Vol 3, Issue 2, 2024

ISSN: 2790-9522 (Print) | 2790-9530 (Online)

Website: https://journals.jozacpublishers.com/ajbcps/index

Biological and chemical characterization of Acalypha wilkesiana leaf

extracts

Olajide Ebenezer Falayi 1, Olakunle Ayodeji Fatokun 1, Adewale Elijah Fadeyi 1

1Sheda Science and Technology Complex, Sheda, Abuja, Nigeria, wale.fade@gmail.com

*Correspondence: wale.fade@gmail.com
Received: June 21, 2024 | Accepted: July 29, 2024 | Published: August 16, 2024

Abstract
According to African folklore, Acalypha wikesiana is known to possess medicinal properties. A study
was conducted to evaluate the presence of chemical compounds and biological properties in the non-
polar and crude extracts of Acalypha wikesiana. To obtain the crude extract, the air-dried pulverized
leaf of the plant was extracted with 95% ethanol and hexane. Some microbial strains associated with
dermophytic diseases, including Staphylococcus aureus, Escherichia coli, Aspergillus niger,
Epidermatophyton Sp, Trichophyton rubrum and Candida tropicalis were inhibited by the crude
ethanol extract. No activities were observed against Methylene-resistant Staphylococcus aureus,
Streptococcus pyogenes, Microsporum gypseum, Microsporum canis, Aspergillus fumigates, and
Candida albicans. In the DPPH free radical scavenging assay, the crude ethanol extract exhibited
acceptable robust antioxidant activity between 0.1 and 0.9 mg/mL concentration. In the following GC-
MS analysis, yellow oil was obtained from the crude extract by column chromatography, which
revealed the presence of some compounds, mostly terpenoids and fatty acid esters. Medicinal
properties of Acalypha wikesiana leaf extracts may be attributed to these compounds.

Keywords: Acalypha wikesiana, Phytochemical, Antioxidant, DPPH, GC-MS analysis

1. Introduction
The rising demand for medicinal plants has increased in recent years, driven by the hope of
discovering new, affordable pharmaceuticals (Jamshidi-Kia et al., 2017; Zahra et al., 2020). This
renewed interest in herbal medicine has led to a focus on the study of botanical plants, which contain
a wide range of chemicals with potential uses for human health (Ganjhu et al., 2015).

2. Literature review
Plants are rich in secondary phytochemicals, including minerals, alkaloids, flavonoids, glycosides,
phenols, saponins, and terpenoids (Shakeri et al., 2012; Uttu et al., 2015). Phytotherapy plays a crucial
role in modern medicine, recognized for its potential health benefits and integral to various
applications, including medical, nutraceutical, pharmaceutical, and cosmetic uses (Swamy et al.,
2019). A variety of phytopharmaceutical properties, including anti-inflammatory, anti-thrombogenic,

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diabetic, anti-cancer, and neuroprotective properties, have been shown to work in vitro and in
animals (Ezenyi et al., 2016; Gullon et al., 2017). Some of these medications are derived from plants,
including aspirin, which comes from the bark of willow trees (Salix alba), and quinine, which is
derived from Cinchona. The use of herbal remedies can not only cure disease but also help with
wellness promotion, upkeep, and prevention (Murage et al., 2021; Lin et al., 2021). Many plants native
to Nigeria have been studied for their potential therapeutic uses. Acalypha wilkesiana, commonly
referred to as "Irish petticoat," is a visually striking plant of the Euphorbiaceae family, prized for its
colorful leaves and widely cultivated in tropical and subtropical areas for its decorative appeal and
belongs to the Euphorbiaceae family. It is widely planted in tropical and subtropical locations due to
its high decorative value because of its strikingly coloured leaf (Kingsley & Marshal, 2014). Acalypha
wilkesiana has been used for millennia to treat diabetes, cancer, hypertension, epilepsy, infertility, and
arthritis (Mustafa, 2014) The leaves of A. wilkesiana are used to cure a variety of skin conditions in
Northeastern Africa and the Southwest of Nigeria (Adesina et al., 2000; Madziga et al., 2010; Sharma
et al., 2014). Acalypha wilkesiana leaf has been shown to reduce the prevalence of pathogens such as
Staphylococcus aureus, Staphylococcus pyogenes, Klebsiella aerogenes, and Escherichia coli (Oluduro et al.,
2011; Olude et al., 2022). The goal of this study was to use GC-MS to characterise the phytochemical,
antioxidant and antibacterial characteristics of Acalypha wikisiana leaf extracted in hexane and ethanol.

