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1 RSC Advanes This is an Aepted Manusript, whih has een through the Royal Soiety of Chemistry peer review proess and has een aepted for puliation. Aepted Manusripts are pulished online shortly after aeptane, efore tehnial editing, formatting and proof reading. Using this free servie, authors an make their results availale to the ommunity, in itale form, efore we pulish the edited artile. This Aepted Manusript will e replaed y the edited, formatted and paginated artile as soon as this is availale. You an find more information aout Aepted Manusripts in the Information for Authors. Please note that tehnial editing may introdue minor hanges to the text and/or graphis, whih may alter ontent. The journal s standard Terms & Conditions and the Ethial guidelines still apply. In no event shall the Royal Soiety of Chemistry e held responsile for any errors or omissions in this Aepted Manusript or any onsequenes arising from the use of any information it ontains.

2 Page 1 of 7 Please RSC do not Advanes adjust margins RSC Advanes Reeived 00th January 20xx, Aepted 00th January 20xx DOI: /x0xx00000x Introdution Staility, antioxidant ativity and in vitro ile aids-inding of green, lak and dark tea polyphenols during simulated in vitro gastrointestinal digestion Zhengmei Wu, a Jianwen Teng, a Li Huang, a Ning Xia, a and Baoyao Wei, a, * The staility of phenoli ompounds and antioxidant ativity, as well as the ile aids-inding ativity of green, lak, raw liuao and aged liuao tea during the in vitro gastrointestinal digestion were evaluated. After the in vitro gastrointestinal digestion, the total phenoli ontent of green tea remarkly dereased. However, it inreased in the fermented tea of lak, raw liuao and aged liuao. Meanwhile, the total atehin reovery of green tea was 52.79%, ut the fermented tea of lak, raw liuao and aged liuao were %, 92.73%, and %, respetively. After gastrointestinal digestion, the ABTS ation radial savenging apaity of lak tea and aged liuao tea inreased, and the post-fermented liuao tea inreased more signifiantly. In vitro ile aid inding y tea extrats of green, lak, raw liuao and aged liuao tea inding of % ile aids. Therefore, teas have potential for hyperlipidemi prevention assoiated with ardiovasular disease. Reently, tea produts have een oserved an inreasing interest in human life. The teas an e lassified as unfermented tea (green tea), semi-fermented (oolong tea), fermented (lak tea), and post-fermented (pu-erh tea, liuao tea) on the asis of the prodution methods. In the past deades, people found that tea produts were effetive for the prevention of various illnesses 1-3.It had een reported that these effets were attriuted to the polyphenol ompounds in tea 4. In general, the teas are digested in the gastrointestinal trat. The polyphenols ompounds are released during the digestion and are asored in the intestinal trat to ahieve this speifi effet 5. Tenore et al found that the polyphenol ompounds an e dereased y average of 44.4% during the digestion, as well as 91.8% of native atehin in tea 6. Bile aid iosynthesis in the intestinal trat plays an important role in maintaining holesterol homeostasis. Potential holesterol-lowering and aner prevention aility of food frations ould e predited y evaluating the aility of inding ile aid in vitro. Colestyramine is one kind of antiholesteremi agent, and is usually employed as the positive ile aids inding ontrol in the previous studies 7-9. Both in vitro and in vivo researhes indiate that there is a positive orrelation etween the ile aid inding aility and holestyramine dosage 10,11. Binding of ile aid and enhaning the ile aid ontent in faees have een assumed to e the possiility holesterol-lowering mehanism of food frations 7-9. Gong et al found that the ontent of holesterol and ile aid in the faees of