J Arthropod-Borne Dis, June 2018, 12(2): 119–126 F Yousefbeyk et al.: Antioxidant and … 119 http://jad.tums.ac.ir Published Online: June 12, 2018 Original Article Antioxidant and Larvicidal Activity of Areal Parts of Scrophularia striata against Malaria Vector Anopheles stephensi Fatemeh Yousefbeyk 1, Hassan Vatandoost 2, Fereshteh Golfakhrabadi 3, Zahra Mirzaee 3, Mohammad Reza Abai 2, Gholamreza Amin 3, *Mahnaz Khanavi 3, 4 1Department of Pharmacognosy, School of Pharmacy, Guilan University of Medical Sciences, Rasht, Iran 2Department of Medical Entomology and Vector Control, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran 3Department of Pharmacognosy, Faculty of Pharmacy and Persian Medicine and Pharmacy Research Center, Tehran University of Medical Sciences, Tehran, Iran 4Faculty of Land and Food Systems University of British Columbia, British Columbia, Canada (Received 13 May 2015; accepted 30 Apr 2018) Abstract Background: Scrophularia striata is a perennial plant which is native in all parts of Iran, Turkey, and Azerbaijan. In this study, the total phenol content, antioxidant and larvicidal activities of total extract and different fractions of this plant were evaluated. Methods: The aerial parts of S. striata were collected from Boli village, Illam Province, western Iran in Apr 2013. The total phenol content of total extract and different fractions were evaluated by Folin-Ciocalteu method. Moreover, antioxidant activity was tested by DPPH and FRAPS assays. Larvicidal activity was investigated according to stand- ard method described by WHO. Results: Ethyl acetate fraction (EF) had the highest content of total phenol (75.9±0.06mg Gallic acid equivalent/g dry extract). Furthermore, among the tested extract, methanol-water fraction (MWF), total methanol extract (TME) and water fraction (WF) showed the highest antioxidant activity in the DPPH assay (IC50= 226.8, 283.66 and 299.4 μg.ml-1, respectively). In FRAP assay MWF and WF and TME had the highest antioxidant activities (664.4±0.002, 565.3±0.003, 519.5±0.003mmol FeII/g dry extract, respectively). Ethyl acetate fraction had maximum larvicidal activity (LC50 49.1ppm) followed by TME (LC50 64.26ppm) and hexane fraction (HF) (LC50 89.69). Conclusion: Scrophularia striata collected from west of Iran illustrated considerable antioxidant and larvicidal ef- fects and further in vitro and in vivo experimental models for investigation would be required. Keywords: Scrophularia striata, Antioxidant activity, Larvicidal activity, Anopheles stephensi Introduction According to the word malaria report, in 2014, 198 million cases of malaria occurred globally in 2013 and led to 584000 deaths (1-3). In southern parts of Iran including Sistan and Baluchistan, Hormozgan and some parts of Kerman provinces, this disease remains as a vital public health problem (4). Six Anoph- eles vectors exist in this area including An. culicifacies, An. stephensi, An. dthali, An. flu- viatilis, An. superpictus and An. pulcherrimus (5). The present proliferation of malaria is due basically to increasing resistance of mosqui- toes to current insecticides (6). Plant materials have been investigated for their susceptibility in controlling the malaria vector due to their ovicidal, larvicidal and adulticidal activities. The most susceptible stage to attack mosquitoes is larval stage as they are concentrated in smaller areas. There- fore, interrupting mosquito