Journal of Ethnopharmacology 120 (2008) 378381

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Journal of Ethnopharmacology
journal homepage: www.elsevier.com/locate/jethpharm

Evaluation of anti-inammatory and antinociceptive activities of some

Onosma L. species growing in Turkey
Alev Tosun a , Esra Kpeli Akkol b , zlem Bahadr a , Erdem Yesilada c,
a
Department of Pharmacognosy, Faculty of Pharmacy, Ankara University, Tandogan 06100, Ankara, Turkey
b
Department of Pharmacognosy, Faculty of Pharmacy, Gazi University, Etiler 06330, Ankara, Turkey
c
Department of Pharmacognosy, Faculty of Pharmacy, Yeditepe University, Kayisdagi 34755, Istanbul, Turkey

a r t i c l e i n f o a b s t r a c t

Article history: Ethnopharmacological relevance: Roots of Onosma species are used for the treatment of various disorders
Received 5 May 2008 such as bronchitis, tonsillitis, hemorrhoids as well as alleviating pains in folk medicine in Turkey.
Received in revised form 4 September 2008 Aim of the study: The chloroform and ethanol (70%) extracts obtained from the roots of Onosma species
Accepted 5 September 2008
(Boraginaceae) growing in Turkey, Onosma aucheranum DC., Onosma isauricum Boiss. and Heldr. (endemic),
Available online 18 September 2008
Onosma sericeum Willd., Onosma tauricum Pallas ex Willd. var. brevifolium DC. (endemic) and Onosma
tauricum Pallas ex Willd. var. tauricum (Syn: Onosma velenovskyi Davidov) were evaluated for in vivo
Keywords:
anti-inammatory and antinociceptive activities.
Anti-inammatory activity
Antinociceptive activity
Materials and methods: For the preliminary screening, carrageenan-induced hind paw edema for the anti-
Onosma species inammatory activity and, p-benzoquinone-induced abdominal constriction test for the antinociceptive
Boraginaceae activity were used in mice.
Results: The chloroform extracts from Onosma aucheranum and Onosma isauricum and ethanolic extracts
from Onosma isauricum and Onosma sericeum demonstrated 28.0%, 34.3%, 24.6% and, 27.5% inhibition
in p-benzoquinone-induced abdominal constriction test. The chloroform and ethanol (70%) extracts of
Onosma isauricum and ethanol (70%) extract of Onosma sericeum also exhibited marked inhibition, rang-
ing between 12.327.3%, 10.525.3%, 8.222.6%, respectively, in carrageenan-induced hind paw edema
model at 100 mg/kg dose without gastric damage and the activity was quite comparable to indomethacin
(32.038.4% inhibition) as a reference sample. Neither death nor gastric bleeding was observed for any of
the plant extracts during the acute toxicity evaluation.
Conclusion: The experimental data demonstrated that Onosma aucheranum, Onosma isauricum and Onosma
sericeum displayed remarkable anti-inammatory and antinociceptive activities.
2008 Published by Elsevier Ireland Ltd.

1. Introduction
Onosma species (Boraginaceae) widely grow in Turkey. Eighty-
ve species including 44 endemic species has been represented so
far (Riedl, 1978). The genus has several utilizations in the tradi-
tional medicines worldwide. It is noted that the roots of Onosma
species are used for the treatment of various disorders such as
bronchitis, tonsillitis, hemorrhoids as well as alleviating pains in
folk medicine in Turkey (zgen and Coskun, 2001; zgen et al.,
2006; Sezik et al., 1997). The root has the important usage regard-
ing the recovery of wounds and burns. The graded roots or its
barks from Alkanna or Onosma species are prepared as an ointment
Corresponding author. Fax: +90 216 5780068.
E-mail address: yesilada@yeditepe.edu.tr (E. Yesilada).
and kept as a home remedy to treat wounds, burns and hemor-
rhoids, externally (zgen et al., 2006; Sezik et al., 1997). In addition
Onosma hispida Wall. can be used for laxative and anthelmintic pur-
poses and is also treat eye diseases, blood disorders, in bronchitis,
abdominal pains, as well as against thirst, itching, fever, wounds,
piles, dysuria and urinary calculi. The plant owers are prescribed
as a cardiac stimulant on heart as well as against rheumatism
(Ahmad et al., 2005b).
As evidenced from the above assertions, Onosma species is
used in traditional medicines worldwide. In the present study,
in vivo anti-inammatory and antinociceptive activities of the
root extracts of several Onosma species (Onosma aucheranum DC.,
Onosma isauricum Boiss. and Heldr. (endemic), Onosma sericeum
Willd., Onosma tauricum Pallas ex Willd. var. brevifolium DC.
(endemic) and Onosma tauricum Pallas ex Willd. var. tauricum (Syn:
Onosma velenovskyi Davidov)) in Turkey were investigated for the
traditional utilizations.

