Phytochemical Screening and Antibacterial Activity Studies on the Crude Leaf Extract of Solanum sisymbriifolium: Traditional Ethiopian Medicinal Plant

Abstract

Ethiopians have used medicinal plants for centuries. In some cases, it is important and the only treatment source. However, these plant species have not been fully studied. In addition, society is inevitably losing knowledge of traditional medicinal plants as society adopts new lifestyles. Consequently, the objective of this research was to determine the phytochemical components and antibacterial activity of Solanum sisymbriifolium, a traditional medicinal plant used in Ethiopia to treat arthritis. Phytochemical analyses were performed on the leaf extracts to identify the bioactive constituents. The results of this study indicated that the plant contains carbohydrates, phenols, flavonoids, alkaloids, proteins, steroids, saponins, and terpenoids. Tannins and anthraquinones were absent. Alkaloids and terpenoids' presence in the leaves of these plants is a potential bioactive for bacterial inhibitors. At optimal conditions (62°C, 72 hr, and 1 mm particle size), the maximum extraction yield is 38:538:5 ± 1:15%. Crystals obtained from aqueous methanol extracts were subjected to FT-IR, and the compound spectrum showed a characteristic absorption band for the N-H group at 3500 cm-1 and 1700 cm-1 for the C = O group, and the medium intensity at 1236 cm-1 indicates a CO stretching. Sharp absorption at 707 cm-1 is inductive for = C-H bending. According to agar disc diffusion tests, plant extracts of 50 mg/mL produced 14.04 mm growth inhibition zones of Bacillus subtilis. Phytochemical and antibacterial studies of Solanum sisymbriifolium indicated that the plant is a source of highly valued compounds for the preparation of medications.

Full text

Research Article
Phytochemical Screening and Antibacterial Activity Studies on
the Crude Leaf Extract of Solanum sisymbriifolium: Traditional
Ethiopian Medicinal Plant
Gebrihans Haile Gebrewbet
1
and Abadi Gebreyesus Hndeya
2
1
School of Chemical and Bio Engineering, Dire Dawa University Institute of Technology, Dire Dawa 3000, Ethiopia
2
Department of Chemical Engineering, Mekelle Institute of Technology-Mekelle University, Mekelle 7000, Ethiopia
Correspondence should be addressed to Gebrihans Haile Gebrewbet; gebrihanss26@gmail.com
Received 20 March 2023; Revised 28 April 2023; Accepted 10 May 2023; Published 23 May 2023
Academic Editor: Jian Wu
Copyright © 2023 Gebrihans Haile Gebrewbet and Abadi Gebreyesus Hndeya. This is an open access article distributed under the
Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided
the original work is properly cited.
Ethiopians have used medicinal plants for centuries. In some cases, it is important and the only treatment source. However, these
plant species have not been fully studied. In addition, society is inevitably losing knowledge of traditional medicinal plants as
society adopts new lifestyles. Consequently, the objective of this research was to determine the phytochemical components and
antibacterial activity of Solanum sisymbriifolium, a traditional medicinal plant used in Ethiopia to treat arthritis. Phytochemical
analyses were performed on the leaf extracts to identify the bioactive constituents. The results of this study indicated that the
plant contains carbohydrates, phenols, flavonoids, alkaloids, proteins, steroids, saponins, and terpenoids. Tannins and
anthraquinones were absent. Alkaloids and terpenoids’presence in the leaves of these plants is a potential bioactive for
bacterial inhibitors. At optimal conditions (62
°
C, 72 hr, and 1 mm particle size), the maximum extraction yield is 38:538:5±
1:15%. Crystals obtained from aqueous methanol extracts were subjected to FT-IR, and the compound spectrum showed a
characteristic absorption band for the N-H group at 3500 cm
-1
and 1700 cm
-1
for the C = O group, and the medium intensity
at 1236 cm
-1
indicates a C-O stretching. Sharp absorption at 707 cm
-1
is inductive for = C-H bending. According to agar disc
diffusion tests, plant extracts of 50 mg/mL produced 14.04 mm growth inhibition zones of Bacillus subtilis. Phytochemical and
antibacterial studies of Solanum sisymbriifolium indicated that the plant is a source of highly valued compounds for the
preparation of medications.
