Chemistry for Sustainable Development 21 (2013) 369374 369 UDC 633.8+615.322 Biologically Active Substances in Filipendula ulmaria (L.) Maxim. Growing in the Middle Urals G. I. VYSOCHINA1, T. A. KUKUSHKINA1 and E. S. VASFILOV2 1Central Siberian Botanical Garden, Siberian Branch of the Russian Academy of Sciences, Ul. Zolotodolinskaya 101, Novosibirsk 630090 (Russia) E-mail: [email protected] 2Botanical Garden, Ural Branch of the Russian Academy of Sciences, Ul. 8 Marta 202a, Yekaterinburg 620144 (Russia) (Received March 18, 2013; revised May 24, 2013) Abstract The plants of Filipendula ulmaria (L.) Maxim. growing in the Middle Urals can serve as a source of flavonols, catechins, tannins, saponins and carotenoids. A definite specificity in their accumulation in or- gans was established. The high carotenoid content in the leaves of Filipendula ulmaria was established by us for the first time. Key words: Filipendula ulmaria (L.) Maxim., flavonols, catechins, tannins, pectic substances, saponins, carotenoids INTRODUCTION tory and wound-healing medicine [7]. The Fil- ipendula ulmaria raw material basis is rather Much attention is attracted during the re- well provided by natural resources [8]. cent years to the development of the prepara- Meadowsweet is used in folk medicine to treat tions of plant origin. Special interest is caused various diseases; all parts of the plant are used in this respect by the plants that are widely (flowers, leaves, roots) in the form of decoc- used in folk medicine in different countries. tions, water extracts, tinctures, and ointments. Meadowsweet Filipendula ulmaria (L.) Maxim. It is often recommended to use separate organs (Rosaceae fam.) is used in folk medicine in Russia or their combinations, which is undoubtedly due and in some European countries as a corrobo- to the differences in the chemical composition rant, anti-inflammatory, wound-healing, an- of active substances present in different parts algesic, diuretic, antiulcerogenic, hypoglycemic, of plants [8]. Preparations of meadowsweet flow- sedative, antihemorrhoid and astringent means ers have won the widest application in folk med- [1, 2]. At present, enormous data are also avail- icine [9]. Water extract of its flowers is used to able on other kinds of pharmacological activity treat rheumatism, asthma [10, 11], stomach and (stress-protective, immunomodulating, antitu- duodenal ulcer [1214], water extract of leaves mour). Special attention of researchers is paid is used to treat trophic ulcer of lower extremi- to the possibility to use meadowsweet raw ma- ties, wounds and burns [13, 15, 16]; decoction terial to develop therapeutic and prophylactic of roots is applied to treat nervous disorders, preparations with high antioxidant activity [3 hypertension, as antihelminthic agent and spas- 6]. The flowers of Filipendula ulmaria are used molytics [11, 13]. Herb in general also possesses in Russian scientific medicine as anti-inflamma- valuable pharmacological properties [17, 18]. Vysochina G. I., Kukushkina T. A. and Vasfilov E. S., 2013 370 G. I. VYSOCHINA et al. Wide application of Filipendula ulmaria to down the growth of tumours and accelerate treat various pathologies is connected with the wound healing [22]. diverse chemical composition of the plant; it is The goal of the work was to study the fea- very interesting to study its chemical tures of the accumulation of various groups composition. According to the data of [8, 19, of biologically active substances (BAS) in sep- 20], Filipendula ulmaria plants contain phenol arate organs of Filipendula ulmaria. compounds (flavonols, catechins and tannins). These compounds play important part in various physiological processes involving redox reactions. MATERIALS AND METHODS Preparations based on these compounds are used as bactericidal, antiseptic, anti-inflammatory, The object of investigation was mead- haemostatic means. They are able to inhibit owsweet F. ulmaria (L.) Maxim. (Rosaceae fam- chain free radical reactions, which determines ily) a large (40100 cm high) perennial her- their efficiency in chemotherapy of cancer. baceous plant with short horizontal rhizome. The Pectic substances present in meadowsweet leaves are green from above and white-pan- leaves carry out detoxication of human organ- nose from beneath. Inflorescences are panicu- ism by binding and removing toxins, poisons and late and large. Flowers are 78 mm in diame- radioactive isotopes, which is especially impor- ter, their colour is yellowing-white. This is Eur- tant [21]. Due to the presence of saponins, Fil- asian species. Its areal embraces almost the ipendula ulmaria is efficient in treating vascular whole territory of Russia and partially the ter- sclerosis, atherosclerosis combined with hyper- ritories of Central and Northern Europe. Mead- tension, and malignant neoplasms [20]. The im- owsweet grows on floodplain meadows, grass portance of carotenoids which were detected by bogs, along river banks, in humid forests, cu- us in the leaves