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Chemical Analyses, Antimicrobial and Antioxidant Activities of Extracts from Cola nitida Seed

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Journal of Exploratory Research in Pharmacology
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Background and Objectives Medicinal plants are the richest, cheapest and most readily available source of drugs, nutraceuticals and food supplements. Pharmaceutical industries still rely largely on medicinal plants for intermediates due to their chemical diversities. This study, therefore, investigated the chemical constituents, thermal decomposition products and biological activities of extract from seeds of Cola nitida (the ‘kola nut’). Methods The pulverized seed was sequentially extracted with dichloromethane and methanol CH3OH. The extracts were analysed directly by Fourier Transform Infra-Red, electrospray ionization mass spectrometer and as fatty acid methyl ester and trimethylsilyl derivatives by gas chromatography-mass spectrometry (GC-MS). The CH3OH extract was analysed by high-performance liquid chromatography for sugars. The intact and extracted seed biomasses were analysed directly by pyrolysis GC-MS. For isolation of chemicals and assessment of biological activity, a large scale CH3OH extraction was performed and the extract partitioned with n-hexane, ethyl acetate (EtOAc) and butanol. Fractionation was done using various chromatographic techniques. Antimicrobial and antioxidant activities of the extract, fractions and isolated caffeine were respectively determined by the methods of agar-well diffusion and 2,2-diphenyl-1-picrylhydrazyl radical scavenging. Results Caffeine and hexadecanoic acid were isolated from the EtOAc fraction while theobromine, caffeine, catechins, procyanidins, proanthocyanidins, sugars, fatty acids, alcohols and sterols were identified in the extracts. Multitude (62) biomass degradation products were identified in pyrolysed seed samples. The extract and fractions showed varying activities against most of the tested microbes, except against Shigella species, for which neither the extract nor fractions elicited any response. The butanol fraction exhibited the highest antioxidant activity. Conclusions This report gives insight into the chemical constituents in Cola nitida seed, details the thermal decomposition constituents and establishes the antimicrobial and antioxidant activities of the seed extract and fractions, thereby contributing to the knowledge on the chemistry and pharmacology of the genus.
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Journal of Exploratory Research in Pharmacology 2017 vol. 2 | 67–77
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Original Article
Chemical Analyses, Antimicrobial and Antioxidant Activities of
Extracts from Cola nitida Seed
Julius K. Adesanwo1*, Seun B. Ogundele1, David A. Akinpelu2 and Armando G. McDonald3
1Department of Chemistry, Obafemi Awolowo University, Ile-Ife, Nigeria; 2Department of Microbiology, Obafemi Awolowo University,
Ile-Ife, Nigeria; 3Department of Forestry, Rangeland and Fire Sciences, University of Idaho, Moscow, ID 83843, USA
Abstract
Background and Objecves: Medicinal plants are the richest, cheapest and most readily available source of drugs,
nutraceucals and food supplements. Pharmaceucal industries sll rely largely on medicinal plants for interme-
diates due to their chemical diversies. This study, therefore, invesgated the chemical constuents, thermal
decomposion products and biological acvies of extract from seeds of Cola nida (the ‘kola nut’).
Methods: The pulverized seed was sequenally extracted with dichloromethane and methanol CH3OH. The ex-
tracts were analysed directly by Fourier Transform Infra-Red, electrospray ionizaon mass spectrometer and as
fay acid methyl ester and trimethylsilyl derivaves by gas chromatography-mass spectrometry (GC-MS). The
CH3OH extract was analysed by high-performance liquid chromatography for sugars. The intact and extracted
seed biomasses were analysed directly by pyrolysis GC-MS. For isolaon of chemicals and assessment of biologi-
cal acvity, a large scale CH3OH extracon was performed and the extract paroned with n-hexane, ethyl ac-
etate (EtOAc) and butanol. Fraconaon was done using various chromatographic techniques. Anmicrobial and
anoxidant acvies of the extract, fracons and isolated caeine were respecvely determined by the methods
of agar-well diusion and 2,2-diphenyl-1-picrylhydrazyl radical scavenging.
Results: Caeine and hexadecanoic acid were isolated from the EtOAc fracon while theobromine, caeine, cat-
echins, procyanidins, proanthocyanidins, sugars, fay acids, alcohols and sterols were idened in the extracts.
Multude (62) biomass degradaon products were idened in pyrolysed seed samples. The extract and frac-
ons showed varying acvies against most of the tested microbes, except against Shigella species, for which
neither the extract nor fracons elicited any response. The butanol fracon exhibited the highest anoxidant
acvity.
Conclusions: This report gives insight into the chemi-
cal constuents in Cola nida seed, details the ther-
mal decomposion constuents and establishes the
anmicrobial and anoxidant acvies of the seed
extract and fracons, thereby contribung to the
knowledge on the chemistry and pharmacology of
the genus.
Keywords: Cola nitida; Caffeine; n-Hexadecanoic acid; Catechin; Antimicrobial;
Antioxidant; GC-MS; ESI-MS.
Abbreviations: AGC, Acelerated Gradient Chromatography; 13CNMR, Carbon
13 Nuclear magnetic resonance spectroscopy; CND, Cola nitida dichloromethane
extract; CNM, Cola nitida methanol extract; IC50 value, concentration of samples
leading to 50% reduction of initial DPPH radical concentration; CDCl3, deuterated
chloroform; DCM or CH2Cl2, dichloromethane; DPPH, 2,2-diphenyl-1-picrylhydra-
zyl; ESI-MS, electrospray ionization mass spectrometer; EFAs, essential fatty acids;
EtOAc, ethyl acetate; FAME, fatty acid methyl ester; FAs, fatty acids; FTIR, fourier
transform infrared; GC-MS, gas chromatography-mass spectrometry; GRAM +ve,
Gram positive; GRAM −ve, Gram negative; HPLC, high-performance liquid chroma-
tography; IS, internal standard; CH3OH, methanol; 1HNMR, proton nuclear magnetic
resonance spectroscopy; Pyro GC-MS or Py GC-MS, pyrolysis gas chromatography-
mass spectrometry; Ag, silver; TLC, thin-layer chromatography; TMS, trimethylsilyl;
UV, Ultra- violet; ZnSe, Zinc Selenium.
Received: April 28, 2017; Revised: June 23, 2017; Accepted: June 28, 2017
*Correspondence to: Julius K. Adesanwo, Department of Chemistry, Obafemi
Awolowo University, Ile-Ife, Nigeria. Tel: +234-8030821561, E-mail: adesanwojk@
yahoo.com or julius08@oauife.edu.ng
How to cite this article: Adesanwo JK, Ogundele SB, Akinpelu DA, McDon-
ald AG. Chemical Analyses, Antimicrobial and Antioxidant Activities of Extracts
from Cola nitida Seed. J Explor Res Pharmacol 2017;2(3):67–77. doi: 10.14218/
JERP.2017.00015.
Introduction
Cola nitida (Vent) Schott et Endl, family Malyaceae, is a tree na-
tive to the rainforest of tropical West Africa. The seed or nut com-
monly called ‘kola nut’ is a popular stimulant in West Africa. It
is called “Obi gbanja” by Yorubas in western Nigeria, “Goro” by
Hausas in the north, and “Oji” by Ibos in the east.1 It is mainly used
for nutritional purpose, eaten across cultures in West Africa, and
DOI: 10.14218/JERP.2017.00015 | Volume 2 Issue 3, August 201768
Adesanwo JK.. et al: Medicinal activities of Cola nitida seed
J Explor Res Pharmacol
has great social, religious and medicinal importance as well.2 Kola
nut extracts are used in the manufacture of various non-alcoholic
beverages, soft drinks, wines, chocolate and sweets.3–5
Previous researchers have reported a number of secondary me-
tabolites from the kola nut, including caffeine, theobromin, cate-
chin, epicatechin, procyanidins and proanthocyanidins.6–8 Aliphat-
ic and heterocyclic amines in different species of cola have been
reported as well,9 and quinic, tannic and chlorogenic acids have
been found as present in the kola nut.10 Caffeine was the major
alkaloid identied in Cola seeds and was considered as one of the
signature compounds due to its concentration range.6
Knebel and Higler showed that fresh kola nut contained a glu-
coside named “kolanin”, reporting that it is readily hydrolysed or
split into glucose and caffeine in ripe or dried fruit.7 Kola nuts are
rich in xanthine alkaloids, such as theobromine, caffeine, kolatin
and kolanin. While caffeine stimulates the body, kolatin stimu-
lates the heart.11,12 These constituents (caffeine, theobromine, and
theophylline) in kola nut extract have been shown to contribute
to the anti-photodamage effect, including wrinkles, and to elicit
antioxidant and anti-aging activities, including suppression of UV-
induced erythema, and to decrease skin roughness and scaling by
topical or oral application.13
Nowadays, pathogenic microorganisms are developing resist-
ance to existing antibiotics at an alarming rate. Extract of the
leaves, root and stem bark of Cola nitida are extensively used in
folk medicine.6 Different parts of Cola nitida have been exploit-
ed for different biotechnological applications. For instance, cola
pods have been used for production of fructosyltransferase, an
important enzyme for the synthesis of fructooligosaccharides,14
and most recently for the biosynthesis of silver (Ag) nanoparticles
and silver-gold alloy nanoparticles for diverse biomedical appli-
cations,15–19 such as antimicrobial, antioxidant, anticoagulant and
thrombolytic activities.
