Volume 8, Issue 9, September – 2023 International Journal of Innovative Science and Research Technology

ISSN No:-2456-2165

Phytochemical Characterization of Dichloromethane

Stem Bark Extract of Piliostigma thonningii (Schum.)
Farida Moraa Okioga, Msc. Biotechnology, Mathew P. Ngugi
https://orcid.org/0000-0002-6205-1394 Ph.D. (Biochemistry and Molecular Biology)
Department of Biochemistry, https://orcid.org/0000-0002-0226-9092
Microbiology and Biotechnology Kenyatta University. Department of Biochemistry, Microbiology and
Biotechnology Kenyatta University

Huxley Mae Makonde

https://orcid.org/ 0000-0002-0163-9303
Department of Pure and Applied Sciences,
Technical University of Mombasa

Abstract:- Piliostigma thonningii (Schum.) commonly widespread significance (Goni et al., 2021). Herbal medicine
referred to as monkey bread belongs to Fabaceae family offers an alternative avenue for healthcare as the global
and is used in the tropics for medicinal and nutritional burden of disease continues to grow. Search of new
uses. The plant is utilized for management of human therapeutics from plant source is an active area of research as
ailments among them rheumatism, malaria, fever, snake new treatment intervention is required to grapple with
bites and dysentery. It is widely acknowledged that plant existing and emerging threats to human health (Reza et al.,
pharmacological activities are driven by their secondary 2021). Past evidences have indicated that plant species have
metabolites. Identification of the metabolites has led to numerous pharmacological properties and a large number of
drug discovery and leads for designing of novel them are already being applied for treatment of various
compounds to enhance human health. Though P. human ailments including cancer, diabetes, pain, arthritis,
thonningii is endowed with numerous bioactivities, there malaria (Martínez et al., 2020; Reza et al., 2021).
is minimal literature detailing the phytochemicals
responsible for its therapeutic efficacies. For that reason, The enormous health benefits associated with herbal
the present embarked on quantitative phytochemical produced is accredited to the phytochemicals they contain
characterization of DCM stem bark extract of P. (Reza et al., 2021). The phytochemicals have a wide array of
thonningii. The analysis was carried out using gas applications including nutraceuticals, drug manufacturing
chromatography-mass spectrometry. The results showed industries, food supplements, medicine and chemical leads
that the extract contained various classes of for synthetic drug design (Pollio et al., 2016; Ibrahim et al.,
phytocompounds including flavonoids, phenolics, 2023). The main phytochemicals present in plants include
phytosterols and fatty acids. The most abundant alkaloids, flavonoids, phenolic acids, terpenoid, sterols and
compounds found in the extract were oleic acid and triterpenes (Reza et al., 2021). Alkaloids are known have
squalene whereas valencene and α-cedrene were the least analgesic and antimicrobial properties (Saranraj and
concentrated. The compound identified in the extract Sivasakthi, 2014), glycosides can moderate blood pressure
have been reported to possess numerous pharmacological (Han et al., 2007), phenolic compounds are known to possess
efficacies such as analgesic, anti-inflammatory, anticancer various biological activities including apoptosis, anti-aging,
and antidiabetic activities. Thus, the promising anti-inflammation, anti-oxidant, anti-atherosclerosis,
therapeutic potential of P. thonningii is attributed to these inhibition of angiogenesis, (Shrestha et al., 2015) and others,
phytocompounds. The study recommends further saponins have anti-inflammatory and antibacterial potency
research to isolate and purify the activity ingredient from (Chang et al., 2023), terpenoids exhibits antimicrobial and
P. thonningii as well as determine their mechanism of antimalaria activities (Mahizan et al., 2019).
action.
Though there is enormous literature regrading
Keywords:- P. thonningii, Phytochemicals, gas phytochemical composition of medicinal plants, little is
chromatography-mass spectrometry. known regarding the phytochemistry of Piliostigma
thonningii. Consequently, this study aimed to investigate the
I. INTRODUCTION chemical constituents of dichloroethane (DCM) stem bark
extract of P. thonningii as a potential source of lead
Botanicals with therapeutic properties have been compounds for developing safe, efficacious, and affordable
utilized in health care since the birth of human civilization therapeutics. Piliostigma thonningii (Fig 1), an evergreen
(Moni et al., 2021). Globally, numerous efforts have been shrub, belongs to Fabaceae family, sub family
enlisted to investigate the effectiveness of plant-based Caesalpinioideae. It natively grows in the tropics of Africa.
medicines with some leading to discovery of drugs of Its common names include monkey bread, camel's foot, wild

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Volume 8, Issue 9, September – 2023 International Journal of Innovative Science and Research Technology
ISSN No:-2456-2165
bauhinia, (English) and mchikichi (Swahili) (Orwa et al., Department of Plant Sciences, Kenyatta University (Voucher
2009). The plant bears hard and hairy fruits. Its fruit pulp is Specimen Number GM001/2017). The samples were cleaned
rich in organic acids, proteins, fibre, vitamins, calcium with tap water, rinsed with distilled water, then chopped into
oxalates, starch and essential oils (Orwa et al., 2009). Its small pieces. After this, the pieces were shade dried on a
traditionally used to treat and manage ulcers, pain, wounds, laboratory bench. After completely drying, they were milled
arthritis, fever, malaria, sore throat, diarrhea, gingivitis, into powder with an electric mill. The resulting stem bark
respiratory conditions (Ighodaro and Omole, 2012; Afolayan powder was packaged into labeled khaki sack and stored at
et al., 2018). Its crude extracts have been demonstrated to 25 ± 2 oC.
exert antibacterial (Akinpelu and Obuotor, 2000), anti-
lipidemic (Ighodaro and Omole 2012), anti-inflammatory B. Preparation of Plant Extracts
(Ibewuike et al., 1997) and anti-helminthic (Asuzu and Onu, The powdered plant material (200 g) was soaked in
1994) activities. 750 ml of analytical-grade dichloromethane (DCM) in a
conical flask and covered with aluminum foil. The mixture
was left to stand for 48 hours during which it was shaken
periodically and stirred with a glass rod. Thereafter, the
extract was filtered through Whatman filter papers (No.1)
and concentrated in vacuo using a rotary evaporator at 28 0C.
The resulting crude extract was transferred into clean, dry,
pre-weighed glass bottles and completely air-dried at room
temperature.

