NIGERIAN AGRICULTURAL JOURNAL

ISSN: 0300-368X
Volume 55 Number 1, April 2024 Pg. 96-105
Available online at: [Link]
[Link]
Creative Commons User License CC:BY

Nutritional and Chemical Components of Ethanol Extract of Hunteria Umbellata Seeds

1
Anani, E. E., *2Shalem, S., 1Akpan, U. C. and 1David-Okwu, E.
1
Department of Biochemistry, Faculty of Basic Medical Sciences, University of Calabar
P.M.B. 1115, Calabar, Nigeria
2
Department of Nutrition and Dietetics, College of Food Technology and Human Ecology,
Joseph Sarwuan Tarka University, P.M.B. 2373, Makurdi, Benue State, Nigeria.
*Corresponding author's email: [Link]@[Link]
ORCID: [Link]

Abstract
Hunteria umbellata, a tropical rainforest tree commonly found in sub-Saharan Africa, is known for its medicinal
properties, particularly in treating diabetes, obesity, and anaemia. This study, a beacon of hope, investigated the
nutritional and chemical composition of the ethanol extract of Hunteria umbellata seeds. The research, conducted
with unwavering dedication, focused on proximate, phytochemical, vitamin, and mineral analyses of the seed
extract, all of which were analysed using standard analytical procedures. The proximate analysis revealed that
carbohydrates were the most abundant component (56.73±0.24%), followed by moisture (16.20±0.16%) and
crude protein (13.92±0.01%). Fibre content was 8.03±0.41%, with a calorific value of 1277.43±3.18 KJ/100g.
Ash and lipid content were the lowest at 2.53±0.06% and 2.59±0.02%, respectively. Qualitative phytochemical
analysis identified the presence of alkaloids, flavonoids, reducing sugars, saponins, tannins, phytates and
oxalates. Quantitative analysis highlighted high concentrations of flavonoids (383.50±0.24 mg/100g) and
tannins (14.02±0.01 mg/100g). GC-MS identified 20 compounds, with phenol 2,4-bis (1,1-dimethyl ethyl)
predominant (37.72%). Vitamin analysis indicated substantial amounts of vitamin K (135.00±0.14 IU/100g),
vitamin A (21.95±0.01 IU/100g), and vitamin D (8.30±0.14 IU/100g). The B complex vitamins included
thiamine-B1, folic acid-B9, biotin-B7, pyridoxine-B6, riboflavin-B2, and cobalamin-B12. Mineral analysis
showed significant amounts of zinc (7.08±0.11 mg/L), iron (3.51±0.02 mg/L), calcium (7.74±0.08 mg/L),
magnesium (4.22±0.21 mg/L), phosphorus (3.31±0.01 mg/L), potassium (9.37±0.22 mg/L), and sodium
(80.59±0.66 mg/L), with sodium being the highest. These findings, a ray of hope, provided valuable insights into
the nutritional and chemical properties of Hunteria umbellata seed extract, thereby supporting its potential health
benefits and therapeutic applications. Further studies are recommended to explore its therapeutic applications
and clinical efficacy, which are promising prospects for the future.
Keywords: Hunteria umbellata, ethanol extraction, proximate, phytochemicals, micronutrients

Introduction The seeds of H. umbellata, traditionally used in various

Medicinal plants globally are valuable sources of new forms of medicine, particularly in Nigeria, hold
drugs due to the rising demand for natural health significant potential for therapeutic applications.
products and herbal medications. Among several other Known as 'Osu' in Edo and 'Abeere' in Yoruba dialects,
medicinal plants in Africa, the different parts of the the plant's rich phytochemical and nutritional
Hunteria umbellata plant have been explored to cure composition is the key to its potential. Phytochemical
human diseases. H. umbellata is a member of the analysis of the seed extract has revealed the presence of
Apocynaceae plant family and is frequently available in various bioactive compounds, including tannins,
many sub-Saharan African countries, including Nigeria. saponins, steroids, and flavonoids, which have been
It is a small tree with dense, hard-textured leaves, dark linked to a range of biological activities, such as
brown stems, abundant flowers, and large fruits-bearing antimicrobial, antioxidant, and anti-inflammatory
seeds (Morakinyo et al., 2020; Udinyinwe & Aghedo, effects (Udinyinwe & Aghedo, 2022). These findings
2022; Ahajumobi et al., 2022). pique the audience's interest and instil hope in the
potential health benefits of Hunteria umbellata seed

