Antioxidant and Antihaemolytic Activities of Ferula Foetida Regel (Umbelliferae)

European Review for Medical and Pharmacological Sciences 2011; 15: 157-164 Antioxidant and antihaemolytic activities of Ferula foetida regel (Umbelliferae)

S.M. NABAVI1, M.A. EBRAHIMZADEH1*, S.F. NABAVI1, B. ESLAMI2, A.A. DEHPOUR2

1Pharmaceutical Sciences Research Center, School of Pharmacy and Traditional and Complementary Medicine Research Center, Mazandaran University of Medical Sciences, Sari (Iran) 2Department of Biology, Islamic Azad University, Qaemshahr Branch, Qaemshahr (Iran)

Abstract. – Objectives: The Ferula genus Introduction (Umbelliferae) is a rich source of gum-resin and is much utilized in folklore medicine. This study It is commonly accepted that under situations is designed to examine antioxidant and anti- of oxidative stress, reactive oxygen species haemolytic activities of Ferula foetida regel flower, stem and leaf extracts. (ROS) such as superoxide, hydroxyl and peroxyl Materials and Methods: 1,1-Diphenyl-2- radicals are generated. The ROS play an impor- picryl hydrazyl radical (DPPH), nitric oxide and tant role related to degenerative or pathological 2+ H2O2 scavenging activities, Fe chelating ability, processes such as aging, cancer, coronary heart reducing power and hemoglobin-induced linole- disease, Alzheimer’s disease, neurodegenerative ic acid peroxidation were used to evaluate an- disorders, atherosclerosis, cataracts, and inflam- tioxidant activities. Antihaemolytic activity was mation1,2. Minimizing oxidative damage may evaluated by H2O2 induced hemolysis in rat erythrocyte. Total phenolic compounds were deter- well be one of the most important approaches to mined as gallic acid equivalents and total the primary prevention of these aging-associated flavonoid contents were calculated as quercetin diseases ad health problems, since antioxidants equivalents from a calibration curve. terminate direct ROS attacks and radical-mediat- Results: The leaf aqueous-ethanol extract ed oxidative reactions, and appear to be of prima- showed the highest activity in DPPH radical ry importance in the prevention of these diseases scavenging activity. All extracts showed weak 3 nitric oxide scavenging activity. The stem ex- and health problems . The human body has sev- tract had better activity in nitric oxide scaveng- eral mechanisms to counteract oxidative stress by ing model than the other extracts (IC50 = 896.9 ± producing antioxidants, which are either natural- 21.9 µg ml-1), but it was not comparable to ly produced in situ or externally supplied through quercetin (p<0.001). The leaf extract exhibited foods and/or supplements. Endogenous and ex- better H O scavenging and Fe2+ chelating activ- 2 2 ogenous antioxidants act as free radical scav- ity than the other parts. The extracts exhibited good antioxidant activity in linoleic acid peroxi- engers by preventing and repairing damage dation test but were not comparable to vitamin caused by ROS and, therefore, can enhance the C (p<0.001). Extracts showed weak reducing immune defense and lower the risk of cancer and power activity. The stem extract showed better degenerative diseases2,4. The use of traditional antihaemolytic activity than the flower and leaf. medicine is widespread and plants still present a The flower extract had higher phenolic con- large source of natural antioxidants that might tents. The extracts exhibited different levels of serve as leads for the development of novel antioxidant and antihaemolytic activities in all 1 tested models. drugs . Consequently, the need to identify alter- Conclusions: This study showed remarkable native natural and safe sources of food antioxi- antioxidant and antihemolytic activities in Ferula dants arose and the search for natural antioxi- foetida. Biological effects may be attributed to dants, especially of plant origin, has notably in- the presence of phenols and flavonoids in the creased in recent years5,6. The Ferula genus (Um- extract. It is very promising for further biochemi- belliferae) has been found to be a rich source of cal experiments. gum-resin7. This resin enjoys a reputation as a 8 Key Words: folklore medicine . Sedative, carminative, anti- spasmodic digestive, expectorant, laxative, anal- Antioxidant activity, Chelating activity, DPPH, Ferula foetida, Flavonoid, Nitric oxide scavenging. gesic, anthelminitic, antiseptic and diuretic properties have been reported from the Ferula genus9.

