ISSN 1519-6984 (Print)

ISSN 1678-4375 (Online)

THE INTERNATIONAL JOURNAL ON NEOTROPICAL BIOLOGY

THE INTERNATIONAL JOURNAL ON GLOBAL BIODIVERSITY AND ENVIRONMENT

Original Article

Phytochemistry, allelopathy and anticancer potentiality of

Withania somnifera (L.) Dunal (Solanaceae)
Fitoquímica, alelopatia e potencialidade anticancerígena de Withania somnifera (L.)
Dunal (Solanaceae)

H. A. Ahmeda,b*  and S. M. El-Darierb 

King Faisal University, College of Science, Biological Sciences Department, Al-Ahsa, Saudi Arabia
a

b
Alexandria University, Faculty of Science, Botany and Microbiology Department, Alexandria, Egypt

Abstract
Withania somnifera is a wild plant that shows great activity and safety against several human diseases. The current
research explored the plant’s chemical composition and allelopathic effects on Rumex dentatus (recipient plant).
Moreover, anticancer activity is also tested against four types of human cancer cell lines. Chemical analysis of W.
somnifera showed a high percentage of saponins and tannins, while glycosides, alkaloids, and flavonoids occurred
in the second order. Results of the allelopathic experiments revealed significant inhibition of the R. dentatus
plumule and radicle lengths as well as their relative dry weights. In addition, significant reductions in some primary
metabolites of R. dentatus, like non-reducing and total sugar as well as soluble proteins, were determined. Cytotoxic
potentiality of W. somnifera was also proved against four different cancer lines, namely; human hepatocellular
carcinoma cell line (HepG2), human non-small cell lung cancer cell line (A549), human breast cancer cell line
(MCF7), and colon cancer cell line (CaCo2) with IC50 value of about 38, 19, 27, and 24 𝜇g/ml, respectively.
Keywords: Withania somnifera; Rumex dentatus; germination; growth; metabolites; cytotoxic potential.

Resumo
Withania somnifera é uma planta silvestre que apresenta grande atividade e segurança contra diversas doenças
humanas. A presente pesquisa explorou a composição química da planta e os efeitos alelopáticos em Rumex dentatus
(planta receptora). Além disso, a atividade anticancerígena também é testada contra quatro tipos diferentes de
linhagens de células cancerígenas humanas. A análise química de W. somnifera mostrou alta porcentagem de
saponinas e taninos, enquanto glicosídeos, alcaloides, e flavonoides ocorreram na segunda ordem. Os resultados
dos experimentos alelopáticos revelaram uma inibição significativa dos comprimentos de plúmula e radícula
de R. dentatus, bem como seus pesos secos relativos. Além disso, foi determinada redução significativa em
alguns metabólitos primários de R. dentatus como não redutores e açúcar total, bem como proteínas solúveis. A
potencialidade citotóxica de W. somnifera também foi comprovada contra quatro diferentes linhas de câncer, a
saber: linha celular de carcinoma hepatocelular humano (HepG2), linha celular de câncer de pulmão de células
não pequenas humanas (A549), linha celular de câncer de mama humano (MCF7) e linha celular de câncer de
cólon (CaCo2) com valor de IC50 de cerca de 38, 19, 27 e 24 𝜇g/ml, respectivamente.
Palavras-chave: Withania somnifera; Rumex dentatus; germinação; crescimento; metabólitos; potencial citotóxico.

1. Introduction

Wild plants produce many biochemicals in the of carbon steel in HCl solution (Fouda et al., 2021).
environment that may positively or negatively influence The species was distributed mainly in the neglected
other organisms and communities. Allelopathy is a branch areas next to buildings and garden flora of cities. Roots
of chemical ecology that studies the effects of such contain flavonoids, alkaloids, steroid, polyphenols and
biochemical on organisms’ germination, productivity, many bioactive functional constituents (Kumar et al.,
yield, and metabolic changes (Cheng and Cheng, 2015). 2015). The chemistry of the plant has been extensively
Withania somnifera (Ashwagandha) is one of the studied, leading to the isolation and characterization of
valuable perennial plant species with wide therapeutic several groups of chemical constituents, which are of great
applications in traditional and modern system of medicine biological and pharmacological interests. Aerial parts,
(Datta et al., 2011) as well as a friendly corrosion inhibitor roots and berries of this species considered as important

*e-mail: haahmed@kfu.edu.sa
Received: May 15, 2022 – Accepted: October 4, 2022
This is an Open Access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use,
distribution, and reproduction in any medium, provided the original work is properly cited.