1. Introduction
As reported by the WHO, a medicinal plant is a bioactive
plant that can be used for remedial purposes or that is a pre-
cursor to the production of chemical and pharmaceutical
products [1]. Since ancient times, plants have long been
the main source of medicine for about 80% of the population
[2, 3]. In literature such as Vedas and the Bible, the wide
spread use of herbal medicines and health care products
obtained from commonly used traditional herbs and plants
is linked to the popularity of herbal medicine and its medical
properties [4]. From earliest times itself, plants were used for
treatment of disease without knowledge about the com-
pounds present and their mode of action. Over the centuries,
societies around the world have developed their own tradi-
tion to make sense of medicinal plants and their uses. For
example, more than 70% of Ethiopia population uses tradi-
tional medicine, and more than 95% of medicines are made
of plants [5, 6].
The wide spread use of herbal medicines and health care
preparations is obtained from commonly used traditional
plants. It has been raised due to the occurrence of natural
products with medicinal properties. Not only this, but also,
(i) they have practical experience and positive beliefs about
traditional medicine. (ii) They have a limited ability to
acquire and afford current healthcare services [6, 7].
However, traditional healers and the indigenous com-
munity believe that medicinal plants must be kept secret if
Hindawi
Advanced Gut & Microbiome Research
Volume 2023, Article ID 5525606, 7 pages
https://doi.org/10.1155/2023/5525606

they are to be effective. If healers wish to share their wisdom,
they usually choose one curious and wiser family member to
whom they impart it verbally. These beliefs and practises
have been the reason for the fast disappearance of medicinal
plants before the scientific community reached. The use of
plant extracts and phytochemicals, both with known antimi-
crobial properties, can be of great significance in therapeutic
treatments [8]. Numerous researches have been carried out
recently in various nations to demonstrate its effectiveness.
A lot of plants have been employed for their antibacterial
properties, which are the result of compounds produced
during the plant’s secondary metabolism. These products
are known by their active substances, for example, the alka-
loids, tannin, saponin, and anthraquinone compounds are a
significant increase in the study of medicinal plants as a rem-
edy for various forms of diseases and disorders [9].
In this sense, medicinal plants with historically recog-
nised bioactive ingredients offer promising prospects for
future research and drug development [10].
Solanum sisymbriifolium is an important flowering plant
species with multiple uses. In many Ethiopian communities,
it is one of the most widely used species for traditional
medicinal treatment. It has scarlet fruits and yellow flowers.
It can be distributed in wild near homes, overgrazed areas,
wastelands, and roadside areas. In the present study,
phytochemical screening and antibacterial activities studies
are presented on the crude leaf extract of Solanum
sisymbriifolium.
2. Materials and Methods
2.1. Collection and Preparation of Plant Materials. The leaves
of Solanum sisymbriifolium were collected from the
Endabagerima area, at points of latitude 14
°
09′49.2″N and
longitude 38
°
56′19.3″E Adwa, Tigray, Ethiopia. Botanical
identification of the plant was done by Mr. Abadi Gebreye-
sus Hndya at Mekelle University-Botany Laboratory. The
experiment was conducted in the Department of Health
Sciences Laboratory of Mekelle University and the Adigirat
Drug Factory. The leaves were collected from the Solanum
sisymbriifolium plant from the herbal garden. The leaves
were thoroughly rinsed with tap water and then with deio-
nised water. The water-rinsed dried leaves were air dried
for 3 days in stainless-steel sieve and finally put in to oven
at 37
°
C for 2 days. The moisture content of sample was
analysis using moisture analyser with 5 hr intervals. The
dried sample (5% moisture content) was pulverised into
powder with a vibrating mill and passed through 1, 1.5,
and 2 mm sieve sizes to remove fabric particles and stored
in refrigerator.
