Search

Wormwood herb is regarded as a medicinal raw material rich in bitter principles. However, its biological activity is also affected by the presence of essential oil and phenolic compounds. In Poland wormwood herb is collected mainly from wild growing plants. The aim of the study was to compare the content and composition of essential oil and phenolic compounds in the raw materials obtained from 18 populations of wormwood from the central and north-eastern area of Poland. Herb was collected at the stage of plant blooming and dried naturally. Content of essential oil in the raw materials was determined by hydrodistillation method, and its composition by GC. Methanolic extracts obtained by continuous exhaustive extraction of raw materials were used for qualitative and quantitative analysis of phenolic compounds by HPLC. The content of essential oil in the investigated raw materials ranged from 0.40 to 0.88%. In 6 populations the dominant constituent of essential oil was sabinyl acetate, in three - chrysanthenyl acetate, in two - sabinene, and in one - β-thujone. Other populations were characterised by comparable content of two or three main compounds in essential oil, e.g., sabinene and β-myrcene, β-myrcene and cineol, chrysanthenyl acetate and β-myrcene, sabinyl acetate, chrysanthenyl acetate and β-thujone. In the investigated raw materials 11 phenolic substances were identified: 5 phenolic acids (caffeic, p-coumaric, ferulic, chlorogenic, and rosmarinic one), 5 flavonoids (apigenin, diosmetin, orientin, quercetin, and hyperoside), and (-)-epigallocatechin gallate. In the raw materials obtained from all the investigated populations the dominant phenolic compound was ferulic acid but its content was diverse (396-1118 mg 100 g-1 of dry herb). A remarkable amount of chlorogenic and rosmarinic acids was also found (159-477 and 178-446 mg 100 g-1, respectively).

The aim of the investigation was to compare 10 populations of great burnet growing wild in Poland in respect of phenolic compounds accumulation. Total content of tannins and phenolic acids were determined according to Polish Pharmacopoeia and qualitative analysis by HPLC. District differences in the content of determined phenolic compounds both between the populations originating from different regions of Poland and between the populations from the same region were observed. Differences in the content of particular phenolic compounds between herb and roots were found too. In herb five phenolic acids were identified, namely: chlorogenic, ellagic, gallic, caffeic, and rosmarinic acid, whereas in underground organs two phenolic acids (ellagic and gallic) and four catechin derivatives (/+/-catechin, /-/-epigallocatechin, /-/-epicatechin, and /-/-epigallocatechin gallate). Dominating phenolic compounds in herb were gallic and chlorogenic acids, whereas in underground organs - gallic acid, catechin and epicatechin.

Eleutherococcus senticosus (Rupr. et Maxim.) Maxim. is a thorny shrub, belonging to the Araliaceae family, occurring primarily in Russia, China, Korea and Japan. In traditional medicine underground parts of this plant are used as a drug with adaptogenic activity. Eleutherosides are the most important biologically active compounds responsible for this action. The content of eleutherosides and polyphenolic acids in rhizomes, roots and stem bark was determined to assess the chemical varibility between ten selected E. senticosus clones. HPLC analyses showed significant differences between clones in the content of eleutherosides B and E. The highest content of these compounds was observed in the stem bark (from 207.2 to 566.0 mg 100 g -1 ), lower in the rhizomes (from 47.5 to 326.2 mg 100 g -1 ) and the lowest in roots (from 38.9 to 228.6 mg 100 g -1 ). In all three investigated plant organs presence of chlorogenic, caffeic, rosmarinic, ferulic, protocatechuic acids and rutoside was determined as well.

The aim of this investigation was to determine the genetic and chemical variability of bennet's herb (Geum urbanum) growing wild in the eastern and central areas of Poland. HPLC was applied for determination of phenolic compounds. Two flavan-3-ols and three polyphenolic acids were found in leaves, while five flavan-3-ols and two polyphenolic acids in underground organs. The populations differed especially in respect of the content of catechin and epicatechin in underground organs. The RAPD analysis indicated genetic variability of investigated populations but there was no clear correlation between this variability and the chemical composition of determined phenolic agents.

