The stachyose (pure>80%) was extracted from soybean residue [22 (link)]. Soybean isoflavones (pure>95%) includes 50% of daidzin, 6% of daidzein, 26% of glycitin, 3% of glycitein, 10% of genistin and 2% of genistein for animal experiments and the analytical standards of daidzein, glycitein and genistein (pure>98%) were bought from Shanghai Yuanye Biotechnology co., Ltd. (Shanghai, China). Assay kits of total cholesterol (TC), total triglyceride (TG), high-densitylipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), glutamic-oxal(o)acetic transaminase (GOT) and glutamicpyruvic transaminase (GPT) were purchased from HuiliBiotechnology Co. Ltd. (Changchun, China). Additionally, enzyme linked immunosorbent assay ELISA kits of interleukin-1 (IL-1), tumor necrosis factor (TNF-α), nuclear factor kappa-B (NF-κB), acetyl CoA carboxylase (ACC), fatty acid synthetase (FAS), glucose, insulin, leptin and lipopolysaccharide (LPS) were purchased from Jiancheng Bioengineering Institute (Nanjing, China). All other reagents and chemicals were analytical grade or higher.
Daidzein
Manufactured by Yuanye Bio-Technology
Sourced in China
Daidzein is a phytoestrogen compound found in various plants, including soybeans. It is a type of isoflavone with a core chemical structure of 4',7-dihydroxyisoflavone. Daidzein is commonly used as a reference standard and analytical tool in laboratory settings.
Automatically generated - may contain errors
Lab products found in correlation
Daidzin, by Yuanye Bio-Technology (11 mentions)
Genistin, by Yuanye Bio-Technology (8 mentions)
Genistein, by Yuanye Bio-Technology (8 mentions)
Puerarin, by Yuanye Bio-Technology (5 mentions)
Glycitin, by Yuanye Bio-Technology (4 mentions)
Glycitein, by Yuanye Bio-Technology (4 mentions)
Quercetin, by Yuanye Bio-Technology (3 mentions)
Formic acid, by Tianjin Kermel (3 mentions)
Ethanol, by Xilong (3 mentions)
Methanol, by Xilong (3 mentions)
Formic acid, by Xilong (3 mentions)
Ethylene glycol, by Xilong (3 mentions)
Epicatechin, by Yuanye Bio-Technology (2 mentions)
Ferulic acid, by Yuanye Bio-Technology (2 mentions)
Acetonitrile, by Thermo Fisher Scientific (2 mentions)
Rutin, by Yuanye Bio-Technology (2 mentions)
3 hydroxypuerarin, by Yuanye Bio-Technology (2 mentions)
Ononin, by Yuanye Bio-Technology (2 mentions)
3 methoxypuerarin, by Yuanye Bio-Technology (2 mentions)
Naringenin, by Yuanye Bio-Technology (2 mentions)
16 protocols using daidzein
1
Extraction and Purification of Soybean Compounds
2
Soybean Fermentation with R. oligosporus
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The high purity (≥97%) standards, including ascorbic acid, vanillic acid, syringic acid, epicatechin, daidzin, glycitin, ferulic acid, genistin, daidzein, glycitein, quercetin, and genistein were purchased from Yuanye Bio-Technology Co., Ltd. (Shanghai, China). Other reagents of analytical grade. Soybeans (Liaodou, yellow seed coat, mainly planted in northern China, weigh 21.6 g of one hundred seeds) were obtained from a local market. R. oligosporus RT-3 isolated, screened and identified from Tempeh in our laboratory was used for soybean fermentation (11 (link), 12 ). The strain was cultivated on potato dextrose agar at 30°C for 3 days. Then, spores produced were scraped in water (1 × 107 spores/mL).
3
Phytochemical Analysis of Pueraria Extracts
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Folin-Ciocalteu's reagent, 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox), 2,2′-Azobis(2-methylpropionamidine) dihydrochloride (AAPH), fluorescein sodium salt, 2′,7′-dichlorofluorescin diacetate (DCFH-DA), Tris(hydroxymethyl)methyl aminomethane, β-Nicotinamide Adenine Dinucleotide (β-NAD), β-Mercatoethanol (β-Me) and α-D-glucopyranoside (pNPG) were obtained from Sigma-Aldrich (St. Louis, MO, USA). The enzymes used, including alcohol dehydrogenase (ADH) from saccharomyces cerevisiae, potassium-activated aldehyde dehydrogenase (ALDH) from yeast, α-glucosidase from saccharomyces cerevisiae, and α-amylase from porcine pancreas were all purchased from Sigma-Aldrich (St. Louis, MO, USA). Standard phenolic compounds including 3′-hydroxypuerarin, 6″-O-xylosidepuerarin, puerarin, mirificin, 3″-methyoxypuerarin, daidzin, rutin, ferulic acid, genistin, phlorizin, ononin, daidzein, genistein, quercitrin, and p-coumaric acid were obtained from Yuanye Biotechnology Co., Ltd. (Shanghai, China). HPLC-grade acetic acid and acetonitrile were purchased from Fisher Scientific (Waltham, MA, USA). All other reagents used were of analytical grade.
