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Rosa rugosa petals are used for production of teas, jams, wines and juices. Despite the wide availability of rose cultivars comprehensive information on petals chemical composition and healthful properties is still lacking. Therefore, the aim of this study was analysis of cytotoxic, antioxidant, antimicrobial activity and chemical composition of rugosa rose petals. Petals of R. rugosa were evaluated for their cytotoxic effect against cervical (HeLa) and breast cancer (T47D) cell lines and for antiradical activity (with DPPH(•) ). As a result, significant cytotoxic (up to 100% of dead cells) and antiradical properties (IC50 1.33 - 0.08 mg mg(-1) DPPH(•) ) were demonstrated. Moreover, notable antimicrobial activity against eight bacterial (i.e. S. epidermidis, S. aureus, B. subtilis, M. luteus, E. coli, K. pneumoniae, P. aeruginosa, P. mirabilis) and two yeast strains (C. albicans, C. parapsilosis) was showed. Total phenolic, flavonoid, phenolic acid, tannin, carotenoid and polysaccharide content in petals was determined using spectrophotometric methods. LC-ESI-MS/MS was used to thoroughly analyze phenolic acids and flavonoid glycosides in the methanolic extract and fractions obtained after its separation. Five phenolic acids and six flavonoids previously not reported in the plant material were identified. This is the first such detailed report on chemical composition and biological activity of Rcxh rugosa petals.
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Research Article
Received: 6 May 2013 Revised: 21 June 2013 Accepted article published: 1 July 2013 Published online in Wiley Online Library:
( DOI 10.1002/jsfa.6294
Cytotoxic, antioxidant, antimicrobial
properties and chemical composition
of rose petals
Renata Nowak,aMarta Olech,aŁukasz Pecio,bWiesław Oleszek,b
Renata Los,c† Anna Malmcand Jolanta Rzymowskad
BACKGROUND: Rosa rugosa petals are used for production of teas, jams, wines and juices. Despite the wide availability of rose
cultivars, comprehensive information on petal chemical composition and healthful properties is still lacking. Therefore, the aim
of this study was analysis of cytotoxic, antioxidant and antimicrobial activity and chemical composition of rugosa rose petals.
RESULTS: Petals of R. rugosa were evaluated for their cytotoxic effect against cervical (HeLa) and breast cancer (T47D) cell lines
and for antiradical activity (with DPPH). As a result, significant cytotoxic (up to 100% of dead cells) and antiradical properties
(IC50 1.33 –0.08 mg mg1DPPH) were demonstrated. Moreover, notable antimicrobial activity against eight bacterial (i.e.
Staphylococcus. epidermidis,S. aureus,Bacillus subtilis,Micrococcus luteus,Escherichia coli,Klebsiella pneumoniae,Pseudomonas
aeruginosa,Proteus mirabilis) and two yeast strains (Candida. albicans, C. parapsilosis) was shown. Total phenolic, flavonoid,
phenolic acid, tannin, carotenoid and polysaccharide content in petals was determined using spectrophotometric methods.
Liquid chromatographyelectrospray ionizationtandem mass spectrometry was used to thoroughly analyze phenolic acids
and flavonoid glycosides in the methanolic extract and fractions obtained after its separation. Five phenolic acids and six
flavonoids previously not reported in the plant material were identified.
CONCLUSION: This is the first such detailed report on chemical composition and biological activity of R. rugosa petals.
2013 Society of Chemical Industry
Keywords: Rosa rugosa; LC-MS; radical scavenging activity; antibacterial activity; cytotoxic effect; phenolics
Plant foods are indispensable components of a healthy diet
providing a wide spectrum of substances (e.g. vitamins, minerals,
fiber). Moreover, plants are increasingly appreciated because
of the chemopreventive properties of their metabolites. Plants’
chemopreventive potential depends to a large extent on
phytochemicals demonstrating antioxidant and antiproliferative
activity.1Owing to the high toxicity of synthetic compounds,
the search for new natural cytotoxic, antiradical, as well as
antimicrobial substances still remains a challenge for modern
Rugosa rose (Rosa rugosa Thunb.) belongs to rose cultivars
grown primarily for their large hips, commonly used for food
and medicinal purposes. In many countries its petals are used
for production of teas, jams, wines and juices.2This species is
particularly popular in Asian countries, where petals and roots
have also been utilized in traditional medicine for the treatment
of diarrhea, injuries, gastroenteritis, hepatitis, dysmenorrhea and
blood circulation disorders, as well as pain management and
hemostasis maintenance.35
Despite the wide availability of raw materials obtained from
R. rugosa, their utilization is relatively limited. This is likely to
result from poor knowledge about the nutritional and medicinal
properties of the species, lack of comprehensive information on
the chemical composition and biological activity, in particular.
No data about the content of the major groups of active
ingredients are available and the fraction of phenolic acids and
flavonoids are still poorly explored. Only the compositions of the
petal essential oil, hydrolysable tannin fraction and anthocyanin
fraction have been well known.6,7 In terms of biological activity, the
greatest attention was paid to antioxidant properties of different
petal extracts.2,8,9 The only information concerning anticancer
activity of petals comes from our previous screening study.10 The
study in question has also revealed an important contribution of
phenolic compounds to the biological activity of rose extracts,
Correspondence to: Renata Nowak, Department of Pharmaceutical Botany,
Medical University, 1 Chod´
zki Street, 20– 093 Lublin, Poland. E-mail:
aDepartment of Pharmaceutical Botany, Medical University, 20-093 Lublin,
bDepartment of Biochemistry and Crop Quality, Institute of Soil Science and
Plant Cultivation, State Research Institute, 24-100 Pulawy, Poland
cDepartmentof Pharmaceutical Microbiology, MedicalUniversity,20-093 Lublin,
dDepartmentof Biology and Genetics, Medical University, 20 093Lublin, Poland
J Sci Food Agric (2013) c
2013 Society of Chemical Industry R Nowak et al.
and initially demonstrated interesting antibacterial potential.
Antifungal properties have not been studied to date.
The aforementioned research of other authors and our
preliminary studies indicate that rugosa rose petals are of
significant potential health benefits.3,10 Therefore, the objective of
the present study was to thoroughly examine the composition
(especially phenolic constituents) and biological activities
(antiradical, antibacterial, antifungal and cytotoxic, in particular)
of R. rugosa petals. Moreover, an attempt was made to determine
which groups of compounds were responsible for a given type of
biological activity.