Similarly, the seed and seed shell extracts of Cola nitida have
been used for green synthesis of Ag nanoparticles, showing pro-
found antibacterial activities.20 In Uganda, extracts of the seed and
leaves are used in treatment of sexual and erectile dysfunctions.21
In Nigeria, methanol extract of the seed is used against emesis and
migraine.22 The aqueous extract of the seed has also been used as
avouring in carbonated drinks.23 Kola nut extract also demon-
strates good protective property against red cell degradation.9
The aims of this study were to: (i) determine the composition of
kola nut dichloromethane (CH2Cl2) and CH3OH extracts by a com-
bination of electrospray ionization-mass spectrometry (ESI-MS)
and gas chromatography-mass spectrometry (GC-MS) analyses;
(ii) determine the antimicrobial and antioxidant activity of the ex-
tracts; and (iii) isolate the active principle component(s).
Experimental
General
All thin-layer chromatography (TLC) analyses were performed at
room temperature using pre-coated plates (silica gel 60 F254 0.2
mm; Merck). Detection of spots was achieved by staining with
iodine crystals and exposure to ultraviolet light (254 and 366
nm). Melting point determination was carried out using a Gallen-
kamp apparatus. Accelerated gradient chromatography (Baeck-
strom Separo AB) was carried out using silica gel (Kieselgel 60G
0.040–0.063 mm) and column chromatography using silica gel,
with a 60–200 mesh for fractionation. Proton Nuclear magnetic
resonance (1HNMR) spectroscopic data were recorded on a NMR
machine (Agilent Technologies) at 400 MHz and at 100 MHz for
13CNMR. Chemical shifts of signals were reported in parts per mil-
lion (ppm).
Collection of plant material
The plant material (kola nut) was collected at Alaro Farm Settle-
ment, Ile-Ife, identied and authenticated at the Department of
Pharmacognosy, Obafemi Awolowo University (Voucher Number
FPI 2052).
Sample preparation and extraction
The seeds were dried at 60 °C for 48 h, and milled to particle size
of 1 mm. Moisture content was determined in duplicate before
extraction. The plant material was Soxhlet extracted (40 g, in du-
plicate) rst with dichloromethane (DCM) (150 mL) for 24 h and
then with CH3OH (150 mL) for 48 h. The extracts were concen-
trated in vacuo at 40 °C to give yield of 0.77% and 17.62% for the
DCM and CH3OH extracts, respectively.
Fourier transform infrared (FTIR) spectroscopic analysis
The functional groups in the milled samples and extracts, and the
extracted biomasses were determined by FTIR spectroscopy using
a Nicolet iS5 spectrometer (ThermoScientic) using a ZnSe at-
tenuated total reection probe. Spectra were collected in duplicate.
The absorbance spectra were baseline-corrected and averaged us-
ing Omnic v9.0 software (ThermoScientic).24,25
ESI-MS experiment
The samples (about 1.0 mg each of DCM and CH3OH extracts, in
duplicate) were added to CH3OH (1 m:) and acetic acid (10 µL).
The mixture was subjected to sonication to ensure total dissolu-
tion. Electrospray mass spectrometric analyses were performed on
a 5989A device (Hewlett-Packard) equipped with an electrospray
interface 59987A. Nitrogen was used as nebulizing gas, at a pres-
sure of 50 psi and a temperature of 300 °C. Sample analysis was
performed by direct infusion in ESI-MS using a syringe pump
(Harvard Apparatus) at a ow-rate of 10 mL/min. Mass spectra
were acquired in scan mode detection, and ESI-MS conditions
were optimized using available standards.
GC-MS of fatty acid methyl ester (FAME) derivatives
Extracts (about 2.0 mg, in duplicate) were prepared by adding
a solution (2 mL) of CH3OH/H2SO4/CHCl3 (1.7:0.3:2.0 v/v) in
which the CHCl3 contained 1-naphthaleneacetic acid (100 µg/
mL) as an internal standard. The mixture was heated for 90 min
at 90 °C in a sealed vial. Water was added to the mixture and the
CHCl3 layer was removed, dried and transferred to a GC vial.
The prepared FAME derivatives were analysed by electron impact
ionization GC-MS on a Focus ISQ (ThermoScientic) with a ZB5
column (30 m × 0.25 mm; Phenomenex) and a temperature pro-
le of 40 °C (1 min) to 320 °C (10 min) at 5 °C/min. The eluted
compounds were identied with authentic standards (C12 to C20
fatty acids) and by spectral matching with the NIST 2008 spectral
library.
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Adesanwo JK.. et al: Medicinal activities of Cola nitida seed J Explor Res Pharmacol
GC-MS of extracts for trimethylsilyl (TMS) derivatives
Extracts (about 1.0 mg, in duplicate) were weighed in GC vials, to
which CH2Cl2 (1 mL) containing anthracene as an internal stand-
ard (IS; 50 µg/mL) was added. The samples were silylated with
addition of N,O-bis(trimethylsilyl)-triuoro-acetamide (BSTFA)
containing 1% trimethylchlorosilane (TMCS; 50 µL) and pyridine
(50 µL) and heated for 30 min at 70 °C or longer until the solu-
tion became clear. The prepared TMS derivatives were analysed by
GC-MS (as described above).
Analytical pyro (Py)-GC-MS of samples
Analytical Py-GC-MS was carried out on the milled sample before
and after extraction using a Pyrojector II (SGE Analytical Science)
at 500 °C in He coupled to a GC-MS (FOCUS-ISQ; ThermoScien-
tic) instrument operating in the electron impact ionization mode.
The compounds were separated on the ZB5-MS capillary column
(30 mm × 0.25 mm; Phenomenex) with temperature programmed
to be 50 °C to 3000 °C, at 5 °C min−1. The eluted compounds
were identied by their mass spectra, authentic standards, and with
NIST 2008 library matching.
High-performance liquid chromatography (HPLC) analysis
Sugars were quantied by HPLC using a Rezex ROA column (7.8
mm × 30 cm; Phenomenex) and a Waters HPLC (Waters Corp.)
equipped with differential refractive index detector (Shodex SE61;
Showa Denko America, Inc.), on elution with 0.01 M H2SO4 (0.5
mL/min) at 65 °C. The CH3OH extract (10 mg) was dissolved in
0.01 M H2SO4 (5 mL), centrifuged and the supernatant ltered
(0.45 µm). Data was acquired and analysed using the N2000 chro-
matography software (Science Technology Inc.). The sugar con-
tents were determined from peak area using the external standard
method with standard sugars (glucose, fructose, xylose, myo-ino-
sitol, sucrose, maltose and turanose).
Isolation of chemical compounds
The seeds (2 kg) were extracted with CH3OH and concentrated
in vacuo to obtain CH3OH extract (80 g, 4% yield). The crude
extract was suspended in water and partitioned with n-hexane,
ethyl acetate (EtOAc) and n-butanol to give respective fractions.
The EtOAc fraction was subjected to Acelerated Gradient Chro-
matography (AGC) fractionation. Fractions 24 through 76 were
bulked together and separated by column chromatography (silica
gel 60–200 mesh) and elution was monitored by TLC. Fractions 36
through 67 were evaporated to yield a white crystalline solid (caf-
feine) with melting point of 230–233 °C. Pooled AGC fractions
8 through 23 were concentrated and a yellow viscous solid was
obtained, which was further puried by column chromatography,
yielding a colourless solid (hexadecanoic acid) with minor impuri-
ties as accessed by TLC. Both compounds were characterized by
NMR spectroscopy (400 MHz; Agilent).
