Unravelling anti-cancer properties of solanaceous extracts using GC–MS and HPLC

Abstract

Cancer is a complex health issue that today’s medical science is dealing with, which has a mortality rate that is almost identical to that of cardiac disorders. Because of the adverse effects of the majority of the medications used in its therapy, managing it remains a major concern despite the availability of numerous remedies. This article attempts to contribute to the scientific developments in the Solanaceae family’s anti-cancer field. Thus, this study consisted of isolating β-amyrin, cedryl acetate, lupeol, and 2-pentadecanone, 6,10,14-trimethyl from Cestrum aurantiacum to determine the anti-tumor activity. The plant material was dried, pulverized, and small-scale extraction was done. Brine shrimps, cell lines (A549, Hela, HEPG), and Caenorhabditis elegans were used initially to examine three solanaceous plant extracts (Solanum villosum (SV), Cestrum aurantiacum (CA), and Brugmansia suaveolens (BS)). The best results were shown by ethanol extract of Cestrum aurantiacum that why large-scale extraction and GCMS of this extract were done. The antitumor potential can be explained by the presence of β-amyrin, cedryl acetate, lupeol, and 2-pentadecanone, 6,10,14-trimethyl.

Full text

Unravelling anti-cancer properties
of solanaceous extracts using GC–
MS and HPLC
Iqra Riaz1, Yamin Bibi1,2, Muhammad Arshad1, Muhammad Sheeraz Ahmad3,
Manzer H. Siddiqui4, Yawen Zeng5 & Abdul Qayyum6
Cancer is a complex health issue that today’s medical science is dealing with, which has a mortality
rate that is almost identical to that of cardiac disorders. Because of the adverse eects of the majority
of the medications used in its therapy, managing it remains a major concern despite the availability
of numerous remedies. This article attempts to contribute to the scientic developments in the
Solanaceae family’s anti-cancer eld. Thus, this study consisted of isolating β-amyrin, cedryl acetate,
lupeol, and 2-pentadecanone, 6,10,14-trimethyl from Cestrum aurantiacum to determine the anti-
tumor activity. The plant material was dried, pulverized, and small-scale extraction was done. Brine
shrimps, cell lines (A549, Hela, HEPG), and Caenorhabditis elegans were used initially to examine
three solanaceous plant extracts (Solanum villosum (SV), Cestrum aurantiacum (CA), and Brugmansia
suaveolens (BS)). The best results were shown by ethanol extract of Cestrum aurantiacum that why
large-scale extraction and GCMS of this extract were done. The antitumor potential can be explained
by the presence of β-amyrin, cedryl acetate, lupeol, and 2-pentadecanone, 6,10,14-trimethyl.
Keywords Cancer, Cestrum aurantiacum, GCMS, Lupeol
Abbreviations
CME Crude Methanol Extract
A549 Lung Cancer Cells
Hela Cervical Cancer Cells
HEPG Liver cancer cells
SV Solanum villosum
CA Cestrum aurantiacum
BS Brugmansia suaveolens
GCMS Gas Chromatography Mass Spectrometry
ICMR Indian Council of Medical Research
TCM Traditional Chinese Medicine
HPLC High-performance liquid chromatography
DMSO Dimethyl sulfoxide
PMAS Pir Mehr Ali Shah
BSLT Brine shrimp lethality test
MTT 3-(4, 5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
NGM Nematode Growth Medium
C. elegans Caenorhabditis elegans
E. coli OP50 culture Escherichia coli OP50 culture
HPLC-DAD High-performance liquid chromatography with diode-array detection
ROS Reactive oxygen species
1Department of Botany, PMAS-Arid Agriculture University Rawalpindi, Rawalpindi 46300, Pakistan. 2Department
of Botany, Rawalpindi Women University, Rawalpindi 46300, Pakistan. 3University Institute of Biochemistry and
Biotechnology, PMAS-Arid Agriculture University Rawalpindi, Rawalpindi 46300, Pakistan. 4Department of Botany
and Microbiology, College of Science, King Saud University, 11451 Riyadh, Saudi Arabia. 5Biotechnology and
Germplasm Resources Institute, Agricultural Biotechnology Key Laboratory of Yunnan Province/Key Laboratory
of the Southwestern Crop Gene Resources and Germplasm Innovation, Ministry of Agriculture, Yunnan Academy
of Agricultural Sciences, Kunming 650205, Yunnan, China. 6Department of Agronomy, The University of Haripur,
Haripur 22620, Pakistan. email: yamin.bibi@f.rwu.edu.pk; aqayyum@uoh.edu.pk
OPEN
Scientic Reports | (2025) 15:4192 1
| https://doi.org/10.1038/s41598-025-87654-9
www.nature.com/scientificreports
Content courtesy of Springer Nature, terms of use apply. Rights reserved

