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Composition and Biological Activity of Rhaponticum Carthamoides Extracts Obtained from Plants Collected in Poland and Russia

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Composition and Biological Activity of Rhaponticum Carthamoides Extracts Obtained from Plants Collected in Poland and Russia Journal of Medicinal Plants Research Vol. 7(11), pp. 687-695, 17 March, 2013 Available online at http://www.academicjournals.org/JMPR DOI:10.5897/JMPR012.1061 ISSN 1996-0875 ©2013 Academic Journals Full Length Research Paper Composition and biological activity of Rhaponticum carthamoides extracts obtained from plants collected in Poland and Russia Edyta Biskup1,3*, Berenika Szynklarz1, Marek Golebiowski2, Kamil Borsuk2, Piotr Stepnowski2 and Ewa Lojkowska2 1Department of Plant Protection and Biotechnology, Intercollegiate Faculty of Biotechnology University of Gdansk & Medical University of Gdansk, Kladki 24, 80-822 Gdansk, Poland. 2Department of Environmental Analysis, Faculty of Chemistry, University of Gdansk, Sobieskiego 18/19, 80-952 Gdansk, Poland. 3Department of Dermatology, Bispebjerg Hospital, Bispebjerg Bakke 23, 2400 Copenhagen, Denmark. Accepted 10 December, 2012 Rhaponticum carthamoides is a medicinal plant indigenous to Siberia. Although its root tissue is a well- known source of biologically active compounds, recent studies have focused on the above-ground part of the plant due to its easier accessibility. An R. carthamoides plantation was recently established in Northern Poland. Composition and antioxidant activity were compared for R. carthamoides leaf extracts obtained from plants grown in two plantations: in Poland (Lubiewice) and in Russia (Koriazma); the latter plantation has growing conditions similar to those in the R. carthamoides natural habitat. Plant secondary compounds were extracted with chloroform, methanol or water. As indicated by gas chromatography-mass spectrometry (GC-MS), the content of most compounds analyzed was higher in extracts from plants grown in Lubiewice (Poland). Antioxidative activity (as measured by diphenyl 1- picrylhydrazyl free radical (DPPH) and ferric reducing antioxidant power (FRAP) assays) and cytotoxic activity (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay) were higher in extracts from the Polish material. Key words: Antioxidant activity, cytotoxic activity, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), diphenyl 1-picrylhydrazyl free radical (DPPH), ferric reducing antioxidant power (FRAP), gas chromatography-mass spectrometry (GC-MS). INTRODUCTION The number of known plant-derived active substances same time the cosmetics and pharmaceutical industries has already surpassed 50,000 (Kohlmünzer, 2000). require raw material of high value and standardized Importance of these compounds as components of composition. However, plant biochemical profiles may cosmetics and pharmaceuticals is growing, though in the differ, depending on the place of origin due to various climate, photoperiod and soil conditions (Castro et al., 2001; Szentmihályi et al., 2004). Rhaponticum carthamoides (Willd) Iljin (synonyms: *Corresponding author. E-mail: [email protected]. Tel: Leuzea carthamoides, maral’s root) has a long story of +45 5031 8853. Fax: +45 3531 6010. use in Siberian folk medicine, where its roots, already in the times of first Russian settlers, were applied to treat Abbreviations: DW, dry weight; DPPH, diphenyl 1- picrylhydrazyl free radical; FRAP, ferric reducing antioxidant overstrain and common weakness after illnesses (Yance, power; GC-MS, gas chromatography-mass spectrometry; MTT, 2004). Systematic scientific studies of R. carthamoides 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide. extracts (RCEs) activity and composition, verifying the 688 J. Med. Plants Res. traditional use, began in 1940-ties, mainly in the former interesting also from the commercial point of view. Soviet Union and Japan. Petkov et al. (1984) reported Currently, a number of preparations based on that water-ethanol extract from R. carthamoides roots R. carthamoides-derived compounds, mainly designed increased rats physical performance (forced-swim test) for sportsmen, are commercially available. An R. and mice learning ability (maze test). Todorov et al. carthamoides plantation was recently established in (2000) observed anabolic effect of the root RCE in mice Lubiewice, Poland, with the intention of producing R. and ascribed it to the presence of 20-hydroxyecdysone. carthamoides preparations for the Polish market Recently, Maslov and Guzarova (2007) described (FITOSTAR, personal information, cardioprotective and anti-arrhythmic effect of RCE. http://fitostar,pl/preparat_rapontik). Along with the research, related to the physiological This study presents a preliminary comparison of the effects, the analysis of plant extracts composition was composition of extracts isolated, from R. carthamoides being run. Initially, R. carthamoides gained attention as a plants growing under conditions similar to those in their rich source of ecdysteroids, compounds with potentially natural habitat (Russian Federation) and from R. adaptogenic and anabolic activity (Yance, 2004). 