hyperlipidemia mie ould e inreased y times y the adding of pu-erh tea thearownin. It indiated that the thearownin an improve the transformation and disharge of the holesterol in food 12. Reent researhes demonstrated that phenoli sustanes in tea have the aility of inding ile aid. Ngamukote et al indiated that the ontent of ile aid ould e redued y the inding of galli aid, atehin and epiatehin with tauroholi aid, glyodeoxyholi aid hydrate and turoursodesoxyholi aid, whih thus redued the soluility of holesterol 13. These studies indiate that polyphenols suh as atehin, epiatehin, theaflavin and thearownin are the main onstituents affeting the onentration of the ile aid. Thus it an e seen that the polyphenols ompounds have the potential to ind with the ile aid, whih will onsequently inhiit the asorption of holesterol and inrease the exretion of holesterol and ile aid. In our previous studies 14,15, we found that the extrats of liuao tea have the effet of reduing lipid and antioagulation of the hyperlipidemi mie, and the major antihyperlipidemia individual atehin were finally identified. Our present study foused on the inding effet etween the major ative ingredients in tea (phenoli ompounds) and the ile aids. Changes of the phenoli ompounds and antioxidant ativity were investigated efore and after in vitro digestion. The orrelations etween the individual atehin and the ile aid inding aility were determined. All our studies showed the potential antihyperlipidemia aility of teas. RSC Advanes Aepted Manusript This journal is The Royal Soiety of Chemistry 20xx J. Name., 2013, 00, 1-3 1

3 Please RSC do not Advanes adjust margins Page 2 of 7 Experimental Chemials All reagents and hemials used were either HPLC grade or analytial grade. The water was prepared using a ompat ultrapure water system efore use. Standards were: GA: (+)- galli aid was purhased from Sigma-Aldrih (St. Louis, MO, USA). GC: ( )-galloatehin; EC: ( )-epiatehin; EGCG: ( )- epigalloatehingallate; GCG: ( )- galloatehin gallate; ECG: ( )-epiatehingallate were otained from Institute of Beijing YingzeNaxin hemial tehnology (Beijing, China). Aetonitrile (HPLC grade) was ought from Merk Speialities Private Limited (Indian). Chemials and reagents used to simulate the gastrointestinal digestion, and ile aid analysis, were: pepsin, panreatin, glyoholi aid, glyohenoholi aid, glyodeoxyholi aid, taurohenoholi aid, taurodeoxyholi aid and holestyramine, were purhased from Sigma-Aldrih (St. Louis, MO, USA). The ile aids analysis ommerial kits (TBA testing kit, Shanghai Juhuang Bioteh. Co. Ltd., (Shanghai, China). Folin Cioalteu s phenol reagent was ought from Coolaer siene & tehnology (Beijing, China). L-Asori aid (VC) was ought from Tianjing Bodi hemial industry CO., Ltd (Tianjing, China). 2,2,-azinois-(3- ethylenzothiazoline-6-sulphoni aid) diammonium salt (ABTS) was ought from Sigma Aldrih (St. Louis, MO). 1,1-Diphenyl- 2-pirylhydrazyl (DPPH) was ought from Tokyo Chemial Industry (St. Portland, Japan). Preparation of extrats and lak tea samples were otained from loal markets in Nanning City of China as individual tea ags, raw liuao tea (Slightly fermented) and 3 years aged liuao (post-fermented) tea were otained from China Tea CO., Ltd (Wuzhou, China). All teas were ground to powder. A total of g powdered sample was mixed with oiled water (50 ml) and inuated at for 5 min. After ooling, eah sample extrat was entrifuged, and supernatant was removed into a volumetri flask (50mL). An additional of deionized water was used to rinse out the entrifuge tue, whih was vortexed and entrifuged as efore. Supernatant was omined with the previous supernatant and dilute with deionized water to 50mL. The solution were frozen at -20 for further analysis. Content of total polyphenols and atehins in tea The total phenoli ontent was determined aording to the International