Corresponding Author: Mohammadali A. Ebrahimzadeh, Ph.D; e-mail: [email protected] 157 S.M. Nabavi, M.A. Ebrahimzadeh, S.F. Nabavi, B. Eslami, A.A. Dehpour

It is also believed to have aphrodisiac and in- Determination of Total Phenolic creasing sexual appetite10. This genus presents Compounds and Flavonoid Content interesting phytochemical features, such as the Total phenolic compound contents were deter- occurrence of sesquiterpenes and sesquiterpene mined by the Folin-Ciocalteau method19. The ex- coumarins8,11. Ferula foetida regel is a perennial tract samples (0.5 ml) were mixed with 2.5 ml of plant, which blooms once in its several years of 0.2 N Folin-Ciocalteau reagents for 5 min and life12,13. It is native to central Asia, Afghanistan 2.0 ml of 75 g l-1 sodium carbonate was then and Iran12. Previously polysulfide derivatives14 added. The absorbance of reaction was measured and sesquiterpene coumarins15 were reported at 760 nm after 2 h of incubation at room temper- from Ferula foetida. In vitro fungi toxicity of the ature. Results were expressed as gallic acid resin extract of Ferula foetida has been studied16. equivalents. Total flavonoids were estimated as We have recently reported good antioxidant ac- previously described20. Briefly, 0.5 ml solution of tivity from Ferula assafoetida9 and Ferula gum- each extracts in methanol were separately mixed mosa17,18. The aim of this study was to determine with 1.5 ml of methanol, 0.1 ml of 10% alu- the antioxidant and antihemolytic activities of the minum chloride, 0.1 ml of 1 M potassium ac- hydroalcoholic extract of Ferula foetida regel etate, and 2.8 ml of distilled water and left at leaf, flower or stems in order to understand the room temperature for 30 min. The absorbance of usefulness of this plant as a medicinal plant. the reaction mixture was measured at 415 nm with a double beam spectrophotometer (UV–Visible EZ201, Perkin Elmer, USA). Total flavonoid contents were calculated as quercetin Materials and Methods from a calibration curve.

Chemicals Trichloroacetic acid (TCA), 1,1-diphenyl-2- Antioxidant activity picryl hydroxyl (DPPH), potassium ferricyanide and hydrogen peroxide (H2O2) were purchased DPPH Radical-Scavenging Activity from Sigma Chemicals Co. (St Louis, MO, USA). The stable 1, 1-diphenyl-2-picryl hydroxyl Butylated hydroxyanisole (BHA), ascorbic acid, radical (DPPH) was used for determination of sulfanilamide, N-(1-naphthyl) ethylenediamine free radical scavenging activity of the extracts21 dihydrochloride, ethylenediaminetetraacetic acid Different concentrations of extracts were added, (EDTA) and ferric chloride were purchased from at an equal volume, to the methanol solution of Merck (Darmstadt, Germany). All other chemi- DPPH (100 µM). After 15 min at room tempera- cals were of analytical grade or purer. ture, the absorbance was recorded at 517 nm (UV–Visible EZ201, Perkin Elmer, USA). The Plant Materials and Preparation of experiment was repeated three times. Vitamin C, Freeze-Dried Extract BHA and quercetin were used as standard con- Ferula foetida regel was collected from trols. IC50 values denote the concentration of Gadouk area, central Elburz, Iran, in 2009. The sample, which is required to scavenge 50% of sample was identified by Dr. Bahman Eslami DPPH free radicals. (Assistant Professor of plant systematic, Islamic Azad University, Ghaemshahr Branch, Iran). Determination of Metal Chelating Activity Voucher specimens were deposited in the The ability of the Ferula foetida regel extracts Herbarium of Sari Faculty of Pharmacy (No GRF to chelate ferrous ions was estimated in our re- 32-34). A known amount of each sample (250 g) cently published paper22. Briefly, different con- was extracted at room temperature for 24 h by centrations of each extract were added to a solu- percolation with ethanol/water (600 ml, 70/30 tion of 2 mM FeCl2 (0.05 ml). The reaction was v/v). The extract was then separated from the initiated by the addition of 5 mM ferrozine (0.2 sample residue by filtration through Whatman ml) and the mixture was then shaken vigorously No. 1 filter paper. This procedure was repeated and left to stand at room temperature for 10 min. thrice. The resulting extract was concentrated The absorbance of the solutions was measured over a rotary vacuum until a crude extract was spectrophotometrically at 562 nm (UV–Visible obtained, which was then freeze-dried for com- EZ201, Perkin Elmer, USA). The percentage in- plete solvent removal. hibition of ferrozine-Fe2+ complex formation was

