Content uploaded by Emelia Kisseih
Author content
All content in this area was uploaded by Emelia Kisseih on Sep 19, 2021
Content may be subject to copyright.
Bekoe et al., IJPSR, 2021; Vol. 12(1): 515-523. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 515
IJPSR (2021), Volume 12, Issue 1 (Research Article)
Received on 26 October 2019; received in revised form, 22 June 2020; accepted, 11 July 2020; published 01 January 2021
EVALUATING PHARMACOGNOSTIC PARAMETERS AND MUTAGENIC ACTIVITY OF
KHAYA SENEGALENSIS (DESR.) A. JUSS.
E. Oppong Bekoe * 1, C. Kitcher 1, J. A. Sarkodie 1, K. B. Dodoo 1, S. Frimpong-Manso 3 and M. C.
Toafode 3
Department of Pharmacognosy and Herbal Medicine 1, Department of Pharmaceutical Chemistry 2, School
of Pharmacy, University of Ghana, Ghana.
University of IRGIB Africa 3, Inter-Regional University of Industrial Engineering, Biotechnologies and
Applied Sciences, Cotonou, Benin.
ABSTRACT: The stem bark of Khaya senegalensis, known as mahogany is
commonly sold on the Ghanaian market for the treatment of numerous ailments.
To ensure its safe and efficacious use, there is a need to ensure that this plant
material is of the best quality. Quality will involve ensuring its identity, purity
and content are up to recommended standards by applying available and relevant
methods. As a means to improve the safe use of the material, mutagenic studies
were also conducted. This study sought to evaluate parameters that can be easily
applied to evaluate the quality of the stem bark of K. senegalensis. Macroscopy,
microscopy, physicochemical, and phytochemical investigations using fluore-
scence and UV analysis were employed. Microscopic analysis of K. senegalensis
leaf revealed the presence of calcium oxalate crystals, trichomes, stomata cells,
and vein islets. Phytochemical screening of the stem bark showed the presence
of alkaloids, saponins, tannins, flavonoids, glycosides, and terpenes. UV analysis
displayed spectra that shift and change in absorbance on acidification and
basification of the aqueous and ethanol extracts. Under UV light of short and
long wavelengths, changes in fluorescing of the extracts were observed in
various solvents. The characteristic macroscopic, microscopic, physicochemical,
and chemical parameters evaluated for K. senegalensis can be employed in the
simple authentication and establishment of the quality of raw materials of this
plant. This will improve the quality and hence efficacy of K. senegalensis when
used for treating diseases and manufacturing medicines.
INTRODUCTION: Herbal remedies are used
worldwide for both minor and serious ailments 1-3.
Due to poverty and limited access to contemporary
medicine, about 80% of the world’s population is
believed to use herbal remedies as their source of
primary healthcare 4-6.
QUICK RESPONSE CODE
DOI:
10.13040/IJPSR.0975-8232.12(1).515-23
This article can be accessed online on
www.ijpsr.com
DOI link: http://dx.doi.org/10.13040/IJPSR.0975-8232.12(1).515-23
Herbal medicines are in great demand, especially in
developing countries such as Ghana, for primary
healthcare. This is because of their efficacy, safety,
low cost, and lesser side effects. However, the non-
availability of pharmacopoeial standards with its
requisite analytical equipment for plant-derived
medicines is a major drawback that frustrates
clinical trials and thus overshadows the time tested
healing properties of plant medicines 7.
In this respect, K. senegalensis, which is one of the
medicinal plant sold in high quantities on the
Ghanaian market 8, 9 and is also a component of
many Pharmacopoeial medicinal preparations of
Keywords:
Khaya senegalensis, Raw material,
Quality control, Simple methods
Correspondence to Author:
Dr. Emelia Oppong Bekoe
Senior Lecturer,
Department of Pharmacognosy
and Herbal Medicine, School of
Pharmacy, University of Ghana, P.O.
Box LG 43, Legon, Accra, Ghana.
E-mail: emekisseih@yahoo.co.uk
Bekoe et al., IJPSR, 2021; Vol. 12(1): 515-523. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 516
multiple African groups 10, was evaluated for its
quality control parameters. This will ensure the
quality of the raw plant materials in a setting such
as Ghana.