Brazilian Journal of Biology, 2024, vol. 84, e263815 | https://doi.org/10.1590/1519-6984.263815 1/8

Ahmed, H.A. and El-Darier, S.M.

sources of some secondary metabolites like 12 or more flavonoids and phenolic compounds. One gram of dry root
from alkaloids, 40 withanolides and several sitoindosides sample was weighed and soaked in ethanol (ethanolic
(Dhar et al., 2015; Mirjalili et al., 2009). Besides these, the extract) and water (aqueous extract). The sample was
plant comprises a number of other secondary metabolites, saturated and allowed to stay overnight for 24 hours.
including flavonols glycosides, sterols, phenolics, glycosides, After overnight incubation with continuous shaking the
starch, reducing sugar, and a variety of amino acids sample extract was filtered with Whatman filter paper
including aspartic acid, proline, tyrosine, alanine, glycine, then the filtrate was centrifuged at 10,000 to 25,000 rpm
glutamic acid, cysteine, tryptophan and high amount of
for 10 minutes. After that, the supernatant was used for
iron (Gupta and Rana, 2007).
phytochemical screening (Soni et al., 2012). The supernatant
There is always an increase in research interest in
was kept in a refrigerator until use.
identifying bioactive molecules that have a role in treating
cancer or protecting against it. In particular, molecules
extracted or isolated from plants are most attractive because 2.3. Collection and preparation of recipient seeds
of their safety, cost-effectiveness and feasibility of oral Healthy uniform seeds of R. dentatus were collected
administration. Nutraceuticals have played an important from wheat and broad bean fields in El-Bihara province in
role in the overall well-being of humans for many years, with June 2019. The seeds were scratched with sandpaper and
or without rigorous evidence backing their health claims soaked in distilled water for 1 h, then surface sterilized
(Abid et al., 2024). Traditional medicine systems worldwide with 70% ethanol for 2 min and rinsed with double-distilled
have utilized plants for millennia that have medicinal water several times for complete removal of the sterilant.
properties, providing an opportunity for modern-day
researchers to assess their efficacies against ailments such
2.4. Allelopathy experiment
as cancer (Mendonça et al., 2020; Palliyaguru et al., 2016).
Withania somnifera is a plant that has been demonstrated To perform the germination bioassay test, the surface
to have anti-tumorigenic properties in experimental models sterilized seeds were placed evenly in sterilized glass
(Dutta et al., 2019). While scientific research on the plant Petri dishes (9 mm). Each Petri dish contained 10 seeds.
has exploded in the past decade, much regarding the Then equal volumes (5 ml) of varying concentrations of
mode of action and molecular targets involved remains the aqueous extract were introduced into each Petri dish.
unknown. In the present study, the phytochemistry, Similar volumes of double distilled water were used as
allelopathy and anticancer potentiality were performed control. All the Petri dishes were incubated in the dark
to evaluate W. somnifera as a potent herbicide against at room temperature (24-26°C). The allelopathic behavior
Rumex dentatus L., (Polygonaceae) and antitumor agent was evaluated by recording the number of germinated
in plants and humans. The two species are invasive and seeds, plumule (PL) and radicle (RL) lengths using a
waste-land weeds. millimeter ruler and plumule (PDW) and radicle (RDW)
dry weights for 7 days after sowing. The vigor Index (VI)
was calculated according to Vashisth and Nagarajan (2010)
2. Materials and Methods as follows (Equation 1):

2.1. Collection and extract preparation

= Vigor Index (VI ) Germination percentage x mean of seedling length ( plumule + radicle ) (1)
Withania somnifera was collected from Alexandria, Egypt,
in August 2020. S.M. El-Darier identified the plant, and a Germination and inhibition percentages (GP & IP) of
voucher specimen has been deposited in the Herbarium of radicle growth were calculated by the following formulae
Botany and Microbiology Department, Faculty of Science, (Equation 2):
Alexandria University, Alexandria, Egypt.
Fresh W. somnifera roots of good quality were obtained
= Germination percentage Number of germinated seeds / Total number of seeds × 100
in a single lot from herbal garden, washed under running Inhibition ( % ) of radicle
= growth ( X – Y ) / X × 100
(2)
water to remove adhering foreign particles, weighed fresh,
and dried at low temperature (45-55°C), then ground
where, X= Control mean radicle length and Y= Treated
using an electric grinder. The powder obtained was sieved
mean radicle length.
twice to remove the coarse particles and stored in air-tight
containers until further analysis. The yield of the dry matter
2.5. Determination of primary metabolites
was calculated as about 69%. Ten grams of root powder
were soaked in ethanol in a 1:10 w/v ratio for 24 hours. Seven-day-old R. dentatus seedlings were used for the
The extract was filtered and alcohol was evaporated using extraction and determination of carbohydrates as reducing
rotary evaporator apparatus under low temperature. sugar (Hernández-López et al., 2020), soluble protein
The residue was used later in vitro cytotoxic assay. (Singh et al., 2015), and total amino acids (Lee et al., 2013).

