Quercetin dihydrate

Manufactured by Merck Group

Sourced in United States, Germany, Italy, India

Quercetin dihydrate is a crystalline compound that is a form of the flavonoid quercetin. It is a white to light-yellow powder that is used as a laboratory reagent.

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Quercetin dihydrate powder Quercetin

55 protocols using quercetin dihydrate

Quercetin Modulation of Cell Signaling

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Dulbecco’s modified Eagle’s media (DMEM), Dulbecco’s phosphate buffer saline (DPBS), fetal bovine serum (FBS), penicillin–streptomycin, quercetin dihydrate (Qu), 3-(4,5-dimetylthiazol-yl)-diphenyl tetrazolium bromide (MTT), propidium iodide, rhodamine 123, 2,7-dichlorodihydrofluoresceindiacetate (H₂DCF-DA), Fluo-3 AM, proteinase K, ribonuclease A, 1,2-bis-(O-aminophenoxy)-ethane-N,N,N’,N’-tetraacetic acid, tetraacetoxymethyl ester (BAPTA-AM), β-nicotinamide adenine dinucleotide reduced disodium salt (β-NADH), ascorbic acid, agarose, dimethyl sulfoxide (DMSO) and anti-β-actin antibody were purchased from Sigma–Aldrich Chemicals (St. Louis, MO). Antibodies against Bcl-2, Bax, Bim, AIF, NF-κB, caspase-3/caspase-8, phospho-MEK, MEK1/2, phospho-ERK, ERK 1/2, phospho-p38, p-38, phospho-JNK, JNK, Nrf-2, catalase, Cu/Zn SOD, PI3K, phospho-Akt, Akt and GAPDH were purchased from Santa Cruz Biotechnology (Santa Cruz Biotechnology, Inc,). FITC Annexin V was purchased from BD Pharmingen. Histone H3 antibody was purchased from Cell Signaling Technology.
Stock solution of quercetin (Qu) was prepared in dimethyl sulfoxide (DMSO) and diluted to the desired final concentration with culture medium just before use. The final DMSO concentration did not exceed 0.1% (v/v).

Rafiq R.A., Quadri A., Nazir L.A., Peerzada K., Ganai B.A, & Tasduq S.A. (2015). A Potent Inhibitor of Phosphoinositide 3-Kinase (PI3K) and Mitogen Activated Protein (MAP) Kinase Signalling, Quercetin (3, 3', 4', 5, 7-Pentahydroxyflavone) Promotes Cell Death in Ultraviolet (UV)-B-Irradiated B16F10 Melanoma Cells. PLoS ONE, 10(7), e0131253.

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Cellular Uptake of Pharmaceutical Agents

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Quercetin dihydrate, phenolsulfonphthalein (PSP), probenecid, rifampicin, vincristine, Hank's balanced salts, minimum essential medium (MEM) non-essential amino acid solution, poly-L-lysine hydrobromide, D-glucose, HEPES, and sodium bicarbonate were purchased from Sigma (St. Louis, MO). Docetaxel trihydrate was obtained from Shin Poong Pharmaceutical Co. Ltd. (Ansan, Republic of Korea). Zoletil® 50 (a mixture of 25 mg/ml of zolazepam and tiletamine each) was obtained from Virbac (Carros, France). All other chemicals and reagents were of analytical or HPLC grade as appropriate.

Oh J.H., Lee J.H, & Lee Y.J. (2019). Evaluation of the Mrp2-mediated flavonoid-drug interaction potential of quercetin in rats and in vitro models. Asian Journal of Pharmaceutical Sciences, 14(6), 621-630.

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Plant Extract Analytical Protocols

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Ethyl alcohol and acetone, used for the extraction process, were obtained from Sigma Aldrich (Steinheim, Germany). The materials used for the analysis of the extracts were Folin Ciocalteu phenol reagent (2N) obtained from Merck (Darmstadt, Germany), sodium sufhate anhydrous (>99%) from Mallinckrodt (St. Louis, MS, USA), 2,2-diphenyl-1-picryl hydrazyl (DPPH) from Sigma-Aldrich (Steinheim, Germany) and gallic acid (98% w/w) from Acrōs Organics (Fair Lawn, NJ, USA). The standard compounds used in the study were quercetin dihydrate and rosmarinic acid, products of Sigma-Aldrich (Steimheim, Germany). Carnosic acid was obtained from Dayang Chemicals Co (Hangzhou, China), and carnosol from Extrasynthese (Lyon, France). Water, acetonitrile, methanol, and trifluoroacetic acid for LC-MS analyses were obtained from Fisher Chemical (Leicestershire, UK).

Psarrou I., Oreopoulou A., Tsimogiannis D, & Oreopoulou V. (2020). Extraction Kinetics of Phenolic Antioxidants from the Hydro Distillation Residues of Rosemary and Effect of Pretreatment and Extraction Parameters. Molecules, 25(19), 4520.