0378-8741/$ see front matter 2008 Published by Elsevier Ireland Ltd.

doi:10.1016/j.jep.2008.09.007
 A. Tosun et al. / Journal of Ethnopharmacology 120 (2008) 378381
2. Experimental
2.1. Plant materials
Plant materials were collected from different places in Turkey.
The specimens of voucher were authenticated by Prof. H. Duman
of Department of Biology, Faculty of Sciences & Letter, Gazi Univer-
sity, and were deposited at the Herbarium of Faculty of Pharmacy of
Ankara University (AEF). Collection sites and herbarium numbers
are as follows: Onosma aucheranum DC. (23684): Ankara-Cubuk-
Karagl, 19/05/2005. Onosma isauricum Boiss. and Heldr. (23686)
(endemic, Ekim et al., 2000): Ankara-Cubuk-Karagl, 19/05/2005.
Onosma sericeum Willd. (23683): Konya-Ermenek, 25/06/2005.
Onosma tauricum Pallas ex Willd. var. brevifolium DC. (23687)
(endemic, Ekim et al., 2000): Ankara-Camldere, 22/05/2005.
Onosma tauricum Pallas ex Willd. var. tauricum (23688): Ankara-
Konutkent, 05/05/2006 (Table 1).
2.2. Preparation of plant extracts
The plant roots were dried under shade and powdered by using
a laboratory scale mill. The extracts were prepared as following
procedures.
The chloroform (CHCl3 ) extract: The plant material (10 g) was
extracted with chloroform (200 ml) under reux for 3 h. The
extract was evaporated in vacuum to give crude CHCl3 extract.
The ethanol (EtOH) extract: The plant material (10 g) was
extracted with ethanol (70%) under reux (200 ml) for 3 h. The
extract was evaporated in vacuum to give crude EtOH extract.
2.3. Pharmacological procedures
2.3.1. Animals
Male Swiss albino mice (2025 g) were purchased from the ani-
mal care laboratories (Rek Saydam Central Institute of Health,
Ankara, Turkey). The animals were maintained on standard pel-
let diet and water ad libitum. The food was withdrawn on the day
before the experiment, but allowed free access of water. A number
of animals were at least six from each group. Animals were pro-
cessed according to the suggested ethical international guidelines
for the care of laboratory animals of Gazi University Animal Ethic
Committee.
2.3.2. Preparation of test samples for bioassay
Plant extracts were given in 100 mg/kg dose after suspending
in 0.5% sodium carboxymethyl cellulose (CMC) suspension with
distilled water. The control group animals received the same exper-
imental handling as those of the test groups except that the drug
treatment was replaced with appropriate volumes of the dosing
vehicle. Either indomethacin (10 mg/kg) or acetyl salicylic acid
(ASA, 100 mg/kg) in 0.5% CMC was used as reference.
2.3.3. Antinociceptive activity
According to the method of p-benzoquinone-induced abdom-
inal constriction test (Okun et al., 1963) 60 min after given test
Table 1
The Onosma species studied, collection sites, herbarium numbers (AEF) and percentage yields of CHCl3 and EtOH extracts.
Species
Onosma aucheranum DC.
Onosma isauricum Boiss. and Heldr. (endemic)
Onosma sericeum Willd.
Onosma tauricum Pallas ex Willd. var. brevifolium DC. (endemic)
Onosma tauricum Pallas ex Willd.var. tauricum
379
samples, the mice were injected with 0.1 ml/10 g body weight of
2.5% (w/v) p-benzoquinone (i.p., PBQ; Merck) solution in distilled
H2 O. Control animals received an appropriate volume of dosing
vehicle. The mice were then kept individually in cages and the
total number of abdominal contractions (writhing movements) was
counted for the next 15 min as starting on the 5th-min. The data
represent average of the total number of writhes observed. The
antinociceptive activity was expressed as percentage of writhing
controls. ASA (100 mg/kg) was used as the reference.
2.3.4. Anti-inammatory activity
Carrageenan-induced hind paw edema model was used for
the assessment of anti-inammatory effect (Yesilada and Kpeli,
2002). Sixty minutes after test sample suspension of carrageenan
(0.5 mg/25 l in 154 nM NaCl) was injected into each sub-plantar
tissue of the right hind paw. As the control, 25 l saline solu-
tions were injected into that of the left hind paw. Paw edema was
measured in every 90 min during 6 h after induction of inamma-
tion. The difference in footpad thickness was measured by a gauge
calipers (Ozaki co., Tokyo, Japan). Indomethacin (10 mg/kg) was
used as the reference drug. Mean values of treated groups were
compared with mean values of a control group and analyzed using
statistical methods.
2.3.5. Acute toxicity
Animals in the carrageenan-induced paw edema experiment
were observed during 48 h as well as morbidity or mortality was
recorded (Yesilada and Kpeli, 2002).
2.3.6. Gastriculcerogenic effect
After the measurements of antinociceptive activity, mice were
killed with ether anesthesia and stomachs were removed. The
abdomen was opened by the greater curvature and dissected with
microscope in order to observe lesions or bleedings (Yesilada and
Kpeli, 2002).
2.3.7. Statistical analysis of data
Data were expressed as mean (S.E.M.). Statistical differences
were evaluated by ANOVA and StudentsNewmanKeuls post hoc
tests. p < 0.05 was considered to be signicant (*p < 0.05; **p < 0.01;
***p < 0.001).
3. Results and discussion
The chloroform and ethanolic (70%) extracts of plant roots were
evaluated for anti-inammatory activity and antinociceptive activ-
ity.
As illustrated in Table 2, the chloroform extracts of Onosma
aucheranum and Onosma isauricum and ethanolic extracts of
Onosma isauricum and Onosma sericeum displayed signicant
antinociceptive activity; 28.0%, 34.3%, 24.6%, and 27.5% inhibition,
respectively, against p-benzoquinone-induced abdominal contrac-
tions, without inducing any apparent gastric damage. While ASA
as reference drug showed and inhibition by 52.6% inhibition at the
same dose, ve mice suffered from severe gastric damage.
Collection sites and dates AEF No: CHCl3 extract (w/w, %) EtOH extract (w/w, %)
Ankara-Cubuk-Karagl 19/05/05 23684 0.1456 1.3778
Ankara-Cubuk-Karagl 19/05/05 23686 0.1028 1.0391
Konya-Ermenek 25/06/05 23683 0.0781 0.9575
Ankara-Camldere 22/05/05 23687 0.0805 0.8540
Ankara-Konutkent 05/05/2006 23688 0.1198 1.1107
380 A. Tosun et al. / Journal of Ethnopharmacology 120 (2008) 378381

