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7 protocols using diethyl ether

1

Phytochemical Analysis of E. angustifolia

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Elaeagnus angustifolia leaves and fruits were collected from wild plants from Turkey (Konya) and were botanically and morphologically identified by Dr. Evren Yıldıztugay from the science faculty of Selcuk University, Konya, Turkey. Fruits were in a ready-to-eat ripeness state.
Double-distilled water, ethanol, 98% formic acid, acetonitrile RS, n-hexane for HPLC, and dichloromethane were purchased from Merck Life Sciences s.r.l (Milan, Italy), methanol for HPLC and diethyl ether were purchased from Carlo Erba Reagents (Milan, Italy), DMSO-d6 (99.80% D) was obtained from Eurisotop (Saint-Aubin, France) and CO2 was purchased from Sapio s.r.l. (Monza, Italy).
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2

Antioxidant Activity Evaluation Protocol

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The 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and α-tocopherol were purchased from Alfa Aesar (Karlsruhe, Germany). Sodium chloride, citric acid, lecithin of soya (>97%), diethyl ether, and isooctane were obtained from Carlo Erba (Milano, Italy). Sodium carbonate anhydrous and sodium hydroxide were from Penta (Prague, Czech Republic). Absolute ethanol, methanol, Folin–Ciocalteu reagent, and gallic acid monohydrate were from Panreac (Barcelona, Spain). Cyclohexane and n-hexane were from Sigma-Aldrich (Steinheim, Germany). The deionized water used in the experiments was produced using a deionizing column.
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3

Extraction and Analysis of Bioactive Compounds

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Potassium iodide, glacial acetic acid, chloroform, sodium thiosulfate, sodium hydroxide, starch, β-carotene, α-tocotrienol, α-tocopherol, and α-cholestanol were purchased from Sigma Aldrich (Milan, Italy). Petroleum ether, diethyl ether, ethanol, tetrahydrofuran, n-hexane, citric acid, and sodium sulfate anhydrous were purchased from Carlo Erba (Rodano, Italy). Bleaching earth was purchased from Clariant (Milan, Italy).
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4

Phototherapeutic Nanoparticle Formulation

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SRB free base (molecular weight [MW] 464.8 g·mol−1) was purchased from LC Laboratories, a division of PKC Pharmaceuticals Inc (Woburn, MA, USA). VP (benzoporphyrin-derivative monoacid ring A, MW 718.8 g·mol−1) was kindly supplied by Professor D Dolphin (University of British Columbia, Vancouver, BC, Canada). Poloxamer P123 (EO20–PO65–EO20, MW 5,750 g·mol−1), poloxamer F127 (EO100–PO65–EO100, MW 12,600 g·mol−1), trehalose, polysorbate 80, and Nile red (NR) were purchased from Sigma-Aldrich. Ethanol, dichloromethane, diethyl ether, acetone, and acetonitrile were purchased from Carlo Erba Reagents. All the other chemicals were of analytical reagent grade and used without previous purification.
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5

Solvent Extraction and Analytical Standards

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All solvents, water (LC-MS grade), chloroform, methanol, n-hexane, iso-octane (2,2,4 trimethylpentane), and diethyl ether were acquired from Carlo Erba reagents (Milan, Italy), whereas ethanol absolute (≥99.8%), hydrochloric acid and formic acid (mass spectrometry grade) were purchased from VWR Chemicals (Milan, Italy).
Trichloroacetic acid (TCA, C2HCl3O2), 2-thiobarbituric acid (TBA, C4H4N2O2S), sodium thiosulphate (Na2S2O3), sodium carbonate (Na2CO3), potassium iodide (KI), Folin–Ciocalteu reagent, gallic acid, anhydrous sodium sulfate (Na2SO4) anhydrous sodium sulfate (Na2SO4) and phenolphthalein solution were obtained from Sigma Aldrich (Milan, Italy).
All standards (purity > 98%), namely malondialdehyde (MDA), butylated hydroxytoluene (BHT), gallic acid, α-tocopherol, oleic acid and α-tocopherol were acquired from Sigma Aldrich (Milan, Italy). Chemicals and reagents were of analytical grade.
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6

Polymer Synthesis and Characterization

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D,L-lactide (Lot no.: 2009002968) and glycolide (Lot no.: 1210002433) were purchased from Corbion (Gorinchem, The Netherlands). Tin (II) 2-ethylhexanoate (92.5–100%, Lot no.: 1003287404), poly(ε-caprolactone) diol (PCL, Mn = 2000 g.mol−1, WXBD4206V), 1,3-propanediol (>98%, Lot no.: S768868143) were provided by Sigma Aldrich (St-Quentin Fallavier, France) and used as received. 1,4-Dioxane (ACS-Reagent, Lot no.: M1600), distilled in the presence of CaH2, was provided by Honeywell (Offenbach, Germany). Diethyl ether (Batch Number: V1G060031H), n-pentane (Batch Number: V9C059249D), dichloromethane (Batch Number: V1M602021N) and sodium chloride (Batch Number: V1L077212C) were provided by Carlo Erba Reagents (Val de Reuil, France). Hexamethylene diisocyanate (HDI; >98%, Lot no.: IVRTI-NJ) was provided by TCI Europe (Zwikndrecht, Belgium) and used as received.
Zinc diethyldithiocarbamate (ZDEC, Lot no.: A-223K) and high-density polyethylene film (HDPU, Lot no.: C-221) were provided by Hatano Research Institute, Food and Drug Safety Center (Hadano, Japan).
Dulbecco’s Modified Eagle Medium (DMEM/F-12, Lot no.: RNBL4062), PenicillinStreptomycin (Lot No.: 0000205667) and GlutaMAXTM (Lot no.: 2554717) were purchased from ThermoFisher Scientific (Waltham, MA, USA). Tissue Culture Polystyrene (TCPS) 96-well plates were purchased from Becton Dickinson (Le Pont-de-Claix, France).
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7

Extraction and Concentration of Marigold Oil

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The extracts obtained from the oil extraction need to be concentrated before further analysis. The oil was removed using a saponification method. The saponification method was performed by mixing 5 g of marigold extract with 10 mL of diethyl ether (Labscan, Dublin, Ireland). Then, 10 mL of ethanolic solution containing 10% KOH (Carlo Erba, Cornaredo, Italy) was added into the mixture and incubated at room temperature in the dark for 2 h. The saponifiable part was washed with water and the layer of diethyl ether was dried using a rotary evaporator. The dried sample of the lutein-rich fraction was kept at −20 °C in the dark until further used [21 (link)].
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