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2 protocols using formononetin

1

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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2

LCMS-based Biochemical Assay Protocol

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LCMS grade formic acid, methanol, acetonitrile, water, dimethyl sulfoxide (DMSO), sodium phosphate buffer, and absolute ethanol were supplied by Thermo Fisher Scientific (Waltham, MA, USA). Other chemicals including p-nitrophenyl-α-D-glucopyranose (PNPG), α-glucosidase enzyme, α-amylase enzyme, glycine, 3,5-dinitrosalicylic acid (DNS), potato starch, and sodium phosphate buffered saline (PBS) were purchased from Sigma Aldrich (St. Louis, MO, USA). All the standard compounds including acarbose, quercetin, daidzein, quercitrin, kaempferol, formononetin, 6-hydroxyflavanone, and geniposide were purchased from Acros Organics (Geel, Belgium) with >98% purity.
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