life cycle at lar- val stage is one of the important methods for controlling malaria transmission (7). *Corresponding author: Dr Mahnaz Khanavi, E- mail: khanavi@mail.ubc.ca/ khanavim@tums.ac.ir http://jad.tums.ac.ir/ J Arthropod-Borne Dis, June 2018, 12(2): 119–126 F Yousefbeyk et al.: Antioxidant and … 120 http://jad.tums.ac.ir Published Online: June 12, 2018 The genus Scrophularia (Scrophulariaceae) comprises about 300 known species. The var- ious species of Scrophularia genus have been traditionally used in medical conditions in- cluding skin inflammatory disease like sca- bies, tumors, eczema (8), and psoriasis, inflam- matory disorders fever, constipation, pharyn- gitis, neuritis, laryngitis affections (9). Among these species, S. ningpoensis Hemsl has been used for treatment of fever laryngitis, phar- yngitis, neuritis and constipation swelling in China. Moreover, S. grossheimi and S. no- dosa are used as diuretic agents (10). Recent studies reported antimalarial, antiprotozoal and antimycobacterial activities of S. cryptophila (11). Phytochemical studies indicated the pres- ence of saikosaponins, iridoids and phenylpro- panoid glycosides such as angoroside A, an- goroside C, angoroside D, acteoside and iso- acteoside in S. scorodonia L. (8), iridoid glu- cosides including catalpol, 6-O-methylcatal- pol and aucubin, phenylethanoid glycoside like angoroside C in S. lepidota (12), resin glycosides including crypthophilic acids A– C in S. cryptophila (11). Many natural compounds have been re- ported for larvicidal activity. For example, flavonoids namely poncirin, rhoifolin, nar- ingin and marmesin from Poncirus trifoliate (13), lupinfolin and rotenone and deguelin from Derris trifoliate (14), new iridoid gly- cosides like 6'-O-rhamnosylharpagide and 6- O-xylosylharpagoside-B form Ajuga remota (15) and coumarins such as umbelliferone, herniarin, psoralen and xanthotoxin from Cnid- ium monnieri (16) and coumarin derivative pachyrrhizine from Neorautanenia mitis (17). Scrophularia striata Boiss is a perennial plant grown in all parts of Iran as well as Turkey and Azerbaijan. These species have been used to cure different inflammatory dis- eases such as allergy, rheumatics and chronic inflammatory disorders in Iranian folk med- icine (18). So far, phytochemical investiga- tion revealed the presence of cinnamic acid, flavonoids such as quercetine, isorhamnetin- 3-O-rutinoside, and nepitrinandand, and one phenyl propanoid glycoside acteoside 1 in this plant (10). Studied showed the inhibitory ef- fect of S. striata extract on matrix metallo- proteinases and astrocyte cancer cell line (1321) (18, 19). Different extracts from the aerial part of S. canina were evaluated for the insecticidal activity against larvae and adult females of Culex pipiens molestus. The most toxicity was revealed by the petroleum ether extract against second-instar larvae (48h, LC50= 23.5 ppm) and by the hexane extract against fourth- instar larvae (48h, LC50= 23.6ppm) (20). The leaves of S. nodosa showed larvicidal activ- ity against Cx. quinquefasciatus (75.3% mor- tality) (21). The ethanol root extract of S. lep- idota exhibited anti-protozoal activity (IC50 40.6μg.ml-1) and plasmodial enoyl-ACP re- ductase (FabI) enzyme inhibitory activity (IC50 100μg.ml-1), a key enzyme of fatty acid bio- synthesis in Plasmodium falciparum (12). In the present study, total extract and dif- ferent fractions of S. striata