Standards Organisation (ISO) ISO E, y the Folin Cioalteu reagent. Briefly, 1 ml of sample extrat was pipetted into a volumetri flask (100 ml) and mixed with distilled water to the mark. 1mL of the diluted sample was mixed with 5 ml of 10% Folin Cioalteu s phenol reagent. After 3-8 min, 4 ml of 7.5% sodium aronate solution was added to the reation mixture, then stand for 1 h efore spetrometri analysis. The standard urves of various onentrations of galli aid were used for quantifiation, and the results were expressed as mg of galli aid per ml of sample extrats. Analysis of individual atehins ontent y HPLC, and was performed aording to the ISO E proedure. 1 ml of the sample extrat was diluted to 10 ml with stailizing solution (10% v/v aetonitrile with 500 μg/ml EDTA and asori aid). Polyphenols were purified from the extrats after digestion y solid phase extration (SPE) using ProElut C18 artridges (Dikma Tehnologies, China). The purified solution was filtered through a 0.45 um nylon memrane filter and put into vials. An aliquot of 10 μl of the solution was injeted into the HPLC system y an auto-sampler. The system, omprised of a Waters e2695 Separations Module, a Waters 2998 Photodiode Array Detetor, with photodiode array detetion at 278 nm, using a ZORBAX Elipse Plus C18 olumn (250 mm 4.6 mm i.d., 5μm) (Agilent Tehnologies, USA). Column temperature was set at 35. Moile phase A, 9 % (volume fration) aetonitrile, 2 % (volume fration) aeti aid with 20 µg/ml EDTA. Moile phase B, 80 % (volume fration) aetonitrile, 2 % (volume fration) aeti aid with 20 µg/ml EDTA. The flow rate of the moile phase was 1 ml/min. The gradient of moile phase A as follows: 0-10 min, 100%, min, from 100% -68% and kept at this omposition for 10 min. Then reset to 100% A and allowed to equilirate 10min efore the next injetion. Catehin and galli aid were identified y omparison of their retention times and spetra to those of the standards. Quantifiation of atehin and galli aid was arried out y integrating the peak areas and using aliration urves. Results were expressed as mirogram of eah phenoli ompound per 1 ml of sample. Binding of ile aid In vitro digestion proedure The assay was performed aording to a modifiation y Kahlon & Smith 9 as follows. The ile aid mixture (36 mmol/l) ontained glyoholi aid (9 mmol/l), glyohenoholi aid (9 mmol/l), glyodeoxyholi aid (9 mmol/l), taurohenoholi aid (4.5 mmol/l) and taurodeoxyholi aid (4.5 mmol/l) in ph 6.8 phosphate uffer. This stok solution of was stored in the -20 freezer and diluted to the working solution (0.72 μmol/ml) efore eah assay. Three repliates of 1mL of green tea, lak tea, raw liuao tea and aged liuao tea extrat, holestyramine 50 mg were tested. Four sustrate lanks, one positive lank and three treatment repliates were weighed into 50mL srew-apped tues. Samples were digested in 1 ml 0.01 mol/l HCl for 2h in a 37 shaker ath. After simulating gastri digestion, the sample ph was adjusted to with 0.2 mol/l NaOH. Add 4 ml of ile aid mixture (0.72 μmol/ml) in a phosphate uffer, ph 6.8. A phosphate uffer (4 ml, ph 6.8) was added to the sustrate lanks. Following y 5 ml of porine panreatin (8 USP, 10 mg/ml, in a Mphosphate uffer, ph 6.8) was added, tues were inuated for 2 h in a 37 shaker ath. Mixtures were entrifuged at 6000rpm for 20 min at 25. Supernatant was removed into a seond set of laeled tues. Aliquots of pooled supernatant were frozen at -20 for ile aids analysis. Bile aid analysis The ile aids were measured olorimetrially with ommerial kits. The ile aid inding ativity was alulated as: (%) = where Amixture is the ile aid onentration in the positive lank and Asupernatant is the ile aid onentration in the supernatant. Antioxidant ativity Free radial (DPPH) savenging assay RSC Advanes Aepted Manusript 2 J. Name., 2012, 00, 1-3 This journal is The Royal Soiety of Chemistry 20xx