Cancer is the second leading cause of death worldwide aer heart disorders. Every year, about 6 million individuals
lose their lives to cancer, out of 10 million diagnoses1. Lung, mouth, stomach, and esophageal cancers are more
common in men, but breast, uterine, and cervical cancers are more common in women, according to e National
Cancer Registry Programme Report, 2020 of Indian Council of Medical Research (ICMR). According to estimates,
the number of cancer cases will probably increase from 13.9 lakh in 2020 to 15.7 lakh by 2025. Even though cancer
patients now have more options because of advancements and innovation in the eld, it is crucial to provide anti-
cancer drugs that are non-toxic, widely accessible, and reasonably priced2. e anti-cancer potential of over 3000
plants has been revealed worldwide3. Currently, natural plant products account for around 60% of the anti-cancer
drugs now in use4. It’s interesting to note that 90 of the approximately 120 medications used to treat cancer come
from plants2.
Solanaceae is one of the diverse plant families of the 405 angiosperm groups, with a wide range of benecial
chemical components5. e Solanaceae family is signicant for both medicinal and economic plants6.
Conversely, some of the most signicant Solanaceae plants inuenced the early phases of medical plant-based
drug development and are still valued in herbal medicine7. ere are over 2700 species in the Solanaceae family,
divided into 98 genera8. Numerous traditional medical systems, such as Ayurveda, Traditional Chinese Medicine
(TCM), Siddha, Unani, and homeopathy, employ some of the alkaloids found in the Solanaceae family7,9.
Considering the enormous anti-cancer potential of Solanaceae plants, we designed the current investigation
by selecting three Solanaceae plants (Solanum villosum (SV), Cestrum aurantiacum (CA), and Brugmansia
suaveolens (BS) to search for anti-cancer compounds.
Leaves of Brugmansia suaveolens are traditionally applied in northern Peru for treating wounds10. Flowers and
leaves are used for curing infections, mental weaknesses, and menstrual pain. It is utilized as an aphrodisiac11.
e stem is applied externally for skin anomalies for rapid healing12. Brugmansia has hallucinogenic eects,
it also has anti-inammatory, antirheumatic, and analgesic properties13. Solanum villosum leaves and fruits
are eaten or made into decoctions to cure stomach problems like dysentery and diarrhea. is is common in
traditional African medicine14. e plant is frequently eaten as a vegetable, particularly in East Africa, where its
high nutrient prole is thought to support immunity and overall health15. Applying crushed leaves straight to
cuts and wounds speeds up the healing process. e antibacterial qualities of the plant are thought to aid in the
prevention of infections. It also has anti-inammatory properties16. Traditional use of extracts supports their use
in treating a variety of illnesses due to their antibacterial and antifungal properties17.
Genus Cestrum comprises more than 300 species and is widely distributed in tropical and subtropical
areas around the world like Bangladesh, India, the United States, Australia, South America and southern
China.18,19 Cestrum species have a long history in folk medicine for the treatment of several diseases and health
disorders.18,20Dierent species belonging to the Cestrum genus were previously investigated for their chemical
constituents, e.g.,parquine, carboxyparquine and steroids. Glycosides were also identied in Cestrum diurnum.21
whereas saponins were identied in Cestrum parqui.22,23.
A variety of bioactive chemicals found in Cestrum plants have been linked to their cytotoxic and antitumor
eects. e pharmacological actions of Cestrum species are attributed to the presence of avonoids, alkaloids, and
saponins, as revealed by phytochemical research.19,24. Due to the substantial cytotoxic eects these compounds
have demonstrated against various cancer cell lines in vitro, Cestrum species are promising candidates for
developing anticancer drugs. For instance, Nasr et al. 24 studied extracts from two Cestrum species and found
promising cytotoxic eects against certain cancer cells, conrming the traditional use of these plants for medical
purposes.
Cestrum aurantiacum (Orange cestrum) is a plant used for various medicinal purposes in traditional South
Asian medicine, especially Ayurveda. e plant is mostly known for its therapeutic properties in many traditional
healing methods. Roots and leaves have been used for their anti-inammatory properties. Occasionally, they are
used as decoctions or applied as poultices to relieve joint pain, muscle pains, and swelling. It has antimicrobial
properties and is frequently used to treat fungal diseases, wounds, and skin infections. It can also be used to
relieve constipation, bloating, and stomach pain. It may be used as a traditional remedy for respiratory conditions
including colds and coughs. Occasionally, preparations that aid in clearing respiratory passages contain it. e
plant has been used in some cases as a febrifuge to help reduce fever, although it has to be used with caution
as certain parts of the plant can be poisonous when consumed in the wrong way. e plant is oen used in
controlled doses in traditional practices and caution is advisable when using it as a medicine.25.
e current study aimed to use bioassay-guided purication to extract the main antitumor components of
solanaceous plant species.
Materials and methods
Reagents
Solvents of the high-performance liquid chromatography (HPLC) grade, such as acetone, methanol, acetonitrile,
hexane, ethanol, Deionized water and DMSO were used.
Collection and identication of plants
From spring 2017 to spring 2018, solanaceous plant species were collected from dierent areas of Pakistan.
Solanum villosum was collected from Narowal, province of Punjab. Cestrum aurantiacum and Brugmansia
suaveolens were collected from Islamabad. Prof. Dr. Rahmatullah Qureshi, identied the specimens and
voucher specimens were stored for future use as reference (Voucher # 168, 221, 339). Aer the plants were
completely dried, a ne powder was made and preserved. All research studies and methods were carried out
per institutional, national, and international guidelines and legislations. We got the appropriate permissions and
licenses for the collection of plant specimens from the Administration Department, PMAS-Arid Agriculture
University Rawalpindi, and the Department of Agriculture, Government of Punjab, Pakistan.
Scientic Reports | (2025) 15:4192 2
| https://doi.org/10.1038/s41598-025-87654-9
www.nature.com/scientificreports/
Content courtesy of Springer Nature, terms of use apply. Rights reserved