C. Phytochemical analysis of the DCM stem bark extract of

P. thonningii
The bioactive constituents of DCM stem bark extract of
P. thonningii were characterized using Gas Chromatography
- Mass Spectrometry (GC-MS) following published protocols
(Hossain et al., 2020; Ibrahim et al., 2021).

III. RESULTS AND DISCUSSION

 Phytochemistry of P. thonningii
Fig 1 Piliostigma thoningii (Schumach) The results revealed various phytocompounds of
various classes including fatty acids, triterpenoids,
II. MATERIALS AND METHODS phytosterols, phenols, whereby oleic acid (218.60±15.6 ng/g)
and 1-nonadecene (33.34±2.38 ng/g), and squalene
A. Collection of Plant Material (31.45±2.25 ng/g) were the most abundant and carvacrol
Stem barks of P. thonningii were collected from Mbeere (1.37±0.10 ng/g), valencene (0.72±0.05 ng/g), and α-cedrene
North sub-County, Embu County in Kenya. The plant (0.69±0.05 ng/g) were the least abundant (Table 1).
materials were authenticated by a taxonomist at the

Table 1: Phytochemical composition of the DCM stem bark extract of P. thonningii

Compound name Molecular formula Chemical class Concentration (ng/g)

Oleic Acid C18H34O2 Fatty acid 218.60±15.6
Squalene C30H50 Triterpene 31.45±2.25
Alpha Amyrin C30H50O Triterpenoid 28.88±2.06
Decosanoic acid C22H44O2 Fatty acid 23.61±1.69
n-Hexadecanoic acid C16H32O2 Fatty acid 21.88±1.56
Gamma Tocopherol C28H48O2 Phenolic 9.52±0.68
Gamma Sitosterol C29H50O Phytosterol 5.85±0.42
Campesterol C28H48O Phytosterol 5.50±0.39
Palmitoleic acid C16H30O2 Fatty acid 4.28±0.306
4-(2,4,4-trimethylpentan-2-yl) phenol C14H22O Phenolic 2.38±0.17
Pentadecanoic acid C15H30O2 Fatty acid 2.27±0.162
Stigmasterol C29H48O Phytosterol 1.49±0.12
Carvacrol C10H14O Phenolic 1.37±0.10
Valencene C15H24 Sesquiterpene 0.72±0.05
α-cedrene C15H24 Sesquiterpene 0.69±0.05

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Volume 8, Issue 9, September – 2023 International Journal of Innovative Science and Research Technology
ISSN No:-2456-2165
Other studies have reported similar findings on the IV. CONCLUSION
classes of secondary metabolites present in P. thonningii.
Moriasi et al. (2020) reported that P. thonningii methanolic The findings of the present study demonstrated that P.
and aqueous stem bark extracts contains glycosides, phenols, thonningii DCM extract contains constituents with promising
steroids, saponins, and flavonoids. Further, flavonoids, therapeutic potential. Multiple previous studies confirmed the
saponins and terpenes were confirmed resent in ethanol therapeutic activities of the identified compounds. Thus, the
extract of P. thonningii extract (Egharevba and Kunle, 2010). therapeutic potential associated with P. thonningii is
Phytochemical analysis of P. thonningii roots showed it mediated by its phytochemical constituents. Based on the
contained alkaloids, flavonoids, terpenoids, tannins, phenols, study findings P. thonningii may provide dependable
saponins and steroids (Alagbe, 2019). reservoir for therapeutic agents.

Bioactivities of some of the compounds identified in P. DECLARATIONS

thonningii have been reported in literature. Fatty acids such
as oleic and palmitoleic acid has been reported to possess  Author Contribution Statement
antioxidant, anti-inflammatory and anti-Alzheimer activities All the authors conceived the research idea. Farida
(Mazouz et al., 2020; Agidew et al., 2021; Santa-María et al., Moraa Okioga performed the experiments, analysed the data,
2023). Squalene, a triterpene, has been show to exhibit and drafted the manuscript under the supervision of Dr
cardioprotective, anticancerous and antioxidant properties Mathew Piero Ngugi and Dr Huxley Mae Makonde. All the
(Kumar et al., 2023). Triterpenoid alpha amyrin, has been authors reviewed and approved the final version of the
indicated to exert anti-microbial, antitumor, anti- manuscript for submission.
inflammatory, anxiolytic, and hepatoprotective potencies
(Viet et al., 2021).  Funding Statement
This research did not receive funding
Gamma Tocopherol, a phenolic compound, has been
reported to possess antioxidant, anti-inflammatory,  Competing Interest Statement
antidiabetic, anticancer, anti-nociceptive activities (Ebrahimi The authors declare no conflict of interest.
et al., 2021; Zakaria et al., 2021). Another phenolic
compound, carvacrol has been revealed to possess a wide ACKNOWLEDGMENT
array of bioactivities among them; anti-inflammatory,
immunomodulation, antioxidant, antifungal and antibacterial The authors appreciate the Department of Biochemistry,
efficacies (García-Risco et al., 2011). Microbiology and Biotechnology, Kenyatta University for
providing the infrastructure to carry out the study.
Valencene and α-cedrene, sesquiterpenes, have been
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