Phytoconstituents from the Aerial Parts of Salvia dracocephaloides Boiss. and their Biological Activities

Journal of Environmental Treatment Techniques

2020, Volume 8, Issue 4, Pages: 1274-1278

J. Environ. Treat. Tech.

ISSN: 2309-1185

Journal web link: http://www.jett.dormaj.com

https://doi.org/10.47277/JETT/8(4)1278

Phytoconstituents from the Aerial Parts of Salvia

dracocephaloides Boiss. and their Biological

Activities

Salar Hafez Ghoran , Omidreza Firuzi , Amir Reza Jassbi *

Department of Chemistry, Faculty of Basic Sciences, Golestan University, Gorgan 4913815759, Iran

Medicinal and Natural Products Chemistry Research Center, Shiraz University of Medical Sciences, Shiraz 7134853734, Iran

Received: 11/07/2020 Accepted: 04/09/2020 Published: 20/12/2020

Abstract

MTT colorimetric cytotoxic bioassay, solvent fractionation and chromatographic purification of an 80% methanol extract of Salvia

dracocephaloides, led to isolation and identification of eight compounds, including 5-hydroxy-3,6,7,3ʹ,4ʹ-pentamethoxyflavone (1),

cynaroside (2), salvigenin (3), eupatorin (4), cirsimaritin (5), β-sitosterol (6), oleanolic acid (7), and ursolic acid (8). The structures of

the compounds were characterized using spectroscopic analyses including HRESI-MS, H and C-NMR and comparison with those

previously reported in the literature. The biological activities of 1 and 2, including their cytotoxicity against MCF-7 human breast cancer

cell line and DPPH free radicals scavenging effect were studied in vitro. Compound 1 showed stronger cytotoxicity; IC50 of 7.2 µg/mL

compared to that obtained for 2; IC50 of 15.5 µg/mL. While, compound 2, with 3ʹ,4ʹ dihydroquinol functionality, displayed higher DPPH

radical inhibitory activity with an IC50 of 10.7 µg/mL compared to that measured for 1; 48.8 µg/mL.

Keywords: Salvia dracocephaloides, Lamiaceae, Cytotoxicity, DPPH radical scavenging activity, Polymethoxyflavones

Introduction¹

cymene, linalool, and the sesquiterpenes; β-caryophyllene and

spathulenol [10]. Except the antioxidant potential, expressed as

DPPH radical scavenging activity of aqueous methanol extract

of S. dracocephaloides, there were no other report on the

chemical properties of the plant in the literature [9].

Hence, the main objective of the present study is the

isolation and structure elucidation of the phytochemicals

followed by reporting the cytotoxic activities of various

extracts and the unexplored isolated phytochemicals. To the

best of our knowledge this is the first report of Salvia

dracocephaloides compounds including 5-hydroxy-3,6,7,3ʹ,4ʹ-

pentamethoxyflavone; 1 [11], cynaroside; 2 [12], salvigenin; 3

Salvia L. (Sage) is one of the greatest genus of the perennial

and annual plants among the family Lamiaceae and distributed

around the world. The genus Salvia or “Maryam-Goli” in

Persian includes 62-reported species in Iran which 17 of them

are endemic and wildly grown in different parts of Iran [1, 2,

3]. Sages are rich in various specialized metabolites including

terpenoids, sterols, phenolics including flavonoids, lignans, and

phenylpropanoids [4, 5]. Salvia species are used in the folk

medicine in different parts of the world. For instance, S.

miltiorrihza; red sage, Danshen, S. officinalis; sage, and S.

sclarea; clary sage are the most common species worldwide

and S. hydrangea, S. macrosiphon, and S. mirzayanii are the

Iranian ehtnopharmacologically important ones [6]. In addition

to their use in folk medicine sage plants produce metabolites

with various biological activities such as antioxidant,

[

13], eupatorin; 4 [14], cirsimaritin; 5 [13], β-sitosterol; 6 [15],

oleanolic acid; 7, and ursolic acid; 8 [16] in the cytotoxic

chloroform soluble fraction of the 80% methanol extract

(Figure 1).

antibacterial,

antiviral,

antifungal,

antileishmanial,

antimalarial, antispasmodic, anti-inflammatory, anti-diabetic,

cytotoxic, and insecticidal activities [4, 5, 7].