Table 2
Effect of the extracts against p-benzoquinone-induced writhings in mice.

Material Extract type Dose (mg/kg) Number of writhings SEM Inhibitory ratio (%) Ratio of ulceration

Control 51.3 4.4 0/6

Onosma aucheranum CHCl3 100 36.9 2.9 28.0* 0/6

200 35.2 3.4 31.4** 0/6
EtOH 100 48.5 4.2 5.5 0/6
200 49.2 2.9 4.1 1/6

Onosma isauricum CHCl3 100 33.7 3.2 34.3** 0/6

200 35.9 2.6 30.0** 0/6
EtOH 100 38.7 2.1 24.6* 0/6
200 36.1 4.2 29.6* 0/6

Onosma sericeum CHCl3 100 45.7 2.4 10.9 0/6

200 49.9 5.1 2.7 2/6
EtOH 100 37.2 2.7 27.5* 0/6
200 35.4 1.9 30.9** 0/6

Onosma tauricum var. brevifolia CHCl3 100 48.1 2.3 6.2 0/6
200 52.4 3.5 0/6
EtOH 100 46.3 2.2 9.7 0/6
200 48.6 5.6 5.3 0/6

Onosma tauricum var. tauricum CHCl3 100 43.3 3.1 15.6 0/6
200 45.8 3.9 10.7 0/6
EtOH 100 41.1 2.3 19.9 0/6
200 44.6 3.4 13.1 0/6

ASA 100 24.3 2.2 52.6*** 5/6

200 20.9 1.7 59.3*** 6/6

CHCl3 : chloroform extract; EtOH: ethanol extract (70%); S.E.M.: standard error mean.
*
p < 0.05.
**
p < 0.01.
***
p < 0.001.

