Chemical Composition, Biological Activity, and
Health-Promoting Effects of Withania somnifera for Pharma-
Food Industry Applications
Javad Shariﬁ-Rad ,
Seyed Abdulmajid Ayatollahi ,
Karl R. Matthews,
Hari Prasad Devkota ,
Miquel Martorell ,
Haﬁz Ansar Rasul Suleria ,
Mohammed M. Alshehri ,
alia Cruz-Martins ,
and William C. Cho
Phytochemistry Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Facultad de Ciencias de la Salud, Universidad Arturo Prat, Avda. Arturo Prat 2120, Iquique 1110939, Chile
Department of Pharmacognosy and Biotechnology, School of Pharmacy, Shahid Beheshti University of Medical Sciences,
Department of Medicinal Chemistry, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Institute of Soil Science and Plant Cultivation–State Research Institute, Czartoryskich 8, Puławy 24-100, Poland
Molecular and Applied Mycology and Plant Pathology Laboratory, Department of Botany, University of Calcutta,
Kolkata 700019, India
Department of Botany, Fakir Chand College, Diamond Harbour, West Bengal 743331, India
Department of Food Science, Rutgers University, New Brunswick, New Jersey, USA
Gazi University, Faculty of Pharmacy, Department of Pharmacognosy, Ankara 06330, Turkey
Graduate School of Pharmaceutical Sciences, Kumamoto University, 5-1 Oe-honmachi, Kumamoto 862-0973, Japan
Department Food Engineering, Faculty of Chemical and Metallurgical Engineering, Istanbul Technical University, Maslak,
Istanbul 34469, Turkey
Bioactive Research & Innovation Food Manufacturing Industry Trade Ltd. Co., Maslak, Istanbul 34469, Turkey
Department of Nutrition and Dietetics, Faculty of Pharmacy, University of Concepci´on, Concepci´on 4070386, Chile
Centre for Healthy Living, University of Concepci´
on 4070386, Chile
Department of Agriculture and Food Systems, e University of Melbourne, Melbourne 3010, Australia
Pharmaceutical Care Department, Ministry of National Guard-Health Aﬀairs, Riyadh, Saudi Arabia
Department of Veterinary Sciences and Animal Husbandry, Amrita School of Agricultural Sciences,
Amrita Vishwa Vidyapeetham University, Coimbatore 642109, India
Chemical and Biochemical Processing Division, ICAR–Central Institute for Research on Cotton Technology,
Mumbai 400019, India
Faculty of Medicine, University of Porto, Porto, Portugal
Institute for Research and Innovation in Health (i3S), University of Porto, Porto, Portugal
Institute of Research and Advanced Training in Health Sciences and Technologies (CESPU), Rua Central de Gandra, 1317,
Gandra 4585-116, Portugal
TOXRUN–Toxicology Research Unit, University Institute of Health Sciences, CESPU, CRL, 4585-116 Gandra, Portugal
Department of Clinical Oncology, Queen Elizabeth Hospital, Kowloon, Hong Kong
Correspondence should be addressed to Javad Shariﬁ-Rad; javad.shariﬁrad@gmail.com, Hari Prasad Devkota; devkotah@
gpo.kumamoto-u.ac.jp, Miquel Martorell; email@example.com, Nat´alia Cruz-Martins; firstname.lastname@example.org, and William
C. Cho; email@example.com
Received 9 July 2021; Revised 19 October 2021; Accepted 10 December 2021; Published 29 December 2021
Academic Editor: Sobhy El-Sohaimy
Journal of Food Quality
Volume 2021, Article ID 8985179, 14 pages
Copyright ©2021 Javad Shariﬁ-Rad et al. is 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
e Withania genus comes from the Solanaceae family and includes around 23 species, spread over some areas of the Med-
iterranean, Asia, and East Africa. Widely used in traditional medicine for thousands of years, these plants are rich in secondary
metabolites, with special emphasis on steroidal lactones, named withanolides which are used as ingredients in numerous
formulations for a plethora of diseases, such as asthma, diabetes, arthritis, impotence, amnesia, hypertension, anxiety, stress,
cancer, neurodegenerative, and cardiovascular diseases, and many others. Among them, Withania somnifera (L.) Dunal is the
most widely addressed species from a pharmacological and agroindustrial point of view. In this sense, this review provides an
overview of the folk uses, phytochemical composition, and biological activity, such as antioxidant, antimicrobial, anti-in-
ﬂammatory, and cytotoxic activity of W. somnifera, although more recently other species have also been increasingly investigated.
In addition, their health-promoting eﬀects, i.e., antistress, anxiolytic, adaptogenic, antirheumatoid arthritis, chemoprotective, and
cardiorespiratory-enhancing abilities, along with safety and adverse eﬀects are also discussed.
e genus Withania (Solanaceae) includes 23 species ,
mostly occurring in North Africa, Canary Islands, Southern
Europe, and Asia (Figure 1) [2–7]. Of the known species,
there are two of huge economic importance that are also
mostly grown due to their wide applicability in natural
medicine , namely, Withania somnifera (L.) Dunal and
Withania coagulans (Stocks) Dunal. Both species are grown
mainly in subtropical regions of India. However,
W. somnifera even presents a greater economic signiﬁcance
[9, 10]. In Morocco and Algeria, Withania adpressa Cors. is
also found as an endemic species , although both the
morphological form and phytochemical composition of
such plants undergo polymorphisms, conditioned by its
occurrence in a given geographical area .