Plant material
Petals of R. rugosa were collected in Serock (near Lublin, Poland) in
June 2009. The plant specimen was authenticated by Prof. Tadeusz
Krzaczek and deposited at the Department of Pharmaceutical
Botany, Medical University of Lublin, Poland. The plant material
was dried at ambient temperature and powdered according to the
European Pharmacopoeia, 6th edition.11
Standards of gallic, protocatechuic, gentisic, caffeic, synapic,
p-coumaric and salicylic acids, pyrogallol, ascorbic acid, Trolox,
2,2-diphenyl-1-picrylhydrazyl radical (DPPH), hide powder and
quercetin were purchased from Sigma-Aldrich Fine Chemicals
(St Louis, MO, USA). Flavonoids: quercetin-3-L-arabinofuranoside
(avicularin), quercetin-3-O-rutinoside (rutin), quercetin-3-O-
galactoside (hyperoside), quercetin-3-O-glucoside (isoquercitrin),
kaempferol-3-O-(6-O-(E)-p-coumaroyl)-glucoside (tiliroside),
kaempferol-3-O-glucoside (astragalin), quercetin-3-O-rhamnoside
(quercitrin), kaempferol-3-O-rutinoside (nicotiflorin) and apigenin-
7-O-glucoside were from ChromaDex (Irvine, CA, USA). Ethanol,
methanol, chloroform, hexane, diethyl ether, ethyl acetate, sodium
molybdate, sodium nitrite, dimethyl sulfoxide (DMSO), isoamyl
alcohol and FolinCiocalteu reagent were from POCh (Gliwice,
Poland), β-carotene was from Fluka (Buchs, Switzerland), and β-D-
(+)-glucose and D-(+)-xylose were from ChromaDex (Irvine, USA).
All chemicals were of analytical grade. Liquid chromatography
(LC)-grade methanol (MeOH) and acetonitrile (ACN) were
purchased from Merck (Darmstadt, Germany). LC-grade water was
prepared using a Milli-Q purification system (Millipore, Bedford,
All spectrophotometric assays were conducted in the Thermo
Scientific Evolution 300 spectrophotometer (Lafayette, CO, USA).
Analysis of the major groups of active ingredients in plant
Total phenolic, phenolic acid, flavonoid and tannin content
The total phenolic content in plant material was determined
according to the Singleton and Rossi method, with some
modifications.12 The results were expressed as milligrams of gallic
acid per gram of dry plant material.
In order to evaluate the content of phenolic acids the method
previously described by Nichiforesco and Coucou was used.13
Phenolic acid content was calculated using a reference curve
plotted for caffeic acid and expressed as milligrams of caffeic acid
per gram of dry plant material.
Total flavonoid content in plant material was determined
colorimetrically according to the method described by Lamaison
and Carret.14 Results were expressed as milligrams of quercetin
per gram of dry plant material.
Determination of tannin content in R. rugosa petals was
performed according to the method described in the Polish
Pharmacopoeia (6th edition) with some modifications.15 Tannins
were estimated indirectly after adsorption on and precipitation
with insoluble hide powder. Results were expressed as milligrams
of pyrogallol per gram of dry plant material.
Total carotenoid content
The carotenoid content in R. rugosa petals was determined
spectrophotometrically by means of the modified method
described in Current Protocols in Food Analytical Chemistry16 and
expressed as micrograms of β-carotene (reference standard) per
gram of dry plant material.
Total polysaccharide content
To analyze total polysaccharide content a weighed amount (10 g)
of dried and pulverized plant material was macerated with 100 mL
of 95% ethanol at ambient temperature for 16 h to remove most
of the polyphenols and monosaccharides, then filtered and left
to dry in air. Subsequently, plant material was extracted twice
(for 3 h each) with water (1:20, w/v) at ambient temperature and
additionally the material was extracted with water by reflux in
a water bath at 90 C for 1 h. The extracts were combined and
concentrated to 25 mL using a rotary evaporator. The residue was
extracted with 25 mL of the Sevag reagent (2% isoamyl alcohol in
chloroform) to deproteinize. After removal of the Sevag reagent,
cold anhydrous ethanol (1:4, v/v) was added to the water phase,
and the mixture was kept overnight at 4 C to precipitate the
polysaccharides. The sediment was centrifuged and lyophilized.
Total sugar content was determined by the phenolsulfuric acid
colorimetric method, using glucose (490 nm) and xylose (480nm)
as standards.17
Liquid chromatographyelectrospray ionization–tandem
mass spectrometry (LC-ESI-MS/MS) of phenolic composition
of rose petals
Preparation of samples
Twenty-five grams of dried and pulverized rose petals were
extracted four times (for 24 h each), at ambient temperature
with 150 mL of 80% (v/v) aqueous methanol. During the last
extraction, a sonication step at 50 C (30 min) was applied in
order to achieve exhaustive extraction. Extracts were combined,
filtered and evaporated to dryness under vacuum. The residue was
weighed and redissolved in 80% (v/v) methanol at a concentration
of 100 mg mL1(M). Part of this solution (20 mL) was lyophilized
in Free Zone 1 apparatus (Labconco, Kansas City, MO, USA), then
stored in the refrigerator prior to further analysis.
The remaining amount of M was fractionated into diethyl ether,
ethyl acetate and water fractions. Briefly, 80 mL of the M solution
was evaporated to dryness under vacuum. Syrupy residue was
dissolved in hot water, left to cool (12 h, 4 C), filtered and filled
with distilled water to a volume of 100 mL. Subsequently, elution
with solvents with increasing polarity was carried out. Initially,
the aqueous fraction was extracted with six portions of diethyl
ether (50 mL). Collected ether extracts were combined, the solvent
was evaporated and the residue was lyophilized to give the ether
fraction (FE).
Aqueous solution after extraction with diethyl ether was further
washed six times with portions (50 ml) of ethyl acetate. Eluates c
2013 Society of Chemical Industry J Sci Food Agric (2013)
Biological activity and chemical composition of rose petals
were combined, concentrated in a vacuum evaporator and the
residue was lyophilized to give the acetate fraction (OE).
Aqueous solution remaining after the extractions was also
evaporated, freeze-dried and used for further analysis as the
water fraction (W).
Prior to LC-MS analysis methanolic extract and its fractions were
cleansed of ballast substances using solid-phase extraction on
octadecyl SPE columns (500 mg, JT Baker Inc., Philipsburg, NJ,
USA). All extracts were prepared in triplicate.