were investigat- ed for larvicidal properties against main ma- laria vector, An. stephensi. In addition, total phenol content and antioxidant activity (using DPPH and FRAP methods) of this plant were evaluated. Materials and Methods Plant material and extraction The aerial parts of S. striata were collected from Boli village, Ilam Province, western Iran in Apr 2013. The voucher specimen was de- posited in herbarium of faculty of Pharmacy, Tehran University of Medical Sciences (Her- barium number: 6748-TEH). The aerial parts (500g) were aired dried in shade and fractionated with hexane, ethyl acetate, methanol, methanol-water (1:1) and water at room temperature, consequently. To- tal methanol extract was prepared from aerial http://jad.tums.ac.ir/ J Arthropod-Borne Dis, June 2018, 12(2): 119–126 F Yousefbeyk et al.: Antioxidant and … 121 http://jad.tums.ac.ir Published Online: June 12, 2018 parts by percolation method. All the extract and fractions were dried using rotary evap- orator to give 2.8g, 6.5g, 60g, 48.2g, 7.7g and 28.7g residues from hexane, ethyl ace- tate, methanol, methanol-water (1: 1), water fractions and total extract, respectively. Antioxidant activity DPPH radical-scavenging activity assay The antioxidant activity of extracts was in- vestigated by the DPPH (2, 2′-diphenyl-1-pic- rylhydrazyl) free radical scavenging method ac- cording to an established protocol. Sample so- lutions (1ml) were prepared in methanol at dif- ferent concentration. Then the solutions added to DPPH methanol solution (2ml, 40μg.ml-1) and incubated at room temperature for 30 min. The absorbance was measured at 517nm. Vitamin E and butyl hydroxyanisole (BHA) were used as positive controls. IC50 values (indicate the concentration of the test sam- ples providing 50% radical scavenging) were calculated from graph-plotted scavenging per- centage against extract concentration (22). Ferric reducing antioxidant potential (FRAP scavenging) assay The FRAP assay was done by using the method described (23, 24). The FRAP rea- gent contained 5 ml of a (10mmol.l-1) TPTZ (2, 4, 6-tripyridyl- s- triazine) solution in 40 mmol.l-1 HCl plus 5ml of (20mmol.l-1) FeCl3 and 50ml of (0.3 mmol.l-1) acetate buffer, pH 3.6 was prepared freshly. Aliquots of each extract (50μl) were mixed with FRAP reagent (1.5ml). The mixtures were incubated at 37 °C, for 10min. Finally, the absorbance of each sample was measured at 593nm. For construc- tion of calibration curve, five concentrations of FeSO4, 7H2O (125, 250, 500, 750, 1000 mmol.l-1) were prepared to used. The antiox- idant activities were calculated as the con- centration of antioxidants having a reducing ability equivalent for 1mmol.l-1 FeSO4 (22, 25). Determination of total phenolic contents Total phenolic contents of extracts were determined by Folin-Ciocalteu method (26). Folin-Ciocalteu (Merk) reagent diluted to ten- fold with distilled water. Then 5ml of this so- lution was added to 1ml of each extract (1 mg.ml-1) and allowed to stand at room tem- perature for 10min. A 4ml sodium bicarbonate solution (75g.l-1) was added to the mixture. After 30min at room temperature, absorbance was measured at 765nm using a UV spectro- photometer (Pharmacia Biotech). Total phe- nolic contents were quantified by calibration curve obtained by measuring the absorbance of a known concentration of Gallic acid (GA) standard (20–200mg.l-1). The concentra- tions are expressed as milligrams