Abstract. Ramiréz-Aristizabal, L. S. * ; Ortiz, A. ; Restrepo-Aristizabal, M.F.; Salinas-Villada, J.F. Universidad Tecnológica de Pereira,Colombia

Transcription

1 Evlution of the ntioxidnt cpcity nd content of polyphenols obtined from te (Cmelli sinensis) of four brnds sold in Colombi by extrction t room temperture Rmiréz-Aristizbl, L. S. * ; Ortiz, A. ; Restrepo-Aristizbl, M.F.; Slins-Villd, J.F. Universidd Tecnológic de Pereir,Colombi * lurmire@utp.edu.co Abstrct Herbl medicines hve long been used to tret chronic diseses such s cncer, neurodegenertion nd dibetes, usully in the form of herbl tes, lso clled tisnes [1]. Te, second only to wter, is the most highly consumed beverge worldwide nd its medicinl nd helth properties, long known to erly Chinese civiliztions, hve been widely explored [2]. The consumption of green te in Colombi is recent trend nd the mrket is continuously growing, then the most common commercilly vilble types of green te were tested in this study; Orientl, Lipton, Hindú nd Jibel. The min objective of this study ws to determine nd compre the mount of polyphenols present in te smples considering extrction in wter t room temperture nd stirring every 30 seconds for 5 minutes. The ntioxidnt cpcity ws determined by the 1,1- diphenyl-2-picrylhydrzyl (DPPH) free-rdicl scvenging ssy nd the oxygen rdicl bsorbnce cpcity (ORAC) ssy. Concentrtions rnging from ± 0.98 to ± 0.86 mg Trolox equivlent / g dry smple for DPPH ssy nd ± 1.31 to ± 2.05 mg Trolox equivlent / g dry simple were determined for ORAC ssy. It ws lso found tht the mount of totl phenols in te smples rnging from 2.53 ± 0.25 to ± 0.53 mg gllic cid equivlent (GAE) / g dry smple nd totl flvonoid content concentrtion ws obtined from 2.67 ± 0.20 to 7.08 ± 0.38 mg ctechin equivlent / g dry simple. The ntioxidnt ctivity nd totl flvonoid content were highly correlted for both DPPH (r 2 = ) s ORAC (r 2 = ). Te extrcts from the Orientl brnd hd the highest polyphenol content showing greter biologicl ctivity, contrry to Jibel te brnd which recorded the lowest concentrtions in ll nlyzes. Key words: Antioxidnt ctivity, cmelli sinensis, green te, room temperture.

2 Introduction Green te (Cmelli sinensis) is well known for vrious helth benefits ssocited with risk reduction of wide rnge of chronic diseses, such s cncer, dibetes, nd crdiovsculr diseses [3]. In the pst few yers, specil ttention hs been pid to the ntioxidnt ctivities of the polyphenolic compounds present in green te due to their phrmceuticl properties. The performed studies report tht green te extrct (GTE) shows mny helth beneficil properties, prticulrly ginst the dmge cused by pollution, stress, cigrette smoke nd other toxins, prevention of crdiovsculr diseses, cncer, dibetes [4]. Te is rich in polyphenolic compounds, nmely the ctechins, (+)-ctechin (C), ( )-epictechin (EC), ( )- epictechin gllte (ECG), ( )-epiglloctechin (EGC), nd ( )-epiglloctechin gllte (EGCG) re thought responsible for such beneficil effects [5]. The methods of prepring the beverge lso vry worldwide: in Chin, te leves re steeped in hot wter (70 80 C for green te) for s, nd the sme te leves re usully repetedly steeped (seven times). The Jpnese usully prepre green te by steeping leves in hot wter for bout 2 min nd using them for 2 3 infusions. In the United Kingdom, Irelnd nd in Cnd, blck te is mostly prepred using boiling wter nd consumed with milk nd often sugr. Americns re lrge consumers of iced te which is mde from hot te cooled with ice. In recent times in Tiwn, especilly in summer, cold wter (4 or 25 C) steeping is new