4
Quantitative Analysis of Pueraria lobatae Radix Constituents
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The ILs tested
in the present study are given inTable S1 in the Supporting Information and used without further purification.
High-performance liquid chromatography (HPLC) grade acetonitrile were
obtained from Tedia Company Inc., USA. All other chemicals were of
analytical grade (Tianjin Kermel Chemical Reagent Co., Ltd, China),
and ultrapure water (Milli-Q system) was used throughout.
Analytical
standards of 3′-hydroxypuerarin, puerarin, 3-methoxypuerarin,
daidzin, and daidzein (purity ≥98%) were supplied by Shanghai
Yuanye Biotechnology Co., Ltd, China. P. lobatae Radix in dried bulk form was obtained from the local drugstore in
Jinan, China. It was pulverized in a grinder (Zhongxing Weiye, Beijing,
China), passed through 20–40 mesh sieves, and the sample was
stored at room temperature.
in the present study are given in
High-performance liquid chromatography (HPLC) grade acetonitrile were
obtained from Tedia Company Inc., USA. All other chemicals were of
analytical grade (Tianjin Kermel Chemical Reagent Co., Ltd, China),
and ultrapure water (Milli-Q system) was used throughout.
Analytical
standards of 3′-hydroxypuerarin, puerarin, 3-methoxypuerarin,
daidzin, and daidzein (purity ≥98%) were supplied by Shanghai
Yuanye Biotechnology Co., Ltd, China. P. lobatae Radix in dried bulk form was obtained from the local drugstore in
Jinan, China. It was pulverized in a grinder (Zhongxing Weiye, Beijing,
China), passed through 20–40 mesh sieves, and the sample was
stored at room temperature.
5
Isoflavone Mixture Effects on PC12 Cells
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An SI mixture containing 51.81% daidzin, 19% glycitin, 7.94% genistin, 2.45% daidzein, 0.81% glycitein, and 0.52% genistein, and their standards, were obtained from Shanghai Yuanye Bio-Technology Co., Ltd. (Shanghai, China) [15 (link)]. Penicillin/streptomycin, fetal bovine serum (FBS), and Dulbecco’s modified Eagle medium (DMEM) were purchased from Gibco (CA, USA). PC12 cell lines were purchased from Nanjing Keygen Biotech. Co., Ltd. (Nanjing, China). The kits for ROS, apoptosis, the cell cycle, and the protein content were obtained from the Shanghai Beyotime Institute of Biotechnology (Shanghai, China). Antibodies for β-tubulin, Bax, cyclin A, p21, Bcl-xL, hypoxia-inducible factor-1 alpha (HIF-1α), LC3, p62, Nrf2, p-Akt, Akt, p-mTOR, mTOR, p-p38 MAPK, p38 MAPK, and LamB were obtained from Shanghai Abmart Inc. (Shanghai, China).
6
Bacterial Strains and Reagents for Biotransformation
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E. coli DH5α and C41(DE3) were used as hosts for DNA cloning and whole-cell biotransformation, respectively. Primer STAR HS DNA polymerase and restriction endonucleases were obtained from Takara (Dalian, China). DNA and genomic DNA Extraction Kits were obtained from Thermo Scientific (Waltham, United States) and TIANGEN (Beijing, China), respectively. The plasmid miniprep purification kit was purchased from Sangon Biotech (Shanghai, China). Oligonucleotide synthesis and sequence analysis were achieved by Sangon Biotech (Shanghai, China). ALA and hemin were purchased from Sigma-Aldrich (St. Louis, MO, United States). Naringenin, eriodictyol, dihydrokaempferol, dihydroquercetin, kaempferol, quercetin, apigenin, luteolin, daidzein, and 7,3′,4′-trihydroxyisoflavone were purchased from Yuanye Bio-Technology (Shanghai, China). Other chemicals were purchased from Sangon Biotech (Shanghai, China) and were of the highest commercial grade available.
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E. coli DH5α and C41(DE3) were used as hosts for DNA cloning and whole-cell biotransformation, respectively. Primer STAR HS DNA polymerase and restriction endonucleases were obtained from Takara (Dalian, China). DNA and genomic DNA Extraction Kits were obtained from Thermo Scientific (Waltham, United States) and TIANGEN (Beijing, China), respectively. The plasmid miniprep purification kit was purchased from Sangon Biotech (Shanghai, China). Oligonucleotide synthesis and sequence analysis were achieved by Sangon Biotech (Shanghai, China). ALA and hemin were purchased from Sigma-Aldrich (St. Louis, MO, United States). Naringenin, eriodictyol, dihydrokaempferol, dihydroquercetin, kaempferol, quercetin, apigenin, luteolin, daidzein, and 7,3′,4′-trihydroxyisoflavone were purchased from Yuanye Bio-Technology (Shanghai, China). Other chemicals were purchased from Sangon Biotech (Shanghai, China) and were of the highest commercial grade available.
