The heavy metals used in these experiments were cadmium (II) chloride, copper (II) chloride, and lead (II) chloride, which are major contaminants of soil and water surrounding urbanized and industrialized locations, in agricultural regions, and near mining and hydraulic fracturing sites2 (link),3 (link),5 (link),27 (link),40 (link). The metals were found to accumulate in floral tissues of plants grown in soil contaminated with these metals7 (link). For cadmium (II) chloride (99.99%; Fisher Scientific, Hampton, NH) and lead (II) chloride (99%; Acros Organics, Morris, NJ), the metal concentrations used were 0.001 mg/L, 0.01 mg/L, 0.1 mg/L, 1 mg/L, and 10 mg/L. For copper (II) chloride (99%; Fisher Scientific, Hampton, NH), the metal concentrations used were 0.002 mg/L, 0.02 mg/L, 0.2 mg/L, 2 mg/L, and 20 mg/L. Each metal was diluted to the desired concentrations in a 1 M sucrose solution. These concentrations are comparable to or less than the concentrations of these metals found in contaminated environments and measured in the floral parts of plants grown in contaminated soils7 (link),13 (link). Additionally, Di, et al.29 (link) showed these concentrations to be sublethal to adult honey bees in survival assays29 (link).
Copper 2 chloride
Manufactured by Thermo Fisher Scientific
Sourced in Belgium, United States, Croatia
Copper (II) chloride is a chemical compound with the formula CuCl2. It is a crystalline solid that is soluble in water and other polar solvents. Copper (II) chloride has various applications in analytical chemistry, electroplating, and the production of other copper compounds.
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Lab products found in correlation
Quercetin, by Merck Group (8 mentions)
Sodium carbonate, by Kemika (7 mentions)
Trolox, by Merck Group (7 mentions)
2 4 6 tri 2 pyridyl s triazine tptz, by Thermo Fisher Scientific (6 mentions)
Neocuproine, by Thermo Fisher Scientific (6 mentions)
Orthophosphoric acid hplc grade 85, by Thermo Fisher Scientific (4 mentions)
Potassium persulfate, by Kemika (4 mentions)
Rutin, by Merck Group (4 mentions)
Cyanidin 3 glucoside, by Extrasynthese (4 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (3 mentions)
Gallic acid, by Merck Group (3 mentions)
Caffeic, by Merck Group (3 mentions)
Chlorogenic, by Merck Group (3 mentions)
Phlorizin, by Merck Group (3 mentions)
Phloretin, by Merck Group (3 mentions)
P coumaric, by Merck Group (3 mentions)
Folin ciocalteu reagent, by Kemika (3 mentions)
Ellagic acid, by Merck Group (3 mentions)
Methanol, by Avantor (3 mentions)
Silver nitrate, by Merck Group (3 mentions)
22 protocols using copper 2 chloride
1
Heavy Metal Accumulation in Plant Tissues
2
Preparation of Buffered Metal Solutions
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Sulfuric acid (ACS/FCC, BDH Aristar, 95.0–98.0%) was acquired from VWR (Pittsburgh, PA, USA). Sodium bicarbonate and hexa-ammine ruthenium(iii ) chloride from Sigma-Aldrich (St. Louis, MO, USA). Copper(ii ) sulfate, copper(ii ) chloride, copper(ii ) acetate, cobalt(ii ) acetate, and silver nitrate were purchased from Fisher Scientific (Fair Lawn, NJ, USA). All aqueous solutions were prepared using 18 MΩ cm water (NANOpure Diamond, Barnstead; Dubuque, IA) and analytical reagent grade chemicals. Phosphate buffer solution was prepared by dissolving anhydrous Na2HPO4 (Fisher Scientific; Fair Lawn, NJ, USA) in ultrapure water. The pH of the solutions was measured using a glass electrode connected to a digital pH meter (Orion 420A+, Thermo; Waltham, MA, USA) and adjusted with 1 mol L−1 solutions of either NaOH or HCl.