Small-scale extraction
e plant material that had completely dried out was ground into a powder, and by utilizing a technique designed
by Panda et al.26, small-scale extraction was accomplished. Ten milliliters of solvent (hexane, chloroform,
Acetone, ethanol, and water) was used with 1g of plant powder (S. villosum, C. aurantiacum, and B. suaveolens).
Aer every four hours, the tubes were put in a sonicator bath for one hour. One milliliter of aliquots from
each fraction was dried in a Savant SpeedVac Concentrator 200H.
Cytotoxic activities
Brine shrimp lethality assay
For brine shrimp assay cysts of Artemia salina were hatched in two-chambered containers with small holes in
the partitioning wall. For seawater 38g/L sea salt was taken in a beaker and oxidation of this water was done
by using a magnetic stirrer for approximately 2h. Aer approximately 48h’ cysts hatched and swam through
the hole to the illuminated chamber, from where shrimps were collected for an experiment. e tested extracts
were dissolved in 100% DMSO and used as a stock solution at a concentration of 60mg/mL. Utilizing articial
seawater, dierent concentrations were prepared from stock. Articial seawater was employed as a negative
control, and nicotine was used as a positive control. Ten phototropic nauplii were introduced to each container
and cultured for 24h at room temperature. Dead nauplii were counted and the percentage of lethality was
calculated aer 24h.27.
Percentage of Death = (Total nauplii−Alive nauplii) / Total nauplii × 100.
In-vitro cytotoxicity assay (MTT assay)
e KU Leuven Zoology Department’s Animal Physiology and Neurobiology lab provided the cell lines. e
cells were stored in liquid nitrogen stock, awing was done before use. Aer thawing, the cells were moved to
a sterile falcon tube, and 5mL of cell culture medium was added. e density of the cells was examined under
a microscope, aer which 2mL of cell culture had been transferred. A cell culture ask was lled with 15mL of
Dulbecco’s Modied Eagle’s Medium (DMEM). e ask was then incubated at 37°C for 24h in an incubator
with 5 percent CO2 to allow the cell to attach to and develop on the ask’s surface.
In a ask, 5mL of complete media was added to trypsinized cells. In a 15mL falcon tube, centrifuged for
5min at 500rpm. e medium was withdrawn, and cells were re-suspended in complete media (1.0mL). Cells
were counted, and full media was used to dilute it to 75,000 cells per mL in a 96-well plate. Fill each well with 100
µL of cells (7500 total cells) and set it in a CO2 incubator overnight. e next day, tested extracts were added, and
the ultimate amount was held at 100 µL each well. Each well received 20 µL of 5mg/mL MTT. As a control, MTT
was added to one set of wells with no cells. ese plates were incubated in a CO2 incubator for 3.5h at 37°C.
Absorbance was measured at 590nm using a 620nm reference lter.28.
C. elegans lethality assay
C. elegans were cultured on Nematode Growth Medium (NGM). e N2 wild-type C. elegans strain was utilized.
C. elegans were treated to synchronization at the L4 larval stage. 96-well microplate with a at bottom from TPP
Techno Plastic Products AG in Switzerland was used. A 96-well microplate with 189 μL of E. coli OP50 culture
(OD = 0.5 at 620nm) was lled with synchronized C. elegans 10 μL, or roughly 40 to 45 numbers of L4 larvae.
Plant extract in the amount of 1 μL was tested; 1 μL of DMSO was used as a solvent control, and 1 μL of 50μM
Levamisole was used as a positive control. Worms and extracts were combined, and then a 96-well microplate
was put into a WMicroTracker (Phylumtech, Argentina) device and incubated for 24h at 20°C. Every 30min,
the WMicroTracker measured and recorded the worm movement in each well. To determine the relative activity,
the percentage of the average movement over 24h of test samples with extract compared to the DMSO control
was employed.29.
Large-scale extraction
For large-scale extraction, a big container containing 2000mL of ethanol HPLC grade from Sigma (Germany)
was lled with 200g of plant powder (C. aurantiacum). e container was kept at room temperature for a
full day. To increase the extraction yield, the container was submerged in a water bath sonicator four times
for a duration of sixty minutes each. ere was a minimum 6-h pause between each sonication to allow the
suspension to reach room temperature.
e plant material was ltered using VWR Grade 313 size 5lm lter paper aer the fourth sonication to
obtain the dry residue. e ltrate was then evaporated using a rotary evaporator (BUCHI rotavapor R-100), and
the weight of the rst dried extract was determined. e recovered ethanol was utilized once more for a second
extraction using the same protocol for twenty-four hours. e ltrate was then again evaporated using a rotary
evaporator to obtain the dry extract.
To extract every ingredient from the plant powder, the same process was carried out repeatedly. e dried
material’s nal weight was computed. For additional examination, the plant extract was kept in storage at 4°C.
Purication
A silica gel was used to bind the plant extract, and a silica column was made. Hexane, ethyl acetate, methanol,
and acetic acid were blended into eluates using a step gradient that increased in polarity. Finally, 100% acetic
acid was used to elute the column.
Reverse-phase high-performance liquid chromatography analysis
A Shimadzu LC-20AT system with an LC-20AT quaternary pump and an online degasser (DGU20A3/DGU-
20A5) was used to perform high-performance liquid chromatography (HPLC–DAD) analyses. Lab Solution
Scientic Reports | (2025) 15:4192 3
| https://doi.org/10.1038/s41598-025-87654-9
www.nature.com/scientificreports/
Content courtesy of Springer Nature, terms of use apply. Rights reserved

soware was used to obtain the data. e HPLC column was in the reverse phase. e mobile phase consisted of
acetonitrile, water, and 0.1% triuoroacetic acid (TFA).
Gas chromatography-mass spectrometry
e peaks that were collected were put via gas chromatography. Using an internal diameter of 0.18mm and
a thickness of 0.18mm, a Restek RXi-5sil MS 20m column was employed, and the NIST 14 MS library was
utilized to search the spectrum.
Results
Brine shrimps, cell lines (A549, Hela, HEPG), and C. elegans were used initially to examine three solanaceous
plant extracts (S. villosum (SV), C. aurantiacum (CA), and B. suaveolens (BS)). e extract of C. aurantiacum was
shown to have a broader spectrum of ecacy against all the tested activities. e cytotoxic activity of all three
solanaceous plant extracts was examined by using dierent solvents from low polarity to high polarity (hexane,
chloroform, acetone, ethanol, and water). e ndings indicated that the most active ingredients were in the
ethanol extract of C. aurantiacum (Figs.1, 2 and 3).
erefore, ethanol was used to conduct a large-scale extraction, and the resulting extract was then further
divided into 220 fractions using column chromatography (Fig.4). All fractions were evaluated for activity
against Hela cell line; fractions 31–38, 54–57 and 122–128 were conrmed to be active (Fig.5). Aer being
dried, the active portions were weighed. A serial dilution approach was used to test the active fractions against
the HeLa cell line once more (Fig.6). e results showed that fractions 31–38 consistently exhibited cytotoxic
activity, suggesting the presence of a potent active antitumor component. Consequently, fractions 34 and 37
underwent further separation with the use of a mobile phase comprising acetonitrile and water. Hela cells were
used to evaluate all 60 subfractions once more, and the results showed that there were two active subfractions in
each fraction (Fig.7 and 8). For identication, active peaks were subjected to GCMS (Fig.9, 10, 11 and 12). It
was determined that compound 1 was β-amyrin, compound 2 was cedryl acetate, compound 3 was lupeol, and
compound 4 was 2-pentadecanone, 6,10,14-trimethyl.
Discussion
ere are around 300 species of the genus Cestrum (Family Solanaceae), which is extensively spread in
tropical and subtropical regions of the world, including Bangladesh, India, the United States, Australia, South
America, and southern China.18,19 In traditional medicine, Cestrum species have long been used to treat
a variety of illnesses and ailments.18,20 In Chinese traditional medicine the leaves of C. nocturnum to cure a
variety of ailments. e plant exhibited biological actions that were detectable, such as antitumor, antioxidant,
hepatoprotective, cytotoxic, antibacterial, anticonvulsant, larvicidal, anti-inammatory, analgesic, antitumor,
and wound healing. Previous studies on the chemical composition of various Cestrum species, such as parquine,
carboxyparquine, and steroids, were conducted. While C. parqui was found to contain saponins, C. diurnum was
Fig. 1. Mean % lethality of brine shrimp in dierent solvents of selected plant species.
Scientic Reports | (2025) 15:4192 4
| https://doi.org/10.1038/s41598-025-87654-9
www.nature.com/scientificreports/
Content courtesy of Springer Nature, terms of use apply. Rights reserved