2 Materials and Methods

2.1. General Experimental Procedures

As our ongoing research on the Iranian sages for their

anticancer metabolites, we have subjected S. dracocephaloides

to phytochemical analyses. S. dracocephaloides known as

All organic solvents including n-hexane, dichloromethane

(DMC), chloroform (CHCl ), ethyl acetate (EtOAc), acetone,

n-butanol (n-BuOH), methanol (MeOH), were commercially

purchased and applied without any further purification. TLC

sheets (20 × 20 cm; pre-coated silica gel 60 F254; Merck,

Germany) along with silica gel (70-230 and 230-400 mesh;

Merck, Germany) were used for analytical TLC and column

chromatography, respectively. TLC spots were detected using

anisaldehyde/sulphuric acid reagent followed by heating. HR-

ESIMS was recorded on MicrOTOF-Q II mass spectrometer

equipped with an ESI source (Bruker Daltonik, Bremen,

Germany).

“Maryam-Goli Tamashaei” in Persian is a shrubby perennial

herb 25 to 40 cm tall, with small purple flowers which grows

in rocky and mountainous regions of Caucasus and Iran [2].

The synonymous relationship between S. hydrangea and S.

dracocephaloides has been ruled out by their phenotypic

differences and various essential oils composition [2, 8]. The

major constituents of the oil of S. dracocephaloides are

camphor, 1,8-cineole, and camphene [9], while those detected

in the oil of S. hydrangea were detected as α- and β-pinene, p-

*Corresponding author: Amir Reza Jassbi, Medicinal and Natural Products Chemistry Research Center, Shiraz University of Medical

Sciences, Shiraz 7134853734, Iran. E-mail address: jassbiar@sums.ac.ir. Tel: +98 71 32303872 and Fax: +98 71 32332225

274

Journal of Environmental Treatment Techniques

2020, Volume 8, Issue 4, Pages: 1274-1278

Figure 1: Chemical structures of isolated compounds, 1–8, from S. dracocephaloides Boiss

D and 2D NMR spectra were performed on a Bruker

chromatography (FCC; 230-400 mesh) eluting with a mixture

of n-hexane/EtOAc (90:10), which resulted in six sub-

Biospin GmbH spectrometer ( H at 300 MHz and C at 75

MHz, H at 400 MHz and C at 100 MHz and H at 500 MHz

fractions; SFr. B

from MeOH in SFr. B

purified by silica gel-FCC eluting with CHCl

to yield nine sub-fractions: SFr. D -D . SFr. D -D

and re-chromatographed over silica gel FCC using the mobile

phase of CHCl :Et O; 85:15 to 70:30 to afford compound 7

-B

. Compound 6 (43.1 mg) was crystalized

while Fr. D (336.7 mg) was further

:acetone (95:5)

were pooled

and C at 125 MHz). All NMR chemical shifts were expressed

in ppm related to TMS signal at 0.00 ppm as internal reference.

.2 Plant Materials

The aerial parts of Salvia dracocephaloides Boiss. were

collected in the flowering season (June 2017) from Marand to

Jolfa road (GPS, 38˚44´N/45˚36´E, at an altitude of 1300 m),

East Azerbaijan Province, Iran and botanically characterized by

Mr. Mehdi Zare and Dr. Mojtaba Assadollahi, the plant

taxonomists in Medicinal and Natural Products Chemistry

Research Center (MNCRC). A voucher specimen; No. PC-96-

(39.4 mg). Fr. E (407.6 mg) was triturated using acetone, which

is caused to the formation of a white precipitate; compound 8

(61.7 mg). The UV quenching spots on TLC plate were

detected for the flavonoids compounds in Fr. F-H. Therefore,

Fr. F (279.0 mg) was fractionated by FCC silica gel, and eluted

with DCM/acetone (90:10) to give six sub-fractions (SFr. F -

). Compound 1 was purified as a yellow powder (10.3 mg)

upon triturating SFr. F with EtOAc. On the other hand, Fr. G

(173.5 mg) was subjected to a silica gel FCC and eluted with

CHCl /MeOH by increasing the polarity to give five sub-

fractions. The third sub-fraction was subjected to CC and

isocratically washed with CHCl /MeOH yielding the

-8-28.1, of the plant was deposited at the Herbarium MNCRC.