Common sage (Salvia officinalis L., Lamiaceae family) is a perennial evergreen subshrub native to the Mediterranean. Leaves collected from this plant are used as an important medicinal raw material with antiseptic, antispasmodic, astringent, carminative, cholagogue and tonic activity. The aim of the research was to evaluate the intraspecific variability of sage with special respect to seed setting and germination. The obtained seeds mass fluctuated from 17.0 to 25.5 g per plant. From a single flower 3 seeds were obtained, on average. A clear relationship between the position of seeds in inflorescence and their germinability was observed. The germinability of seeds obtained from upper whorls fluctuated from 92 to 98% whereas germinability of seeds from the lowest one was 70%. There were no distinct differences between plants in the 1000-grain weight. The mass of 1000 grains ranged from 6.090 to 6.161 g.

One of the essential problems in the cultivation of Hypericum perforatum is to obtain healthy and genetically uniform plant material for establishing of plantations. The aim of the study was to compare developmental and chemical characteristics of H. perforatum plants propagated generatively and by micropropagation. The experimental plant material were cv. 'Topas'and two populations (No. 25 and 31) selected in the Department of Vegetable and Medicinal Plants. Micropropagation was carried out using initial shoot explants taken from mature plants growing in the field. Multiple shoot regeneration was obtained on LS (Linsmaier & Skoog) medium supplemented with 3 mg L -1 kinetin, 0.3 mg L -1 indole 3-acetic acid (IAA) and 80 mg L -1 adenine sulfate. Rooting of regenerated shoots was induced on LS (modified LS, containing half of the original concentration of macronutrients) with 0.5 mg L -1 IAA. Both media were supplemented with 20 g L -1 sucrose and 6.5 g L -1 agar. All cultures were incubated at 24°C and 16-hour day. Rooted plants were adapted to ex vitro conditions. The plant material obtained in vitro was compared with that obtained from the seeds sown in the greenhouse. In vitro regenerated and generatively propagated plants did not differ in respect of plant habit. The propagation method did not affect the content of hypericins, polyphenolic acids and flavonoids in the air-dry herb harvested at the full blooming stage. The herb of cv. 'Topas' was characterised by the highest content of hypericins and the lowest content of polyphenolic acids.

