2,2-Diphenyl-1-picrylhydrazyl (DPPH•), Potassium persulfate, the Folin–Ciocalteu reagent, gallic acid and high purity standards were purchased from Sigma-Aldrich (Madrid, Spain). Methanol, acetonitrile, formic acid, anhydrous sodium carbonate and sodium acetate were supplied from Scharlau Chemie S.A. (Sentmenat, Spain). All reagents and solvents were purchased from Sigma-Aldrich (Madrid, Spain) with the exception of Methanol (Honeywell, Germany). Authentic standards of gallic acid, salvianolic acid, caffeic acid, hesperidine, luteolin, rosmarinic acid, apigenin, genkwanin, quercetin and olivetol were obtained from Sigma-Aldrich (Madrid, Spain).
Genkwanin
Manufactured by Merck Group
Sourced in United States
Genkwanin is a laboratory equipment product offered by Merck Group. It functions as a tool for research and analysis purposes. The core function of Genkwanin is to facilitate specific laboratory procedures and experiments, but a detailed description cannot be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Quercetin, by Merck Group (4 mentions)
Apigenin, by Merck Group (4 mentions)
Luteolin, by Merck Group (3 mentions)
Kaempferol, by Merck Group (3 mentions)
Rosmarinic acid, by Merck Group (2 mentions)
Methanol, by Merck Group (2 mentions)
Ferulic acid, by Merck Group (2 mentions)
Isoquercitrin, by Merck Group (2 mentions)
Folin ciocalteu reagent, by Merck Group (1 mentions)
Caffeic acid, by Merck Group (1 mentions)
Salvianolic acid, by Merck Group (1 mentions)
Olivetol, by Merck Group (1 mentions)
Gallic acid, by Merck Group (1 mentions)
Allopurinol, by Merck Group (1 mentions)
Isoferulic acid, by Merck Group (1 mentions)
Xanthine substrate, by Merck Group (1 mentions)
Febuxostat, by Merck Group (1 mentions)
Techtochrysin, by Merck Group (1 mentions)
Ro 61 8048, by Merck Group (1 mentions)
Lc ms grade water, by Avantor (1 mentions)
6 protocols using genkwanin
1
DPPH Antioxidant Capacity Assay
2
Xanthine Oxidase Inhibition Assay
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XO enzyme (grade I, from bovine milk, approximately 10.4 units per mL) and xanthine substrate were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Allopurinol, febuxostat, quercetin, rosmarinic acid, luteolin, kaempferol, isoferulic acid, genkwanin and techtochrysin (analytical grade) were also obtained from Sigma-Aldrich Co. All other reagents and solvents used in the study were of analytical grade.
3
Enzymatic Assay for Kynurenine Metabolites
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3′-Hydroxy-alpha-naphthoflavone and 3′-Hydroxy-ss-naphthoflavone were bought from Indofine (Township, NJ, USA), Genkwanin, Apigenin,Ro 61–8048, and DSMOwere obtained from Sigma-Aldrich Co. LLC (Germany). KMO kit and buffers were bought from (Thermo Fisher Scientific (Waltham, MA, USA)).
4
HPLC-MS/MS Quantification of Alfalfa Metabolites
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Chromatographic separation for quantitative analysis was carried out by the same HPLC-MS/MS system mentioned above. Briefly, 1 μL solution was injected into the column. The flow rate was set to 0.2 mL min−1 and the column oven temperature to 25 °C ± 1 °C. The mobile phase consisted of water (A) and methanol (B) with the following gradient profile: 0–2 min, 95% A; 2–20 min, 100% B; 20–22 min, 100% B; 22–23 min, 95% A and 23–30 min, 95% A.