------------------------------------------------------------------------------------------------------------------------------------------------------
Anani, Shalem, Akpan & David-Okwu
Nigerian Agricultural Journal Vol. 55, No. 1 | pg. 96
extract. Proximate Composition of Ethanol Extract of H.
umbellata Seeds
The proximate composition of the seed extract is also The proximate composition of ethanol extract of H.
noteworthy. The seeds contain significant amounts of umbellata seeds was analysed as follows: Protein
carbohydrates, proteins, and minerals such as calcium, content was quantified using the MicroKjeldahl method,
phosphorus, potassium, and zinc. These nutrients are encompassing sample digestion, distillation, and
essential for various physiological processes, including titration. Fat and oil content were determined following
energy production, bone health, and immune function the AOAC method (2010). Moisture content was
(Morakinyo et al., 2020; Udinyinwe & Aghedo, 2022; measured by indirect distillation, where a 2.0g sample
Ahajumobi et al., 2022). Furthermore, the seed extract was dried in an oven at 40°C until a constant weight was
has been found to contain various anti-nutrients like achieved and calculated using the equation: Moisture =
oxalate and phytate, which can affect nutrient (loss in weight)/(weight of the sample (g)) × 100. The
absorption and utilisation (Morakinyo et al., 2020; crude fibre was assessed through sequential treatments
Udinyinwe & Aghedo, 2022). with light petroleum, boiling dilute sulphuric acid,
boiling dilute sodium hydroxide, dilute hydrochloric
Given the potential therapeutic and nutritional benefits acid, alcohol, and ether, with the percentage calculated
of H. umbellata seed ethanol extracts, this study is a as Crude fibre (%) = (w1 - w2)/w1 × 100. Total ash
crucial step in understanding the nutritional and content was measured by igniting the sample in a muffle
chemical composition of the ethanol extract. furnace at 550-600°C, calculated using the equation:
Specifically, this research will focus on the seed extracts' Ash (%) = (weight of ash)/(weight of the sample (g)) ×
phytochemical, proximate, and micronutrient contents 100. Available carbohydrates were determined by
to better understand their biological activities. The need difference, with % NFE = 100 - (% ash + % crude fat + %
for further studies is evident, and the potential impact of crude fibre + % crude protein + % moisture).
this research on the field of pharmacology, botany, and
nutrition is significant, making the audience feel the Qualitative Phytochemical Analysis
importance of their work and the potential impact of The qualitative phytochemical analysis of the ethanol
their research. extract of H. umbellata seeds was conducted using
established methods (Harborne, 1973; Trease & Evans,
Materials and methods 1989; Sofowora, 1993; Deka & Kalita, 2012). To
Materials determine tannin content, 1.0 ml of the extract was
All the chemicals used in this study, including FeCl3 and diluted with distilled water (4:1) and treated with FeCl3,
disulfiram, were of analytical grade and sourced from where a blue or green colouration indicated tannins. For
May and Baker (England), Merck (Germany), Sigma- saponins, 5 ml of distilled water was added to 2.0 ml of
Aldrich (Germany), BDH (UK), and MN Kieselgel the extract and shaken vigorously with olive oil, and
GmbH (Germany). The remaining reagents used in emulsion formation confirmed saponins. Flavonoids
various experiments were purchased from Merck were detected by adding 2.0 ml of 5% NaOH to the
(Germany), Randox Laboratories (UK), Biovendor extract, resulting in a yellow colour. Alkaloid presence
(Czech Republic), and Teco Diagnostics (USA). was indicated by the formation of an orange colour upon
Glassware and equipment included Randox commercial adding Dragendorf reagent to 2.0 ml of the extract.
kits from Randox Laboratories Limited, UK, a Steroids were identified by adding 2 ml of acetic
spectrophotometer (722 N, China), and a haematology anhydride to 0.5 g of the extract, followed by sulfuric
analyser (BC-2300, Mindray Medical CO., China). H. acid, resulting in violet or blue-green colouration.
umbellata seeds were obtained from Urua Ekpat, Uyo, Phenols were confirmed by mixing the extract with
Akwa Ibom State, and subsequently authenticated by distilled water, warming it, and adding 2 ml of ferric
the Department of Plant and Ecological Studies, Faculty chloride solution, which produced a green or blue
of Biological Sciences, University of Calabar, Calabar, colour. Finally, cardiac glycosides were detected by
where they were assigned the voucher number adding 0.5 ml of the extract to 1 ml of glacial acetic acid
Bot/Herb/Ucc/095. with ferric chloride, followed by concentrated sulfuric
acid, and observing a reddish-brown colour at the
Methods junction and a bluish-green upper layer.
Ethanol extraction of the powdered seed of H.
umbellata Quantitative Phytochemical Analysis
Seed extraction was carried out according to the method The quantitative phytochemical analysis of ethanol
reported by Gahlot et al., (2018). Briefly, the seeds were extract of H. umbellata seeds involved several
washed, air-dried under shade, and then milled into determinations. Saponin content was assessed using the
powder using a manual blender. The powdered seeds method of Obadoni and Ochuko (2002). A 50 mg sample
were extracted with ethanol (using a Soxhlet apparatus). was combined with 100 cm³ of 20% aqueous ethanol
The solvent in the extract was evaporated using a rotary and heated at 55°C for 4 hours with continuous stirring.
evaporator and a water bath (50°C). The extract was The mixture was filtered and re-extracted with 200 ml of
weighed, and the per cent yield was calculated. 20% ethanol, and the combined extracts were
concentrated to 40 ml. This filtrate was transferred to a
separating funnel, mixed with diethyl ether, and the