2.2. Extraction of Plant Materials. The different particle sizes
(1, 1.5, and 2 mm) with a total of 25 g of Solanum sisymbrii-
folium powder were weighted using an expert pro electrical
balance and inserted into the Soxhlet extractor. 80% v/v of
methanol was used as extraction solvent. The experimental
design was setup with Design-Expert version 11.1.0 software.
Response surface methodology (RMS)–Box-Behnken (BB)
(Table 1) and crude extracts were filtered using Whatman
No. 1 filter paper. Upon observing the homogeneity of the
data, contents, the methanol-based Soxhlet extraction, and
filtration procedures were repeated three times to increase
the extraction and filtration efficiency. Using the developed
regression model, the process factors were optimised to pro-
duce the best percentage of extracted yield. The % yield is
calculated according to Abdisa and Kenea [11]. Based on
the above analysis, the extraction temperature (56-68
°
C),
extraction time (48-72 h), and particle size (1–2 mm) were
fed to the software, and the experimental design layouts
are given in Table 1.
2.3. Box-Behnken Design for Optimisation. The response sur-
face methodology–(RMS–) Box-Behnken (BB) design has
been applied to optimise extraction parameters with respect
to high yield. This study focuses the effects of three variables
such as temperature (56–68
°
C), particle size (1–2 mm), and
time (48–72 hr) on the crude yield. Each variable was con-
sidered at three levels. Table 1 shows the ranges and levels
of each factor. The response surface methodology–(RMS–)
Box-Behnken (BB) design matrix consisting of 17 run trials
involving three variables, each variable at three levels, was
obtained using Design-Expert software. Therefore, 17 exper-
iments with different combinations of temperature, particle
size, and time were conducted according to the BB, and
the response was recorded [12].
2.4. Phytochemical and FTIR Analysis. Standard methods
described by Harborne [13] were used to perform the qual-
itative phytochemical screening of plant extracts such as
Table 1: Experimental results (mean ± SD,n=3)ofSolanum
sisymbriifolium leaf extract in % yield.
Std Run
Factor 1 Factor 2 Factor 3 Response 1
A: temperature B: time C: particle size Yield
°
CHrmm %
6 1 68 60 1 37:75 ± 0:58
16 2 62 60 1.5 35 ± 2:31
10 3 62 72 1 38:5±1:15
15 4 62 60 1.5 35 ± 0:58
11 5 62 48 2 35 ± 0:58
2 6 68 48 1.5 35:5±1:15
3 7 56 72 1.5 37 ± 0:58
7 8 56 60 2 35:25 ± 0
14 9 62 60 1.5 35 ± 1:15
4 10 68 72 1.5 38:25 ± 0:12
12 11 62 72 2 35:25 ± 0
13 12 62 60 1.5 35 ± 2:31
913 62 48 1 34:5±1:15
514 56 60 1 36 ± 2:31
815 68 60 2 35:25 ± 0
1 16 56 48 1.5 35:75 ± 0:58
17 17 62 60 1.5 35 ± 0
2 Advanced Gut & Microbiome Research

tannins, terpenoids, anthraquinones, proteins, alkaloids, phe-
nols, carbohydrates, phlobatannin, saponins, flavonoids, and
steroids [14–22]. The crude extract was accurately weighed
and consistently mixed with KBr salt using a mortar and com-
pressed into a thin pellet. The pellet was analysed using a
Shimadzu FTIR spectroscope from 4500 to 400 cm
-1
with a
resolution of 4 cm
-1
to detect the characteristic peaks.
2.5. Antibacterial Activities of the Extract. The antimicrobial
activity of the crude extract under optimal conditions was
tested against Bacillus subtilis. The strain was obtained from
Mekelle University’s College of Veterinary Medicine’s Veter-
inary Microbiology Laboratory. The bacteria colonies were
inoculated in liquid nutrient broth and cultured at 37
°
Cin
the evening and agitated at 200 rpm. Then, each broth
culture was adjusted to fit the McFarland half-turbidity
standard to obtain around 1×10
8CFU/mL [23]. Likewise,
Mueller-Hinton media were prepared according to the pro-
cedures given by its manufacturer as growth media for the
agar disc diffusion assay [24–26]. To ensure that the
Mueller-Hinton agar medium is not polluted, plates with
the medium were developed and maintained in evening.
