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4 protocols using 3 hydroxyphenylpropionic acid

1

Litchi Fruit Pericarp Extraction Protocol

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Fruit of litchi (Litchi chinensis Baila) was obtained from Guangzhou and arrived in the laboratory within 24 h of harvest. Fruits were peeled and the pericarp was quickly stored at −18°C until use.
β-glucuronidase Type H-5, (-)-epicatechin, vanillic acid, caffeic acid, m-coumaric acid, p-coumaric acid, 3-hydroxyphenylacetic acid, 3-hydroxyphenylpropionic acid, 4-hydroxyphenylpropionic acid, shikimic acid, xanthine oxidase, xanthine, and uric acid were obtained from Sigma (St. Louis, MO, USA). The β-glucuronidase demonstrated both β-glucuronidase (600 U/mg solid) and sulfatase (50 U/mg solid) activity. Grape seed procyanidin was purchased from Jianfeng Natural Product R&D Co., Ltd. (Tianjin, China). Acetonitrile and methanol (HPLC grade) were purchased from Fisher Scientific (Waltham, Massachusetts, USA). All the other chemicals and reagents were of analytical grade, unless specified.
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2

Quantification of Polyphenol Standards

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Commercial standards of cyanidin-3-O-glucoside chloride, delphindin-3-O-glucoside chloride, malvidin-3-O-glucoside chloride, gallic acid, caffeic acid, ferulic acid, ethyl gallate, taxiFolin, chlorogenic acid, hippuric acid, 3-hydroxyhippuric acid, 4-hydroxybenzaldehyde, isovanillin, p-anisic acid, 4-hydroxyphenylacetic acid, 3-hydroxyphenylpropionic acid, 3-methoxyphenylacetic acid, isovanillic acid, homovanillic acid, 3-hydroxy-4-methoxyphenylpropionic acid, syringic acid, quercetin, myricetin, chlorogenic acid, quercetin-3-O-glucuronide, protocatechuic acid, p-coumaric acid, catechin, epicatechin, 4-methoxyquercetin, and quercetin-3-O-glucoside as well as sodium carbonate and Folin and Ciocalteu’s reagent (2N) were purchased from Sigma-Aldrich (St. Louis, MO, USA). caffeic acid glucuronide was supplied by Synthose (Concord, Ontario, Canada). LC-MS grade solvents, including methanol, water, ACN, and formic acid as well as trace metal grade concentrated nitric acid were purchased from Thermo Fisher Scientific (Waltham, MA, USA). 45Ca was purchased from PerkinElmer (Waltham, MA, USA). EcoLite (+) scintillation cocktail was purchased from MP Biomedicals (Santa Ana, CA, USA).
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3

Quantitative Analysis of Blueberry Polyphenols

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Commercial standards of cyanidin-3-O-glucoside chloride, malvidin-3-O-glucoside chloride, peonidin-3-O-glucoside chloride, petunidin-3-O-glucoside chloride, gallic acid, caffeic acid, ferulic acid, ethyl gallate, taxifolin, chlorogenic acid, 4-hydroxybenzoic acid, 3-hydroxybenzaldehyde, hippuric acid, 3-hydroxyhippuric acid, isovanillin, 4-hydroxyphenylacetic acid, 3-hydroxyphenylpropionic acid, 3-methoxyphenylacetic acid, vanillic acid, homovanillic acid, syringic acid, quercetin, myricetin, quercetin-3-O-glucuronide, 4-methoxyquerceting, p-coumaric acid, epicatechin, kaempferol, and quercetin-3-O-rutinoside as well as sodium carbonate and Folin and Ciocalteu’s reagent (2N) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Extraction and LC-MS grade solvents, including methanol, water, acetonitrile, hexanes, and formic acid were purchased from Thermo Fisher Scientific (Waltham, MA, USA). VitaBlue Pure American Blueberry Extract was obtained from FutureCeuticals (Momence, IL, USA); lyophilized composite wild blueberry (Vaccinium angustifolium) powder was obtained from the Wild Blueberry Association of North America (Old Towne, ME, USA).
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4

Profiling Phytochemicals in Lotus Seedpods

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Mature lotus seedpods of Nelumbo nucifera Gaertn. (cultivar: Number 2 Wuhan plant) were obtained from Honghu Lantian (Hubei, China) in late July, 2012, and identified by Professor Xueming Ni from the Department of Botany, Wuhan Plant Institute of the Chinese Academy of Science. (+)-Catechin, ferulic acid, caffeic acid, syringic acid, m-coumaric acid, 3-hydroxybenzoic acid, 3-hydroxyphenylacetic acid and 3-hydroxyphenylpropionic acid were purchased from Sigma Chemical Co. (St. Louis, MO, USA). The β-glucuronidase demonstrated both β-glucuronidase (600 U/mg solid) and sulfatase (50 U/mg solid) activity. Methanol and formic acid (HPLC grade) were purchased from Fisher Scientific (Massachusetts, USA). Phosphate buffer saline (PBS, pH 7.4), bovine serum albumin (BSA) and d-glucose were purchased from Sinopharm Chemical Reagent (Shanghai, China). All other chemicals were of analytical grade.
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