 7 (%)
Day 14 (%)
Day 21 (%)
Control (n=5)
106.73 ± 1.08
116.07 ± 0.97
123.50 ± 0.86
131.35 ± 2.08
100 (n=5)
108.03 ± 1.07
94.45 ± 0.87*
101.25 ± 1.03*
109.05 ± 1.13*
300 (n=4)
107.01 ± 0.96
93.02 ± 0.96*
100.15 ± 0.96*
108.15 ± 0.96*
Control (n=5)
106.73 ± 1.08
116.07 ± 0.97
123.50 ± 0.86
131.35 ± 2.08
50 (n=4)
106.44 ± 1.24
87.36 ± 0.82*
93.44 ± 1.24*
102.54 ± 1.26*
150 (n=3)
105.22 ± 0.82
78.83 ± 0.78*
84.56 ± 1.01*
93.56 ± 1.01*
Butanol Fraction
*Significant difference at p <0.05 when compared to Control. #Significant difference at p < 0.05 Read. using One Way ANOVA followed by Dunnett’s
posthoc test comparing with Last
Table 4: Organ weights and Organ - brain weight ratio following single and repeated dose oral administration of extract and fraction of C. albidum
seed cotyledon
Dose
(mg/kg)
Organ weights (g)
Organ – brain weight ratio
Liver
Kidney#
Brain
Liver
Left kidney
Right kidney
Control (n=5)
4.02 ± 0.28
0.70 ± 0.02
0.91 ± 0.18
4.49 ± 0.44
0.39 ± 0.04
0.37 ± 0.03
150
3.57 ± 0.11
0.64 ± 0.03
0.88 ± 0.10
4.46 ± 0.88
0.39 ± 0.07
0.34 ± 0.02
3.63 ± 0.07
0.69 ± 0.02
0.94 ± 0.04
3.88 ± 0.22
0.35 ± 0.03
0.36 ± 0.01
3.45 ± 0.24
0.70 ± 0.02
0.98 ± 0.05
3.54 ± 0.20
0.37 ± 0.01
0.35 ± 0.02
Control (n=5)
4.02 ± 0.28
0.70 ± 0.02
0.91 ± 0.18
4.49 ± 0.44
0.39 ± 0.04
0.37 ± 0.03
40
3.81 ± 0.13
0.69 ± 0.05
0.95 ± 0.04
4.08 ± 0.29
0.36 ± 0.04
0.39 ± 0.04
3.83 ± 0.10
0.66 ± 0.02
0.92 ± 0.04
4.21 ± 0.26
0.37 ± 0.02
0.36 ± 0.02
4.22 ± 0.16
0.71 ± 0.05
0.94 ± 0.08
4.62 ± 0.47
0.37 ± 0.05
0.38 ± 0.05
(n=5)
Methanol
300
Extract
(n=5)
600
Acute
(n=3)
Toxicity
Butanol
Fraction
(n=5)
80
(n=5)
160
(n=4)
Citation: Shobo AA, Daniyan MO, Olayiwola G, et al. (2019) Toxicity Evaluation of the Extract and Fraction of Chrysophyllum
Albidum Seed Cotyledons in Rats. SOJ Pharm Sci, 6(1) 1-12. DOI: 10.15226/2374-6866/6/1/00194
Page 5 of 12
Copyright:
© 2019 Daniyan MO, et al.
Toxicity Evaluation of the Extract and Fraction of Chrysophyllum Albidum
Seed Cotyledons in Rats
Control (n=5)
4.00 ± 0.22
0.66 ± 0.02
1.04 ± 0.02
3.89 ± 0.26
0.32 ± 0.01
0.31 ± 0.01
Methanol
100
3.47 ± 0.08
0.66 ± 0.03
1.04 ± 0.05
3.39 ± 0.20
0.31 ± 0.03
0.33 ± 0.02
Extract
(n=5)
2.22 ± 0.26*
0.38 ± 0.05*
0.47 ± 0.03*
4.80 ± 0.10*
0.43 ± 0.03*
0.39 ± 0.03*
300
Repeated
(n=4)
Dose
Control (n=5)
4.00 ± 0.22
0.66 ± 0.02
1.04 ± 0.02
3.89 ± 0.26
0.32 ± 0.01
0.31 ± 0.01
Butanol
50
2.85 ± 0.21*
0.49 ± 0.02*
0.83 ± 0.02*
3.86 ± 0.28
0.31 ± 0.07
0.29 ± 0.07
Fraction
(n=5)
1.60 ± 0.08*
0.28 ± 0.07*
0.35 ± 0.02*
4.84 ± 0.13*
0.41 ± 0.03*
0.38 ± 0.02*
Toxicity
150
(n=3)
*Significant at p < 0.05 when compared to control group. Data are expressed as mean ± SEM. #indicate combined Relative weights of the two kidneys.