of Gallic acid equivalents (GA) per gr dry extract (22, 27). Bioassays and larval mortality Fourth instar larvae of An. stephensi Ban- dar-Abbas strain was exposed to test concen- trations of 20, 40, 80, 160 and 320ppm of each extract (solvent: Ethanol) for 24h according to standard method described by WHO (1981). Briefly, 1ml of appropriate dilution of each extract with 224ml of water and 25 larvae in 25ml water mixed and total volume was 250 ml (2). For control, only 1ml of ethanol with 224ml of water and 25 larvae in 25ml water mixed and total volume was 250ml. The ex- periment was repeated four times on differ- ent days. The percentage of mortality was re- ported from the average for the four replicates after 24h exposure period. From the regres- sion line between logarithmic dose and probit mortality, the LC50 was measured (28, 29). The investigation of larvicidal activity has been carried out in the insectarium of De- partment of Medical Entomology and Vector Control, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran. http://jad.tums.ac.ir/ J Arthropod-Borne Dis, June 2018, 12(2): 119–126 F Yousefbeyk et al.: Antioxidant and … 122 http://jad.tums.ac.ir Published Online: June 12, 2018 Results Ethyl acetate fraction (EF) had the highest content of total phenol (75.9±0.06mg Gallic acid equivalent/g dry extract) (Table 1). Among the tested extract, methanol-water fraction (MWF), total methanol extract (TME) and wa- ter fraction (WF) showed highest antioxidant activity in the DPPH assay (IC50= 226.8, 283.66 and 299.4μg.ml-1, respectively). In FRAP assay MWF and WF and TME had the highest anti oxidant activities (664.4±0.002, 565.3±0.003, 519.5±0.003mmol FeII/g dry extract, respec- tively). The total extract and fractions of S. striata were effective against An. stephensi with LC50 ranged between 49.15 to 1265.96ppm. Ethyl acetate fraction had maximum larvicidal ac- tivity (LC50 49.15ppm) followed by TME (LC50 64.2623ppm) and HF (LC50 89.69) (Table 2). Table 1. Antioxidant activity and total phenolic content of total methanol extract and different fractions from aerial parts of Scrophularia striata DPPH (IC50: μg.ml-1) FRAP (mmol FeII/g dry extract) Total phenol contents (mg GAE/g dry extract) TME 283.7 519.5±0.003 64.8±0.03 HF >1000 248.8±0.035 -a EF 505.2 444.28±002 75.9±0.06 MF 511.3 513.7±0.026 41.7±0.04 MWF 226.8 664.4±0.002 56.2±0.02 WF 299.4 565.3±0.003 33±0.06 Key to extracts employed: TME: Total methanol extract, HF: hexane fraction, EF: ethyl acetate fraction, MF: methanol fraction, MWF: methanol-water fraction, WF: water fraction. a not detected. Table 2. Probit regression line parameters of total extract and different fractions of Scrophularia striata against Anopheles stephensi Intercept Slope ± SE LC50 (ppm) , 95%C.I. LC90 (ppm) , 95%C.I. λ2 table (df) P-value TME -4.5092 2.4941±0.537 64.2623 (27.2325-131.1766) 209.8017 (109.8552-2827.5650) 24.405* 0.001 HF -2.4518 1.2556±0.144 89.6911 (72.3973-112.2429) 940.7311 (577.0739-1990.1654) 13.345* 0.001 EF -2.7150 1.6051±0.156 49.1497 (40.1770-58.6299) 309.0209 (231.3864-462.1751) 13.345* 0.001 MF -4.6954 1.9658±0.199 244.6640 (203.2745-310.4233) 1097.8066 (747.3601-1921.5333) 13.345 0.001 MWF -6.5143 2.0977±0.424 1265.9601 (607.7026-2645.3743) 5161.5548 (2523.4135-75009.0794) 13.345 0.001 WF -7.9868 2.6359±0.195 1071.3972 (949.6278-1205.7662) 3282.1074 (2756.2862-4091.2804) 13.345 0.001 Key to extracts employed: TME: Total methanol extract, HF: hexane fraction, EF: ethyl acetate fraction, MF: methanol fraction, MWF: methanol water fraction, WF: water fraction, LC50 ±95% C.L= lethal dose cause 50% mortality, 95% confidence interval, LC90 ±95% C.I= lethal dose cause 90% mortality, 95% confidence interval, (df)= degree of freedom, P= P-value http://jad.tums.ac.ir/ J Arthropod-Borne Dis, June 2018, 12(2): 119–126 F Yousefbeyk et al.: Antioxidant and … 123 http://jad.tums.ac.ir Published Online: June 12, 2018 Discussion Scrophularia striata is a perennial plant which is native in all parts of Iran as well as Turkey and Azerbaijan. In this study total methanol extract and different fractions from aerial parts of S. striata were investigated for antioxidant and larvicidal activities. Accord- ing to our results, EF and TME had the highest amount of total phenolic compounds. Scroph- ularia steriata, as highest amount of total phenolic compounds, was reported in etha- nol 70% and ethyl acetate extracts (79.7 and 65.5mg Gallic acid equivalent/g dry extract, respectively). Moreover, water extract of this plant was reported to have better radical scav- enging activity (IC50 195µg.ml -1) (30). Furthermore, evaluation of larvicidal ac- tivity of S. striata revealed that LC50 value of total extract and fractions ranged between 49.15 to 1265.96ppm, and the EF had maxi- mum larvicidal activity (LC50 49.15ppm). Es- sential oils and extracts of different plant spe- cies have been investigated for larvicidal ac- tivity. For instance, the essential oil prepared from seeds of Heracleum persicum had lar- vicidal effect with LC50 value of 104.80ppm. The essential oil of Cupressus arizonica re- vealed significant larvicidal activity against An. stephensi with LC50 79.30ppm (31). More- over, for Coriandrum sativum LC50 value of 120.95ppm and for Cymbopogon olivieri 321.90ppm were reported (5), which their ac- tivities were low when compared with EF and TME of S. striata. The LC50 of essential oils from Tagetes minuta and Foeniculum vulgare were 1.05 and 20.10ppm, respectively (5), these plants were more effective than S. striata. The methanol extract of T. minuta had bet- ter LC50 value (2.5ppm) followed by metha- nol and aqueous extract of Nelumbo nucifera (8.89 and 11.82ppm, respectively) and metha- nol extract of Cassia fistula (17.97ppm) when compared with the extract and fractions of S. striata (32-34). Different extracts of fruits and leaves of Centratherum anthelminticum were tested for larvicidal activity against An. stephensi and the petroleum ether extract of fruits (LC50 162.60) were more toxic than that of leaf ex- tract (LC50 522.94) (7). Petroleum ether ex- tract from leaves of Artemisia annua had larvicidal activity with LC50 value of 263 ppm (7). The leaf and seed methanol extracts of Clitoria ternatea showed dose-dependent larvicidal activity against An. stephensi with LC50 values of 555.6 and 116.8ppm, respec- tively (35). The leaves petroleum ether ex- tract of Gymnema sylvestre exhibited the high- est mortality in the concentration of 1000ppm against the larvae of An. subpictus (LC50 166.28 ppm) (36). Moreover, the leave methanol ex- tract of Calotropis gigantea showed larvo- cidal activity (LC50 121.69ppm) (37). Larvi- cidal activity of aceton extract of Millington- ia hortensis (LC50 223.9ppm) and ethanol ex- tract was from peels of Citrus sinensis (LC50 291.69ppm) (35, 38). Extracts of