Received: 4 January, 2008. Accepted: 17 February, 2008.
Short Communication
Medicinal and Aromatic Plant Science and Biotechnology ©2008 Global Science Books
Detection of Hyperforin in Turkish Species of Hypericum
(Guttiferae)
Ali Kemal Ayan
1
• Cüneyt Çrak
2*
1
The High School of Profession of Bafra, University of Ondokuz Mays, Samsun, Turkey
2
Faculty of Agriculture, Department of Agronomy, University of Ondokuz Mays, Kurupelit, Samsun, Turkey
Corresponding author: * cuneytc@omu.edu.tr
ABSTRACT
In the present study, six Hypericum species from the Turkish flora were investigated for the presence of hyperforin namely H.
heterophyllum Vent, H. hyssopifolium L., H. linarioides Bosse, H. orientale L., H. scabrum L. and H. triquetrifolium Turra. For this
purpose, the aerial parts were collected at full flowering, dissected into floral, leaf and stem tissues, air-dried at room temperature and then
assayed for hyperforin by HPLC. Hyperforin was detected only in flower tissues of H. hyssopifolium (29.2 mg/g DW) and H. linarioides
(6.28 mg/g DW). This data could be useful for elucidation of the chemotaxonomical significance of hyperforin and for the phytochemical
evaluation of H. hyssopifolium and H. linarioides.
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Keywords: HPLC, Hypericum hyssopifolium, Hypericum linarioides
INTRODUCTION
Hypericum is a large genus of herbs or shrubs which grow
in temperate regions of the world and the species belonging
to this genus have been used as traditional medicinal plants
due to their various medicinal properties for hundred of
years (Demirci et al. 2005). In particular, extracts of Hype-
ricum perforatum L. are now widely used in Europe as a
drug for the treatment of depression (Patocka 2003). The
Hypericum genus of Guttiferae is represented in Turkey by
89 species of which 43 are endemic (Davis 1988).
Hypericum plants have been reported to contain many
bioactive secondary metabolites from different classes
namely naphthodianthrones, phloroglucinols, flavonoids,
phenylpropanes, essential oils, amino acids, xanthones,
tannins, procyanidins and other water-soluble components
which possess a wide array of biological properties (Gree-
son et al. 2001; Kitanov 2001; Çrak et al. 2006; Tanaka
and Takaishi 2006).
Many pharmacological activities of Hypericum extracts
appear to be attributable to their hypericins and hyperforin
content (Barnes et al. 2001). Results from recent studies
have indicated hyperforin as the main chemical, responsible
for antidepressant effects of Hypericum extracts (Roz and
Rehavi 2004). It also exhibits anti-inflammatory (Feisst and
Werz 2004), antitumoral (Schwarz et al. 2003) and antian-
giogenic (Dona et al. 2004) effects. It has been recommen-
ded as a marker compound for the routine standardization
of Hypericum products (Gerlie and Koda 2001). Due to
these reasons, many species of Hypericum have been inves-
tigated for the presence of hyperforin, but only a few of
them were reported to contain this compound (Kirakosyan
et al. 2003; Maggi et al. 2004; Piovan et al. 2004; Klingauf
et al. 2005; Martonfi et al. 2006; Smelcerovic et al. 2006).
In the present study, the aim was to determine hyper-
forin content in stems, leaves and flowers of some Hype-
ricum species growing wild in Turkey namely H. hetero-
phyllum Vent, H. hyssopifolium L., H. linarioides Bosse, H.
orientale L., H. scabrum L. and H. triquetrifolium Turra.
MATERIALS AND METHODS
Plant material
The aerial parts of the aforesaid Hypericum plants were collected
at full flowering in June, 2005 from three sites of Northern Tur-
key: Maçka (40 49 N; 39 37 E; 270 m sea level), Erbaa (40 41
N; 36 34 E; 230 m sea level) and Kastamonu (41 24 N; 33 45
E; 790 m sea level) and identified by Dr. Hasan Korkmaz, Depart-
ment of Biology, University of 19 Mayis, Samsun-Turkey. Vou-
cher specimens were deposited in the herbarium of Ondokuz
Mayis University Agricultural Faculty (OMUZF # 127-H. hetero-
phyllum, OMUZF #128-H. hyssopifolium, OMUZF # 129-H. lina-
rioides, OMUZF #131-H. orientale, OMUZF #133-H. scabrum
and OMUZF#134-H. triquetrifolium). The plant materials were
dissected into floral, leaf and stem tissues, air-dried at room tem-
perature and grounded to powder using a laboratory mill, then
assayed for hyperforin by HPLC.
Chemicals
Reference standard of hyperforin was purchased from ChromaDex,
Inc. (Laguna Hills, CA, USA). The high performance liquid chro-
matography (HPLC)-grade acetonitrile, acetone and methanol
were purchased form Caledon (Mississauga, ON, Canada). Tri-
ethylammonium acetate is a product of Sigma-Aldrich Canada
(Oakville, ON, Canada).
Extraction and High Performance Liquid
Chromatography (HPLC) analysis of hyperforin
The isolation and analysis of hyperforin were done according to
protocols reported by Murch et al. (2002). About 100 mg sample
was transferred into an amber-colored 20 ml vial with 5 ml ace-
tone:methanol (50:50, v:v) and sonicated for 30 min (Ultra-sonic
FS-14 Sonicator; Fisher Scientific, Nepean, ON, Canada). The
sample was centrifuged at 3000 rpm for 10 min (GS-6 series cen-
trifuge, Beckman Instruments Inc, Palo Alto, CA, USA) and the
supernatant was filtered using 0.2 μm nylon syringe filter (Waters
Chromatography Inc., Mississauga, ON, Canada). Aliquots of each
sample (500 μl) were transferred into a clear glass auto-sampler
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