4 Page 3 of 7 Please RSC do not Advanes adjust margins Experiments were arried out as a modifiation y Xiao et al. 16. Briefly, 2.0 ml of test sample mixed with 2.0 ml of 0.2 mm DPPH that was dissolved in 100% ethanol. The mixture was shaken well and stand at room temperature in the dark. The asorane of reation mixture was reorded after 30 min at 517 nm. Vitamin C was used as a standard ompound. The standard urves were aquired y plotting the DPPH savenging of vitamin C (ranging from 1 to 25 mg/l). The results were alulated y Y (savenging ratio) = X (VC equivalents ontent) (R2=0.991). All samples were analyzed in tripliate. ABTS ation radial savenging apaity The ABTS assay was developed y the referene of Du et al mlof 7 mm ABTS+ solution was mixed with 178 μl of 140 mm potassium persulfate (K2S2O8) in the dark at room temperature for 13 h prior to use. The mixture was diluted with PBS uffer to an asorane at 0.70 ± 0.05 at 734 nm. An aliquot of 0.1 ml of sample, lank (water) and standard mixed with 3.9 ml of diluted ABTS+ to reat in the dark at room temperature, and asorane was reorded at 734 nm after 5 min. Vitamin C was used as a standard ompound. The standard urves were aquired y plotting the ABTS ation radial savenging apaity of vitamin C (ranging from 1 to 25 mg/l). The results were alulated y Y (savenging ratio) = X (VC equivalents ontent) (R2=0.9989). All samples were analyzed in tripliate. Statistial analysis All data are presented as mean ± SD for three in vitro inuations that were analyzed in tripliate. The statistial analyses were performed using SPSS Differenes of P<0.05 were onsidered signifiant. Data were sujeted to 1- way ANOVA, means ompared using Tukey s test (p<0.05). Correlations among the antioxidant apaity, the total polyphenoli ontent and the ile aids-inding apaity were estalished using regression analysis at a 95% signifiane level. The orrelations was onsidered signifiant when P<0.05. Results and disussion Changes of phenoli ompounds The phenoli ompounds are harateristi onstituents in tea, and they are also the main ative ingredient in tea. Changes of total phenoli ompounds in green, lak, raw liuao and aged liuao tea extrats during the simulated in vitro gastrointestinal digestion were shown in Fig. 1. It an e seen that there was great differene of total phenoli ompounds ontent etween different kinds of tea samples. The total phenoli ompounds ontent of green tea was higher than the other teas, and it was higher in liuao tea than that in lak tea. Wu et al showed that the liuao tea have the highest tea polyphenol ontent than the other dark teas in China 18. It indiated that the potential iologial ativity of liuao tea may e stronger than lak tea though it elongs to post-fermented tea. The ontent of the total phenoli ompounds in green tea (unfermented) was redued y 6.12% after simulated digestion. However, ontent of the total phenoli ompounds in lak tea (fermented tea), raw liuao tea (fermented tea) and aged liuao tea (post-fermented) were inreased y 10.06%, 3.11% and 26.86%, respetively. This indiated that the total phenoli ompounds ould e redued in unfermented teas and e inreased in fermented tea after the simulated in vitro gastrointestinal digestion. In addition, ontent of the total phenoli ompounds inreased with the inrease of the fermentation degree. The derease of the total phenoli ompounds may e eause of the degradation of atehins during the in vitro gastri and small intestinal digestion 19,20. However, the phenoli polymers in fermented teas will e degraded during the gastrointestinal digestion, whih thus produed more phenoli hydroxyl 21,22. Total phenoli ompounds ontent (mg/ml) a d d Blak tea Nondigestion sample Digestion sample raw Liuao tea Aged Liuao tea Fig. 1 Content of total phenoli ompounds in green, lak, raw liuao and aged liuao tea during the simulated in vitro