Similarly, 6 mm filter paper discs were prepared from sterile
Whatman No. 1 filter paper using a paper punch. M-H
media plates were distributed using plate spread technology
using 100 microlitres of 12 hr bacteria culture and dried for a
X: Actual
Y: Predicted
Predicted vs. Actual
34
35
36
37
38
39
34 35 36 37 38 39
Design-Expert® Soware
Yield
34.5 38.5
Color points by value of
Yield:
Figure 1: Graph of measured vs. predicted responses.
48
54
60
66
72
56
59
62
65
68
34
35
36
37
38
39
Yield (%)
A: Temperature (°C)
B: Time (hr)
Design-Expert® Soware
Factor coding: Actual
Yield (%)
Design points above predicted value
34.5 38.5
Actual factor
C: Particle size = 1.5
Design points below predicted value
X1 = A: Temperature
X2 = B: Time
Figure 2: Three-dimensional response surface plots for extract yield as a function of temperature and extraction time.
3Advanced Gut & Microbiome Research

minute before surface moisture was extracted. The filter
paper disc was infused with a concentration of 100 mg/
mL curd extract. The disc was then placed on the agar
plate using a stick and distributed for an hour at normal
temperature. The plates were then stored in the labora-
tory for 2 hr at room temperature. Finally, the plates were
incubated at 37
°
C for 24 hr [27]. After incubation, plates
are observed, and the diameter of the inhibition area is
measured using digital electronic measuring devices. Bac-
terial cultures with an inhibitory area greater than or
equal to 7 mm in diameter were considered resistant to
extracts [2].
56 59 62 65 68
48
54
60
66
72 Yield (%)
X: A: Temperature (°C)
Y: B: Time (hr)
35
36
37
38
5
Design-Expert® Soware
Factor coding: Actual
Yield (%)
Design points
34.5 38.5
Actual factors
C: Particle size = 1.5
X1 = A: Temperature
X2 = B: Time
Figure 3: Two-dimensional response surface contour plots for extract yield as a function of temperature and extraction time.
–1.000 –0.500 0.000 0.500 1.000
34
35
36
37
38
39
A
A
B
B
C
C
Perturbation
Design-Expert® Soware
Factor coding: Actual
Yield (%)
Actual factors
A: Temperature = 62
B: Time = 60
C: Particle size = 1.5
X: Deviation from reference point (coded units)
Y: Yield (%)
Figure 4: Perturbation vs. extract yield as a function of temperature and extraction time.
4 Advanced Gut & Microbiome Research

3. Results and Discussion
All experiments in this study were performed in triplicate,
and the results were mean values. From Table 1, it was
observed that the optimum yield is 38:538:5±1:15%that
was obtained at run three.
The plot of the actual versus predicted response
(Figure 1) were observed to fit in a straight line. It is assumed
that the suggested quadratic model is appropriate and effec-
tive for the optimisation of process variables. The regression
model equation gave precise description of the experimental
data, in which all points are very similar to the line of perfect
fit. The three-dimensional response surface plots and two-
dimensional contour plots are useful tools for anticipating
the effects of two factors on the response at the same time
and for identifying the optimum values of the independent
variables for obtaining the maximum response.
Figures 2 and 3 represent the effects of temperature and
extraction time on the extraction yield while the particle size
was kept constant. The equation in terms of actual factors
(Equation (1)) can be used to create predictions about the
response.
Perturbation disturbance graph (Figure 4) indicates the
relationship between extraction temperature (A), extraction
time (B), and particle size (C) with yields (Y). Both extrac-
tion temperature (A) and extraction time (B) increased, par-
ticle size could decrease, and yield of the plant extract
increased. The extraction yield is highly affected by extrac-
tion time (B).