Table 5: Comparison of the Organs weights and Organ - Brain weight ratio of animals in repeated dose toxicity and recovery Sets
Organ Weights (g)
100 mg/kg (n=5)
Methanol
Extract
Organ - Brain Weight Ratio
300 mg/kg (n=4)
Toxicity
Recovery
Toxicity
Recovery
Liver
4.69 ± 0.11
6.90 ± 0.47*
3.28 ± 1.45
4.25 ± 1.75
Kidneys#
0.89 ± 0.03
1.08 ± 0.08
0.53 ± 0.22
0.62 ± 0.26
Brain
1.40 ± 0.07
0.99 ± 0.18
0.83 ± 0.34
0.73 ± 0.31
100 mg/kg (n=5)
300 mg/kg (n=4)
Toxicity
Recovery
Toxicity
Recovery
Liver
3.39 ± 0.20
4.68 ± 1.22
4.80 ± 0.10
5.82 ± 1.50
Left kidney
0.31 ± 0.03
0.35 ± 0.09
0.43 ± 0.03
0.46 ± 0.11
Right
0.33 ± 0.02
0.37 ± 0.10
0.39 ± 0.03
0.40 ± 0.11
kidney
50 mg/kg (n=4)
Butanol
Fraction
150 mg/kg (n=3)
Toxicity
Recovery
Toxicity
Recovery
50 mg/kg (n=4)
150 mg/kg (n=3)
Toxicity
Recovery
Toxicity
Recovery
Liver
4.02 ± 1.03
7.13 ± 0.43*
2.36 ± 1.45
4.47 ± 1.90
Liver
3.56 ± 0.28
5.05 ± 0.34*
4.84 ± 0.13
5.26 ± 1.28
Kidneys#
0.49 ± 0.12
0.78 ± 0.04
0.28 ± 0.17
0.69 ± 0.29
Left Kidney
0.31 ± 0.07
0.35 ± 0.03
0.41 ± 0.03
0.44 ± 0.10
Brain
1.14 ± 0.29
1.41 ± 0.06
0.51 ± 0.32
0.85 ± 0.35
Right
0.29 ± 0.07
0.30 ± 0.03
0.38 ± 0.02
0.42 ± 0.10
Kidney
*Significant at P < 0.05 when compared to Test group. Data are expressed as mean ± SEM
Table 6: Hematological parameters of animals administered single and repeated doses of extract and fraction of C. albidum seed cotyledon
Acute Toxicity
Hematological Parameters
Control
(n = 5)
Methanol Extract (mg/kg)
150 (n=5)
300 (n=5)
600 (n=3)
Butanol Fraction (mg/kg)
40 (n=5)
80 (n=5)
160 (n=4)
White Cell count (103 / µl)
6.3 ± 0.26
6.36 ± 0.25
5.88 ± 0.23
8.02 ± 0.26*
6.42 ± 0.23
8.1 ± 0.16*
4.78 ± 0.30*
Red Cell count (106 / µl)
4.52 ± 0.04
4.92 ± 0.06*
3.86 ± 0.05*
3.63 ± 0.06*
4.22 ± 0.04*
4.35± 0.02*
3.78 ± 0.06*
Hemoglobin (g/dL)
18.74 ± 1.44
13.22 ± 1.04*
13.7 ± 0.44*
13.48 ± 0.62*
13.46 ± 0.86*
13.68 ± 1.63*
12.33 ± 1.36*
Hemocrit (%)
50.2 ± 3.12
39.6 ± 3.15*
41 ± 1.34*
40.5 ± 1.87*
40.4 ± 2.56*
41 ± 2.45*
37 ± 2.03*
MCV (fl)
111.06 ± 3.2
80.49 ± 2.52*
106.22 ± 2.7
111.57 ± 3.8
95.73 ± 3.8*
94.25 ± 3.14*
97.88 ± 3.96*
MCH (pg)
41.46 ± 1.10
26.87 ± 1.05*
35.49 ± 1.13*
37.13 ± 1.56
31.90 ± 1.57*
31.45 ± 1.06*
32.62 ± 4.12*
MCHC (g/L)
37.33 ± 0.33
33.38 ± 0.22*
33.41 ± 0.12*