------------------------------------------------------------------------------------------------------------------------------------------------------
Anani, Shalem, Akpan & David-Okwu
Nigerian Agricultural Journal Vol. 55, No. 1 | pg. 97
aqueous layer was retained. The process was repeated, 300°C at 10°C/min. One microliter of the prepared 1%
and 60 ml of n-butanol was added, followed by washing extract solution was injected in splitless mode. The
with 10 ml of 5% aqueous sodium chloride. The solution relative quantities of chemical compounds in each
was heated, evaporated, and dried to constant weight to extract were determined by calculating the percentage
determine the saponin content. Alkaloid content was area of each peak in the chromatogram. Bioactive
determined according to Harborne (1973). A 2.5 g compounds were identified by comparing the GC
sample was treated with 200 cm³ of 10% acetic acid in retention times with those of standards in the GC-MS
ethanol and left to stand for 4 hours. The extract was system's Replib and Mainlab data libraries (Oshiobugie
concentrated, precipitated with concentrated et al., 2019).
ammonium hydroxide, and filtered. The precipitate was
washed, dried, and weighed. Flavonoid content was Vitamin composition of ethanol extracts of Hunteria
evaluated using the Boham et al. (1974) method, where umbellata seeds
a 50 mg sample was extracted with 100 ml of 80% The amount of vitamins A, E, C, and B12 in the sample
aqueous methanol, filtered, evaporated, and weighed. was determined using the method described by
Total phenolic content was measured using the modified Achikanu et al., (2013) and AOAC (2010.) Vitamin B1
Folin-Ciocalteu method (Lawag et al., 2023), with and B3 were determined using the method described by
absorbance read at 765 nm and results expressed as Okwu and Ndu (2006), while Vitamin B2, K, and folate
tannic acid equivalents. Cardiac glycosides were were determined using the methods described by Okwu
quantified following Harborne (1973), involving and Josiah (2006). The method described by Oulai et al.,
filtration, addition of water, glacial acetic acid, FeCl3, (2014) was used to determine β-carotene.
and H SO , with absorbance measured at 410 nm.
Steroid content was determined using the Madhu et al. Mineral composition of ethanol extract of Hunteria
(2016) method, involving treatment with sulphuric acid, umbellata seed
iron (III) chloride, and potassium hexacyanoferrate Phosphorus was quantified using the method outlined by
(III), heated and measured at 780 nm. Terpene content Oshodi, 1999. Approximately 10mg of the sample was
was assessed using the Mboso et al. (2013) method, extracted overnight in 250ml of concentrated nitric acid
where a 1 g sample was extracted with petroleum ether and then per choric acid on a hot plate. Distilled water
and measured at 420 nm. Lastly, tannin content was (20ml) was added to the sample and boiled until white
measured by weighing 50 mg of the sample, extracting it fumes appeared. This was followed by adding 5ml of
with distilled water, and reacting it with FeCl3 and K3Fe ammonia solution and further boiling until crystals
(CN)6, with absorbance read at 120 nm. formed. The crystals were dissolved using 20ml of
acidified water and 80ml of distilled water, followed by
Gas Chromatography-Mass Spectrometry (GC-MS) 20ml of mixed reagent for colour development. The
analysis of ethanol seed extract of Hunteria umbellate mixed reagent consisted of 250ml sulphuric acid-
for chemical compounds present antimony, 50ml distilled water and 2gl ascorbic acid.
For the Gas Chromatography-Mass Spectrometry (GC- The sample was left for 15 minutes for the colour to
MS) analysis of the ethanol seed extract of Hunteria develop. The quantity of phosphorus was then
umbellata, 1 g of the sample was weighed and determined using a spectrophotometer (Cecil Elegant
transferred to a test tube, followed by adding 15 mL of Technology, UK). Other minerals (calcium, iron,
ethanol. The mixture was incubated in a water bath at magnesium, manganese, sodium, potassium and zinc)
60°C for 60 minutes. Post incubation, the reaction were quantified using a thermo x series 2 inductively
mixture was transferred to a separating funnel. The test coupled plasma mass spectrophotometer (ICP MS)
tube was sequentially rinsed with 20 mL of ethanol, 10 (Thermo Scientific, USA). Samples weighing
mL of cold water, 10 mL of hot water, and 3 mL of approximately 80mg were digested in nitric acid for 1hr
hexane, all transferred to the funnel. These extracts were in a mass Xpress microwave (CEM Corporation, USA)
combined and subjected to three washes with 10 mL of and then diluted to 10ml with distilled water ready for
10% v/v ethanol aqueous solution. The solution was qualification.
dried using anhydrous sodium sulphate, and the solvent
was evaporated. The dried sample was reconstituted in Results
1000 μL of ethyl acetate, and 200 μL of this solution was Proximate composition of ethanol extract of H.
transferred to a vial for GC-MS analysis (Oshiobugie et umbellata seed
al., 2019). Table 1 presents the proximate composition of ethanol
extract of H. umbellata seeds. Carbohydrates exhibited
Quantification of phytochemicals was carried out using the highest percentage at 56.73±0.24 %, followed by
a BUCK M910 Gas Chromatograph equipped with an moisture (16.20±0.16 %) and crude protein (13.92±0.01
HP-5MS column (30 m length × 250 μm diameter × 0.25 %). The fibre content of H. umbellata seeds was
μm film thickness). Detection was performed using an recorded as 8.03±0.41 %, while the calorific value was
electron ionisation system at 70 eV. Helium gas determined to be 1277.43±3.18 KJ/100g. On the other
(99.995% purity) was used as the carrier gas at a 1 hand, the ash and lipid content of the ethanol extract of
mL/min flow rate. The GC temperature program was H. umbellata seeds showed the lowest values at
initiated at 50°C, increased to 150°C at a rate of 3°C/min 2.53±0.06 % and 2.59±0.02 %, respectively.
with a hold time of 10 minutes, and then ramped to

------------------------------------------------------------------------------------------------------------------------------------------------------
Anani, Shalem, Akpan & David-Okwu
Nigerian Agricultural Journal Vol. 55, No. 1 | pg. 98
Qualitative phytochemical analysis of ethanol sodium (80.59±0.66 mg/L). Among these minerals,
extract of H. umbellata seed sodium exhibits the highest concentration, with a
Table 2 presents the preliminary qualitative sodium-potassium ratio (Na: K) 8.60.
phytochemical analysis of the ethanol extract obtained
from H. umbellata seeds. The analysis revealed the Discussion
presence of alkaloids, flavonoids, reducing sugar, Medicinal plants are important in traditional medicine
saponins, tannins, phytates, oxalates and cyanogenic and are often used as home remedies (Welz et al., 2018).
glucosides. Notably, the extract exhibited a prominent The use of herbal medicinal products has increased in
presence of flavonoids and saponins. recent years due to their low side effects, ease of access
and acceptance, especially in developing countries
Quantitative phytochemical analysis of ethanol (Sofowora et al., 2013; Ozioma & Chinwe, 2019).
extract of H. umbellata seed Today, many people worldwide use medicinal plants
The result of selective quantitative phytochemical extensively to alleviate and treat diseases due to their
screening of ethanol extract of H. umbellata seeds is gentle nature and limited side effects (Mintah et al.,
presented in Table 3. Among the analysed compounds, 2019). These plants are rich in natural antioxidants such
the extract exhibited the highest concentration of as phenolic compounds, tocopherols, carotenoids and
flavonoids (383.50±0.24 mg/100g), saponins ascorbic acid (Lourenço et al., 2019). Research has
(150.27±0.11 mg/100g) and tannins (14.02±0.01 shown that consuming natural antioxidants can reduce
mg/100g). Relatively lower amounts of oxalates the risk of various diseases (Zhou et al., 2021).
(10.42±0.17 mg/100g), cyanide (6.21±0.04 mg/L), Generally, vitamins such as tocopherols, carotenoids
alkaloids (1.80±0.28 mg/100g), and phytates and ascorbic acid are active components that have a
(0.26±0.05 mg/100g) were found in the extract. protective effect (Okwu, 2005; Liu et al., 2018).