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HPLC Standards for Biochemical Analysis

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All solvents used were of HPLC grade and supplied by Fisher Scientific. Canavanine sulfate salt, quercetin-3-glucoside, quercetin dihydrate, hyperoside, luteoloside, stachydrine (hydrochloride), trigonelline chloride, diosmetin, H-Glu-Tyr-OH and H-Phe-Glu-OH standards were purchased from the Sigma-Aldrich (Munich, Germany), Thermo scientific, Carl Roth (Karlsruhe, Germany), Chemodex (St. Gallen, Switzerland), Cayman (Düsseldorf, Germany) and Adipogen life sciences (San Diego, CA, USA) companies.

Abouzeid S., Beutling U., Elekhnawy E, & Selmar D. (2023). Antibacterial and Antibiofilm Effects of Allelopathic Compounds Identified in Medicago sativa L. Seedling Exudate against Escherichia coli. Molecules, 28(6), 2645.

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Prostate Cancer Cell Line Culture

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Human PCa cell lines LNCaP, DU-145, and PC-3 were obtained from American Type Culture Collection (ATCC). LNCaP, DU-145, and PC-3 cells were cultured in Roswell Park Memorial Institute (RPMI), 1640 medium at 37 °C with 5% CO₂ and supplemented with 10% fetal bovine serum (FBS; Hyclone, Logan, UT, USA). Normal prostate epithelial cells (PrEC), with materials purchased from ATCC, were cultured in basal medium with cell growth kit containing the following: 6 mM L-glutamine, 0.4% Extract P, 1.0 μM epinephrine, 0.5 ng/mL rhTGFα, 100 ng/mL hydrocortisone, 5 μg/mL rh insulin, and 5 μg/mL Apo-transferrin. All the cell lines were checked and confirmed as mycoplasma-free. Quercetin dihydrate (Sigma Aldrich, St. Louis, MO, USA) was dissolved in 100% dimethyl sulfoxide (DMSO; Corning, Manassas, VA, USA) before further dilutions. Working concentrations did not exceed DMSO of 0.2%.

Ward A.B., Mir H., Kapur N., Gales D.N., Carriere P.P, & Singh S. (2018). Quercetin inhibits prostate cancer by attenuating cell survival and inhibiting anti-apoptotic pathways. World Journal of Surgical Oncology, 16, 108.

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Antioxidant Activity Characterization

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All standards and reagents were of analytical grade. Sodium acetate, sodium carbonate, sodium hydroxide, potassium chloride, potassium persulfate, catechin hydrate, gallic acid, fluorescein sodium salt, quercetin dihydrate, 2,2′-azobis (2-amidinopropane) dihydrochloride (AAPH), 2,2-azinobis(3-ethylbenzthiazoline-6-sulphonic acid) diammonium salt (ABTS), 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2′,7′-dichlorofluorescein diacetate (DCFH-DA), Folin-Ciocalteu reagent, 6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid (Trolox), dimethyl sulfoxide (DMSO), iso-quercitrin, 4-OH benzoic acid, rutin, trans-ferulic acid, trans-p-coumaric acid, vanillic acid, vitexin, acetonitrile (HPLC grade), ethanol, hydrochloric acid, methanol (HPLC grade), phosphoric acid (ACS grade), trichloroacetic acid, cell culture media and supplements were purchased from Sigma-Aldrich (St. Louis, MO, USA). Aluminum chloride and sodium nitrite were purchased from Carlo Erba (Milan, Italy). A Simplicity 185 purification system (Millipore SAS, Molsheim, France) was used to treat double deionized water (ddH₂O; 18.2 MΩ/cm, 20 °C).

Gabriele M., Cavallero A., Tomassi E., Arouna N., Árvay J., Longo V, & Pucci L. (2024). Assessment of Sourdough Fermentation Impact on the Antioxidant and Anti-Inflammatory Potential of Pearl Millet from Burkina Faso. Foods, 13(5), 704.

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Isoflavones and Liver Microsome Interactions

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Isoflavone kaempferol (3,5,7-trihydroxy-2-(4-hydroxyphenyl)-4H-1-benzopyran-4-one, 98% purity), and genistein (4’,5,7-trihydroxy isoflavone, 98%), and quercetin dihydrate (3,3’,4’,5,7-Pentahydroxyflavone dihydrate, 98%) were from Sigma-Aldrich (St. Louis, MO). Human liver microsomes (HLMs) from pooled individual human donors’ liver, ferrous chloride tetrahydrate (FeCl₂∙4H₂O, purity, 99%), phosphate-buffered saline (PBS, 0.1 M), dimethyl sulfoxide (DMSO, 99% purity) and H₂O₂ were purchased from Fisher Scientific Suwanee, GA. Double deionized water was purified using a Milli-Q system (Millipore Corporation, MA).