gastrointestinal digestion. a Means etween different kinds of tea samples after in vitro gastro-intestinal digestion with the same letter in row are not signifiantly different aording to Tukey s multiple range test (P > 0.05). Catehin ontent of tea samples There was a positive orrelation relationship etween the atehin levels and tea radial savenging ativity. The high atehin levels may e responsile for the protetive effets of tea in onditions related to oxidative stress, neoplasti transformations and ardiovasular disease 23,24. Changes of atehin ontents in green, lak, raw liuao and aged liuao tea after simulated digestion were shown in Fig. 2-3 and Supplement 1. It indiated that total atehin reovery of green, lak, raw liuao and aged liuao tea were 52.79%, %, 92.73% and %, respetively, after gastri intestinal digestion. It an e seen from Figs. 3 and 4 that the total atehin ontents of green, lak, raw liuao and aged liuao tea were μg/ml, μg /ml, μg /ml and μg /ml, respetively. The ontent of individual atehin was dereased the most in green tea after gastri intestinal digestion. However, ontent of individual atehin was more stale in fermented tea and inreased with the inreasing period of fermentation. After the digestion, individual atehin ontent of GA, GC, EC, EGCG, GCG, ECG were all dereased in green tea. GA, GC, EC, ECG were dereased and EGCG and GCG were inreased in lak tea. GA, GC, EGCG were dereased and EC, GCG and ECG were inreased in raw liuao tea. Furthermore, GA, GC, EC, EGCG, GCG, ECG were all inreased in aged liuao tea. Many studies have proposed that atehin will degrade during the in vitro simulated gastro-intestinal digestion 19,20, these studies have RSC Advanes Aepted Manusript This journal is The Royal Soiety of Chemistry 20xx J. Name., 2013, 00, 1-3 3

5 Please RSC do not Advanes adjust margins Page 4 of 7 shown that the atehins in oth green and lak teas will e oviously degraded at alkaline ph. This degradation isn t proaly due to the derease of antioxidant ativity or total polyphenoli ontent, whih maye eause of the formation of dimers. Our results suggested that the atehins of fermented tea water extrats will degrade and regenerate in gastrointestinal trat. This maye attriute to the phenoli polymers that generated during the fermentation proess. The phenoli polymers will e degraded further and individual atehins and other ompounds will generate. Catehin ontents (µg/ml) Blak tea Raw Liuao tea Aged Liuao tea 0 GA GC EC EGCG GCG ECG Fig.2 Catehin ontent of tea samples. Catehin ontents are expressed as mean value (μg/ml tea extrats) ± SD (n = 3). Antioxidant ativity of tea extrats The antioxidant ativities of the tea extrats were usually expressed as Vitamin C Equivalent Antioxidant Capaity (VCEAC) in mg/ml. ABTS and DPPH are two kinds of stale radial speies that usually employed for antioxidant ativity measurements of the plant extrats 25,26. They are ommonly used independently to evaluate their effiaies. Aura et al indiated that the surviving phenoli ompounds during the gastri-intestinal digestion were likely to reah the olon. These phenoli ompounds will at as antioxidants or e iotransformed into phenoli antioxidants and e asored in the large intestine/olon 27. Our results as shown in Fig. 4 and Fig.5 indiated that the digestion produts still possessed ertain antioxidant ativities and the order was Nondigestion sample >Digestion sample, exept ABTS ation radial savenging apaity of lak tea and aged liuao tea. DPPH radial savenging ativity of green, lak, raw liuao and aged liuao tea showed a similar tendeny during the in vitro gastro-intestinal digestion as shown in Fig. 4. The DPPH radial savenging ativity of green, lak, raw liuao and aged liuao tea were dereased y 55.9%, 58.7%, 54.0% and 54.2%, respetively. It indiated that the DPPH radial savenging ativity of all tea extrats remarkly