Khaya senegalensis (Desr.) A. Juss is of the family
Meliaceae. It is commonly known as African
mahogany. It is an evergreen tree that grows up to a
height of 18 m high, up to 1 m in diameter, with a
clean bole, 6-9 m long. Buttresses are not
conspicuous or absent. It is cultivated as avenue
trees in many towns and cities 11. This plant is
commonly formulated as decoctions and tinctures. K.
senegalensis is used extensively in West Africa as a
bitter tonic, febrifuge, blood tonic and antimalarial
8. The extract from the bark has analgesic,
anticonvulsant, anthelmintic (human and
veterinary), antimicrobial, antipyretic, hematinin;
febrifuge, sedative and emetic properties. It is used
in the treatment of jaundice, scorpion bites,
allergies, dermatoses, infections of the gums,
wounds, constipation, chronic weakness, headache,
heat rash, loss of appetite, malaria, ulcer, and
sexually transmitted disease 11. It is known to be a
potential arbortifacient 12, 13.
The antipyretic, anticonvulsive, analgesic, and
sedative properties are believed to be due to the
simple coumarins (e.g., scopoletins) present in the
stem bark. Aqueous extracts of the stem bark and
leaves containing limonoid have also demonstrated
a stronger antisickling activity 13, 14. In addition to
the bark, both the seeds and leaves have also been
used as medicines for treating fever and headache,
whilst preparations made from the roots have been
used against syphilis, leprosy and as an aphrodisiac
15.
On the other hand, this plant has also been shown
to exhibit some toxicity. A study indicates that the
aqueous stem bark extract of K. senegalensis may
affect the cellular integrity of vital organs of the
body 11, 16. Renal toxicological assessments of
sulphonated nanocellulose isolated from Khaya
sengalensis seed has also been shown to potentially
toxic in rats 17.
Various parts of this plant have various other non-
medicinal uses such as fodder, fuel, fiber, timber,
gum and resin dyestuff, oils and even as fish poison
11. The bark can be used for tanning leather as it is
known to contain high quantities of tannins 18, 19.
Also, the current widespread increase in the use of
herbal medicines have arisen with it immense
safety concerns. For example, the consistent
increase in cancer cases in Ghana has led to the
advancement of various theories with regards to the
causes. Among the possible proposed causes are
commonly used substances, which may include
some herbal medicines. This notion may not be
totally wrong since some medicinal plant products
have been proven to contain components that are
potential mutagens 20-24. It is also important to note
that most of such traditionally used medicinal
plants have not been subjected to exhaustive
toxicological investigations. Only a small
proportion of such plants have been thoroughly
investigated scientifically for their acclaimed
benefits and side effects 25. Reports have it that a
number of such plants used as food or in traditional
medicines may exhibit in-vitro mutagenicity 26-28
and 90% of such mutagens could be carcinogens
capable of inducing cells to undergo abnormal
growth and genetic defects 28, 29, hence the rational
to perform the mutagenic test on such a widely
used plant. To screen for mutagenicity, the Ames
test which is very commonly employed because it
enables the screening of many chemicals rapidly
and inexpensively was used 30, 31. If found to be
mutagenic, this can be further investigated in
animal model 32.
MATERIALS AND METHODS:
Plant Material: The leaves and stem barks of K.
senegalensis were collected from the botanical
garden of the University of Ghana. The samples
were authenticated at the Ghana Herbarium,
Department of Plant and Environmental Biology,
University of Ghana. A voucher specimen (number
PSM68/19) have been kept in the herbarium of the
Department of Pharmacognosy and Herbal Medicine,
University of Ghana, Legon. Fresh leaves were
obtained, washed with water, and stored in glycerin
to be later used for microscopic analysis. The stem
bark was air-dried for three weeks and milled into
coarse powder, and kept in well-labeled air-tight
containers for further analysis.
Macroscopic Evaluation: The dried stem bark of
K. senegalensis was preliminarily analyzed for
color, odor, texture, and fracture, while leaves were
analyzed for their shape, color, arrangement, apex
base, texture, margin, and venation.
Bekoe et al., IJPSR, 2021; Vol. 12(1): 515-523. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 517
Microscopic Evaluation: The microscopic
characteristics of the leaves were studied with a
Leciad compound light microscope using procedures
recommended by WHO guidelines on Quality
control methods for herbal materials, 2011 33. Four-
millimeter square (4 mm2) sizes of the mature
lamina of the leaves were cut with a microtome.
The lamina was then boiled for about two hours in
chloral hydrate solution to clear the chlorophyll
content. The sections were mounted on a clean
glass slide with the help of glycerin and covered
with a coverslip. The slides were observed under a
light microscope at an objective magnification of
x10. The presence of calcium oxalate crystals,
stomata, and trichomes was observed. Quantitative
parameters such as vein islet number, veinlet
termination number, stomata number, and stomatal
index were calculated according to standard
protocols as described by WHO, 2011.