LC-MS/MS conditions of analysis of phenolic acids and flavonoid
Phenolic acid and flavonoid glycoside contents in methanolic
extract (M), diethyl ether (FE), ethyl acetate (OE) and water (W)
fraction were determined by reversed-phase ultra-high-pressure
LC-MS, performed on an ACQUITY UPLC System (Waters Corp.,
Milford, MA, USA). For the specific detection of analytes a Waters
DAD and TQ detector in negative electrospray ionization mode
with metastable reaction monitoring (MRM) were used. Argon
was used as collision gas (collision energies are given in Table 1)
and nitrogen as desolvation gas (500 L h1and 1000 L h1for
phenolic acids and flavonoids respectively). The data were
acquired and processed using MassLynx software (Waters Corp.).
Data acquisition was carried out using different retention time
windows. Triplicate injections were made for each standard
solution and sample. Calibration curves were obtained by plotting
the peak area of the phenolic compound at each level prepared
against the concentration of standard solutions and showed linear
relationships. Compounds were quantified using the external
standard method.
For phenolic acids, chromatographic analyses were carried out
on a Waters ACQUITY UPLCHSS T3 column (1.0 ×100 mm;
1.8 µm; Waters Corp.) utilizing a gradient elution program with
a mobile phase A (water containing 0.1% HCOOH) and a mobile
phase B (ACN). The solvent A concentration was changed as
follows: 0 min (95%); 0.8 min (90%); 2.0 min (85%); 3.6 min (79%);
5.3 min (73%); 7.6 min (50%); 8.6 min (5%); 10.0 min (95%); with
completion at 11 min. The flow rate was 0.06 mL min1and column
temperature was 30 C.
The injection volume was 3 µL. In the MRM mass range from
50 to 400 m/zwas recorded and the following parameters were
applied: capillary voltage 2.4 kV, source temperature 110 Cand
desolvation temperature 350 C.
In the case of flavonoid glycosides, separations were carried
out on a Waters ACQUITY UPLCBEH C18 (2.1 ×100 mm;
1.7 µm; Waters Corp.). A gradient elution program with a mobile
phase A (water containing 0.1% HCOOH) and a mobile phase
B (ACN containing 0.1% HCOOH) was used. The solvent A
concentration was changed as follows: 0 min (87%); 0.5 min (87%);
6.0 min (85.5%); 8.5 min (60%); 9.5 min (60%); 9.6 min (87%); with
completion at 11.6 min. The flow rate was 0.04 mL min1and
column temperature was 50 C. The injection volume was 5 µL.
The mass range from 200 to 800 m/zwas recorded and the
following parameters were applied: capillary voltage 2.8 kV, source
temperature 120 C, desolvation temperature 350 C.
The data were acquired and processed using MassLynx software
(Waters Corp.). Data acquisition was carried out using different
retention time windows. Triplicate injections were made for each
standard solution and sample. Calibration curves were obtained
by plotting the peak area of the phenolic compound at each level
prepared against the concentration of standard solutions and
showed linear relationships. Compounds were quantified using
the external standard method.
The limits of detection (LOD) and quantification (LOQ) for
phenolic compounds were determined at a signal-to-noise ratio of
Table 1. LC/ESI-MS/MS analytical results of phenolic compounds in Rosa rugosa petals
Peak tR(min)
UV max.
(nm) [M H]
Products of
energy (eV) Compound
(ng µL1)
(ng µL1)
range(ng µL1)
Phenolic acids
1 3.42 271.0 169 124.8 15 Gallic acid 0.028 0.093 0.10– 10.00
2 4.48 260.0 294.0 153 108.0 25 Protocatechuic acid 0.018 0.059 0.10– 10.00
109.0 15
3 5.65 237.0 328.0 153 109.0 15 Gentisic acid 0.017 0.057 0.10– 10.00
4 6.10 322.7 179 134.9 20 Caffeic acid 0.017 0.057 0.10– 10.30
5 7.28 225.7 308.7 163 118.9 15 p-Coumaric acid 0.018 0.061 0.10– 10.20
6 7.64 236.7 323.7 223 163.8 15 Synapic acid 0.032 0.107 0.10– 10.00
207.8 15
7 8.72 235.7 300.7 137 93.0 15 Salicylic acid 0.017 0.057 0.10–10.20
Flavonoid glycosides
1 3.00 255.7 355.7 609 300 35 Rutin 0.86 2.60 0.98– 9.65
2 3.09 255.7 353.7 463 300 25 Hyperoside 1.12 3.40 1.00– 10.00
3 3.36 256.7 356.7 463 300 25 Isoquercitrin 0.47 1.42 1.00– 10.00
4 4.41 264.7 349.7 447 284 25 Astragalin 1.13 3.43 1.00– 10.00
5 4.62 257.7 349.7 433 300 25 Avicularin 0.89 2.69 0.89 8.72
6 4.81 264.7 349.7 593 285 30 Kaempferol-3-O-rutinoside 0.44 1.34 1.00– 10.00
7 5.41 256.7 352.7 447 300 25 Quercitrin 1.07 3.23 1.00– 10.00
8 6.30 266.7 337.7 431 268 30 Apigenin-7-O-glucoside 0.69 2.10 1.00– 10.00
9 8.28 266.7 314.7 593 285 30 Tiliroside 0.86 2.61 1.00– 10.00
LOD, limit of detection; LOQ, limit of quantification values and linearity ranges for identified compounds.
Compounds were confirmed by comparison with authentic standards.
J Sci Food Agric (2013) c
2013 Society of Chemical Industry R Nowak et al.
3:1 and 10:1, respectively, by injecting a series of dilute solutions
with known concentrations.
The scavenging effect of samples was monitored as previously
described, with some modifications.18 To determine IC50 of active
extracts, aliquots of 1.95 mL of a freshly prepared 0.2 mmol L12,2-
diphenyl-1-picrylhydrazyl (DPPH) colored solution in methanol
were mixed with 0.05 mL of the extract. The solutions were shaken
and incubated at 28 C for 2 h in the dark. A decrease in DPPH
absorbance induced by the sample was measured at 517 nm
against methanol as a blank.
