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Investigation of common butterbur
(Petasytes hybridus L., Asteraceae) non-polar extract
Dušanka Runjaić-Antić1, Vanja Tadić2, Ivana Arsic3, Zorica Vujić3
2Alternativa medica, Manufacturer of phytopreparations, Loznica, Serbia;
4Institute for Medicinal Plant Research „Dr Josif Pančić“, Belgrade, Serbia; 1Faculty of Medicine,Department of Pharmacy, University of Niš, Serbia;
3Faculty of Pharmacy, University of Belgrade, Serbia;
Common butterbur (Petasytes hybridus L., Asteraceae) has
numerous applications and expanding clinical interest in
the treatment of migraines, allergic rhinitis, and asthma
(spasmolytic action). The combined anti-spasmodic, anti-
inflammatory and calcium-channel blocking effects of P.
hybridus proved a rationale base for its use in the
prophylaxis of migraine. The rhizomes (Petasitidis
rhizoma), and leaves (Petasitidis folium) are used for
medicinal purposes. Butterbur extracts contain
sesquiterpene esters, polifenolic acids, flavonoids, lignans,
essential oils and in the minor concentration pyrrolizidine
alkaloids. The literature data revealed that the main
constituents of butterbur are sesquiterpene esters of
petasin and isopetasin.
The plant material was collected on the mountain Jagodnja (Serbia).
The extraction procedure was performed with methylene chloride, and obtained
extract was analyzed for total polifenolic compounds (mg GA/g dry mass),
flavonoids (%), tannins (%) and petasine (%) content. The total phenolics content-TF
was determined by the Folin-Ciocalteu method. The percentage content of flavonoids
CF was calculated using the method described in DAB 10. The percentage content of
tannins-TC ware investigated using the method described in the Ph. Eur. 6.0. (Table
1). The total sesquiterpene content were determined according to Ph. Eor. 6.0, taking
S-petasin as reference standard. HPLC fingerprint of the extract (Figure 2 and 3) was
achieved by HPLC (Agilent Technologies 1200). Detection was performed using
Diode Array Detector (DAD), and the chromatograms were recorded at λ = 260 and
325 nm (for sesquiterpene lactones – petasines, and poliphenolic compounds,
respectively). HPLC separation of components was achieved using a LiChrospher
100 RP 18e (5 μm), 250 × 4 mm i.d. column, with a flow rate of 1 mL/min and
mobile phase, A [500 mL of H2O plus 9.8 mL of 85% H3PO4 (w/w)], B (MeCN),
elution, combination of gradient mode: 90-75% A, 0-25 min; isocratic 75% A, 25-30
min; 75-55% A, 30-46 min). The sample was prepared dissolving 118.6mg of the
extract in 10 mL of MeOH, filtered through 0.2 μm PTFE filters prior to HPLC
analysis. The injected volume was 4 μL. Standard solution of S-petasin was prepared
at a final concentration of 0.01 mg/mL.
Figure. 2. Comparative HPLC spectra of
S-petasin (a), MeOH (b) and CH2Cl2 (c)
common butterbur extracts (from 55-
65min). UV spectra of S-petasin (d), and
the compounds with Rt close to Rt of S-
petasin (e and f, from MeOH and CH2Cl2
extracts respectively)
Conclusion Further investigation should proceed in aim to determine the main
sesquiterpene lacones present in the species from this geografic region.
Besides, the profile of phenolic compounds would be a challenge to
determined, as well.
For more information please do not hesitate to contact us at:
E-mail: [email protected]
Phone: +381 11 7120 982
Fax.: + 381 11 3120 381
The aim of our investigation was to determine the chemical
profile of Petasitides rhizoma non-polar extract regarding
the content of total polifenolic compounds, including
flavonoids and tannins and total petasine content.
Investigated extract subjected to HPLC analysis revealed the absence of S-
petasine, as well as in the MeOH extracts, made to check the obtained
results (Figure 2).
min56 58 60 62 64
mAU
0
200
400
DAD1 C, Sig=260,8 Ref=600,100 (VMT\VT_01042013 2013-04-01 14-14-25\PETASH000100.D)
57.93
5 58.38
3
59.45
7
59.84
8
min56 58 60 62 64
mAU
0
25
50
DAD1 C, Sig=260,8 Ref=600,100 (VMT\VT_01042013 2013-04-01 14-14-25\PETASH000106.D)
55.27
2
56.53
0
57.06
7
58.18
2
58.89
2 59.61
4
60.46
1
61.02
0
62.36
6
62.81
9
64.44
8
min56 58 60 62 64
mAU
0
50
100
DAD1 C, Sig=260,8 Ref=600,100 (VMT\VT_01042013 2013-04-01 14-14-25\PETASH000112.D)
54.50
1
55.15
5
56.40
8
57.13
4
57.73
2
58.23
4
58.98
0 59
.207
59.59
9
60.05
8
60.53
6
61.10
9
62.00
4
62.48
2
63.02
1
63.35
7
64.53
0
nm250 300 350 400 450 500 550 600
mAU
0
50
100
150
200
DAD1, 57.886 (192 mAU, - ) of PETASH000100.D DAD1, 57.933 (235 mAU, - ) of PETASH000100.D
a
b
c
d
e
f
Introduction
Aim
Material and method
Results
Petsytidis rhizoma (Petasytes hybridus L., Asteraceae)
Total phenols (mg GA/g ekstrakta) 29.00
Flavonoids content (%) 0.10
Tannins content (%) 0.89
Sesquiterpene lactones content 2.10
Table 1. Total phenolic, tannins, flavonoid and
sesquiterpene lactones content in non-polar extract of
common butterbur root (Petasites hybridus, L.,
Asteraceae). The results were expressed by dry
weight.
Literature:
A. Bodensieck, O. Kunert, E. Haslinger, R. Bauer. Helv Chim Acta (2007), 90, 183-194; 2. R.
Chizzola, T. Langer, C. Franz. Planta Med. 2006, 72, 1254-1256
Blois, M.S., 1958. Antioxidant determinations by the use of a stable free radical. Nature. 181, 1199-
1200.
European Pharmacopoeia. 6th edition. 2004. Strasbourg: Council of Europe.
Weißdornblätter mit Blüten. In: Deutsches Arzneibuch, Band 2, Monographien R-Z; Amlitche
Ausgabe Deutcher Apotheker Verlag. Stuttgart, Frankfurt: Govi-Verlag GmbH; 1991
min10 20 30 40 50 60
mAU
0
20
40
60
80
100
120
DAD1 A, Sig=260,4 Ref=550,100 (VMT\VT_02042013 2013-04-02 13-12-42\PETASH000132.D)
min10 20 30 40 50 60 70
mAU
0
50
100
150
200
250
300
350
DAD1 B, Sig=325,4 Ref=600,100 (VMT\VT_01042013 2013-04-01 14-14-25\PETASH000106.D)
Figure. 1. HPLC fingerprint of P. hybridus root non-polar extract
recorded on 260 and 325 nm
HPLC chromatogram of the investigated extract poinetd that the P.
hybridus root extract was rich in sesquiterpene lactopnes (Figure 1). The
preliminary investigation indicated phenolic compounds presence, as
well. Namely, moderate phenolic (mg GA/g) and tannins (%) content
were found (29.0 and 0.89, respectively). Flavonoid content was low
(0.1%). Contrary to this, high sesquiterpene lactones content was
revealed (Table 1).