2.2. Phytochemical screening 2.6. Potential cytotoxicity of W. somnifera

The root extract of W. somnifera was analyzed for Ethanolic extract was tested using the method of
the presence of alkaloids, glycosides, saponins, steroids, (Skehan et al., 1990) by using Sulforhodamine B (SRB) Assay

2/8 Brazilian Journal of Biology, 2024, vol. 84, e263815

 Biological activity of Withania somnifera

at the National Cancer Institute, Cairo, Egypt by serial sub glycosides, alkaloids and flavonoids occurred in the second
culturing. HepG2, A549, MCF7, and CaCo2 were tested. order (++). Fats and fixed oils and volatile oils were not
1- Cells were plated in a 96- multiwall plate (104 cells/ well) detected in the two types of extracts, and carbohydrates
for 24 hours before treatment with the compound (s) in ethanol extract only. Total phenolics constituents
to allow attachment of the cell to the wall of the plate. identified by RP-HPLC analysis at λ = 280 nm (Table 2) and
2- Different concentrations of the compound under test at λ = 330 nm of the ethanolic extracts (Table 3) exerted
(0, 1, 2.5, and 10 µg) were added to the cell monolayer high values of about 8.03% (at λ = 280 nm) and 13.95% (at
triplicate wells and were prepared for each dose. λ = 330 nm). The results showed the absence of catechin
3- Monolayer cells were incubated with the compound (s) and rutin at λ = 280 and 330 nm respectively.
for 48 hours at 37˚C and in an atmosphere of 5% CO2.
4- After 48 hours at 37˚C, Cells were fixed, washed, and
stained with Sulforhodamine B. Table 1. Qualitative screening of the two plant extracts.
5- The excess stain was washed with acetic acid, and the
Ethanolic Aqueous
attached stain was recovered with Tris-EDTA buffer. Test
extract extract
6- Color intensity was measured using an ELISA reader.
7- The relation between surviving fraction and drug Alkaloids ++ +
concentration is plotted to get the survival curve of Carbohydrates - +
each cancer cell line after the specified treatment.
Fats and fixed oil - -
Surviving fractions of cells throughout drug exposure
were characterized graphically by IC 50 values (drug Flavonoids ++ +
concentration that yields 50% fewer cells than the drug- Glycosides ++ +
free control). The IC50 values were estimated by linear Proteins and amino acids + +
least-squares regression of the growth values versus the
Saponins + +++
logarithm of the extract concentration; only concentrations
that yielded growth values between 10 and 90% were Tannins + +++
used in the calculation. National Cancer Institute (NCI, Volatile oil - -
USA) recommended that 30 µg/ml as the upper IC50 limit
(-) The active compound under investigation was not found;
is considered promising for purification of a crude extract (+) Weak to moderate amounts of the active compound under
(Mothana et al., 2009). investigation; (++, +++) high amounts of active compound under
investigation.
Predominantly, a bar (P) parallel to the x-axis and
intersecting the point 50% on the y-axis was constructed.
In the next step, a bar was plotted parallel to the y-axis,
Table 2. Phenolics identified by RP-HPLC* analysis (at λ = 280 nm)
starting from the point of intersection of P with the dose-
of the ethanolic extracts of Withania somnifera.
response plot. The IC50 could then be directly determined
at the point of intersection with the x-axis. Identified
Retention time Relative area (%)
constituent
2.7. Growth Inhibition Percentage (GIP) 6.81 Pyrogallol 0.13
Growth inhibition percentage (GIP) was calculated 6.92 Gallic acid 0.04
according to the general equations of Mosmann (1983)
8.235 Protocatechuic 0.15
(Equation 3):
8.444 Catechin -
GIP
= 100 − ( Treated survival cells / control cells ) * 100  (3)
  8.593 Chlorogenic acid 0.48

8.950 Catechol 0.13

2.8. Statistical analysis
Data concerning the effect of different concentrations 10.040 Caffeic acid 0.22

of W. somnifera aqueous extract on some germination 11.073 Vanillic acid 0.40

and growth criteria as well as some metabolites were 11.620 Ferulic acid 0.36
subjected to standard analysis of variance (ANOVA) (Zar,
12.466 Salicylic acid 0.69
1984) using the COSTAT 2.00 statistical analysis software
manufactured by Cohort software. 12.943 Ellagic acid 1.20

13.127 Benzoic acid 1.04

13.789 Coumaric acid 0.55

3. Results
14.980 Cinnamic acid 0.84

3.1. Phytochemical screening 18.657 Chrysin 1.80

The phytochemical screenings of W. somnifera are Total identified

8.03
presented in Table 1. Data showed that the plant contains constituents
appreciable amounts (+++) of saponins and tannins while *Reverse-phase high performance liquid chromatography.