Boadi W.Y., Stevenson C., Johnson D, & Mohamed M.A. (2021). Flavonoids Reduce Lipid Peroxides and Increase Glutathione Levels in Pooled Human Liver Microsomes (HLMs). Advances in biological chemistry, 11(6), 283-295.

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Comprehensive Chemical Reagents Protocol

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Sodium bicarbonate, potassium persulphate, sodium acetate, acetic acid and hydrochloric acid were purchased from Qualigens (Mumbai, India), and 2,2-azinobis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), and methanol from Merk (Darmstadt, Germany). Sodium chloride was procured from HiMedia Laboratories (Mumbai, India). 2,2-Diphenyl-1-picryhydrazyl (DPPH), ascorbic acid, and all polyphenolic standards (i.e. rutin hydrate, phloridzin dihydrate, p-coumaric acid, (+)-catechin hydrate, gallic acid, quercetin dihydrate, 3-hydroxybenzoicacid, 4-hydroxybezoicacid, ellagic acid, vanillic acid, caffeic acid, m-coumaric acid, ferulic acid, trans-cinnamic acid and chlorogenic acid) and alkaloids (i.e. berberine and palmatine) were procured from Sigma Aldrich (St. Louis, Missouri, United States). All chemicals were of analytical or HPLC grade and the solutions were prepared with methanol and lab ultrapure water (Rions India Lab Water Systems, India).

Belwal T., Pandey A., Bhatt I.D, & Rawal R.S. (2020). Optimized microwave assisted extraction (MAE) of alkaloids and polyphenols from Berberis roots using multiple-component analysis. Scientific Reports, 10, 917.

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Antioxidant Capacity of L. rhinocerotis Extracts

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The antioxidant capacity of L. rhinocerotis extracts was evaluated based on methods previously reported (below); hence, only the necessary modifications will be indicated. Standards including quercetin dihydrate, 6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid (Trolox), ferrous sulphate heptahydrate (FeSO₄·7H₂O), and disodium ethylenediamine tetraacetate (Na₂EDTA) were obtained from Sigma-Aldrich (St. Louis, USA), while 1,1,3,3-tetraetoxypropane (TEP) (the tetraethylacetal of malondialdehyde [MDA]) was purchased from Merck (Darmstadt, Germany). Other chemicals and solvents used were of analytical grade. All extracts were dissolved in 50% (v/v) methanol in water to produce stock solutions of 20 mg/ml and diluted to desired concentrations for the following assays:

Lau B.F., Abdullah N., Aminudin N., Lee H.B., Yap K.C, & Sabaratnam V. (2014). The Potential of Mycelium and Culture Broth of Lignosus rhinocerotis as Substitutes for the Naturally Occurring Sclerotium with Regard to Antioxidant Capacity, Cytotoxic Effect, and Low-Molecular-Weight Chemical Constituents. PLoS ONE, 9(7), e102509.

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Evaluation of Rabbit Muscle Glycogen Phosphorylase

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Glycogen phosphorylase α from rabbit muscle, glycogen from rabbit liver (type III), α-D glucose-1-phosphate, HEPES [4-(2-hydroxyethyl) piperazine-1-ethanesulfonic acid, N-(2-hydroxyethyl) piperazine-N′-(2-ethanesulfonic acid)], magnesium chloride (MgCl2), EGTA (ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid), ammonium molybdate, malachite green, caffeine and potassium chloride, butylated hydroxyanisole (BHA), 2,2 diphenyl-1-picrylhydrazyl (DPPH), 2,4,6-tripyridyl-s-triazine (TPTZ), gallic acid monohydrate, ferrozine, sodium nitroferricyanide (III) dehydrate, sodium nitrite, Griess reagent, curcumin, polyvinyl polypyrrolidone (PVP), sodium acetate trihydrate, sodium phosphate dibase, sodium phosphate monobasic, and quercetin dihydrate were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Ascorbic acid, acetic acid glacial, sodium chloride, ferrous sulfate (FeSO₄), ferric chloride hexahydrate (FeCl₃·6H₂O), ethylenediaminetetraacetic acid disodium dehydrate (EDTA-Na₂·2H₂O), Folin-Ciocalteu phenol reagent, and sodium carbonate were purchased from Merck Chemical Co. (Malaysia). HPLC grade water was purchased from Fisher Scientific (Malaysia). All chemicals used are of analytical grade and were used without further purifications.

Ablat A., Mohamad J., Awang K., Shilpi J.A, & Arya A. (2014). Evaluation of Antidiabetic and Antioxidant Properties of Brucea javanica Seed. The Scientific World Journal, 2014, 786130.

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