158 Antioxidant and antihaemolytic activities of Ferula foetida regel (Umbelliferae)

calculated as [(A0 -A1)/A0] × 100, where A0 was Elmer, USA). Increased absorbance of the reac- the absorbance of the control, and A1 of the mix- tion mixture indicated increased reducing power. ture containing the extract or the absorbance of a Vitamin C was used as positive control. standard solution. EDTA was used as a standard. Antioxidant Activity in A Hemoglobin- Assay of Nitric Oxide-Scavenging Activity Induced Linoleic Acid Peroxidation Test Sodium nitroprusside (10 mM), in phosphate- The antioxidant activity of extracts was deter- buffered saline (PBS), was mixed with different mined by a modified photometry assay25. Reac- concentrations of extracts dissolved in water and tion mixtures (200 ml) containing 10 ml of each incubated at room temperature for 150 min. The extract (10-400 mg), 1 mmol/L of linoleic acid same reaction mixture, without the extracts but emulsion, 40 mmol/L of phosphate buffer (pH with an equivalent amount of water, served as 6.5), and 0.0016% hemoglobin, were incubated control. After the incubation period, 0.5 ml of at 37ºC for 45 min. After the incubation, 2.5 ml

Griess reagent (1% sulfanilamide, 2% H3PO4 and of 0.6% HCl in ethanol was added to stop the 0.1% N-(1-naphthyl) ethylenediamine dihy- lipid peroxidation. The amount of peroxide value drochloride) was added. The absorbance of the was measured in triplicate, using the thiocyanate chromophore formed was read at 546 nm. method by reading the absorbance at 480 nm af- 19,23 Quercetin was used as positive control . ter coloring with 100 ml of 0.02 mol/l of FeCl2 and 50 ml of ammonium thiocyanate (30 g/100 Scavenging of Hydrogen Peroxide ml). Vitamin C was used as positive control. The ability of the extracts to scavenge hydrogen peroxide was determined according to our Antihemolytic Activity of Extracts 9 recently published paper . A solution of hydro- Against H2O2 Induced Hemolysis gen peroxide (40 mM) was prepared in phos- The inhibition of rat erythrocyte hemolysis by phate buffer (pH 7.4). The concentration of hy- the extracts was evaluated according to the proce- drogen peroxide was determined by absorption at dure described by Ebrahimzadeh et al26. The rat

230 nm using a spectrophotometer. All the ex- erythrocyte hemolysis was performed with H2O2 tracts (0.1-3.2 mg ml-1) in distilled water were as free radical initiator. To 100 µl of 5% (v/v) sus- added to a hydrogen peroxide solution (0.6 ml, pension of erythrocytes in phosphate buffered 40 mM). The absorbance of hydrogen peroxide saline (PBS), 50 µl of each extracts with different at 230 nm was determined after ten minutes concentrations (5-25 µg in PBS pH 7.4), which against a blank solution containing phosphate corresponds to 100-3200 µg of extract, was added. buffer without hydrogen peroxide. The percent- Then, 100 µl of 100 mM H2O2 (in PBS pH7.4) age of hydrogen peroxide scavenging by the ex- was added to it. The reaction mixtures were shak- tracts and standard compounds was calculated as en gently while being incubated at 37°C for 3 h. follows: % scavenged [H2O2] = [(Ao −A1)/Ao] × The reaction mixtures were diluted with 8 ml of 100, where Ao was the absorbance of the control PBS and centrifuged at 2000×g for 10 min. The and A1 was the absorbance in the presence of the absorbance of the resulting supernatants was mea- sample of extract and standard. sured at 540 nm by spectrophotometer to determine the hemolysis. Likewise, the erythrocytes

Reducing Power Determination were treated with 100 µM H2O2 and without in- The reducing power of extract was determined hibitors (plant extracts) to obtain a complete he- according to our recently published papers24. 2.5 molysis. The absorbance of the supernatants was ml of extract (25-800 mg ml-1) in water were measured at the same condition. The inhibitory ef- mixed with phosphate buffer (2.5 ml, 0.2M, pH fect of the extracts was compared with standard

