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Fabaceae) from Turkey Antioxidant capacity and fatty acid composition of different parts of Adenocarpus complicatus (Fabaceae) from Turkey Adnan Berber1, Gokhan Zengin2, Abdurrahman Aktumsek2, Murad Aydin Sanda2 & Tuna Uysal2 1. Necmettin Erbakan University, Seydisehir Ahmet Cengiz Engineering Faculty, Department of Mechanical Engineering, Konya, Turkey; [email protected] 2. Selcuk University, Science Faculty, Department of Biology, Konya, Turkey; [email protected], [email protected], [email protected], [email protected] Received 25-II-2013. Corrected 30-VII-2013. Accepted 28-VIII-2013. Abstract: Adenocarpus complicatus is distributed throughout the Anatolian peninsula and is widely used for human and animal nutrition. The purpose of this work was to study the antioxidant properties and fatty acid composition of different parts of this plant (fruits and mixed materials). The species was collected from Golyuzu village of the Seydisehir district near Konya province, Turkey. Fruit and mixed parts obtained from this species were ground and a 15g sample was used to prepare methanolic extracts. Powdered plant samples were extracted with 100mL methanol in a mechanical shaker. The obtained extracts were filtered and concentrated to dryness under reduced pressure and were subsequently stored at -20°C. Antioxidant components, namely total phenolic and flavonoid content, were detected for each extract using spectrophotometric methods. Antioxidant capacity was evaluated by various assays including phosphomolybdenum, DPPH free radical scavenging capacity, metal chelating activity, and ferric and cupric ion reducing power. The fatty acid profiles of plant parts were also deter- mined by using gas chromatography. The total phenolic content of fruit (36.21mgGAE/g) was higher than that of mixed materials (13.79mgGAE/g). The methanolic extract of mixed material had higher amounts of flavonoid than fruit extract. The free radical scavenging activity of extracts was expressed as IC50 value (µg/mL) (amount required to inhibit DPPH radical formation by 50%). The lower IC50 value reflects better free radical scavenging action. The radical scavenging activity of the samples was compared with BHT, it showed the mixed material to be almost two times more potent than the fruit extract. However, BHT is an excellent free radical scavenger with an IC50 of 34.061µg/mL. The ferric and cupric reducing power potentials of the extracts were expressed as EC50 value (the effective concentration at which the absorbance was 0.5). Fruit extract exhibited strong ferric reducing power with an EC50 of 871.25µg/mL. The metal chelating activity of the extracts increased with concentration. Chelating effect was 83.60% for fruit extract at 1mg/mL concentration. Oil content of fruit and mixed parts were detected as 6.71 and 6.14%, respectively. A total of 32 fatty acids were found in the oil. Essential fatty acids (lin- oleic and α-linolenic acid) were identified as the most abundant fatty acids in the oil. These results demonstrated that this plant species can be considered as an alternative to synthetic antioxidants. Likewise, the oil obtained from the plant can be used as a source of essential fatty acids for food and pharmacological applications. Rev. Biol. Trop. 62 (1): 337-346. Epub 2014 March 01. Key words: Adenocarpus complicatus, antioxidant capacity, fatty acid composition, total flavonoid content, total phenolic content. Free radicals are an atom or a group by normal metabolic processes or exogenous of atoms with an unpaired electron and are factors such as ultraviolet light and cigarette generally known as reactive oxygen species, smoke. Overproduction of these radicals is which include hydrogen peroxide, hydroxyl considered to be the main contributor to oxida- radical, nitric oxide, peroxynitrite, singlet tive stress (Ellnain-Wojtaszek, Kruczynski & oxygen, peroxyl radical and superoxide anion Kasprzak, 2003). Oxidative stress is involved (Halliwell, 2001). These radicals are generated in many acute and chronic diseases including Rev. Biol. Trop. (Int. J. Trop. Biol. ISSN-0034-7744) Vol. 62 (1): 337-346, March 2014 337 cancer, cardiovascular pathologies and neuro- potential and fatty acid composition for Ade- degenerative diseases (Bouayed et al., 2007). nocarpus complicatus. Thus, the purpose of The human body has multiple mechanisms, this work was to investigate the total pheno- especially enzymatic systems, to protect cellu- lic and total flavonoid contents, the antioxi- lar components against damage-inducing dant properties and the fatty acid profile of reactive oxygen species (Anderson, 1999). Adenocarpus complicatus. However, the defense system may not be enough for persistent oxidative stress. Hence, MATERIALS AND METHODS intake of exogenous