.3 Extraction procedure

The air-dried powder of S. dracocephaloides aerial parts

(300 g) was extracted in methanol water (8:2) solvent using

maceration method for 48 h (3 × 2L). After evaporation of the

solvent by rotary evaporator, the crude extract (71.68 g) was

partitioned in water (1L) and organic solvents (3 × 1L) in the

order of increasing polarity; n-hexane, chloroform, ethyl

acetate, and n-butanol. All of the organic layers and the

remaining water fraction were concentrated under reduced

pressure and stored in refrigerator at -20 ˚C for further

phytochemical and biological studies.

compound 3 (4.8 mg). Compounds 4 (7.6 mg) and 5 (3.9 mg)

were purified from Fr. H using silica gel-preparative thin layer

chromatography (PTLC). Eventually, Fr. K (419.3 mg) was

fractionated by silica gel FCC, eluting with gradient of

DCM/MeOH; 95;5 to 60:40), resulting eight sub-fractions

1 8

(SFr. K -K ). Amongst, the yellowish sediments of compound

2 (41.6 mg) were yielded from the SFr. K6,7 using EtOAc

trituration, washing, and filtration procedures.

.4 Purification of the phytochemicals

The CHCl and n-hexane fractions showed the highest

2.5 Cytotoxic activity

cytotoxic activity against MOLT-4 cell line (Table 1).

Therefore, the former fraction was chosen for further

purification of its constituents, while the latter one was found

to be out of interest due to the presence of high levels of fatty

acids, waxes, carotenoids, and chlorophylls detected on TLC

The in vitro cytotoxic activity of the extracts and purified

compounds (1 and 2) were examined against MOLT-4 human

lymphoblastic leukemia and MCF-7 human breast

adenocarcinoma cell lines, respectively, using 3-(4,5-

dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)

reduction assay [6, 17]. MCF-7 cells were obtained from

Iranian Biological Resource Center, Tehran, Iran, while

MOLT-4 cells were purchased from National Cell Bank of Iran,

Pasteur Institute, Tehran, Iran. The cells were seeded into 96-

well microplates and incubated overnight at 37 °C. Three to

four different concentrations of plant extracts or isolated

compounds were added in triplicate and incubated for another

72 h. The concentration of DMSO in each well did not exceed

0.25%. At the end of the incubation time, the media in each

well was replaced with 0.5 mg/mL MTT dissolved in RPMI

without phenol red. After 4 hours at 37 °C DMSO was added

to solubilize the formazan crystals formed inside the viable

sheet. An aliquot of the CHCl fraction (5 g) was subjected to

silica gel open column chromatography (CC; 35 × 5 cm; 250

g), eluted with n-hexane followed by increasing the polarity to

pure EtOAc and then to EtOAc:MeOH (50:50, v/v). The

resulting CC fractions were analyzed by TLC using different

mobile phase (n-Hex:EtOAc 3:7 and EtOAc:MeOH 9:1) and

visualizing with anisaldehyde/sulphuric acid reagent followed

by heating on a hot plate. The similar CCʹs fractions on TLC

were mixed to afford 12 fractions (Fr. A-L). The well separated

fractions on analytical TLCs were selected for further

purification using preparative chromatography. Thus, Fr. B

(290.1 mg) was loaded onto silica gel flash column

275

Journal of Environmental Treatment Techniques

2020, Volume 8, Issue 4, Pages: 1274-1278

cells. The optical density of the final solution was measured at

antioxidant activity [18, 19, 20], while the former (1) has been

isolated from the leaves of Vitex negundo acting as anti-allergic

constituent [21]. Most of the Polymethoxyflavones (PMFs)

have been reported form the Citrus species, from leaves of

Murraya spp. (Rutaceae), Kaempferia spp. (Zingiberaceae),

and Piper spp. (Piperaceae) [22]. Despite the presence of

5,6,7,8-tetra-oxygeneted flavones among the plants of

Lamiaceae family only a few species including Thymus, Salvia,

Vitex, and Ocimum appear to accumulate ploymethoxylated

flavones [21, 23, 24].

70 nm using a Bio-Tek microplate reader (Model Synergy

HTX). The cell viability was calculated compared to untreated

control cells by comparison of absorbance measurements. IC50

values for each compound was calculated using CurveExpert

software version 1.34 for Windows. Each experiment was

repeated 3 to 5 times. Paclitaxel, a standard chemotherapeutic

agent, was used as a positive control.