Gas Chromatography-Mass Spectrometry (GC-MS) Hunteria umbellata (K. Schum.), a member of the
analysis of ethanol Hunteria umbellata seed extract Apocynaceae family, is well-known in folklore for its
Table 4 shows the chemical composition of the ethanol traditional use in managing labour, pain and swellings,
seed extract of Hunteria umbellata, identified using Gas stomach ulcers, diabetes, obesity, and anaemia (Falodun
Chromatography-Mass Spectrometry (GC-MS). About et al., 2006; Adejuwon & Olufunmilayo, 2009). These
20 compounds were identified from the ethanol extract therapeutic properties have been attributed to various
of H. umbellata seeds. The main compounds identified bioactive compounds, vitamins, and minerals in the
based on the percentage contents were phenol, 2,4-bis plant's seeds. However, there is a lack of comprehensive
(1,1-dimethyl ethyl) (37.72%), 4-Amino-7- scientific information regarding the physicochemical,
diethylamino-chromen-2-one (10.75%), trans-13- phytochemical, and reversibility profile of the ethanol
Octadecenoic acid (9.47%), (Z)-9-Octadecenamide extract derived from Hunteria umbellata seeds.
(8.71%), cis-Vaccenic acid (6.13%), 1-(ethenyloxy)- Therefore, the main objective of this study was to
Octadecane (3.17). investigate the proximate, phytochemical composition
(both quantitative and qualitative), mineral and vitamin
Vitamin composition of ethanol extract of Hunteria of the ethanol extract obtained from H. umbellata seeds.
umbellata seeds According to the proximate composition result of H.
Table 5 presents the analysis of the vitamin content of umbellata seeds, it was found that they contain a high
the Hunteria umbellata seeds' ethanol extract. The result percentage of available carbohydrates (56.73%), which
demonstrates that the seeds possess substantial amounts was in agreement with the research carried out by
of vitamin K (135.00±0.14 IU/100g), vitamin A Morakinyo et al., (2020), while ash was present in the
(21.95±0.01 IU/100g), and vitamin D (8.30±0.14 lowest amount (2.53%). The ash content in the seed
IU/100g). However, vitamin E (1.25±0.07 mg/100g) extract (2.53%) was similar to the values obtained for
and beta-carotene (0.41±0.00 mg/100g) were present in Prunus persica seed (3.36%) (Ashraf et al., 2011),
smaller quantities. The vitamin B complex content of calabash seed (3.70%) (Abolaji et al., 2007), and
the seed under study is shown in Table 6. The detected calabash whole seed (4.0%) (Oyeleke et al., 2011). The
vitamins include thiamine-B1 (0.0446 mg/100g), folic ash content indicates the presence of inorganic elements
acid-B9 (0.3375 mg/100g), biotin-B7 (8.789 mg/100g), in the seeds. The ash content in H. umbellata seeds was a
pyridoxine-B6 (0.2025 mg/100g), riboflavin-B2 little above the recommended threshold of 1.5-2.5% for
(88.5138 mg/100g), and cobalamin-B12 (0.5757 animal feed, as suggested by Pomeranz and Cliffton
mg/100g). (2015), who stated that seeds with an ash content above
2.5% are unsuitable for animal consumption.
Mineral composition of ethanol extract of Hunteria
umbellata seeds Furthermore, the carbohydrate content of H. umbellata
The selected mineral composition of the ethanol extract seeds (56.73%) was higher than the values reported for
of Hunteria umbellata seeds is shown in Table 7. The Adansonia digitata (44.60%), baobab pulp seed
results reveal appreciable amounts of zinc (7.08±0.11 (44.6%), and Prunus persica (47.44%) (Oyeleke et al.,
mg/L), iron (3.51±0.02 mg/L), calcium (7.74±0.08 2012). Carbohydrates serve as a significant energy
mg/L), magnesium (4.22±0.21 mg/L), phosphorus source. The protein content of H. umbellata seeds
(3.31±0.01 mg/L), potassium (9.37±0.22 mg/L) and (13.92%) was similar to that reported for Cola millenii,

------------------------------------------------------------------------------------------------------------------------------------------------------
Anani, Shalem, Akpan & David-Okwu
Nigerian Agricultural Journal Vol. 55, No. 1 | pg. 99
Megaphrynium mascosterchyum, and Rauwolfia for their astringent properties and potential to treat
Victoria, which are 12.52%, 10.78%, and 8.65%, diarrhoea and other gastrointestinal disorders. However,
respectively. However, it was lower than the protein they can also inhibit the absorption of certain nutrients,
content of Ceasalpinia bonduc (19.67%), baobab pulp necessitating a balance in their consumption (Riaz et al.,
seed (19.05%), and Prunus persica (19.70%) (Ajayi et 2023). The presence of anti-nutrients, such as oxalates
al., 2015). Protein plays a crucial role in tissue repair (10.42±0.17 mg/100g), cyanogenic glycosides
within the body. The crude fibre content of the seeds was (6.21±0.04 mg/L), alkaloids (1.80±0.28 mg/100g), and
determined to be 8.03%, which was lower than the value phytates (0.26±0.05 mg/100g), although in relatively
reported for the dehulled seed of calabash (23.90%) lower amounts, is noteworthy. These anti-nutrients can
(Oyeleke et al., 2011) but close to the values reported for potentially interfere with the bioavailability and
Canna bidentata (12.68%) and baobab seed (15.6%). utilisation of certain nutrients and may pose toxicity
The high crude fibre level in H. umbellata seeds risks at higher levels. Their levels should be considered
indicated their potential to maintain proper digestion. It when evaluating the safety and potential applications of
may positively affect managing conditions such as the H. umbellata seed extract. Therefore, the
diabetes, cholesterol absorption, cardiovascular consumption of H. umbellata seeds should be
disease, colorectal cancer, and obesity (Ganong, 2003). moderated, and further studies are needed to understand
the safe dosage and potential side effects (Salisu et al.,
The moisture content of H. umbellata seeds (16.20%) 2024).
was higher than that reported for baobab pulp seed
(11.2%) (Oyeleke et al., 2013) and calabash whole seed The ethanol seed extracts of H. umbellata were
(9.2%) (Oyeleke et al., 2011). This suggested that the subjected to gas chromatography-mass spectrometry
seeds have good storage stability. The lipid content in H. (GC-MS) analysis, resulting in the identification of 19
umbellata seeds was reported as 2.59%, comparable to different compounds. The predominant compounds of
the values reported for Solanum dasyphyllum (2.65%) interest, based on their percentage contents, were
and Canna bidentata (3.25%). However, it was lower phenol, 2,4-bis(1,1-dimethylethyl) (37.72%), 4-Amino-
than the lipid content of Hydrocotyle aziata (8.15%) 7-diethylamino-chromen-2-one (10.75%), trans-13-
(Ajayi et al., 2015) and lower than that of baobab pulp Octadecenoic acid (9.47%), (Z)-9-Octadecenamide
seed (13.4%). Due to the low-fat content in H. umbellata (8.71%), cis-Vaccenic acid (6.13%), and 1-
seeds, they may have potential benefits in reducing (ethenyloxy)-Octadecane (3.17%). As a natural
obesity and maintaining healthy skin. compound, Phenol, 2,4-bis(1,1-dimethylethyl) has been
reported to possess various medicinal, food, and
The phytochemical analysis of the ethanol extract of agricultural functions. It exhibits antioxidant properties
Hunteria umbellata seeds has revealed a rich (Choi & Lee, 2009), anticancer activity (Pereira et al.,
composition of bioactive compounds consistent with 2009; Ren et al., 2019), antifungal properties (Zhou et
the plant's traditional use in herbal medicine. The al., 2011; Ren et al., 2019), antibacterial activity
qualitative analysis indicated a strong presence of (Abdullah et al., 2011; Nathar et al., 2018), and provides
flavonoids and saponins. The prominence of these protection against trimethyltin (TMT)-induced
compounds is significant, as flavonoids are known for cognitive dysfunction (Kim et al., 2017). Furthermore,
their antioxidant properties, which play a crucial role in 4-Amino-7-diethylamino-chromen-2-one has been
combating oxidative stress and related chronic diseases. found to exert diverse biochemical and physiological
On the other hand, Saponins have been recognised for effects. It displays anti-inflammatory, antioxidant, and
their cholesterol-lowering abilities and potential to anticancer activities and neuroprotective and
boost the immune system. Flavonoids, renowned for cardioprotective effects (Vidhu & Evans, 2015).
their antioxidant properties, could potentially contribute Additionally, it exhibits immunomodulatory and anti-
to preventing chronic diseases such as cancer and diabetic effects (Ansary & Taher, 2019). Moreover, (Z)-
cardiovascular diseases. (Fadahunsi et al., 2021). The 9-octadecenamide exhibits potent anti-inflammatory
high flavonoid content (383.50+0.24 mg/100g) in H. properties (Ano et al., 2015) and demonstrates analgesic
umbellata seeds suggests that the plant could be a potent and ulcerogenic effects on health (Hadi et al., 2016).
source of natural antioxidants1. This is further However, Cis-vaccenic acid, an omega-7 7-fatty acid,
supported by a study that screened and analysed the anti- contains antioxidant properties, which involve the
hyperlipidaemic potential of ethanolic extracts of H. neutralisation of free radicals and a reduction in
umbellata seeds, indicating that the plant's oxidative stress (Malek et al., 2009). Also, it displays
phytochemicals could be harnessed (Morakinyo et al., anti-inflammatory, anticancer, antidiabetic and anti-
2020). obesity properties (Zahid et al., 2017).

