Asian Pac J Trop Biomed 2014; 4(9): 740-742 740

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Document heading doi:10.12980/APJTB.4.2014APJTB-2014-0055 2014 by the Asian Pacific Journal of Tropical Biomedicine. All rights reserved. 襃 Phytochemical screening and antioxidant activity of ethanol extract of Tithonia diversifolia (Hemsl) A. Gray dry flowers

1 1 2 1* Robson Miranda da Gama , Marcelo Guimarães , Luiz Carlos de Abreu , José Armando-Junior 1Laboratório de Pesquisa do Curso de Farmácia, Faculdade de Medicina do ABC, Avenida Príncipe de Gales, 821, Vila Príncipe de Gales, 09060- 870, Santo André, SP, Brasil

2Departamento de Saúde Materno-Infantil, Faculdade de Saúde Pública, Universidade de São Paulo, Brasil ARTICLE INFO ABSTRACT Article history: Objective: Tithonia diversifolia To evaT.lu adiversifoliate the antioxidant activity of extracts of dried flowers of ( ) ( ) Received 28 Jan 2014 Hemsl A. Gray dry flower-a shrubby belonging to the family Received in revised form 20 Mar 2014 aMethods:nd very common in Brazil, providing data to help prevent premature aging skin. Accepted 24 Apr 2014 The tests of phytochemical screening included total phenols, tannins, flavonoids, Available online 28 Jun 2014 alkaloids and saponins. The active antioxidant was determined by 2,2-diphenyl-1-picryl- hResults:ydrazyl method. T. diversifolia The phytochemical screening of dry flowers revealed the presence of Keywords: phenolic compounds (tannins, flavonoids and total phenols), while alkaloids and saponins were 50 nConclusions:ot detected. T he IC values showed a strong antioxidant activity of the plant extracts. T. A ntioxidant activity diversifolia Therefore, this study suggests the possibility of using dry flowers extracts of PTithoniahytoche mdiversifoliaical screening for the prevention of cell aging, as was shown to have significant antioxidant activity. (Hemsl) A. Gray

1. Introduction that polyphenols, especially the flavonoids possess a high antioxidant power which can protect cells against the Skin aging is commonly influenced by several factors, adverse effects of ROS[4]. such as genetic and environmental factors (UV light), Among many medicinal plant families, Asteraceae family xenobiotics, and hormonal changes. All these factors can comprises species with arboreous, shrub, herbaceous and trigger the onset of reactive oxygen species (ROS) that are liana habits and is widely distributed at tropical, subtropical chemically reactive molecules containing oxygen[1]. and tempered regions, particularly in South America, with They are formed as natural products of the normal expression in number of species, composed by some 1 535 23 000 17 [5] metabolism of oxygen and have important roles in cell genTithoniaera, diversifolia species and tribes . T. diversifolia signaling and homeostasis. However, during times of (Hemsl) A. Gray ( ) is environmental stress, ROS levels can dramatically increase a herb family (tribe ) occurring from Central that, resulting in significant damage to cell structure[2]. America to the West Indies, having been naturalized in Natural antioxidants present in plant origin protect against the tropics and also has been used as a medicinal plant these radicals and are therefore important tools in obtaining showing their anti-inflamatory[6], antimalarial[7] and many and preserving good health[3]. other biological activities. Potentially active components from fruits, herbs, roots The aims of the present study were to evaluate and leaves have been studied extensively in order to phytochemical screening andT. adiversifolialso measure the antioxidant avoid oxidative cellular events. The results suggest activity of ethanol extract of dry flowers using a 2,2-diphenyl-1-picryl-hydrazyl (DPPH) assay.