Powdered Microscopy: Small quantities of milled
stem bark powder were mounted in 2-3 drops of
chloral hydrate on a glass slide. Each slide was
covered with a coverslip and examined under the
microscope. The samples were photographed for
lignified fibers, calcium oxalate crystals, and
stomatal cells. For the observation of lignified
tissues, a small quantity of the stem bark was
mounted in phloroglucinol and concentrated HCl
and observed under low power. Brachysclereids,
which were stained pink to purple, were identified
and also photographed.
Physico-chemical Analysis: Total ash, acid
insoluble, and water-soluble ash values were
estimated, and extractive values of petroleum ether,
50 % ethanol, and water were also determined for
the air-dried stem bark. Furthermore, the foreign
organic matter, swelling index, and the foaming
index were analyzed according to methods
described by WHO, 2011 33.
Preliminary Phytochemical Screening: Preliminary
phytochemical screening was performed to detect
the presence of alkaloids, saponins, tannins,
flavonoids, glycosides, and terpenes as per the
methods of Khandelwal, 2002 and Harbourne, 1992
34, 35.
Fluorescence Studies: Characteristic fluorescence
of the powdered stem bark of K. senegalensis was
carried out in distilled water, dilute NaOH, dilute
HCl, methanol, ethanol as well as the respective
acidified and basified solvents. This was performed
according to methods described in published
literature 36. Observations of the samples were
made under visible daylight and UV light of short
wavelength (λ254 nm) and UV light of long-
wavelength (365 nm) for their characteristic colors
37.
Ultraviolet/Visible Spectrometric Analysis: An
aqueous extract was prepared by weighing 10 g of
coarsely powdered air-dried stem bark of K.
senegalensis into 100 mL of distilled water. The
plant material were sonicated three times for
intervals of 15 min each. The solution was filtered
into a round-bottomed flask and concentrated using
a rotary evaporator set at 45 ºC. The residue was
freeze-dried to obtain the crude aqueous extract. An
amount of 20 mg of this extract was accurately
weighed and separately dissolved in 20 mL of
distilled water, dilute hydrochloric acid (0.1M),
dilute sodium hydroxide (0.1M), methanol,
acidified methanol, and basified methanol. Each of
these preparations was sonicated for 10 min to
ensure complete dissolution of the extracts. Each of
the sonicated solutions was allowed to cool to room
temperature and filtered using membrane filter
0.45μm. Dilutions of 1 in 10 of each of the solution
were made in the respective solvents. The diluted
solutions were filtered and analyzed using a 1 cm
quartz cell/cuvette (sample holder). All the samples
were scanned over a wavelength range of 200 –
600 nm (Ultraviolet/ visible range) using a single
beam Jenway 7315 UV- Vis spectrophotometer.
Heavy Metal Analysis: An Olympus Vanta
Portable ED-XRF (VMR) analyzer equipped with
4-Watt X-ray tube with application-optimized
anode material rhodium (Rh), tungsten (W), 50kV
x-ray tube, and large area silicon drift detector was
used to analyze the sample for heavy metals.
Calibration of the XRF was done using the SARM
2711A, certified reference material from the
manufacturer. Twelve (12 g) grams of the stem
bark powder was sieved with a sieve of 180 μm
mesh size into a fine powder and kept in a dry well-
labeled container for analysis. The loose sample
was irradiated following the manufacturer’s
protocol, and simultaneous measurement of the
levels of heavy metals present was performed.
Bekoe et al., IJPSR, 2021; Vol. 12(1): 515-523. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 518
Microbiological Analysis:
Bacterial and Fungal Counts: One gram (1 g) of
the powdered stem bark of K. sengalensis was
weighed into 100 mL of sterile water in a tube to
make a stock concentration. The tubes for the
dilution were labeled as: a (102), b (104), c (106), d
(108), e (1010). Using a micropipette, 1 mL of the
content of tube (a) was transferred to the tube (b)
containing 99 mL of sterile water and mixed
gently. The process was repeated until all the tubes
containing 99 mL of sterile water were mixed. An
aliquot of 1 mL of each content was taken and
added to their corresponding tubes of molten
nutrient agar at 45 ºC. Each tube was poured into
its labeled corresponding sterile petri dish (A, B, C,
D, and E). The petri dishes were inverted and
incubated at 37 ºC for 48 h and the number of
colonies counted. A similar procedure was
employed in estimating the fungal count in the
sample, but instead, potato dextrose agar was used.
The petri dishes were incubated upright at room
temperature for 5 days. Bacillus subtilis ATCC
6538-P and Candida albicans ATCC 2091 were
used as growth controls for the nutrient agar and
potato dextrose agar, respectively.