Saponins, another prominent compound (150.27±0.11) The findings of this research regarding the nutritional
in H. umbellata seeds, have been recognised for their composition of H. umbellata aligned with the results of
cholesterol-lowering and immune-boosting effects. The Morakinyo et al. (2020), indicating the presence of
presence of saponins in the extract aligns with the plant's essential minerals like calcium, iron, sodium,
reported use in traditional medicine for treating various phosphorus, magnesium, and zinc. These elements
ailments (Fadahunsi et al., 2021). Tannins, also found in found in the seeds of H. umbellata are beneficial for
significant amounts (14.02±0.01 mg/100g), are known disease prevention and control, maintaining acid-base

------------------------------------------------------------------------------------------------------------------------------------------------------
Anani, Shalem, Akpan & David-Okwu
Nigerian Agricultural Journal Vol. 55, No. 1 | pg. 100
balance, regulating osmotic pressure, facilitating nerve extract in experimental diabetes. Journal of
impulse conduction, aiding muscle contraction Ethnopharmacology. 10:1016
(especially the cardiac muscle), regulating bones and Ahajumobi, N. E., Oparaocha, T. E., Eteike, P., Felix, S.
teeth, and supporting cell membrane function (Murray O. (2022). Effect of water intake on constipation
et al., 1999). and bowel movement. Asian Journal of Medicine
and Health. 20(9):1-10.
In terms of vitamins, the analysis revealed that H. Ajayi, I. A., & Ojelere, O. O. (2015). Phytochemical
umbellata seeds contain high amounts of vitamins A analysis and mineral element composition of ten
(21.95 IU/100g), E (1.25 mg/100g), and K (135.00 medicinal plant seeds from South-West Nigeria. NY
IU/100g). These findings were consistent with the Science Journal, 6(9), 22-28.
results obtained by Morakinyo et al. (2020), which also Ano, Y., Ozawa, M., Kutsukake, T., Sugiyama, S.,
indicated the abundant presence of vitamins A, E, and K Uchida, K., Yoshida, A., & Nakayama, H. (2015).
in H. umbellata seeds. Vitamin E acts as an antioxidant Preventive effects of a fermented dairy product
that inhibits the progression of free radical reactions against Alzheimer's disease and identification of a
involved in various disease processes (Herrera & novel oleamide with enhanced microglial
Barbas, 2001). Vitamin A, a group of retinoid phagocytosis and anti-inflammatory activity. Plos
compounds with the biological activity of all-trans- One, 10(3), e0118512.
retinol, plays a crucial role in physiological functions Ansary, I., & Taher, A. (2019). One-Pot Synthesis of
such as vision, growth, reproduction, haematopoiesis, Coumarin Derivatives. In V. Rao, D. Mans, & L.
and immunity (Villamor & Fawzi, 2005). Notably, Rao (Eds.), Phytochemicals in Human Health (p.
vitamins A and E (a-tocopherol) have been recognised C h . 6 ) . I n t e c h O p e n .
as chemo-preventive agents against certain types of [Link]
cancer (Sirimanne, 1998). AOAC (2010) Official Methods of Analysis of
Association of Official Analytical Chemists. 18th
In this study, the analysis of the vitamin B complex in H. Edition, Washington, DC.
umbellata seeds revealed that Riboflavin-B2 and Ashraf, C. M., Iqbal, S., & Ahmed, D. (2011).
Biotin-B7 were the most abundant. The B vitamins Nutritional and Physicochemical Studies on Fruit
function as coenzymes in various enzymatic processes pulp, Seed and Shell of Indigenous Prunus persica.
essential for cellular physiological functions, including Journal of Medicinal Plants Research, 5(16), 3917-
vital functions within the brain and nervous system 3921.
(Hanna et al., 2022). Deficiencies in any of the B Boham B. A. & Kocipal-Abyazan R. (1974). Flavonoids
vitamins can adversely affect the mitochondrial and Condensed Tannins from Leaves of Hawaiian
metabolism of amino acids, glucose, and fatty acids vaccinicum vaticulatum and V. calycium. Pacific
through the citric acid cycle and electron transport chain Science, 48: 458-463.
(Kennedy, 2 016). Choi, Y., & Lee, J. (2009). Antioxidant and
antiproliferative properties of a tocotrienol-rich
Conclusion fraction from grape seeds. Food Chemistry, 14,
In this study, we analysed the ethanol extract of H. 1386.
umbellata seeds to understand their composition and Deka, M., & Kalita, J. C. (2012). Preliminary
potential therapeutic benefits. The seeds were rich in Phytochemical Analysis and Acute Oral Toxicity
carbohydrates, proteins, crude fibre, minerals, and Study of Mucuna pruriens Linn. in Albino Mice.
vitamins, making them nutritionally valuable. The International Research Journal of Pharmacy, 3(2),
presence of bioactive compounds such as alkaloids, 181–183. [Link]
flavonoids, glycosides, saponins, and phenols suggests Fadahunsi, O. S., Adegbola, P. I., Olorunnisola, O. S.,
potential therapeutic benefits. Gas chromatography- Subair, T. I., Adepoju, D. O., & Abijo, A. Z. (2021).
mass spectrometry analysis identified specific bioactive E t h n o - m e d i c i n a l , p h y t o c h e m i s t r y, a n d
compounds associated with medicinal properties. pharmacological importance of Hunteria umbellate
(K. Schum.) Hallier f. (Apocynaceae): a useful
References medicinal plant of sub-Saharan Africa. Clinical
Abdullah, A. S. H., Mirghani, M. E. S., & Jamal, P. P h y t o s c i e n c e , 7 ( 1 ) , 1 – 1 7 .
(2011). Antibacterial activity of Malaysian mango [Link]
kernel. African Journal of Biotechnology, 10, Falodun, A., Nworgu, Z. A., & Ikponmwonsa, M. O.
18739. (2006). Phytochemical components of Hunteria
Abolaji, O. A., Adebayo, A. H., & Odesanmi, O. S. umbellata (K. Schum) and its effect on isolated
(2007). Nutritional Qualities of Three Medicinal non-pregnant rat uterus in oestrus. Pakistan
Plant Parts (Xylopia aethiopica, Blighia sapida, and Journal of Pharmaceutical Sciences, 19(3),
Parinari polyandra) commonly used by Pregnant 256–258.
Women in the Western Part of Nigeria. Pakistan Gahlot, M., Bhatt, P., Joshi, J., Fellow, S. R., &
Journal of Nutrition, 6(6), 665-668. Pantnagar, T. (2018). Study on Yield of Plant
Adejuwon, A. A., & Olufunmilayo, O. A. (2009). Extracts Using Different Solvents and Methods. In
Further evaluation of antihyperglycaemic activity Bull. Env. Pharmacol. Life Sci.
of Hunteria umbellata (K Schum) Hallier f. seed Ganong, W. F. (2003). Circulating Body Fluids: Review