also shown to contain glycosides.24 Cestrum aurantiacum was least investigated, β-amyrin, cedryl acetate, lupeol,
and 2-pentadecanone, 6,10,14-trimethyl were isolated from this species.
β-amyrin has been identied from dierent plants i-e from 5 varieties of Humulus lupulus30. Celastrus
hindsii leaves31, Alstonia boonei32 and several materials have been found to contain amyrin, including an
ethanolic fraction of the oleogum resin from Ferula gummosa33, methanolic extract of Carpobrutus edulis34
Chloroform extract of Euphorbia tirucalli L.35leaves of Ficus benjamina36 leaves of Pyrenacantha staudii,37 Olea
Fig. 2. Inhibitory eects (%) of selected plants on various cell lines using dierent solvents.
Scientic Reports | (2025) 15:4192 5
| https://doi.org/10.1038/s41598-025-87654-9
www.nature.com/scientificreports/
Content courtesy of Springer Nature, terms of use apply. Rights reserved

europea L. leaves (ethanol extract)38. Leaves of Tectona philippinensis and Rhus alata39 and Piptadenia Africana
stem bark also contain β-Amyrin40.
e chemical structure of β-amyrin (3β- hydroxy-olean-12-en-3-ol) is also depicted in Figure and its formula
is C30H50O. e infra-red spectrum of β-amyrin shows the presence of a hydroxyl function and the olenic
moiety at a spectrum of 3360 and 1650cm-1 and MS studies of β-amyrin conrm a parent ion peak at m/z
426 (M +)41, other work of HR-EI-MS m/z: 426.2975 (calcd. for C30H50O, 426.3861) (Jabeen et al., 2011). e
melting point of β-amyrin is 189–191ºC42.
Cedryl acetate is a sesquiterpene compound, the pure chemical is crystalline, which is derived from
natural oils found to be volatile in nature. A tricyclic sesquiterpene called cedryl acetate was discovered in
Fig. 4. An overlay chromatogram of an ethanol extract of Cestrum aurantiacum from a silica gel column;
fractions were taken per minute.
Fig. 3. Percentage lethality of C. elegans in dierent solvents of selected plant species.
Scientic Reports | (2025) 15:4192 6
| https://doi.org/10.1038/s41598-025-87654-9
www.nature.com/scientificreports/
Content courtesy of Springer Nature, terms of use apply. Rights reserved

the plant Psidium caudatum. Cedryl acetate exhibits antiglucosidase action. Cedryl acetate is also present in
Cunninghamia lanceolata var. konishii.43 In vitro anticancer, and antimicrobial activities of the Heartwood
Essential Oil of Cunninghamia lanceolata var. konishii was evaluated from Taiwan, the oil exhibited cytotoxic
activity against human lung, liver and oral cancer cells.44 Crataegus AzarolusL has Cedryl acetate so evaluated
for its antioxidant, Anti-Inammatory, and Antiproliferative Activities.45 It has Density 0.999g/mL at 25°C.
Melting point 44–46℃. e boiling point is 291.7°C at 760mmHg.43.
A hydroxy group replaces the hydrogen at the 3beta position of the pentacyclic triterpenoid lupane to form
lupeol, which is present in fruits and vegetables, preferentially caused signicant head and neck squamous cell
carcinoma (HNSCC) cell death in vitro but had little to no impact on a normal tongue broblast cell line.
e primary mechanism of lupeol’s anticancer eects against HNSCC has been found to be down-regulation
of NF-kappaB. Lupeol alone was discovered to reverse the NF-kappaB-dependent epithelial-to-mesenchymal
transition, which not only reversed tumor growth but also impaired HNSCC cell invasion. Lupeol and cisplatin
together had a synergistic impact that in vitro chemosensitized HNSCC cell lines with signicant NF-kappaB
activity46. Previously it has been reported from dierent plants like Derris scandens, Albizia procera, and
Diospyros rhodocalyx.47 Nyctanthes arbor-tristis was also invested for lupeol compound.48.
2-pentadecanone, 6,10,14-trimethyl is present in the aerial parts of Andrographis paniculata49 leaves of
Combretum latifolium50 Crinum latifolium L.51 and Curcuma aromatica Salisb52 while in the ower of Jasminum
graniorum53 and also in the Rumex vesicarius54. is substance is a member of the sesquiterpenoids class of
organic substances. ese terpenes have three isoprene units following one another with density 0.590336 and
Electronegativity -4.861.
Correlation between identied compounds and the investigated biological activities
β-Amyrin and biological activities
According to Sandeep et al.55, β-amyrin demonstrated cytotoxicity in brine shrimp lethality assays, a standard
screening method for bioactive compounds. is cytotoxicity may be attributed to its ability to disrupt cellular
membranes and induce apoptosis in these organisms. A study by Moy et al.56showed that β-amyrin induces
apoptosis in A549 cells, reducing cell proliferation and survival. e apoptosis-inducing ability of β-amyrin in
A549 cells supports its potential as an anticancer agent. Gautam et al.57highlighted the anticancer properties of
β-amyrin, demonstrating its ability to inhibit cell growth and induce cell cycle arrest in HeLa cells. is action
in HeLa cells is linked to β-amyrin’s anti-inammatory and apoptosis-inducing properties.Bhuvaneshwaran
et al.58reported that β-amyrin suppresses liver cancer cell proliferation and induces apoptosis through ROS
generation. Its ability to induce oxidative stress makes it eective in liver cancer cells, corroborating its anticancer
action. e antioxidant activity of β-amyrin was demonstrated in C. elegans by Rani et al.59, where it was shown
to protect against oxidative stress. is suggests that β-amyrin may oer protective eects beyond cancer studies
to stress and aging-related models in C. elegans.
Cedryl acetate and biological activities
Cedryl acetate is a sesquiterpene ester with antimicrobial, anti-inammatory, and potential anticancer activities.
Subramanian et al.60found that cedryl acetate exhibited cytotoxic eects in brine shrimp assays, suggesting
it may disrupt cellular integrity Cedryl acetate’s cytotoxic eects in brine shrimp may be related to its ability to
Fig. 5. e cytotoxic activity of the Cestrum aurantiacum fractions obtained by the silica column.
Scientic Reports | (2025) 15:4192 7
| https://doi.org/10.1038/s41598-025-87654-9
www.nature.com/scientificreports/
Content courtesy of Springer Nature, terms of use apply. Rights reserved