A doseresponse curve for five prepared dilutions of each
extract was plotted to determine the IC50 values. Results were
expressed as standard equivalents using Trolox (TE), quercetin
(QE), ascorbic acid (VCE) and gallic acid (GAE) based on their
IC50 values. Moreover, the antiradical efficiency (AE =1/ IC50 ) was
In vitro cytotoxic assay
Methanolic extract and water fraction prepared from R. rugosa
petals were evaluated for their cytotoxic activities against cervical
(HeLa ECACC 93021013 human cervical epithelial cell line derived
from cervical carcinoma of a 31-year-old black female) and breast
cancer (T47D ECACC 85102201 human breast carcinoma cells) cell
lines. Normal human skin fibroblast cells in vitro (HSF primary cell
culture isolated from the skin a 25-year-old female, 5th passage)
were included in the cytotoxicity test as a control group.
Each cell line was inoculated at l04cells mL1density on a
microtiter plate (Nunc, Roskilde, Denmark) in RPMI 1640 medium
(Sigma, St. Louis, MO, USA) with 10% FBS (Sigma) and the extracts
were added at a concentration of 100 µgmL
1. The cultures were
incubated for 48 h under standard conditions (37 C, 5% CO2,
90% humidity). End-point determinations were performed with 5-
bromo-2-deoxyuridine (BrdU) labeling and detection kit III (Roche
Diagnosis, Mannheim, Germany) using an Elisa reader (Genesys
20, Thermo Spectronic, Madison, WI, USA).10
The growth percentage was evaluated spectrophotometrically
versus the untreated controls using the cell viability assay. The
results of each spectrophotometric measurement were expressed
as the percentage of dead cells compared to untreated samples.
All experiments were carried out in triplicate.
Antibacterial assay in vitro
The antibacterial potential of R. rugosa extracts has been
evaluated using the micro-broth dilution method, which enables
determinationof minimal inhibitory concentration(MIC) according
to the earlier described procedure.10 Eight reference strains,
including Gram-positive bacteria (Staphylococcus epidermidis
ATCC 12228, S. aureus ATCC 25923, Bacillus subtilis ATCC 6633,
Micrococcus luteus ATCC 10240) and Gram-negative bacteria
(Escherichia coli ATCC 25922, Klebsiella pneumoniae ATCC 13883,
Pseudomonas aeruginosa ATCC 9027, Proteusmirabilis ATCC 12453)
were used. The plant material was dissolved in dimethyl sulfide
(DMSO) and a series of the twofold dilutions, ranging from 0.02
to 2.5 mg mL1, was prepared in MuellerHinton broth (Biocorp,
Poland) in 96-well microtiter plates. The wells were inoculated
with the bacterial suspension (final inoculum size 106colony-
forming units, CFU mL1). Following 24 h incubation at 35 C, MIC
was defined as the lowest concentration of the extract at which
no visible growth was observed. DMSO was used as a negative
control, while as a positive control only plant material in broth
and broth with inocula was included. Gentamicin was used as a
reference compound. Minimal bactericidal concentrations (MBCs)
were determined by collecting 20 µL from each well with growth
inhibition, placing on to duplicate Mueller –Hinton agar plates and
incubating at 35 C for 24 h. MBC was defined as the lowest extract
concentration at which there was no bacterial growth.
Antifungal assay in vitro
Determination of antifungal activity of M (quantitatively dissolved
in DMSO) was carried out for two reference yeast strains derived
from the ATCC collection i.e. Candida albicans ATCC 10231 and
Candida parapsilosis ATCC 22019. MIC values were designated
using a serial twofold dilution method in 96-well microtiter plates
according to the method described above, with modifications of
the substrate, inoculum density and incubation conditions. Serial
dilutions of M were performed in MuellerHinton agar (Biocorp,
Poland) supplemented with 2% glucose. The wells were inoculated
with the yeast suspension (final inoculum size 1.5 ×105CFU mL1).
After 48 hincubation at 30 C,the MICswere assessed visuallyas the
lowest extract concentration showing complete growth inhibition.
Fluconazole was used as a reference compound. In order to
determine the minimal fungicidal concentration (MFC) of M, 20 µL
from each tube that showed growth inhibition was streaked onto
Sabouraud dextrose agar plates. After 48 h incubation at 30 C,
the MFC was assessed visually as the lowest drug concentration at
which there was no growth. Experiments were done in triplicate.
Representative data are presented.
Statistical analysis
The extracts were assayed in triplicate in each test. Data were
expressed as mean ±standard deviation of the independent
measurements. Statistical analysis was performed by use of
Statistica 6.0 and Excel. Significant differences were calculated
according to Duncan’s multiple range test. Differences at the 5%
level were considered statistically significant.
Content of the major groups of active ingredients in plant
Although R. rugosa petals are one of to the best studied parts of
the species, the data about their biologically active metabolites
are still scarce. Since petals are widely used for food, medicinal and
cosmetic products, it was decided to investigate their composition
and biological potential.
First, a preliminary analysis of the major groups of secondary
metabolites in the raw material was carried out. The content of
phenolic compounds (i.e. total phenolic content, total flavonoid,
phenolic acid and tannin content) was determined using
spectrophotometric methods. As a result, high phenolic content
in the raw material was found (107.44 mg g1dry plant material)
with tannins constituting a significant proportion (46.1 mg g1of
dry plant material). Considerable amounts of tannins were also
previously reported by other authors. 3,4,19 In our study, large
amounts of flavonoids and phenolic acids were also demonstrated
(3.56 and 8.88 mg g1dry weight, respectively).
Further spectrophotometric analyses showed the presence of
carotenoids in an amount of 1.39 µgg
1. Moreover, for the first
time the content of polysaccharides (4.63 mg g1dry weight; with
2.34 mg hexoses and 2.29 mg pentoses) was determined. Previous c
2013 Society of Chemical Industry J Sci Food Agric (2013)
Biological activity and chemical composition of rose petals
studies reported only the presence of polysaccharopeptide
complexes in the plant material.9,20
Composition of the methanolic extract and its fractions
Since petals were found to be a valuable source of polyphenols,
in the next step phenolic acids and flavonoid glycosides
were thoroughly analyzed. For this purpose, the LC-ESI-MS/MS
technique was used. Analyses were conducted on the basic
methanolic extract (M) and its three fractions of different
polarity (FE, OE and W), which were subsequently investigated
for biological activity. Seven phenolic acids and nine flavonoid
glycosides were qualitatively and quantitatively determined
(Tables 2 and 3).