From Table 2, we observed that the differences in actual
value and predicted value in each run are small means that
the model is significant [28]. The differences between the
actual values and predicted values are indicators for models
that are acceptable or not. If the differences are more, the
model is invalid, whereas if the differences are very small,
the models are significant. Finally, after omitting the AB
and AC terms, the simpler quadratic regression model was
proposed. The linear effects of temperature and time are sig-
nificant and positive, which means that by increasing them,
it is possible to increase the extraction yield.
Y=35+0:34A+1:03B−0:75C+0:38AB +0:94A2
+0:69B2+0:13C2
:
ð1Þ
According to ANOVA, extraction was significantly
influenced by extraction temperature, particle size, and
extraction time variables.
The Shimadzu FTIR spectrum analysis confirmed the
presence of 3500 cm
-1
N-H starching functional groups.
2980.75 cm
-1
indicated C-H stretching with the medium
appearance of alkane compound. Symmetric extension of
HC (CH
2
) was found at absorption of 2860 cm
-1
, the H-C
aldehyde absorption band 2750 cm
-1
. 3500 cm
-1
indicates
primary stretching of the amine medium of H-C with sin-
gle-bond region. 2250 cm
-1
is C-C medial alkyne acetylenic
with triple bond region [28]. 1750 cm
-1
is C = O starch ester
with strong double bond region. 700 cm
-1
is cis-C-H out-of-
plane bend alkene with strong fingerprint’s region.
986.11 cm
-1
is for C=C twisting [29]. The phytochemical
analysis of the crude extract (Table 3) confirmed the pres-
ence of proteins, phenols, saponins, flavonoids, alkaloids,
steroids, phlobatannins, carbohydrates, and terpenoids.
Tannins and anthraquinone were absent. The presence of
phenolics in plants indicates that plants are sources of anti-
microbial agents [22]. Flavonoids and phenolics showed that
Table 2: Overall report of actual versus predicted value of extract yield.
Run
order
Actual
value
Predicted
value
Residual
value Leverage
Internally
studentized
residual
Externally
studentized
residuals
Cook’s
distance
Influence on
fitted value
Standard
order
1 37.75 37.59 0.1562 0.750 2.500 7.071
(1)
1.875
(2)
12.247
(2)
6
2 35.00 35.00 0.0000 0.200 0.000 0.000 0.000 0.000 16
3 38.50 38.53 -0.0313 0.750 -0.500 -0.471 0.075 -0.816 10
4 35.00 35.00 0.0000 0.200 0.000 0.000 0.000 0.000 15
5 35.00 34.97 0.0313 0.750 0.500 0.471 0.075 0.816 11
6 35.50 35.56 -0.0625 0.750 -1.000 -1.000 0.300 -1.732 2
7 37.00 36.94 0.0625 0.750 1.000 1.000 0.300 1.732 3
8 35.25 35.41 -0.1562 0.750 -2.500 -7.071
(1)
1.875
(2)
12.247
(2)
7
9 35.00 35.00 0.0000 0.200 0.000 0.000 0.000 0.000 14
10 38.25 38.37 -0.1250 0.750 -2.000 -2.828 1.200
(2)
-4.899
(2)
4
11 35.25 35.16 0.0937 0.750 1.500 1.686 0.675 2.920
(2)
12
12 35.00 35.00 0.0000 0.200 0.000 0.000 0.000 0.000 13
13 34.50 34.59 -0.0937 0.750 -1.500 -1.686 0.675 -2.920
(2)
9
14 36.00 36.03 -0.0312 0.750 -0.500 -0.471 0.075 -0.816 5
15 35.25 35.22 0.0312 0.750 0.500 0.471 0.075 0.816 8
16 35.75 35.62 0.1250 0.750 2.000 2.828 1.200
(2)
4.899
(2)
1
17 35.00 35.00 0.0000 0.200 0.000 0.000 0.000 0.000 17
5Advanced Gut & Microbiome Research

the plant has potent antioxidant activity or free radical scav-
engers [30]. Alkaloids in medicinal plants have been
reported as an important antimicrobial and analgesic drug
alkaloids and terpenoids which have a potential antibacterial
activity in medicine [31]. Saponins are known to produce an
inhibitory effect on inflammation [32]. Saponins are essen-
tial in the treatment of cough and in the controlling of
soreness of the upper respirational region. Furthermore,
plant-based saponins serve primarily as tonics for the heart
and have been documented to prevent diabetics and inhibit
fungi growth [33]. Terpenoids have been found to be useful
in the prevention of antimicrobial, antifungal, antiparasitic,
antiallergenic, and anti-inflammatory properties. Phlobatan-
nins have been reported to possess astringent properties [18].