A. annua, C. anthelminticum (7), Citrus sinensis, C. ternatea (35), C. gigantean (37), G. sylvestre (36) and M. hortensis (38) were less active than EF and TME of S. striata. Conclusion EF and TME of S. striata possesses better larvicidal activity against An. stephensi than other fractions. Moreover, antioxidant activ- ity of MWF, WF, and TME were higher than other fractions in both DPPH and FRAP as- says. Finally, complete phytochemical inves- tigation is suggested to reveal most effective compound in this native species. Acknowledgments This research as a thesis in Doctor of Phar- macy degree (Ms Zahra Mirzaee) has been fi- http://jad.tums.ac.ir/ J Arthropod-Borne Dis, June 2018, 12(2): 119–126 F Yousefbeyk et al.: Antioxidant and … 124 http://jad.tums.ac.ir Published Online: June 12, 2018 nancially supported by Tehran University of Medical Sciences. The authors declare that there is no con- flict of interest. References 1. Olepu S, Suryadevara PK, Rivas K, Yoko- yama K, Verlinde CL, Chakrabarti D, Van Voorhis WC, Gelb MH (2008) 2-Oxo-tetrahydro-1, 8-naphthyridines as selective inhibitors of malarial pro- tein farnesyltransferase and as anti- malarials. Bioorg Med Chem Lett. 18(2): 494–497. 2. Ghanbarzadeh S, Ghasemi S, Shayanfar A, Ebrahimi-Najafabadi H (2015) 2D- QSAR study of some 2,5-diaminoben- zophenone farnesyltransferase inhib- itors by different chemometric meth- ods. EXCLI. 14: 484–495. 3. World malaria report (2014) WHO Press GMP, World Health Organization. Ge- neva, Switzerland. World malaria re- port, WHO Press, Global Malaria Prog- ram, World Health Organization. Avail- able at: http://www.who.int/malaria/publicati ons/world_malaria_report_2014/en/. 4. Hanafi-Bojd AA, Azari-Hamidian S, Vatan- doost H, Zabihollah C (2011) Spatio- temporal distribution of malaria vec- tors (Diptera: Culicidae) across dif- ferent climatic zones of Iran. Asian Pac J Trop Dis. 4(6): 498–504. 5. Mahnaz K, Alireza F, Hassan V, Mahdi S, Reza AM, Abbas H (2012) Larvici- dal activity of essential oil and me- thanol extract of Nepeta menthoides against malaria vector Anopheles ste- phensi. Asian Pac J Trop Dis. 5(12): 962–965. 6. Zoubiri S, Baaliouamer A (2014) Poten- tiality of plants as source of insec- ticide principles. J Saudi Chem Soc. 18(6): 925–938. 7. Srivastava A, Bartarya R, Tonk S, Sri- vastava S, Kumari KM (2008) Larvi- cidal activity of an indigenous plant, Centratherum anthelminticum. J En- viron Biol. 29(5): 669–672. 8. Dı́az AMa, Abad Ma J, Fernández L, Sil- ván AM, De Santos J, Bermejo P (2004) Phenylpropanoid glycosides from Scrophularia scorodonia: in vitro anti-inflammatory activity. Life Sci. 74(20): 2515–2526. 9. Salavati P, Ramezani M, Monsef-Esfahani HR, Hajiagha R, Parsa M, Tavajohi S, Seyed Nasser Ostad (2013) Neuropro- tective effect of total and sequential extract of Scrophularia striata Boiss. in rat cerebellar granule neurons fol- lowing glutamate-induced neurotox- icity: an in-vitro study. Iran J Pharm Res. 12(2): 389–394. 10. Monsef-Esfahani HR, Hajiaghaee R, Shah- verdi AR, Khorramizadeh MR, Amini M (2010) Flavonoids, cinnamic acid and phenyl propanoid from aerial parts of Scrophularia striata. Pharm Biol. 48(3): 333–336. 11. Tasdemir D, Brun R, Franzblau SG, Sezgin Y, Çalıs I (2008) Evaluation of anti- protozoal and antimycobacterial acti- vities of the resin glycosides and the other metabolites of Scrophularia cryptophila. Phytomedicine. 15(3): 209–215. 