Figure 1 : Plate 1 - Calotropis procera

3. Research methodology
3.1. Collection of Acalypha wikesiana
The fresh leaf of Acalypha wikesiana was collected from Sheda, Kwali Local Government of Abuja, and
authenticated at the National Institute for Pharmaceutical Research and Development NIPRID,
Abuja, with voucher number (NIPRD/H/7343). Hexane ethanol and methanol are the main solvents
used. All the reagents and solvents used in this investigation were all of standard grade. Assays
involving DPPH (2, 2-diphenyl-1-picrylhydrazyl) was used to measure the antioxidant activity of the
extract. MHA (Mueller Hinton agar) and SDA (Sabourad dextrose agar) were used for the assay. An
antimicrobial test was performed on organisms obtained from the Department of Medical
Microbiology at Ahmadu Bello University Teaching Hospital in Zaria, Kaduna state, Nigeria.
Staphylococcus aureus, E. coli, Methicillin-resistant Staphylococcus aureus, Streptococcus pyogenes, Candida
albicans, Candida tropicalis, Aspergillus nigre, Epidermatophyton sp., Microsporum gypseum, Microsporum
canis, and Trichophyton rubrum Ciprofloxacin, Fluconazole, and Fulcin were employed as the standard
antibiotic agents. Hammer mill (TRP80 Rotary vacuum evaporator (BUCHI), UV spectrophotometer

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(CECIL), and gas column and mass spectrometry GC-MS (Thermo-Scientific Trace GC ULTRA
system) were included in this study.

3.2. Extraction of plant material

Air-dried crispy leaf samples of Acalypha wikesiana (100g) were ground to powder using a hammer
mill, TRP80, and kept before use. The powdered leaf material was separately extracted using the
maceration method for 48 hours with 2.5 liters of hexane and 2.5 liters of 95% ethanol respectively
and later filtered. The resulting extracts were concentrated using a rotary evaporator at 40°C,
followed by air drying to yield the resultant extracts of hexane and ethanol.

3.3. Phytochemical screening

Ethanol crude leaf extract was subjected to phytochemical screening to identify major constituents
using standard methods (Trease & Evans, 2002; Singh et al., 2009).

3.4. DPPH radical scavenging assay

Using a UV-visible spectrophotometer at 517 nm, the ethanol extract's ability to neutralize free
radicals against DPPH (Sigma-Aldrich) was evaluated. A slightly modified method from the one that
was first outlined by (Fadeyi et al., 2022) was employed. Using the following formula, the radical
scavenging activity (RSA) was determined as the percentage inhibition of DPPH discoloration:

Ab is the absorption of the blank sample (without the extract) and A a is the absorption of the extract.

3.5. Antimicrobial screening

Acalypha wilkesiana crude ethanol extract was utilized to screen human pathogenic microorganisms.
Antimicrobial activity was evaluated using the agar well disc diffusion method (Deeni & Sadiq, 2002).
Crude extract (0.1 g) was dissolved in 10 ml DMSO to yield a 10 mg/ml solution. The bacterial
inoculation medium, known as Mueller-Hinton Agar (MHA), and the fungal inoculation medium,
Sabouraud Dextrose Agar (SDA) plates were inoculated with 0.1 ml inoculum, then 0.1 ml crude
extract was added to wells. using a sterile 6 mm corn borer. Following a 24-hour incubation period
at a temperature of 37°C, the plates were subjected to examination in order to identify zones of
inhibition. The determination of the lowest inhibitory concentration of the extract was conducted
using the broth dilution method. A volume of 10 mL of Mueller Hinton broth was dispensed into
individual test tubes. The broths were sterilized at 121°C for 15 minutes and then cooled. The
concentration was determined using a McFarland 0.5 turbidity standard. The test microorganism was
inoculated and incubated at a temperature of 37°C for 6 hours following the production of normal
saline. Subsequently, 10 mL of the saline solution was transferred into a sterile test tube. The test
microbe was diluted in normal saline until it reached the same level of turbidity as Mc-scale Farland's
standard, as determined through visual comparison (Ali et al., 2017).

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3.6. GC-MS analysis of extractives

Hexane oil extracts from fresh leaves were analyzed by GC-MS (7820) and identified by comparing
mass spectral data to the NIST 14 Library and literature (Derwich et al., 2010; Palá-Paúl et al., 2012).