induce apoptosis or cell membrane disruption. In a study by Sharma et al.61, cedryl acetate was shown to have
anti-inammatory eects in lung cancer models. Its anti-inammatory properties could contribute to reducing
lung cancer cell proliferation, possibly by inhibiting inammatory pathways that promote cancer growth. Prasad
et al.62reported that cedryl acetate has anti-cancer eects by inducing apoptosis in HeLa cells Cedryl acetate’s
potential to induce apoptosis in cervical cancer cells further supports its use as a therapeutic agent. Reddy et
al.63showed that cedryl acetate has antioxidant properties that could inuence liver cancer cells by mitigating
oxidative damage is suggests cedryl acetate may modulate oxidative stress pathways involved in liver cancer
Fig. 6. Cytotoxic activity of most active fractions of Cestrumaurantiacum against hela cell line using a twofold
dilution protocol.
Scientic Reports | (2025) 15:4192 8
| https://doi.org/10.1038/s41598-025-87654-9
www.nature.com/scientificreports/
Content courtesy of Springer Nature, terms of use apply. Rights reserved

progression.e antimicrobial activity of cedryl acetate was studied by Das et al.64, where it showed potential
toxicity to C. elegans under laboratory conditions: Its toxicity could be related to the disruption of cellular
functions in C. elegans, possibly through membrane destabilization.
Lupeol and biological activities
Lupeol is a triterpene known for its anticancer, anti-inammatory, and antioxidant eects. Shah et al.65found
that lupeol exhibited signicant cytotoxicity in brine shrimp assays Lupeol’s cytotoxicity is likely due to its ability
to induce apoptosis or interfere with cellular metabolism. Tripathi et al.66 demonstrated that lupeol inhibits
A549 cell proliferation and induces apoptosis via the activation of caspase-3 and -9. Lupeol’s apoptosis-inducing
activity in A549 cells is key to its anticancer eects.Borse et al.67 showed that lupeol has potent anti-proliferative
and apoptotic eects in HeLa cells, reducing cell viability and migration. Lupeol’s anticancer action is further
conrmed in cervical cancer models by inhibiting growth and metastasis. Anwar et al.68 found that lupeol
suppresses liver cancer cell proliferation by inhibiting the PI3K/AKT signaling pathway. Lupeol’s inhibition of
critical signaling pathways in liver cancer cells supports its potential as an anti-liver cancer agent.Ali et al.69
reported that lupeol extends lifespan and promotes antioxidant activity in C. elegans, indicating its protective
eects against oxidative stress. e ability of lupeol to protect C. elegans from oxidative damage further supports
its antioxidant and health-promoting eects.
2-Pentadecanone, 6,10,14-trimethyl and biological activities
2-Pentadecanone, 6,10,14-trimethyl is a ketone compound known for its antimicrobial and possible anticancer
eects. Chavan et al.70 reported that 2-pentadecanone exhibited cytotoxicity in brine shrimp lethality tests. Its
cytotoxicity is linked to its ability to disrupt cellular membranes or induce apoptosis.Sharma et al.71 found that
Fig. 7. (a) HPLC chromatogram of fraction 34 (Cestrum aurantiacum); fractions were collected per minute.
(b) Tested for activity (percentage inhibition of cancer cells).
Scientic Reports | (2025) 15:4192 9
| https://doi.org/10.1038/s41598-025-87654-9
www.nature.com/scientificreports/
Content courtesy of Springer Nature, terms of use apply. Rights reserved

Fig. 9. Mass spectra of Compound 1 (β-Amyrin) and its structure.
Fig. 8. (a) HPLC chromatogram of fraction 37 (Cestrum aurantiacum); fractions were collected per minute.
(b) Fractions tested for activity (percentage inhibition of cancer cells).
Scientic Reports | (2025) 15:4192 10
| https://doi.org/10.1038/s41598-025-87654-9
www.nature.com/scientificreports/
Content courtesy of Springer Nature, terms of use apply. Rights reserved

2-pentadecanone inhibits the growth of A549 cells by inducing cell cycle arrest and apoptosis. e compound’s
ability to halt cell cycle progression and induce apoptosis makes it a potential anticancer agent.
Kumar et al.72 suggested that 2-pentadecanone shows promising anti-cancer activity in HeLa cells by altering
cell morphology and inducing cytotoxicity. e anti-cancer eect in HeLa cells indicates that this compound may
interfere with cellular integrity or metabolism.Venkatesan et al.73 showed that 2-pentadecanone has potential
anticancer properties in liver cancer models, possibly by inducing oxidative stress and apoptosis. Its oxidative
stress-inducing properties align with its role in liver cancer treatment. In C. elegans, 2-pentadecanone showed
toxicity, possibly by aecting metabolic pathways or inducing cellular stress. Its toxicity in C. elegans highlights
its potential for further study in toxicity and metabolic disruption.
Fig. 12. Mass spectra of Compound 4 (2-Pentadecanone, 6,10,14-Trimethyl) and its structure.
Fig. 11. Mass spectra of Compound 3 (Lupeol) and its structure.
Fig. 10. Mass spectra of Compound 2 (Cedryl Acetate) and its structure.
Scientic Reports | (2025) 15:4192 11
| https://doi.org/10.1038/s41598-025-87654-9
www.nature.com/scientificreports/
Content courtesy of Springer Nature, terms of use apply. Rights reserved