antioxidants is required in order to balance the reactive oxygen species Plant material: Root, stem, leaves, flowers in the human body. Antioxidants are known to and pods were collected and mixed from speci- play an important role in protection against oxi- mens obtained from a single location. dative damage. Epidemiological studies have Fruits: Particularly, pods are selected and shown that food rich in antioxidants plays an separately analyzed because of their dense essential role in the prevention of certain disea- and visible oil content. For this reason, mature ses such as cancer and cardiovascular diseases fruits were collected from same location. (Ismail, Marjan & Foong 2004). However, Both mixed and fruit materials were collec- synthetic antioxidants such as BHA (butylated ted from Golyuzu village of the Seydisehir hydroxyanisole), BHT (butylated hydroxyto- district near Konya province (37°20’49.73” N luene), TBHQ (tertiary butylhydroquinone) - 31°55’22.68” E) between July and August and PG (propyl gallate) have been restricted 2009. The plants were identified by Dr. Tuna for use in food industry because of some side Uysal from Section of Molecular Systematics, effects (Namiki, 1990). Therefore, there is Department of Biology, Science Faculty, Selcuk great interest in finding new and safe antio- University. The voucher specimens have been xidants from natural sources. Several plant deposited in KNYA herbarium at the department extracts and phytochemicals such as flavo- of Biology at Selcuk University (Voucher Num- noids, tannins and other phenolic compounds ber: KNYA 14 062, Collected number: Uysal have strong antioxidant potential and are used 1639, Sanda and Berber). as additives in the food industry (Salah, Miller, Paganga, Tijburg, Bolwell & Rice-Evans 1995; Preparation of methanolic extracts: The Van Acker et al., 1996). different plant materials were dried under shade Fabaceae is a family of flowering plants at room temperature and powdered to a fine comprising about 269 genera and 5 100 species grain using a laboratory mill (Simsek Labor- and is one of the largest plant families in the technic, HD-702). For methanolic extraction, world. In Turkey, it is represented by 68 genera powdered plant samples (15g) were extracted and more than 900 species (Davis, 1970). Ade- with 100mL methanol in a mechanical shaker nocarpus is a genus of this family, and belongs (Topo, MS300-HS) for 8h at room temperature. to the sub family of Faboideae. The genus is The extracts were filtered and evaporated under solely represented with the perennial Adeno- vacuum to dryness, using a rotary evaporator carpus complicatus in Turkey. The species (Heidolph Laborota 4000). The extracts were is mainly distributed in Western, Central and stored in the dark at -20°C. Southern Anatolia (Mill & Tan, 1988). Some Fabaceae species have been inves- Chemicals: Potassium ferricyanide, ferric tigated for their antioxidant capacity and fatty chloride, Folin-Ciocalteu’s phenol reagent, tri- acid composition (Akpinar, Akpinar & Turko- chloroacetic acid, methanol, butylated hydro- glu, 2001; Bagci, 2006; Tung, Wu, Hsieh, xytoluene (BHT) and butylated hydroxyanisole Chen & Chang, 2009; Custodio et al., 2009). (BHA) were purchased from Merck (Darmstadt, However, there is no data about antioxidant Germany). 2, 2-diphenyl-1-picrylhydrazyl 338 Rev. Biol. Trop. (Int. J. Trop. Biol. ISSN-0034-7744) Vol. 62 (1): 337-346, March 2014 (DPPH), ferrozine (3-(2-pyridyl)-5-6-diphenly- mixture was incubated at 95°C for 90 minutes. 1,2,4-triazine-4’,4”-disulfonic acid sodium The absorbance of the solution was then mea- salt) and FeCl2 were purchased from Sigma sured at 695nm against a blank. Results were Chemical Co. (Sigma-Aldrich GmbH, Stern- expressed as milligrams ascorbic acid (AE) and heim, Germany). All chemicals and solvents trolox equivalents (TE) per gram of extracts. were analytical grade. Scavenging activity on DPPH (2, Assay for total phenolics: The total phe- 2-diphenyl-1-picrylhydrazyl) radical: The nolic content of the extracts was determined free radical scavenging activity of plant extracts using the Folin-Ciocalteu reagent (Slinkard was determined with the method described & Singleton, 1977). A sample of 0.2mL of by Sarikurkcu, Tepe, Daferera, Polissiou & extract solution (1mg/mL) were mixed with Harmandar (2008) with slight modifications. A 1mL Folin-Ciocalteu reagent and 2mL Na2CO3 total of 0.5mL of various concentrations of the (7.5%). The final volume was brought up to methanol extracts was added with 3mL (6.10- 7mL with deionized water. After 2h of incu- 5M) of a methanol solution of DPPH. The solu- bation at room temperature, the absorbance tion was incubated for 30min in the dark at room was measured at 765nm using a spectrophoto-
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