populr wy for mking te [6]. Severl studies hve been done on the vrious brewing conditions to mximise the extrction of these components from green te. The results hve defined optiml wter brewing conditions including the temperture being mintined t 80 C for 30 min nd rtio of te to wter of 1:20 g/ml [7]. However, these optimistion studies focused on loose-lef green te nd were crried out under lbortory conditions. These conditions re very different from household brewing hbits, where te is simply brewed in boiled wter nd left t room temperture for short time (3 min) before being consumed [8]. This study is imed to investigte whether the ntioxidnt cpcity of different types of te could be ffected by steeping te bgs in cold wter during short time of steeping nd tking into ccount household hbits. Methods Te Smples A set of four commercil brnds of green te (Orientl, Lipton, Hindú) nd combintions of green te with certin fruits nd romtic flowers (Jibel with Hibiscus sbdriff nd lemon). They were used in the experiment for comprison of cold te infusions. These te smples were purchsed t the locl supermrket nd nlyzed in triplicte ccording with the temperture of infusion. Smple preprtion In order to simulte beverge brewing, ech brnd of te were prepred using n queous extrction procedure, to study the effect of different extrction condition on the totl phenols, flvonoids nd ntioxidnt ctivity. Cold te infusions were prepred by dding 150 ml of potble wter t room temperture to tebg tht ws weighed (tble 1) nd leving the infusions to stnd t room temperture (27 28 C) for 5 minutes, gitting mnully every minute nd controlling it s temperture every 30 seconds. See figure 1. Three extrctions of ech te brnds (Orientl, Lipton, Hindú nd Jibel) were performed for three consecutive dys for cold extrct. Just three extrctions for ech of the four brnds were retined by type of infusion. Smples were diluted t ppm in distilled wter nd stored t 4 C for subsequent nlysis. Totl phenol content (TPC) Totl phenol content of the different te smples ws determined ccording to the Folin-Cioclteu method described by V. L. Sigleton, R. Orthofer nd R.M. Lmuel- Rventós [9]. The smples were diluted 1:10 in distilled wter, 160 μl of ech smple, nd stndr were dded to polystyrene cells of 3000 μl. Then 1420 μl Folin regent (1:10) nd 1420 μl of N2CO3 7.5% were dded nd homogenized, After mixing, the smples were left for 120 minutes t room temperture in the drk, the bsorbnce ws red t 758 nm ccording with the spectrl scnning (SHIMADZU UV-1700 spectrophotometer). All blnks contined distilled wter nd te, the phenolic compounds were quntified using clibrtion curves of Gllic cid nd expressed s mg gllic cid equivlent / g smple (mg GAE/g smple). Kinetics of the rection ws conducted for period of two hours with te smples nd stndrds to determine the stbility of the complex formed ccording with time. Totl flvonoid content (TFC) The TFC ws determined ccording to D.O Kim, K.W. Lee, H.J. Lee nd C.Y. Lee [10]. The solution is mixture of

3 PhOL sodium nitrite solution followed by luminum thrichloride with sodium hydroxide nd distilled wter, which is mixed well nd the bsorbnce is mesured ginst freshly prepred regent blnk. The extrct ws rected with luminum chloride for determintion of flvonoid content. A pink color ppers fter few minutes which indicte the presence of flvonoids ccording with J. Zhishen, T. Mengcheng, nd W. Jinming [11]. Briefly, 600μL of distilled wter ws dded to 150 μl of ech smple of hot nd cold extrction te nd stock solution of ctechin intro polystyrene cells of 3000 μl, 