Antimicrobial assay
The antimicrobial activity of CH3OH extract, solvent fractions and
isolated caffeine were determined using the agar-well diffusion
method.26,27 The bacteria were grown in a nutrient broth before
use, while the fungi were grown on potato dextrose agar medi-
um until they sporulated, at which time they were harvested. The
standardized bacteria suspension was spread on Muller-Hinton
agar and allowed to set. Wells were then bored with a 6-mm borer
and lled with the respective sample solutions at 10 mg/mL, with
ampicillin and nystatin added at 1 mg/mL, which was followed by
incubation at 37 °C for 24 hrs. The fungal plates were incubated at
25 °C for 96 hrs.
2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging an-
tioxidant assay
Quantitative antioxidant activity was determined spectropho-
tometrically as described.28 Reactions were carried out in test
tubes, and each of the solvent fractions and CH3OH extracts were
tested at varying concentrations (µg/mL). Initial stock solutions
of 1 mg/mL were prepared for the various plant extracts. The
following nal concentrations were prepared based on the pre-
liminary qualitative TLC antioxidant screening: 60, 30, 15, 7.5,
3.75 and 1.875 µg/mL for the crude extract; 50, 25, 12.5, 6.25
and 3.13 µg/mL for n-butanol fraction; 30, 15, 7.5, 3.75 and 1.88
µg/mL for the EtOAc fraction; and 2000, 1000, 500, 250, 125 and
62.5 µg/mL for the n-hexane extract and caffeine from the stock
solution. A 0.1 mM DPPH radical solution in CH3OH (1 mL) was
added to 1 mL of the concentration series for each sample tested,
in triplicate, and allowed to react at room temperature in the dark
for 30 min. The negative control was prepared by adding 0.1 mM
DPPH radical solution (1 mL) to 1 mL of methanol, in triplicate,
and absorbance was measured at 517 nm. The percentage anti-
oxidant activity (%AA) values of test samples were calculated
from the absorbance using the formula:% AA={Xcontrol−Xtest
Xcontrol}×100where Xcontrol is the absorbance of the negative
control, Xtest is the absorbance of test samples concentrations.
Ascorbic acid (vitamin C) was used as the standard antioxidant
Fig. 1. Fourier transform infrared spectra of Cola nida seed. (A) Pow-
dered unextracted material, (B) Cola nida dichloromethane (CND) ex-
tract, (C) Cola nida methanol (CNM) extract, (D) Extracted biomass.
DOI: 10.14218/JERP.2017.00015 | Volume 2 Issue 3, August 201770
Adesanwo JK.. et al: Medicinal activities of Cola nitida seed
J Explor Res Pharmacol
agent.
The IC50 value (i.e. the concentration of the test samples leading
to 50% reduction of the initial DPPH radical concentration) was
calculated from the separate linear regression of plots of the mean
percentage of antioxidant activity against the concentration of the
test samples in µg/mL.
Results and discussion
Extraction techniques
Soxhlet extraction of the kola nut with CH2Cl2 and CH3OH af-
forded yields of 0.77% and 17.6%, respectively. In comparison,
large-scale cold extraction of the kola nut afforded a low yield of
4%. Therefore, only the Soxhlet extracts were analysed in detail
by a combination of GC-MS and ESI-MS. However, the batch
CH3OH extract was used for antioxidant and antimicrobial assess-
ment.
FTIR spectroscopic analysis
FTIR spectroscopy was used to investigate the functional groups
in the sample and extracts. The spectra (Fig. 1) showed the pres-
ence of strong O–H stretching vibration at 3340 cm−1 correspond-
ing to hydroxyl groups in all, but quite pronounced in Figure 1A,
C, and D. This band is less pronounced in Figure 1B, because the
extract contained a majority of low polar compounds, whereas
Figure 1C and D contained high polar phenolic compounds and
lignocellulose respectively and containing multiple H-bonding.
The absorption at just above 3000 cm−1 in Figure 1a is due to an
aromatic C–H stretch of polyphenols and aromatics; it is virtu-
ally absent in Figure 1b but noticeable in Figure 1C and D. Simi-
larly, the substituted aromatic C–H bend band at 600–900 cm−1
is almost absent in Figure 1b but very conspicuous in Figure 1A,
C and D, indicating a low proportion of aromatics in the CND
extract.
On the other hand, the C–H sp3 stretching vibration, just below
3000 cm−1 and characteristic of long chain fatty acids [–(CH2)
n–CH3], is more conspicuously prominent in Figure 1B than in
Figure 1C and D. Carbonyl stretching absorption of carboxylic
acid aldehydes and ketones is very low in Figure 1D, as most car-
bonyl compounds had already been removed through the process
of extraction. The absorption band at 1000–1200 cm−1 that was
due to C–O stretch of esters (glycosides and cellulose) was most
intense in Figure 1C and D but low in Figure 1B. These data show
that FTIR can be used to monitor and assess the effectiveness of
the extraction process, apart from facilitating quantitative evalu-
Fig. 2. Electrospray ionizaon mass spectra of Cola nida extracts posive mode. (A) Cola nida methanol extract, (B) Cola nida dichloromethane extract.
DOI: 10.14218/JERP.2017.00015 | Volume 2 Issue 3, August 2017 71
Adesanwo JK.. et al: Medicinal activities of Cola nitida seed J Explor Res Pharmacol
ations.
Electrospray mass spectrometric analysis of extracts
ESI-MS is a powerful analytical tool to rapidly analyse extracts
and isolated fractions without chromatographic separation.29–31
Moreover, ESI-MS makes it possible to discriminate between var-
ious avonoid classes, provide information on the glycosylation
position and characterize saponins.32–34 Therefore, ESI-MS was
employed to directly analyse the CH2Cl2 and CH3OH extracts in
positive (Fig. 2) and negative (Fig. 3) ion modes.
The positive ion ESI-MS for both extracts showed the larg-
est peak at m/z 195 (100% intensity), which was due to caffeine
([M+H]+). In the MeOH extract (Fig. 2A), the tentative peak assign-
ments are as follows: 181 ([M+H]+, theobromine); 295 (unknown);
and 391 (unknown). For the CH2Cl2 extract (Fig. 2B), the tentative
peak assignments are: 193 ([M+H]+, quinic acid); 313 ([M+Na]+,
catechin), 355 ([M+H]+, chlorogenic acid); and m/z 281, 294, 377
and 426 (unknowns). The negative ion ESI-MS for both extracts
(Fig. 3) showed a multitude of peaks. For the CH3OH extract (Fig.
3A), the tentative assignments are: m/z 289([M-H], catechin);
m/z 577 and 578 (procyanidins B1, B2 [oligomeric catechins] and
naringin avonoids); and m/z 1153 and 1154 (proanthocyanidins).
There were many unidentied peaks. Niemenak et al.6 also reported
that HPLC of Cola nitida extract had 11 unidentied compounds.
GC-MS of extracts of FAME derivatives
It is well known that fatty acids (FAs), especially the essential fat-
ty acids (EFAs), are important for optimal functioning of the body.
They regulate such bodily functions as heart rate, blood pressure,
blood clotting and fertility. They also participate in immune sys-
tem inammation against harmful waste products. Balance of
EFAs is important for good health and normal development of
humans.35,36 FAs occur widely in natural fats and dietary oils and
they play important roles as nutritious substances and metabolites
in living organisms.37 With these important biological activities of
FAs, those present in the extracts as free or glycerides were con-
Fig. 3. Electrospray ionizaon mass spectra of Cola nida extracts negave mode. (a) Cola nida methanol extract, (b) Cola nida dichloromethane extract.
DOI: 10.14218/JERP.2017.00015 | Volume 2 Issue 3, August 201772
Adesanwo JK.. et al: Medicinal activities of Cola nitida seed
J Explor Res Pharmacol
verted to FAME derivatives for identication and quantication.
The CH2Cl2 and CH3OH extracts were derivatised into FAME
and analysed by GC-MS. The CH2Cl2 extract contained FAs rang-
ing from C16 to C19. Palmitic acid (C16: 0.447 mg/g), linoleic
acid (C18: 0.198 mg/g), oleic acid (C18: 0.553 mg/g), stearic acid
(C18: 0.034mg/g), 8,9-methylene-heptadec-8-oic acid (C18: 0.112
mg/g), 10,12-octadecadienoic acid (C18: 0.040 mg/g), 8-oxohexa-
decanoic acid (C16: 0.275 mg/g) and 9,10-methylene-octadec-
9-oic acid (C19: 0.032 mg/g) were the major FAs detected. The
FAs constitute about 73% of the extract. Furthermore, caffeine,
fatty alcohols and sterols were also detected (Table 1). The CH3OH
extract did not contain any FAs, only caffeine.
