Pluchea Indica Less) LEAVES EXTRACT AS ANTIOXIDANT and ANTI WARMED OVER FLAVOR (WOF) of DUCK MEAT
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POTENCY OF BELUNTAS (Pluchea indica Less) LEAVES EXTRACT AS ANTIOXIDANT AND ANTI WARMED OVER FLAVOR (WOF) OF DUCK MEAT Paini Sri Widyawati1, Tarsisius Dwi Budianta, Fenny Anggraeni Kusuma, Evelyn Livia Wijaya, Dian Ivana Yaunatan, Ribka Stefanie Wongso Faculty of Agricultural Technology, Widya Mandala Catholic University of Surabaya Jl. Dinoyo 42‐44 Surabaya, 60265, Indonesia Correspondent author: : [email protected] Abstract Beluntas (Pluchea indica Less) is a herb plant used as a traditional medicine or eaten in fresh form. There are phytochemical compounds such as essential oils, flavonoids, phenolics, tannins, saponins, phenols hydroquinone, and cardiac glycosides compounds of beluntas leaves that cause it having potential as anti‐ oxidant. Difference of solvent polarity can cause concentration and composition of phytochemical com‐ pounds in extract differed, therefore this study was conducted to determine the potential for beluntas leaves extracts (water, methanol, ethanol, ethyl acetate, and hexane) and fractions (water, ethyl acetate, and n‐butanol) as an antioxidant and antiwarmed over flavor (WOF) in duck meat during storage. The results showed that the methanol extract of the beluntas leaves (EMB) of the most potential as a source of antioxidants because the concentration and composition of phytochemical compounds, total phenols and total flavonoids than the water, ethanol, ethyl acetate, and hexane extracts. Furthermore EMB was frac‐ tionated by difference of solvent polarity (ethyl acetate, water, n‐butanol). Test showed that EMB antioxi‐ dant capacity and its fractions had the difference in the ability of antioxidant compounds in the EMB and each fraction in different test systems. EMB had the potency to scavenge superoxide radicals, reduce iron ions, and inhibit bleaching of linoleic acid‐β‐carotene system. Ethyl acetate fraction (FEA) had the po‐ tency to scavenge superoxide radicals, reduce iron ions, chelate of iron ions and haemoglobin (Hb), thus FEA was effective as antiwarmed over flavor (WOF) in duck meat, which protected linoleic acid, decreased of TBARS and hexanal. Keywords: beluntas (Pluchea indica Less), antioxidant, antiwarmed over flavor, duck meat Abbreviations: pene, phenylpropanoid, benzoid, alkanes, sterol, 2 EMB :Beluntas methanol extract ‐(prop‐1‐unyl)‐ 5‐ 5, 6‐dihydroxy hexa‐1,3‐diunyl)‐ FEA :Ethyl acetate fraction thiophene, (‐)‐catechin, phenol hydroquinone, DPPH : 2,2‐diphenyl‐1‐picrylhydrazyl radical saponin, tannin, and alkaloid, flavonol (quercetin, AT : Alpha tocopherol kaempherol, myricetin, luteoline, apigenine) WOF : Warmed over flavor Hb : Haemoglobin (Andarwulan et al., 2010). Generally, phytochemi‐ AC :Control absorbance cal compounds, especially phenolic in plants have AS : Sample absorbance free structure or glycoside and ester bonds UV‐Vis : Ultra violet‐visible (Dehkharghanian et al., 2010). There are hydroxyl GAE : Gallic acid equivalent groups at phenolic structure caused these com‐ CE : Cathecin equivalent pounds having polar properties (Lai et al., 2009). IC : Inhibition concentration Water, ethanol, and methanol are polar solvent TBARS : Thiobarbituric acid reactive substances that can extract glycoside structure of phenolic BHT : Butyl hydroxyl toluene compounds. Ethyl acetate has potentially to ex‐ DM : Fresh duck meat DP : Cooking duck meat tract alkaloid, aglycon, and glycoside compounds DPSP : Warmed stored cooking duck meat (Houghton and Raman, 1998), phenolic com‐ MDA :Malondialdehyde pounds with low until high molecular weight (Mariod et al., 2010), and then hexanes can extract 1. Introductions lignin, wax, lipid, aglycon, sterol, and terpenoid Beluntas (Pluchea indica Less), herb plant is a (Houghton and Raman, 1998). Previous research species in the family Asteraceae, used as a tradi‐ had found that aqueous extract of beluntas leaves tional medicine or eathen in fresh form can scavenge 2,2‐diphenyl‐1‐picrylhydrazyl (Ardiansyah et al. 2003). Beluntas leaves contain (DPPH) free radical, superoxide and hydroxyl phytochemical compounds such as lignin, ter‐ radicals, iron reducing power, and iron ion che‐ 81 lating. Ethanolic extract of beluntas leaves can 2.2. Preparation of beluntas leaves extracts scavenge 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) and its fractions free radical and 2,2’‐azino‐bis‐ (3‐ethyl benzo thi‐ Extraction of beluntas leaves was based on azoline‐ 6‐sulphonic acid (ABTS) cation radical, Dorman dan Hiltunen (2004) modification iron ion reducing power, and inhibit bleaching method. 