Mutagenic Studies:
Sample Preparation: Dried K. senegalensis stem
bark was pulverized and extracted by decoction.
Fifty grams (50.0 g) of pulverized material was
boiled in 500 mL of distilled water for 20 min. The
decoction was centrifuged at 6000 RPM for 10
min. The supernatant was collected and
concentrated under vacuum at 40 ºC and
lyophilized to obtain dried crude extracts. The
extract was stored at -20 ºC for use. The samples
were prepared and diluted with sterile water on the
day of the assay to a concentration of 10 µg/mL
and sterile filtered using a 0.22 µM membrane
filter.
Induction of Mutation: The Muta-ChromoPlateTM
kit (manufactured by Environmental Bio-Detection
Products Inc, Ontario, Canada), which works on the
principle of the Ames test 38 was employed to
determine the mutagenicity potential of the
extracts. The experiment was carried out in
accordance with protocol provided by the
manufacturer using Salmonella typhimurium TA98.
Similar methods are reported in published literature
36.
The level of mutagenicity was detected as an
increase in the number of histidine revertants with
reference to scores provided in the kit. The
statistical table provided in the kit was used to
compare the natural background rate of reverse
mutation to the rate of reverse mutation within a
sample assay. The number of positive wells scored
in the 96 well plates determined the significance of
mutation in the fluctuation test 39.
The statistical table provided in the kit was used to
compare the natural background rate of reverse
mutation to the rate of reverse mutation within a
sample assay. Based on these samples, p values of
0.001 were classified as highly mutagenic, p values
of 0.01 moderately mutagenic, and p values of 0.05
weakly mutagenic. Mutation in the TA 98 is
indicative of a frameshift mutation (FSM).
RESULTS AND DISCUSSION:
Macroscopic and Microscopic Characteristics:
A summary of the organoleptic and macroscopic
characteristics of both the leaves Table 1 and the
stem bark of K. senegalensis Table 2 are provided
below. Fig. 1 also displays the pinnately compound
leaves and unique red inner colored bark and grey
outer-bark of this plant. The estimated constants of
the leaves, such as vein islet number, veinlet
termination number, and paracytic stomata are
characteristics of the leaves which can be used in
the rapid authentication of the plant. This can be
used in identifying K. senegalensis before the bark
samples are harvested for processing. The
characteristic features of green-grey outer bark and
reddish-brown inner bark, characteristic odor, bitter
taste, and short-splintery fracture, which are simple
means of further confirming the identity and
quality of the stem bark of K. senegalensis.
TABLE 1: MACROSCOPIC CHARACTERISTICS OF
K. SENEGALENSIS LEAVES
Morphology
Description
Type
Shape
Pinnately compound
Oblong to obovate
Colour
Green
Arrangement
Alternate
Venation
Pinnately reticulate
Base
Apex
Decurrent
Mucronate
Margin
Entire
Surface
Glabrous
Texture
Smooth and papery
Bekoe et al., IJPSR, 2021; Vol. 12(1): 515-523. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 519
TABLE 2: ORGANOLEPTIC CHARACTERISTICS OF
THE STEM BARK OF K. SENEGALENSIS
Morphology
Description
Condition
Dried
Outer bark color
Dark-grey
Inner bark color
Reddish-brown
Odor
Slight
Taste
Astringent
Texture
Scaly
Fracture
Short-splintery
FIG. 1: LEAVES AND STEM BARK OF K. SENEGALENSIS
Microscopic analysis of the leaves of K.
senegalensis showed an average vein islet number
of 24.9, vein islet termination number of 26,
stomata number of 5.3, and number of epidermal
cells of 173.3 per mm2 as indicated in Table 3. Fig.
2 of the leaves of K. senegalensis displays the
epidermal cells and cystoliths, veinlet terminations
and vein islets, calcium oxalate crystals, stomata
cells, palisade cells, and branched dendritic
trichomes.
TABLE 3: LEAF CONSTANTS
Parameter
K. senegalensis
Stomatal Number
5.3
Epidermal Cell Number
173.3
Vein islet Number
24.9
Veinlet termination number
26.0
The stomatal index [stomata per
square mm of epidermis]
2.97
FIG. 2: MICROSCOPIC CHARACTERISTICS OF LEAF EPIDERMIS AND POWDERED LEAF OF K. SENEGALENSIS. A.