------------------------------------------------------------------------------------------------------------------------------------------------------
Anani, Shalem, Akpan & David-Okwu
Nigerian Agricultural Journal Vol. 55, No. 1 | pg. 101
 of Medical Physiology, 21st Edn., Typo Press, and Bioresources, 15(4), 1-9.
Lebanon. Murray, D., Arbuzova, A., Hangyamihalyne, G.,
Hadi, M. Y., Mohammed, G. J., and Hameed, I. H. Gambhir, A., Ben-Tal, N., Honig, B., &
(2016). Analysis of bioactive chemical compounds Mclaughlin, S. (1999). Electrostatic properties of
of Nigella sativa using gas chromatography-mass membranes containing acidic lipids and absorbed
spectrometry. Journal of Pharmacognosy and basic peptides: Theory and Experiment.
Phytotherapy, 8(2), 8-24. Bio/Physical Journal. 17:3176-3188.
Hanna, M., Jaqua, E., Nguyen, V., & Clay, J. B. (2022). Nathar, V., Wagay, N., Varsha, N., Nathar, W., & Wagay,
Vitamins: Functions and Uses in Medicine. N. (2018). Investigations on important secondary
Permanente Journal, 26(2), 89-97. metabolites from aerial parts of Artemisia
Harborne, J. B. (1973). Phytochemical Methods-A absinthium L. using GC-MS.
Guide to Modern Techniques of Plant Analysis, Obadoni, B. O., & Ochuko, P. O. (2002). Phytochemical
Chapman and Hall, London, 49-188 studies and comparative efficacy of the crude
Herrera, E., & Barbas, C. (2001). Vitamin E: Action, extracts of some haemostatic plants in Edo and
metabolism, and perspectives. Journal of Delta States of Nigeria. Global Journal of pure and
Physiology and Biochemistry, 57, 43–56. applied sciences, 8(2), 203-208.
Kennedy, D. O. (2016). B Vitamins and the brain: Okwu, D. E. (2005). Phytochemicals, vitamin and
mechanisms, dose, and efficacy--a review. mineral contents of two Nigerian medicinal plants.
Nutrients, 8(2), 68. doi: 10.3390/nu8020068. International Journal of Molecular Medicine and
Kim, Y. H., Park, S. K., Hur, J. Y., & Kim, Y. C. (2017). Advance Sciences. 1(4):375-381
Purification and characterisation of a major Oshiobugie, M. J., Anthony, D. O., Oladipupo, A. A.,
extracellular chitinase from a biocontrol bacterium, Omosalewa, B. A., Michael, A. O., Olabisi, B. N., &
Paenibacillus elgii HOA73. Plant Pathology Aiyegbeni, M. A. (2019). Phytochemical Analysis
Journal, 33, 318. and Antiplasmodial (curative) Activities of
Lawag, I. L., Nolden, E. S., Schaper, A. A. M., Lim, L. Methanolic Leaf Extract of Morinda lucida (Ewe
Y., & Locher, C. (2023). A Modified Folin- Oruwo) in Male Swiss Mice Infected with
Ciocalteu Assay for the Determination of Total Plasmodium berghei NK65. International Journal
Phenolics Content in Honey. Applied Sciences, of TROPICAL DISEASE & Health, 37(1), 1–13.
1 3 ( 4 ) , 2 1 3 5 . [Link]
[Link] Oshodi A.J.L., Ogungenle H.N., Oladimeji N.O.,
Liu, Q., Meng, X., & Li, Y. (2018). Natural products for Chemical composition, nutritionally valuable
the prevention and management of Helicobacter minerals and functional properties of benniseed
pylori infection. Comprehensive Reviews in Food (Sesanum rediatum), pearl millet (Perntiisetim
Science and Food Safety, 17(4), 937-952. typhoides) and quinoa (Chenopodium quinoa)
Lourenço, S. C., Moldão-Martins, M., & Alves, V. D. flours. International Journal of Food Science &
(2019). Antioxidants of Natural Plant Origins: Nutrition., 1999, 50, 325–331.
From Sources to Food Industry Applications. Oyeleke, G. O., Oyetade, O. A., Afolabi, F., & Adegoke,
Molecules, 24(22), 4132. B. M. (2013). Nutrients, Antinutrients and
Madhu, M., Sailaja, V., Satyadev, T. N. V. S. S., & Physicochemical Compositions of Blighia Sapida
Satyanarayana, M. V. (2016). Quantitative Pulp and Pulp Oil (Ackee Apple). IOSR Journal of
phytochemical analysis of selected medicinal plant Applied Chemistry (IOSR-JAC), 4(1), 05-08.
species by using various organic solvents. Journal Oyeleke, G. O., Salam, M. A., & Adetoro, R. O. (2012).
of Pharmacognosy and Phytochemistry, 5(2), 25- Some Aspects of Nutrient of Seed, Pulp, and Oil of
29. Baobab (Adansonia digitata). Journal of
Malek, S. N. A., Shin, S. K., Wahab, N. A., & Yaacob, H. Environmental Science, Toxicology and Food
(2009). Cytotoxic components of Pereskia bleo Technology, 1, 32-35.
(Kunth) DC. (Cactaceae) leaves. Molecules, 14, Oyeleke, G. O., Sulaimon, W. K., & Adebayo, G. O.
1713. (2011). Effect of Dehulling on the Nutritive and
Mboso, O. E., Eyong, E. U., Odey, M. O., & Osakwe, E. Antinutritive Components of Calabash.
(2013). Comparative Phytochemical Screening of International Journal of Chem., 21, 3.
Ereromastax speciosa and Ereromastax Ozioma, E.O.J., & Chinwe, O.A.N (2019). Herbal
polysperma. Journal of National Production and medicines in African traditional medicine. Herb.
Plant Resources, 3(2), 37–41. Med. 10:191-214.
Mintah, S.O., Asafo-Agyei, T., Ocloo, A., Boakye, Y.D., Pereira, D. M., Patrícia, V. P., Pereira, J. A., and
& Agyare, C. (2019). Medicinal plants for the Andrade, P. B. (2009). Phenolics: From Chemistry
treatment of prevalent diseases. Pharmacognosy & to Biology. Molecules, 14, 2202-2211.
medicinal plants. Intechopen 82049:9-18 Pomeranz, Y.M and Cliffton, E. (2015). Food Analysis:
Morakinyo, A. E., Oladeji, O. A., Fajobi, A. O., Theory and Practice. 3rd edition. An Aspen
Adelowo, J. M., Babarinde, S. O., & Adepoju, A. E. publication.
(2020). Phytochemical analysis and Ren, J., Wang, J., Karthikeyan, S., Liu, H., & Cai, J.
antihyperlipidemic potential of ethanolic extract of (2019). Natural anti-phytopathogenic fungi
Hunteria umbellata seeds. Journal of Pharmacy compound phenol, 2, 4-bis (1, 1-dimethylethyl)