.6 DPPH radical scavenging activity

The in vitro antioxidant activity of compounds 1 and 2 was

According to the literature survey, Walle reported that the

most cytotoxic active flavones are those ones bearing the

methoxy groups and concluded that the glycosylation is a

crucial effect resulting to be inactive agent on the biological

activity [25]. The cytotoxicity against MCF-7 cell lines of

compound 1 and quercetin are the highest among the tested

compounds followed by compound 2, respectively (Table 1).

This is in agreement with the extensive research on the

cytotoxicity of PMFs. The higher activity is attributed to the

presence of hydroxy group at position C-5 of the PMFs, since

5-hydroxy PMFs exhibited stronger anticancer activity when

tested against the colon cancer cells compared to the respective

5-methoxylated derivatives [26]. Salvigenin, eupatorin, and

cirsimaritin were isolated from Centaurea kilaea (Asteraceae)

in a bioassay-guided purification procedure against two

determined calorimetrically by assaying for DPPH (2,2-

diphenyl-1-picrylhydrazyl) radical scavenging activity [7].

Briefly, the plant extracts or isolated compounds were diluted

in methanol at five concentrations (78-1250 μg.mL ). Five

microliters of each concentration were added to 195 μL of

DPPH solution (100 μM concentration in MeOH). The

microplate was shaken at room temperature in the dark place.

After 30 min, the absorbance was measured at 517 nm using a

Bio-Tek microplate reader (Model Synergy HTX). IC50 values

for each components was calculated using CurveExpert

software version 1.34 for Windows. Each experiment was

repeated 3 times. Quercetin, a pentahydroxy-flavonol, was used

as the positive control.

cancerous cell lines; MCF-7 and PC-3;

prostate

Results and Discussion

adenocarcinoma cell line. Only the two later compounds

showed potent cytotoxic effect against PC-3 [27, 28]. Quercetin

alone or in combination with topotecan showed significant

cytotoxicity against the MCF-7 cell lines [29]. While

cynaroside showed weak cytotoxic effect against the breast

cancer cell line [30]. Literature survey confirmed that

polymethoxylated flavonoids like nobiletin, tangeretin,

sinensetin, and 5-demethyltangeretin (found in Citrus spp.)

play a key role in anticancer and anti-inflammatory activity [21,

Methanol-water (80:20) extract of the aerial parts of S.

dracocephaloides after evaporation of its methanol was

partitioned between solvents with different polarities; n-

hexane, chloroform, ethyl acetate, and n-butanol. The resulting

fractions were evaluated for their cytotoxic effects against

MOLT-4 human leukemia cell line as well as DPPH radical

scavenging activity. The active chloroform layer was subjected

to column chromatography over normal phase silica gel.

Consequently, eight reported compounds (1-8) were isolated

and characterized (Figure 1). The chemical structures of the

isolated compounds 3-8 were elucidated based on comparison

of their physical constants such as co-TLC with the reference

substances that has been simultaneously isolated from S.

russellii in our laboratory and their structures were elucidated

by H and C NMR Spectroscopy. Meanwhile, the chemical

structures of 1 and 2 were identified as 5-hydroxy-3,6,7,3ʹ,4ʹ-

pentamethoxyflavone and cynaroside (luteolin-7-O-β-D-

glucopyranoside) based on the H and C NMR spectral data,

respectively [11, 12]. The later compound showed a vast

variety of biological activities such as antimalarial,

2, 31], as we found for compound 1. In addition, the cytotoxic

activities of compounds 6-8 have also been previously

examined by other investigators against MCF-7 cell line [32,

3, 34]. The radical scavenging activity of quercetin is higher

than those measured for compound 1 and 2 as the following

descending order of IC50s: quercetin (5.3 µg.mL )

‒

cynaroside;

(10.7 µg.mL )

5-hydroxy-3,6,7,3ʹ,4ʹ-

‒

pentamethoxyflavone; 1 (48.8 µg.mL ). Previously, it was

reported that glycosylation or methylation of the hydroxy

groups decreases the antioxidant activity of flavonoids while

the free hydroxy groups together with 3ʹ,4ʹ dihydroquinol

functionality increase it dramatically (supplementary data) [35,

antileishmanial,

cytotoxicity,

anti-inflammatory

and

6].