20- carthamoides growing in the plantation recently hydroxyecdyson is the main R. carthamoides ecdys- established in Lubiewice, Poland. teroid, however, almost 50 others have been isolated so We decided to concentrate on the R. carthamoides far (Kokoska and Janovska, 2009), of which 17 were above-ground parts (leaf extracts), though traditionally characterized only during the last 4 years (Budesinky et roots and rhizomes constituted the pharmaceutical raw al., 2008). material. However, it should be underlined, that this In addition to ecdysteroids, several other biologically approach means eliminating the plant and precluding the active compounds have been isolated from its above- possibility of annual harvest, without renewing the and underground organs. Over 30 terpenoids have been plantation. There is also a growing number of records identified in volatile oils (here: sesquiterpenes constituting demonstrating high biological activities of constituents nearly 70% of the volume) obtained from R. obtained from R. carhamoides herb, mainly antioxidative carthamoides roots and rhizomes (Havlik et al., 2009). and immunomodulatory (Kokoska and Janovska, 2009) Additionally, the underground part contains phytosterols, and pointing to the above ground part as a more triterpenoid alcohol (carthamenyl) (Grimshaw et al., 1981) abundant source of polyphenolic constituents, than roots and thiophene polyacetylenes exhibiting antifungal and rhizomes (Skiba and Weglarz, 1999). Therefore, (Chobot et al., 2003) and phototoxic (Chobot et al., 2006) using the R. carthamoides aerial parts is scientifically activity. In R. carthamoides seeds, N-feruloserotonins warranted and might be interesting from the economical were detected (Pavlik et al., 2002), compounds showing point of view. mild antinoceptive activity (Yamamotová et al., 2007). A range of polyphenols and phenolic acids were also indentified in R. carthamoides underground tissue, e.g. MATERIALS AND METHODS stilbene derivative (Hajdu et al., 1998), gentisic acid, o- hydroxyphenylacetic acid (Skiba and Weglarz, 2003), Plant Material accompanied by a number of flavonoid glycosides, Plant material, supplied in the form of R. carthamoides air-dried mainly the derivatives of quercetin and isorhamnetin powdered leaves, was a generous gift from Professor N. P. (Sharaf et al., 2001; Varga et al., 1990). Timofeev and FITOSTAR™. Leaves were obtained from plants Flavonoid glycosides (mainly containing quercetin, grown in two locations. patuletin and campherol as aglycone part) were also 1) Koriazma (Arkhangelsk region, Russian Federation): leaves were found in R. carthamoides herb (Koleckar et al., 2010; harvested in the summer of 2003 and 2006. Samples collected at Miliauskas et al., 2005; Varga et al., 1990), together with different growth stages were pooled together. Further in the text are phenol acids-protocatechuic (Skiba and Węglarz, 2003) referred to as the “K-rus” material. and chlorogenic (Miliauskas et al., 2005). Polyphenols 2) Lubiewice (Kujawsko-Pomorskie voivodeship, Republic of (mainly flavonoid glycosides) are believed to be the class Poland): material was collected every 5 to 7 days during the of compounds responsible for high antioxidative and anti- summer of 2006 (soil class IV and V; 18 samples in total) and 2007 (flowering and vegetative shoots; 6 samples in total). Further in the free-radical potential of RCEs, described by Miliauskas et text referred to as the “L-pol” material. al. (2005). On the other hand the ability of methanol RCE to inhibit the platelet aggregation, induced by adenosine Detailed description of climate and photoperiod data of both diphosphate (ADP) or collagen, should be ascribed to locations is as shown in Table 1. free flavonoids forms (Koleckar et al., 2008). Flavonoids and caffeic acid derivatives seem to be also responsible for the RCEs inhibitory effect towards NK-κB (Peschel et Soil class definitions al., 2011). Class IV soils (the average arable land): yields of crops grown on Complex and rich chemical composition as well as a these soils are significantly lower than the soils of the upper. These wide range of activities exhibited, makes R. carhamoides soils are very susceptible to fluctuations in groundwater levels. Biskup et al. 689 Table 1. Comparison of the climate conditions in Lubiewice (Republic of Poland) and Koriazma (Russian Federation). Site of sample collection Koriazma Lubiewice Meteorological station location Kotlas Chojnice and Sliwice 61.32ºN 53.47ºN Coordinates of the sample collection site 47.15ºE 18.07ºE Climate type Temperate continental (subarctic) Temperate continental (warm)
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