The antimicrobial activity of the crude extract under
optimal conditions was tested against standard strains of B.
subtilis bacteria. According to the diffusion tests of the agar
disc, 50 mg/ml of crude extract under optimal plant condi-
tions produced bacterial growth inhibition zones of
14.04 mm in diameter, which were found to be more effec-
tive. Bacterial cultures with an inhibition zone of ≥7mm in
diameter were remarkable antimicrobial activities [2].
4. Conclusions
The optimal extraction yield achieved under optimal condi-
tions of 62
°
C, 72 h, and 1 mm particle size is 38:538:5±1:15
%. The findings of this study demonstrate that plants con-
tain significant bioactive substances, such as terpenoids
and alkaloids, which prevent the growth of bacterial strains
[31]. The results suggest that the leaf of Solanum sisymbriifo-
lium contains secondary metabolites, indicating that the
plant is the source of highly valued compounds for drug
preparation.
Data Availability
The data that was used to support the study are available in
the manuscript.
Conflicts of Interest
The authors declare that they have no conflicts of interest
that could have appeared to influence the work reported in
this paper.
Acknowledgments
The authors are grateful to Mekelle University and Adigrat
Drug Factory for the funding and infrastructural facilities
provided to carry out the work, respectively. This study
was funded by Mekelle University under the small scale
research grant.
References
[1] K. Karunamoorthi, M. Phil, K. Jegajeevanram, J. Vijayalakshmi,
and E. Mengistie, “Traditional medicinal plants: a source of
phytotherapeutic modality in resource-constrained health care
settings,”Journal of Evidence-Based Complementary & Alterna-
tive Medicine, vol. 18, no. 1, pp. 67–74, 2013.
[2] D. B. Sbhatu and H. B. Abraha, “Preliminary antimicrobial
profile of solanum incanum L.: a common medicinal plant,”
Evidence-Based Complementary and Alternative Medicine,
vol. 2020, Article ID 3647065, 6 pages, 2020.
[3] G. Gebreyohannes, A. Nyerere, C. Bii, and D. B. Sbhatu,
“Investigation of antioxidant and antimicrobial activities of
different extracts of auricularia and termitomyces species of
mushrooms,”The Scientific World Journal, vol. 2019, Article
ID 7357048, 10 pages, 2019.
[4] L. Hoareau and E. J. DaSilva, “Medicinal plants: a re-emerging
health aid,”Electronic Journal of Biotechnology, vol. 2, no. 2,
1999.
[5] N. Tuasha, B. Petros, and Z. Asfaw, “Medicinal plants used by
traditional healers to treat malignancies and other human ail-
ments in Dalle District, Sidama Zone, Ethiopia,”Journal of
Ethnobiology and Ethnomedicine, vol. 14, no. 1, pp. 1–21, 2018.
[6] B. N. Tefera and Y. Kim, “Ethnobotanical study of medicinal
plants in the Hawassa Zuria District, Sidama Zone, Southern
Ethiopia,”Journal of Ethnobiology and Ethnomedicine,
vol. 15, no. 1, pp. 1–21, 2019.
[7] B. Abera, “Medicinal plants used in traditional medicine by
Oromo people, Ghimbi District, Southwest Ethiopia,”Journal
of Ethnobiology and Ethnomedicine, vol. 10, no. 1, pp. 1–15,
2014.