The chloroform and ethanolic extracts of Onosma isauricum at 100 and 200 mg/kg doses, when compared to indomethacin ref-
and ethanolic extract of Onosma sericeum also exhibited notable erence compound (32.038.4% inhibition) (Table 3). All extracts
inhibition, ranging between 12.327.3%, 10.525.3% and 8.222.6%, were safe in the administered doses based on the assessment of
respectively, against carrageenan-induced hind paw edema model acute toxicity.

Table 3
Effects of the extracts against carrageenan-induced paw edema in mice.

Material Extract type Dose mg/kg Swelling thickness (102 mm) S.E.M. (% inhibition)

90 min 180 min 270 min 360 min

Control 49.7 3.3 55.3 3.9 59.7 4.8 65.9 4.1

Onosma aucheranum CHCl3 100 47.8 4.2 (3.8) 49.2 3.6 (11.0) 51.2 3.9 (14.2) 53.3 4.0 (19.1)
200 45.2 3.4 (9.1) 47.9 3.1 (13.4) 54.1 3.7 (9.4) 54.0 2.9 (18.1)
EtOH 100 50.2 2.6 57.8 2.9 61.3 3.5 66.9 3.0
200 49.8 4.5 56.5 3.4 60.8 3.9 69.2 3.1

Onosma isauricum CHCl3 100 43.6 2.6 (12.3) 45.2 2.3 (18.3) 44.7 2.1 (25.1)* 47.9 2.4 (27.3)**
200 41.4 3.0 (16.7) 43.7 3.4 (20.9) 41.5 2.9 (30.5)** 46.4 3.3 (29.6)**
EtOH 100 44.5 3.0 (10.5) 45.9 3.2 (16.9) 44.9 3.7 (24.8)* 49.2 3.0 (25.3)*
200 45.6 2.8 (8.2) 47.9 3.4 (13.4) 46.2 3.1 (22.6) 48.4 3.5 (26.6)*

Onosma sericeum CHCl3 100 51.3 4.2 56.9 4.9 61.2 3.8 68.7 3.9
200 54.2 5.1 59.8 3.7 62.9 4.6 70.5 3.8
EtOH 100 45.6 3.2 (8.2) 46.7 3.8 (15.6) 45.9 3.9 (23.1) 51.0 2.6 (22.6)*
200 44.3 2.6 (10.9) 45.1 2.1 (18.4) 44.6 3.4 (25.3)* 50.2 2.9 (23.8)*

Onosma tauricum var. brevifolia CHCl3 100 51.2 2.8 58.9 3.2 61.2 3.3 67.9 3.9
200 54.1 3.7 57.4 3.9 61.9 3.1 68.2 3.5
EtOH 100 58.3 1.9 61.8 2.9 65.6 3.4 71.8 3.0
200 48.6 2.3 (2.2) 53.8 2.9 (2.7) 55.6 3.6 (6.9) 60.4 3.4 (8.3)

Onosma tauricum var. tauricum CHCl3 100 47.8 4.1 (3.8) 54.3 3.9 (1.8) 58.9 3.8 (1.3) 63.3 4.0 (3.9)
200 60.2 3.0 64.8 3.7 69.1 4.3 75.6 3.4
EtOH 100 47.3 2.3 (4.8) 51.2 2.7 (7.4) 55.6 2.9 (6.9) 59.6 2.8 (9.6)
200 50.1 3.8 53.5 3.2 (3.3) 52.9 3.6 (11.4) 60.5 3.4 (8.2)

Indomethacin 10 31.5 2.3 (36.6)** 37.6 2.7 (32.0)** 38.2 1.8 (36.0)*** 40.6 2.6 (38.4)***