GC-MS of extracts of TMS derivatives
Trimethylsilylating reagent is routinely used to derivatise rather
non-volatile compounds, such as certain alcohols, phenols, or car-
boxylic acids, by substituting a TMS group for a hydrogen in the
hydroxyl groups on the compounds. Both extracts were deriva-
tised to TMS ethers/esters to conduct the analyses for non-polar
and polar components using GC-MS. The results for the CH2Cl2
and CH3OH extracts as their TMS derivatives are given in Tables
2 and 3.
Caffeine (17.9% and 3.85% in CH3OH and CH2Cl2 extracts, re-
spectively) was predominant, as observed by Niemenak et al.6 Cat-
echin was the dominant avonoid in the kola seed. Other identied
constituents of CH3OH extract included: sugars (45%); catechin
(13%); malic acid and glycerol (Table 2). The presence of glucose,
fructose and sucrose in the CH3OH extract was detected by both
GC-MS and HPLC analyses. The CH2Cl2 extract consist mainly of
FFAs (Table 3), and this observation is consistent with the results
of FAME analysis. Other compounds found in this extract were
alkaloids and sterols.
Table 2. GC-MS of Cola nida CH3OH extracts of TMS derivaves
Compound Molecular formula Class Molecular weight Retenon me (min) % Extract
Glycerol TMS C12H32O3Si3Alcohol 308 18.11 0.2
Malic acid TMS C13H30O5Si3Acid 350 23.65 1.5
Anthracene (IS) C14H10 178 30.14
Fructose TMS5C19H46O6Si4Sugar 540 30.70 2.2
Caeine C8H10N4O2Alkaloid 194 31.23 17.9
Glucose-TMS-5 C21H52O6Si5Sugar 540 32.77 0.7
D-Turanose-TMS-7 C33H78O11Si7Sugar 846 45.65 2.9
Sucrose-TMS8C36H86O11Si8Sugar 918 45.75 39.5
Catechin-TMS-5 C30H54O6Si5Flavonoid 650 48.74 12.9
Table 1. GC–MS FAME derivaves of CND extract
S/No Compound Molecular formula Class Molecular
weight
Retenon
me (min) % Extract
1 Naphthalene acec acid (IS) C13H12O2200 27.34
2 Caeine C8H10N4O2Alkaloid 194 30.43 1.7
3Palmic acid C17H34O2FA 270 31.83 19.4
4Linoleic acid C19H34O2FA 294 35 8.6
5 Oleic acid C19H36O2FA 296 35.11 24.0
6 Stearic acid C19H38O2FA 298 35.6 1.5
7 8,9-Methylene-8-heptadecenoic acid C19H34O2FA 294 36.24 4.9
810,12-Octadecadienoic acid C19H34O2FA 294 36.39 1.7
9 7-(Tetrahydro-2H-pyran-2-yloxy)-2-octyn-1-ol C13H22O3Alcohol 226 36.63 1.3
10 2-Octylcyclopropene-1-heptanol C18H34O Alcohol 266 36.79 1.7
11 8-Oxohexadecanoic acid C17H32O3FA 284 37.25 12.0
12 Tetrahydropyran-2-yl ether of 7-dodecynol C17H30O2Alcohol 266 37.46 1.2
13 9,10-Methylene-9-octadecenoic acid C20H36O2FA 308 37.99 1.4
14 1-Ethyl-(1,1-dimethylethyl)-methoxycyclohexan-1-ol C13H26O2Alcohol 214 38.97 3.2
15 Sgmastan-3,5-diene C29H48 Sterol 396 50.12 0.2
16 Sitosterol acetate C31H52O2Sterol 456 52.16 0.7
17 Sitosterol C29H50OSterol 414 52.86 0.6
DOI: 10.14218/JERP.2017.00015 | Volume 2 Issue 3, August 2017 73
Adesanwo JK.. et al: Medicinal activities of Cola nitida seed J Explor Res Pharmacol
Analytical Py-GC-MS of samples
Direct analysis of the kola nut and extracted nut were performed
by analytical Py-GC-MS. The identities of the products are given
in Table 4. The main compounds in the kola nut were caffeine
(22.5%), CO2 (12.8%), methyl acetate (6.3%), acetic formic anhy-
dride (5.9%), levoglucosan (5.4%), N-methyl ethylamine (4.9%),
1,2-cyclopentanedione (3.4%) and pyrocatechol (3.0%). Aliphatic
and heterocyclic (pyrolidine, alstonine and xanthenes) amines were
identied. This nding is in accordance with the report by Atawodi
et al.9 Py-GC-MS of the extracted kola nut showed the presence
of 22 compounds (Table 4). The main compounds identied were:
CO2 (17%), acetol (11%), 1,2-benzenediol (11%), 1,2-cyclopentan-
edione (10%), acetic acid (7%), butylamine (5%) and caffeine (5%).
It is noteworthy that alkanes, alkenes ethers and steroids/sterols
were not identied by the Py-GC-MS of the extracted biomass;
this may be explained as due to exhaustive removal of the relative-
ly low polar compounds during the process of extraction. Analyti-
cal pyrolysis experiment showed caffeine as the major constituent
of Cola nitida. Maltol, a naturally occurring organic compound,
is primarily used as a avour enhancer.10 Its presence in the Cola
nitida seed may be contributory to the application of the seed in the
manufacture of soft drinks.3–5
Structure elucidation
Compound 1 (caffeine) was obtained as a white crystalline solid,
with a melting point of 230–233 °C. 1HNMR (400 MHz, CDCl3,
δ ppm): 7.46 (1Hs, 8H), 3.95, 3.54, 3.36 (s, 3H at 1,3,7). 13CNMR
(100 MHz, CDCl3, δ ppm): 155.3 and 151.6 (C6 and C2 respec-
tively), 148.6, 141.5, 107.5 (olenic C8, C4 and C5), 33.5, 29.7
and 27.9 (methyl groups). These correlate with the literature data
for caffeine.
Compound 2 (n-Hexadecanoic acid): 1HNMR (400 MHz, CDCl3,
δ ppm): 2.24–1.21 (CH2 protons), 0.78 (CH3 protons). 13CNMR
(100 MHz, CDCl3, δ ppm): 179 (C=O); 34.15–22.64 (CH2 carbon
atoms); 15.02 CH3 group.
Antimicrobial activity
The kola nut CH3OH extract and fractions, and isolated caffeine
showed varying degrees of inhibitory activities against the tested
bacterial and fungal strains (Fig. 4 and Supplementary Table S1).
Their activity was more pronounced against Gram-positive bacte-
ria than Gram-negative bacteria. This could be a result of the mor-
phological differences between these microorganisms. The Gram-
positive bacteria have an outer peptidoglycan layer, which is not an
effective permeability barrier, making these microorganisms more
susceptible to the compounds under investigation; meanwhile, the
Gram-negative bacteria have an outer phospholipidic membrane
that contains LPSs, making the cell wall of these microorganisms
impermeable.38–40.
These microorganisms are implicated in the pathogenesis of hu-
man infections. The result obtained showed the EtOAc fraction
as having the highest antimicrobial activity against most of the
organisms compared to other fractions. However, the EtOAc frac-
tion was not active against Pseudomonas Spp., Clostridium sporo-
genes, Corynebacterium pyogenes, Shigella Spp. and Candida
albicans. The activity of the extract indicated CH3OH as a good
solvent for preparation of extracts for antimicrobial assay.41,42 The
antimicrobial activity of the CH3OH extract can be attributed to
synergistic effect of compounds in the extract. Isolated caffeine
(compound 1) demonstrated higher activity than n-butanol against
Bacillus Spp., Escherichia coli and Aspergillus niger. Extract and
fractions did not show activity against Shigella Spp. Caffeine dem-
onstrated appreciable antimicrobial activity against Bacillus cere-
us, Escherichia coli, Pseudomonas vulgaris and the fungi.
Antioxidant activity
The DPPH radical antioxidant activity showed the ability of the
kola nut CH3OH extracts and fractions, and isolated caffeine to
reduce DPPH radicals through the transfer of acidic labile protons
by a free radical mechanism. None of the extracts compares sig-
nicantly with that of ascorbic acid. However, the EtOAc and bu-
tanol fractions exhibited good DPPH antioxidant activity. The IC50
values decreased in this order: ascorbic acid (3.2 ± 0.05 µg/mL),
butanol extract (9.8 ± 0.5 µg/mL), EtOAc extract (15.1 ± 0.7 µg/
mL), CH3OH extract (22.7 ± 1.7 µg/mL), hexane extract (321 ± 7
µg/mL) and caffeine (1370 ± 19 µg/mL).