Brazilian Journal of Biology, 2024, vol. 84, e263815 3/8

Ahmed, H.A. and El-Darier, S.M.

3.2. Allelopathic potentiality same trend and the effect was concentration dependent.
Results in Figure 1 showed significant inhibition in Notably, the inhibition percentage increased significantly
seed germination percentages of R. dentatus treated with as the extract concentration increased.
W. somnifera extracts at the concentrations inclined from The remarkable reduction in the growth of the tested
control to 8% concentration level. Similarly, seedling growth weed species was accompanied by a significant reduction
of the weed species was highly affected with donor species in reducing, non-reducing and total sugar content as well
extracts showing a reduction in the plumule and radicle as soluble proteins. Contrarily, free amino acid content
lengths as well as total seedling length and their relative increased significantly as extract concentration increased
dry weights. Consequently, the vigor index (VI) attained the (Figure 2).

3.3. Anti-proliferative activity of W. somnifera ethanolic

Table 3. Phenolics identified by RP-HPLC analysis (at λ = 330 nm) extract
of the ethanolic extracts of Withania somnifera. In vitro cytotoxicity, W. somnifera was tested on four
human cancer cell lines. Dose response curves constructed
Identified
Retention time Relative area (%) for the SRB method between 19-92 µg/ml ranges. Results
constituent
in Figure 3 showed that the incubation of W. somnifera
3.83 Quercetin 0.04 extract with HepG2 and A549 significantly inhibited cell
11.78 Rosmarinic acid 2.65 proliferation with IC50 values of about 38 and 19 𝜇g/ml,
respectively. Likewise, apoptosis in MCF7 was enhanced
12.06 Hesperidin 1.13
by W. somnifera with an IC50 value of 27𝜇g/ml, whereas
12.43 Rutin - the potential cytotoxic activity of W. somnifera against
13.267 Quercitrin 3.82 CaCo2 attained IC50= 24 𝜇g/ml.

14.576 Naringenin 1.00

3.4. Growth inhibition percentage
14.952 Hesperitin 1.20
The samples of W. somnifera root ethanolic extract
15.147 Kampferol 1.11 were verified on four cell lines (HepG2, A549, MCF7 and
16.167 Apigenin 1.00 CACo2). Growth inhibition percentage (GIP) is illustrated
and statistically represented in Figure 4. The radar
Total identified
11.95 presentation method displayed values relative to the
constituents
center point. The proliferation of HepG2 was significantly

Figure 1. Response of germination and some growth criteria of Rumex dentatus seedlings to variation in extract concentration (%) of Withania
somnifera 7 days after sowing. Different letters on bars indicate a significant difference at P < 0.05 according to one-way ANOVA test.

4/8 Brazilian Journal of Biology, 2024, vol. 84, e263815

 Biological activity of Withania somnifera

Figure 2. Response of some metabolic products (mg/g.d.w.) in Rumex dentatus seedlings to variation in extract concentration (%) of Withania
somnifera 7 days after sowing. Different letters on bars indicate a significant difference at P < 0.05 according to one-way ANOVA test.

inhibited by 80.2% suppression. Likewise, the proliferation

of MCF7 and CACo2 was significantly inhibited by 79 and
68.9%, respectively, while the proliferation of A459 was
inhibited by 59.5%.

4. Discussion

Medicinal plants are a rich source of bioactive

phytochemicals or bionutrients. A proper understanding
of phytochemical screening for plants is essential for
drug discovery, developing novel therapeutic agents and
for interpreting the allelopathic effects of several plant
species (Anwar et al., 2020; Egbuna et al., 2018). Studies
carried out during the past 2– 3 decades have shown
that phytochemicals have an important role in ecosystem
management and in preventing chronic diseases like cancer
(Saxena et al., 2013). The chemical analyses infer the
effective role that plant extracts play and the mechanism by
which they work, whether on the ecological or therapeutic
levels (Altemimi et al., 2017; Kamal et al., 2022).
The present study explored that in W. somnifera root
extract, saponins and tannins attained the first order
followed by glycosides, alkaloids and flavonoids. Total
phenolics constituents exerted considerable high values.
The interrelationship between these constituents and the
apparent interaction of some germination and growth
factors in the recipient R. dentatus plant was significant.
Seed germination percentages, and seedling growth
(plumule and radicle lengths, total seedling length and
their relative dry weights, vigorous index) were reduced,
accompanied by a reduction in some metabolites (reducing,
Figure 3. In vitro cytotoxicity of W. somnifera on four human
non-reducing and total sugar content as well as soluble
cancer cell lines.
proteins). In contrarily, free amino acid content increased
significantly as extract concentration increased. Li et al.
(2010) stated that phenolic allelochemicals had been declines in crop yield due to soil sickness. For example,
observed in natural and managed ecosystems, where they in agroecosystems, allelochemicals have detrimental
cause many ecological and economic problems, such as effects on the growth of associated and next-season