In the methanolic extract four phenolic acids were determined,
including large amounts of gallic acid (9.55 mg to 1 g of dry extract)
followed by smaller amounts of protocatechuic, gentisic and p-
coumaric acids. Only the presence of gallic and protocatechuic
acid in petals has been earlier reported.9,21
Analysis of the fractions obtained by separation of the basic
extract provided additional information about the raw material.
The largest amounts of phenolic acids passed to the ether fraction
(FE), in which two other phenolic acids were determined, i.e. caffeic
and salicylic. Smaller quantities of phenolic acids were present in
the OE fraction; however only in OE a small amount of synapic
acid was found. The aqueous fraction (W) contained only relatively
small amounts of gallic and protocatechuic acids.
To our knowledge, the present study is the first one to report
caffeic, gentisic, salicylic, synapic and p-coumaric acids in R. rugosa
LC-ESI-MS/MS analysis of flavonoids revealed the pres-
ence of nine flavonoid glycosides, primarily quercetin (0.32
1.12 µgmg
1) and kaempferol derivatives (0.05– 0.14 µgmg
Results are shown in Table 3.
The M extract contained seven flavonoid glycosides, including
large amounts of rutin, isoquercitrin and avicularin (1.12, 1.10
and 0.54 µgmg
1, respectively). Tiliroside, astragalin, quercitrin
and kaempferol-3-O-rutinoside occurred in smaller amounts
(0.05 –0.32 µgmg
1of dry extract). After separation, the majority
of compounds moved to the OE fraction (2.20 µgmg
1of dry
extract). Moreover, two additional glycosides (hyperoside and
apigenin-7-O-glucoside) were found in the OE fraction. In the ether
fraction (FE), lower concentrations of flavonoids were observed
(1.60 µgmg
1of dry extract); however, considerable amounts of
tiliroside (not found in the OE fraction) were determined. In the
aqueous fraction no flavonoids were detected.
Avicularin, astragalin and hyperoside were previously found
in the plant material;21 however, this is the first time rutin,
isoquercitrin, tiliroside, quercitrin, kaempferol-3-O-rutinoside and
apigenin-7-O-glucoside have been reported in rugosa rose petals.
Antiradical activity
Considering earlier reports and our findings about high antiradical
activity of petals,2,8,22 it was decided to determine which fraction
wasresponsible for the methanolicextract activity. Foreach sample
(M, FE, OE and W) the concentration providing 50% scavenge of
the initial amount of DPPHradical under the conditions of the
assay used (IC50) was calculated from the graph of inhibition
percentage against extract concentration. Antiradical efficiency
(AE) was calculated (AE =1/IC50 ). The results were also expressed
as Trolox, quercetin and gallic acid equivalents (Table 4), which
enabled precise comparisons.
The study findings confirmed strong antiradical properties of
the petal methanolic extract (IC50 =0.80 mg mg1DPPH). The
analysis of activities of individual fractions obtained from the basic
methanolic extract separation showed varied antioxidant activity.
The acetate fraction (OE) had an extremely high antioxidant
potential. Its activity was 10 times higher than the basic extract
activity (IC50 =0.08 mg mg1DPPH), was found comparable to
the activity of gallic acid (GAE =1.03) and exceeded the activity
of Trolox, quercetin and ascorbic acid. Antioxidant potential of
the diethyl ether fraction (FE) was several times lower than OE
activity (IC50 =0.33 mg mg1DPPH), yet it was still very high
and comparable to Trolox potential (TE =0.95). Moreover, the
water extract showed the lowest, albeit significant activity (W;
IC50 =1.33 mg mg1DPPH).
The above results explicitly demonstrate that high petal
antioxidant activity depends largely on the compounds present
in the ethyl acetate fraction (most likely flavonoids). The ether
fraction (including large amounts of phenolic acids, e.g. gallic acid)
also plays a significant role, while compounds from the aqueous
fraction have the lowest impact on petal antioxidant effects.
Cytotoxic effect
Rose petals are known to contain compounds with potential
antiproliferative activity e.g. flavonoids, gallic and protocatechuic
acids, tannins,3,9,21,23 yet only a few studies noted this issue.
To our knowledge, information concerning cytotoxic activity of
R. rugosa petals can only be found in our previous report, which
demonstrated a significant viability decrease (9095%) in ovarian
(TOV-112D) and lung (A549) cancer line after exposure to galenic
preparations (tea and tincture) obtained from the plant material.
Additionally, preliminary analysis of tea anticancer activity against
cervical and breast cancer line was conducted showing a 50% and
25% viability decrease, respectively.10
Table 2. Content of phenolic acids in different extracts from rugosa rose petals. Mean values of three replicate assays with standard deviation
Phenolic acid (µgmg
1dry extract) M FE OE W
Caffeic acid nd 0.14±0.01 nd nd
Gentisic acid 0.03 ±0.00 0.09 ±0.00 0.02 ±0.00 nd
Protocatechuic acid 0.56 ±0.03 3.57 ±0.18 0.32 ±0.02 0.02 ±0.00
Gallic acid 9.55 ±0.48 16.46 ±0.82 7.96 ±0.38 0.27 ±0.02
Salicylic acid nd 0.13±0.01 nd nd
Synapic acid nd nd 0.01 ±0.00 nd
p-Coumaric acid 0.10 ±0.01 0.10 ±0.01 nd nd
SUM 10.24 ±0.51 21.39 ±1.07 8.31 ±0.41 0.29 ±0.02
Abbreviations: M, methanolic extract; FE, diethyl ether fraction; OE, ethyl acetate fraction; W, water fraction; nd, not detected.
J Sci Food Agric (2013) c
2013 Society of Chemical Industry R Nowak et al.
Table 3. Content of flavonoid glycosides in different extracts from rugosa rose petals. Mean values of three replicate assays with standard deviation
Flavonoid (µgmg
1dry extract) M FE OE W
Quercetin-3-L-arabinofuranoside (avicularin) 0.54 ±0.03 0.29 ±0.01 0.64 ±0.03 nd
Quercetin-3-O-rutinoside (rutin) 1.12 ±0.05 0.04 ±0.00 0.13 ±0.01 nd
Quercetin-3-O-galactoside (hyperoside) nd 0.04 ±0.00 0.05 ±0.00 nd
Quercetin-3-O-glucoside (isoquercitrin) 1.10 ±0.04 0.63 ±0.03 0.92 ±0.04 nd
Kaempferol-3-O-(6”-O-(E)-p-coumaroyl)-glucoside (tiliroside) 0.12 ±0.01 0.50 ±0.02 nd nd
Kaempferol-3-O-glucoside (astragalin) 0.14 ±0.01 0.04 ±0.00 0.05 ±0.00 nd
Quercetin-3-O-rhamnoside (quercitrin) 0.32 ±0.02 0.06 ±0.00 0.36 ±0.02 nd
Kaempferol-3-O-rutinoside 0.05 ±0.00 nd 0.02 ±0.00 nd
Apigenin-7-O-glucoside nd nd 0.03 ±0.00 nd
Sum 3.39 ±0.16 1.60 ±0.05 2.20 ±0.11 —
Abbreviations as in Table 2.
