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Epicatechin

Manufactured by Yuanye Bio-Technology
Sourced in China

Epicatechin is a natural compound extracted from various plant sources. It functions as a key antioxidant molecule, playing a role in scavenging free radicals and supporting cellular health. The core function of Epicatechin is to provide antioxidant protection at the molecular level.

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28 protocols using epicatechin

1

Analytical Standards for Tea Bioactives

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Catechin (C, ≥98%), galloCatechin (GC, ≥98%), galloCatechin-3-gallate (GCG, ≥98%), epiCatechin (EC, ≥98%), epiCatechin-3-gallate (ECG, ≥98%), epigalloCatechin (EGC, ≥98%), epigalloCatechin-3-gallate (EGCG, ≥98%), caffeine (CAF, ≥98%), theaflavins (TF, ≥95%), theaflavins-3-gallate (TF3G, ≥98%), theaflavin-3′-gallate (TF3′G, ≥98%), theaflavine-3,3′-digallate (TFDG, ≥98%), quercetin-3-O-galactoside (≥98%), myricetin-3-O-galactoside (≥98%), kaempferol-3-O-rutinoside (≥98%), vitexin (≥98%), astragaloside (≥98%), quercetin (≥98%), and kaempferol (≥98%) were purchased from Shanghai yuanye Bio-Technology Co., Ltd. (Shanghai, China). l-alanine, l-arginine, l-aspartic acid, l-cystine, l-glutamic acid, glycine, l-histidine, l-isoleucine, l-leucine, l-lysine, l-methionine, l-phenylalanine, l-proline, l-serine, l-threonine, l-tyrosine, l-valine, l-asparagine, l-glutamine, theanine, γ-aminobutyric acid, and l-tryptophan were purchased from Sigma (Sigma-Aldrich Co., St. Louis, MO, USA). Anthrone reagent and ninhydrin/formic acid reducing agent were purchased from BeiJing DingGuochangSheng Biotech. Co., Ltd. (Bejing, China). Ethyl caprate (≥99%) was purchased from Shanghai Macklin Biochemical Technology Co., Ltd. (Shanghai, China), and n-alkane mixed standard C7-C40 was purchased from o2si (o2si smart solutions—an LGC Standards Company, Charleston, SC, USA).
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2

Quantification of Flavonoids and Catechins

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Xanthine oxidase powder was purchased from Yuanye Biotechnology Co. (Shanghai, China). Flavone (99.0%), 7-OH flavone (98.0%), chrysin (98.0%), biochanin A (98.0%), naringin (98.0%), and myricetin (98.0%) were bought from Alfa Aesar (Thermo Fisher Scientific, MA, USA). Formononetin (99.0%) and genistein (99.0%) were acquired from Acros Organics (Fisher Scientific, PA, USA). Baicalin (98.0%), baicalein (98.0%), wogonin (98.0%), apigenin (98.0%), luteolin (98.0%), hispidulin (98.0%), tangeretin (98.0%), nobiletin (98.0%), galangin (98.0%), kaempferide (98.0%), kaempferol (98.0%), kaempferitrin (98.0%), quercetin (98.0%), fisetin (98.0%), rutin (98.0%), daidzein (98.0%), tectorigenin (98.0%), puerarin (98.0%), dihydromyricetin (98.0%), naringenin (98.0%), catechin (C; 98.0%), (−)-gallocatechin (GC; 98.0%), epicatechin, (EC; 98.0%), (−)-epicatechin gallate (ECG; 98.0%), (−)-epigallocatechin (EGC; 98.0%), (−)-epigallocatechin gallate (EGCG; 98.0%), and (−)-gallocatechin gallate (GCG; 98.0%) were commercially purchased from Yuanye Biotechnology Co. (Shanghai, China). High-performance liquid chromatography (HPLC) grade acetonitrile was purchased from Merck KGaA (Darmstadt, Germany). Ultrapure water (18.2 MΩ cm resistivity) was obtained from an ELGA water purification system (ELGA Berkefeld, Veolia, Germany). All other reagents and solvents were of analytical grade.
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3