10 μl of 5% sodium nitrte (NNO3) ws dded. The mixture ws incubted in the drk for 5 minutes. A 45 μl volume of luminum chloride (AlCl3, 10%) ws dded to the mixture nd incubted for further 6 minutes in the drk. A 300 μl of NOH, 1M, ws dded to the resulting mixture followed by ddition of 360 μl of distilled wter. The bsorbnce ws red t 505 nm ccording with the spectrl scnning (SHIMADZU UV-1700 spectrophotometer). The smples were diluted 2:10 in distilled wter, ech blnk contins te extrct nd distilled wter. The flvonoids contents were quntified using clibrtion curves of Ctechin nd expressed s mg Ctechin equivlent/g smple (mg CE/g smple). Kinetics of the rection ws conducted for period of one hour with te smples nd stndrds to determine the stbility of the complex formed ccording with time. Antioxidnt ctivity DPPH ssy DPPH ssy ws crried out in photometric cells of 3mL tking into ccount chnges in the method of O. P. Shrm [12]. A volume of 300 µl te cold extrct (ll brnds were 10 diluted times) nd/ or trolox stndrd with their respective controls; positive control ws hydroquinone 1000 ppm; negtive control ws methnol 96%, were dded to photometric cells followed by 2700 ml of methnol solution of DPPH t 30 ppm except the blnk smple. The smples were incubted in the drk t room temperture for 30 minutes (ccording to the kinetics of the rection ws determined for the methnolic solution of DPPH nd ech of the te smples nlyzed: Orientl, Hindú, Jibel nd Lipton) nd red t wvelength of 517 nm by spectrophotometer (SHIMADZU UV-1700 spectrophotometer). The scvening cpcity ws clculted s: % A.A= [A c (-) A te /Ac (-)]*100, where Ac (-) is the bsorbnce of the control nd Ate is the bsorbnce of the tested smple. Trolox ws used s stndrd. Free rdicl scvenging cpcities of te were expressed s mg Trolox Equivlent/ g smple. (mg TE/ g smple). Kinetics of the rection ws conducted for period of one hour with te smples nd stndrds to determine the stbility of the complex formed ccording with time. ORAC ssy The ntioxidnt ctivity of te smples were lso nlyzed by ORAC (Oxigen Rdicl Absorbnce Cpcity) ccording to the method of K. M. Gillespie [13]. Dilutions of te extrcts rnging from 80 to 50 diluted times in PBS t 75 mm hving te cold extrcts (Orientl nd Lipton: 80 diluted times; Hindú nd Jibel: 50 diluted times). The diluted smples were dded to 96 wells with solid white plte followed by 187 ul of 80 nm fluorescein diluted in PBS (phosphte buffered sline) t 75 mm. After 15 minutes incubtion in the drk t 37 C solution of 140 mm AAPH diluted in 75 mm PBS is prepred nd dded to ech well of the plte; AAPH is responsible for strting the decy of fluorescein so it is importnt to hve fst nd strict control of the ddition, recording dt every 120 seconds until decy of fluorescein, using n emission wvelength of 493 nm nd filter excittion 515 nm using fluorescence spectrophotometer (Vrin, Cry Eclipse, version 1.1 (135)). The net AUC (re under the fluorescence decy curve) for ech smple/stndrd ws obtined by subtrcting the re of the blnk smple (PBS). Antioxidnt ctivity ws expressed s mg Trolox equivlent/ g smple (mg TE/g smple) using the liner regression vlue obtined from the trolox clibrtion curve. Results Totl phenol content (TPC) The totl phenolic content of the 4 brnds green te re shown in figure 2 nd tble 2, were quntified using clibrtion curves of gllic cid (1-30 mg/l) performed every dy of the ssy. The totl phenolic compounds were found between mg gllic cid Equivlent /g smple. The highest levels ws mesured in Orientl brnd (14.63 ± 0.53 mg equivlent gllic cid / g smple),