Brazilian Journal of Biology, 2024, vol. 84, e263815 5/8

Ahmed, H.A. and El-Darier, S.M.

important among various anticancer medicinal plants.

It is essential to further screen and investigate different
formulations for anticancer therapy in vitro and in vivo in
combination with established chemotherapy.

Acknowledgements

The authors acknowledge the Deanship of Scientific

Research (DSR), Vice Presidency for Graduate Studies and
Scientific Research, King Faisal University, Saudi Arabia,
for the financial support under the research grant (Project
No. GRANT1825).

Figure 4. In vitro cytotoxicity of Withania somnifera ethanolic

References
extracts against four cell lines HepG2: human liver hepatocellular
carcinoma cell line, A549: human non-small cell lung carcinoma ABID, F., SALEEM, M., LEGHARI, T., RAFI, I., MAQBOOL, T., FATIMA,
cell line, MCF7: human breast carcinoma cell line and CACO2: colon F., ARSHAD, A.M., KHURSHID, S., NAZ, S., HADI, F., TAHIR, M.,
cancer cell line in the SRB assay using radar method. AKHTAR, S., YASIR, S., MOBASHAR, A. and ASHRAF, M., 2024.
Evaluation of in vitro anticancer potential of pharmacological
ethanolic plant extracts Acacia modesta and Opuntia monocantha
crops (Ridenour and Callaway, 2001). The current study against liver cancer cells. Brazilian Journal of Biology = Revista
illustrated that W. somnifera root aqueous extract (WSRAE) Brasileira de Biologia, vol. 84, pp. e252526. http://dx.doi.
affects the germination percentage (GP) of R. dentatus due org/10.1590/1519-6984.252526.
to phytotoxic allelopathic interactions. Additionally, the ALTEMIMI, A., LAKHSSASSI, N., BAHARLOUEI, A., WATSON, D.G.
obtained data established that inhibition in germination and LIGHTFOOT, D.A., 2017. Phytochemicals: extraction,
percentage (IP) of R. dentatus was significantly affected isolation, and identification of bioactive compounds from plant
due to the allelopathic action of the WSRAE. Ordinarily, extracts. Plants, vol. 6, no. 4, p. 42. http://dx.doi.org/10.3390/
IP increased with the increase of WSRAE concentration plants6040042. PMid:28937585.
and reached the maximum value of 51.25% at 8% WSRAE ANWAR, T., QURESHI, H., PARVEEN, N., BASHIR, R., QAISAR, U., MUNAZIR,
concentration. These results agreed with Shteliyana et al. M., YASMIN, S., BASIT, Z., MAHMOOD, R.T., NAYYAR, B.G., KHAN,
(2012) on seeds of soybean, peas and vetch in response to S., KHAN, S.A., QURESHI, M.M. and WALI, M., 2020. Evaluation of
bioherbicidal potential of Carica papaya leaves. Brazilian Journal of
higher concentrations of the Johnson grass roots extract
Biology = Revista Brasileira de Biologia, vol. 80, no. 3, pp. 565-573.
and with that recorded by Raoof and Siddiqui (2012).
http://dx.doi.org/10.1590/1519-6984.216359. PMid:31644650.
Many therapeutics with low side effects are of natural
origin (Newman and Cragg, 2007; Sabir et al., 2021; CHENG, F. and CHENG, Z., 2015. Research progress on the use of
plant allelopathy in agriculture and the physiological and
Santana et al., 2021). In this respect, plants are extensively
ecological mechanisms of allelopathy. Frontiers in Plant Science,
exploited as a potential source for active components with
vol. 6, p. 1020. http://dx.doi.org/10.3389/fpls.2015.01020.
high antitumor activity. Due to the side effects of chemical PMid:26635845.
drugs, the use of herbs for medicinal purposes is expanding
DATTA, A.K., DAS, A., BHATTACHARYA, A., MUKHERJEE, S. and
in the clinical field (Yuan et al., 2016). Continuing oxidative
GHOSH, B.K., 2011. An overview on Withania somnifera (l.)
damage to lipids, proteins, DNA, and other molecules
dunal – the ‘Indian ginseng. Medicinal and Aromatic Plant
may contribute to cancer development (Neri et al., 2021; Science and Biotechnology, vol. 5, pp. 1-15.
Thanan et al., 2015).
DHAR, N., RAZDAN, S., RANA, S., BHAT, W.W., VISHWAKARMA, R.
Approximately 60% of all drugs now undergoing clinical
and LATTOO, S.K., 2015. A decade of molecular understanding
trials for the multiplicity of cancers are either natural of withanolide biosynthesis and in vitro studies in Withania
products or compounds derived from natural products somnifera (L.) Dunal: prospects and perspectives for pathway
(Réthy, 2007). Earlier studies indicated that only the engineering. Frontiers in Plant Science, vol. 6, p. 1031. http://
root extract of W. Somnifera is a potential source of new dx.doi.org/10.3389/fpls.2015.01031. PMid:26640469.
molecules that can curtail cancer growth (Dredge et al., DREDGE, K., DALGLEISH, A.G. and MARRIOTT, J.B., 2003. Angiogenesis
2003). W. Somnifera leaves have also been shown to inhibit inhibitors in cancer therapy. Current Opinion in Investigational
the growth of human cancer cell lines comparared to that Drugs, vol. 4, no. 6, pp. 667-674. PMid:12901224.
produced by Adriamycin (Yadav et al., 2010). In this study, DUTTA, R., KHALIL, R., GREEN, R., MOHAPATRA, S. and MOHAPATRA,
growth inhibitory activity of root of W. somnifera was S., 2019. Withania somnifera (Ashwagandha) and withaferin
investigated against four cell lines representing four different A: potential in integrative oncology. International Journal
tissues, HepG2, A549, MCF7, and CaCo2. Current results clearly of Molecular Sciences, vol. 20, no. 21, p. 5310. http://dx.doi.
indicated that the cytotoxic effect of W. somnifera beyond org/10.3390/ijms20215310. PMid:31731424.
the four human cancer cell lines resulted in promising EGBUNA, C., IFEMEJE, J.C., KRYEZIU, T.I., MUKHERJEE, M., SHAH, H.,
starting sources for further investigation. The study supports RAO, G.M.N., GIDO, L.J.F.J. and TIJJANI, H., 2018. Introduction to
that W. somnifera ethanol extract was highly cytotoxic to phytochemistry. In: C. EGBUNA, J.C. IFEMEJE, S.C. UDEDI and S.
the studied human cell lines. In conclusion, this herb is KUMAR, eds. Phytochemistry: fundamentals, modern techniques,