Based on the literature describing typical alfalfa secondary metabolite profile and screening of mixed Hunor-40 BJ 16 molecules were selected. For quantitative analysis nicotinamide (≥99.5%); nicotinic acid (≥98%); riboflavin (≥98%); isoquercitrin (primary reference standard); naringenin (≥95%); biochanin A; coumestrol (≥95%); formononetin (≥98%); luteolin (≥98%); apigenin (≥98%); liquiritigenin (phyproof Reference Substance); tricin (phyproof Reference Substance); Apigenin 7-O-glucuronide (primary reference standard); biotin (≥99%); quercetin (≥95%); genkwanin (≥98%); ferulic acid (≥99%) as external standards were applied (Merck-Sigma, Darmstadt, Germany). Working standard solutions were prepared daily by dilution of the stock solutions with methanol.
Based on the literature describing typical alfalfa secondary metabolite profile and screening of mixed Hunor-40 BJ 16 molecules were selected. For quantitative analysis nicotinamide (≥99.5%); nicotinic acid (≥98%); riboflavin (≥98%); isoquercitrin (primary reference standard); naringenin (≥95%); biochanin A; coumestrol (≥95%); formononetin (≥98%); luteolin (≥98%); apigenin (≥98%); liquiritigenin (phyproof Reference Substance); tricin (phyproof Reference Substance); Apigenin 7-O-glucuronide (primary reference standard); biotin (≥99%); quercetin (≥95%); genkwanin (≥98%); ferulic acid (≥99%) as external standards were applied (Merck-Sigma, Darmstadt, Germany). Working standard solutions were prepared daily by dilution of the stock solutions with methanol.
5
Quantitative analysis of phytochemicals
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For the chemical analysis, HCl (analytical reagent grade) (a.r.), 37%, NaOH (≥97.0%) LC–MS grade water, and a 3kDa PES membrane filter were ordered from VWR International, Radnor, USA. The AccQ-Tag Ultra derivatization reagent kit, a mixture of standard proteinogenic amino acids, the AccQ-tag Ultra eluent A and AccQ-tag Ultra eluent B were purchased from Waters (Waters, Milford, MA, USA). The gradient grade methanol (MetOH) (≥99.9%) and standards of phytochemical compos: nicotinamide (≥98%), nicotinic acid (a. s.), biotin (≥99%), riboflavin (a. s.), liquiritigenin (≥97% (HPLC), formononetin (a. s.), chlorogenic acid (analytical standard), neochlorogenic acid (≥98%), chryptochlorogenic acid (a. s.), syringaldehyde (98%), sulforaphane (≥90% synthetic, liquid), sinapic acid (≥98%), scopoletin (analytical standard), apigenin (≥95.0%); apigenin-7-O-glucuronide (primary reference standard); luteolin (≥98%); quercetin (≥95.0%); isoquercitrin (a. s.); naringenin (≥95.0%); genkwanin (≥98%); kaempferol (≥97.0% (HPLC); isoliquiritigenin (a. s.), and ferulic acid (USP reference standard) were obtained from Sigma-Aldrich (Darmstadt, Germany).
6
Cultivating and Treating Human Respiratory Cells
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Normal human bronchial epithelial cells (NHBE, Lonza, Basel, Switzerland) were purchased from Lonza (Cat. No. CC-2450) and were maintained in Keratinocyte SFM basal medium supplement with 5 μg/L human recombinant epithelial growth factor (EGF), 50 mg/L Bovine Pituitary Extract (BPE), 5 mg/L insulin and 25 nM hydrocortisol. Cells were passaged every 3 days and plated for experiment treatment within 6 passages. Human alveolar type II epithelia cells A549 cell was purchased from ATCC (Cat.No.CCL-185) and were maintained in DMEM medium supplement with 10% FBS, sodium pyruvate, and 1% penicillin/streptomycin. Cells were maintained at 37 °C in a 5% CO2 humidified atmosphere. CuSO4, chloroquine, N-acetylcysteine, fusin, apigenin, kaempferol, and genkwanin were purchased from Sigma-Aldrich (St. Louis, MO, USA).
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