Erodium Cicutarium L
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Notes 881 Antioxidative Effect of Extracts from synthesis of prostaglandins, thromboxanes and Erodium cicutarium L. leukotrienes which are responsible for a number Zbigniew Sroka, Halina Rządkowska-Bodalska of physiological processes such as immunity, blood and Irena Mażol pressure and platelet aggregation (Halliwell and Gutteridge, 1988 a). Department of Pharmacognosy, Medical Academy of Wroclaw, PI. Nankiera 1, 50-140 Wroclaw, Poland Another group of free radical reactions are processes caused among others by UV radiation, Z. Naturforsch. 49c, 881-884 (1994); received March 3/June 22, 1994 metal cations (Fe2+, Cu2+) or abnormal products Erodium cicutarium (Storksbill, Hemlock, Storkshill) of porphyrin metabolism. Disorders of the free Extracts, Antioxidative Activity, Oenothera paradoxa radical control system lead to an concentration in (Evening Primrose) Oil crease of very active free radicals such as OH' Extracts from Erodium cicutarium L. (EC) and some (Halliwell and Gutteridge, 1988b), which initiate standard substances present in EC extracts were tested processes leading, among others, to a modification for their antioxidative properties during Fe2+-induced triglyceride oxidation. Hydrophobic fractions such as of DNA or lipid peroxidation. Oxidation of lipids petroleum ether (PEE), benzene (BE) and chloroform disrupts biological natural membranes which may I (ChE I) extracts as well as hydrophilic fractions i.e. result in pathological effects. water (WE) and ethyl acetate (EAE) showed an antioxi In this study the influence of extracts from Ero dative effect. Among the tested substances vitamin C and some polyphenolic compounds; tannin, (+)-catechin dium cicutarium on oxidation of Oenothera para and gallic acid exhibited antioxidative activity, stronger doxa (OP) oil, in the presence of Fe2+ was in than the one of the mentioned extracts. vestigated. Materials and Methods Introduction Preparation o f extracts Recently, much attention has been paid to bio logical properties of various plant extracts and The raw material (the dried Erodium cicutarium their application in therapy. They enhance anti herb, collected from its natural habitat in the body production (Kumazawa et al., 1982; Kuma- northern part of Poland in autumn) was subjected zawa et al., 1985), stimulate macrophages (Pace to fractionated extraction in the Soxhlet apparatus et al., 1983), B lymphocytes or NK cells (Kuma with the following solvents, supplied successively: zawa et al., 1984) and also increase interferon syn petroleum ether (PEE), benzene (BE), chloro thesis (Zieliriska-Jenczylik et al., 1988). form (ChE I) and methanol (ME). From the ob Erodium cicutarium (EC) is a one- or two-year tained extracts the solvent was distilled off and the plant found in the fields, roadsides of the lowlands remaining residue was dried. After concentration and lower piedmonts in Poland. It is known for its of the ME extract the residue was mixed with antihemorrhagic activity, antiviral effect in rela 10 parts of boiling water, and heated of 100 °C tion to myxoviruses, Herpes virus type 1, vesicular (water bath) for 2 h. The cooled water solution stomatitis and vaccinia virus (Zieliriska-Jenczylik was separated from the formed sticky precipitate. et al., 1987). The clear filtrate (WE) was exhaustively extracted Reactions producing free radicals in the human with chloroform, then with ethyl acetate and fi body take place very often. Some of them such as nally with ethyl ether. The obtained three frac unsaturated fatty acids oxidation are very impor tions: chloroform (ChE II), ethyl acetate (EAE) tant in natural physiological processes leading to and ethyl ether (EEE) were evaporated under re duced pressure and dried. All the fractions: PEE, BE, ChE I, WE, ChE II, EAE, EEE and the water residue after extraction Reprint requests to Dr. Z. Sroka. process (WR) were tested for their antioxidative Telefax: 004871442277. activity simultaneously with the standard sub- 0939-5075/94/1100-0881 $ 06.00 © 1994 Verlag der Zeitschrift für Naturforschung. All rights reserved. 