CHCl3 : chloroform extract; EtOH: ethanol extract (70%); S.E.M.: standard error mean.
*
p < 0.05.
**
p < 0.01.
***
p < 0.001.
 A. Tosun et al. / Journal of Ethnopharmacology 120 (2008) 378381
A number of studies have been conducted on the biologi-
cal effects and chemical compositions of several Onosma species.
Acetyl cholinesterase and butyryl cholinesterase enzyme inhibitory
activities of Onosma hispida Wall., has been demonstrated by
Ahmad et al. (2003) while the activity was attributed for its a-
vanone and benzoic acid derivatives. Onosmins as avonoid type
compounds inhibited lipoxygenase enzyme activity (Ahmad et
al., 2005b). Ethanolic extract of Onosma hispidum Wall., and its
fractions as well as ferulic and vanillic acids isolated from this
plant had signicant activity on several bacterial strains (Naz
et al., 2006). A signicant reduction in the number of tumors
was determined by the methanolic extract of Onosma echioides
L. on experimental skin cancer in mice (Sharma et al., 2004).
In a bioassay study, shikonin derivatives were isolated from the
cyclohexane extract of Onosma leptantha Held. root which is
the most active fraction for carrageenan-induced rat paw edema
test. The extracts and isolated components also exerted cyto-
toxic activity on both L1210 murine lymphoblastic leukemia cell
lines and human brosarcoma HT-1080 cells (Kundakovic et al.,
2006). The n-hexanedichloromethane extract from the roots
of Onosma argentatum Hub.-Mor. and four shikonin derivatives
(deoxyshikonin, acetyl shikonin, 3-hydroxy-isovaleryl shikonin and
5,8-O-dimethyl acetyl shikonin) were investigated for their abil-
ity to stimulate the growth of human amnion broblast (zgen
et al., 2006). The extract and 5,8-O-dimethyl acetyl shikonin were
active in 0.1 and 0.055 g/mL concentrations while all samples
were cytotoxic in 10 g/mL concentrations. Further this plant was
studied for its antioxidant and antimicrobial activities (zgen et al.,
2003). Its extract exhibited a potent antioxidant activity and effec-
tiveness on Staphylococcus aureus, Bacillus subtilis and Escherichia
coli. A signicant antiulcer activity of Onosma armeniacum K. root
extract against ethanol-induced lesions has been reported (Cadirci
et al., 2007). This effect attributed partially to its the antioxidant
effect of the extract. In addition, several pyrrolizidine alkaloids from
Onosma leptantha had hepatotoxic activity (Kretsi et al., 2003).
To date, several chemical constituents including naph-
thaquinones have been isolated from Onosma argentatum Hub.-
Mor. (zgen et al., 2004), pyrrolizidine alkaloids from Onosma
arenaria Waldst. and Kit. (El-Shazly et al., 2003), Onosma lep-
tantha Heldr. (Kundakovic et al., 2006) and Onosma stellatum
W.K. (Mroczek et al., 2004), lipids from Onosma heterophylla
Griseb (Mellidis and Papageorgiou, 1987; Papageorgiou and
Assimopoulou, 2003), ketones and triterpenoids from Onosma lim-
itaneum I.M. Johnston (Ahmad et al., 2005a). Recently, the essential
oil composition of Onosma microcarpum DC., was also investigated
by GC and GC/MS (Morteza-Semnani et al., 2006).
In the root barks of several Onosma species accumulation of
naphthaquinones of alkannin and shikonin derivatives have been
reported. These compounds possess signicant anti-inammatory
activity by the assessment of inhibitory effects on the increased
capillary permeability and thermal edema (Tanaka et al., 1986)
using carrageenan-induced paw edema tests in rats (Kundakovic
et al., 2006).
These data encouraged us to examine anti-inammatory and
antinociceptive activities of Onosma species and, Onosma aucher-
anum DC., Onosma isauricum Boiss. and Heldr., Onosma sericeum
Willd., Onosma tauricum Pallas ex Willd. var. brevifolium DC. and
Onosma tauricum Pallas ex Willd. var. tauricum (Syn: Onosma velen-
ovskyi Davidov) were investigated for their in vivo effects. The
381
traditional usages were supported by this study. Probably we sug-
gest that their activity may be the reason for their naphthaquinone
contents located in root barks of these plants. Future studies need
to isolate active compounds which are responsible for the activity
of these species.
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