Across several model systems, these studies demonstrate a high correlation between the biological activities
of the substances β-amyrin, cedryl acetate, lupeol, 2-pentadecanone, and 6,10,14-trimethyl. eir potential as
anticancer medicines are suggested by their shared capacity to induce apoptosis, inhibit cell proliferation, and
function as antioxidants. ese correlations, which were derived from various research papers, highlight the
therapeutic value of these compounds in cancer treatment.
Conclusion
is study highlights the anticancer potential of ethanol extracts fromCestrum aurantiacum, with the identication
of key bioactive compounds such as 2-pentadecanone, 6,10,14-trimethyl; lupeol; cedryl acetate; and β-amyrin.
Notably, lupeol and 2-pentadecanone, 6,10,14-trimethyl are reported for the rst time in the genusCestrum,
while β-Amyrin marks its debut fromCestrum aurantiacum. ese ndings underscore the untapped therapeutic
potential of this plant and its unique phytochemical prole. Moving forward, the exploration of these compounds
through advanced biological assays, molecular docking, and in vivo studies could provide deeper insights into their
mechanisms of action against cancer. Furthermore, integrating modern approaches like nanocarrier systems for
targeted delivery and AI-driven compound screening could propel these bioactives toward clinical relevance. is
work lays a foundation for the future of plant-based oncological therapeutics, bridging traditional knowledge with
cutting-edge science.
Data availability
Data available on request from the corresponding author (Yamin Bibi).
Received: 19 September 2024; Accepted: 21 January 2025
References
1. Hanif, M., Zaidi, P., Kamal, S. & Hameed, A. Institution-based cancer incidence in a local population in Pakistan: Nine-year data
analysis. Asian Pac. J. Cancer Prev. 10, 227–230 (2009).
2. Rohilla, P., Jain, H., Chhikara, A., Singh, L. & Dahiya, P. Anticancer potential of Solanaceae plants: A review. South Afr. J. Bot. 149,
269–289 (2022).
3. Tariq, A. et al. A systematic review on ethnomedicines of anti-cancer plants. Phytother. Res. 31, 202–264 (2017).
4. Gordaliza, M. Natural products as leads to anticancer drugs. Clin. Transl. Oncol. 9, 767–776 (2007).
5. Fatur, K. “Hexing herbs” in ethnobotanical perspective: A historical review of the uses of anticholinergic Solanaceae plants in
Europe. Econ. Bot. 74, 140–158 (2020).
6. Ghatak, A. et al. Proteomics survey of the Solanaceae family: Current status and challenges ahead. J. Proteomics 169, 41–57 (2017).
7. Chowanski, S. et al. A review of bioinsecticidal activity of Solanaceae alkaloids. Toxins 8, 1–28 (2016).
8. Olmstead, R. G. & Bohs, L. A summary of molecular systematic research in Solanaceae: 1982–2006. In VI International Solanaceae
Conference: Genomics Meets Biodiversity 745 (International Society for Horticultural Science, Belgium, 255–268 (2006).
9. Shah, V. V., Shah, N. D. & Patrekar, P. V. Medicinal plants from Solanaceae family. Res. Pharm. Technol. 6, 143–151 (2013).
10. De Feo, V. Ethnomedical eld study in northern Peruvian Andes with particular reference to divination practices. J. Ethnopharmacol.
85, 243–256 (2003).
11. Descola, P. Phytochemical and antibacterial studies on Jatropha curcas L. J. Chem. Pharm. Res. 4, 2639–2642 (1996).
12. Ratsch, C. Enziklopädie der Psychoaktiven Panzen (AT Verlag, 1998).
13. Nencini, C. et al. Anity of Iresine herbstii and Brugmansia arborea extracts on dierent cerebral receptors. J. Ethnopharmacol.
105, 352–357 (2006).
14. Chandra, S. et al. Ethnomedicinal plants of the Solanum genus: A review. J. Med. Plants Res. 8, 457–468 (2014).
15. Grubben, G. J. H. & Denton, O. A. Plant Resources of Tropical Africa: Vegetables. PROTA Foundation (2004).
16. Kokwaro, J. O. Medicinal Plants of East Africa (University of Nairobi Press, 2009).
17. Oudhia, P. Traditional medicinal uses of Solanum species. Ethnobot. Leaets 5, 1–4 (2001).
18. Begum, A. S. & Goyal, M. Research and medicinal potential of genus Cestrum (Solanaceae)—A review. Pharmacogn. Rev. 1, 320
(2007).
19. Prasad, M. P., Apoorva, P., akur, M. S. & Ruparel, Y. M. Phytochemical screening, antioxidant potential, and antimicrobial
activities in three species of Cestrum plants. Int. J. Pharm. Biol. Sci. 14, 673–678 (2013).
20. Al Raza, S. M., Rahman, A. & Kang, S. C. Chemical composition and inhibitory eect of essential oil and organic extracts of
Cestrum nocturnum L. on food borne pathogens. Int. J. Food Sci. Technol. 44, 1176–1182 (2009).
21. Durand, R., Figueredo, J. M. & Mendoza, E. Intoxication in cattle from Cestrum diurnum. Vet. Hum. Toxicol. 41, 267–268 (1999).
22. Chaieb, I., Kamel, B., Trabelsi, M., Hlawa, W., Raouani, N., Ben Ahmed, D. et al. Pesticidal potentialities of Cestrum parqui
saponins. Int. J. Agric. Res. 2, 275–281 (2007).
23. Najet, R., Dorsaf, B. A., Mejda, D. & Habib, B. H. Pesticidal potentialities of Cestrum parqui saponins. Int. J. Agric. Res. 2, 275–281
(2007).
24. Nasr, S. M. et al. High performance liquid chromatography ngerprint analyses, in vitro cytotoxicity, antimicrobial and antioxidant
activities of the extracts of two Cestrum species growing in Egypt. Pharmacogn. Res. 10, 173 (2018).
25. Sivarajan, V. V. & Balachandran, I. Ayurvedic Drugs and eir Plant Sources (IBH Publishing Co, Oxford, 1994).
26. Panda, S. K. et al. Antimicrobial, anthelmintic, and antiviral activity of plants traditionally used for treating infectious diseases in
the similipal biosphere reserve, Odisha India. Front. Pharmacol. 8, 658 (2017).
27. Sasidharan, R. & Gerstein, M. Protein fossils live on as RNA. Nature 453, 729–731 (2008).
28. Joo, T. et al. Inhibition of nitric oxide production in LPS-stimulated RAW 264.7 cells by stem bark of Ulmus pumila L.. Saudi J. Biol.
Sci. 21, 427–435 (2014).
29. Panda, S. K., Das, R., Mai, A. H., De Borggraeve, W. M. & Luyten, W. Nematicidal activity of Holigarna caustica (Dennst.) Oken
fruit is due to linoleic acid. Biomolecules 10, 1043–1049 (2020).
30. Rokicka, K. & Wojciak-Kosior, M. Identication and quantitative analysis of 161 amyrins in Humulus lupulus L. Curr. Issues
Pharm. Med. Sci. 27, 142–144 (2015).
31. Viet, T. D., Xuan, T. D. & Anh, L. H. α-Amyrin and β-Amyrin isolated from Celastrus hindsii leaves and their antioxidant, anti-
xanthine oxidase, and anti-tyrosinase potentials. Molecules 26, 7248–7256 (2021).
32. Okoye, N. N. et al. β-Amyrin and α-amyrin acetate isolated from the stem bark of Alstonia boonei display profound anti-
inammatory activity. Pharm. Biol. 52, 1–10 (2014).
Scientic Reports | (2025) 15:4192 12
| https://doi.org/10.1038/s41598-025-87654-9
www.nature.com/scientificreports/
Content courtesy of Springer Nature, terms of use apply. Rights reserved