3
Silica-Based Affinity Chromatography
4
Detailed Preparation of Enriched Compounds
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The 3D-enriched fragment library was purchased from Life Chemicals and divided into mixtures of 4–5 fragments as previously described [11 (link)]. Amino acids, uracil, thiamine-HCl, LB broth, biotin, and nicotinic acid were purchased from RPI Corp. The 5-fluoroindole, magnesium sulfate, succinic acid, calcium chloride, Iron(III) chloride, 15N ammonium chloride, L-Moses, and imidazole were purchased from Millipore Sigma. Potassium diphosphate, potassium monophosphate, sodium phosphate, manganese (II) chloride, zinc(I) chloride, and sodium chloride were purchased from Fischer Scientific. Cobalt (II) chloride, Ethylenediaminetetraacetic acid, and copper (II) chloride was purchased from Acros Organics. Boric acid was purchased from Mallinckrodt. Deuterium oxide and dimethyl sulfoxide-d6 were purchased from Cambridge Isotope Laboratories. GSK4027 was purchased from Cayman Chemicals and TP238 was synthesized previously [55 (link)].
5
Antioxidant Potential of Protein Matrices
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Almond protein matrix was purchased from Raab Vitalfood GmbH (Rohrbach, Germany) and brown rice protein matrix was from Kernnel premium (Zagreb, Croatia). Quercetin, trolox and 2,2-diphenyl-1-picrylhydrazyl (DPPH) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Sodium carbonate was obtained from Kemika (Zagreb, Croatia). Neocuproine, copper (II) chloride and 2,4,6-tri(2-pyridyl)-s-triazine (TPTZ) were obtained from Acros Organics (Geel, Belgium). Orthophosphoric acid (HPLC grade > 85%) was obtained from Fisher Scientific (Loughborough, UK), while methanol (HPLC grade) was purchased from J.T. Baker (Deventer, The Netherlands). Iron (III) chloride hexahydrate, ethanol, sodium acetate and ammonium acetate were purchased from Gram-mol (Zagreb, Croatia).
6
Analytical Characterization of Therapeutic Protein Solution
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DTPA, EDTA, glutathione (GSH), quercetin, coumarine-3-carboxylic acid (CCA), 7-(diethylamino)coumarin-3-carbohydrazide (CHH), desferrioxamine, ascorbate, potassium phosphate (KH2PO4), and multi-compendial grade polysorbate 20 (PS20) and PS80 were from Sigma-Aldrich (Merck KgaA, Darmstadt, Germany). Copper(II) chloride and iron(III) chloride were from Acros Organics (Thermo Fischer Scientifics, Waltham, MA, USA). Ultra-gradient HPLC grade acetonitrile was from J.T. Baker (Avantor Performance Materials, Radnor, PA, USA). Liquid chromatography-MS grade methanol was from Merck KgaA (Darmstadt, Germany). Ultrapure water was from a Millipore Milli-Q water system (Merck KgaA, Darmstadt, Germany). The therapeutic protein solution was provided by Lek d.d. (Mengeš, Slovenia).
7
Antioxidant Capacity of Apple and Citrus Fibers
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Apple fibers were obtained from Biesterfeld AG (Zagreb, Croatia) and citrus fibers were obtained from Fiberstar (River Falls, WI, USA). Gallic acid, 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), trolox, 2,2-diphenyl-1-picrylhydrazyl (DPPH), and quercetin were obtained from Sigma-Aldrich (St. Louis, MO, USA). Neocuproine, 2,4,6-tri(2-pyridyl)-s-triazine (TPTZ), and copper (II) chloride were purchased from Acros Organics (Geel, Belgium). Methanol (HPLC grade) was obtained from J.T. Baker (Deventer, Netherlands) and orthophosphoric acid (HPLC grade > 85%) was obtained from Fisher Scientific (Loughborough, UK). Iron (III) chloride hexahydrate, sodium acetate, ethanol, and ammonium acetate were purchased from Grammol (Zagreb, Croatia).