Although various Withania spp. have been used in
traditional medicine for the management of diﬀerent pa-
thologies , W. somnifera and W. coagulans are the most
widely recognized species not only for their economic value
but also for their therapeutic potential, and they are largely
commercialized and cultivated in Afghanistan, Iran, India,
and Pakistan [13–20]. In this sense, this review aims to
provide an overview of the botanical features, traditional
uses, phytochemical composition, biological activities, and
health-promoting eﬀects observed in preclinical and clinical
studies of W. somnifera, along with updated data on its safety
and adverse eﬀects.
2. Botanical Features
Plants under the Withania genus are evergreen with heights
ranging from 0.5 to 2.0 m, present grasses, bush suburbs,
branched or unbranched [21, 22]. e ﬂowers are green or
yellow, little pedicelled or pentameric umbels, sessile to
subsessile, and hermaphrodites. ey have simple leaves,
petiolate, ovate, alternate, or in unequal pairs with a sharp
apex. Fruits are berry of 6 mm in diameter, with orange-red
color when mature, globous, and enclosed in the green calyx.
Seeds are compressed, small, ﬂat, yellow, reniform, reticulate
to smooth, and very light [2, 23–28].
3. Traditional Uses
From a folk medicinal point of view, W. somnifera, known as
“winter cherry,” is the most important species belonging to
the Withania genus, and that evidences the most renowned
therapeutic abilities. is plant has been used in Indian
medicine for a long time, and its roots are used in more than
200 formulations [2, 29, 30].
W. somnifera (called Ashwagandha, Indian ginseng) is the
best-known species, widely used in natural medicine as it
helps in many diﬀerent ailments, namely, in boosting the
immune and hematopoietic system, has an anti-inﬂammatory
activity that helps in skin diseases and osteoarthritis, and also
has antiaging eﬀects. In addition, it is also used in hypo-
thyroidism, cardiovascular diseases, diabetes, depression, and
chronic stress [31, 32]. More recently, several clinical trials
have also conﬁrmed their therapeutic uses, namely, in the
treatment of anxiety, insomnia, and Parkinson’s disease .
In Ayurveda, W. somnifera is used for over 3000 years  and
is considered to have excellent rejuvenating abilities, while it
prolongs life and has strong aphrodisiac eﬀects. Indeed, this
plant is traditionally used in India to promote youthful vigor,
strength, endurance, and health [20, 33], so that such re-
storative properties have led to W. somnifera roots being
called Indian ginseng. W. somnifera may also be useful to treat
various central nervous system (CNS) disorders, such as
epilepsy, stress, and neurodegenerative conditions, like Par-
kinson’s disease (PD), Alzheimer’s disease (AD), and even
cerebral ischemia. Ethnobotanically, it can be used as a
hallucinogenic agent .
With the rising number of literature available, it has also
been indicated that such species may also exert cytotoxic
eﬀects, opening the possibility of its use in oncological
therapies. According to Verma and Kumar , the che-
mopreventive properties of W. somnifera make it a poten-
tially useful adjunct for patients undergoing radiation and
chemotherapy. W. somnifera stimulates the immune system
by stimulating the production of T lymphocytes and mac-
rophages [35, 36], while Ziauddin et al.  stated a general
increase in the number of white blood cells after adminis-
tration of a root extract. W. somnifera application has also
2Journal of Food Quality
been shown to be able to reduce the number of skin lesions
relative to the control group and showed inhibition of cancer
cell growth in breast, lung, and colon cancer, which, apart
from its cytotoxic abilities, is linked to their excellent an-
tioxidant eﬀects [38, 39]. Other authors, namely, Panda and
Kar  and Andallu and Radhika , also stated an in-
crease in T4 thyroid hormone concentration following
W. somnifera root powder application, so that its use may be
helpful in controlling the levels of hormones in diseases
linked to hypothyroidism. Some authors have also indicated
that W. somnifera root may be used for preventing car-
diovascular disease, such as atherosclerosis [40–42]. For
instance, in a human trial, a signiﬁcant decrease in blood
glucose and cholesterol levels to the extent of 10% and 12%,
respectively, was observed when compared to the group that
received the conventional oral drug for type 2 diabetes
(Daonil). ese therapeutic eﬀects could be due to one or
more active principles in the roots of the plant. e hy-
poglycemic eﬀect of W. somnifera root could be speciﬁcally
attributed to its ability to enhance serum insulin levels and/
or the antioxidant activities of catalase, superoxide dis-
mutase, and glutathione peroxidase [40–42].