33.28 ± 0.21*
33.32 ± 0.23*
33.37 ± 0.41*
33.32 ± 0.13*
Citation: Shobo AA, Daniyan MO, Olayiwola G, et al. (2019) Toxicity Evaluation of the Extract and Fraction of Chrysophyllum
Albidum Seed Cotyledons in Rats. SOJ Pharm Sci, 6(1) 1-12. DOI: 10.15226/2374-6866/6/1/00194
Page 6 of 12
Copyright:
© 2019 Daniyan MO, et al.
Toxicity Evaluation of the Extract and Fraction of Chrysophyllum Albidum
Seed Cotyledons in Rats
Repeated Dose Toxicity
Hematological Parameters
Control
Methanol Extract (mg/kg)
Butanol Fraction (mg/kg)
(n = 5)
100 (n=5)
300 (n=4)
50 (n=5)
150 (n=3)
6.08 ± 0.31
5.42 ± 0.43
6.2 ± 0.20
6.43 ± 0.11
6.3 ± 0.11
Red Cell count (106 / µl)
3.02 ± 0.09
8.78 ± 0.60*#
1.77 ± 0.14*
2.55 ± 0.02*#
5.77 ± 0.23*
Hemoglobin (g/dL)
13.66 ± 1.49
16.12 ± 0.52
13.57 ± 1.30
14.9 ± 0.43#
13.47 ± 0.43
Hemocrit (%)
41.2 ± 2.42
42.2 ± 2.55#
30 ± 1.64*
45 ± 2.02#
34 ± 1.08*
MCV (fl)
136.42 ± 3.14
48.06 ± 1.76*#
169.49 ± 6.1*
176.47 ± 2.36*#
58.93 ± 2.92*
MCH (pg)
45.23 ± 2.16
18.36 ± 0.93*#
76.6 ± 4.3*
58.43 ± 2.00*#
23.34 ± 1.45*
MCHC (g/L)
33.16 ± 0.48
38.20 ± 1.08*#
45.23 ± 0.67*
33.10 ± 0.13#
39.62 ± 0.52*
White Cell count (103 / µl)
*Significant at P < 0.05 when compared to control. #Significant at P < 0.05, when comparing lower dose vs higher dose in the repeated dose Toxicity
study. Data are expressed as mean ± SEM
Table 7: Comparison of the hematological indices of animals in the repeated dose toxicity and recovery Sets (methanol extract and Butanol Fraction)
Hematological
Parameters
Methanol Extract (mg/kg)
100 (n=5)
Butanol Fraction (mg/kg)
300 (n=4)
50 (n=4)
150 (n=3)
Toxicity
Recovery
Toxicity
Recovery
Toxicity
Recovery
Toxicity
Recovery
White Cell count
(x 103 / µl)
5.42 ± 0.43*
3.62 ± 0.06
6.2 ± 0.20*
1.8 ± 0.05
6.43 ± 0.11*
2.12 ± 0.07
6.3 ± 0.11*
4.5 ± 0.10
Red Cell count (x
106 / µl)
8.78 ± 0.60
7.9 ± 0.84
1.77 ± 0.14*
3.1 ± 0.16
2.55 ± 0.02*
5.48 ± 0.04
5.77 ± 0.23*
4.83 ± 0.12
Hemoglobin (g/
dL)
16.12 ± 0.52
14.88 ± 0.42
13.57 ± 1.30
13.63 ± 1.13
14.9 ± 0.43*
13.46 ± 0.33
13.47 ± 0.43
14.37 ± 0.23
Hemocrit (%)
42.2 ± 2.55*
51.4 ± 2.15
30 ± 1.64*
56.67 ± 1.20
45 ± 2.02
50.4 ± 1.20
34 ± 1.08*
47.67 ± 2.16
MCV (fl)
50.44 ± 1.76*
70.06 ± 2.34
174.8 ± 6.1
192.5 ± 6.68
176.45 ± 2.36*
93.9 ± 1.49
65.2 ± 2.92*
113.03 ± 6.12