6/8 Brazilian Journal of Biology, 2024, vol. 84, e263815

 Biological activity of Withania somnifera

and applications. New York: Academic Press, vol. 1, pp. 3-35. 83, pp. e236649. http://dx.doi.org/10.1590/1519-6984.236649.
http://dx.doi.org/10.1201/9780429426223-1. PMid:34669789.
FOUDA, A.S., AHMED, A.M., EL-DARIER, S.M. and IBRAHIM, I.M., 2021. NEWMAN, D.J. and CRAGG, G.M., 2007. Natural products as sources
Withania somnifera extract as a friendly corrosion inhibitor of of new drugs over the last 25 years. Journal of Natural Products,
carbon steel in HCl solution. International Journal of Corrosion vol. 70, no. 3, pp. 461-477. http://dx.doi.org/10.1021/np068054v.
and Scale Inhibition, vol. 10, no. 1, pp. 245-261. PMid:17309302.
GUPTA, G.L. and RANA, A.C., 20 07. Withania somnifera PALLIYAGURU, D.L., SINGH, S.V. and KENSLER, T.W., 2016. Withania
(Ashwagandha): a review. Pharmacognosy Reviews, vol. 1, pp. somnifera: from prevention to treatment of cancer. Molecular
129-136. Nutrition & Food Research, vol. 60, no. 6, pp. 1342-1353. http://
dx.doi.org/10.1002/mnfr.201500756. PMid:26718910.
HERNÁNDEZ-LÓPEZ, A., FÉLIX, D.A.S., SIERRA, Z.Z., BRAVO, I.G.,
DINKOVA, T.D. and AVILA-ALEJANDRE, A.X., 2020. Quantification RAOOF, K.M.A. and SIDDIQUI, M.B., 2012. Evaluation of allelopathic
of reducing sugars based on the qualitative technique of impact of aqueous extract of root and aerial root of Tinospora
Benedict. American Chemical Society Omega, vol. 5, no. 50, pp. cordifolia (willd.) miers on some weed plants. Analele Universitatii
32403-32410. http://dx.doi.org/10.1021/acsomega.0c04467. din Oradea. Fascicula Biologie, vol. 1, pp. 29-34.
PMid:33376877. RÉTHY, B., 2007. Antitumor effect of plant extracts and their
KAMAL, Y., KHAN, T., HAQ, I., ZAHRA, S.S., ASIM, M.H., SHAHZADI, constituents on cancer cell lines. Szeged: Department of
I., MANNAN, A. and FATIMA, N., 2022. Phytochemical and Pharmacodynamics and Biopharmacy, Faculty of Pharmacy,
biological attributes of Bauhinia variegata L. (Caesalpiniaceae). University of Szeged. Ph.D. thesis.
Brazilian Journal of Biology = Revista Brasileira de Biologia, vol. RIDENOUR, W.M. and CALLAWAY, R.M., 2001. The relative importance
82, pp. e257990. http://dx.doi.org/10.1590/1519-6984.257990. of allelopathy in interference: the effects of an invasive weed
PMid:35170677. on a native bunchgrass. Oecologia, vol. 126, no. 3, pp. 444-450.
http://dx.doi.org/10.1007/s004420000533. PMid:28547460.
KUMAR, V., DEY, A., HADIMANI, M.B., MARCOVIĆ, T. and EMERALD,
M., 2015. Chemistry and pharmacology of Withania somnifera: SABIR, S.M., ZEB, A., MAHMOOD, M., ABBAS, S.R., AHMAD, Z. and
an update. Cell Medicine, vol. 5, no. 1, pp. 1.1-1.13. http://dx.doi. IQBAL, N., 2021. Phytochemical analysis and biological activities
org/10.5667/tang.2014.0030. of ethanolic extract of Curcuma longa rhizome. Brazilian Journal of
Biology = Revista Brasileira de Biologia, vol. 81, no. 3, pp. 737-740.
LEE, L.S., CHOI, J.H., SON, N., KIM, S.H., PARK, J.D., JANG, D.J., JEONG,
http://dx.doi.org/10.1590/1519-6984.230628. PMid:32965334.
Y. and KIM, H.J., 2013. Metabolomic analysis of the effect of
shade treatment on the nutritional and sensory qualities of green SANTANA, C.B., SOUZA, J.G.L., TOLEDO, A.G., ALVES, L.F.A., ALVES,
tea. Journal of Agricultural and Food Chemistry, vol. 61, no. 2, pp. D.S., CORRÊA, J.M. and PINTO, F.G.S., 2021. Antimicrobial and
332-338. http://dx.doi.org/10.1021/jf304161y. PMid:23256790. insecticidal effects of essential oil and plant extracts of Myrcia
oblongata DC in pathogenic bacteria and Alphitobius diaperinus.