Future research direction
A number of chemical compounds have been reported from the
Table 3. GC-MS of Cola nida CH2Cl2 extracts of TMS derivaves
Compound Molecular formula Class Molecular weight Retenon me (min) % Extract
Nonanoic acid TMS C12H26O2Si FA 230 20.14 0.82
Octanedioic acid TMS C14H30O4Si2FA 318 28.27 0.26
Anthracene C14H10 178 30.14
Caeine C8H10N4O2Alkaloid 194 31.32 2.95
Palmic acid TMS C19H40O2Si FA 328 35.07 1.64
Linoleic acid TMS C21H40O2Si FA 352 37.94 5.95
Oleic acid TMS C21H42O2Si FA 354 38.11 1.89
Stearic acid TMS C21H44O2Si FA 356 38.58 0.14
Linolenic acid TMS C21H38O2Si FA 350 39.57 1.08
Sgmasterol TMS C32H56OSi Sterol 484 53.46 0.20
Sitosterol TMS C32H58OSi Sterol 486 54.13 2.05
DOI: 10.14218/JERP.2017.00015 | Volume 2 Issue 3, August 201774
Adesanwo JK.. et al: Medicinal activities of Cola nitida seed
J Explor Res Pharmacol
Table 4. Py-GC-MS of unextracted and extracted powdered Cola nida seed
Compound Molecular
formula
Molecular
weight
Retenon
me (min)
Kola
nut, %
Extracted
nut, %
CO2CO244 1.12 12.8 17.4
N-Methyl ethylamine C3H9N 59 1.30 4.9
Butylamine C4H11N 72 1.38 4.9
Acec acid C3H4O360 1.64 5.9 6.5
Pentanone C5H10O86 1.65 4.6
Acetol C3H6O274 1.97 6.3 11.3
Unknown 92 3.17 1.5
Butanedial C4H6O286 3.36 1.7 3.4
Methyl pyruvate C4H6O3102 3.49 1.7 1.1
Butanedial C4H6O286 3.63 2.7
Unknown C4H4O284 3.66 1.0 1.1
2-Oxo-3-cyclopentene-1-acetaldehyde? C7H8O2124 4.37 1.3 3.2
2-Furfuryl alcohol C5H6O298 4.90 2.2 4.4
Unknown C5H8O3116 5.16 1.1
2(5H)-Furanone C4H4O284 6.34 2.1 3.9
1,2-Cyclopentanedione C5H6O298 6.67 3.4 9.9
1-Methyl-1-cyclopenten-3-one C6H8O96 7.85 1.3
4-Methyl-5H-furan-2-one + Unknown C5H6O298 + 110 8.09 0.6
Phenol C6H6O94 8.28 XX
2 Hydroxy-3-methyl-2-cyclopenten-1-one C6H8O2112 9.54 1.4 5.4
2,3 Dimethyl-2-cyclopenten-1-one C7H10O110 9.85 0.3
Unknown 116 10.00 0.8
2-Methyl phenol C7H8O2108 10.40 0.7
3-Methyl-phenol C7H8O2108 10.99 0.7
Unknown 57? 11.51 1.9 0.9
3 Hydroxy-2-methyl-4H-pyran-4-one (Maltol) C6H6O3126 12.03 0.5 0.7
3-Ethyl-2-hydroxy-2-cyclopenten-1-one C7H10O2126 12.21 0.4 1.5
4-Hydroxy-3-methyl-(5H)-furanone or 3-methyl-2,4(3H,5H)-furandione C5H6O3114 12.33 0.5
Dihydro-2H-pyran-3(4H)-one + unknown C5H8O2100+128 13.16 0.8
Ethyl/dimethyl phenol + 3,5-dihydroxy-2-methyl-(4H)-pyran-4-one C11H18O7122+142 13.66 0.7
Benzene diol + 3,5 dihydroxy-2 methyl-4-pyrone C6H6O2 + C6H6O4110 + 142 14.13 0.7
5-Hydroxymethyldihydrofuran-2-one C5H8O3116 14.42 0.8
1,2-Benzenediol C6H6O2110 14.67 3.1 11.3
1,4:3,6-Dianhydro-hexose C6H8O4144 14.86 1.0 1.9
Coumaran C8H80120 15.13 0.5
5 Hydroxymethyl-2-furaldehyde C6H6O3126 15.47 0.6
4-Methyl catechol C7H8O2124 17.21 1.5 2.0
Syringol C8H10O3154 18.69 0.5
Levoglucosan C6H10O5162 22.80 5.4
Caeine C8H10N4O2194 30.36 22.3 5.2
Theobromine C7H8N4O2180 30.75 1.8 1.0
DOI: 10.14218/JERP.2017.00015 | Volume 2 Issue 3, August 2017 75
Adesanwo JK.. et al: Medicinal activities of Cola nitida seed J Explor Res Pharmacol
seed of Cola nitida; however, current ndings revealed there are
yet more compounds that remain to be identied. The present work
has identied more chemical compounds from Cola nitida. The
presence of a number of FAs revealed in this publication serves as
evidence of the potential varied biological applications of the seed.
Recent reports on the biotechnological/nanotechnological applica-
tions of products of Cola nitida have opened the door to a new di-
mension of research activities involving the genus, especially with
respect to waste management. The kola nut pod, which hitherto was
considered a waste product, is being converted to useful products
– turning waste into wealth. With the aid of well-planned quantita-
tive HPLC analysis of the methanol extract of Cola nitida, some of
the unknown compounds can be isolated and structural elucidation
carried out. The chemicals identied in the kola nut can be deriva-
tised by functional group inter-conversion to more potent and less
harmful compounds for both industrial and medicinal applications.
Conclusions
Previous research on Cola nitida have established the presence of
xanthine alkaloids, catechins, epicatechins, anthocyanidins and
its oligomers (proanthocyanidins). However, from the ESI-MS
experiments, we discovered there are more compounds yet to be
identied and structurally elucidated. In this report, the FAs and
sugars present in Cola nitida have been identied using the GC-
MS (FAME and TMS derivatives of the extracts). With analytical
Py-GC-MS experiment, the constituents of thermal decomposi-
tion of Cola nitida were also identied and compared with those
of extracted biomass. In the antimicrobial assay, CH3OH extract
and solvent fractions showed no activity against Shigella Spp. but
were effective against most of the tested microbes. The N-hexane
fraction was ineffective in both antimicrobial and antioxidant as-
says. Caffeine was ineffective in antioxidant assay, but exhibited
appreciable activity against Bacillus cereus, Escherichia coli and
Aspergillus niger. The butanol fraction displayed higher antioxi-
dant activity than the CH3OH extract, which might be due to the
higher concentration of phenolic compounds in the butanol frac-
tion.
Acknowledgments
The corresponding author wishes to acknowledge the Tertiary
Education Trust Fund (TETFund) Conference grant from Obafemi
Awolowo University to present part of this report at the 4th Interna-
tional Pharma & Clinical Pharmacy Congress, November 07–09,
2016, Las Vegas, Nevada, USA.
Compound Molecular
formula
Molecular
weight
Retenon
me (min)
Kola
nut, %
Extracted
nut, %
Palmic acid C16H32O2256 32.36 1.5 0.8
Linoleic acid C18H32O2280 35.44 1.0
Oleic acid C18H34O2282 35.62 0.9 0.5
Stearic acid C18H36O2284 36.01 0.3
C18:2 C18H32O2280 36.67 0.3
C19:2 C19H34O2294 37.16 0.4
Methyl-2,3-dicyano-3- [4-dimethylamino)phenyl]-2-propenoate C14H13N3O2255 43.57 0.6
Squalene C30H50 410 46.34 0.4
Cholestadiene C27H44 368 47.28 0.1
Unknown steroid 344 47.50 0.1
Sgmastan-3,5-diene C29H48 396 49.89 0.3
Nortrachelogenin C20H22O7374 50.77 0.2
6,9,10-Trimethoxy-12H-[1]benzopino[2,3,4-ij]isoquinoline (oxocularine) C19H17NO4323 51.17 0.4
6,16 Dimethylpregna-1,4,6-triene-3,20-dione C23H30O2338 51.58 0.3
Sgmasterol C29H48O412 51.94 0.2
3,4,5,6-Tetrahydro alstonine C21H24N2O3352 52.26 0.4
Sitosterol C29H50O414 52.67 0.5
Table 4. Py-GC-MS of unextracted and extracted powdered Cola nida seed - (connued)
Compound 1. 1,3,7-trimethyl-1 H-purine-2,6(3H,7H)-dione (Caeine). Compound 2. n-Hexadecanoic acid.