Bioactive Compounds Extraction and Evaluation

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The 2, 2-diphenyl-1-picrylhydrazyl (DPPH), 2, 2′-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2, 4, 6-Tris(2-pyridyl)-s-triazine (TPTZ), 2, 6-di-tert-butyl-4-methylphenol (BHT), vitamin C (Vc), were obtained from Solarbio Life sciences (Beijing, China). All chemical reagents including methanol, ethyl acetate (EtOAc), petroleum ether (PE), Dichloromethane (DCM), and n-butanol (n-BuOH) were purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). Chromatographic acetonitrile was purchased from Merck (Darmstadt, Germany). Folin-Ciocalteu, pyrogallol, and other analytical grade chemicals were purchased from Aladdin (Shanghai, China). Myricetin, dihydroMyricetin, quercetin, resveratrol, apigenin, baicalein, hesperitin, gallic acid, rutin, luteolin, epicatechin, epicatechin gallate, and other standards for HPLC were purchased from Yuanye Bio-Technology (Shanghai, China). Cellulase (3, 000 μ/g) and pectinase (40, 000 μ/g) were purchased from Xiya reagent (Shandong, China). α-Glucosidase (G5003) and acetylcholinesterase (AChE, C3389) were purchased from Sigma-Aldrich (St. Louis, USA). p-Nitrophenol nitrophenol α-D-glycopyranoside (pNPG), acarbose, galantamine, and cisplatin were purchased from Sigma-Aldrich (St. Louis, MO, USA). The HepG2, MCF-7, Raw264.7, and A549 cells were purchased from Conservation genetics CAS Kunming cell bank.
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4

Comprehensive Optimization of HPLC Protocols

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Ultrapure water was prepared using a Milli-Q purifying system (Millipore, Bedford, MA, USA). Methanol and acetonitrile of HPLC grade were purchased from Merck (Damstadt, Germany). Standard compounds of histidine (1), arginine (3), cysteine (4), asparagine (5), serine (6), lysine (7), glutamine (8), proline (9), cytidine (10), hypoxanthine (11), deoxycytidine (12), uridine (13), tyrosine (14), guanine (15), guanosine(16), inosine(17), deoxyguanosine (19), isoleucine (20), leucine (21), thymidine (23), phenylalanine (24), tryptophan (27), epicatechin (33), rutin (34), hyperoside (35), and quercitrin (37) were purchased from Shanghai Yuanye Biotechnology (Shanghai, China); glutamic acid (2), gallic acid (22) and apigenin (43) were obtained from Chinese National Institute of Control of Pharmaceutical and Biological Products (Beijing, China); fumaric acid (18), gallocatechin (25), epigallocatechin (28), cryptochlorogenic acid (31), kaempferol 3-O-rutinoside (39) and amentoflavone (42) were acquired from Chengdu Chroma Biotechnology (Chengdu, China); neochlorogenic acid (26), chlorogenic acid (29), catechin (30), caffeic acid (32), isoquercitrin (36), avicularin (38), trifolin (40), and astragalin (41) were bought from Baoji Chenguang Biotechnology Co., LTD. (Baoji, China) with the purity greater than 98% and their structures were presented in Figure S4.
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5

Authentic Standards for Phytochemical Analysis

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Kaempferol, isoquercitrin, ω-hydroxyemodin, quercetin, quercitrin, rutin, luteolin, emodin-8-O-β-d-glucoside, epicatechin, catechin, and emodin were purchased from Shanghai Yuanye Biotechnology Co., Ltd. The purities of these authentic standards were >98%. HPLC-grade acetonitrile was acquired from Honeywell (Morris, NJ, USA). Acetic acid and ammonium acetate (HPLC-grade) were obtained from Sigma-Aldrich (Steinheim, Germany). Pure distilled water from Watsons water (Hong Kong, China) was used.
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6

Preparation of Polyphenol Standard Solutions

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Chlorogenic acid (CAS: 327-97-9), neoChlorogenic acid (CAS: 906-33-2), trans-cinnamic acid (CAS: 140-10-3), ferulic acid (CAS: 1135-24-6), p-coumaric acid (CAS: 501-98-4), caffeic acid (CAS: 331-39-5), p-hydroxybenzoic acid (CAS: 99-96-7), epicatechin (CAS: 490-46-0), (+)-catechin (CAS: 154-23-4), (−)-epigallocatechin (CAS: 970-74–1), (−)-gallocatechin (CAS: 3371-27-5), phloretin (CAS: 60-82-2) and phlorizin (CAS: 60-81-1) were purchased from Shanghai Yuanye Bio-technology Co., Ltd (Shanghai, China). All phenolic chemicals were analytical standards, with HPLC purity ≥ 98%.
Stock solutions of polyphenols were obtained by dissolving each polyphenol in methanol containing 5% dimethyl sulfoxide (DMSO). Subsequently, each polyphenol was diluted with sterilized NB medium to the appropriate concentration. All solutions were prepared fresh and filter-sterilized (pore size, 0.22 μm) prior to use.
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7