Chemical analysis of diﬀerent plant parts of W. somnifera
has aﬀorded numerous compounds belonging to various
chemical classes. e biologically active chemical
constituents of W. somnifera are alkaloids (isopelletierine,
anaferine), steroidal lactones (withanolides, withaferins),
saponins containing an additional acyl group (sitoindoside
VII and VIII), and withanolides with glucose at carbon 27
(sitoindoside XI and X). Among them, withanolides (ste-
roidal lactones) have been used in an increasing number of
drug formulations, given their promissory therapeutic
Despite being widely reported by a plethora of studies,
Table 1 and Figure 2 present some of the most important
withanolides isolated from Withania spp., considering its
abundance and bioactive eﬀects and representative structures,
respectively. Misra et al.  reported withanolide A, with-
anolide B, 27-hydroxy withanolide B, withanolide D, with-
aferin A, 16β-acetoxy-6α, 7α-epoxy-5α–hydroxy-1-oxowitha-
2, 17 (20), 24-trienolide, 5, 7α-epoxy-6α, 20α–dihydroxy-1-
oxowitha-2, 24- dienolide along with common steroids,
β-sitosterol and sitosterol, and their glucosides in
W. somnifera. Matsuda et al.  isolated 7 new withanolide
glycosides from W. somnifera roots, named withanoside I to
VII, among which class VI is more abundant. Similarly,
Bessalle and Lavie  isolated two chlorinated withanolides,
namely, withanolide C and 4-deoxyphysalolactone from dried
leaves of W. somnifera (Table 1).
ere have been also reports on other constituents from
plants of the Withania genus, namely, fatty acids and volatile
compounds. Misra et al.  have reported new ergosterol
and 1, 4-dioxane derivatives along with various fatty acids
Figure 1: Red spots indicate the geographical distribution of Withania spp.
Journal of Food Quality 3
(octacosane, oleic and stearic fatty acids), steroids, and ole-
anolic acid from W. somnifera roots. For example, Rautela
et al.  studied the constituents of both ethanol and
methanol extracts of W. somnifera leaves and roots and
analyzed components by gas chromatography-mass spec-
trometry (GC-MS). Various compounds, including with-
anolide B, rosifoliol, and phytol, were reported . Gulati
et al.  studied the chemical composition of various ex-
tracts from W. somnifera roots of diﬀerent genotypes and
stated several metals in its composition, along with diﬀerent
concentrations of total sugars, alkaloids, and tannins. Bhatia
et al. , studying the eﬀect of chemotype variations in the
chemical composition of W. somnifera fruits using GC-MS
and nuclear magnetic resonance (NMR) spectroscopy, stated
clear variations in metabolites contents in diﬀerent
5. Biological Activities
Given the wide range of Withania species applications in
Ayurvedic medicine for multiple aims, an increasing
number of studies have progressively addressed their bio-
logical eﬀects (Figure 3). Furthermore, with the populari-
zation, the use of this plant as a food supplement in the
market is also increasing. Indeed, both extracts and com-
pounds isolated from the Withania species exhibit excellent
Table 1: List of selected withanolides and other compounds identiﬁed from Withania somnifera (L.) Dunal.
parts Compounds References
Withanolide A, withanolide B, 27-hydroxy withanolide B, withanolide D, withaferin A, 16β-acetoxy-6α, 7α-epoxy-
5α–hydroxy-1-oxowitha-2, 17 (20), 24-trienolide, 5, 7α-epoxy-6α, 20α–dihydroxy-1-oxowitha-2, 24-dienolide 
Withanoside I, withanoside II, withanoside III, withanoside IV, withanoside V, withanoside VI, withanoside VII,
withaferin A, physagulin D, coagulin Q 
Withasilolide A, withasilolide B, withasilolide C, withasilolide D, withasilolide E, withasilolide F 
Withanolide E, withanolide F, withanolide G, withanolide H, withanolide I, withanolide J, withanolide K,
withanolide L, withanolide M 
Withanolide Q, withanolide R 
Withanolide E, withanolide F, withanolide S, withanolide P 
Withanolide T, withanolide U 
Stem bark Withasomnilide, withasomniferanolide, somniferanolide, somniferawithanolide, somniwithanolide 
Withanolide C, 4-deoxyphysalolactone 
(20R, 22R)-14α, 2OαF-dihydroxy-1-oxowitha-2, 5, 16, 24-tetraenolide 
Withaferin A 
24,25-Dihydrowithanolide A, withanolide A, withanone, withaferin A, 27-hydroxy withanone, and 17-hydroxy
withaferin A, 27-deoxy-16-en-withaferin A, 2, 3-dihydro-3β-hydroxywithanone, 2,3-dihydro withanone-3β-O-
Fruits 24,25-Dihydrowithanolide VI, withanoside IV, withanoside V, withanoside VI, withanamide A, withanamide B,
withanamide C, withanamide D, withanamide E, withanamide F, withanamide G, withanamide H, withanamide I 
withanolide A withanolide B withanolide D withaferin A
OH OH OH OH
Figure 2: Chemical structures of some withanolide derivatives isolated from Withania somnifera.
4Journal of Food Quality
biological activities, including antioxidant, antimicrobial,
anti-inﬂammatory, and chemopreventive abilities, as
assessed by both in vitro and in vivo studies. Concerning its
in vitro biological eﬀects, studies performed so far generally
focused on their antioxidant activity and total phenolic
content (spectrophotometric and/or chromatographic
analyses) [61–68] and antimicrobial eﬀects (disc diﬀusion
assay and/or minimum inhibitory concentration (MIC))