[8] M. Thenmozhi and R. Sivaraj, “International journal of
pharma and bio sciences phytochemical analysis and antimi-
crobial activity of Polyalthia longifolia materials and
methods,”International Journal of Pharma and Bio Sciences,
vol. 1, no. 3, pp. 1–7, 2010.
[9] F. Othman, M. S. Sadeghian, F. Ebrahimi, and M. Heydari, “A
study on sedimentation in Sefidroud Dam by using depth eval-
uation and comparing the results with USBR and FAO
methods,”International Proceedings of Chemical, Biological
and Environmental Engineering, vol. 51, no. 9, p. 6, 2013.
[10] R. Article, “Preparation of medicinal plants: basic extraction
and fractionation procedures for experimental purposes,”
Journal of Pharmacy & Bioallied Sciences, vol. 12, no. 1,
pp. 1–10, 2020.
Table 3: Qualitative phytochemical screening results.
No. Test Result
1 Carbohydrates +
2 Alkaloids (Mayer test) +
3 Terpenoids +
4 Steroids +
5 Flavonoids +
6 Tannins -
7 Saponin +
8 Phenol +
9 Anthraquinone -
10 Phlobatannins +
11 Protein +
+ = present; - = absent.
6 Advanced Gut & Microbiome Research

[11] Z. Abdisa and F. Kenea, “Phytochemical screening, antibacte-
rial and antioxidant activity studies on the crude root extract of
Clematis hirsuta,”Cogent Chemistry, vol. 6, no. 1, 2020.
[12] S. N. H. Azmi, B. M. H. Al-Jassasi, H. M. S. Al-Sawafi,S.H.G.
Al-Shukaili, N. Rahman, and M. Nasir, “Optimization for
synthesis of silver nanoparticles through response surface
methodology using leaf extract of Boswellia sacra and its appli-
cation in antimicrobial activity,”Environmental Monitoring
and Assessment, vol. 193, no. 8, p. 497, 2021.
[13] J. B. Harborne, “Metode Fitokimia : Penuntun Cara Modern
Menganalisis Tumbuhan,”1987, http://perpustakaan.stifar.ac
.id/index.php?p=show_detail&id=448&keywords=harborne.
[14] K. S. Banu and L. Cathrine, “General Techniques Involved in
Phytochemical Analysis,”International Journal of Advanced
research in Chemical Science, vol. 2, no. 4, pp. 25–32, 2015.
[15] S. Paul, K. Mukherjee, A. De, A. Bhattacharyya, and S. Basu,
“Process optimization for preparation of an antibacterial agent
using a combined extract of tulsi and marigold leaves,”IOSR
Journal of Environmental Science, Toxicology and Food Tech-
nology, vol. 11, no. 5, pp. 31–35, 2017.
[16] N. Jaradat, F. Hussen, and A. Al Ali, “Preliminary phytochem-
ical screening, quantitative estimation of total flavonoids, total
phenols and antioxidant activity of Ephedra alata,”Journal of
Materials and Environmental Science, vol. 6, no. 6, pp. 1771–
1778, 2015.
[17] S. Bhaisare Manmohan and S. G. Kunjalwar, “Phytochemical
screening of E. chaetaria, (Roem. & Schult), Cyperaceae
Bhandara District of Maharashtra,”International Journal Life
Sciences, vol. 3, no. 1, pp. 91–95, 2015.
[18] M. Yadav, S. Chatterji, S. K. Gupta, and G. Watal, “Preliminary
phytochemical screening of six medicinal plants used in
traditional medicine,”International Journal of Pharmacy and
Pharmaceutical Science, vol. 6, no. 5, 2014.
[19] A. A. Abdalla, M. I. Mustafa, and A. M. Makhawi, “Phyto-
chemical screening and antimicrobial activities studies of Aca-
cia nilotica fruit cover,”bioRxiv, 2020.
[20] M. Merghem, S. Dahamna, and S. Khennouf, “Polyphenols
contents and antioxidant activity of extracts from leaves and
flowers of Thymelaea hirsuta,”Journal of Drug Delivery and
Therapeutics, vol. 10, no. 4, pp. 108–111, 2020.