The relation between the age of plants and the accumulation of phenolic acids, as well as their composition in particular organs of Rhaponticum carthamoides (Willd.) Iljin [Leuzea carthamoides D.C.] was investigated. The content of phenolic acids in rhizomes was higher than in roots (1.40% and 0.85% respectively). The content of these compounds in basal leaves was higher than in stem leaves. Underground organs and leaves were characterised by different compositions of phenolic acids. The dominant compound in the fraction of free phenolic acids from underground organs (rhizomes and roots) appeared to be gentisic acid and in the fractions of bound acids – o-hydroxyphenylacetic acid. The dominant compound in the fraction of free phenolic acids from basal leaves was protocatechuic acid. The biggest share in the fraction of free phenolic acids from stem leaves had ohydroxyphenylacetic and m-hydroxybenzoic acids. It is worth stressing that both gentisic acid (present in underground organs) and protocatechuic acid (in leaves) are known to reveal an immunotropic activity. INTRODUCTION Rhaponticum carthamoides (Willd.) Iljin [Leuzea carthamoides D.C.] – a perennial originating from Siberia – is considered to be one of the adaptogenic plants. Its rhizomes with roots have been used for many years as a stimulant, roborant and tonic agent. Recently, also Rhaponticum’s leaves have been taken into consideration as a medicinal raw material, especially in respect of their immunotropic properties. So far, it has been stated that immunomodulatory activity of Rhaponticum extracts could be connected with the presence of polysaccharides (Lamer-Zarawska et al., 1996). However, it is known that one of the important groups of plant constituents of immunotropic activity are phenolic compounds, including phenolic acids. They are responsible for many other pharmacological effects, too. Some of them reveal e.g. biligenic, antibacterial and antiviral activity (Borkowski, 1993; Borkowski and Milkowska, 1996). Phenolic acids have not been an object of thorough research in Rhaponticum as yet. The aim of the present study was to determine the content and composition of phenolic acids in underground and above ground organs of Rhaponticum, obtained from plants of different age. MATERIALS AND METHODS Plant Material Plant material for analysis was obtained from the experimental field of Dept. of Vegetable and Medicinal Plants. Rhizomes and roots of one-, twoand three-year-old plants were collected in October. Basal and stem leaves of two-, threeand four-year-old plants were harvested in May (at the stage of plant blooming). The raw materials were dried at 40°C. Quantitative Determination of Phenolic Acids The total polyphenolic acid content (converted into caffeic acid) was determined according to Polish Standard (PN-91/R-87019). Proc. Int. Conf. on MAP Eds. J. Bernath et al. Acta Hort. 597, ISHS 2003 120 Qualitative Determination of Phenolic Acids The isolation of phenolic acid fractions was made according to Krzaczek (1984). Free phenolic acids and those revealed after alkaline and acid hydrolysis were sililated (with BSTFA, Fluck) and identified by a GC method. Gas chromatography was performed on a Hewlett Packard 6890 as follows: capillary column HP-5, length 30 m, diameter 0.32 mm; detector FID temperature 250oC; injector temperature 220oC; carrier gas–helium with the flow rate 2 ml/min. Column temperature was programmed as follows: 60oC (2 min.), temperature increment 8oC/min. and finally 280oC (5 min.). Particular compounds were identified on the ground of retention times of the standard phenolic acids. RESULTS The content of phenolic acids in rhizomes was higher than in roots, regardless of the age of plants (Fig. 1). On the basis of GC, 17 phenolic acids were identified. They were present especially in a free form and some of them also in a bound form. The composition of these compounds in rhizomes and roots of oneand two-year-old plants was similar (in the fractions of free phenolic acids the dominant was gentisic acid and in the fractions of bound acids – o-hydroxyphenylacetic acid). Three-year-old plants were characterised by different composition of phenolic acids. There was no clearly dominant compound in the fractions of free phenolic acids, however in rhizomes a little bigger share in this fraction had protocatechuic acid and in roots p-hydroxybenzoic, protocatechuic and p-coumaric acids. In the fractions obtained after alkaline and acid hydrolysis of extracts, of both rhizomes and roots, the main compound was p-coumaric acid (Table 1). The content of phenolic acids in basal leaves of twoand three-year-old plants was higher than in stem leaves. In four-year-old plants there was no difference in the content of these compounds between basal and stem leaves (Fig. 2). The dominant compound in the fraction of free phenolic acids from basal leaves was protocatechuic acid. In the fractions of bounded acids the main compounds were protocatechuic and phydroxybenzoic acids. In stem leaves, the biggest share in the fraction of free phenolic acid had o-hydroxyphenylacetic and m-hydroxybenzoic acids (Table 2). DISCUSSION Previous studies on Rhaponticum carthamoides dealt with such groups of plant constituents as ecdysteroids (Varga et al., 1986; Girault et al., 1988), flavonoids (Varga et al. 1990), triterpenoid saponins (Vereskovskij et al., 1977) and polyacetylenes (Szendrei et. al., 1984). There was no report on the presence and composition of phenolic acids in Rhaponticum plants. Our preliminary investigation indicated that they are present in considerable amount both in underground and above ground organs of plants. That study showed also differences in the composition of this group of compounds between particular organs of four-year-old plants (Skiba and Weglarz, 1999). The results of the present study confirm that the content of phenolic acids in rhizomes is higher than in roots, with no respect to the age of plants. The composition of this group of plant constituents in rhizomes and roots appeared to be dependent on the age of plants. Threeyear-old plants differed markedly in this respect from oneand two-year-old plants. It is worth emphasising the remarkable dominance of gentisic acid in the fraction of free phenolic acids isolated from rhizomes and roots of oneand two-year-old plants. According to Borkowski (1993), gentisic acid has immunotropic activity – it affects lymphocyte-induced angiogenesis and enhances IgG antibodies production. The other phenolic acid, which seems to be important from the pharmacological point of view, is pcoumaric acid. It is known to have an immunomodulatory and antibacterial activity (Świątek et al., 1984; Borkowski and Milkowska, 1996) and according to our results, it is present in rhizomes and roots of one-, twoand three-year-old plants, both in a free and bound form. The results of our previous and present study indicate that the dominant phenolic