DOI: 10.14218/JERP.2017.00015 | Volume 2 Issue 3, August 201776
Adesanwo JK.. et al: Medicinal activities of Cola nitida seed
J Explor Res Pharmacol
Conict of interest
The authors have no conict of interests related to this publication.
Author contributions
Originator of the research idea and supervising the work (JKA),
involving in isolation and antimicrobial experiments (SBO), as-
sisting in the antimicrobial study (DAA), involving in the chemi-
cal/instrumental analyses (AGM).
Supporting Information
Supplementary material for this article is available at https://doi.
org/10.14218/JERP.2017.00015.
Table S1. In vitro antimicrobial activity of the extracts and com-
pound 1 (caffeine) against selected microbes.
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... As a result, varying amounts of proximate composition, minerals, and amino acids have been reported in Cola nitida (Mbembo et al., 2022). One of the most well-known major components of Cola nitida is caffeine, a natural stimulant that can enhance alertness and cognitive function (Adesanwo et al., 2017;Okoli et al., 2012). The occurrence of caffeine in the seed of Cola nitida could potentially contribute to its utilization in the production of carbonated beverages (Asogwa et al., 2006;Jayeola, 2001;Ogutuga, 1975). ...
... The occurrence of caffeine in the seed of Cola nitida could potentially contribute to its utilization in the production of carbonated beverages (Asogwa et al., 2006;Jayeola, 2001;Ogutuga, 1975). In addition to caffeine, Cola nitida contains other alkaloids such as theobromine and theophylline, which contribute to its stimulating properties (Adesanwo et al., 2017;Ekalu & Habila, 2020;Ogutuga, 1975). Alkaloids have gained recognition for their therapeutic properties, notably their effectiveness as anesthetics, cardioprotective agents, and anti-inflammatory substances (Heinrich et al., 2021). ...
... Cola nitida contains various bioactive compounds that may have immunomodulatory properties. Flavonoids and phenolics found in Cola nitida have been shown to exhibit antioxidant and anti-inflammatory effects (Adedayo et al., 2019;Adesanwo et al., 2017;Oboh et al., 2014), which can help support immune system function. They can scavenge free radicals and reduce oxidative stress, which is implicated in the development of various diseases. ...
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Cola nitida, also known as Kola nut, is a tropical plant native to West Africa and has a rich history of traditional medicinal use. In this narrative review, we aim to provide an overview of the protective effects of Cola nitida in various health and disease states. Cola nitida has been traditionally used for its medicinal properties, and its bioactive compounds include caffeine, alkaloids, tannins, flavonoids, and phenolics. These compounds contribute to its potential therapeutic effects. Here, we examine the potential benefits of Cola nitida in several areas of health, discussing its role in cognitive function, cardiovascular health, immune system function, gastrointestinal health, and metabolic and endocrine health. Relevant original articles available from PubMed, African Journals Online (AJOL), SCOPUS, and Google Scholar were retrieved using the keywords “cola” AND “nitida” without date restriction until July 17, 2023. Evidence suggests that Cola nitida may have positive effects on health, with indications of adverse effects only from its chronic usage. However, more research is needed to establish its efficacy and safety. Cola nitida holds promise as a natural remedy for various health conditions. Understanding the benefits and limitations of Cola nitida will contribute to its effective utilization in health and disease management.
... Previous research has demonstrated the health benefits of Cola nitida [11]. It may have anticarcinogenic, antioxidant, antibacterial, and antidiabetic effects in addition to its capacity to prevent pituitary cells from releasing luteinizing hormones (LH), which may indicate that it has a function in reproductive regulation [12]. This study aims to explore the possible health advantages of the phytochemicals found in Cola nitida, specifically their potential as a cancer treatment intervention. ...
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Cancer continues to be the leading cause of mortality worldwide, despite significant attempts to identify new risk factors, create earlier diagnostic indicators, and investigate alternative therapy options. This rising burden of cancer worldwide results in the advancement of innovative methods of cancer prevention and treatment leading to the developed idea of cancer chemoprevention, with an emphasis on employing natural substances included in the diet to inhibit tumor development. Newer techniques like immunotherapy and gene therapy are used in addition to conventional cancer therapies including radiation, chemotherapy, and surgery. Nonetheless, the potential of natural substances needs to be exploited. Natural products which can be proteins, carbohydrates, or nucleic acids in their forms or conjugated have tendencies to trigger innate and adaptive immunity, infusion reactions, inflammatory responses, hypersensitivity reactions, and other immunological responses. Numerous bioactive substances found in Cola nitida, such as flavonoids, catechin, tannins, alkaloids, and phenolics, have been shown to have promising medicinal qualities, such as the ability to prevent cancer, function as antioxidants, and stop the proliferation of cancer cells using some of the immunological mechanism. This study explores the molecular routes by which these bioactive chemicals cause apoptosis, inhibit angiogenesis, and modify signal transduction pathways to achieve their anticancer effects. The study concludes by highlighting the tremendous multimodal therapeutic potential of Cola nitida in the creation of safer and more effective cancer treatments.
... Kolanut is dark brown in color when bitten, acidic taste when fresh and bitter when dry, with an increase in nutmeg and aromatic aroma 23 . Kolanut contains different fractions like catechins, procyanidins, sugar, sterols, fatty acids, alkaloids, kolanin, theobromine and caffeine in large quantity 24 . Kolanut has numerous uses, apart from being a social snack, it has been documented to reduce labor pain, swellings, and accelerate the healing of fresh wounds 25 . ...
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Kolanut contains caffeine and it is widely consumed in various social contexts in Nigeria and other Sub-Saharan African countries. While some studies have suggested that kolanut is consumed by pregnant women, there is a dearth of information on the prevalence, consumption pattern and reasons for kolanut consumption among this group. This study investigated kolanut use among pregnant women in Ibadan, Oyo State, Nigeria. A cross-sectional study involving 478 consenting pregnant women in all trimesters of pregnancy was conducted. Semi-structured questionnaires were used to collect data. Associations between kolanut use and respondent characteristics were investigated using the chi-square test and logistic regression analysis. The mean age of the women was 28.7 ± 6.3 years. One hundred and sixty-two (33.9%) of women reported kolanut use during pregnancy, 140 (29.3%) in the current pregnancy. Fifty-five (39.3%) pregnant women reported frequent use and 46 (32.9%) used it in high quantities. Significant associations were found between current kolanut use and Hausa respondents (p = 0.014), educational level; secondary (p = 0.032), tertiary (p = 0.006), TBA (p = 0.005). The majority (93.7%) used kolanut to prevent spitting, nausea, and vomiting. This study showed that kolanut use is quite common among pregnant women and frequently used in large quantities.
... The genus Cola has long been involved in Ayurvedic preparations which is based on the idea of herbal treatment and other natural therapies to treat various ailments and disorders. The genus has received the attention of pharmaceutical industries due to the presence of bioactive molecules like Hydroxy Citric Acid (HCA), oleic acid, flavonoids and theobromine which has immense remedial qualities 22,23 .The nuts are also rich in palmitic and oleic acids which are known to OPEN maintain good skin; consequently they are used in the production of cosmetic products such as soaps 24 , while the red coloured ones are regarded as a rich source of red pigments in the plant kingdom 25,26 . The pigments that give fresh foods their vibrant hues of red, green, purple, yellow and orange do more than just make a pretty meal, they contain powerful antioxidant properties that make a profound effect on the total health of consumers. ...
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Sexual incompatibility among kola genotypes accounted for over 50% yield loss. Compatible and high yielding varieties are in demand to develop commercial orchards. The objective of this study was to assess self-compatibility and cross-compatibility of kola (C. nitida) genotypes within self, single and double hybrid crosses and to determine heterosis pattern in the resulting hybrids for sexual compatibility and key nut yield and quality traits. Crosses among kola genotypes from three field gene banks (JX1, GX1, MX2) and one advanced germplasm (Bunso progeny) in Ghana were evaluated along their parents for sexual compatibility, nut yield and nut quality. Data were collected on pod set, pseudo-pod set, pod weight, number of nuts per pod, nut weight, brix, potential alcohol and nut firmness. Significant (P < 0.001) differential pod set was observed within Bunso progeny, JX1, GX1 and MX2 crosses; while pseudo-pod set differed only within JX1 and MX2 crosses (P < 0.001). Very large prevalence of mid-parent, heterobeltiosis, and economic heterosis was observed for sexual compatibility, outturn and brix for the single and double hybrid crosses. Heterosis was prominent among the double hybrid crosses as compared to the single hybrid crosses suggesting that recurrent selection of compatible varieties from advanced generations could result in genetic gain in kola improvement. The top five crosses with best heterosis for sexual compatibility and an appreciable positive heterosis for outturn and brix were B1/11 × B1/71 × B1/157 × B1/149, B1/11 × B1/71 × B1/296 × B1/177, GX1/46 × GX1/33 × B1/212 × B1/236, JX1/90 × JX1/51 and JX1/51 × JX1/36. These materials could serve as sources of beneficial alleles for improving Ghanaian kola hybrids and populations for yield and sexual compatibility.