Polyphenol Standards and Antioxidant Assays

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Acetonitrile (HPLC-grade) was purchased from Adamas reagent, Ltd. (Shanghai, China). All the polyphenol standards (gallic acid, proanthocyanidin B2, epicatechin, proanthocyanidin A2, syringic acid, p-hydroxybenzoic acid, poncirin, ferulic acid, rutin, phlorizin, and methyl hesperidin), purity ≥99%, were purchased from Shanghai yuanye biotechnology Co. Ltd. (Shanghai, China). Vitamin C; 1, 1-diphenyl-2-picrylhydrazyl (DPPH); 2, 2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS); 2, 3, 5-triphenyltetrazolium chloride (TPTZ); triton; tris-HCl; paraformaldehyde; and urethane were purchased from the Beijing Solaibao Technology Co. Ltd. (Beijing, China). Trizol reagent was purchased from TaKaRa (Beijing, China). Hematoxylin and eosin (H&E) kit, Oil Red O (ORO) kit, and a protein extraction kit were obtained from Beyotime Biotechnology (Beijing, China). Antibodies were purchased from Abcam (Shanghai, China).
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8

Standardization of Tea Compound Analysis

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The references of non-volatile compounds (HPLC grade ≥ 98%) were purchased from Yuanye Bio-Technology Co., Ltd. (Shanghai, China), including catechins (catechin, epicatechin, catechin gallate, epicatechin gallate, gallocatechin, epigallocatechin, gallocatechin gallate and epigallocatechin-3-gallate), alkaloids (caffeine, theobromine and theophylline), flavonol glycosides (quercetin-3-O-glucoside, quercetin-3-O-galactoside, quercetin-3-O-rutinoside, quercetin-3-O-rhamnosyl-glucoside, myricetin-3-O-glucoside, myricetin-3-O-galactoside, myricetin-3-O-rhamnoside, kaempferol-3-O-glucoside, kaempferol-3-O-rutinoside, kaempferol-3-O-rhamnosyl-glucoside) and free amino acids (glutamic acid, aspartic acid, asparaginate, serine, glutamine, histidine, tryptophan, glycine, phenylalanine, arginine, theanine, alanine, γ-aminobutyric acid, tyrosine, threonine, valine, isoleucine, leucine and lysine). Acetonitrile and methanol were HPLC grade and were purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). The ultra-pure water (>18 MΩ cm) was prepared by Milli-QTM reference system (Merck Millipore, Milford, MA, USA).
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9

Polyphenol Quantification and Characterization

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The (+)-catechin, (−)-epicatechin, (−)-epigallocatechin, and (−)-epicatechin gallate (+)-gallocatechin and (−)-epigallocatechin gallate, (purity > 98 %) were purchased from Shanghai Yuanye Biotechnology Co. ltd (Shanghai, China). Methanol, acetonitrile, acetic acid, formic acid, hydrochloric acid (HPLC grade), gallic acid, rutin, and bovine serum albumin (BSA) were purchased from Sigma-Aldrich (Shanghai, China). Phloroglucinol and ascorbic acid were purchased from Shandong Xiya Chemistry Industry ltd (Linyi, China). Sodium dodecyl sulfate (SDS), triethanolamine (TEA), and Folin-phenol were purchased from Beijing Solarbio Science & Technology Co., ltd (Beijing, China). Sodium carbonate, sodium hydroxide, aluminum chloride, sodium nitrite, and sodium acetate were purchased from Tianjin Bodi Chemical Co., ltd (Tianjin, China). The C-18 SPE column was obtained from Tianjin Chengzhou Technology Co. ltd (Tianjin, China). The deionized water was obtained from a Milli-Q system (Merck Millipore, Darmstadt, Germany).
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10

Phytochemical Analysis and Antioxidant Evaluation

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Na2CO3, Al(NO3)3, formic acid, NaNO2, HCl, ethanol, Folin–Ciocalteu phenol reagent, and Trolox were obtained from Chengdu Kelong Chemical Reagent Works (Chengdu, China). 1,1-Diphenyl-2-picrylhydrazyl (DPPH·, >99.7%), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS·+, >99.7%), and chromatographic-grade methanol and acetonitrile were purchased from American Sigma (St. Louis, MO, USA). Standard compounds including (+)-catechin, (−)-epicatechin, rutin, afzelin, astragaline, kaempferol-3-O-rutinoside, kaempferol, quercetin, isoquercitrin, pinocembrin, pinocembrin 7-O-glucoside, thonningianin A, gallic acid, and pinocembrin 7-O-(3″-O-galloy-4″,6″-hexahydroxydiphenoyl)-glucoside were purchased from Shanghai Yuanye Bio-Technology Co., Ltd. (Shanghai, China). Alcohol dehydrogenase (265 units/mg) from saccharomyces cerevisiae was purchased from Sigma-Aldrich, Co. (St. Louis, MO, USA).
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