HIGHLY BRANCHED TRICHOMES; B. WAVY EPIDERMAL CELLS AND CYSTOLITHS. C. VEINLET TERMINATIONS
AND VEIN ISLETS. D. PRISMATIC CALCIUM OXALATE CRYSTAL
FIG. 3: MICROSCOPIC FEATURES OF THE POWDERED STEMBARK OF K. SENEGALENSIS. A. SHOWS PRISMATIC
CALCIUM OXALATE CRYSTALS; B. SHOWS A CLUSTER OF BRACHYSCLEREIDS
Bekoe et al., IJPSR, 2021; Vol. 12(1): 515-523. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 520
Powdered Microscopy of Stem Bark: Microscopic
investigation of the powdered stem bark revealed
the presence of prismatic calcium oxalate crystals,
which were in abundance, as well as brachy-
sclereids, which were occurring in clusters (refer to
Fig. 3).
Physico-chemical and Phytochemical analysis:
Physico-chemical constants that were determined
for the bark of K. senegalensis included the ash
values and foaming index of 142.8. Details are
provided in Table 4. The air-dried stem bark had
an average moisture content of 5.47% w/w,
indicating that a significantly higher moisture
content than this value could be indicative of a poor
quality material because this could hasten moisture
and microbial decomposition which detrimentally
affect the quality. K. senegalensis had no swelling
index, possibly indicating the absence of mucilage,
pectin or hemicellulose which have swelling
properties 40. Its foaming index was 142, thus
indicating the possible presence of saponins which
caused persistent foam when the aqueous decoction
is shaken 40.
TABLE 4: ASH VALUE PARAMETERS
Parameters
K. senegalensis (% w/w)
Total Ash
6.53
Acid insoluble ash
2.13
Water-soluble ash
6.6
Moisture content
5.47
Foreign organic matter
Nil
Foaming index
142.85
Swelling index
-
K. senegalensis yielded extractive values of 11.8,
15.1, and 1.9% w/w for water, ethanol, and
petroleum ether, respectively. K. senegalensis had a
higher proportion of middle polar constituents
(11.8% w/w in ethanol) significantly as compared
to polar constituents in water (11% w/w) and non-
polar constituents in petroleum ether (15% w/w).
The extracts tested positive for alkaloids, tannins,
glycosides, saponins, flavonoids, and steroids.
TABLE 5: EXTRACTIVE VALUES
Parameters
K. senegalensis (% w/w)
Petroleum ether
1.98
50% ethanol
15.16
Water
11.80
Fluorescence Analysis: Analysis for characteristic
fluorescence of the plant material at the short, and
long ultra-violet wavelengths of λ 254 nm and λ
365 nm showed colors as provided in Table 6.
Fluorescence is the ability of a substance to emit
light that has been absorbed. The light emitted
depends on the wavelength of emission and the
type of compound involved. This can, therefore, be
used to qualitatively assess the components of
herbal medicines. For crude herbal drugs, this can
be seen in the form of bright colors under short or
long wavelengths of ultra-violet light. The
characteristic fluorescence of K. senegalensis at
these wavelengths could be used as a further
confirmation of the identity, purity, and quality of
the plant sample. For example, in methanol the
plant sample is wine under daylight, deep blue
under at 254, and ash at 365 nm.
TABLE 6: FLUORESCENT STUDIES OF POWDERED
STEM BARK OF K. SENEGALENSIS IN VARIOUS
SOLVENTS
Daylight
254 nm
365 nm
Powdered Sample
only
Reddish-
brown
Dark brown
Dark brown
Distilled water
Light brown
Deep blue
Pale blue
1N HCl
Orange
Deep blue
Pale blue
1N NaOH
Black
Deep blue
Pale blue
50 % H2SO4
Dark Brown
Deep blue
Pale blue
Methanol
Wine
Deep blue
Ash
Glacial acetic acid
Orange
Purple
Light purple
Nitric acid
Wine
Deep blue
Light purple
Chloroform
Dark brown
Deep blue
Light purple
50 % FeCl3
Deep green
Deep blue
Pale blue
95 % Ethanol
Cherry red
Deep blue
Pale blue
UV Spectra Analysis: The UV spectra of the
aqueous extract of the stem bark of K. senegalensis
dissolved in various solvents of different pHs,
showed spectra shifts under acidified and basic
conditions and changes in absorbance levels (refer
to Fig. 4). This indicates that component (s) of the
crude extract of K. senegalensis absorb ultraviolet/
visible light of a wavelength of 233 and 280 nm.