dereased after vitro gastro-intestinal digestion. This ould e due to the fermenting degrees make no differene to the DPPH savenging ativity of tea, or it may e that the oxidation resultant produts also have remarkale effet on the antioxidant ativity 28. ABTS ation radial savenging apaity of green, lak, raw liuao and aged liuao tea were shown in Fig. 5. During vitro gastro-intestinal digestion, ABTS ation radial savenging apaity of green and raw liuao tea were dereased y 10.7% and 36.45%, respetively. However, ABTS ation radial savenging apaity of lak and aged liuao tea were inreased y 22.9% and 26.0%, respetively. It ould e speulated that the tea polyphenols polymerides struture will e hanged and will e degraded during its transportation from intestine. The resultant small moleular phenoli ompounds and reakdown produts might have a higher ABTS ation radial savenging apaity, whih will diretly ontriute to the antioxidant apaity of the digestion samples. Mukai et al indiated that the inrease of the radial savenger ativity was attriuted to the deprotonation of the hydroxyl moieties present on the aromati rings of the phenoli ompounds 29. The phenoli moleules struture will e hanged during the transition from the stomah to the intestinal environment, whih is attriuted to the ionisation of the hydroxyl groups 30. Catehin ontents (µg/ml) Blak tea Raw Liuao tea Aged Liuao tea 0 GA GC EC EGCG GCG ECG Fig. 3 Gastro-intestinal ioaessiility of atehin from tea samples. Catehin ontents are expressed as mean value (ug/ml tea extrats) ± SD (n = 3). VCEAC (mg/ml) DPPH Blak tea Nondigestion sample Digestion sample a a a a Raw Liuao tea Aged Liuao tea Fig. 4 Antioxidant ativity (DPPH) of eah sample (mg/ml extrat) (mean ± SD, n = 3) after in vitro simulated gastrointestinal digestion with the referene standards V. a Means etween different kinds of tea produts after in vitro gastro-intestinal digestion with the same letter in row are not RSC Advanes Aepted Manusript 4 J. Name., 2012, 00, 1-3 This journal is The Royal Soiety of Chemistry 20xx

6 Page 5 of 7 Please RSC do not Advanes adjust margins signifiantly different aording to Tukey s multiple range test (P > 0.05). VCEAC (mg/ml) ABTS a a Blak tea a Nondigestion sample Digestion sample Raw Liuao tea Aged Liuao tea Fig. 5 Antioxidant ativity (ABTS) of eah sample (mg/ml extrat) (mean ± SD, n = 3) after in vitro simulated gastrointestinal digestion with the referene standards V. a Means etween different kinds of tea produts after in vitro gastro-intestinal digestion with the same letter in row are not signifiantly different aording to Tukey s multiple range test (P > 0.05). In vitro ile aid inding y tea extrat Cholestyramine is a ile aid sequestrant. It is positively harged non-digestile resins that ind to ile aids in the intestine to form an insolule omplex 31. In our researh, holestyramine ound 95% of the ile aids. It is higher than the previous studies, Story & Krithevsky found that 81% ile aid inding y holestyramine using 50 mg of sustrate and 50 μmol of ile aids 32. It may e that the use of the higher sustrate to ile aid ratio. In vitro ile aid inding y tea extrats on equal volume of tea extrat was shown in Tale1. Green, lak, raw liuao and aged liuao tea ound 1.59, 0.46, 0.65 and 0.90 μm of ile aid per milliliter of sample, respetively, whih was equal to 43.55, 12.54, and % of the total added BA. Assigning a ile aid inding value of 100% to holestyramine (50mg), the relative ile aid inding on 1mL for the test samples of tea extrat was green tea 45.83%, lak tea 13.20%, raw liuao tea 18.86%, and aged liuao tea 26.03%. Bile aid inding for green tea was signifiantly higher than the other three teas. Relative ile aid inding on tea extrat was green tea > aged liuao tea > raw liuao tea > lak tea. The different ile aid inding may e assoiate with the phytonutrients (antioxidants, polyphenols, hydroxyinnami aids, flavonoids, anthoyanins, flavonols, proanthoyanidins, atehins), hyrophoiity or ative inding sites of the various tea 9. On a tea extrat, ile aid inding of % for green, lak, raw liuao and aged liuao tea are very enouraging and ould indiate health promoting enefits of these teas. Health properties of the food frations were generally evaluated y testing their ile aid inding. Fat asorption, aner ausing toxi metaolites exretion and holesterol utilization to synthesize more ile aids will e redued y ile aids inding. It is supposed to e the lower holesterol and prevent aner mehanism y food frations. Previous studies reported for fresh fruits and fresh green vegetales, lueerries 7%, plums 6%, prunes 5%, strawerries 5%, herries 5%, ranerries 4%, apples 1%, rooli 5%, mustard greens 4%, and potato ultivars as raw, steamed or steamed then ooled range from 20.55% to 36.50% under similar onditions 9,33. It showed that ile aid inding apaity y tea extrats is signifiant higher than a variety of fruits and vegetales. Tale1. In vitro ile aid inding y tea extrat on equal volume of tea extrat A Bile aid inding Perentage Relative to Treatment (μmol/ml tea ile aid Cholestyramine extrats) inding (%) (50mg),% 1.59±0.25 a 43.55±6.84 a 45.83±7.20 a Blak tea 0.46± ± ±8.34 Raw Liuao tea 0.65± ± ±5.58 Aged Liuao tea 0.90± ± ±4.80 A Mean ± SEM within a olumn with different supersript letters differ signifiantly (P 0.05), n =3. Correlation analysis Correlation analysis was employed to go to show the relationship among total polyphenoli ontent, savenging ativities and ile aids-inding apaity. The orrelation analysis was measured for all samples during vitro gastrointestinal digestion and was presented in Tale 2. It indiated that the orrelation etween TP ontents and DPPH ativity was poor. This may e due to TP did not have the effet of the DPPH savenging ativity of tea 34. It meant that TP ontriute less to the DPPH antioxidant apaity of fermented teas. There was a positive orrelation relationship etween the individual phenoli ompounds and the DPPH savenging ativity. This may e due to the struture hanging of the phenoli ompounds during the in vitro gastrointestinal digestion, and the newly produed ompounds also ontriute to the DPPH Tale 2 Correlation oeffiient amongst total polyphenoli ontent, savenging ativities and ile aids-inding apaity DPPH savenging apaity ABTS ation radial savenging apaity Bile aids-inding apaity TP GA GC EC EGCG GCG ECG DPPH savenging apaity 0.802* 0.819* 0.827* ABTS ation radial savenging apaity 0.947* * 0.960* 0.923* 0.970* *Signifiant at 5% level. This journal is The Royal Soiety of Chemistry 20xx J. Name., 2013, 00, RSC Advanes Aepted Manusript

7 Please RSC do not Advanes adjust margins Page 6 of 7 antioxidant apaity 19. However, highly signifiant positive orrelations of TP, GA, GC ontents with ABTS ation radial savenging apaity were oserved. This result was similar to the earlier researh that stated that TP ontent had orrelation oeffiients (r=0.836) with ABTS 35. The other individual atehins also have positive orrelations with ABTS ation radial savenging apaity, ut not signifiant. Chang et al indiated that the phenoli ontents in plants ontriuted to antioxidant ativity proaly eause of their redox properties, where the phenoli ontents ated as reduing agents, hydrogen donors and singlet oxygen quenhers 36. There was a signifiant positive orrelation etween TP, EGCG, GCG, ECG ontents and ile aids-inding apaity. Signifiant positive orrelation etween DPPH savenging apaity and ile aids-inding apaity were also oserved. The other individual atehins also have positive orrelations with ile aids-inding apaity, ut not signifiant. Ikeda et al found that tea atehins rih in galloatehin gallate and atehin gallate were etter availaility to derease holesterol asorption than that