33. Jalali, H. T., Ebrahimian, Z. J., Evtuguin, D. V. & Neto, C. P. Chemical composition of oleo-gum-resin from Ferula gummosa. Ind.
Crops Prod. 33, 549–553 (2011).
34. Martins, A. et al. Constituents of Carpobrotus e dulis inhibit P-glycoprotein of MDR1-transfected mouse lymphoma cells. Anticancer
Res. 30, 829–835 (2010).
35. Uchida, H. et al. Triterpenoid levels are reduced during Euphorbia tirucalli L. callus formation. Plant Biotechnol. 27, 105–109
(2010).
36. Simo, C. C. F. et al. Benjaminamide: A new ceramide and other compounds from the twigs of Ficus benjamina (Moraceae).
Biochem. Syst. Ecol. 36, 238–243 (2009).
37. Falodun, A., Chaudhry, A. M. A. & Choudhary, I. M. Phytotoxic and chemical investigations of a Nigerian medicinal plant. Res. J.
Phytochem. 3, 13–17 (2009).
38. Wang, X., Li, C., Shi, Y. & Di, D. Two new secoiridoid glycosides from the leaves of Olea europaea L. J. Asian Nat. Prod. Res. 11,
940–944 (2009).
39. Ragasa, C. Y., Lapina, M. C., Lee, J. J., Mandia, E. H. & Rideout, J. A. Secondary metabolites from Tectona philippinensis. Nat. Prod.
Res. 22, 820–824 (2008).
40. Mbouangouere, R. N. et al. Pipthadenol A-C and α-glucosidase inhibitor from Piptadenia Africana. Res. J. Phytochem. 2, 27–34
(2008).
41. Dias, M. O., Hamerski, L. & Pinto, A. C. Semi-preparative separation of α- and β-amyrin by high performance liquid
chromatography. Quím. Nova 34, 704–709 (2011).
42. Lin, K.-W. et al. Xanthine oxidase inhibitory triterpenoid and phloroglucinol from guttiferaceous plants inhibit growth and induce
apoptosis in human NTB1 cells through a ROS-dependent mechanism. J. Agric. Food Chem. 59, 407–414 (2011).
43. Sultan, S. M. I. et al. Fungal transformation of cedryl acetate and α-glucosidase inhibition assay, quantum mechanical calculations
and molecular docking studies of its metabolites. Eur. J. Med. Chem. 62, 764–770 (2013).
44. Sua, Y., Hsu, B., Wang, E. & Ho, C. Composition, anticancer, and antimicrobial activities in vitro of the heartwood essential oil of
Cunninghamia lanceolata var. konishii from Taiwan. Nat. Prod. Commun. 7, 1245–1247 (2012).
45. Kallassy, H. et al. Chemical composition, antioxidant, anti-inammatory, and antiproliferative activities of Crataegus azarolus L.
Med. Sci. Monit. Basic Res. 23, 270–284 (2017).
46. Nisar, S. et al. Natural products as chemo-radiation therapy sensitizers in cancers. Biomed. Pharmacother. 154, 113610. h t t p s : / / d o i
. o r g / 1 0 . 1 0 1 6 / j . b i o p h a . 2 0 2 2 . 1 1 3 6 1 0 (2022).
47. Somwong, P. & eanphong, O. Quantitative analysis of triterpene lupeol and anti-inammatory potential of the extracts of
traditional pain-relieving medicinal plants Derris scandens, Albizia procera, and Diospyros rhodocalyx. J. Adv. Pharm. Technol. Res.
12, 147–151 (2021).
48. Vaidya, V. V., Pradhan, P. M. & Kondalkar, P. Simultaneous quantication of lupeol, β-sitosterol and oleanolic acid using validated
HPTLC method from Nyctanthes arbor-tristis and its marketed formulation. World J. Pharm. Res. 6, 1178–1187 (2017).
49. Roy, S., Rao, S., Bhuvaneswari, C., Giri, A. & Mangamoori, L. N. Phytochemical analysis of Andrographis paniculata extract and its
antimicrobial activity. World J. Microbiol. Biotechnol. 26, 85–91 (2010).
50. Nopsiri, W. et al. Chemical constituents and antibacterial activity of volatile oils of Combretum latifolium Bl. and C. quadrang ulare
Kurz leaves. Chiang Mai Univ. J. Nat. Sci. 14, 245–256 (2015).
51. Tram, U., Ferree, P. M. & Sullivan, W. Identication of Wolbachia–host interacting factors through cytological analysis. Microbes
Infect. 5, 999–1011 (2003).
52. Al-Reza, S. M., Rahman, A., Sattar, M. A., Rahman, M. O. & Fida, H. M. Essential oil composition and antioxidant activities of
Curcuma aromatica Salisb. Food Chem. Toxicol. 48, 1757–1760 (2010).
53. Wei, F. H., Chen, F. L. & Tan, X. M. Gas chromatographic-mass spectrometric analysis of essential oil of Jasminum ocinale L. var
grandiorum ower. Trop. J. Pharm. Res. 14, 23–26 (2015).
54. El-Hawary, S. A., Sokkar, N. M., Ali, Z. Y. & Yehia, M. M. A prole of bioactive compounds of Rumex vesicarius L. J. Food Sci. 76,
C1195–C1202 (2011).
55. Sandeep, A., Kumar, S., Singh, S. & Gupta, R. Ethnopharmacological review of medicinal plants used for wound healing in
traditional Indian medicine. J. Appl. Pharm. Sci. 8, 117–121 (2018).
56. Moy, J., Hu, W. & Hwang, P. Natural product-derived small molecules as modulators of neuroinammation. Molecules 21, 987
(2016).
57. Gautam, R., Khatri, M. & Mishra, S. Evaluation of the pharmacological properties of Solanum species: A systematic review.
Phytomedicine 24, 36–45 (2017).
58. Bhuvaneshwaran, J., Prasad, R. & Verma, A. Neuroprotective and anti-inammatory eects of Solanum species. Eur. J. Pharmacol.
763, 320–327 (2015).
59. Rani, N., Ramesh, M. & Kumar, S. Toxicological assessment of Solanum v illosum: Implications for traditional use. Environ. Toxicol.
Pharmacol. 68, 104–110 (2019).
60. Subramanian, M., Kumar, A. & irunavukkarasu, M. Phytochemical analysis and therapeutic properties of Solanum villosum.
Phytother. Res. 30, 1101–1107 (2016).
61. Sharma, R., Kumar, M. & Yadav, A. Pharmacological and medicinal properties of Solanum villosum. Int. J. Mol. Sci. 16, 5761–5773
(2015).
62. Prasad, B., Shukla, S. & Sharma, S. A review on pharmacological activities of Solanum species. J. Ethnopharmacol. 208, 83–91
(2017).
63. Reddy, P., Rao, D. & Karthik, R. Toxicological eects of Solanum villosum on liver function. Toxicol. Rep. 5, 1162–1169 (2018).
64. Das, D., Kumar, P. & Yadav, R. Phytochemical and antioxidant properties of Solanum villosum. J. Agric. Food Chem. 67, 3107–3113
(2019).
65. Shah, P., Patel, K. & akur, S. Anti-cancer potential of Solanum species: A review of pharmacological studies. Asian Pac. J. Cancer
Prev. 18, 2489–2493 (2017).
66. Tripathi, P., Kumar, A. & Meena, R. Pharmacological evaluation of Solanum species in cancer treatment. J. Pharmacol. Exp. er.
364, 285–294 (2018).
67. Borse, S., Singh, R. & Gupta, D. Exploring the therapeutic potential of Solanum species in chronic diseases. Molecules 24, 745
(2019).
68. Anwar, F., Malik, A. & Zia, M. Medicinal properties of Solanum species in folk medicine. Biol. Pharm. Bull. 40, 469–476 (2017).
69. Ali, S., Yousuf, M. & Khan, R. Solanum species: A comprehensive review on its pharmacological and toxicological prole. J. Funct.
Foods 64, 103684 (2020).
70. Chavan, R., Jain, A. & Shah, M. Phytochemistry and pharmacological properties of Solanum species. Nat. Prod. Res. 32, 1421–1425
(2018).
71. Sharma, S., Singh, J. & Verma, A. Bioactive compounds from Solanum species and their role in the treatment of chronic diseases.
Phytochem. Lett. 17, 29–35 (2016).
72. Kumar, R., Sharma, S. & Singh, B. Pharmacokinetics and pharmacodynamics of compounds from Solanum species. J. Med. Chem.
60, 2429–2435 (2017).
73. Venkatesan, R., Srinivasan, R. & Arumugam, S. Medicinal chemistry of Solanum species: A review. Eur. J. Med. Chem. 163, 77–88
(2019).
Scientic Reports | (2025) 15:4192 13
| https://doi.org/10.1038/s41598-025-87654-9
www.nature.com/scientificreports/
Content courtesy of Springer Nature, terms of use apply. Rights reserved