There are slight shifts in λmax for the basified
extract in dilute NaOH and in HCl but none in
Methanol. Similar patterns are seen in the ethanol
extracts Fig. 5. The wavelength at which
absorbance is the highest is called the lambda max
(λmax), and it is characteristic of particular groups of
constituents of the extract. This pattern over a
wavelength range can be used for substantiating the
identification of crude medicinal plants since this
value can be fairly constant and only varying with a
change in composition of the plant material. Hence
the UV absorption spectrum over the aqueous
Bekoe et al., IJPSR, 2021; Vol. 12(1): 515-523. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 521
extract from 200 to 600 nm and λmax of 3.8 at 233
nm and 2.7 at 280 nm can be used to authenticate
samples of K. senegalensis. Authentication can be
further established by observing the slight shifts in
absorbance in dilute acid and in the dilute base, as
seen in Fig. 4. Similar patterns can also be
observed with the ethanol extract.
Distilled water – 1 in 10 dilution Dil. NaOH – 1 in 10 dilution Dil. HCl – 1 in 10 dilution
Methanol – 1 in 10 dilution Methanolic NaOH – 1 in 10 dilution Methanolic HCl – 1 in 10 dilution
FIG. 4: UV SPECTRA OF AQUEOUS EXTRACT OF K. SENEGALENSIS IN VARIOUS SOLVENTS
Distilled water – 1 in 10 dilution Dil. NaOH – 1 in 10 dilution Dil. HCl – 1 in 10 dilution
Methanol – 1 in 10 dilution Methanolic NaOH – 1 in 10 dilution Methanolic HCl – 1 in 10 dilution
FIG. 5: UV SPECTRA OF ETHANOL EXTRACT OF K. SENEGALENSIS IN VARIOUS SOLVENTS
Bekoe et al., IJPSR, 2021; Vol. 12(1): 515-523. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 522
Heavy Metal Content: When the powdered stem
bark of K. senegalensis was analyzed, it showed
traces of heavy metals such as Cadmium (Cd),
Lead (Pb), etc. (refer to Table 7), but the levels
were within recommended limits 40.
TABLE 7: HEAVY METAL CONTENT
Metal
K. senegalensis (µg/kg)
Cadmium (Cd)
0.0029
Lead (Pb)
0.0085
Iron (Fe)
0.0466
Zinc (Zn)
0.0009
Arsenic (As)
-
Microbiological and Mutagenic Analysis:
Analysis for aerobic bacteria and fungi put an
estimated count of aerobic bacterial and fungal
cells at both 104 cells per gram of dried plant
material. It is recommended that, for plants
materials to which boiling water would be added,
such as K. senegalensis, aerobic bacteria and fungi
should be less than ≤ 105 and ≤ 104 41. Hence the
microbial counts of this plant material were within
range. Precaution would need to be exercised when
processing such materials so that the microbial
counts do not exceed the recommended value
because this could cause degradation of the product
and also reduce its efficacy, safety, and quality. K.
senegalensis exhibited weak mutagenicity in the
Ames test.
CONCLUSION: The botanical, chemical, bio-
logical, and physical parameters determined could
be pivotal in establishing the quality of crude plant
materials of K. senegalensis by such available
methods before they are used as medicines.
ACKNOWLEDGEMENT: The authors will like
to thank Miss Hannah Amponsah and Mr. Francis
Setsofia of the Department of Pharmacognosy and
Herbal Medicine and also Mr. Clement Sasu and
Mrs. Akosua Okraku of the Department of
Pharmaceutics and Microbiology, all of the
University of Ghana, for their technical assistance
when performing the various experiments.
CONFLICTS OF INTEREST: The authors
declare no conflict of interest.
REFERENCES:
1. Palhares RM, Goncalves Drummond M, Dos Santos Alves
Figueiredo Brasil B, Pereira Cosenza G, das Gracas Lins
Brandao M and Oliveira G: Medicinal plants
recommended by the world health organization: DNA
barcode identification associated with chemical analyses
guarantees their quality. PLoS One 2015; 10(5): e0127866.
2. Welz AN, Emberger-Klein A and Menrad K: Why people
use herbal medicine: insights from a focus-group study in
Germany. BMC Complementary and Alternative Medicine
2018; 18(1): 92.
3. Shakya AK: Medicinal plants:Future source of new drugs.
International Journal of Herbal Medicine 2016; 4(4): 59-
64.
4. WHO: Traditional Medicine Fact Sheet No 134 http://
wwwwhoint/mediacentre/factsheets/2003/fs134/en/,
accessed 7 March 2016.
5. Boadu AA and Asase A: Documentation of herbal
medicines used for the treatment and management of
human diseases by some communities in Southern Ghana.
Evidence-Based Complementary and Alternative Medicine
2017; 2017: 1-12.