12. Tasdemir D, Güner ND, Perozzo R, Brun R, Dönmez AA, Calıs I, Rüedi P (2005) Anti-protozoal and plasmodial FabI enzyme inhibiting metabolites of Scrophularia lepidota roots. Phyto- chemistry. 66(3): 355–362. 13. Rajkumar S, Jebanesan A (2008) Bio- activity of flavonoid compounds from Poncirus trifoliata L. (Family: Ru- taceae) against the dengue vector, Ae- des aegypti L. (Diptera: Culicidae). Parasitol Res. 104(1): 19–25. 14. Yenesew A, Twinomuhwezi H, Kabaru JM, http://jad.tums.ac.ir/ http://www.who.int/malaria/publications/world_malaria_report_2014/en/ http://www.who.int/malaria/publications/world_malaria_report_2014/en/ J Arthropod-Borne Dis, June 2018, 12(2): 119–126 F Yousefbeyk et al.: Antioxidant and … 125 http://jad.tums.ac.ir Published Online: June 12, 2018 Akala HM, Kiremire BT, Heydenreich M, Martin G. Peter, Fredrick L. Eyase, Norman C. Waters, Douglas S. Walsh (2009) Antiplasmodial and larvicidal flavonoids from Derris trifoliata. Bull Chem Soc Ethiop. 23(3): 409–414. 15. Manguro LOA, Lemmen P, Hao P (2011) Iridoid Glycosides from Underground Parts of Ajuga remota. Rec Nat Prod. 5(3): 147–157. 16. Wang Z, Kim JR, Wang M, Shu S, Ahn YJ (2012) Larvicidal activity of Cnid- ium monnieri fruit coumarins and struc- turally related compounds against in- secticide susceptible and insecticide- resistant Culex pipiens pallens and Aedes aegypti. Pest Manag Sci. 68 (7): 1041–1047. 17. Joseph C, Ndoile M, Malima R, Nkunya M (2004) Larvicidal and mosquitocidal extracts, a coumarin, isoflavonoids and pterocarpans from Neorautanenia mitis. Trans R Soc Trop Med Hyg. 98(8): 451–455. 18. Azadmehr A, Afshari A, Baradaran B, Hajiaghaee R, Rezazadeh S, Monsef- Esfahani H (2009) Suppression of nitric oxide production in activated murine peritoneal macrophages in vitro and ex vivo by Scrophularia striata ethanolic extract. J ethnopharmacol. 124(1): 166–169. 19. Rezaie-Tavirani M, Mortazavi SA, Bar- zegar M, Moghadamnia SH, Rezaee MB (2010) Study of anti cancer pro- perty of Scrophularia striata extract on the human astrocytoma cell line (1321). Iranian J Pharm Res. 9(4): 403. 20. Germinara GS, Frontera AM, De Cristofaro A, Rotundo G (2011) Insecticidal ac- tivity of different extracts from Scroph- ularia canina L. against Culex pipiens molestus Forskal (Diptera, Culicidae). J Environ Sci Health, Part B. 46(6): 473–479. 21. Pavela R (2009) Larvicidal effects of some Euro-Asiatic plants against Culex quinquefasciatus Say larvae (Diptera: Culicidae). Parasitol Res. 105(3): 887. 22. Yousfbeyk F, Esmaiili T, Pashna Z, Ho- zori Z, Ghohari A, Ostad SN, GhR Amin (2014a) Antioxidant Activity, Total Phenol and Total Anthocyanin Contents of Cornus sanguinea L subsp australis. (CA Mey.) Jáv. J Med Plant. 1(49): 69–74. 23. Moradi-Afrapoli F, Asghari B, Saeidnia S, Ajani Y, Mirjani M, Malmir M, Dolatabadi Bazaz R, Hadjiakhoondi A, Salehi P, Hamburger M, Yassa N (2012) In vitro α-glucosidase inhibitory activity of phenolic constituents from aerial parts of Polygonum hyrcani- cum. Daru. 20(1): 37. 24. Benzie IF, Strain J (1996) The ferric re- ducing ability of plasma (FRAP) as a measure of “antioxidant power”: the FRAP assay. Anal Biochem. 239(1): 70–76. 25. Yousefbeyk F, Gohari AR, Hashemi- ghahderijani Z, Ostad SN, Sourmaghi MHS, Amini M, Golfakhrabadi F, Jamalifar H, Amin G (2014b) Bio- active Terpenoids and Flavonoids from Daucus littoralis Smith subsp. Hyr- canicus Rech. f, an Endemic Species of Iran. Daru. 22(1): 12. 