... The efficacy of the plants mentioned in this study requires the mon- [40] who demonstrated the good antibacterial activity of the combination of extracts of C. nitida and fluoroquinolones on E. coli. The antibacterial activities shown by the species of C. nitida are also consistent with previous antimicrobial work [41,42,43] where crude extracts of C. nitida have been shown to be important inhibitors against the growth of certain bacteria (Escherichia coli, Staphylococcus aureus and Shigella dysenteriae). The water and water-ethanol extracts of C. nitida showed the greatest inhibitory activity against Enterococcus faecalis 103907 CIP compared to the water-acetone extract. ...
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The purpose of this work was to develop ethnopharmacological and biological studies of plants used in traditional medicine for the treatment of diarrheal diseases. This study results in scientific data that validates the uses of these plants in traditional medicine. Firstly, ethnopharmacological surveys were conducted with a few traditional healers from the provinces of Estuaire, Haut Ogooué and Woleu-Ntem in Gabon. Next, ethnobotanical data such as percentage of families, species, routes of administration, methods of preparation, parts used and number of plant names were analyzed and summarized. Finally, the antibacterial activities of some plants have been evaluated by diffusion and microdilution methods. Thirty-four (34) traditional healers were interviewed. A total of 90 plant species were identified during this study. They belong to 44 families, the most represented were Leguminoseae (13.33%), Apocynaceae (7.78%), Annonaceae (5.55%), Euphorbiaceae (4.44%) and Anacardiaceae (4.44%). Trees were used more (44.44%) than shrubs (32.22%), herbaceous plants (16.67%) and lianas (6.67%). The drug administration was mainly oral (84.62%) and by the anal route. Decoction and maceration were the two most used methods of preparation. Among identified plants, twenty-seven (27) plant extracts were subjected to microbiological analyzes. Plant extracts tested were active on Gram-negative and Gram-positive bacteria. Cola nitida extracts gave the best antibacterial activity against Enterococcus faecalis 103907 CIP. This study identified 90 antidiarrheal plant species and clearly shows the antimicrobial potential of several medicinal species.
... The medicinal properties of C. nitida are mainly grounded on chemical constituents of the plant roots and seeds. The plant contains several biochemical compounds including caffeine, theobromine, and theophylline which account for their medicinal properties (Adesanwo et al., 2017). The plant contains trace minerals such as calcium, iron, magnesium, manganese, phosphorus, potassium, sodium, and zinc some of which play significant roles as micro-and macronutrients essential for metabolic activities, growth, and development. ...
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The rural communities of the sub‐Sahara regions in Africa are rich in diverse indigenous culinary knowledge and foods, food crops, and condiments such as roots/tubers, cereal, legumes/pulses, locust beans, and green leafy vegetables. These food crops are rich in micronutrients and phytochemicals, which have the potentials to address hidden hunger as well as promote health when consumed. Some examples of these are fermented foods such as ogi and plants such as Vernonia amygdalina (bitter leaf), Zingiber officinales (garlic), Hibiscus sabdariffa (Roselle), and condiments. Food crops from West Africa contain numerous bioactive substances such as saponins, alkaloids, tannins, phenolics, flavonoids, and monoterpenoid chemicals among others. These bioresources have proven biological and pharmacological activities due to diverse mechanisms of action such as immunomodulatory, anti‐inflammatory, antipyretic, and antioxidant activities which made them suitable as candidates for nutraceuticals and pharma foods. This review seeks to explore the different processes such as fermentation applied during food preparation and food crops of West‐African origin with health‐promoting benefits. The different bioactive compounds present in such food or food crops are discussed extensively as well as the diverse application, especially regarding respiratory diseases. Practical applications The plants and herbs summarized here are more easily accessible and affordable by therapists and others having a passion for promising medicinal properties of African‐origin plants.The mechanisms and unique metabolic potentials of African food crops discussed in this article will promote their applicability as a template molecule for novel drug discoveries in treatment strategies for emerging diseases. This compilation of antiviral plants will help clinicians and researchers bring new preventive strategies in combating COVID‐19 like viral diseases, ultimately saving millions of affected people.
... The yield of this extraction was 12.6%. Adesanwo and al. [15] had found a higher yield (17.6 %) using methanol as solvent. With dichloromethane, they obtained a yield of 0.77%. ...
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Background: Native to West Africa, Cola nitida is a tropical tree of about 8-12 in height that grows in lowland rainforest. This plant is best known in Africa for its seeds, used in phytotherapy but for their socio-cultural importance. Aims/Objective: This study investigated the antioxidant activity of condensed tannins of Cola nitida seeds by carrying out two antioxidant tests (DPPH and FRAP). Methods: From a hydro-ethanolic extract of Cola nitida seeds, two samples were made. One treated with casein (EC) and another one without treatment (EWC). The researsh of condensed tannins were carried out by precipitation with Stiasny reagent. The total polyphenol and tannin contents were evaluated by the Folin-Denis method and the antioxidant power by DPPH and FRAP tests. Results: Extract without treatment (EWC) showed more antioxidant activity than the extract treated with casein (EC). Thus, the IC50 of EWC which contains condensed tannins was 5.54±0.005 µg/ml, while that of EC (without condensed tannins) reached 61.92±0.165 µg/ml. Conclusion: Cola nitida seeds are rich in condensed tannins that play an important role in the antioxidant activity.
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Since the beginning of the industrial revolution, the uncontrolled discharge of wastewater from textile and dyeing industries into water bodies is environmentally challenging to humans and the ecosystem. Using Cola nitida (CN) leaves extract, Taguchi optimization method was successfully applied to achieve the green synthesis of iron oxide nanoparticles (CN–Fe2O3NPs). Textural properties, nature of surface functional groups, crystalline structure and surface morphology of CN–Fe2O3NPs were studied. The performance of CN–Fe2O3NPs was tested for methylene blue (MB) and methyl orange (MO) removal from textile wastewater. The optimal surface area of 125.31 m2/g was achieved for the CN–Fe2O3NPs using the CN extract volume (10 mL), precursor concentration (2 M), contact time (30 min) and calcination temperature (600 °C). Overall, calcination temperature had the highest effect than other synthesis parameters. The characterization revealed the presence of crystalline hematite with Fe–O, Fe–O–Fe functional groups and a regular-shaped porous material. Furthermore, the adsorption capacity of 530.406 and 527.835 mg/g was obtained for MB and MO within 60 min using a nanoadsorbent dosage (25 mg/L), initial dye concentration (100 mg/L) and temperature (50 °C). The reusability and stability of the CN–Fe2O3NPs revealed successful reuse after six cycles without any damage to the structure as was corroborated by the fourier transform infrared (FTIR) spectroscopy. The experimental data were suitably fitted by the Langmuir isotherm and pseudo-second-order kinetic models denoting a chemisorption and monolayer nature of the adsorption process. The thermodynamic parameters indicate an endothermic and spontaneous adsorption process. Hence, the high separation effectiveness against the dye molecules with good performance and stability indicate the great potential for the green synthesized CN–Fe2O3NPs in water purification.
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This study investigates the influence of different concentrations of AgNPs biologically synthesized using pod extract of Cola nitida on antioxidant activity, phenolic contents, flavonoid contents and compositions of Amaranthus caudatus L. AgNPs of 25, 50, 75, 100 and 150 ppm were utilized in growing A. caudatus while water was used as control. Delayed germination for two days was observed for A. caudatus grown with 150 ppm of AgNPs, while others showed no difference. There were 43.3, 38.7, 26.7 and 6.48% improvements in the 2,2-diphenyl-1-picrylhydrazyl (DPPH) antioxidant activity of A. caudatus grown with 25, 50, 75 and 100 ppm of AgNPs, respectively, compared to control. Antioxidant activity of A. caudatus grown with AgNPs reduced with increase in the concentrations of AgNPs. A. caudatus grown with 50 ppm of AgNPs was the most potent with the least IC50 of 0.67 mg/ml. Significant improvements obtained for phenolic and flavonoid contents grown with AgNPs were concentration dependent. Enhancements of 21.9, 68.19, and 1.98% in phenolic contents were achieved in treatments with 25, 50 and 75 ppm AgNPs, respectively, while 32.58, 35.80, and 7.20% improvement in flavonoids were obtained for 25, 50 and 100 ppm treatments, respectively. Kaempferol and quercetin were the most abundant flavonoids in A. caudatus treated with 50 ppm of AgNPs, showing the highest flavonoid composition. This further confirms A. caudatus grown with 50 ppm of AgNPs as the most potent. This study has shown that concentration-dependent AgNPs can be used to boost antioxidant activity and phytochemical contents of vegetables.