Table 1: Cytotoxic and antioxidant activities of various fractions and pure flavonoids isolated from S. dracocephaloides Boiss.

Antioxidant activity IC50

(µg/mL)

Cytotoxic activity IC50 (µg/mL)^a

Weight of the

extract (g)

Materials

•

MOLT-4 cells

MCF-7 cells

DPPH radicals

Aqueous methanol extract

n-Hexane fraction

73.68

15.35

7.26

6.73

14.32

25.61

35.5 ± 12.2

24.6 ± 8.0

25.8 ± 6.5

31.7 ± 6.9

35.3 ± 16.6

NA^b

47.36 ± 1.89

35.76 ± 4.29

57.91 ± 2.81

76.63 ± 5.62

CHCl

fraction

EtOAc fraction

n-BuOH fraction

Aqueous fraction

-hydroxy-3,6,7,3ʹ,4ʹ-

NT^c

7.22 ± 2.95

48.82 ± 3.43

pentamethoxy flavone (1)

Cynaroside (2)

Quercetin (µg/ml)

Paclitaxel (ng/ml)

2.4 ± 0.7

15.48 ± 2.04

7.16 ± 1.35

10.74 ± 2.78

5.29 ± 2.36

2.49 ± 0.57

The data are presented as mean ± S.D. of at least three independent experiments. NA: Not active. NT: Not tested. Paclitaxel and quercetin were tested

as reference agents.

276

Journal of Environmental Treatment Techniques

2020, Volume 8, Issue 4, Pages: 1274-1278

0. Rustaiyan A, Masoudi S, Jassbi AR. Essential Oil of Salvia

hydrangea DC. ex Benth. Journal of Essential Oil Research.

Conclusion

To the best of our knowledge, this is the first report of non-

997;9(5):599-600.

volatile phytochemicals in S. dracocephaloides resulting in

identification of one PMF (1), further four flavones (2-5), β-

sitosterol and two triterpenoids (6-8). Compound 1 present a

rare flavone in Salvia species with five methoxy groups, so far

reported in the plants of the Lamiaceae family. Phytochemical

investigations on Salvia species revealed that the natural

occurrence of 3-methoxy flavones (flavonol-3-methyl-ether) is

rare with the expectation of a few compounds including,

isokaempferide, kumatakenin, quercetin 3-methyl ether,

ayanin, and retusin previously reported [37]. Taken together,

the polymethoxyflavones and flavonols could be considered as

the chemotaxonomic markers for S. dracocephaloides".

Likewise, compound 1 acted as a potent cytotoxic agent while

weakly scavenged the free stable radicals of DPPH that showed

the activities are not follow the same mechanisms of action.

1. Martínez V, Barberá O, Sanchez-Parareda J, Marco JA. Phenolic

and acetylenic metabolites from Artemisia assoana.

Phytochemistry. 1987;26(9):2619-2624.

12. Blunder M, Orthaber A, Bauer R, Bucar F, Kunert O. Efficient

identification of flavones, flavanones and their glycosides in

routine analysis via off-line combination of sensitive NMR and

HPLC experiments. Food Chemistry. 2017;218:600-609.

3. Alwahsh MA, Khairuddean M, Chong WK. Chemical constituents

and antioxidant activity of Teucrium barbeyanum Aschers.

Records of Natural Products. 2015;9(1):159-163.

4. Nagao T, Abe F, Kinjo J, Okabe H. Antiproliferative constituents

in plants 10. Flavones from the leaves of Lantana montevidensis B

RIQ. and consideration of structure–activity relationship.

Biological and Pharmaceutical Bulletin. 2002;25(7):875-879.

5. Saeidnia S, Manayi A, Gohari AR, Abdollahi M. The story of beta-

sitosterol-a review. European Journal of Medicinal Plants.

014;4(5):590-609.

6. Martins D, Carrion LL, Ramos DF, Salomé KS, da Silva PE,

Barison A, Nunez CV. Triterpenes and the antimycobacterial

activity of Duroia macrophylla Huber (Rubiaceae). BioMed

Research International. 2013 Oct;2013.

7. Hafez Ghoran S, Babaei E, Rezaei Seresht H, Karimzadeh Z.

Cytotoxic constituents and molecular docking study of the active

triterpenoids from Tripleurospermum disciforme (C. A. Mey.)