LI, Z.H., WANG, Q., RUAN, X., PAN, C. and JIANG, D., 2010. Phenolics Brazilian Journal of Biology = Revista Brasileira de Biologia, vol.
and plant allelopathy. Molecules, vol. 15, no. 12, pp. 8933-8952. 82, pp. e233425. http://dx.doi.org/10.1590/1519-6984.233425.
http://dx.doi.org/10.3390/molecules15128933. PMid:21139532. PMid:34105655.
MENDONÇA, L.A.B.M., MATIAS, R., ZANELLA, D.F.P., PORTO, K.R.A., SAXENA, M., SAXENA, J., NEMA, R., SINGH, D. and GUPTA, A., 2013.
GUILHERMINO, J.F., MOREIRA, D.L., ROEL, A.R., POTT, A. and A phytochemistry of medicinal plants. Journal of Pharmacognosy
CARVALHO, C.M.E., 2020. Toxicity and phytochemistry of eight and Phytochemistry, vol. 1, no. 6, pp. 168-182.
species used in the traditional medicine of sul-mato-grossense,
SHTELIYANA, K., IRENA, G., ANGEL, H. and ANNA, I., 2012. Allelopathic
Brazil. Brazilian Journal of Biology = Revista Brasileira de Biologia,
effect of aqueous extract from root systems of Johnson grass
vol. 80, no. 3, pp. 574-581. http://dx.doi.org/10.1590/1519-
on seed germination and initial development of soybean, pea
6984.216406. PMid:31644651.
and vetch. Ratarstvo i Povrtarstvo, vol. 49, no. 3, pp. 250-256.
MIRJALILI, M.H., MOYANO, E., BONFILL, M., CUSIDO, R.M. and PALAZÓN, http://dx.doi.org/10.5937/ratpov49-1200.
J., 2009. Steroidal lactones from Withania somnifera, an ancient
SINGH, S., KUMAR, P., TRIVEDI, A. and KOSHAL, A., 2015. Seed protein
plant for novel medicine. Molecules, vol. 14, no. 7, pp. 2373-2393.
profiling through electrophoresis in mungbean [V.radiata (L.)
http://dx.doi.org/10.3390/molecules14072373. PMid:19633611.
Wilczek]. Journal of Plant Development, vol. 7, no. 4, pp. 337-340.
MOSMANN, T., 1983. Rapid colorimetric assay for cellular growth SKEHAN, P., STORENG, R., SCUDIERO, D., MONKS, A., MCMAHON, J.,
and survival: application to proliferation and cytotoxicity VISTICA, D., WARREN, J.T., BOKESCH, H., KENNEY, S. and BOYD,
assays. Journal of Immunological Methods, vol. 65, no. 1-2, M.R., 1990. New colorimetric cytotoxicity assay for anticancer-
pp. 55-63. http://dx.doi.org/10.1016/0022-1759(83)90303-4. drug screening. Journal of the National Cancer Institute, vol. 82,
PMid:6606682. no. 13, pp. 1107-1112. http://dx.doi.org/10.1093/jnci/82.13.1107.
MOTHANA, R.A., LINDEQUIST, U., GRUENERT, R. and BEDNARSKI, PMid:2359136.
P.J., 2009. Studies of the in vitro anticancer, antimicrobial and SONI, K.A., DESAI, M., OLADUNJOYE, A., SKROBOT, F. and
antioxidant potentials of selected Yemeni medicinal plants NANNAPANENI, R., 2012. Reduction of Listeria monocytogenes
from the island Soqotra. BMC Complementary and Alternative in queso fresco cheese by a combination of listericidal and
Medicine, vol. 9, no. 1, p. 7. http://dx.doi.org/10.1186/1472- listeriostatic GRAS antimicrobials. International Journal of
6882-9-7. PMid:19320966. Food Microbiology, vol. 155, no. 1-2, pp. 82-88. http://dx.doi.
NERI, T.S., SILVA, K.W.L., MAIOR, L.P.S., OLIVEIRA-SILVA, S.K., org/10.1016/j.ijfoodmicro.2012.01.010. PMid:22305889.
AZEVEDO, P.V.M., GOMES, D.C.S., SOUZA, M.A., PAVÃO, J.M.S.J., THANAN, R., OIKAWA, S., HIRAKU, Y., OHNISHI, S., MA, N., PINLAOR, S.,
COSTA, J.G., CUNHA, A.L., FERREIRA-JÚNIOR, G.C., MATOS- YONGVANIT, P., KAWANISHI, S. and MURATA, M., 2015. Oxidative
ROCHA, T.J., SANTOS, A.F. and FONSECA, S.A., 2021. Phytochemical stress and its significant roles in neurodegenerative diseases
characterization, antioxidant potential and antibacterial activity and cancer. International Journal of Molecular Sciences, vol. 16,
of the Croton argyrophylloides Muell. Arg. (Euphorbiaceae). no. 1, pp. 193-217. http://dx.doi.org/10.3390/ijms16010193.
Brazilian Journal of Biology = Revista Brasileira de Biologia, vol. PMid:25547488.