6. Abdel-Aziz SM, Aeron A and Kahil TA: Health Benefits
and Possible Risks of Herbal Medicine. Microbes in Food
and Health 2016: 97-116.
7. Parveen A, Parveen B, Parveen R and Ahmad S:
Challenges and guidelines for clinical trial of herbal drugs.
Journal of Pharmacy and Bioallied Sciences 2015; 7(4):
32933.
8. Van Andel T, Myren B and van Onselen S: Ghana's Herbal
Market. Journal of Ethnopharmacology 2012; 140(2): 368-
78.
9. Oppong Bekoe E, Agyare C, Boakye YD, Baiden BM,
Asase A, Sarkodie J, Nettey H, Adu F, Otu PB and
Agyarkwa B: Ethnomedicinal survey and mutagenic
studies of plants used in Accra metropolis, Ghana. Journal
of Ethnopharmacology 2020; 248(112309): 1-12.
10. Rabadeaux C, Vallette L, Sirdaarta J, Davis C and Cock
IE: An examination of the Antimicrobial and Anticancer
Properties of Khaya senegalensis (Desr.) A. Juss. Bark E.
Pharmacognosy Journal 2017; 9(4): 504-18.
11. Orwa C, Mutua A, Kindt R, Jamnadass R and Anthony S:
In: Agroforestree Database: a tree reference and selection
guide version 40 2009: 1-5.
12. Onu A, Saidu Y, Ladan MJ, Bilbis LS, Aliero AA and
Sahabi SM: Effect of aqueous stem bark extract of Khaya
senegalensis on some biochemical, Haematological, and
Histopathological Parameters of Rats. Journal of
Toxicology 2013; 1-9.
13. Shayoub MElA, Kabbashi AS, Asma SMA, Mahmoud N,
Elhassan AM, Ahmed MA, Dawoud ADH, Jawad ASS
and Osman HM: Anti-malarial activity of Khaya
senegalensis. Indo American Journal of Pharmaceutical
Research 2016: 1-6.
14. Abdel-Wareth AA, Hammad S and Ahmed H: Effects of
Khaya senegalensis leaves on performance, carcass traits,
hemtological and biochemical parameters in rabbits.
EXCLI Journal 2014; 13: 502-12.
15. Zhang H, Tan J, Vanderveer D, Wang X, MJ W and Chen
F: Khayanolides from African mahogany Khaya
senegalensis (Meliaceae): A revision. Phytochemistry
2009; 70(2): 294-99.
16. Onu A, Saidu Y, Ladan M, Bilbis L, Aliero A and Sahabi
S: Effect of aqueous stem bark extract of Khaya
senegalensis on Some biochemical, haematological, and
histopathological parameters of rats. J Toxicol 2013; 9.
17. Adewuyi A, Otuechere CA, Adebayo OL, Anazodo C and
Pereira FV: Renal toxicological evaluations of sulphonated
nanocellulose from Khaya sengalensis seed in Wistar rats.
Chemico-Biological Interactions 2018; 284: 56-68.
18. Boffa J. Agroforestry parklands in sub-Saharan Africa.
FAO, Rome. Conservation Guide No. 34; 1999.
Bekoe et al., IJPSR, 2021; Vol. 12(1): 515-523. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 523
19. Idu M, Erhabor J, Oghale O and Obayagbona N:
Antimicrobial qualities, phytochemistry and micro-
nutritional content of Khaya senegalensis (Desr.) A. Juss
seed oil. The Journal of Phytopharmacology 2014; 3(2):
95-101.
20. Abdullah R, Diaz L, Wesseling S and Rietjens I: Risk
assessment of plant food supplements and other herbal
products containing aristolochic acids using the margin of
exposure (MOE) approach. Food Additives &
Contaminants: Part A 2000; 32(2): 135-44.
21. Hoang M, Chen C, Sidorenko V, He J, Dickman K, Yun
B, Moriya M, Niknafs N, Douville C and Karchin R:
Mutational signature of aristolochic acid exposure as
revealed by whole-exome sequencing. Science
Translational Medicine 2013; 5(197): 197-02.
22. Vanherweghem JL, Depierreux M, Tielemans C,
Abramowicz D, Dratwa M, Jadoul M, Richard C,
Vandervelde D, Verbeelen D and Vanhaelen-Fastre R:
Rapidly progressive interstitial renal fibrosis in young
women: association with slimming regimen including
Chinese herbs. Lancet 1993; 341(8842): 387-91.
23. Stiborova M, Arlt VM and Schmeiser HH: DNA adducts
formed by aristolochic acid are unique biomarkers of
exposure and explain the initiation phase of upper
urothelial cancer. International Journal of Molecular
Sciences 2017; 18(10): 2144.