Therefore, our objective was to investigate antiproliferative
activity of the methanolic extract (M) against cervical (HeLa) and
breast (T47D) cancer cell lines. Moreover, the influence of M
on normal human skin fibroblasts was evaluated. The study was
conducted for three different concentrations of methanolic extract
(100, 75 and 50 µg of dry extract mL1). Results are presented in
Fig. 1. At a concentration of 100 µgmL
1, M showed an extremely
high antiproliferative effect on T47D and HeLa lines (90% and
100% of dead cells, respectively), yet the damaging effect on
normal fibroblast cell line was also significant (60% of dead cells).
At a concentration of 75 µgmL
1the methanolic extract effect
on T47D line was comparable to that obtained for a higher
concentration (90% of dead cells); however, a significant decrease
in the activity on the cervical line (50% of dead cells) and still
high fibroblast damage (50%) were observed. At the lowest
concentration tested (50 µgmL
1) M exhibited selective cytotoxic
effects, significantly inhibiting the proliferation of breast cancer
line (80% of dead cells) and slightly damaging HeLa cells (10%).
Furthermore, moderate toxic effects (25%) on skin fibroblasts were
To sum up, the extract cytotoxic effect on cervical (HeLa) and
normal human skin fibroblasts line decreased with dose reduction.
Such a relation was not observed in the case of T47D line, where
dose reduction had no significant impact on extract activity.
Additionally, antiproliferative activity of the water fraction (W)
on the same cancer cell lines was examined. The aim was to
check whether the water fraction (after extraction with diethyl
ether and ethyl acetate, i.e. after removing the majority of
phenolic acids and flavonoids) still exhibited cytotoxic activity.
Surprisingly, the results revealed a considerable viability decrease
(80% of dead cells) on both cancer lines after incubation with the
water fraction at a concentration of 100 µgmL
1. Moreover, no
toxic effect on the normal skin fibroblast cell line was observed.
This indicates a significant impact of compounds from the water
fraction on the petal cytotoxic potential. The activity of W may
be related to condensed tannins, polysaccharides, complexes of
polysaccharides with proteins or phenolic compounds, whose
presence was previously demonstrated in the material.3,9,19,20
Antimicrobial activity
In traditional Asian folk medicine, dried rugosa rose petals
have been mainly used due to their antibacterial properties;3,5,9
however, these properties have been poorly investigated so far.
Kamijo et al.19 examined the influence of pulverized Rosa rugosa
petals and its hydrolysable tannins (rugosin D and tellimagrandin
II) on the growth of intestinal and pathogenic bacteria, showing the
inhibition of growth of E. coli,S. aureus,B. cereus and Salmonella sp.
In addition, our previous analysis showed antibacterial potential
of tea and tincture prepared from R. rugosa petals.10 In the present
study, petal antimicrobial properties were examined in detail.
The methanolic extract and its two fractions (OE and W) were
screened for their antibacterial properties against eight reference
T 47D
100 µg mL
100 µg mL
75 µg mL-1 50 µg mL-1
T 47D
Figure 1. Results of cell proliferation assays obtained for methanolic extract (M; added at a concentration of 100, 75 and 50 µgmL
1) and water fraction
(W; added at concentration of 100 µgmL
1) after 48 h incubation. Data are expressed as percentage of dead cells. Error bars indicate standard deviation.
Abbreviations: HeLa, cervical cancer cell line; T47D, breast cancer cell line. c
2013 Society of Chemical Industry J Sci Food Agric (2013)
Biological activity and chemical composition of rose petals
Table 4. Antiradical activity of methanolic extract (M), diethyl ether (FE), ethyl acetate (OE) and water (W) fraction from Rosa rugosa petals
M 0.9952 0.80 1.24 10.30 2.34 5.25 3.04
FE 0.9967 0.33 3.06 4.20 0.95 2.14 1.24
OE 0.9974 0.08 12.44 1.03 0.23 0.53 0.30
W 0.9946 1.33 0.75 17.08 3.87 8.71 5.05
IC50 expressed as mg of dry extract per mg DPPH, mean values of three replications; AE, antiradical efficiency (1/IC50); GAE, gallic acid equivalent;
TE, Trolox equivalent; QE, quercetin equivalent; VCE, ascorbic acid equivalent. Equivalents were calculated by dividing extract IC50 by standard IC50.
For Trolox IC50 =0.344 mg mg1DPPH;forquercetinIC
50 =0.153 mg mg1DPPH; for gallic acid IC50 =0.078 mg mg1DPPH; for ascorbic acid
IC50 =0.264 mg mg1DPPH.
Table 5. Antibacterial and antifungal activity of different extracts obtained from Rosa rugosa petals
Reference microbial strain MIC MBC ratio MIC MBC ratio MIC MBC ratio
Gram-positive bacteria
S. epidermidis ATCC 12228 0.078 0.625 8 0.625 0.625 1 0.625 2.54
S. aureus ATCC 25923 0.313 2.5 8 0.313 0.625 2 1.25 2.52
B. subtilis ATCC 6633 0.313 1.25 4 0.313 0.625 2 1.25 2.52
M. luteus ATCC 10240 0.078 0.625 8 0.313 0.625 2 0.625 1.25 2
Gram-negative bacteria
E. coli ATCC 25922 0.625 1.25 2 0.313 0.625 2 0.625 1.25 2
K. pneumoniae ATCC 13883 0.625 1.25 2 0.313 2.580.625 2.54
P. aeruginosa ATCC 9027 1.25 1.25 1 0.313 0.625 2 0.625 1.25 2
P. mirabilis ATCC 12453 1.25 2.5 2 0.625 0.625 1 1.25 2.52
C. albicans ATCC 10231 0.156 1.258— — —
C. parapsilosis ATCC 22019 0.156 1.258— — —
Abbreviations: MIC, minimal inhibitory concentration (mg mL1); MBC, minimal bactericidal concentration (mg mL1); ATCC, American Type Culture
MICs of gentamicin ranged from 0.03 to 0.12 ×103mg mL1and from 0.25 to 1.0 ×10-3 mg mL1for Gram-positive and Gram-negative bacterial
strains, respectively. MIC for fluconazole 0.008 mg mL1. DMSO at the final concentration used had no influence on the growth of the tested strains;
—, not determined.