882 Notes stances identified previously in Erodium cicuta- in the presence of the investigated compounds rium herb. was calculated according to the equation shown below. Measurement of OP oil oxidation [igC - [igP 100 = / Emulsion: 0.24% (v/v) OP oil in 3.5% (w/v) of ^gC arabic gum in 0.2 m Tris(2-amino-2-hydroxy- methyl-l,3-propanediol)-HCl buffer, pH 7.4, was |igC, |ig of malonaldehyde formed in control test; shaken vigorously for 1 0 min. [igP, |ig of malonaldehyde measured in sample TBA reagent: 0.375 g of thiobarbituric acid after tested compound addition; /, percentage of (TBA) was dissolved (boiling bath) in 30 ml H 2 0, inhibition. then 15 g of trichloroacetic acid and 2.1 ml of 10 n HC1 was added. The solution was adjusted to 1 0 0 ml with distilled water. Methods: The antioxidant activities of extracts Table I. The influence of various extracts from Erodium isolated from Erodium cicutarium were measured cicutarium on the oxidation of OP oil. The rate of the according to the method of Buege and Aust (1978) reaction was expressed as |ig of malonaldehyde formed with some modifications. in the sample after 30, 60, 120 and 180 min incubation. When antioxidative effect of WE, ChE II, EAE, Type of extract Time of reaction [min] EEE, WR and standard substances were studied 30 60 120 180 the following procedure was applied. 1, 2 or 5 mg Malonaldehyde formed in the reaction mixture [[xg] of examined substances were added into 1 2 ml of emulsion. Control tests were prepared without ef Petroleum ether control 0.90 1.12 1.85 2.41 fectors. After 5 min preincubation at 25 °C, 100 |xl extract (PEE) 1 mg 0.22 0.56 0.90 1.57 2 mg 0.34 0.34 0.78 1.01 of 37.8 mM of freshly prepared Mohr salt 5 mg 0.17 0.06 0.22 0.50 ((NH 4 )2 S0 4 xFeS 0 4 x 6 H 2 0 ) solution in water Benzene extract control 1.06 1.51 2.36 3.03 was added into the mixture. 0.5 ml of emulsion (BE) 1 mg 0.56 0.95 1.68 1.91 was sampled and moved into 2 ml of TBA solu 2 mg 0.34 0.78 1.51 1.68 5 mg 0.45 0.73 1.23 1.23 tion, heated (boiling bath) for 2 0 min, and centri Chloroform ex control 0.95 1.51 2.24 2.86 fuged for 10 min at lOOOxg. Sample absorption tract I (ChE I) 1 mg 0.00 0.56 0.95 1.12 was measured at 535 nm in a 1 cm glass cuvette. 2 mg 0.00 0.56 0.90 1.01 5 mg 0.00 0.11 0.22 0.50 The amount of oxidation product was deter Water extract control 1.40 1.57 2.13 2.69 mined after 0, 30, 60, 120, 180 min of incubation (WE) 2 mg 0.92 1.04 1.15 1.37 with ferrous ions and expressed as |ig of malon- 5 mg 0.17 0.00 0.11 0.22 Chloroform ex control 1.35 1.79 2.58 3.42 aldehyde formed in a 1 2 ml reaction mixture. tract II (ChE II) 1 mg 1.23 1.40 2.24 2.92 When the influence of hydrophobic fractions 2 mg 1.12 1.35 2.13 2.52 PEE, BE and ChE I on oxidation OP oil was 5 mg 1.06 1.18 1.46 1.63 determined the following changes in measure Ethyl acetate control 1.40 1.57 2.47 3.20 extract (EAE) 1 mg 1.12 1.29 1.85 2.13 ment procedure were introduced. 1,2 or 5 mg of 2 mg 1.12 1.12 1.68 1.57 the extract to be assayed were dissolved in 2 ml 5 mg 0.22 0.11 0.22 0.00 Ethyl ether control 1.12 1.68 2.02 2.52 of chloroform. Then, 0.5 ml of 6 % (v/v) solution extract (EEE) 1 mg 1.18 1.63 1.91 2.75 of OP oil in chloroform was added. After stir 2 mg 1.12 1.63 1.85 2.24 ring, the sample was evaporated under reduced Water residue control 0.90 1.29 1.68 2.36 pressure in the presence of nitrogen. 1 2 ml of (WR) 1 mg 0.90 1.12 2.13 2.24 2 mg 0.78 1.23 1.68 2.02 3.5% (w/v) of arabic gum solution in 0.2 m Tris- 5 mg 0.73 1.12 1.63 2.36 HCl buffer, pH 7.4, was added and the emulsion was prepared as previously described. Further ChE I, chloroform extract obtained from raw material steps in the measurement of the oxidation proc (see preparation of extracts - section Materials and Methods). ChE II. chloroform extract obtained from hot ess were carried out according to the method water fraction (see preparation of extracts - section described above. The inhibition of the reaction Materials and Methods). Notes 883 Results and Discussion 120 and 180 min. When 5 mg of WE or EAE were The malonaldehyde formed during OP oil added the inhibition was similar to the effect ob oxidation and the reaction inhibition in the served with 5 mg of PEE and ChE I. ChE II, EEE presence of EC extracts is shown in Tables I and WR had the lowest effect on oxidation of and II. OP oil. PEE and ChE I exhibited similar effect on oxi Tannin, catechins, gallic and elagic acids, sugars dation of OP oil. In the presence of 1 mg of PEE (glucose, galactose, fructose) amino acids (glycine, in the sample inhibition of malonaldehyde forma alanine, proline, histidine, tryptophan, tyrosine, tion was equal to 76, 50, 51, 35% of the control glutamic acid), vitamins K and C were identified (without extract) after 30, 60, 120 and 180 min of in Erodium cicutarium extracts.