8
Antioxidant and Anti-Diabetic Potential of Pectin and Apple Fiber
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The apple fiber powder was supplied by Biesterfeld AG (Zagreb, Croatia) and both types of pectin were the product of CP Kelco (Atlanta, GA, USA). The 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), trolox, 4-(dimethylamino)-cinnamaldehyde (DMAC), chlorogenic, caffeic, p-coumaric, gallic and ellagic acids, rutin, quercetin, phloretin and phlorizin were purchased from Sigma-Aldrich (St. Louis, MO, USA). The α-glucosidase (from Saccharomyces cerevisiae) and acarbose were obtained from Sigma-Aldrich (St. Louis, MO, USA). The 4-nitrophenyl-α-D-glucopyranoside was obtained from Alfa Aesar (Kandel, Germany) and potassium dihydrogen phosphate was from BDH Prolabo (Poole, UK). Potassium persulfate, Folin-Chiocalteu reagent and sodium carbonate were obtained from Kemika (Zagreb, Croatia). Neocuproine, 2,4,6-tri(2-pyridyl)-s-triazine (TPTZ) and copper (II) chloride were purchased from Acros Organics (Geel, Belgium). Methanol (HPLC grade) was from J.T. Baker (Deventer, The Netherlands) and orthophosphoric acid (HPLC grade > 85%) was from Fisher Scientific (Loughborough, UK). Iron (III) chloride hexahydrate, sodium acetate, ethanol, ammonium acetate and starch were bought from Gram-mol (Zagreb, Croatia). Cyanidin 3-glucoside was from Extrasynthese (Genay, France).
9
Comprehensive Phytochemical Analysis of Apple Fibers
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Apple fibers were obtained from Biesterfeld AG (Zagreb, Croatia). 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), 4-(dimethylamino)-cinnamaldehyde (DMAC), trolox, 2,2-diphenyl-1-picrylhydrazyl (DPPH), rutin, quercetin, phloretin, phlorizin, chlorogenic, caffeic, p-coumaric, gallic, ellagic acids, myrtenol and α-amylase (from the porcine pancreas) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Potassium persulfate, Folin–Ciocalteu reagent and sodium carbonate were purchased from Kemika (Zagreb, Croatia). Neocuproine, 2,4,6-tri(2-pyridyl)-s-triazine (TPTZ) and copper (II) chloride were bought from Acros Organics (Geel, Belgium). Methanol (HPLC grade) was from J.T. Baker (Deventer, Netherlands) and orthophosphoric acid (HPLC grade > 85%) was from Fisher Scientific (Loughborough, UK). Iron (III) chloride hexahydrate, sodium acetate, ethanol, ammonium acetate and starch were bought from Gram-mol (Zagreb, Croatia). Cyanidin 3-glucoside was from Extrasynthese (Genay, France) and 3,5-dinitrosalicylic acid from Alfa Aesar (Kandel, Germany).
10
Citrus Fiber Antioxidant Potential Analysis
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Citrus fiber (76.1% of dietary fiber, while the rest are proteins—7%, sugars—7.2%, fats—0.77%, minerals—3.06%, and moisture—5.85%) was obtained from Fiberstar (River Falls, WI, USA). Potassium persulfate and sodium carbonate were purchased from Kemika (Zagreb, Croatia). Neocuproine, hesperidin, 2,4,6-tri(2-pyridyl)-s-triazine (TPTZ) and copper (II) chloride were bought from Acros Organics (Geel, Belgium). Trolox, 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), 4-(dimethylamino)-cinnamaldehyde (DMAC), quercetin, caffeic acid, p-coumaric acid, chlorogenic acid, rutin, gallic acid and ellagic acid were obtained from Sigma-Aldrich (St. Louis, MO, USA). Cyanidin 3-glucoside was from Extrasynthese (Genay, France). Methanol (HPLC grade) was from J.T. Baker (Deventer, Netherlands), and orthophosphoric acid (HPLC grade > 85%) was from Fisher Scientific (Loughborough, UK). Iron (III) chloride hexahydrate, sodium acetate, ethanol, ammonium acetate and Folin–Ciocalteu reagent were bought from Gram-mol (Zagreb, Croatia).
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