Brazilian Journal of Biology, 2024, vol. 84, e263815 7/8

Ahmed, H.A. and El-Darier, S.M.

VASHISTH, A. and NAGARAJAN, S., 2010. Effect on germination Journal of Pharmaceutical Sciences, vol. 72, no. 5, pp. 659-663.
and early growth characteristics in sunflower (Helianthus http://dx.doi.org/10.4103/0250-474X.78543. PMid:21695006.
annuus) seeds exposed to static magnetic field. Journal of YUAN, H., MA, Q., YE, L. and PIAO, G., 2016. The traditional medicine
Plant Physiology, vol. 167, no. 2, pp. 149-156. http://dx.doi. and modern medicine from natural products. Molecules, vol.
org/10.1016/j.jplph.2009.08.011. PMid:19783321. 21, no. 5, p. 559. http://dx.doi.org/10.3390/molecules21050559.
YADAV, B., BAJAJ, A., SAXENA, M. and SAXENA, A.K., 2010. In vitro PMid:27136524.
anticancer activity of the root, stem and leaves of Withania ZAR, J.H., 1984. Biostatistical analysis. 2nd ed. Englewood Cliffs:
Somnifera against various human cancer cell lines. Indian Prentice-Hall, Inc., 718 p.

8/8 Brazilian Journal of Biology, 2024, vol. 84, e263815