[65, 69–81]. In addition to in vitro studies, there has been a
signiﬁcant number of in vivo studies addressing the anti-
proliferative, cytotoxic, and anti-inﬂammatory eﬀects of
W. somnifera extracts in animal models .
Figure 3: Major biological activities of Withania somnifera. Anticancer eﬀects: W. somnifera exerts anticancer eﬀects via multiple pathways,
including nuclear factor (NFK-beta) and signal transducer and activator of transcription 3 (STAT3) signaling, PI3K (phosphoinositide 3-
kinase)/AKT (a serine-threonine protein kinase) and mitogen-activated protein kinase (MAPK) signaling, angiogenesis inhibition, oxi-
dative stress induction, and p53 signaling. Melanoma cells were destroyed by withaferin A via ROS-mediated apoptosis. is process
activated the mitochondrial pathway, resulting in the downregulation of Bcl-2, translocation of Bax to the mitochondrial membrane, release
of cytochrome c into the cytosol, abolition of transmembrane potential, and concomitant activation of caspases 9 and 3, resulting in the
downregulation of proapoptotic protein, poly (ADP-Ribose) polymerase-1 (Parp-1) and DNA fragmentation. Neuroprotection: Withania
somnifera reduced blood glucose, tissue lipid peroxidation (LPO), and glutathione (GSH) levels while increasing the activities of antioxidant
enzymes such as glutathione peroxidase (GPx), glutathione reductase (GR), glutathione-S-transferase (GST), superoxide dismutase (SOD),
and catalase (CAT). is demonstrates W. somnifera’s signiﬁcant free radical scavenging activity, as well as its ability to improve non-
enzymatic and enzymatic antioxidants. W. somnifera root extract and withanolide A protected isolated hippocampus cells against hypobaric
hypoxia-induced memory loss and neurodegeneration in vitro by stimulating the glutathione production pathway and decreasing glu-
tathione (GSH) concentration. Furthermore, in cortical neurons treated with amyloid beta peptide, Withanolide A promoted both axonal
and dendritic change as well as synaptic repair. Antidiabetic eﬀects: W. somnifera leaf and root extracts showed antidiabetic activity by
normalizing glucose uptake in skeletal myotubes and adipocytes in a dose-dependent manner. Furthermore, it considerably attenuated
levels of urine and blood glucose, glucose 6-phosphatase, and tissue glycogen levels through nonenzymatic and enzymatic antioxidant
mechanisms. Antimicrobial eﬀects: the antimicrobial eﬀect of Withania somnifera is attributed by inhibiting acid formation, acid tolerance,
bioﬁlm formation, spore germination, and hyphae growth, which in turn is mediated through gene silencing, immunopotentiation and
cytotoxicity. Cardioprotective and antistress eﬀects: the cardioprotective and cardiotropic properties of W. somnifera are demonstrated via
nuclear factor erythroid 2-related transcription factor (Nrf)-2 and by activating phase II detoxiﬁcation enzymes and abrogating apoptosis.
Moreover, it is capable of alleviating chronic stress induced reduction of T-cell population and upregulated 1 cytokines, thereby ensuring
better stress endurance in animals as well as humans. Anti-inﬂammatory and antiarthritic eﬀects: Withania somnifera alleviated in-
ﬂammation by suppressing cytokines such as interleukin- (IL-) 8 and 1, tumor necrosis factor- (TNF-) α, nitric oxide (NO), and reactive
oxygen species (ROS). Furthermore, withaferin A, one of the active ingredients of W. somnifera, inhibited the expression of cell adhesion
molecules, leukocyte adhesion and migration, IL-6 and TNF-a production, and NF-k activation (nuclear factor kappa-light-chain-enhancer
of activated B cells). Furthermore, it inhibited the phosphorylation of p38, extracellular regulated kinases (ERK 12), and c-Jun N-terminal
kinase by phorbol-12-myristate-13-acetate (PMA) (JNK).
Journal of Food Quality 5
5.1. Antioxidant Activity. e biological eﬀects, and partic-
ularly the antioxidant potential and phytochemical constitu-
ents of W. somnifera, along with the other plants of the
Withania genus, vary depending on the extraction method
. Methanol-chloroform-water (1 : 1 :1) extract of
W. somnifera roots, with the highest content of all phyto-
chemical constituents except tannins, had higher antioxidant
and reducing activities when compared to water, acetone, and
aqueous methanol (1 : 1) extracts (i,e. total antioxidant capacity
of methanol-chloroform-water (1 : 1 :1) was 83.354 ±1.828,
aqueous methanol (1 : 1) was 76.978 ±2.210, and water was
68.439 ±1.000) . Alkaloid content was found to be a leading
contributor to the overall antioxidant and reducing activities of
the extracts, closely followed by ﬂavonoids and withanolides.
Moreover, diﬀerent parts of the plant may have diﬀerent levels
of antioxidant capacity . For instance, Sumathi and Padma
 reported that the leaves and fresh and dry tubers of
W. somnifera had high contents in antioxidant compounds,
while those present in tender roots and stems were not so high.
Similar ﬁndings were also stated in other studies [63–65], with
Alam et al.  also reporting that W. somnifera presents a
good antioxidant activity, with catechin being the major
polyphenol present in the highest concentration (13.01 ±8.93
to 30.61 ±11.41 mg/g). High concentrations of polyphenols
(gallic, syringic, benzoic, p-coumaric, and vanillic acids as well
as catechin, kaempferol, and naringenin), ﬂavonoids, and
DPPH (1, 1-diphenyl-2-picrylhydrazyl) radical scavenging
activities were detected in 80% methanolic extracts of
W. somnifera fruits, roots, and leaves, ranging from
17.80 ±5.80 to 32.58 ±3.16 mg/g (dry weight), 15.49 ±1.02 to
31.58 ±5.07 mg/g, and 59.16 ±1.20 to 91.84 ±0.38 mg/g, re-
spectively . Other authors also reported that W. somnifera
root extract (0.7 and 1.4 mg/kg daily by gastric intubation
method for 20 days) improves oxidative damage due to lead
intoxication in mice by signiﬁcantly decreasing lipid perox-
idation and signiﬁcantly increasing superoxide dismutase and
catalase enzyme activities . Free radical scavenging activity
(FRSA) and metabolic proﬁle of in vitro cultivated and ﬁeld-
grown Withania somnifera roots were examined by Samir et al.