4 PhOL similr mounts were lso obtined in Lipton (10.38 ± 0.55 mg gllic cid equivlent / g smple) nd Hindú (9.89 ± 0.66 mg gllic cid equivlent / g smple), while Jibel contined the lowest mount (2.53 mg gllic cid equivlent / g smple). Totl flvonoid content. (TFC) The totl flvonoid content of the te extrct were low compred with TPC, nd rnged from 2.67 ± ± 0.38mg ctechin equivlent / g smple, (see figure 3 nd tble 2) nd quntified using clibrtion curve of ctechin (1-30 mg/l) performed every dy of the ssy. Orientl nd Hindú hd the highest levels compred to other brnds (Orientl: 7.08 ± 0.38 mg ctechin equivlent / g smple, Hindú: 6.71 ± 0.36 mg ctechin equivlent / g smple), for Lipton brnd ws obtined content of 4.87 ± 0.26 mg ctechin equivlent/ g smple, while Jibel contined the lowest mount (2.67 ± 0.20 mg ctechin equivlent / g smple). Antioxidnt Activity The ntioxidnt ctivity of the te infusions ws evluted using two independent ssys, DPPH nd ORAC. A clibrtion curve of trolox ( um) llowed to compre ntioxidnt ctivity in different brnds of te expressed s μmol Trolox equivlent / g dry smple. The results obtined from DPPH ssy reported in the figure 4 nd tble 2, shown vlues rnged from ± ± 0.86 mg Trolox equivlent / g dry smple, similr to the vlues obtined by ORAC ssy reported in the figure 5, which hve rnge from ± ± 2.05 mg Trolox equivlent / g dry smple. The Orientl nd Hindú brnd te hd the highest DPPH nd ORAC vlues while the Jibel brnd showed the lowest. Discussion The degree of oxidtion of the leves defines the type of te: white, yellow, green, oolong, pu-erh nd blck te. Green te is the lest processed, resulting from quick drying of the fresh leves, with miniml oxidtion, which mke it richer in bioctive polyphenols comprtively to more processed tes, where these compounds re degrded during the process. The consumption of te, especilly green te, hs severl well-estblished helth benefits, nmely the reduction of the incidence of oxidtive stress relted diseses nd crdiovsculr disorders, for exmple [14]. The helth benefits of green te re minly ttributed to their high phenolic content, which mke these beverges one of the mjor sources of helth promoting polyphenols in our diet [1]. Extrction is the initil nd the most importnt step in the recovery nd purifiction of bioctive compounds from plnt mterils. In generl, the conventionl techniques for green te extrction re heting, boiling, Soxhlet extrction nd cold extrction, which re ll limited by long extrction periods nd low extrction efficiency [15]. The results obtined from the nlysis of the ntioxidnt ctivity of te infusions by E. Dmini nd T. Bcchetti [16] showed tht green te exhibited greter ctivity when steeped for 2 h in wter t room temperture. The different ffinities of the extrction solvents for totl te lef constituents in terms of their different extrction conditions, such s polrity of extrcting solvents, nd temperture ply n importnt role while investigting the phytochemicl profile nd ntioxidnt functions of te [17]. In this study the long extrction times t room temperture is not used, since the min objective ws to simulte household te extrction in wy tht would llow to obtin the polyphenolic compounds in green te using short time period. Totl phenolic content (TPC) of green te re presented en tble 2, the highest content ws Orientl (14.63 ± 0.53 mg gllid cid equivlent/g smple) nd the lowest ws Jibel (2.53 ± 0.25 mg gllid cid equivlent/g smple). In generl the four brnds showed decresing behviour Orientl> Lipton Hindú > Jibel. An nlysis of vrince ANOVA (tukey s multiple comprison Test, significnt