1‐6 segment beluntas leaves sorted, linoleic acid‐β‐carotene system (Andarwulan et washed, and dried at ambient temperature for a al., 2010). Methanol is effective to extract phenolic week were milled with 40 mesh. And then belun‐ compounds with low molecular weight having tas leaves flour was analysed moisture content medium polarity and results the highest yield and and extracted with different solvent polarity i.e. antioxidant activity (Chan et al., 2007). Neverthe‐ water, methanol, ethanol, ethyl acetate, and hex‐ less until now ethyl acetate and hexanes haven’t anes with 1:15 b/v ratio for 3 hours at boiling known antioxidant activity. Fractionation of be‐ point. Extract was evaporated by rotary evapora‐ luntas leaves extract with several solvent different tor at 40oC. And then extract was analysed yield, polarity, such as ethyl acetate, water, and n‐ phytochemical compounds, and antioxidant activ‐ butanol is done to result yield with different anti‐ ity. Extract having the highest antioxidant activity oxidant compound composition (Lai et al., 2009), was fractionated by different solvent polarity so that can resulted extract or fraction the most (ethyl acetate, n‐butanol, and water). And then potentially as antioxidant source. Each solvent fraction was evaporated at 40oC and analysed can solve phytochemical compound with different yield, phytochemical compounds, and antioxidant composition. n‐butanol can extract polar com‐ activity. pounds, i.e. glycoside, aglycon, dan gula (Liu dkk., 2.3. Moisture Content 2011). WOF (Warmed over flavor) is flavor degra‐ Dried beluntas leaves were measured mois‐ dation at food product because of food product ture content based on AOAC (1990) method. warmed again after they are stored in refrigerator 2.4. Yield at 4‐5oC for 48 hours (Jayathilakan et al., 2007). Yield was determined by Ljubuncic et al. WOF can be happened by lipid oxidation at meat (2005) method that measured by compared with and heme protein denaturation that can acceler‐ beluntas leaves extract or its fractions weight and ate lipid oxidation (Masqood and Benjakul, 2011). dried beluntas leaves. Duck meat can be oxidized to result WOF be‐ 2.5. Assay of Phytochemical Compounds cause it contains high lipid at adipose tissue. Until Phytochemical was determined based on Har‐ now antioxidant capacity of beluntas leaves ex‐ borne (1996) method. tract and its fractions haven’t known to inhibit 2.6. Assay of Total Phenol WOF so that it is studied. Beluntas leaves extract Total phenol was measured by Manian et al. (EMB) and its fractions can reduce synthetic anti‐ (2008) method using Folin ciocalteus phenol re‐ oxidant using at food product because of carcino‐ agent. Data was stated as mg gallic acid equiva‐ genic (Valentao et al., 2010). The research was lent (GAE) per 100 gram sample dry base. Solution done to study the potency of beluntas leaves ex‐ absorbance was measured by spectrophotometer tracts (water, methanol, ethanol, ethyl acetate, UV‐Vis at λ 760 nm. and hexanes) and its fractions (water, ethyl ace‐ 2.7. Assay of Total Flavonoid tate, and n‐butanol) as antioxidant and anti Total flavonoid was determined by Manian et warmed over flavor at duct meat for storage. al. (2008) method using NaNO2‐AlCl3‐NaOH re‐ agent, data was stated by milligram catechin 2. Methods equivalent (CE) per 100 gram sampel dry base. 2.1. Materials Solution absorbance was measured by spectro‐ Beluntas leaves are harvested at Dramaga photometer UV‐Vis at λ 510 nm. area, Bogor and Sukolilo area, Surabaya. Green tea 2.8. Assay of DPPH Free Radical Scavenging is purchased at Tea Factory in Singapura (Lim Activity Lam Thye PTE, LTD). Dried Rosemary is pur‐ DPPH free radical scaveging activity was chased at Cold storage supermarket in Holland measured based on Manian et al. (2008) method Evanue, Singapura. Chicken blood Darah ayam is modification. Decreasing of purple color solution cultivated from Laladon traditional market, Bo‐ was determined by spectrophotometer UV‐Vis at gor. Chemical is used Analytical grade from λ 517 nm after reaction occurred 30 min. Prosen‐ Sigma, Merck, JT Beaker, and Riedel‐de Haen, tase of inhibition (%) was calculated based on = except ethanol (PT Brataco) and aquadest from [(AC‐AS)/AC] x 100%, where AC = control absorb‐ Microbiology laboratory, SEAFAST‐PAU‐IPB. ance, AS = sample absorbance. 82 2.9. Assay of Superoxide Radical Scavenging AS)/AC] x 100%, where AC = control absorbance, Activity AS = sample absorbance. Superoxide radical scavenging activity was 2.15. Assay of Linoleic acid‐β‐Carotene Bleach‐ determined by Manian et al. (2008) method. Su‐ ing Capacity peroxide radical was resulted by NBT‐NADH‐PMS Linoleic acid‐β‐carotene bleaching capacity reaction. Solution absorbance was measured by was measured by Manian et al. (2008) method. spectrophotometer UV‐Vis at λ 560 nm. Prosen‐ Decreasing of orange color solution because of β‐ tase of inhibition (%) was calculated by = [(AC‐ carotene oxidation was detected by spectropho‐ AS)/AC] x 100%, where AC = control