. In vitro produced roots had signiﬁcantly higher levels of
FRSA, total phenolic content (TPC), and total ﬂavonoid
content (TFC) than ﬁeld-grown samples. Furthermore, as
compared to 45-day-cultured samples, 30 day-cultured in vitro
root samples had considerably greater FRSA, TPC, and TFC.
Gas chromatography-mass spectrometry study detected a total
of 29 compounds in in vitro cultivated and ﬁeld-grown roots.
Alcohols, organic acids, purine, pyrimidine, sugars, and pu-
trescine were among the metabolites identiﬁed. Vanillic acid
was found only in in vitro cultured root samples, and it was
found in higher concentrations in 30 day-cultured in vitro root
samples than in 45 day-cultured samples. As a result, 30 day-
cultured in vitro root samples are recommended as a substitute
for ﬁeld-grown roots in the development of medicinal and
functional food products.
5.2. Anticancer, Anti-Inﬂammatory, and Cytotoxic Activity.
Regarding the anticancer and cytotoxic eﬀects of Withania
species, Samir et al.  reported that ethanol extracts of aerial
parts of W. somnifera demonstrated cytotoxic activity against
human liver (HEPG-2) and breast (MCF-7) cell lines with half-
maximal inhibitory concentration (IC
) of 8.5 μg/mL and
9.4 μg/mL for HEPG-2 and MCF-7, respectively. Cytotoxic
activity of W. somnifera extracts was found to be at the stage of
the G2/M phase and sub-G0 by arresting the cell cycle.
Similarly, Naidoo et al.  reported that W. somnifera root
extract eﬀectively regulates the levels of the inﬂammatory
cytokines while inhibiting the cancer cells’ growth. Closely
linked to the antioxidant activity, the cytotoxic activity of
W. somnifera leaf extract against hepatocellular carcinoma cell
line was also reported by Ahmed et al. . In another study, it
was observed that hydroalcoholic extract of W. somnifera root
exhibited chemopreventive activity in mice with skin cancer
 and ﬁbrosarcoma . Similarly, Padmavathi et al. 
reported that W. somnifera root exerts chemopreventive eﬀects
against forestomach and skin carcinogenesis in mice.
On the other hand, closely linked to both antioxidant
and anti-inﬂammatory eﬀects, Khadrawy et al.  reported
that W. somnifera demonstrated excellent eﬀects against
aluminum chloride (AlCl
)-induced neurotoxicity in rats.
Aluminum increased lipid peroxidation and nitric oxide
levels in the cortex, hippocampus, and striatum while
lowering glutathione levels in the hippocampus and stria-
tum. Lipid peroxidation, nitric oxide, and reduced gluta-
thione levels were not signiﬁcantly diﬀerent in rats protected
with W. somnifera extract. Furthermore, it inhibited the
increased activity of acetylcholinesterase and Na
ATPase in the cortex, hippocampus, and striatum caused by
, apart from preventing a signiﬁcant increase in tumor
necrosis factor-αinduced by AlCl
in the cortex and hip-
pocampus. ese ﬁndings imply that W. somnifera extract
can protect against aluminum neurotoxicity by acting as an
antioxidant and anti-inﬂammatory agent. Furthermore, it
helps to prevent the decline in cholinergic activity by
maintaining normal acetylcholinesterase activity. e latter
eﬀect may support the use of W. somnifera as a memory
booster. Also, Pingali et al.  reported that withaferin A of
W. somnifera can cause type II collagen expression and
increase reactive oxygen species and cyclooxygenase-2 ex-
pression in rabbit articular chondrocytes depending on dose
5.3. Cardioprotective Activity. Udayakumar et al.  sug-
gested that the ﬂavonoids and phenolics present in both root
and leaf extracts of W. somnifera can be eﬀective in reducing
the blood glucose levels in diabetic rats. It was also reported
that W. somnifera was eﬀective in decreasing hyperlipidemia
and oxidative stress in type 2 diabetic rats. When
W. somnifera was given orally to type 2 diabetic rats at
dosages 200 mg/kg and 400 mg/kg, it led to signiﬁcantly
reduced serum levels of total cholesterol, triglyceride, low-
density lipoprotein-cholesterol, and very-low-density lipo-
protein-cholesterol while high-density lipoprotein-choles-
terol levels increased signiﬁcantly when compared to the
diabetic control group . Moreover, Udayakumar et al.
 claimed that phenolic contents of the extracts of
W. somnifera leaf and root were helpful in decreasing blood
6Journal of Food Quality
glucose levels in diabetic rats. Elkady and Mohamed  also
reported that W. somnifera can be eﬀective in protecting the
occurrence of cardiotoxic eﬀects induced by c-rays in rats. A
similar ﬁnding was also reported by Hosny Mansour and
Farouk Hafez  that W. somnifera reduced hepatotoxicity
in rats exposed to c-radiation by signiﬁcantly lowering se-
rum hepatic enzymes, hepatic nitrate/nitrite, and malon-
dialdehyde levels, signiﬁcantly increasing antioxidant
activity, and signiﬁcant heme oxygenase (HO-1) induction.