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The present study examines the hydrogen peroxide scavenging, anticoagulant and thrombolytic activities of silver nanoparticles (AgNPs) that were biosynthesized using extracts obtained from spider cobweb (CB), pod (KP), seed (KS) and seed shell (KSS) of kolanut (Cola nitida). The nearly spherical shaped AgNPs, with surface plasmon resonance of 431.5–457.5 nm, were polydispersed having sizes of 3–50, 12–80, 8–50, and 5–40 nm for CB, KP, KS and KSS-AgNPs respectively. Hydrogen peroxide scavenging activities of 77–99.8% were obtained using 1–20 µg/ml of AgNPs. The particles prevented the coagulation of blood, and also showed thrombolytic activities of 55.76–89.83%, with KSS-AgNPs having the highest activity. Microscopic examination of the lyzed blood clot supported the thrombolytic activities. On the other hand, silver nitrate solution showed negligible activity of 1.92%, while thrombolysis of 7.55, 8.70, 8.93 and 30.19% were obtained for the extracts of KSS, CB, KS and KP respectively. The results herein presented showed potential biomedical applications of the biosynthesized AgNPs to scavenge free radicals and for the management of blood coagulation disorders and thrombotic diseases.
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This study investigated use of leaf, seed, seed shell and pod extracts of Cola nitida for the green synthesis of silver-alloy nanoparticles (Ag–AuNPs). The Ag–AuNPs formed were dark brown with maxima absorbance in the range of 497–531 nm. FTIR peaks at 3290–3396 and 1635–1647 cm−1 showed that proteins were the capping and stabilization molecules for the synthesis of Ag–AuNPs. While leaf, seed and seed shell extract-mediated Ag–AuNPs had near spherical morphology, anisotropic structures of sphere, rod, hexagon and triangle were formed by pod extract. The polydispersed particles were 17–91 nm in size, with crystalline characteristics and prominent presence of Ag and Au in the EDX spectra. Ag–AuNPs inhibited growth of Aspergillus flavus, A. fumigatus and A. niger by 69.51–100 %. Exposure of Anopheles mosquito larvae to Ag–AuNPs resulted in 80–100 % mortality in 24 h. Catalytic degradation of >90 and >60 % were obtained for malachite green and methylene blue respectively after 24 h. The particles displayed potent blood anticoagulant and thrombolytic activities, indicative of their potentials in the management blood coagulation disorders. This study showed that C. nitida can be used for green synthesis of Ag–AuNPs, which is the first report of its kind.
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This work reports the biogenic synthesis of silver nanoparticles (AgNPs) using pod extract of Cola nitida, evaluation of antibacterial activities, antioxidant activities and application as antimicrobial additive in paint. The AgNPs were characterized through UV-Vis spectroscopy, Fourier-Transform infrared spectroscopy (FTIR), and Transmission electron microscopy (TEM). The AgNPs was dark brown with maximum absorbance occurring at 431.5 nm. The FTIR spectrum showed strong peaks at 3336.85, 2073.48, and 1639.49 cm-1 indicating that proteins were the capping and stabilization molecules in the synthesis of AgNPs. The AgNPs were spherical with sizes ranging from 12-80 nm. The energy dispersive x-ray analysis showed presence of silver as a prominent metal, while the selected area electron diffraction pattern conformed to the face-centred cubic phase and crystalline nature of AgNPs. At various concentrations of 50-150 µg/ml, the AgNPs showed strong inhibition of growth of multidrug resistant strains of Klebsiella granulomatis, Pseudomonas aeruginosa, and Escherichia coli. In addition, at 5 µg/ml, the AgNPs totally inhibited growth of Staphylococcus aureus, E. coli, P. aeruginosa, Aspergillus niger, A. flavus and A. fumigatus in paint-AgNPs admixture. The AgNPs yielded potent antioxidant activities with IC50 of 43.98 µg/ml against 2, 2-diphenyl-1- picrylhydrazyl, and ferric ion reduction of 13.62-49.96 % at concentrations of 20-100 µg/ml. This study has demonstrated the biogenic synthesis of AgNPs with potent antimicrobial and antioxidant activities with potential biomedical and industrial applications. To the best of our knowledge, this work is the first reference to the use of pod extract of C. nitida for the green synthesis of nanoparticles
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IIn this study, seed and seed shell extracts of Cola nitida were investigated for the biogenic synthesis of silver nanoparticles (AgNPs) under ambient condition. The biosynthesized AgNPs were characterized through visual development of colour, UV-Vis using UV-Vis spectroscopy, Fourier-Transform infrared spectroscopy (FTIR), and Transmission electron microscopy (TEM). The antibacterial activities of the AgNPs were determined using some multi-drug resistant clinical isolates. The biosynthesized AgNPs depicted brown and yellowish orange colour using seed and seed extract respectively with maximum absorbance readings at 457.5 and 454.5 nm. The FTIR spectrum showed strong peaks at 3292.49, 2086.98, 1631.78 cm-1 for seed extract-mediated AgNPs, while peaks of 3302.13, 2086.05 and 1633.71 cm-1 were obtained for seed shell extract-mediated AgNPs, all indicating that proteins were the capping and stabilization molecules in the biogenic synthesis of AgNPs. The AgNPs were spherical in shape with sizes ranging from 8-50 and 5-40 nm for seed and seed shell-mediated AgNPs respectively. The energy dispersive x-ray (EDX) analysis showed the presence of silver as a prominent metal, while the selected area electron diffraction (SAED) pattern conformed to the face-centred cubic phase and crystalline nature of AgNPs. At various concentrations ranging from 50-150 µg/ml, the AgNPs inhibited growth of multi-drug resistant strains of Klebsiella granulomatis, Pseudomonas aeruginosa, and Escherichia coli to the tune of 10-32 mm. Comparatively, seed shell extract-mediated AgNPs had better activities with minimum inhibitory concentration (MIC) of 50 µg/ml against all the tested isolates, while the MIC of seed extract-mediated AgNPs were obtained as 50, 80, and 120 µg/ml against E. coli, P. aeruginosa (wound), and P. aeruginosa (burn) respectively. This study has demonstrated the feasibility of eco-friendly biogenic synthesis of AgNPs using seed and seed shell extracts of C. nitida, and the report to the best of our knowledge is the first reference to extracts of C. nitida for the green synthesis of AgNPs.
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This paper reviews the medicinal values of Kolanut in Nigeria with a view of identifying the most common species in the country and discussing the problem and Prospects of Kolanut trees. Some of the values of kolanut discussed include traditional value, nutritional value, economic/industrial value and the medicinal value which is the focus of this paper. The paper recommends that retraining efforts need be focused on the forestry extension to ensure that indigenous fruit trees like Kolanut become part of the basket of livelihood options supported by extension agents.
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This study was designed to take an inventory of medicinal plants, recipes and methods commonly used traditionally to treat some cardiovascular and inflammatory diseases in five local government areas in Ogbomoso, Oyo State, Nigeria. First-hand field survey through semi-structured questionnaire was employed in the five month study. A total of 101 plant species (medicinal plants (80.90%), spices (17.5%) and vegetables (1.53%)) belonging to 51 different families were mentioned for the treatment of various types of cardiovascular and inflammatory diseases. The survey revealed that 51.5% of the plants mentioned are used for the management of inflammatory diseases, 34.7% for the treatment of cardiovascular diseases and 11.9% of the plants are used for the treatment of both diseases. Euphorbiaceae (7.9%) are the most frequently used families of plants for the treatment of the various types of diseases mentioned, followed by Caesalpiaceae, (4.9%), Apocynoceae (4.9%) and Poaceae (4.9%). Fifty-nine recipes are usually prepared for the treatment of the six types of inflammatory diseases while twenty- three recipes are reportedly used for the treatment of the four types of cardiovascular diseases mentioned in this study. The recipes covered in the survey were mostly prepared from leaves (37.6%) and roots (23.8%) decoction or infusions. Medications are mostly administered orally with few numbers of the recipes showing side effect. The study has documented indigenous plants in Ogbomoso as a potential source for the development of new drugs for the treatment of cardiovascular and inflammatory diseases. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.