6.6) and potassium ferricyanide [K3Fe(CN)6] (2.5 antioxidant vitamin C. To evaluate the hemolysis ml, 1%). The mixture was incubated at 50°C for induced by extracts, erythrocytes were preincubat- 20 min. A portion (2.5 ml) of trichloroacetic acid ed with 50 µl of extracts corresponding to 25 µg (10%) was added to the mixture to stop the reac- extracts for 1 h and the hemolysis was determined. tion, which was then centrifuged at 3000 rpm for Percentage of hemolysis was calculated by taking

10 min. The upper layer of solution (2.5 ml) was hemolysis caused by 100 µM H2O2 as 100%. The mixed with distilled water (2.5 ml) and FeCl3 IC50 values were calculated from the plots as the (0.5 ml, 0.1%), and the absorbance was mea- antioxidant concentration required for the inhibi- sured at 700 nm (UV–Visible EZ201, Perkin tion of 50% hemolysis.

159 S.M. Nabavi, M.A. Ebrahimzadeh, S.F. Nabavi, B. Eslami, A.A. Dehpour

Statistical Analysis Experimental results are expressed as means ± SD. All measurements were replicated three times. The data were analyzed by an analysis of Flower variance (p<0.05) and the means separated by Vitamin C Duncan’s multiple range tests. The EC values Stem 50 Leaf were calculated from linear regression analysis.

Results Abssorbance at 700 nm

Total phenol and flavonoids contents of extracts obtained from Ferula foetida leaf, flower, and stem are shown in Table I. The maximum of Concentration (µg ml-1) extractable polyphenol content was recorded in flower with 51.7 ± 2.3 mg gallic acid equiva- Figure 1. Reducing power of flower, stem and leaf of Fer- lent/g of extract, by reference to standard curve ula foetida regel. Vitamin C has been used as control. (y = 0.0063x, r2 = 0.987). In addition, maximum flavonoid contents were recorded in leaf extract with 20.9 ± 1.4 mg quercetin equivalent/g of ex- not show any harmful effects on erythrocytes. tract, by reference to standard curve (y = 0.0067x Results are shown in Table I. Stem extract 2 + 0.0132, r = 0.999). IC50 for DPPH radical- showed better antihemolytic activity than other -1 scavenging activity exists in Table 1. The IC50 extracs (IC50 was 248.7± 16.2 µg ml vs. vitamin values for vitamin C, quercetin and BHA were C which was 235 ± 9 µg ml-1). 5.05 ± 0.1, 5.28 ± 0.2 and 53.96 ± 3.1mg ml-1, respectively. Results of iron chelating capacity were presented in Table I. Among the extracts, leaf extract showed better activity than others Discussion -1 with IC50 = 302.2 ± 13.6 µgml . EDTA showed -1 very powerful activity (IC50 = 18 ± 1.5 µgml ). In Extracts showed high level of total phenol and scavengers of nitric oxide, percentage of inhibi- flavonoid contents. Phenols and polyphenolic tion was increased with increasing concentration compounds, such as flavonoids, are widely found of the extracts but the activity was very weak -1 (IC50 for stem extract was 896.9 ± 21.9 µgml vs. quercetin which was 20 ± 0.1 µgml-1). The extracts were capable of scavenging hydrogen peroxide in a concentration dependent manner. Re- sults exist in Table 1. Leaf extract showed better Flower activity than others (IC was 105.7 ± 4.7 mg ml- Vitamin C 50 Stem 1 ). The IC50 values for vitamin C and quercetin Leaf were 21.4 ± 1.1 and 52 ± 2.6 mg ml-1, respectively. Figure 1 shows the dose response curves for the reducing powers of the extracts. It was found that the reducing powers of all extracts increased with the increase of their concentrations. There were no significant differences (p>0.05) among Inhibitory ability (%) the extracts in reducing power. Their activities were not comparable with vitamin C (p<0.001). Tested extracts showed good activity in hemoglo- Concentration (μg ml-1) bin-induced linoleic acid system but there were significant differences between extracts and vita- Figure 2. Effect of flower, stem and leaf extracts of Ferula min C (p<0.01) (Figure 2). The effects of ex- foetida regel on hemoglobin-catalyzed linoleic acid peroxi- tracts were tested and it was found that they did dation test. Vitamin C has been used as control.