é ó *Corresponding author: Jos Armando-Junior, Laborat rio de Pesquisa do Curso á í de Farm cia, Faculdade de Medicina do ABC, Avenida Pr ncipe de Gales, 821, Vila 2. Materials and methods í é Pr ncipe de Gales, 09060-870, Santo Andr , SP, Brasil. Tel: +55(11)4993-5404 2.1. Botanic material E-mail: [email protected] ó ú çã Foundation Project: Supported by Pr -Sa de II, Funda o do ABC (Grant No. T. diversifolia 2261/2008). of were grown in the Garden of Robson Miranda da Gama et al./Asian Pac J Trop Biomed 2014; 4(9): 740-742 741 é 3. Results M P F M ABC S A ãedicinal lants, aculty of edicine of , anto ndr , S P B o aulo, razil, and flowers were collected in August 3.1. Phytochemical screening 2012 ( ) flowering season of this spec°ies . After manual collect, the material was dried at 50 C (hot air chamber) 1 ( T for week and °stored under controlled conditions dry air, T.h ediversifolia phytochemical screening test showed that the flowers dark and at 20 C). of contained several active compounds (Table 1) 50 2.2. Extract preparation . The IC of dried flower of ethanol extracts of this species and ascorbic acid are presented in Figure 1. Table 1 For extract preparation, the flowers were reduced in T. diversifolia a mill and kept in ethanol (100%) under stirring at room Phytochemical composition of ethanol extract of dry temperature for 24 h. After filtering, the extract was flowers. concentrated on the rotary evaporator attached to a vacuum Phytochemical Interference pump and then used for phytochemical screening. Total phenols + Tannins + 2.3. Phytochemical screening Flavonoids + Alkaloids - All the tests of phytochemical screening (total phenols, Saponins - tannins, flavonoids, alkaloids and saponins) followed the (+) presence; (-) absence. methodologies described at Brazilian Pharmacopea[8].