HO-1 enzymes protect the cell from injury due to oxidative
and pathological stress, having a central role in cardiovas-
cular protection .
5.4. Antimicrobial Activity. e antimicrobial activity of the
Withania species is also remarkable. For example, methanol
extracts of W. somnifera roots, fruits, and leaves have been
revealed to be highly eﬀective against gram-negative bac-
teria, including Klebsiella pneumoniae, Citrobacter freundii,
Salmonella typhi, Pseudomonas aeruginosa, and Escherichia
coli, as shown by Alam et al. . Modulation of physio-
logical functions of gut microbiota is involved in the mode of
action of Withania somnifera root extracts. Similarly, the
dichloromethane and ethyl acetate extracts of aerial parts of
W. somnifera also evidenced excellent eﬀects against
Staphylococcus aureus and methicillin-resistant Staphylo-
coccus aureus by disc diﬀusion assay, as shown by Mwitari
et al.  and Hussain et al. .
e antimicrobial activity depends on the extraction
method where ethanolic and methanolic extracts of
W. somnifera root did not exhibit antibacterial activity against
K. pneumoniae and methicillin-resistant S. aureus, whereas
these microorganisms were inhibited by chloroform extracts
of stem and leaves . Moreover, the antimicrobial activities
of diﬀerent extracts of W. somnifera against diﬀerent bacteria
were reported by AbdEislam et al. . e antibacterial
activity of aqueous extract of W. somnifera against E. coli was
higher compared to that of the alcoholic extract . e
extracts of W. somnifera root were also eﬀective against
multidrug-resistant S. aureus , with methanol extract of
W. somnifera being also eﬀective in inhibiting oral bacteria,
like Streptococcus mutans and Streptococcus sobrinus .
Halamova et al.  investigated the antimicrobial activity of
W. somnifera against human pathogenic bacteria and ob-
served that those pathogens were more susceptible to extracts
compared to beneﬁcial Biﬁdobacteria. Interestingly, Zahran
et al.  also reported that the dietary supplementation with
W. somnifera root powder exhibited immunotherapeutic
activity against Aeromonas hydrophila in Nile tilapia.
When looking at the eﬀect of W. somnifera isolated
constituents, ﬂavonoids have shown excellent antimicrobial
eﬀects against C. albicans, S. aureus, Proteus mirabilis, E. coli,
and P. aeruginosa, although no eﬀects were noted against
Aspergillus ﬂavus or Aspergillus niger . Interestingly, the
minimum inhibitory concentration (MIC) of W. somnifera
methanol extract against C. albicans and Neisseria gonor-
rhoeaewas reported as 20 mg/mL and 0.5 mg/mL, while that
of water extract against N. gonorrhoeaewas 10 mg/mL .
In addition, W. somnifera glycoprotein revealed
antibacterial eﬀects against Clavibacter michiganensis subsp.
michiganensis and antifungal activity against A. ﬂavus,
Fusarium oxysporum, and Fusarium verticillioides . Also,
it was reported that W. somnifera can be utilized in the
synthesis of silver nanoparticles with excellent antioxidant,
antimicrobial, and anticancer potential [79–81].
6. Health-Promoting Effects
As previously mentioned, Withania has been used since a
long time ago for diﬀerent clinical purposes. In the tradi-
tional system of medicine, Withania somnifera has been
used for anti-inﬂammatory, anticancer, antioxidant, adap-
togenic, and antistress purposes, along with as an immu-
nomodulator. Moreover, it also exerts a positive inﬂuence on
endocrine, cardiorespiratory, and central nervous system
(CNS) levels. For instance, it was stated that W. somnifera is
a powerful help in cancer management, with good tolerance
. Recently, upon evaluating the clinical evidence base
and investigating the potential role of W. somnifera in
managing cognitive dysfunction, Ng et al.  found that
W. somnifera extract improved performance on cognitive
tasks, executive function, attention, and reaction time. It also
appears to be well tolerated, with good adherence and
minimal side eﬀects. Using standardized W. somnifera ex-
tracts or its bioactive ingredients, new and more eﬀective
medications to treat cognitive impairment could be pro-
duced . Notwithstanding, despite the broad spectrum of
preclinical data available, the number of clinical trials
performed using W. somnifera is markedly scarcer (Table 2).
7. Food-Pharma Industry: Safety and
W. somnifera has traditionally been available in the form of
capsules and powder, being most often sold as a supplement.
However, it can now be found in a variety of food products,
including ghee, honey, and kombucha. More recently,
W. somnifera has also been incorporated in baked goods,
juices, and beverages, respectively, sweets (candies/snacks),
and dairy products marketed as “Functional Foods” or
“Nutraceuticals.” e worth of note is that the amount of
W. somnifera in food can vary widely, where the addition of
powder can range from 1 to 10% depending on the product
(baked good vs. beverage). Also, levels of W.somnifera up to
5% have also been found to have acceptable sensorial fea-
Herbal cookies designed as functional foods have also
been developed with W. somnifera leaf powder, with the ﬁnal
product presenting with an acceptable color, taste, and
texture while maintaining an acceptable shelf-life .