Gram-positive and Gram-negative bacteria. Additionally, the M
antifungal activity against two Candida species was studied. The
obtained data are presented in Table 5.
Our results showed that M, OE and W were active against all the
microorganisms used. In most cases, the basic methanolic extract
was found to be more active against Gram-positive bacteria than
its fractions, with MICs ranging from 0.078 to 0.313 mg mL1.Its
activity against M. luteus and S. epidermidis was particularly high
(MIC =0.078 mg mL1). Lower inhibitory effects were showed by
the OE fraction with MIC from 0.313 to 0.625 mg mL1, while the
activity of W was the lowest in the group (MIC from 0.625 to
1.25 mg mL-1).
This is likely to suggest that the activity against Gram-positive
bacteria is associated with synergistic activity of all different groups
on the compounds found in the methanolic extract.
In the case of Gram-negative bacteria, OE showed the best
antibacterial properties, whereas M and W exhibited lower and
very similar potential. This may indicate that the petal activity
against Gram-negative bacteria is affected by the acetate fraction
compounds, most likely flavonoids, which is in accordance with
the results obtained earlier.10
The activity of extracts was examined in detail, determining the
minimal bactericidal concentration (MBC) and comparing with MIC
values. Low MBC/MIC ratios (4) indicate the bactericidal activity.
According to the data (Table 5), almost all tested samples were
bactericidal, except for M towards S. epidermidis,S. aureus and M.
luteus and OE towards K. pneumoniae (MBC/MIC ratio =8).
The preliminary study of antifungal activity of M against two
Candida species showed considerable growth inhibition (MICs
0.156 mg mL1). MFC/MIC ratios (>8) indicate that methanolic
extracts possess fungistatic activity. This is the first report on petal
antifungal properties.
To sum up, such detailed studies on chemical composition and
biological activity of R. rugosa petals have been conducted
for the first time. Thus their chemical profile was determined,
indicating significant amounts of biologically active compounds,
polyphenols in particular. Five phenolic acids (caffeic, gentisic,
salicylic, synapic and p-coumaric) and six flavonoids (rutin,
isoquercitrin, tiliroside, quercitrin, kaempferol-3-O-rutinoside and
apigenin-7-O-glucoside), which have not been previously reported
in the plant material, were detected.
Based on biological activity assays, antioxidant, cytotoxic and
antimicrobial activities of petal methanolic extracts were revealed.
J Sci Food Agric (2013) c
2013 Society of Chemical Industry R Nowak et al.
Examinations of individual fractions obtained from M enabled
determination of which of them was responsible for a given type
of biological activity.
Our findings demonstrated that high petal antioxidant activity
depended largely on the compounds present in the ethyl acetate
fraction (probably mainly flavonoids).
Petals showed high antiproliferative effect on T47D and HeLa
cancer lines. The activity of the water fraction was of particular
interest, which had significant impact on petal antiproliferative
activity and did not contain substances damaging fibroblasts.
Additionally, R. rugosa petals turned out to possess notable
antibacterial and antifungal properties. However, it was not
possible to define explicitly which group of compounds was
responsible for this activity.
R. rugosa petals are a well-known and tasty addition to
the human daily diet. According to the results obtained, they
contain large amounts of biologically active ingredients. Owing
to their antiradical and cytotoxic properties, they deserve wider
dissemination and promotion as chemopreventive agents.
This work was financially supported by the Polish Ministry of
Science and Higher Education (grant no. N N405 617938). The
paper was developed using equipment purchased within the
Project ‘The equipment of innovative laboratories doing research
on new medicines used in the therapy of civilization and neoplastic
diseases’ within the Operational Program Development of Eastern
Poland 20072013, Priority Axis I Modern Economy, Operations
I.3 Innovation Promotion.
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Emergence of antibiotic-resistance pathogens has caused serious health issues and if the current trend is to continue, treatment of infection will become complicated and even unsuccessful due to new antimicrobial resistance (AMR). Therefore, there is a global drive to identify new methods to treat infection and develop better antibacterial materials and therapy. Although new and more potent antibiotics have aided the fight against microbes, they only offer a temporary solution because future bacteria strains may become resistant to these antibiotics and drugs. Recently, application of non-biological methods such electrical currents and photothermal/dynamic therapies to kill bacteria, reveals new approaches to design antimicrobial biomaterials, as complications stemming from drug-resistant bacteria can be obviated. Furthermore, recent research has focused on mimicking the surface patterns on plants and insects such as lotus leaves and dragonfly wings. Bio-inspired micro/nano patterns have been replicated on a variety of biomaterials to improve the bacterial resistance and other properties with good success. This is an exciting research area with immense practical and clinical potentials. In this review, recent advances in the application of chemical/biological approaches to combat bacterial infection and AMR are summarized and the related mechanisms are discussed. This article is protected by copyright. All rights reserved.
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Rosa rugosa Thumb., Rosa davurica Pall., and Rosa acicularis Lindl. contain a large number of target analytes which are bioactive compounds. High performance liquid chromatography (HPLC), in combination with the ion trap (tandem mass spectrometry), was used to identify target analytes in MeOH extracts of R. rugosa, R. davurica, and R. acicularis, originating from the Russian Far East, Trans-Baikal Region, and Western Siberia. The results of initial studies revealed the presence of 146 compounds, of which 115 were identified for the first time in the genus Rosa (family Rosaceae). The newly identified metabolites belonged to 18 classes, including 14 phenolic acids and their conjugates, 18 flavones, 7 flavonols, 2 flavan-3-ols, 2 flavanones, 3 stilbenes, 2 coumarins, 2 lignans, 9 anthocyanins, 3 tannins, 8 terpenoids, 3 sceletium alkaloids, 4 fatty acids, 2 sterols, 2 carotenoids, 3 oxylipins, 3 amino acids, 5 carboxylic acids, etc. The proven richness of the bioactive components of targeted extracts of R. rugosa, R. davurica, and R. acicularis invites extensive biotechnological and pharmaceutical research, which can make a significant contribution both in the field of functional and enriched nutrition, and in the field of cosmetology and pharmacy.