26. Miliauskas G, Venskutonis P, Van Beek T (2004) Screening of radical scav- enging activity of some medicinal and aromatic plant extracts. Food Chem. 85(2): 231–237. 27. Golfakhrabadi F, Ardekani MRS, Saeidnia S, Yousefbeyk F, Jamalifar H, Rame- zani N, Akbarzadeh T, Khanavi M (2016) Phytochemical analysis, antimi- crobial, antioxidant activities and to- tal phenols of Ferulago carduchorum in two vegetative stages (flower and fruit). Pak J Pharm Sci. 29(2): 623– 628. 28. Dharmagadda V, Naik S, Mittal P, Vasu- http://jad.tums.ac.ir/ J Arthropod-Borne Dis, June 2018, 12(2): 119–126 F Yousefbeyk et al.: Antioxidant and … 126 http://jad.tums.ac.ir Published Online: June 12, 2018 devan P (2005) Larvicidal activity of Tagetes patula essential oil against three mosquito species. Bioresour Tech- nol. 96(11): 1235–1240. 29. Golfakhrabadi F, Khanavi M, Ostad SN, Saeidnia S, Vatandoost H, Abai MR, Hafizi M, Yousefbeyk F, Rad YR, Baghenegadian A, Ardekani MR (2014) Biological Activities and Composition of Ferulago carduchorum Essential Oil. J Arthropod Borne Dis. 9(1): 104– 115. 30. Mahboubi M, Kazempour N, Boland Nazar AR (2013) Total phenolic, total flavo- noids, antioxidant and antimicrobial activities of Scrophularia striata Boiss Extracts. Jundishapur J Nat Pharm Prod. 8(1): 15–19. 31. Sedaghat MM, Dehkordi AS, Khanavi M, Abai MR, Mohtarami F, Vatandoost H (2011) Chemical composition and larvicidal activity of essential oil of Cupressus arizonica EL Greene against malaria vector Anopheles stephensi Liston (Diptera: Culicidae). Pharma- cogn Res. 3(2): 135. 32. Hadjiakhoondi A, Vatandoost H, Khanavi M, Abaee MR, Karami M (2005) Biochemical investigation of different extracts and larvicidal activity of Ta- getes minuta L. on Anopheles ste- phensi larvae. Iranian J Pharm Sci. 1(2): 81–84. 33. Santhoshkumar T, Rahuman AA, Raja- kumar G, Marimuthu S, Bagavan A, Jayaseelan C, Zahir AA, Elango G, Kamaraj C (2011) Synthesis of silver nanoparticles using Nelumbo nucifera leaf extract and its larvicidal activity against malaria and filariasis vectors. Parasitol Res. 108(3): 693–702. 34. Govindarajan M, Jebanesan A, Push- panathan T (2008) Larvicidal and ovicidal activity of Cassia fistula Linn. leaf extract against filarial and malarial vector mosquitoes. Parasitol Res. 102(2): 289–292. 35. Murugan K, Kumar PM, Kovendan K, Amerasan D, Subrmaniam J, Hwang JS (2012) Larvicidal, pupicidal, repel- lent and adulticidal activity of Citrus sinensis orange peel extract against Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus (Diptera: Culicidae). Parasitol Res. 111(4): 1757– 1769. 36. Khanna VG, Kannabiran K, Rajakumar G, Rahuman AA, Santhoshkumar T (2011) Biolarvicidal compound gym- nemagenol isolated from leaf extract of miracle fruit plant, Gymnema syl- vestre (Retz) Schult against malaria and filariasis vectors. Parasitol Res. 109(5): 1373–1386. 37. Kovendan K, Murugan K, Kumar KP, Panneerselvam C, Kumar PM, Amera- san D, Subramaniam J, Vincent S (2012) Mosquitocidal properties of Calotropis gigantea (Family: Asclepi- adaceae) leaf extract and bacterial in- secticide, Bacillus thuringiensis, against the mosquito vectors. Parasitol Res. 111(2): 531–544. 38. Kaushik R, Saini P (2008) Larvicidal ac- tivity of leaf extract of Millingtonia hortensis (Family: Bignoniaceae) against Anopheles stephensi, Culex quinquefasciatus and Aedes aegypti. J Vector Dis. 45(1): 66. http://jad.tums.ac.ir/