160 Antioxidant and antihaemolytic activities of Ferula foetida regel (Umbelliferae) ) Sample -1 ) g (mg -1 (mg g for vitamin C and quercetin were 21.4 ± 1.1 52 50 IC c a ) -1 Ferula foetida regel. Ferula (µg ml 50 IC of quercetin was 20 ± 0.1. of quercetin was 50 IC b b ) -1 (mg ml 50 IC of vitamin C was 235 ± 9. of vitamin C was 50 IC f c ). ) -1 -1 (µg ml scavenging Nitric oxide radical DPPH free flavonoid Total phenol Total activity, scavenging, scavenging, contents contents 2 50 O IC 2 = 18 ± 1.5 µg ml 50 50 d ) -1 chelating H 2+ EDTA used as control (IC EDTA d ) ml (µg -1 Phenol and flavonoid contents, antioxidant and antihemolytic activities of flowers, stems and leaves of stems and leaves of flowers, contents, antioxidant and antihemolytic activities Phenol and flavonoid activity ability, IC activity ability, (µg ml of BHA was 53.96 ± 3.1, vitamin C, 5.05 ± 0.1 and quercetin 5.28 ± 0.2, respectively. 53.96 ± 3.1, vitamin C, 5.05 0.1 and quercetin 5.28 0.2, respectively. of BHA was Antihemolytic Fe 50 260.4 ± 11.5253 ± 9.4248.7 ± 16.2 775.5 ± 21.4 302.2 ± 13.6 619.6 ± 18.9 193.8 ± 8.6 105.7 ± 4.7 150.2 ± 5.2 1.2 ± 0.04 0.89 ± 0.01 1.4 ± 0.03 471.6 ± 19.4 775.9 ± 26.3 192.5 ± 8.6 20.7 ± 0.9 9.9 ± 0.3 20.9 ± 1.4 51.7 ± 2.3 36.7 ± 1.3 49.4 ± 2.1 Flower Stem Leaf IC Table I. Table ± 2.6, respectively. ± 2.6, respectively. a

161 S.M. Nabavi, M.A. Ebrahimzadeh, S.F. Nabavi, B. Eslami, A.A. Dehpour in food products, derived from plant sources, and presence of antioxidants in the samples would re- they have been shown to possess significant an- sult in the reducing of Fe3+ to Fe2+ by donating an tioxidant activities9. DPPH is a stable nitrogen- electron. Amount of Fe2+ complex can be then be centered free radical, the color of which changes monitored by measuring the formation of Perl’s from violet to yellow upon reduction by either Prussian blue9 at 700 nm. Increasing absorbance the process of hydrogen- or electron- donation. indicates an increase in reductive ability. Reduc- Substances which are able to perform this reac- ing powers of extracts increased with the in- tion can be considered as antioxidants and there- crease of their concentrations, but their activities fore radical scavengers27,28. Phenol and flavonoid were not comparable with vitamin C (p<0.001). contents of this plant may lead to its agreable Polyphenolic contents of all the sample extracts DPPH-scavenging activity26. Iron chelators re- appear to function as good electron and hydrogen duce iron-related complications in human and atom donors and, therefore, should be able to ter- thereby improves quality of life and overall sur- minate radical chain reaction by converting free vival in some diseases such as thalassemia major radicals and reactive oxygen species to more sta- or Alzheimer’s disease29. Bivalent transition met- ble products. Similar observation between the al ions play an important role as catalysts of ox- polyphenolic constituents in terms of dose de- idative processes30. These processes can be dependent and reducing power activity have been layed by iron chelation and deactivation. The reported for several plant extracts9. transition metal, iron, is capable of generating Erythrocytes are considered as prime targets free radicals from peroxides by Fenton reactions for free radical attack, owing to the presence of and may be implicated in human cardiovascular both high membrane concentration of polyunsatu- 31 2+ diseases . Thus, minimizing Fe concentration rated fatty acids (PUFA) and the O2 transport as- affords protection against oxidative damage. sociated with redox active hemoglobin molecules,

