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4 protocols using quercetin dihydrate

1

Metabolite Extraction and Purification

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Ammonium formate, 98% crystalline (Alfa Aesar, United States), EDTA ≥ 98.5% w/w (Sigma-Aldrich, United States), methanol HPLC-grade (Thermo Fisher Scientific, United States), ultrapure grade water purified to 18.1 MΩ⋅cm using a Barnstead water filtration system (Thermo Fisher Scientific, United States), potato starch (Sigma; 102954), and quercetin dihydrate, 97% w/w (Alfa Aesar, United States).
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2

Synthesis of Kyoto Green Fluorescent Dye

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Kyoto Green was synthesized following the procedure described in Ojida et al.23 (link). Magnesium chloride (MgCl2), zinc nitrate (Zn(NO3)2), HEPES minimum 99.5% titration (C8H18N2O4S), p-nitrophynyl thymidine 5′-monophosphate (pNph-5′-TMP), sodium pyrophosphate decahydrate (NaPPi), adenosine 5′-triphosphate disodium salt (ATP), and adenosine 5′-monophosphate disodium salt (AMP) were products of Sigma-Aldrich (St. Louis, MO, USA) or Merck (Darmstadt, Germany). Cordycepin, uridine, cytidine, inosine, thymidine, kaempferol, 2′-deoxyadenosine, 2′-deoxyinosine, 2′-deoxyuridine, and 2′-deoxyguanosine were purchased from Fujifilm Wako Pure Chemical Corporation (Japan). Myricetin, quercetin dihydrate and guanosine were commercially available from Alfa Aesar (UK). Recombinant human ENPP1 was purchased from R&D systems (Minneapolis, USA).
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3

Quercetin and Iodine Nanoformulation Development

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Quercetin dihydrate (QCT) and iodine were purchased from Alfa Aesar (Haverhill, MA, USA); Pluronic® P 123 (P123), Kolliphor® TPGS (TPGS), Pluronic® L 92 (L92), and Kolliphor P 407 (P407) were gifted by BASF Corporation (Florham Park, NJ, USA). DMEM media, RPMI 1640 media, trypsin, DMSO, and PBS were procured from Mediatech Inc. (Manassas, VA, USA). Tween 80 and MTT were obtained from Sigma-Aldrich Co. (St Louis, MO, USA). Potassium iodide was obtained from Ameresco (Solon, OH, USA). In all the experiments, Millipore ultrapure water was used.
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4

Quercetin Dihydrate Solubilization Protocol

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Quercetin dihydrate with a purity of 97%
was obtained from Alfa Aesar (Port of Heysham Industrial Park, Lancashire,
England). Dimethyl sulfoxide (DMSO) solvent was purchased from Fisher
Scientific (Bishop Meadow Road, Loughborough, England), and ethanol
solvent (99.98%) was purchased from VWR Chemicals. Water purified
by treatment with a Milli-Q apparatus was used.
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