2.4. Antioxidant activity 250

) 205 80 L . 200 g / m µ The DPPH method is based on captures of DPPH by ( antioxidants, producing a decrease in absorbance at 517 nm. 150 Plant sample solutions (0.1 g/mL) were diluted to final a t i o n 100 concentrations of 250, 100, 50 and 10 mg/mL, in ethanol. 75.63 o n c e t r Ascorbic acid solutions (1.8 mg/mL) were diluted to final C µ 50 concentrations of 120, 90, 60 and 30 g/mL, in ethanol. TM A 3 L 0 04 L DPPH (S A ) bout m of a . mg/m µ igma ldrich 0 ethanol solution was added to 30 L of sample solutions Plant sample Ascorbic acid Figure 1. of different concentrations, and allowed to react at room The IC50 values (the half maximal inhibitory concentration) of T. diversifolia temperature. After 30 min, the absorbance values were ethanol extract of dry flowers and ascorbic acid. measured at 517 nm and converted into percentage ( ) 1 ( 1) antioxidant AA -using the fo-llowing equation Eq. : E 1 AA(%) 100 (A sample A blank) 100 A q. : = {[ bs bs 伊 ]/ bscontrol} µ 4. Discussion Ethanol (3.0 mL) plus plant extract solution (30 L) was used as a blank. DPPH solution (3.0 mL; 0.04 mg/mL) (3 0 ) plus ethanol . mL was used as a negative control. The In this T.st udiversifoliady, the phytochemical screening of flower positive coTMntrol was those using the standard ascorbic extract of revealed that among the substances acid (Fluka ) solutions. investigated, presence of phenolic compounds was detected The calculation equations of the analytical curves were (total phenols, tannins and flavonoids), while alkaloids and made by linear regression using the least squares method saponins were not detected. The presence of some of these of plots where the abscissa represented the concentration secondary metabolites suggests that the plant might be of of test plant extracts or ascorbic acid and the ordinate medicinal importance. represented the average percent of antioxidant activity The presence of phenolic compounds (total phenols, and calculating the linear correlation coefficient. The tannins and flavonoids) provides pharmacological activities equation 2 (Eq. 2) and equation 3 (Eq. 3) used to calculate like anti-cancer[10,11], anti-oxidant[11,12], antimicrobial[13,14], [15] [6,16] the resultsT. o f diversifoliapercent of antioxidant activity of ethanol wound-healing and anti-inflammatory , that may extract of dry flowers and ascorbic acid suggest an association to the species here investigated. reference standard, respectively. Similar results of phytochemical screening of flower extract y x R2 Eq. 2: =0.236 +1.421 5 =0.997 8 of this species were obtained by Essiett and Akpan[17], y x R2 Eq. 3: =0.621 2 +3.015 =0.995 9 differing only in the saponin presence and the result may be The half maximal inhibitory concentration (IC 50) related to the parts of the flower used to obtain the extract is a measure of the effectiveness of a compound in in the study by Essiett and Akpan that were used only the 50 [17] inhibiting aT.n tidiversifoliaoxidant activity. The IC values of ethanol petals of flowers , while in present study were obtained by extract of dry flowers and ascorbic acid the whole flower. 2 3 reference standard were calculated by Eq. and Eq. , The theory of aging skin by the action of free radicina lvivos is respectively[9]. based on the failure mechanism of natural antioxidant Robson Miranda da Gama et al./Asian Pac J Trop Biomed 2014; 4(9): 740-742 742 in vitro - catechins in the prevention of the metabolic syndrome a review. and since studies suggest a correlation between the Phytochemistry 70 2009; : 11-24. aging process and reducing enzymatic and non-enzymatic á á agents, with a consequent increasing level of ROS[18]. [5] J tem-L sser A, Ricardi MS, Adamo G. Herbal traditional medicine of Venezuelan Andes: an ethnopharmacological study. One of the most widely used natural antioxidants studied Phytother Res 12 - is ascorbic acid that eliminates most ROS due to the 1998; : S53 S59. [6] C apelari-Oliveira P, Paula CA, Rezende SA, Campos FT, Grabe- oxidation of ascorbate to monodehydroascorbate and then ã Guimar es A, Lombardi JA, et al. Anti-inflammatory activity to dehydroascorbate and has other functions to maintain the Lychnophora passeria “ ” J of , Asteracea (Brazilian Arnica ). normal physiologic state in humans. In the skin, ascorbic Ethnopharmacol 135 acid is a cofactor required for the enzymatic activity of 2011; (2): 393-398. [7] L acerda AM, Modolo AK, Matias RC, Pistori H, Yano M, Roel prolyl hydroxylase, which hydroxylates prolyl resulting in Rev Bras [19] AR, et al. Screening of plants with potential phototoxic. procollagen and elastin . Farm 92 2011; (4): 352-355. Bogdan Allemann and Baumann revised the use of ê â á [8] A g ncia Nacional de Vigil ncia Sanit ria. Brazilian Pharmacopea. other antioxidants in skin care formulations and found í ê â á that a 3-month daily regimen of topical using of ascorbic 5th ed. Bras lia: Ag ncia Nacional de Vigil ncia Sanit ria; 2010. acid provided objective and subjective improvement in [Online] Available from: http://www.anvisa.gov.br/hotsite/cd_ photodamaged facial skin[20]. farmacopeia/pdf/Volume%201.pdf [Accessed on 12th March, 2014] I Portuguese. t is common consensus’ that the cellular aging process ã can be prevented by plants phenolic substances, which has [9] M ensor LL, Menezes FS, Leit o GG, Reis AS, dos Santos TC, Coube CS, et al. Screening of Brazilian plant extracts for antioxidant motivated the investigation of these plant metabolites and Phytother Res [21] activity by use of DPPH free radical method. 2001; their possible action in the prevention of cellular aging . T. 15 50 (2): 127-130. diversifolia The result shows the values of IC of ethanol extract of [10] L in Y, Shi R, Wang X, Shen HM. 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Afr J Biotechnol 5 Finally, considering the results obtained, as future medicinal plants. 2006; (11): 1142-1145. perspectives, we intend to evaluate some biological [13] S engul M, Yildiz H, Gungor N, Cetin B, Eser Z, Ercisli S. Total phenolic content, antioxidant and antimicrobial activities of some activities, such as wound-healing, anti-inflammatory, Pak J Pharm Sci 22 antimicrobial and anti-cancer activity, as well as quantify medicinal plants. 2009; (1): 102-106. the main phytochemicals present in extracts. [14] H endra R, Ahmad S, Sukari A, Shukor MY, Oskoueian E. Flavonoid analyses and antimicrobial activity of various parts of Phaleria macrocarpa Int J Mol Sci 12 (Scheff.) Boerl fruit. 2011; : Conflict of interest statement 3422-3431. Catharanthus roseus [15] N ayak BS, Pinto Pereira LM. flower extract BMC has wound-healing activity in Sprague Dawley rats. We declare that we have no conflict of interest. Complement Altern Med 6 2006; : 41. 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