[21] T. Thilagavathi, R. Arvindganth, D. Vidhya, and R. Dhivya,
“Preliminary phytochemical screening of different solvent
mediated medicinal plant extracts evaluated research,”Inter-
national Research Journal of Pharmacy, vol. 10, no. 4,
pp. 246–248, 2015.
[22] A. Doss, M. Sc, and M. Phil, “Preliminary phytochemical
screening of some Indian medicinal plants,”Ancient Science
of Life, vol. 29, no. 2, pp. 12–16, 2009.
[23] S. Kothari, V. Mishra, S. Bharat, and S. D. Tonpay, “Antimi-
crobial activity and phytochemical screening of serial extracts
from leaves of Aegle marmelos (Linn.),”Acta Poloniae Phar-
maceutica - Drug Research, vol. 68, no. 5, pp. 687–692, 2011.
[24] E. Karimi, H. Z. E. Jaafar, and S. Ahmad, “Phytochemical anal-
ysis and antimicrobial activities of methanolic extracts of leaf,
stem and root from different varieties of labisa pumila benth,”
Molecules, vol. 16, no. 6, pp. 4438–4450, 2011.
[25] M. D. Makut, S. D. Gyar, G. R. I. Pennap, and P. Anthony,
“Phytochemical screening and antimicrobial activity of the
ethanolic and methanolic extracts of the leaf and bark of Khaya
senegalensis,”African Journal of Biotechnology, vol. 7, no. 9,
pp. 1216–1219, 2008.
[26] C. N. Durga, M. Lakshmi, B. Mahitha, T. Madhavi, and N. J.
Sushma, “Phytochemical screening and FTIR analysis of Cli-
toria Ternatea leaves,”International Journal of Scientific and
Engineering Research, vol. 6, no. 2, pp. 287–290, 2015.
[27] A. P. S. A. Da Silva, L. C. Nascimento da Silva, C. S. Martins da
Fonseca et al., “Antimicrobial activity and phytochemical anal-
ysis of organic extracts from Cleome spinosa Jaqc,”Frontiers
in Microbiology, vol. 7, pp. 1–10, 2016.
[28] I. T. Karabegović, S. S. Stojičević, D. T. Veličković,N.Č. Niko-
lić, and M. L. Lazić,“Optimization of microwave-assisted
extraction and characterization of phenolic compounds in
cherry laurel (Prunus laurocerasus) leaves,”Separation and
Purification Technology, vol. 120, pp. 429–436, 2013.
[29] A. Bayu, D. Nandiyanto, R. Oktiani, and R. Ragadhita, “How
to read and interpret ftir spectroscope of organic material,”
Indonesian Journal of Science & Technology, vol. 4, no. 1,
pp. 97–118, 2019.
[30] G. A. Ayoola, H. A. Coker, S. A. Adesegun et al., “Phytochem-
ical screening and antioxidant activities of some selected
medicinal plants used for malaria therapy in Southwestern
Nigeria,”Tropical Journal of Pharmaceutical Research, vol. 7,
no. 3, pp. 1019–1024, 2008.
[31] A. M. Karande, H. V. Kamble, V. H. Kumbhar, S. R. Kane, C. S.
Magdum, and S. R. Kane, “Preliminary phytochemical screen-
ing of Glochidion ellipticum,”European Journal of Experimen-
tal Biology, vol. 6, no. 4, pp. 41–45, 2016.
[32] O. A. Aiyegoro and A. I. Okoh, “Preliminary phytochemical
screening and in vitro antioxidant activities of the aqueous
extract of Helichrysum longifolium DC,”BMC Complemen-
tary and Alternative Medicine, vol. 10, no. 1, 2010.
[33] A. C. Marcus, O. S. Edori, and M. C. Maduagu, “Phytochemi-
cal and anti-microbial screening of Phyllantus fratenus and
Taraxacuim officinale leaves,”Biochemistry & Analytical Bio-
chemistry, vol. 8, pp. 1009–2161, 2019.
7Advanced Gut & Microbiome Research