of Filipendula ulmaria for the tover patches [23]. first time cannot be overestimated. Undergoing The raw material of meadowsweet was col- transformation into vitamin A, they affect en- lected in 20102012 in different natural popu- docrine and reproductive system, ensure the sta- lations at the territory of Sverdlovsk Region bility of organism to infections and mycoses, slow (the Middle Urals). We also analyzed the raw TABLE 1 Characterization of the samples of raw material of Filipendula ulmaria (L.) Maxim. Sample Description of sample collection site Year No. of sample collection Natural populations 1 Kamyshlov District, near Obukhovo village; the valley of the Pyshma River, along the field path. 2010 Relatively dry, well illuminated place 2 Achit District, near Yalym settlement; brook bank, in a low place near the motorway. Wet, well 2011 iluminated place 3 To the south from Revda torn, the bank of the Bolshaya Puzanikha River. Well illuminated wetland. 2012 4 Territory related to Polevskiy town, near Kurganovo village, low place near forest road. 2012 Rather wet, weakly illuminated place 5 Pyshma District, near Cheremysh settlement, forest edge not far from brook bank. Relatively dry, 2012 well illuminated place 6 Alapayevsk District, near Aromashevo village, in a low place near the motorway. Rather wet 2012 well illuminated place 7 Territory related to Berezovskoye town, near Klyuchevsk settlement, forest edge, along motorway. 2012 Wet, well illuminated place Culture conditions 8 Botanical Garden, Ural Branch of the RAS, Yekaterinburg. Dry, well illuminated place 2011 BIOLOGICALLY ACTIVE SUBSTANCES IN FILIPENDULA ULMARIA (L.) MAXIM. 371 material of F. ulmaria cultivated in the Botan- hydrochloric acid was added to each sample; ical Garden, Ural Branch of the RAS, Yekat- 4 mL of concentrated hydrochloric acid were erinburg (Table 1). Flowers, leaves and stems added to reference tubes. The volume was brought were stores in July during mass blossoming of up to 5 mL in each tube. After 5 min, in the case the plants. The sample for biochemical investi- of the presence of catechins, the colour became gation was composed of several ten individuals rose. The concentration of catechins in the sam- from each population. ples was determined from the calibration curve To determine the concentrations of flavonols, plotted for (±)-catechin Sigma [25]. catechins, tannins, pectic substances, saponins, The content of tannins was determined by carotenoids, we used raw material dried in shad- means of titration [26]. A precise weighted por- ow in aired rooms. All biochemical indices were tion of air-dry raw material (0.51 g) was ex- calculated per mass of absolutely dry raw ma- tracted by water on boiling water bath for 45 terial. The result was averaged over three par- min. The resulting extract 510 mL in volume allel determinations for each parameter. was titrated with a 0.1 N KMnO4 solution in a Quantitative determination of flavonols was beaker with 400 mL of water in the presence based on the method proposed by V. V. Belikov of indigo carmine under permanent mixing till and M. S. Shraiber [24]. It involves the forma- golden-yellow colour. In parallel, reference ti- tion of the complex of flavonols with alumin- tration without the extract under study was ium chloride. The precise weighted portion of carried out. Calculated for tannin, 1 mL of ground raw material (about 0.5 g) was placed 0.1 N KMnO4 solution is equivalent to 4.157 mg in a flask 100 mL in volume, and complete ex- of tanning substances. traction with 70 % ethanol on the boiling wa- Pectic substances (protopectins and pectins) ter bath was carried out. The completeness of were determined using the carbazole method extraction was monitored with the help of the based on obtaining specific violet-rose colour of reaction with 5 % NaOH solution. Then 0.1 mL uronic acids with carbazole in sulphuric medi- was placed in a volumetric tube, 0.2 mL of 2 % um. The density of coloured solutions was mea- AlCl3 solution in 96 % ethanol was added, and sured with the help of a FEK-56M photoelec- the volume was brought up to 5 ml using etha- tric colorimeter at the wavelength of 535 nm nol of the same concentration. In the reference in a cell with the working length of 5 mm. The version, 12 drops of 30 % acetic acid were content of pectic substances was determined added to 0.1 mL of the extract and then the using the calibration curve plotted with galac- volume was brought up to 5 mL. The solutions turonic acid [27]. were mixed; optical density of the solution with Saponin content was determined using the aluminium chloride was measured after 40 min gravimetric method. Approximately 2 g of the with a SF-26 spectrophotometer at 415 nm in a air-dry material was extracted with chloroform cell with the layer thickness 1 cm using a cell in Soxhlet extractor till complete decolouring with the acid as a reference. to remove lipids and gum which hinder sapo- Total content of flavonols (in per cent of nin determination.