MCH (pg)
18.62 ± 0.93
20.2 ± 1.68
79.13 ± 4.3*
45.7 ± 2.35
58.5 ± 2.00*
25.16 ± 1.13
28.8 ± 1.45
33.63 ± 2.25
MCHC (g/L)
39.64 ± 1.08*
29.34 ± 2.23
45.17 ± 0.67*
24.17 ± 2.42
33.525 ± 0.13*
26.7 ± 0.39
40.83 ± 0.52*
31.63 ± 2.34
Effect on biochemical indices
We also examined the effects of the extract and fraction on
plasma levels of AST, ALT, creatinine and bilirubin. Elevated levels
of AST, ALT, and bilirubin have been reported to be indicators of
underlying cellular injuries [29,32,33]. The single and repeated
dose administrations of the extract or fraction showed varying
degree of significant changes (p < 0.05) in the biochemical indices
(Table 8). Interestingly, following both acute and repeated doses,
there were consistent dose dependent significant changes in ALT
and AST activities, and creatinine and direct bilirubin level, at all
tested doses of ME and BF when compared with control (Table
8). These changes were more pronounced with repeated doses,
suggesting more toxic responses to cumulative doses of the test
agents.
Generally, observed changes in assayed biochemical indices
indicate that BF could be more toxic and that there is a potential
for increased cumulative toxic effects at higher doses (Table 8).
While these changes are more pronounced with BF, the observed
significant increase in ALT activities and direct bilirubin, (Table 4)
may imply a potential to cause hepatocellular injury, possibly due
to increased workload on the liver [32,33], although, it is unclear
if this injury is sufficient to cause leakage in the mitochondrial
AST enzyme [29]. Also, the significant increase in creatinine
levels at all doses of ME and 80 mg/kg BF (Table 8), may be a
reflection of an impaired kidney function [29,32,33]. Additionally,
the sustained elevated level of DBIL at all tested doses of ME and
BF following period of recovery (Table 9), may be a reflection of
delayed toxicity and an indication of a potential for irreversibility
of toxic effects.
Citation: Shobo AA, Daniyan MO, Olayiwola G, et al. (2019) Toxicity Evaluation of the Extract and Fraction of Chrysophyllum
Albidum Seed Cotyledons in Rats. SOJ Pharm Sci, 6(1) 1-12. DOI: 10.15226/2374-6866/6/1/00194
Page 7 of 12
Copyright:
© 2019 Daniyan MO, et al.
