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6 protocols using catechin gallate

1

Quantitative Analysis of Flavonoids

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LC-MS-grade methanol was purchased from Merck (Darmstadt, Germany). Formic acid was purchased from Fisher Scientific (LC-MS grade, Thermo Scientific, Carlsbad, CA, USA), and ammonium formate was obtained from Macklin (HPLC grade, purity: ≥99%). Sakuranetin was purchased from Sigma-Aldrich (Sigma-Aldrich, Inc., St. Louis, MO, USA, CAS number: 2957-21-3); quercitrin (CAS number: 522-12-3), myricitrin (CAS number: 17912-87-7), kaempferol (CAS number: 520-18-3), luteolin (CAS number: 491-70-3), rutin (CAS number: 153-18-4), hyperoside (CAS number: 482-36-0), kaempferol-3-O-rutinoside (CAS number: 17650-84-9), catechin (CAS number: 225937-10-0), and catechin gallate (CAS number: 130405-40-2) were purchased from Shanghai Yuanye Bio-Technology Co., Ltd. (Shanghai, China). Ultrapure water was purified using a Milli-Q system (18 MΩ*cm, Millipore, Burlington, MA, USA).
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2

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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3

Qingzhuan Tea's Bioactive Compounds

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Tea samples (Qingzhuan tea) were obtained from Huanghelou Tea Factory (Hubei, China); α-glucosidase (EC 3.2.1.20 Saccharomyces cerevisiae, G0660) and p-nitrophenyl-α-D-glucopyranoside (pNPG, N1377) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Catechins standards of (−)-epicatechin (EC), (+)-catechin (C), (−)-epigallocatechin (EGC), (−)-gallocatechin (GC), (−)-epicatechin gallate (ECG), (−)-catechin gallate (CG), (−)-epigallocatechin gallate (EGCG) and (−)-gallocatechin gallate (GCG) and gallic acid (GA) were purchased from Shanghai Yuanye Biological Technology Company Limited (Shanghai, China). Acarbose was acquired from the National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China). All other reagents and solvents were purchased from China National Pharmaceutical Group Corporation (Beijing, China) and were of analytical grade. Purified water (18.2 MΩ) was prepared using a Millipore Mill-Q Ultrapure Water System (Billerica, MA, USA).
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4

Comprehensive Tea Phytochemical Analysis

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Black tea, green tea and jasmine tea were from Eastsign Foods (Quzhou) Co., Ltd. (Quzhou, China). Purified water was from Hangzhou Wahaha Beverage Co., Ltd. (Hangzhou, Zhejiang, China), which had a total dissolved solids of 1.768 mg/L and a pH of 6.74.
Gallic acid (GA), gallocatechin (GC), epigallocatechin (EGC), catechin (C), epicatechin (EC), gallocatechin gallate (GCG), epigallocatechin gallate (EGCG), catechin gallate (CG), epicatechin gallate (ECG), theaflavin (TF), theaflavin-3-gallate (TF-3-G), theaflavin-3-gallate (TF-3-G) and theaflavin-3,3-digallate (TFDG) were from Shanghai Yuanye Biotechnology Co., Ltd. (China) with purity ≥95%. N-Alkanes for linear retention index (RI) determination and internal standard solution (decanoic acid ethyl ester) were from TCI (Shanghai) Development Co., Ltd. (Shanghai, China). Ninhydrin, stannous chloride, sodium carbonate, disodium hydrogen phosphate, potassium dihydrogen phosphate and disodium EDTA were all analytical grade from local suppliers. Acetic acid and acetonitrile were chromatographic grade, from local suppliers.
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5

Procurement of Flavonoid and Amino Acid Standards

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The following chemicals were purchased from Shanghai Yuanye Bio-Technology Co., Ltd. (Shanghai, China): catechin (C), epicatechin (EC), epigallocatechin (EGC), gallocatechin (GC), gallocatechin gallate (GCG), epigallocatechin gallate (EGCG), epicatechin gallate (ECG), catechin gallate (CG), theaflavin, theaflavin-3-gallate, theaflavin-3′-gallate, and theaflavin-3,3′-di-O-gallate. Flavonoids and glycosides standards were purchased from Shanghai Macklin Biochemical Technology Co., Ltd. (Shanghai, China), including rutin, quercetin, vitexin, myricetin, myricetin 3-O-galatoside, kaempferol-3-O-glucoside quercetin-3-O-glucoside, kaempferol-3-O-rutinoside, quercetin-3-O-glucosylrutinoside, proanthocyanidin B1, proanthocyanidin B2, proanthocyanidin B3, proanthocyanidin B4, β-glucogallin, and quinic acid. We also purchased a mixed amino acids standard from Sykam GmbH (Munich, Germany).
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6

Longjing 43 Tea Polyphenol Analysis

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Fresh leaves of Longjing 43 tea plants were plucked according to the standard method of one bud and four leaves, and then immediately stored frozen at −20 °C. Edible defoamer for bean products was from a food additives company in Hebei Province, China. Green tea pieces (Longjing) were from Hangzhou Longguan Industrial Co., Ltd. (China). Tannase (5000 U/g) was from Heilongjiang Weinuoen Biotechnology Co., Ltd. (China). Gallic acid (GA), gallocatechin (GC), epigallocatechin (EGC), catechin (C), epicatechin (EC), gallocatechin gallate (GCG), epigallocatechin gallate (EGCG), catechin gallate (CG), epicatechin gallate (ECG), TF, theaflavin-3-gallate (TF-3-G), theaflavin-3′-gallate (TF-3′-G) and theaflavin-3–3′-digallate (TFDG) were from Shanghai YuanYe Biotechnology Co., Ltd. (China) with purity of ≥ 95%. Citric acid monohydrate, disodium hydrogen phosphate (analytical grade), acetonitrile, acetic acid, water (chromatography grade) and other chemicals were from local suppliers. Purified water was from Hangzhou Wahaha Beverage Co., Ltd. (China).
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