Incorporating W. somnifera into foods can serve several
functions; for example, it can provide excellent antioxidant
and human health beneﬁts. Moreover, the addition of
W. somnifera to ghee (clariﬁed butter fat) was found to be an
eﬀective natural antioxidant to prevent oxidative degrada-
tion (less than synthetic antioxidant BHA, butylated
hydroxyanisole) apart from providing health-promoting
beneﬁts. e antioxidant activities evaluated were
Journal of Food Quality 7
Table 2: Health-promoting eﬀect of Withania somnifera.
Biological activity Dose/duration Study design/subjects Eﬀect References
500 mg dried aqueous
extract of roots and
leaves/twice a day for 14
randomized, crossover study (n�20
Decrease aortic pressure 
300 mg roots extract/day,
Prospective double-blind, randomized,
placebo-controlled trial (n�64 subjects
with a history of chronic stress)
Reduce cortisol levels and the
scores of stress-assessment scales 
500 mg powder capsule/
twice a day, twice a day, 30
Single-trial group (n�30 subjects with
Reduce fatigue symptoms,
improve workability and quality-
of-life dimension scores
120 mg root extract/day,
Double-blind placebo-controlled trial
(n�30 individuals with the obsessive-
Improve eﬀect in Yale-Brown
300 mg root extract/day
Clinical control-placebo trial (n�55
type II diabetics, under oral
Improvement in stress and
250 mg root ethanol
extract/twice a day, 6
study (n�39 subjects with generalized
anxiety disorder, mixed anxiety and
depression, panic disorder, and
adjustment disorder with anxiety)
Improvement in anxiety score
across time 
1000 mg standardized
root extract/day, 12 weeks
double-blind (n�66 patients with
depression and anxiety symptoms)
Improvement in depression
single-item and anxiety-
depression cluster scores and
500 mg standardized root
extract/day 8 weeks
(n�53 patients with bipolar disorder)
Improvement in auditory-verbal
working memory (digit span
250 mg dried aqueous
extract of roots and
leaves/twice daily, 14 days
Prospective, double-blind, placebo-
controlled, crossover (n�20 healthy
Improvement in the cognitive and
psychomotor performance 
300 mg root extract/twice
daily, 8 weeks
Prospective, randomized, double-
blind, placebo-controlled (n�50
healthy man and female adults)
Improvement in general memory
and executive function 
300 mg roots extract/
twice daily, 12 weeks
Randomized, double-blind, and
placebo-controlled (n�50 healthy
athletic male and/or female adult)
Enhances the cardiorespiratory
endurance, improvement in the
250 mg standardized root
extract/twice daily, 14
randomized, and placebo-controlled
(n�50 healthy men)
Increased velocity, power, and
maximum oxygen consumption 
500 mg standardized root
extract/day Sensoril®, 12
Randomized, double-blind, placebo-
controlled (n�40 healthy,
recreationally active men)
Improves upper- and lower-body
strength in active men 
1000 mg standardized
root extract/day, 10–14
(n�26 healthy men)
Increased mean pain threshold
250–125 mg standardized
root extract/twice daily,
Randomized, double-blind placebo-
controlled (n�16 patients with knee
joint pain and discomfort)
Reduced pain and disability scores
(both doses), and promoted a
better response (at a higher dose)
450 mg root extract/day,
crossover (n�42 patients with
Reduced severity pain and a
disability score 
2000 mg root extract/day
every 8 h during
nonrandomized comparative trial
(n�100 patients with breast cancer in
Reduce score Piper’s fatigue score
Reduced Schwartz’s cancer fatigue
score and improved quality-of-life
8Journal of Food Quality
β-carotene bleaching assay, DPPH assay, and Rancimat
method, and the doses evaluated were 1.0% and 0.5% (w/w)
for aqueous and ethanolic W. somnifera extract, respectively.
Perhaps not surprisingly, much food product development
research has focused on incorporating W. somnifera into
foods commonly consumed in India. Nonetheless, as foods
containing W.somnifera are becoming widely available,
increasing attention and consideration must be given to the
potential occurrence of adverse eﬀect(s) as a result of
7.1. From erapeutic to Safety Proﬁle. Animal and human
studies have been conducted to determine the potential
impact in the treatment of a wide range of diseases, including
but not limited to cancer, immunosuppressive diseases,
anxiety and depression, Parkinson’s disease (PD), and fer-
tility . Studies performed so far suggest that the con-
sumption of up to 100 mg per kg of body weight in a single
dosage or approximately 21g per day is safe. Typically, a
therapeutic dose is ≤10 g/day, so that a total intake can be
more closely controlled when consumed in a capsule form.
In an animal model, W. somnifera extract was given for 28
days at oral doses of 0, 500, 1000, and 2000 mg/kg body
weight, and data obtained suggest that the administration of
W. somnifera extract up to 2000 mg/kg/day did not trigger
adverse eﬀect .
Several review articles broadly cover various human
clinical trials suggesting that W. somnifera has no adverse
health eﬀects during long-term (≥one-year) administration
. For example, a group of 64 subjects aged from 18 to 54
received a 300 mg capsule of W. somnifera root extract for a
period of 60 days . Any incidences of adverse events were
comparable in the placebo-control group and W. somnifera
group, with the diﬀerence being not statistically signiﬁcant.