Many researches focused on some natural prod- which are potent promoters of reactive O2 species. ucts, especially flavonoids that possess direct in- Specially linoleic acid and arachidonic acid are fluence on iron (III) ions level within tissues29. targets of lipid peroxidation35. The inhibition of Ferrozine can quantitatively form complexes lipid peroxidation by antioxidants may be due to with Fe2+. In the presence of other chelating their free radical-scavenging activities. Superox- agents, the complex formation is disrupted with ide indirectly initiates lipid peroxidation because the result that the red color of the complexes de- superoxide anion acts as a precursor of singlet creases. In this assay, both extract and EDTA in- oxygen and hydroxyl radical36,37. Hydroxyl radi- terfered with the formation of ferrous and fer- cals eliminate hydrogen atoms from the mem- rozine complex, suggesting that it has chelating brane lipids, which results in lipid peroxidation. activity and captures ferrous ion before ferrozine. Tested extracts show good activity in hemoglo- The nitric oxide assay is based on the principle bin-induced linoleic acid peroxidation test. Effect that sodium nitroprusside in aqueous solution at of extracts were tested and found that they did not physiological pH spontaneously generates nitric show any harmful effects on erythrocytes. oxide which interacts with oxygen to produce ni- Flavonoids interactions with cell membranes trite ions that can be estimated using Griess which generally serve as targets for lipid peroxi- reagent. Scavengers of NO compete with oxy- dation (LP), constitute an important area of re- gen, leading to reduced production of nitrite ions. search38. Various model membrane systems like Extracts did not show any in scavenging NO. LDL and red blood cells (RBC) membrane com-

Scavenging of H2O2 by extract may be attrib- prising physiologically important membrane uted to its phenolics, and other active compo- protein components offer a physiologically rele- nents which can donate electrons to H2O2, thus vant and a relatively simple system for studying 32,33 39 neutralizing it to water . Although H2O2 itself LP . RBC has been chosen as an in vitro model is not very reactive, it can sometimes cause cyto- to study the oxidant/antioxidant interaction since toxicity by giving rise to hydroxyl radicals in the its membrane is rich in polyunsaturated fatty cell. Thus, removing H2O2 is very important acids, which are extremely susceptible to peroxi- throughout food systems9. Membrane lipids are dation40. During recent years, a few interesting rich in unsaturated fatty acids that are most sus- studies have been reported, indicating the protec- ceptible to oxidative processes. Specially, linole- tive effects of some plants extracts against oxida- ic acid and arachidonic acid are targets of lipid tive damage in intact RBC membranes26,37,40,41. peroxidation34. In the reducing power assay, the Hemolysis has a long history of use in measuring

162 Antioxidant and antihaemolytic activities of Ferula foetida regel (Umbelliferae) free radical damage and its inhibition by antioxi- 7) FERNCH D. ETHNOBOTHANY OF THE UMBELLIFERAE. IN: dants but only few studies have been performed HEYWOOD VH, ED., The Chemistry and Biology of the Umberifella. Academic press, London. 1971: with erythrocytes in whole blood. In this study, pp 285-412. we used a biological test based on free radical-induced erythrocytes lysis in rat blood. Lipid oxi- 8) ABD EL-RAZEK MH, OHTA S, AHMED AA, HIRATA T. Sesquiterpene coumarins from the roots of Ferula dation of rat blood erythrocyte membrane medi- assafoetida. Phytochemistry 2001; 58: 1289- ated by hydrogen peroxide induces membrane 1295. damage and subsequently hemolysis. Stem ex- 9) DEHPOUR AA, EBRAHIMZADEH MA, NABAVI SF, NABAVI tract showed better antihemolytic activity than SM. Antioxidant activity of the methanol extract of -1 other extracts (IC50 was 248.7± 16.2 µg ml vs. Ferula assafoetida and its essential oil composi- vitamin C which was 235 ± 9 µg ml-1). The anti- tion. Grasas Aceites 2009; 60: 405-412. hemolytic activity of flavonoids has been previ- 10) EIGNER D, SCHOLZ D. Das Zauberbchlein der Gyani ously reported and activity of the extract maybe Dolma. Pharmazie in userer Zeit 1990; 19: 141-152. 9,38 results in high flavonoid content . 11) SU BN, TAKAISHI Y, H ONDA G, ITOH M, TAKEDA Y, KODZHIMATOV OK, ASHURMETOV O. Sesquiterpene phenylpropanoid and sesquiterpene chromone derivatives from Ferula pallida. J Nat Prod 2000; Conclusions 63: 520-522. 12) MOZAFFARIAN V. editor. In: A Dictionary of Iranian Our study improved remarkable antioxidant Plant Names. Tehran. Farhang Moaser. 2006, p. and antihemolytic activities in hydroalcoholic ex- 228. tract of Ferula foetida regel Boiss flower, stem 13) ZARGARI A. Medicinal Plants. Vol. 2. Tehran: Tehran and leaf. These effects maybe result of their high University Publication. 1991. pp. 598-602. phenol and flavonoids contents. It is therefore 14) DUAN H, TAKAISHI Y, T ORI M, TAKAOKA S, HONDA G, very promising for further biochemical experi- ITO M, TAKEDA Y, K ODZHIMATOV OK, KODZHIMATOV K, ASHURMETOV O. Polysulfide derivatives from Ferula ments, which will be focused on evaluating the foetida. J Nat Prod 2002; 65: 1667-1669. mechanism of this activity. 15) ABD EL-RAZEK MH, WU YC, CHANG FR. Sesquiter- pene Coumarins from Ferula foetida. J Chin Chem Soc 2007; 54: 235-238.