4. Results
Table 1: Phytochemical analysis of Acalypha wilkesiana leaf extracts

hyto-constituents Presence/absence

Phenolics +

Flavonoids +

Tannins +

Saponnins +

Glycosides +

Terpenoids +

Steroids +

Alkaloids +

Carbohydrates +

Resin +

Table 2: Antioxidant activity of Acalypha wilkesiana ethanol leave extract

Conc.(mg/mL) % Inhibition A.W % Inhibition (Vit C)

0.1 73.72 60.38

0.3 62.18 72.50

0.5 44.00 77.69

0.7 32.97 77.88

0.9 16.84 81.54

Figure 2: Antioxidant activity of A. wilkesiana

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Table 3: Zone of inhibition of the extract on microbes

Test A.w CIP FLU FLC MIC MBC/MFC

organisms (mm) (mm) (mm) (mm) (mg/mL) (mg/mL)

S. aureus 24.0 0.00 0.00 0.00 2.50 5.00

E. Coli 21.0 38.0 0.00 0.00 2.50 10.00

0 0

A. niger 21.0 0.00 0.00 29.00 2.50 10.00

E. sp 23.0 0.00 0.00 28.00 2.50 10.00

T. rubrum 25.0 0.00 0.00 27.00 2.50 5.00

C. tropicalis 27.0 0.00 32.00 0.00 1.25 5.00

M. R. St. 0.00 32.0 0.00 0.00 0.00 0.00

aureus 0

S. pyogenes 0.00 30 0.00 0.00 0.00 0.00

.00

M.gypseum 0.00 32.0 0.00 0.00 0.00 0.00

M.canis 0.00 32.0 0.00 0.00 0.00 0.00

A.fumigatus 0.00 0.00 0.00 0.00 0.00 0.00

C.albicans 0.00 0.00 34.00 0.00 0.00 0.00

Key: CIP: Ciprofloxacin, FLU = Fluconazol, FLC = Fulcin

Figure 3: GCMS spectral of refined oil

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Table 4: GC-MS characterization of oil extract of A. wilkesiana leaf

RT % Compounds

Area

3.468 0.191 Cyclopentanol, 3-methyl-

3.670 0.146 Benzene, 1,3-dimethyl-

3.988 0.024 Cyclohexanol

4.346 0.039 Oxirane, 2-methyl-2-pentyl-

4.577 0.043 (R)-(-)-2-Methyl-2,4-pentanediol

4.710 0.060 Isothiazole, 3-methyl-

4.975 0.015 Benzene, 1-ethyl-3-methyl-

5.074 0.008 2-propenamide, 2-methyl-N-[3-[(phenylsulfonyl)amino] propyl]-

5.224 0.009 Chloromethyl 2-chloroundecanoate

5.316 0.009 1-Undecene, 5-methyl-

5.409 0.033 Mesitylene

5.894 0.113 D-Limonene

6.206 0.049 Octadecane, 1-(ethenyloxy)-

6.766 0.029 Undecane

7.014 0.021 9-Borabicyclo [3.3.1] nonane, 9-hydroxy-

7.182 0.022 Benzene, 2-butenyl-

7.615 0.011 Cyclopropane, 1-hexyl-2-propyl-, cis-

7.840 0.049 Butanedioic acid, diethyl ester

8.013 0.078 Dodecane

8.285 0.012 Pentadecanoic acid, 14-bromo-

8.672 0.014 1, E-11, Z-13-Octadecatriene

8.885 0.008 2-Piperidinone, N-[4-bromo-n-butyl]-

9.088 0.017 4-Methyl-dodec-3-en-1-ol

9.203 0.018 Tridecane

9.324 0.013 Cyclohexanol, 5-methyl-2-(1-methylethenyl)-

9.503 0.011 Naphthalene, 1,2,3,4-tetrahydro-1,4-dimethyl-

9.936 0.026 Naphthalene, 1,2,3,4-tetrahydro-1,1,6-trimethyl-

10.318 0.106 Tetradecane

10.699 0.015 2,6,10-Dodecatrien-1-ol, 3,7,11-trimethyl-

10.982 0.052 Dodecane, 2,6,11-trimethyl-

11.317 0.034 trans- beta –Ionone

11.507 0.048 Nonanoic acid, 9-oxo-, ethyl ester

11.756 0.049 Furan, 2,5-dibutyl-

11.860 0.074 2(4H)-Benzofuranone, 5,6,7,7a-tetrahydro-4,4,7a-trimethyl-, (R)-

12.160 0.120 Decanoic acid, ethyl ester

12.304 0.096 5-Eicosene, (E)-

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12.870 0.046 2,3-Dimethyl-8-oxo-non-2-enal

12.986 0.053 2-Cyclohexen-1-one, 4-(3-hydroxy-1-butenyl)-3,5,5-trimethyl-, [R-[R*,R*-(E)]]-