Toxicity Evaluation of the Extract and Fraction of Chrysophyllum Albidum
Seed Cotyledons in Rats
Table 8: Changes in plasma biochemical indices following single and repeated doses administration of the extract and fraction of C. albidum seed
cotyledon
Acute Toxicity
Parameters
Control
Methanol Extract (mg/kg)
Butanol Fraction (mg/kg)
(n = 5)
150 (n=5)
300 (n=5)
600 (n=3)
40 (n=5)
80 (n=5)
160 (n=4)
ALT (IU/L)
24.40 ± 2.16
29.60 ± 2.02
32.80 ± 1.86*
33.20 ± 2.09*
29.80 ± 2.42
31.80 ± 1.52*
69.40 ± 3.21*
AST (IU/L)
58.40 ± 2.90
51.00 ± 3.32
48.20 ± 2.50*
40.20 ± 2.20*
53.00 ± 2.45
46.00 ± 1.73*
38.20 ± 1.56*
Creatinine (g/dl)
1.36 ± 0.09
2.02 ± 0.13*
3.54 ± 0.15*
2.82 ± 0.12*
1.27 ± 0.06
1.68 ± 0.07*
0.95 ± 0.03*
Direct Bilirubin (g/dl)
0.03 ± 0.01
0.10 ± 0.06
0.41 ± 0.07*
0.09 ± 0.01*
0.04 ± 0.01
0.16 ± 0.04*
0.30 ± 0.02*
Indirect Bilirubin (g/dl)
0.66 ± 0.04
0.55 ± 0.08
0.28 ± 0.03*
0.53 ± 0.08
0.70 ± 0.06
0.60 ± 0.06
0.24 ± 0.01*
Total Bilirubin (g/dl)
0.69 ± 0.03
0.59 ± 0.09
0.51 ± 0.07*
0.62 ± 0.07
0.74 ± 0.05
0.75 ± 0.03
0.53 ± 0.04*
Repeated Dose Toxicity
Parameters
Control (n = 5)
Methanol Extract (mg/kg)
Butanol Fraction (mg/kg)
100 (n=5)
300 (n=4)
50 (n=5)
150 (n=3)
ALT (IU/L)
40.20 ± 0.49
49.00 ± 2.83*#
26.60 ± 1.88*
28.80 ± 1.26*#
19.40 ± 1.88*
AST (IU/L)
34.60 ± 2.98
60.80 ± 2.43*#
22.00 ± 2.33*
26.40 ± 2.24*#
13.40 ± 1.24*
Creatinine (g/dl)
0.75 ± 0.08
1.18 ± 0.06*
1.05 ± 0.04*
0.63 ± 0.02*#
0.31 ± 0.02*
Direct Bilirubin (g/dl)
0.05 ± 0.01
0.06 ± 0.01
0.03 ± 0.01
0.08 ± 0.02
0.03 ± 0.01
Indirect Bilirubin(g/dl)
0.55 ± 0.05
0.44 ± 0.03#
0.17 ± 0.05*
0.39 ± 0.03*#
0.24 ± 0.03*
Total Bilirubin (g/dl)
0.60 ± 0.04
0.50 ± 0.02#
0.20 ± 0.03*
0.47 ± 0.02*#
0.26 ± 0.06*
*P < 0.05; Dose (s) vs. Control group; #P < 0.05, lower dose vs. higher dose. Data are expressed as mean ± SEM
Table 9: Comparison of the plasma biomarkers of animals in the repeated dose toxicity and recovery Sets (methanol extract and butanol fractions)
Biomakers
Methanol Extract (mg/kg)
100 (n=5)
Butanol Fraction (mg/kg)
300 (n=4)
50 (n=4)
150 (n=3)
Toxicity
Recovery
Toxicity
Recovery
Toxicity
Recovery
Toxicity
Recovery
ALT (IU/L)
49.00 ± 2.83
42.00 ± 1.52
26.60 ± 1.88
29.00 ± 1.85
28.80 ± 1.26*
42.60 ± 1.25
19.40 ± 1.88*
27.80 ± 1.37
AST (IU/L)
60.80 ± 2.43*
45.80 ± 2.00
22.00 ± 2.33
18.20 ± 1.77
36.40 ± 2.24
40.00 ± 2.80
13.40 ± 1.24*
21.00 ± 1.87
Creatinine (g/dl)
1.18 ± 0.06
1.30 ± 0.06
1.05 ± 0.04*
0.81 ± 0.04
0.79 ± 0.02
0.82 ± 0.04
0.31 ± 0.02*
0.57 ± 0.04
Direct Bilirubin
0.16 ± 0.01
0.15 ± 0.02
0.23 ± 0.01
0.20 ± 0.02
0.18 ± 0.02
0.19 ± 0.02
0.43 ± 0.01
0.46 ± 0.03
0.44 ± 0.03*
0.31 ± 0.01
0.17 ± 0.05*
0.45 ± 0.02
0.39 ± 0.03*
0.51 ± 0.04
0.24 ± 0.03
0.19 ± 0.01
0.60 ± 0.02*
0.46 ± 0.02
0.40 ± 0.03*
0.65 ± 0.02
0.57 ± 0.02*
0.69 ± 0.05
0.67 ± 0.02
0.65 ± 0.02
(g/dl)
Indirect
Bilirubin (g/dl)
Total Bilirubin
(g/dl)
*P < 0.05; Toxicity group vs Recovery group. Data were expressed as mean ± SEM
Citation: Shobo AA, Daniyan MO, Olayiwola G, et al. (2019) Toxicity Evaluation of the Extract and Fraction of Chrysophyllum
Albidum Seed Cotyledons in Rats. SOJ Pharm Sci, 6(1) 1-12. DOI: 10.15226/2374-6866/6/1/00194
Page 8 of 12
Toxicity Evaluation of the Extract and Fraction of Chrysophyllum Albidum