p< 0.05), identified significnt differences between Orientl, Lipton nd Jibel. (, b, c) nd no significnt differences were found between Lipton nd Hindú (b). E. Venditti nd T. Bcchetti compred the totl phenol content levels in hot nd cold tes using Folin Cioclteu s regent. They found tht TPC is lwys higher in hot tes thn in cold tes for green te extrcts. The exception is with white te, where TPC is significntly higher in the cold infusion thn in the hot one. In ddition, TPC, in white te prepred with cold wter steeping, is significntly higher thn in ll other tes prepred in the sme wy [6]. This indictes tht these extrcts nlysed for te is likely to be

5 PhOL getting much smller quntities due to the extrction method. Although this only pplies to green te. As one possible reson why in our study lower content ws obtined due to temperture conditions nd extrction time. The differences found between brnds my be due to mnufcturing process used by ech industry, prticle size nd other compounds s if Jibel who contins hibiscus nd lemon peel. Totl Flvonoid content (TFC) of green te re presented en tble 2, the highest content ws Orientl (7.08 ± 0.38 mg ctechin equivlent/g smple) nd Hindú (6.71 ± 0.36 mg ctechin equivlent/g smple), while the lowest ws Jibel (2.67 ± 0.20 mg ctechin equivlent/g smple). In generl the four brnds showed decresing behviour Orientl Hindú > Lipton> Jibel. An nlysis of vrince ANOVA (tukey s Multiple Comprison Test, significnt p< 0.05), identified significnt differences between Orientl, Lipton nd Jibel. (, b, c) nd no significnt differences were found between Orientl nd Hindú(). See figure 2. Luximon-Rmm [18] showed contents from 15 to 26 mg/g for totl flvonoids in nine commercilly blck te, considering extrctions were performed t boiling temperture. With these dt reported nd compred to ours extrction continues to observe lower content of flvonoids for green te extrcts t room temperture. The ntioxidnt ctivity of the te infusions ws ssessed using two independent ssys, DPPH nd ORAC. The DPPH method is bsed on the reduction of the reltively stble rdicl 1,1-diphenyl-2-picrylhydrzyl, to the formtion of non rdicl form in the presence of hydrogen donting ntioxidnt. The te smples showed ntioxidnt ctivity by the reduction of purple colored DPPH to the yellow colored diphenylpicrylhydrzine derivtives t wvelength of 517 nm [19]. The ORAC ssy originlly developed by Glzer nd Ghiselli [20]. The ORAC ssy offers severl dvntges over DPPH ssy. It uses peroxyl rdicls tht re better models of ntioxidnt rections with oxidizing lipids nd rective oxygen species (ROS) in foods nd in vivo, nd it provides continuous genertion of rdicls on relistic time scle (more like ctul rections in situ).the completely hydrogen tom trnsfer rdicl quenching mechnism presents contrst nd comprison to electron trnsfer in DPPH ssys. The ssy cn be dpted to detect both hydrophilic nd hydrophobic ntioxidnts generlly or specificlly by ltering the rdicl source, solvent, nd trget molecule, nd hs been routinely utomted [21]. The ntioxidnt ctivity of green te re presented in tble 2, the highest content ws Orientl (41.29 ± 0.86 mg Trolox equivlent/g smple) nd Hindú (38.64 ± 1.72 mg Trolox equivlent/g smple) determined by DPPH ssy, while the lowest ws Jibel (22.36 ± 0.98 mg Trolox equivlent/g smple). In generl the four brnds showed decresing behviour Orientl Hindú>Lipton>Jibel. An nlysis of vrince ANOVA (tukey s Multiple Comprison Test, significnt p< 0.05), identified significnt differences between Orientl, Lipton nd Jibel. (, b, c) nd no significnt differences were