------------------------------------------------------------------------------------------------------------------------------------------------------
Anani, Shalem, Akpan & David-Okwu
Nigerian Agricultural Journal Vol. 55, No. 1 | pg. 102
 from Pseudomonas fluorescens TL-1. International Udinyinwe, C.O and Aghedo, E.S. (2022). Proximate
Journal of Endocrinology, 34: pp. 123-125 Composition, Phytochemical and Antimicrobial
Riaz Hossain, M., Sifat Foysal, M., Ibban, S., & Hazrat Activity of Aqueous and Ethanolic Extracts of
Ali, M. (2023). Phytochemical Screening and Hunteria umbellata on some Clinical Isolates.
Evaluation of In Vitro Thrombolytic, Anthelmintic Journal of Applied Science and Environmental
and Vivo Analgesic Activities of Ethanolic Extract Management, Vol. 26 (1) 31–36.
of Leea AequataL. IOSR Journal Of Pharmacy And V i d h u , V. V. , & E v a n s , D . A . ( 2 0 1 5 ) .
Biological Sciences (IOSR-JPBS) e-ISSN, 18, Ethnoentomological values of Oecophylla
59–66. [Link] smaragdina (Fabricius). Current Science, 109(3),
Salisu, T. F., Fowora, M. A., Yahaya, T. O., Aina, S. A., 572–579.
Thomas, B. T., Ademola, L. A., & Jimoh, P. O. Villamor, E., & Fawzi, W. W. (2005). Effects of vitamin
(2024). Phytochemical screening, gas A supplementation on immune responses and
chromatograph/mass spectrometer (GCMS) correlation with clinical outcomes. Clinical
analysis and molecular toxicological potential of Microbiology Reviews, 18, 446-464.
Hunteria umbellata aqueous fruit extract against Welz, A. N., Emberger-Klein, A., and Menrad, K.
Staphylococcus aureus in accessory gene (2018). Why people use herbal medicine: Insights
r e g u l a t o r s ( A G R s ) . F u t u re J o u r n a l o f from a focus-group study in Germany. BMC
Pharmaceutical Sciences 2024 10:1, 10(1), 1–13. Complementary and Alternative Medicine, 18, 92.
[Link] Zahid, M., Arif, M., Rahman, M.A., Singh, K., &
Sirimanne, S. R., Patterson, D. G., Ma, L., & Justice, J. Mujahid, M. (2017). Solvent extraction and gas
B. (1998). Application of cloud-point extraction- chromatography-mass spectrometry analysis of
reversed-phase high-performance liquid text (Annona squamosal) L. seeds for
chromatography: A preliminary study of the determination of bioactive, fatty acid/fatty oil
extraction and quantification of vitamins A and E in composition, and antioxidant activity. Journal of
human serum and whole blood. Journal of Dietary Supplements, 5:613-623.
Chromatography B: Biomedical Sciences and Zhou, B. L., Chen, Z. X., Du, L., Xie, Y. H., Zhang, Q., &
Applications, pp. 716, 129–137. Ye, X. L. (2011). Allelopathy of root exudates from
Sofowora, A. (1993). Medicinal Plants and Traditional different resistant eggplants to Verticillium dahliae
Medicine in Africa (2nd ed.). Ibadan: Spectrum and the identification of allelochemicals. African
Books Ltd. Journal Biotechnol, pp. 10, 8284.
Sofowora, A., Ogunbodede, E., & Onayade, A. (2013). Zhou, D. D., Luo, M., Shang, A., Mao, Q. Q., Li, B.Y.,
The role and place of medicinal plants in the Gan, R.Y., & Li, H. B. (2021). Antioxidant Food
strategies for disease prevention. African Journal of Components for the Prevention and Treatment of
Traditional, Complementary and Alternative Cardiovascular Diseases: Effects, Mechanisms,
Medicines, 10(5), 210-229. and Clinical Studies. Oxidative Medicine and
Trease GE, Evans WC (1989). Pharmacognosy 2nd Cellular Longevity, 2021, Article ID 6627355, 17
Edn. Braille Tiridel and Macmillan Publishers. pages.
Table 1: Proximate composition of ethanol extract of H. umbellata seed
Proximate composition Values (%)
Moisture content (dry wt.) 16.20±0.16
Crude protein 13.92±0.01
Lipid 2.59±0.02
Ash 2.53±0.06
Available carbohydrate 56.73±0.24
Fibre 8.03±0.41
Calorific Value (KJ/100g) 1277.43±3.18
Values are Mean ± SD of triplicate determination: wt=weight
Table 2: Qualitative phytochemical composition of ethanol extract of H. umbellata seeds
Phytochemical composition Result
Alkaloids +
Flavonoids ++
Glycosides -
Reducing sugar +
Saponins ++
Steroids -
Tannins +
Terpenoids -
Phytates +
Oxalates +
Cyanogenic glycosides (mg/L) +
(+): present, (++): strong presence
------------------------------------------------------------------------------------------------------------------------------------------------------
Anani, Shalem, Akpan & David-Okwu
Nigerian Agricultural Journal Vol. 55, No. 1 | pg. 103
Table 3: Quantitative phytochemicals composition of ethanol extract of H. umbellata seeds
Phytochemicals Values (mg/100g)
Alkaloids 1.80±0.28
Flavonoids 383.50±0.24
Reducing sugar 26.16±0.52
Saponins 150.27±0.11
Tannins 14.02±0.01
Phytates 0.26±0.05
Oxalates 10.42±0.17
Cyanogenic glycosides (mg/L) 6.21±0.04
Each value is expressed as mean ± standard deviation of triplicate determinations.
Table 4: Chemical composition of ethanol seed extract of Hunteria umbellata, identified using Gas
Chromatography-Mass Spectrometry (GC-MS)
Pk Retention Area Library/ID Molecular Quali
no time % Formula ty
1 3.665 0.31 Hexamethyl-Cyclotrisiloxane C6H18O3Si3 72
2 8.285 0.51 2-methyl-Benzaldehyde C8H8O 96
3 9.835 0.74 3,5-dimethyl-Benzaldehyde C9H10O 95
4 9.665 1.27 3,5-dimethyl-Benzaldehyde C9H10O 94
5 11.44 0.60 4-Butyl-5-(3-methylbutyl)-6-(1-methyl phenyl)-2H- C9H13NO2 25
pyran-2-one
6 11.975 37.72 Phenol, 2,4-bis(1,1-dimethylethyl) C14H22O 97
7 12.511 10.75 4-Amino-7-diethylamino-chromen-2-one C13H16N2O2 64
8 13.018 0.98 Z-11-Pentadecenal C15H28O 93
9 13.159 0.44 Oleic Acid C18H34O2 90
10 14.06 6.13 cis-Vaccenic acid C18H34O2 94
11 14.201 1.36 18-Nonadecenoic acid C19H36O2 84
12 14.398 2.65 Z-(13,14-Epoxy) tetradec-11-en-1-ol acetate C16H28O3 91
13 14.623 2.09 Cyclopentadecanone, 2-hydroxy- C15H28O2 92
14 14.877 1.41 Butyl 9-octadecenoate or 9-18:1 C22H42O2 80
15 15.159 9.47 trans-13-Octadecenoic acid C18H34O2 91
16 15.553 2.94 -Octadecene, (E)- C18H36 83
17 16.088 3.17 1-(ethenyloxy)-Octadecane, C20H40O 83
18 16.37 1.41 1-Nonadecene C19H38 89
19 18.088 8.71 9-Octadecenamide, (Z)- C18H35NO 64