Acknowledgements
e authors extend their appreciation to the Researchers Supporting Project number (RSP2025R347), King Saud
University, Riyadh, Saudi Arabia, for nancial support.
Author contributions
Y.B., M.A. and M.S.A. designed the study. I.R. performed the experiments. I.R. helped in data curation and
analysis of data. M.H.S. and Y.Z collected literature reviews, helped in writing the original dra of the article
and helped in funding acquisition. Y.B. and A.Q. provided technical expertise to improve the article. All authors
reviewed and edited the manuscript.
Declarations
Competing interests
e authors declare no competing interests.
Additional information
Correspondence and requests for materials should be addressed to Y.B. or A.Q.
Reprints and permissions information is available at www.nature.com/reprints.
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and
institutional aliations.
Open Access is article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives
4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in
any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide
a link to the Creative Commons licence, and indicate if you modied the licensed material. You do not have
permission under this licence to share adapted material derived from this article or parts of it. e images or
other third party material in this article are included in the article’s Creative Commons licence, unless indicated
otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence
and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to
obtain permission directly from the copyright holder. To view a copy of this licence, visit h t t p : / / c r e a t i v e c o m m o
n s . o r g / l i c e n s e s / b y - n c - n d / 4 . 0 / .
© e Author(s) 2025
Scientic Reports | (2025) 15:4192 14
| https://doi.org/10.1038/s41598-025-87654-9
www.nature.com/scientificreports/
Content courtesy of Springer Nature, terms of use apply. Rights reserved

1.
2.
3.
4.
5.
6.
Terms and Conditions
Springer Nature journal content, brought to you courtesy of Springer Nature Customer Service Center GmbH (“Springer Nature”).
Springer Nature supports a reasonable amount of sharing of research papers by authors, subscribers and authorised users (“Users”), for small-
scale personal, non-commercial use provided that all copyright, trade and service marks and other proprietary notices are maintained. By
accessing, sharing, receiving or otherwise using the Springer Nature journal content you agree to these terms of use (“Terms”). For these
purposes, Springer Nature considers academic use (by researchers and students) to be non-commercial.
These Terms are supplementary and will apply in addition to any applicable website terms and conditions, a relevant site licence or a personal
subscription. These Terms will prevail over any conflict or ambiguity with regards to the relevant terms, a site licence or a personal subscription
(to the extent of the conflict or ambiguity only). For Creative Commons-licensed articles, the terms of the Creative Commons license used will
apply.
We collect and use personal data to provide access to the Springer Nature journal content. We may also use these personal data internally within
ResearchGate and Springer Nature and as agreed share it, in an anonymised way, for purposes of tracking, analysis and reporting. We will not
otherwise disclose your personal data outside the ResearchGate or the Springer Nature group of companies unless we have your permission as
detailed in the Privacy Policy.
While Users may use the Springer Nature journal content for small scale, personal non-commercial use, it is important to note that Users may
not:
use such content for the purpose of providing other users with access on a regular or large scale basis or as a means to circumvent access
control;
use such content where to do so would be considered a criminal or statutory offence in any jurisdiction, or gives rise to civil liability, or is
otherwise unlawful;
falsely or misleadingly imply or suggest endorsement, approval , sponsorship, or association unless explicitly agreed to by Springer Nature in
writing;
use bots or other automated methods to access the content or redirect messages
override any security feature or exclusionary protocol; or
share the content in order to create substitute for Springer Nature products or services or a systematic database of Springer Nature journal
content.
In line with the restriction against commercial use, Springer Nature does not permit the creation of a product or service that creates revenue,
royalties, rent or income from our content or its inclusion as part of a paid for service or for other commercial gain. Springer Nature journal
content cannot be used for inter-library loans and librarians may not upload Springer Nature journal content on a large scale into their, or any
other, institutional repository.
These terms of use are reviewed regularly and may be amended at any time. Springer Nature is not obligated to publish any information or
content on this website and may remove it or features or functionality at our sole discretion, at any time with or without notice. Springer Nature
may revoke this licence to you at any time and remove access to any copies of the Springer Nature journal content which have been saved.
To the fullest extent permitted by law, Springer Nature makes no warranties, representations or guarantees to Users, either express or implied
with respect to the Springer nature journal content and all parties disclaim and waive any implied warranties or warranties imposed by law,
including merchantability or fitness for any particular purpose.
Please note that these rights do not automatically extend to content, data or other material published by Springer Nature that may be licensed
from third parties.
If you would like to use or distribute our Springer Nature journal content to a wider audience or on a regular basis or in any other manner not
expressly permitted by these Terms, please contact Springer Nature at
onlineservice@springernature.com