rih in epigalloatehin gallate and epiatehin gallate 37. It an thus e seen that the esterifiale atehins have higher ile aids-inding apaity than the unesterified atehins. This was in aordane with our previous results. Conlusions The tea has a positive funtion in health promotion. Phenoli polymers ould e degraded and produe individual atehins and some other ompounds during the in vitro gastrointestinal digestion of fermented tea extrats. These individual atehins have large ontriution to ABTS ation radial savenging apaity, and the interesting unknown ompounds deserve further exploration. The tea extrats possess exellent antioxidant apaity and in vitro ile aid inding apaity, where the liuao tea show more advantage than the other fermented tea. Fermented tea like liuao tea has a wider spae in the funtional researh field, whih requires more in-depth researh in future. Aknowledgements This Projet was graiously sponsored y the Guangxi Natural Siene Foundation (2012GXNSFBA053024), and the Guangxi siene and tehnology development program ( ). Gratitude is expressed to Dr. Shuangxi Nie of Guangxi University, China, who reviewed and heked this paper arefully. Notes and referenes 1. I. Ikeda, Y. Imasato, E. Sasaki, M. Nakayama, H. Nagao, T. Takeo, F. Yayae and M. Sugano, Biohimia et Biophysia Ata (BBA)-lipids and lipid MetaoLism, 1992, 1127, J. H. Weisurger and F.-L. Chung, Food and Chemial Toxiology 2002, Q. Huang, S. Chen, H. Chen, Y. Wang, Y. Wang, D. Hohstetter and P. Xu, Food and hemial toxiology : an international journal pulished for the British Industrial Biologial Researh Assoiation, 2013, 53, J. Y. Kohei Jou, Sauro Yoshioka,HironoriMoriyama,Shuzo Murata, and H. Masao Ohishi, Mitsuhiko Miyamura, Food Researh International, 2013, 54, P. Xiao, H. Huang, J. Chen and X. Li, Journal of ethnopharmaology, 2014, 157, G. C. Tenore, P. Campiglia, D. Giannetti and E. Novellino, Food Chem, 2015, 169, R.-S. Fátima, R.-G. Guillermo, L.-M. Antonio and F.-B. Juan, Food Hydroolloids, 2015, 43, Y. Niu, Z. Xie, J. Hao, W. Yao, J. Yue and L. Yu, Food Chemistry, 2012, 132, T. Kahlon and G. Smith, Food Chemistry, 2007, 100, K. E. Sukling, G. M. Benson, B. Bond, A. Gee, A. Glen, C. Haynes and B. Jakson, Atheroslerosis, 1991, 89, T. Nakamura and Y. Matsuzawa, Nihon rinsho. Japanese journal of linial mediine, 1994, 52, J. Gong, C. Peng, T. Chen, B. Gao and H. Zhou, Journal of food siene, 2010, 75, H S. Ngamukote, K. Makynen, T. Thilaweh and S. Adisakwattana, Moleules, 2011, 16, L. Huang, J. Peng, N. Xia, J. Teng and B. Wei, Food Siene and Tehnology, 2013, 8, J. Peng, Guangxi University, P. Xiao, H. Huang, J. Chen and X. Li, J Ethnopharmaol, 2014, 157, H. Du, J. Wu, H. Li, P.-X. Zhong, Y.-J. Xu, C.-H. Li, K.-X. Ji and L.- S. Wang, Food hemistry, 2013, 141, C. Wu, Hunan agriultural university, A. P. Neilson, A. S. Hopf, B. R. Cooper, M. A. Pereira, J. A. Bomser and M. G. Ferruzzi, Journal of agriultural and food hemistry, 2007, 55, I. R. Reord and J. M. Lane, Food Chemistry, 2001, 73, J. Bouayed, L. Hoffmann and T. Bohn, Food Chem, 2011, 128, M. J. Rodriguez-Roque, M. A. Rojas-Grau, P. Elez-Martinez and O. Martin-Belloso, Journal of agriultural and food hemistry, 2013, 61, G. C. Tenore, P. Stiuso, P. Campiglia and E. Novellino, Food hemistry, 2013, 141, C. J. Dufresne and E. R. Farnworth, The Journal of nutritional iohemistry, 2001, 12, L. G. Ranilla, Y.-I. Kwon, E. Apostolidis and K. Shetty, Bioresoure Tehnology, 2010, 101, D. Malenčić, Z. Maksimović, M. Popović and J. Miladinović, Bioresoure tehnology, 2008, 99, A.-M. Aura, P. Martin-Lopez, K. A. O Leary, G. Williamson, K.- M. Oksman-Caldentey, K. Poutanen and C. Santos-Buelga, European journal of nutrition, 2005, 44, RSC Advanes Aepted Manusript 6 J. Name., 2012, 00, 1-3 This journal is The Royal Soiety of Chemistry 20xx

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