24. Jadot I, Declèves A, Nortier J and Caron N: An integrated
view of aristolochic acid nephropathy: update of the
literature. Int J of Molecular Sciences 2017; 18(2): 297.
25. Dar RA, Shahnawaz M and Qazi PH: General overview of
medicinal plants: A review. The Journal of Phyto-
pharmacology 2017(6): 349-51.
26. Cardoso CR, de Syllos Colus IM, Bernardi CC,
Sannomiya M, Vilegas W and Varanda EA: Mutagenic
activity promoted by amentoflavone and methanolic
extract of Byrsonima crassa Niedenzu. Toxicology 2006;
225(1): 55-63.
27. Ghazali A, Abdullah R, Ramli N, Rajab N, Ahmad-Kamal
M and Yahya N: Mutagenic and antimutagenic activities
of Mitragyna speciosa Korth extract using Ames test.
Journal of Medicinal Plants Research 2011; 5(5): 1345-48.
28. McCann J, Ames BN: Detection of carcinogens as
mutagens in the Salmonella/microsome test: assay of 300
chemicals: discussion. Proceedings of the National
Academy of Sciences of the United States of America
1976; 73(3): 950-54.
29. Sponchiado G, Adam ML, Silva CD, Solely BS, Mello-
Sampayo C, Correl CJ and Otuki MI: Quantitative
genotoxicity assays for analysis of medicinal plants: A
systematic review. Journal of Ethnopharmacology 2016;
178: 289-96.
30. Guan Y, Wang X, Wong M, Sun G, An T, Guo J and
Zhang G: Evaluation of Genotoxic and Mutagenic Activity
of Organic Extracts from Drinking Water Sources. PLoS
One 2017; 12(1): e0170454.
31. Liu L, Chen L, Floehr T, Xiao H, Bluhm K, Hollert H and
Wu L: Assessment of the Mutagenicity of Sediments from
Yangtze River Estuary Using Salmonella Typhimurium/
Microsome Assay. PLoS One 2015; 10(11): e0143522.
32. Rodriguez E, Piccini C, Sosa V and Zunino P: The use of
the Ames test as a tool for addressing problem-based
learning in the microbiology lab. Journal of Microbiology
and Biology Education 2012; 13(2): 175-77.
33. WHO: Updated edition of Quality control methods for
medicinal plant materials, 1998; 2011.
34. Harborne JB: Phytochemical methods. In., edn. London:
Chapman and Hall Publications; 1992.
35. Khandelwal K: Practical Pharmacognosy Techniques and
Experiments. In., edn. New Delhi: Nirali Prakashan 2002.
36. Oppong Bekoe E, Dodoo KB, Kitcher C, Gordon A,
Frimpong-Manso S and Schwinger G: Pharmacognostic
Characteristics and Mutagenic Studies of Alstonia boonei
De Wild. Research Journal of Pharmacognosy 2020; 7(7):
7-15.
37. Ranjith D: Fluorescence analysis and extractive values of
herbal formulations used for wound healing activity in
animals. Journal of Medicinal Plants Studies 2018; 6(2):
189-92.
38. Ames B, McCann J, Yamasaki E: Methods for detecting
carcinogens and mutagens with the Salmonella/
mammalian-microsome mutagenicity test. Mutation
Research/Environmental Mutagenesis and Related
Subjects 1975; 31(6): 347-64.
39. Gilbert RI: The analysis of fluctuation tests. Mutation
Research 1980; 283: 289.
40. WHO: WHO guidelines for assessing quality of herbal
medicines with reference to contaminants and residues:
WHO Press, World Health Organization, 20 Avenue
Appia, 1211 Geneva 27, Switzerland; 2007.
41. British Pharmacopoeia. Appendix XVI (A – D). In: British
Pharmacopoeia, Volume IV. Wielka Brytania, Medicines
Commission 2004; 331-51.
All © 2013 are reserved by the International Journal of Pharmaceutical Sciences and Research. This Journal licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License.
This article can be downloaded to Android OS based mobile. Scan QR Code using Code/Bar Scanner from your mobile. (Scanners are available on Google
Playstore)
How to cite this article:
Bekoe EO, Kitcher C, Sarkodie JA, Dodoo KB, Frimpong-Manso S and Toafode MC: Evaluating pharmacognostic parameters and
mutagenic activity of Khaya senegalensis (Desr.) A. Juss. Int J Pharm Sci & Res 2021; 12(1): 515-23. doi: 10.13040/IJPSR.0975-
8232.12(1).515-23.