Schultz-Bip. Jundishapur Journal of Natural Pharmaceutical

Products. 2020;15(2):e65760.

8. Calixto JB, Campos MM, Otuki MF, Santos AR. Anti-

inflammatory compounds of plant origin. Part II. Modulation of

pro-inflammatory cytokines, chemokines and adhesion molecules.

Planta Medica. 2004;70(2):93-103.

9. Kirmizibekmez H, Calis I, Perozzo R, Brun R, Donmez AA,

Linden A, Rudi P, Tasdemir D. Inhibiting activities of the

secondary metabolites of Phlomis brunneogaleata against

parasitic protozoa and plasmodial enoyl-ACP reductase, a crucial

enzyme in fatty acid biosynthesis. Planta Medica. 2004;70(8):711-

717.

Acknowledgments

The authors are grateful to Golestan University and Shiraz

University of Medical Sciences for the financial support. We

especially thankful to Dr. Mojtaba Asadollahi and Mr. Mehdi

Zare for their skillful collection and identification of the plant

material. This article is a part of the PhD dissertation submitted

to the Department of Chemistry (Golestan University, Gorgan,

Iran), for the fulfillment of the PhD degree of Salar Hafez

Ghoran in Phytochemistry.

Authorship contribution statement

Conceptualization,

Investigation,

Methodology,

Roles/Writing – original draft: S. Hafez Ghoran. Conducted

experiments: S. Hafez Ghoran, A.R. Jassbi. Writing - review &

editing, Validation, Supervision: O. Firuzi, A.R. Jassbi. All the

authors approved the final version of the manuscript.

0. Saracoglu I, Varel M, Harput US, Nagatsu A. Acylated flavonoids

and phenol glycosides from Veronica thymoides subsp.

pseudocinerea. Phytochemistry. 2004;65(16):2379-2385.

1. Patel J, Deshpande SS. Anti-Allergic and antioxidant activity of 5-

Hydroxy-3,6,7,3ʹ,4ʹ-pentamethoxy flavone isolated from leaves of

Vitex negundo. Anti-Inflammatory & Anti-Allergy Agents in

Medicinal Chemistry (Formerly Current Medicinal Chemistry-

Anti-Inflammatory and Anti-Allergy Agents). 2011;10(6):442-451.

Conflict of interest

The authors declare they have no any competing interest.

References

Hedge I. Salvia L. Flora of Turkey and the East Aegean islands.

982;7:400-461.

Jamzad Z, Asadī M. ''Lamiaceae. Research Institute of Forests and

Rangelands; 2012.

Mozaffarian V. A dictionary of Iranian plant names: Latin,

English, Persian. Farhang Mo'aser; 2003.

Jassbi AR, Zare S, Firuzi O, Xiao J. Bioactive phytochemicals

from shoots and roots of Salvia species. Phytochemistry reviews.

22. Tung YC, Chou YC, Hung WL, Cheng AC, Yu RC, Ho CT, Pan

MH. Polymethoxyflavones: Chemistry and molecular mechanisms

for cancer prevention and treatment. Current Pharmacology

Reports. 2019;5(2):98-113.

23. Berim A, Gang DR. Methoxylated flavones: occurrence,

importance,

biosynthesis.

Phytochemistry

Reviews.

016;15(5):829-867.

2016;15(3):363-390.

Wu YB, Ni ZY, Shi QW, Dong M, Kiyota H, Gu YC, Cong B.

Constituents from Salvia species and their biological activities.

Chemical Reviews. 2012;112(11):5967-6026.

24. Tomás-Barberán F, Wollenweber E. Flavonoid aglycones from the

leaf surfaces of some Labiatae species. Plant Systematics and

Evolution. 1990;173(3-4):109-118.

Asadollahi M, Firuzi O, Jamebozorgi FH, Alizadeh M, Jassbi AR.

25. Walle T. Methoxylated flavones,

superior cancer

Ethnopharmacological

studies,

chemical

composition,

chemopreventive flavonoid subclass? Seminars in cancer biology.

2007;17(5):354-362.

26. Qiu P, Dong P, Guan H, Li S, Ho CT, Pan MH, McClements DJ,

Xiao H. Inhibitory effects of 5‐hydroxy polymethoxyflavones on

colon cancer cells. Molecular Nutrition & Food Research.