The developmental and chemical differences between H. perforatum, H. maculatum and H. elegans originating in the south of Poland (Krakow district) were investigated. The research was done in oneand two-year-old plantations, respectively, with plants harvested at four stages of development (vegetative phase, before blooming, beginning of blooming and at full blooming). The mass of raw material and the content of flavonoids and hypericins were determined for each harvest. The yield of herb and the content of active compounds were highest in H. elegans. Taking into consideration the mass of herb and the content of investigated compounds, the optimal time for harvest appeared to be the full blooming stage, especially in the second year of vegetation. INTRODUCTION The genus Hypericum comprises a large number of species widely distributed in many countries of Middle Europe, North America and Eastern Africa (Roth, 1990). Of these species, three grow wild in Poland – H. perforatum L., H. maculatum Crantz, and H. elegans Steph. Hypericum maculatum and Hypericum elegans have not yet been used officially in Poland as medicinal raw material, but these species occasionally appear as contamination of H. perforatum collected from natural habitats (Brunarska et al., 1984, Nahrstedt and Butterweck, 1997, Osinska and Weglarz, 2000). The aim of this investigation was to evaluate the phenological and chemical differences among these three species originating in the south area of Poland (Krakow district). MATERIALS AND METHODS The research was done at the experimental field of the Department of Vegetable and Medicinal Plants in 1999, and 2000, in oneand two-year-old plantations established from the seeds collected from natural habitats. The plants were propagated by seeding and subsequent transplanting. During the vegetation period, phenological observations were done. From each species, 25 individual plants were selected at random and harvested at four stages of development (vegetative phase, before blooming, beginning of blooming and full blooming). Dry mass of shoots, the total content of flavonoids (converted into quercetin) by spectrophotometric method (Strzelecka and Kaminska, 1978), and content of hypericins by HPLC (Kowalewski et al., 1981) were determined. RESULTS AND DISCUSSION The results indicate that two species of Hypericum (H. maculatum and H. elegans) growing wild in Poland may be regarded as valuable sources of medicinal raw materials and used equivalently to H. perforatum herb. However, these two species differed from H. perforatum in phenological characteristics. H. elegans plants were characterised by the highest rate of growth and markedly earlier blooming (20 days earlier than H. perforatum and H. maculatum). The highest mass of herb was also observed in H. elegans (Table 1, 2). In the majority of samples, the total content of flavonoids and hypericins was higher in H. elegans and H. maculatum herb than in H. perforatum. During vegetation period, the mass of herb, as well as the content of both Proc. Int. Conf. on MAP Eds. J. Bernath et al. Acta Hort. 576, ISHS 2002 42 investigated groups of compounds, increased to full blooming stage (Fig. 1, 2). The results correspond with the data on H. perforatum obtained by Kordana and Zalecki, (1996) and Jokela and Galambosi (1998). The yield of herb of each species was distinctly higher in the second year of vegetation. The raw materials collected in the second year of vegetation were also characterised by higher content of flavonoids and hypericins. Literature Cited Brunarska, Z., Wegiel, J., Wiatr, E., Ekiert, H. and Kohlmunzer, S. 1984. St. John’s wort, Hypericum perforatum L., a valuable pharmaceutical raw material and the chances for the protection of its natural resources. Studia Naturae-Seria A 24:51-66. Jokela, K. and Galambosi, B. 1998. Effect of ridge cultivation and plant density on growth and yield of some medicinal plants in Finland. Z. Arzn. Gew. pfl. 3:139-145. Kordana, S. and Zalecki, R. 1996. The researches on cultivation of Hypericum perforatum L. Herba Pol. 3:144–150. Kowalewski, Z., Kortus, M. and Tyczynski, T. 1981. Determination of hypericin in the Hypericum perforatum L. herb and its formulations. Herba Pol. 4:295-302. Nahrstedt, A. and Butterweck, V. 1997. Biologically active and other chemical constituents of the herb of Hypericum perforatum L. Pharmacopsychiat. 30. (Suppl.):129-134. Osinska, E. and Weglarz, Z. 2000. Preliminary investigation on variability of Hypericum perforatum L. in Poland. Abstr. Intl. Congr. and 48 Annu. Meeting of the Society for Medicinal Plant Res. Zurich, Switzerland, 3-7 September. Roth, L. 1990. Botanik von Hypericum. Ecomed. Landsberg. 15–87. Strzelecka, H. and Kaminska J. 1978. Chemiczne metody badan roślinnych surowcow zielarskich. PZWL, Warszawa, 49-51.