16) RAM D. Fungitoxicity of some plants extract against Alternaria brassicae. Ann Agric Biol Res References 1997; 2: 25-26.

17) NABAVI SF, EBRAHIMZADEH MA, NABAVI SM, ESLAMI B. 1) HUANG DJ, LIN CD, CHEN HJ, LIN YH. Antioxidant Antioxidant activity of flower, stem and leaf extract and antiproliferative activities of sweet potato of Ferula gummosa Boiss. Grasas Aceites 2010; (Ipomoea batatas [L.] Lam “Tainong 57”) con- 61: 244-250. stituents. Bot Bull Acad Sin 2004; 45: 179-186. 18) EBRAHIMZADEH MA, NABAVI SM, NABAVI SF, DEHPOUR 2) PHAM-HUY LA, HE H, PHAM-HUYC C. Free radicals, AA. Antioxidant activity of hydroalcholic extract of antioxidants in disease and health. Int J Biomed Ferula gummosa Boiss roots. Eur Rev Med Phar- Sci 2008; 4: 89-96. macol Sci 2011; in press.

3) TEPE B, SOKMEN A. Screening of the antioxidative 19) NABAVI SM, EBRAHIMZADEH MA, NABAVI SF, FAZELIAN properties and total phenolic contents of three en- M, ESLAMI B. In vitro antioxidant and free radical demic Tanacetum subspecies from Turkish flora. scavenging activity of Diospyros lotus and Pyrus Bioresource Technol 2007; 98: 3076-3079. boissieriana growing in Iran. Pharmacog Mag 2009; 4: 123-127. 4) WILLCOX JK, ASH SL, CATIGNANI GL. Antioxidants and prevention of chronic disease. Crit Rev Food Sci 20) EBRAHIMZADEH MA, MAHMOUDI M, AHANGAR N, EHTE- Nutr 2004; 44: 275-295. SHAMI S, ANSAROUDI F, N ABAVI SF, NABAVI SM. Antide- pressant activity of corn silk. Pharmacologyonline 5) BRAHIMZADEH ABAVI ABAVI SLAMI E MA, N SM, N SF, E SH, 2009; 3: 647-652. BEKHRADNIA AR. Mineral elements and antioxidant activity of three locally edible and medicinal 21) EBRAHIMZADEH MA, NABAVI SM, NABAVI SF, ESLAMI B. plants in Iran. Asian J Chem 2010; 8: 6257-6266. Antioxidant activity of O. Sintenisii bulbs and aeri- al part at flowering stage. Trop J Pharmac Res 6) HASEMI HASEMI BRAHIMZADEH Antioxi- G K, G Y, E MA. 2010; 9: 141-148. dant activity, phenol and flavonoid contents of 13 Citrus species peels and tissues. Pak J Pharm 22) EBRAHIMZADEH MA, NABAVI SM, NABAVI SF. Correla- Sci 2009; 22: 277-281. tion between the in vitro iron chelating activity

163 S.M. Nabavi, M.A. Ebrahimzadeh, S.F. Nabavi, B. Eslami, A.A. Dehpour

and poly phenol and flavonoid contents of some 32) HALLIWELL B, GUTTERIDGE JMC. Role of free radicals medicinal plants. Pak J Biol Sci 2009; 12: 934- and catalytic metal ions in human disease: an 938. overview. Method Enzymol 1990; 186: 1-85.