Table 5: The fat-soluble vitamin content of Hunteria umbellata seeds

Vitamins Values
Vitamin A (iU /100g) 21.95±0.01
Beta Carotene (mg/100g) 0.41±0.00
Vitamin D (IU/100g) 8.30±0.14
Vitamin E (mg/100g) 1.25±0.07
Vitamin K (IU/100g) 135.00±0.14
Values presented are mean ± Standard deviation triplicate determination.

Table 6: Vitamin-B complex content of ethanol extract of Hunteria umbellata seeds

Peak No. Peak ID Ret Time Height Area Conc (mg/100g)
2 Niacin 0.473 320.906 436.28 0.0481
4 Thiamine 0.59 299.143 404 0.0446
6 Folic acid 1.123 1165 3059.9 0.3375
7 Biotin 1.515 7552.552 79686.2 8.789
10 Pyridoxine 2.857 620.085 1835.821 0.2025
13 Riboflavin 3.973 16790.8 802520.8 88.5138
15 Cobalamin 6.032 647.92 5219.2 0.5757

------------------------------------------------------------------------------------------------------------------------------------------------------
Anani, Shalem, Akpan & David-Okwu
Nigerian Agricultural Journal Vol. 55, No. 1 | pg. 104
Table 7: Selected Minerals composition of ethanol extract of Hunteria umbellata seeds
Minerals Values (mg/L)
Calcium (Ca) 7.74±0.08
Phosphorous (P) 3.31±0.01
Iron (Fe) 3.51±0.02
Magnesium (Mg) 4.22±0.21
Manganese (Mn) 5.47±0.44
Sodium (Na) 80.59±0.66
Potassium (K) 9.37±0.22
Zinc (Zn) 7.08±0.11
Values presented are mean ± Standard deviation of triplicate determinations.

------------------------------------------------------------------------------------------------------------------------------------------------------
Anani, Shalem, Akpan & David-Okwu
Nigerian Agricultural Journal Vol. 55, No. 1 | pg. 105