7
UPLC-MS/MS Analysis of Isoflavonoids
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An Ultimate 3000 Ultra High Performance Liquid Chromatography (UPLC) system coupled with LTQ-Orbitrap velos pro mass spectrometer was used (Thermo-Fisher, Waltham, MA, USA), as well as an Acquity UPLC I-Class UPLC system coupled with Xevo TQ-S micro triple quadrupole mass spectrometer (Waters, Milford, MA, USA).
The reference compounds of formononetin, ononin, daidzein, daidzin, genistin, genistein, calycosin-7-O-β-d -glucoside, biochanin A, and chlorogenic acid (purity > 98% for all) were all purchased from Shanghai Yuanye Biotechnology Co., Ltd. (Shanghai, China). Acetonitrile and formic acid were purchased from Fisher Scientific, San Jose, CA, USA Company (mass spec-grade). Deionized water was prepared by Milli-Q ultrapure water preparation system (Millipore Co., Ltd., Billerica, MA, USA). All other chemicals were purchased from Beijing Chemical Works (Beijing, China, analytical grade).
The reference compounds of formononetin, ononin, daidzein, daidzin, genistin, genistein, calycosin-7-O-β-
8
Soybean Phosphatidyl Choline Lipid Formulation
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Soybean phosphatidyl choline (SPC) was purchased from Lipoid GmbH (Germany). Cholesterol and DSPE-mPEG2000 were purchased from AVT Pharmaceutical Co., Ltd. (Shanghai, China). HPLC-grade methanol and acetonitrile were purchased from TEDIA Company (USA). Chloroform and methanol (analytical grade) were obtained from Sinopharm Chemistry Reagent Co., Ltd. (Shanghai, China). The water was purified by Milli-Q® water purification system ((Millipore, USA). Daidzein (≥ 98% in purity) was purchased from Yuanye Biotechnology Co., Ltd. (Shanghai, China). Apigenin (internal standard, IS, ≥ 98% in purity) was purchased from Delge Pharmaceutical Technology Co., Ltd. (Nanjing, China). Phosphotungstic acid hydrate (analytical grade) was purchased from Macklin Co., Ltd. (Shanghai, China). Tween-80 and ethyl acetate were obtained from Sigma (Missouri, USA). Formic acid (MS-grade) was purchased from Fischer (USA).
9
Quantification of Pueraria lobata Compounds
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Reference standards of puerarin, 3′-methoxypuerarin, daidzin and daidzein (purity ≥ 98%) were supplied by Shanghai Yuanye Biotechnology Co., Ltd, China. Methanol and formic acid (Analytical Grade, Tianjin, China) were purchased from Tianjin Kermel Chemical Reagent Co., Ltd. Acetonitrile (Tedia Company Inc., Fairfield, USA) was used for HPLC analysis. Deionized water (>18 MΩ cm) was prepared using an Elga Purelab water system (Elga, England).
Dry Puerariae lobata was purchased from the Zhonglu Hospital. The roots of Puerariae lobata were washed, dried and ground in a grinder (Zhongxing Weiye, Beijing, China). The powder was passed through a 60 mesh sieve and stored at −4 °C.
Dry Puerariae lobata was purchased from the Zhonglu Hospital. The roots of Puerariae lobata were washed, dried and ground in a grinder (Zhongxing Weiye, Beijing, China). The powder was passed through a 60 mesh sieve and stored at −4 °C.
10
Fatty Acid and Sterol Profiling
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The sesame oil, rapeseed oil, and soybean oil were purchased from local markets (Wuhan, Hubei, China). Before being used for analysis, all samples were stored in the dark. The standard samples of fatty acid methyl ester (FAME), brassicasterol, ergosterol, campesterol, campestanol, stigmasterol, β-sitosterol, Δ5-avenasterol, cycloartanol, cycloartenol, 24-methylene-cycloartenol, and cholestanol were purchased from the Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). Daidzin, daidzein, genistin, and genistein were purchased from the Shanghai Yuanye Biotechnology Co. (Shanghai, China). Methyl sterculate (C20H36O2, Purity ≥ 98 %) and methyl malvalate (C19H34O2, purity ≥ 95 %) were purchased from the Matreya Lipid and Biochemistry Technology Co., Ltd., USA. All solvents used were of analytical grade, and all standards were stored at −20 °C in darkness.
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