Another study investigated the use of W. somnifera in re-
productive issues; for that, a group of 41 men received a dose
of 4 tablets (500 mg each) 3 times/day (i.e., 6 g/day) con-
taining W. somnifera root powder through oral route after
intake of food for 60 days . e placebo (wheat powder)
received a tablet form, consisting of 4 tablets (500 mg each) 3
times/day (i.e., 6 g/day) (n�45). No adverse health eﬀects
were stated using the W. somnifera root powder.
e impact of W. somnifera root extract supplementa-
tion in muscle strength and recovery of 57 male subjects (18
to 50 years old) was also evaluated . Subjects in the
treatment group received 300 mg of W. somnifera root ex-
tract twice daily for 8 weeks, and no adverse health events
were reported. Taken together, data obtained so far appears
to support that W. somnifera has no toxic eﬀects; however,
such studies were not speciﬁcally designed to address safety
and adverse eﬀects. Also, most studies were of short duration
and, as such, may not be indicative of the long-term impact
of W. somnifera intake in human health.
7.2. Pregnancy and Teratogenicity. To what concerns, the
safe use of W. somnifera during pregnancy,whether as a
supplement or in food, remains uncertain. Reports suggest
that W. somnifera might have abortifacient properties
during pregnancy, indicating classiﬁcation under toxic
plants that cause abortion and sterility [122, 123]. In this
way, some researchers addressed the concern by orally
administering W. somnifera root extract to pregnant rats
during a period of major organogenesis and histogenesis
(days 5 to 19 of gestation). Brieﬂy, pregnant rats received a
dose of 500, 1000, and 2000 mg/kg/day and were monitored
for a range of clinical symptoms, although no evidence of
maternal or fetal toxicity was stated. e root extract pro-
voked no changes in body weight of parental females, the
number of corpora lutea, implantations, viable fetuses, and
external, skeletal, and visceral malformations. us, the
authors proposed evidence of safety related to W. somnifera
root extract at least at 2000 mg/kg/day . Regardless,
caution must be exercised concerning the use of
W. somnifera during pregnancy given the limited number of
published studies addressing the issue [122, 123]. According
to the National Institutes of Health , W. somnifera
contains several compounds that may cause miscarriage,
premature birth, or uterine contractions . W. somnifera
is commonly safely used by adults in doses up to 1000 mg per
day, for up to 12 weeks, but pregnant and breastfeeding
women should not consume .
Collectively, the wealth of research suggests that oral
intake of W. somnifera is safe with a possible exception
during pregnancy. In addition, given that W. somnifera is
being formulated into a wide range of commercially avail-
able food and beverages, the total day consumption by
consumers of such products may need to be more closely
considered. In this sense, future research may focus on
diﬀerences in bioavailability of the various forms (leaf and
root powder, extracts, and essential oils) related to safety and
e Withania genus has been traditionally used for its
therapeutic potential in numerous diseases, of which in-
somnia, depression, and immunostimulant eﬀects stand out.
However, remarkable anti-inﬂammatory and rejuvenating
activities have also been stated, with in vitro and in vivo
studies highlighting excellent antioxidant, antiproliferative,
cytotoxic, anti-inﬂammatory, and antimicrobial activity.
However, not all species present the same activity, with the
most studied and economically important one being the
roots of W. somnifera. More importantly, the clinical studies
performed so far have progressively aﬃrmed the
W. somnifera therapeutic eﬀects, namely, its excellent ability
to increase vitality, physical performance, and hematopoietic
capacity and to treat insomnia. Moreover, W. somnifera is
being valued for its ability to promote longevity and
strengthen the immune system without stimulating the
body’s reserves. Nonetheless, despite the advances stated so
far, further clinical trials and more precise and deeper
studies, namely, addressing the bioavailability and eﬀect of
pure compounds and the occurrence of synergistic eﬀects
when used in combination, along with the development of
methods to standardize the percentage composition of active
compound(s) in marketed products, are the ﬁelds that most
Journal of Food Quality 9
need to be intensively explored. Actually, although it is
possible to ﬁnd various products containing W. somnifera at
variable amounts and safety studies do not report adverse
eﬀects, it is of utmost importance to have deeper knowledge
on synergistic eﬀects that may possibly occur with other food
components and to know what are the eﬀects when high
doses are used and even what are the eﬀects in pregnancy.
e data supporting this review are from previously reported
studies and datasets, which have been cited. e processed
data are available from the corresponding author upon
Conflicts of Interest
e authors declare that they have no conﬂicts of interest.
All authors contributed equally to the manuscript. Con-
ceptualization was done by Javad Shariﬁ-Rad, Hari Prasad
Devkota, Beraat ¨
Ozçelik, Miquel Martorell, William C. Cho,
alia Cruz-Martins; Cristina Quispe, Seyed Abdul-
majid Ayatollahi, Farzad Kobarfard, Mariola Staniak, Anna
n, Katarzyna Czopek, Surjit Sen, Krishnendu Acharya,
Karl R. Matthews, Bilge Sener, Celale Kırkın, Montserrat
Victoriano, Deepak Chandran, Manoj Kumar, and Haﬁz
Ansar Rasul Suleria contributed to validation, investigation,
data curation, and writing the draft of the manuscript; re-
view and editing of the manuscript were performed by Javad
Shariﬁ-Rad, Hari Prasad Devkota, Beraat ¨
Martorell, William C. Cho, and Nat´
alia Cruz-Martins. All
authors read and approved the ﬁnal manuscript.
N. C. -M. acknowledges the Portuguese Foundation for
Science and Technology under the Horizon 2020 Program
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