23) EBRAHIMZADEH MA, NABAVI SF, NABAVI SM. Antioxi- 33) EBRAHIMZADEH MA, NABAVI SM, NABAVI SF, ESLAMI B, dant activities of methanol extract of sambucus EHSANIFAR S. Antioxidant activity of Hyoscyamus ebulus L. flower. Pak J Biol Sci 2009; 12: 447- squarrosus fruits. Pharmacologyonline 2009; 2: 450. 644-650.

24) EBRAHIMZADEH MA, NABAVI SF, NABAVI SM, ESLAMI B. An- 34) EBRAHIMZADEH MA, NABAVI SM, NABAVI, SF, ESLAMI B. tihypoxic and antioxidant activity of Hibiscus escu- Antioxidant activity of aqueous extract of Pyrus lentus seeds. Grasas Aceites 2010; 61; 30-36. boissieriana fruit. Pharmacologyonline 2009; 1: 1318-1323. 25) YUAN X, WANG J, YAO H, CHEN F. Free radical-scavenging capacity and inhibitory activity on rat ery- 35) YU LL. Free radical scavenging properties of con- throcyte hemolysis of feruloyl oligosaccharides jugated linoleic acids. J Agr Food Chem 2001; 49: from wheat bran insoluble dietary fiber. LWT-Food 3452-3456. Sci Technol 2005; 38: 877-883. 36) EBRAHIMZADEH MA, EHSANIFAR S, ESLAMI B. Sambucus 26) EBRAHIMZADEH MA, NABAVI SF, NABAVI SM. Antihe- ebulus elburensis fruits: A good source for antioxi- molytic and antioxidant activity of Hibiscus escu- dants. Pharmacog Mag 2009; 4: 213-218. lentus leaves. Pharmacologyonline 2009; 2: 1097- 37) EBRAHIMZADEH MA, NABAVI SF, NABAVI SM, ESLAMI 1105. B. Antihemolytic and antioxidant activities of Al- 27) EBRAHIMZADEH MA, NABAVI SM, NABAVI SF, ESLAMI B. lium paradoxum. Cent Eur J Biol 2010; 5: 338- Free radical scavenging ability of methanolic ex- 345. tract of Hyoscyamus squarrosus leaves. Pharma- 38) CHAUDHURI S, BANERJEE A, BASU K, SENGUPTA B, SEN- cologyonline 2009; 2: 796-802. GUPTA PK. Interaction of flavonoids with red blood 28) NABAVI SM, EBRAHIMZADEH MA, NABAVI SF, JAFARI M. cell membrane lipids and proteins: antioxidant Free radical scavenging activity and antioxidant and antihemolytic effects. Int J Biol Macromol capacity of Eryngium caucasicum Trautv and 2007; 41: 42-48. Froriepia subpinnata. Pharmacologyonline 2008; 39) SHIVA SHANKAR REDDY CS, SUBRAMANYAM MV, VANI R, 3: 19-25. ASHA DEVI S. In vitro models of oxidative stress in 29) GRAZUL M, BUDZISZ E. Biological activity of metal rat erythrocytes: effect of antioxidant supple- ions complexes of chromones, coumarins and ments. Toxicol In Vitro 2007; 21: 1355-1364. flavones. Coordin Chem Rev 2009; 253: 2588- 40) EBRAHIMZADEH MA, RAHMANI Z, ESLAMI B, NABAVI SF, 2598. NABAVI SM. Antioxidant and antihemolytic activities 30) HALLIWELL B. Antioxidants: the basics- what they of Crusianella sintenisii. Pharmacologyonline are and how to evaluate them. Adv Pharmacol 2009; 3: 716-723. 1997; 38: 3-20. 41) EBRAHIMZADEH MA, NABAVI SF, ESLAMI B, NABAVI SM. 31) EBRAHIMZADEH MA, NABAVI SF, NABAVI SM. Essential Antioxidant and antihemolytic potentials of oil composition and antioxidant activity of Ptero- Physosperum cornubiense (L.) DC. Pharmacolo- carya fraxinifolia. Pak J Biol Sci 2009; 12: 957-963. gyonline 2009; 3: 394-403.

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