found between Orientl nd Hindú (). See figure 3. The results obtined by ORAC llowed to see tht the highest ntioxidnt ctivity ws to Orientl te (46.25 ± 2.05 mg Trolox equivlent/g smple) nd the lowest ws Jibel (22.95 ± 1.31 mg Trolox equivlent/g smple), nd their behviour of ll smples ws Orientl >Hindú Lipton > Jibel. Antioxidnt ctivity of ll smples showed lmost the sme behviour ssessed by two methods. According to the results obtined from the nlysis performed, Jibel te brnd showed the sme results for both ssys (TPC nd TFC). Perhps t this extrction conditions ll the phenols presented in the solution re flvonoids. See tble 2. Jibel showed the lowest levels of phenolic compounds, flvonoids nd ntioxidnt ctivity, possibly due to the ddition of hibiscus nd lemon peel tht could msk other compounds [22]. Therefore from the bove results, one cn deduce tht ntioxidnt ctivity in the vrious te brnds tested is correlted with their totl flvonoid content. For DPPH ssy the correltion is given by r 2 = nd for ORAC ssy the correltion is given by r 2 = In ccordnce with the liner correltions is possible to hve some fctors tht could ese the form to get results bout ntioxidnt ctivity hving previously nlyzed the totl flvonoid content. Correltion between DPPH nd TFC hs the eqution below: TFC = A. A 0.24 Correltion between ORAC nd TFC hs the eqution below:

6 PhOL TFC = A. A 0.19 The coefficients TFC re relted to the totl flvonoid content expressed in mg gllic cid equivlent/g smple nd A.A to ntioxidnt ctivity expressed in mg Trolox equivlent/g smple. To dte, there pper to be no thorough studies on how ntioxidnt ctivity of tes my be ffected by hot or cold wter steeping nd how this my be relted to their polyphenol content. The results obtined contribute to gining further knowledge on how the potentil helth benefits of this populr beverge my be mximized by the different methods of preprtion. Acknowledgments The uthors would like to thnk the Universidd Tecnológic de Pereir for finncil support. References [1] A. Büyükblci nd S. N. El, Determintion of In Vitro Antidibetic Effects, Antioxidnt Activities nd Phenol Contents of Some Herbl Tes, Plnt Foods Hum. Nutr., vol. 63, no. 1, pp , [2] D. L. McKy nd J. B. Blumberg, The Role of Te in Humn Helth: An Updte, J. Am. Coll. Nutr., vol. 21, no. 1, pp. 1 13, Feb [3] P. C.. Hollmn nd M.. Ktn, Dietry Flvonoids: Intke, Helth Effects nd Biovilbility, Food Chem. Toxicol., vol. 37, no. 9 10, pp , Sep [4] A.-M. Mne, B. S. Vsile, nd A. Meghe, Antioxidnt nd ntimicrobil ctivities of green te extrct loded into nnostructured lipid crriers, Comptes Rendus Chim., vol. 17, no. 4, pp , [5] A. Rshidinejd, E. J. Birch, nd D. W. Everett, Antioxidnt ctivity nd recovery of green te ctechins in full-ft cheese following gstrointestinl simulted digestion, J. Food Compos. Anl., vol. 48, pp , [6] E. Venditti, T. Bcchetti, L. Tino, P. Crloni, L. Greci, nd E. Dmini, Hot vs. cold wter steeping of different tes: Do they ffect ntioxidnt ctivity?, Food Chem., vol. 119, no. 4, pp , [7] D. Komes, D. Horžić, A. Belščk, K. K. Gnić, nd I. Vulić, Green te preprtion nd its influence on the content of bioctive compounds, Food Res. Int., vol. 43, no. 1, pp , [8] C. Astill, M. R. Birch, C. Dcombe, P. G. Humphrey, nd P. T. Mrtin, Fctors Affecting the Cffeine nd Polyphenol Contents of Blck nd Green Te Infusions, J. Agric. Food Chem., vol. 49, no. 11, pp , Nov [9] V. L. Singleton, R. Orthofer, nd R. M. Lmuel- Rventós, Oxidnts nd Antioxidnts Prt A, vol Elsevier, [10] D.-O. Kim, K. W. Lee, H. J. Lee, nd C. Y. Lee, Vitmin C Equivlent Antioxidnt Cpcity (VCEAC) of Phenolic Phytochemicls, J. Agric. Food Chem., vol. 50, no. 13, pp , Jun [11] J. Zhishen, T. Mengcheng, nd W. Jinming, The determintion of flvonoid contents in mulberry nd their scvenging effects on superoxide rdicls, Food Chem., vol. 64, no. 4, pp , Mr [12] O. P. Shrm nd T. K. Bht, DPPH ntioxidnt ssy revisited, Food Chem., vol. 113, no. 4, pp , Apr [13] K. M. Gillespie, J. M. Che, nd E. A. Ainsworth, Rpid mesurement of totl ntioxidnt cpcity in plnts, Nt. Protoc., vol. 2, no. 4, pp , Apr