Seed Cotyledons in Rats
Effect on organ histology
The inclusion of pathological findings as components of
pathology data in assessing oral toxicity is necessary to provide
a more holistic toxicological information [22,24]. Histological
findings following acute and repeated doses of the ME revealed
presence of pathological lesions in the FWRs kidneys and liver,
especially at higher doses (Figure 1 and Figure 2). Following acute
administration of 150, 300, 600 mg/kg ME, the kidney showed
presence of renal lesions, including hydropic degeneration
(Figure. 1). The liver showed an enhanced depiction of the
centrilobular vein, hepatocytes and sinusoids at 150 mg/kg
ME, and slight presence of hydropic changes at 300 mg/kg ME.
However, at 600 mg/kg ME, the presence of pathological lesions
including hydropic degeneration, microvesicular steatosis,
marked by small or large vacuoles can be observed (Figure. 1).
On the other hand, following 28-day repeated administration of
Copyright:
© 2019 Daniyan MO, et al.
ME and 21-day recovery period, histology of the kidney revealed
the presence of hydropic degeneration and mild vacuolar
degeneration at 100 mg/kg and 300 mg/kg ME respectively
in both Toxicity and Recovery sets. Also, while mild traces of
hepatocellular lesions was seen only in Toxicity set at 100 mg/
kg ME, at 300 mg/kg ME, both Toxicity and Recovery revealed a
cross section of hepatic parenchyma with hepatocellular lesions
(Figure. 2). Furthermore, except at 160 mg/kg, where presence
of pathological lesions including hydropic degeneration and
fatty changes were observed (Figure. 3), no other pathological
lesions was observed in kidney and liver following acute doses
of 40, 80 and 160 mg/kg BF. In addition, following repeated
administration of BF, the Toxicity set showed pathological lesion
such as hydropic degeneration at 50 mg/kg BF, and presence of
tubular and glomerular structure degeneration at 150 mg/kg BF
in kidney architecture (Figure. 4).
Figure 1: Effects of Histology of acute doses of methanol extract of C. albidum seed cotyledon on rat Kidney (K) and Liver (L). DCT, distal convoluted tubule; PCT, proximal convoluted tubule; G, glomerulus; H, hepatocyte; V, central vein. Red arrows were used to identify pathological changes
and vascular degeneration. Staining was done using H&E and magnification was x400.
Figure 2: Effects of 28-day repeated administration of methanol extract of C. albidum seed cotyledons (Toxicity - T), followed by 21 days
recovery period (Recovery - R) on rat Kidney (K) and Liver (L). DCT, distal convoluted tubule; PCT, proximal convoluted tubule; G, glomerulus;
US, urinary space; H, hepatocyte; V, central vein; A, hepatic artery; D, bile duct. Red arrows were used to identify pathological changes and vascular
degeneration All staining were done with H&E and Magnification was x400.
Citation: Shobo AA, Daniyan MO, Olayiwola G, et al. (2019) Toxicity Evaluation of the Extract and Fraction of Chrysophyllum
Albidum Seed Cotyledons in Rats. SOJ Pharm Sci, 6(1) 1-12. DOI: 10.15226/2374-6866/6/1/00194
Page 9 of 12
Toxicity Evaluation of the Extract and Fraction of Chrysophyllum Albidum
Seed Cotyledons in Rats
Copyright:
© 2019 Daniyan MO, et al.