13.107 0.010 Cyclopentane, 1-butyl-2-pentyl-

13.240 0.050 Diethyl azelate

13.373 0.062 Pentadecane, 2,6,10,14-tetramethyl-

13.483 0.023 1-Octadecene

13.604 0.042 1-Hexadecanol, 2-methyl-

14.014 0.095 Tetradecanoic acid

5. Discussions
The ethanol extract of the plant contains notable chemical compounds that are responsible for its
various pharmacological effects (Table 1). Flavonoids, terpenoids, tannins, alkaloids, cardiac
glycosides, and carbohydrates. Terpenoids are the primary constituents of essential oils which are
vital as aromatherapy (Hassanpour et al., 2020; Fongang et al., 2021). Due to their mild astringency,
tannins are important secondary metabolites that have the potential to be antibacterial agents
(Clinton, 2009; Voon et al., 2011; Stephane & Jules, 2020). Flavonoids, which are plant pigments, are
potent antioxidants that help to prevent oxidative diseases like cardiovascular disease and certain
types of cancer (Ogbuehi et al.,2014; Panche et al, 2016). Glycosides offer several medical benefits,
including antioxidant, anti-inflammatory, and anti-diabetic properties, highlighting their potential in
disease prevention and treatment (Tran et al., 2020). A broad class of substances recognized for their
detergent and foam-forming abilities is saponins (Bezerra et al, 2018).
Free radicals, charged molecules produced by biological metabolism, could harm healthy cells
Antioxidants have become linked to good health due to their capacity to mitigate some of their
harmful effects (Kehrer & Klotz, 2015). In the fight against cancer, heart disease, stroke, and other
immune-compromising conditions, eliminating free radicals may be helpful (Lobo et al., 2010). The
linear regression coefficients of extracts and ascorbic acid in the concentration versus percent analysis
were 0.995 and 0.8326, respectively. The value of the IC50 = 0.444. It has strong antioxidant property
that is due to flavonoids and tannins (Clinton,2009), which are crucial in absorbing and neutralizing
free radicals (Table 2). These phytochemicals have also been linked to lower mortality rates for a
variety of different human illnesses (Panche et al., 2016; Hassanpour et al., 2011). The crude ethanol
leaf extracts' bioactivities were examined against many pathogenic bacteria and fungi associated with
skin and gastrointestinal disorders (Table 3). The plant extract exhibited significant antimicrobial
activity against a variety of test organisms, including Staphylococcus aureus, Escherichia coli, Aspergillus
niger, Enterobacter sp., Trichophyton rubrum and Candida tropicalis with minimum inhibitory
concentrations (MICs) ranging from 1.25 to 2.50 mg/mL and minimum bactericidal/fungicidal
concentrations (MBCs/MFCs) of 1.25 to 2.50 mg/mL. S. aureus and E. coli are the most common causes
of disease and death in those with impaired immune systems associated with gastrointestinal
disorders while A. niger, E.sp., T. rubrum, and C. tropicalis has been associated to hospital
environments (Amenu, 2014; Nwachukwu et.al.,2016). The ethanol crude leaf extract activities agree
with earlier works by (Gotep et al., 2010).

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The leaf extract was relatively inactive against the test organisms such as M. R. Staphylococcus, C.
albicans, S. Pyogenes, M.gypseum, M.canis and A.fumigatus. According to the results of the extract's
phytochemical analysis, flavonoids and tannins were found. The observed bioactivity is a result of
the flavonoids and tannins that constitute the plant extract. (Yu et al., 2021 Alkaloids have inspired
antibacterial drugs and served as crucial scaffolds for their development (Cushnie, et al., 2014). The
hexane leaf extract was analyzed by GC-MS, revealing unique pharmacological characteristics due to
the presence of fatty acid esters and derivative. Hence the plant contain known chemicals with
distinct pharmacological potencies (Onocha & Olusanya, 2021). Acalypha wilkesiana's leaf extracts
were found to possess antioxidant and antibacterial properties, showcasing its potential as a natural
antimicrobial agent, and GC-MS analysis revealed its phytochemical composition.

6. Recommendations
Assessing the biological and chemical characteristics of Acalypha wilkesiana leaf extracts have
enhances our understanding of its therapeutic potential, hence reaveling the chemical composition
of the plant and prospective pharmaceutical uses.

7. Conclusion
Among the main constituents of Acalypha wilkesiana oil leaf extracts are long-chain hydrocarbons,
fatty acids, and their derivatives, in addition to being antibacterial and antifungal, these substances
are known to have a variety of medicinal properties. This study supports the plant's historical use as
a remedy for various ailments since it contains a number of phyto-constituents.

8. Acknowledgments
The authors thank SHESTCO for access to the Chemistry Advanced Research Centre laboratory
facilities, which facilitated this study.

ORCID
Olajide Ebenezer Falayi https://orcid.org/0000-0002-3463-3899
Fatokun Ayodeji Olakunle https://orcid.org/0000-0001-6581-9699
Adewale Elijah Fadeyi https://orcid.org/0000-0001-7152-7131

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