7 PhOL [14] F. Shhidi, Antioxidnt fctors in plnt foods nd selected oilseeds, BioFctors, vol. 13, no. 1 4, pp , Jn [15] X. Pn, G. Niu, nd H. Liu, Microwve-ssisted extrction of te polyphenols nd te cffeine from green te leves, Chem. Eng. Process., vol. 42, no. 2, pp , [16] E. Dmini, T. Bcchetti, L. Pdell, L. Tino, nd P. Crloni, Antioxidnt ctivity of different white tes: Comprison of hot nd cold te infusions, J. Food Compos. Anl., vol. 33, no. 1, pp , [17] T. Bhorun, V. Neergheen-Bhujun, N. A. Toolsee, J. Somnh, A. Luximon-Rmm, nd O. I. Aruom, Bioctive Phytophenolics nd Antioxidnt Functions of Aqueous nd Orgnic Te Extrcts. Elsevier, [18] A. Luximon-Rmm, T. Bhorun, A. Crozier, V. Zbrsky, K. P. Dtl, D. T. Dexter, nd O. I. Aruom, Chrcteriztion of the ntioxidnt functions of flvonoids nd pronthocynidins in Muritin blck tes, Food Res. Int., vol. 38, no. 4, pp , [19] Y. Zhng, Q. Li, H. Xing, X. Lu, L. Zho, K. Qu, nd K. Bi, Evlution of ntioxidnt ctivity of ten compounds in different te smples by mens of n on-line HPLC-DPPH ssy, Food Res. Int., vol. 53, no. 2, pp , [20] A. N. Glzer, Oxygen Rdicls in Biologicl Systems Prt B: Oxygen Rdicls nd Antioxidnts, vol Elsevier, [21] K. M. Schich, X. Tin, nd J. Xie, Reprint of Hurdles nd pitflls in mesuring ntioxidnt efficcy: A criticl evlution of ABTS, DPPH, nd ORAC ssys, J. Funct. Foods, vol. 18, pp , [22] L. S. Rmirez, L. F. Ospin, nd A. Ortiz, Evlution of the Antioxidnt Cpcity nd Chrcteriztion of Phenolic Compounds Obtined From Te ( Cmelli Sinensis ) for Products of Different Brnds Sold in Colombi., vol. 3, pp , 2015.

8 m g A c id g llic E q / g s m p le T e m p e r tu r e C PhOL O R IE N T A L 1 O R IE N T A L 2 O R IE N T A L 3 L IP T O N 1 L IP T O N 2 L IP T O N 3 H IN D Ú 1 H IN D Ú 2 H IN D Ú 3 J A IB E L 1 J A IB E L 2 J A IB E L E x tr c tio n d y s Figure 1. Temperture Control b b c 0 O R IEN T A L L IP T O N H IN D Ú J A IB EL Figure 2. Totl phenol content (TPC).

9 m g T R O L O X E q / g s m p le m g c te c h in E q / g s m p le PhOL 8 6 b 4 c 2 0 O R IE N T AL L IP T O N H IN D Ú J AIB E L Figure 3. Totl flvonoid content (TFC) b 3 0 c O R IE N T AL L IP T O N H IN D Ú J AIB E L Figure 4. DPPH ssy.

10 mg Ctechin Eq/g smple m g T R O L O X E q / g s m p le PhOL b c O R IE N T AL L IP T O N H IN D Ú J AIB E L Figure 5. ORAC ssy. 8,00 7,00 6,00 y = 0,2396x - 2,7637 R² = 0,9911 5,00 4,00 3,00 2,00 1,00 0,00 0,00 10,00 20,00 30,00 40,00 50,00 mg Trolox Eq/g smple Figure 6. Correltion DPPH Vs TFC.

11 mg Ctechin Eq/g smple PhOL 8,00 7,00 6,00 5,00 4,00 3,00 2,00 1,00 y = 0,1929x - 1,8105 R² = 0,9968 0,00 0,00 10,00 20,00 30,00 40,00 50,00 mg Trolox Eq/g smple Figure 7. Correltion ORAC Vs TFC. Tble 1. Weight smples. SAMPLE DAY WEIGHT (g) ORIENTAL LIPTON HINDÚ JAIBEL Tble 2. Summry, TPC, TFC, DPPH nd ORAC results of green te t room temperture. Totl phenol content Totl flvonoid content (TFC) (TPC) DPPH ssy ORAC ssy Smple mg GAE / g smple mg /g smple eq CAT mg Trolox eq/ g smple mg Trolox eq/ g smple ORIENTAL ± ± ± ± 2.05 LIPTON ± ± ± ± 1.90 HINDÚ 9.89 ± ± ± ± 1.40 JAIBEL 2.53 ± ± ± ± 1.31