2010;54(S2):S244-S252.

27. Sen A, Ozbas Turan S, Bitis L. Bioactivity-guided isolation of anti-

proliferative compounds from endemic Centaurea kilaea.

Pharmaceutical Biology. 2017;55(1):541-546.

28. Kouhbanani MA, Beheshtkhoo N, Amani AM, Taghizadeh S,

Beigi V, Bazmandeh AZ, Khalaf N. Green synthesis of iron oxide

nanoparticles using Artemisia vulgaris leaf extract and their

application as a heterogeneous Fenton-like catalyst for the

degradation of methyl orange. Materials Research Express.

2018;5(11):115013.

antibacterial and cytotoxic activities of essential oils of eleven

Salvia in Iran. Journal of Herbal Medicine. 2019;17:100250.

Firuzi O, Miri R, Asadollahi M, Eslami S, Jassbi AR. Cytotoxic,

antioxidant and antimicrobial activities and phenolic contents of

eleven Salvia species from Iran. Iranian Journal of

Pharmaceutical Research: IJPR. 2013;12(4):801-810.

Kotan R, Kordali S, Cakir A, Kesdek M, Kaya Y, Kilic H.

Antimicrobial and insecticidal activities of essential oil isolated

from Turkish Salvia hydrangea DC. ex Benth. Biochemical

Systematics and Ecology. 2008;36(5-6):360-368.

Jamzad M, Hafez-Taghva P, Kazembakgloo A, Jamzad Z.

Chemical composition of essential oil, total flavonoid content and

antioxidant activity of Salvia dracocephaloides Boiss. from Iran.

Journal of Essential Oil Bearing Plants. 2014;17(6):1203-1210.

9. Akbas SH, Timur M, Ozben T. The Effect of Quercetin on

Topotecan Cytotoxicity in MCF-7 and MDA-MB 231 Human

277

Journal of Environmental Treatment Techniques

2020, Volume 8, Issue 4, Pages: 1274-1278

Breast Cancer Cells1. Journal of Surgical Research.

005;125(1):49-55.

0. Mamadalieva NZ, Herrmann F, El‐Readi MZ, Tahrani A, Hamoud

R, Egamberdieva DR, Azimova SS, Wink M. Flavonoids in

Scutellaria immaculata and S. ramosissima (Lamiaceae) and their

biological activity. Journal of Pharmacy and Pharmacology.

011;63(10):1346-1357.

1. Manthey JA, Guthrie N, Grohmann K. Biological properties of

Citrus flavonoids pertaining to cancer and inflammation. Current

Medicinal Chemistry. 2001;8(2):135-153.

2. Chakravarti B, Maurya R, Siddiqui JA, Bid HK, Rajendran SM,

Yadav PP, Konwar R. In vitro anti-breast cancer activity of

ethanolic extract of Wrightia tomentosa: role of pro-apoptotic

effects of oleanolic acid and urosolic acid. Journal of

Ethnopharmacology. 2012;142(1):72-79.

3. Kassi E, Sourlingas TG, Spiliotaki M, Papoutsi Z, Pratsinis H,

Aligiannis N, Moutsatsou P. Ursolic acid triggers apoptosis and

Bcl-2 downregulation in MCF-7 breast cancer cells. Cancer

investigation. 2009;27(7):723-733.

4. Awad AB, Chinnam M, Fink CS, Bradford PG. β-Sitosterol

activates Fas signaling in human breast cancer cells.

Phytomedicine. 2007;14(11):747-754.

5. Jassbi AR, Singh P, Krishna V, Gupta PK, Tahara S. Antioxidant

study and assignments of NMR spectral data for 3′,4′,7-

trihydroxyflavanone 3′,7-di-O-β-D-glucopyranoside (butrin) and

its hydrolyzed product. Chemistry of Natural Compounds.

004;40(3):250-253.

6. S Tuzun B, Hajdu Z, Orban-Gyapai O, P Zomborszki Z, Jedlinszki

N, Forgo P, Kıvcak B, Hohmann J. Isolation of chemical

constituents of Centaurea virgata lam. and xanthine oxidase

inhibitory activity of the plant extract and compounds. Medicinal

Chemistry. 2017;13(5):498-502.

7. Lu Y, Foo LY. Polyphenolics of Salvia—a review.

Phytochemistry. 2002;59(2):117-140.

278