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The demand for healthy, viable, and safer products are an ever‐increasing trend in the global market. The usage of flowers as food, traditional cuisine, or alternative medicine is very common in various regions of the world because of their significant nutritional profile and therapeutic potential as demonstrated by various studies. In this context, the marketing segment of these edible flowers in the form of food is increasing owing to their beneficial impact on human health. The presence of the various bioactive compounds in edible flowers has attracted major industries like food and pharmaceutical companies for the development of functional foods and medicine. There is a dire need to explore the concealed aspects of edible flowers on a commercial level to get economic and health benefits. The present review deals with the availability of various edible flowers like heartsease, common daisy, marigold, evening primrose and boraginaceae etc and elucidate their phytochemical screening, nutritional, bioactive, medicinal, chemical features along with consumptional behavior.
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The study was designed to determine the total phenolic, flavonoid, o-dihydroxyphenol, tannin, and carotenoid content as well as the antiradical, antitumor and antimicrobial properties of two types of galenic preparations from Rosa rugosa Thunb. Such extracts obtained from various plant parts have not been studied to date. Our findings have revealed high antiradical activity of the examined galenic preparations, with root, leaf and flower extracts (IC50 ranging from 0.27 to 0.19 mg of dry extract per mg DPPH·) showing the greatest potential. MIC and MBC values against 8 reference bacterial strains (i.e. Staphylococcus epidermidis, Staphylococcus aureus, Bacillus subtilis, Micrococcus luteus, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Proteus mirabilis) were determined. Generally, tinctures were found to be more active than teas with MIC ranging from 0.08 to 2.5 mg mL−1 and 0.31 to 1.25 mg mL−1, respectively. Anticancer activities against ovarian (TOV-112D), cervical (HeLa), breast (T47D) and lung cancer (A549) cell lines were evaluated using the BrdU test. The data obtained demonstrate considerable impact of polyphenols on the anticancer activity of extracts (ethanolic, in particular).
The effects of a methanol extract of Rosa rugosa root and its triterpenoid glycoside, rosamultin, on hepatic lipid peroxidation and drug-metabolizing enzymes were investigated in rats treated with bromobenzene. The methanol extract of R. rugoso root reduced the activities of aminopyrine N-demethylase and aniline hydroxylase, which had been increased by bromobenzene, but rosamultin did not affect the activities of the two enzymes. Both the methanol extract and rosamultin restored the activity of epoxide hydrolase, which had also been decreased by bromobenzene. Hepatic glutathione concentrations were lowered and hepatic lipid peroxides were increased in rats intoxicated with bromobenzene. The hepatic lipid peroxidation induced by bromobenzene was prevented with the methanol extract and rosamultin. However, the decrease in glutathione was not altered by the methanol extract of R. rugosa. These results suggest that the extract of R. rugosa and its compound, rosamultin, may protect against bromobenzene-induced hepatotoxicity through, at least in part, enhanced activity of epoxide hydrolase. Antioxidant properties may contribute to the protection of R. rugosa against bromobenzene-induced hepatotoxicity.
A novel, easy, and cheap technique for preliminary quantitative evaluation of antiradical activity, based on HPTLC, has been proposed. This method combines chromatographic separation of polar compounds, present in plant extracts, with data analysis by means of image processing software. Bleaching of the purple DPPH• color, caused by substances with antiradical activity, was observed and recorded using a photo camera. ImageJ, a free and open source image processing program was used for quantitative measurements. For evaluation of assay efficiency, the antiradical activity of rose flower extracts (from Rosa rugosa Thunb.) was expressed as Standard Activity Coefficients (SACs), which are relative measures of the activity to the four well known antioxidants; i.e., quercetin, gallic acid, protocatechuic acid, and Trolox. The method uses small amounts of free radical and is easily applicable – only a digital camera with freely available open source software is required.
The scavenging activities of the crude aqueous extracts from 69 kinds of fresh flowers in southern China on DPPH, superoxide and hydroxyl free radicals, and their polyphenolic contents, were investigated and evaluated in the paper. The results showed that the extracts of red rose flowers had obviously stronger antioxidant activity when compared to other flowers. Polyphenolic content had significant relationships with the 1,1-diphenyl-2-picrylhydrazyl (DPPH) (r = 0.983, P < 0.01) and superoxide free radical scavenging activity (FRSA) (r = 0.588, P < 0.01), but no significant relationship with hydroxyl FRSA (r = 0.184, P > 0.05) in tested open flowers. However, in tested flower buds, polyphenolic content was correlated well with the DPPH FRSA (r = 0.993, P < 0.01), superoxide FRSA (r = 0.738, P < 0.01), and hydroxyl FRSA (r = 0.737, P < 0.0 1), respectively. Rose was found to be a promising resource of antioxidant polyphenolics that might contribute to the development of functional food. (C) 2007 Published by Elsevier Ltd on behalf of Swiss Society of Food Science and Technology.
Two kinds of rose (Rosa damascena Miller) oil were used: decanted oil distilled from the fresh petals (DOFP) and blended oil distilled from the paled (fermented) petals (BOPP). Eight samples from Gülbirlik cooperative factories (three DOFP, four BOPP) and a private factor (one BOPP) were of 1990 produce, and seven samples from Gülbirlik (two DOFP, four BOPP) and a private factory (one BOPP) were of 1991 produce. In all, 15 samples were analysed for density, refractive index and optical rotation, and their chemical composition by GC and GC-MS. The samples showed some differences in the optical rotation. DOFP had high density and refractive index values. In general, 68 components were identified, representing c 88–98% of the oils: 27 hydrocarbons, 19 alcohols, eight aldehydes, six oxides and ethers, five esters, two ketones and one phenol. Citronellol (24.47–42.97%), nonadecane (6.44–18.95%), geraniol (2.11–18.04%), ethanol (0.00–13.43%), heneicosane (2.28–8.90%), nerol (0.75–7.57%) and 1-nonadecene (1.80–5.40%) were the major components as overall ranges of the samples. Percentages of certain components differed between the factories, years and oil types (DOFP and BOPP). The results indicated that an extensive and detailed research from harvesting to end-product storage was needed for the establishment of the material and technology requirements to obtain a Turkish rose oil with standard composition and high quality.