Figure 3: Photomicrographs of rat Kidney (K) and Liver (L) following administration of the acute doses of butanol fraction of C. albidum
seed cotyledons. DCT, distal convoluted tubule; PCT, proximal convoluted tubule; G, glomerulus; H, hepatocyte; V, central vein; D, bile duct. Red arrows were used to identify pathological changes and vascular degeneration. All were stained using H&E and Magnification was x400.
Figure 4: Photomicrograph of rat Kidney (K) and Liver (L) following 28-day repeated administration of butanol fraction of C. albidum seed
cotyledons (toxicity - T), followed by 21 days recovery period (recovery - R). DCT, distal convoluted tubule; PCT, proximal convoluted tubule; G,
glomerulus; US, urinary space; H, hepatocyte; V, central vein; A, hepatic artery; D, bile duct. Red arrows were used to identify pathological changes and
vascular degeneration All staining were done with H&E and Magnification was x400.
In general, the presence of pathological lesions (hydropic
degeneration and microvesicular steatosis) in the histology of the
FWR kidney and liver following acute and repeated doses of the
ME and BF may be representative of the potential negative effect
of C. albidum seed cotyledons on these organs. The observed
histologic changes could be morphologic correlates of reversible
cell injury [34]. Meanwhile, though observed pathological
changes were maintained in the Recovery set of ME at 300 mg/
kg, suggesting persistent form of toxicity, the gradual reduction
and/or complete eradication of pathological changes following
recovery at other tested doses of both ME and BF are indicative
of a pattern of reversible form of toxicity, and may be a reflection
of the non-persistence of the negative effect on the organs [34].
Conclusion
The many useful properties of Chrysophyllum albidum
[14–17] is a pointer to its potential as a natural source of drug,
but also makes it a subject of potential abuse, especially among
the local consumers. Toxicological findings from this study
revealed that the test materials (ME and BF of C. albidum seed
cotyledons) showed potential to induced toxicity with some level
of persistent signs of intoxication following acute and repeated
dose administration. This is consistent with their relatively low
LD50 values, observed increase mortality with increasing doses
and significant, though relatively reversible, changes in most
hematological, biochemical and histological data. Hence, while
taking advantage of its many economic and medicinal properties,
there is the need to protect against indiscriminate uses and to
apply caution in the use of the seed cotyledon of C. albidum.
Citation: Shobo AA, Daniyan MO, Olayiwola G, et al. (2019) Toxicity Evaluation of the Extract and Fraction of Chrysophyllum
Albidum Seed Cotyledons in Rats. SOJ Pharm Sci, 6(1) 1-12. DOI: 10.15226/2374-6866/6/1/00194
Page 10 of 12
Toxicity Evaluation of the Extract and Fraction of Chrysophyllum Albidum
Seed Cotyledons in Rats
Acknowledgments
We acknowledge the assistance of the following on
histopathology aspects of the work and its analysis: Dr. O. Oladele
of the Department of Morbid Anatomy and Forensic Medicine,
Obafemi Awolowo University Teaching Hospitals Complex
(OAUTHC), Ile – Ife, Nigeria, Dr. A. Onaolapo and Mrs. M.O Cyril of
Department of Anatomy, Ladoke Akintola University, Ogbomoso,
Nigeria.
Funding
This research did not receive any specific grant from funding
agencies in the public, commercial, or not-for-profit sectors.
Declarations
Ethical Approval: The protocol was approved by the Committee
on the care and use of laboratory animals, Obafemi Awolowo
University, Ile-Ife, Nigeria (Protocol number PHP12/13/H/0601).
Clinical trial registration: N/A
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Albidum Seed Cotyledons in Rats. SOJ Pharm Sci, 6(1) 1-12. DOI: 10.15226/2374-6866/6/1/00194
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Citation: Shobo AA, Daniyan MO, Olayiwola G, et al. (2019) Toxicity Evaluation of the Extract and Fraction of Chrysophyllum
Albidum Seed Cotyledons in Rats. SOJ Pharm Sci, 6(1) 1-12. DOI: 10.15226/2374-6866/6/1/00194
Page 12 of 12
Abiodun Ogundaini