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8 protocols using ch3oh

1

Biodiesel Production from Used Cooking Oil

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UCO utilized for the experiment
was obtained from an eatery in Dehradun, India. The procedure used
in pretreating the oil sample and its physicochemical properties have
been previously reported.7 (link) All the chemical
compounds (KOH, CH3OH, NaHSO3, Al2O3, and NaOH) herein were provided by Thermo-Fisher Scientific
Industries, Mumbai, India.
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2

Preparation of Ionic Soluble IBP

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The required amount of IBP powder (200 mg) was dissolved in 10% methanol (CH3OH; Thermo Fisher Scientific) and dissolved in DI water in a 250 mL volumetric flask. It was then stirred for 12 h, sonicated for 2 h, and filtered using a 0.45 μm pore-size cellulose acetate membrane filter. A sodium chloride (NaCl) solution was added to the filtrate to create ionic strength (0.01 M) in the solution. The pH was adjusted to 7 using a sodium phosphate (Na3PO4) solution.
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3

Zirconium Tetrachloride and L-Aspartic Acid Synthesis

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Zirconium tetrachloride (ZrCl4, 99.5%) and L-aspartic acid (C4H7NO4, LA, 99.0%) were obtained from Strem Chemicals Inc. (Newburyport, MA, USA) and Sigma Aldrich Co., Ltd. (St. Louis, MO, USA), respectively. Deionized water was used as the solvent for the preparation of all chemicals solutions. Ethanol (99.9%, CH3OH) was purchased from Fisher Scientific. All chemicals obtained were used directly without further purification.
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4

Electrospun PCL Scaffold Production

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PCL (Mn 80 000 g/mol, Sigma Aldrich Chemistry, 440 744) scaffolds were produced using 12 wt% polymer in 1:7.3 chloroform (CHCl3, Sigma Aldrich, ReagentPlus, 132950) to methanol (CH3OH, Fisher Chemicals, HPLC grade, CAS: 67‐56‐1) volume ratio solvent. IME Technologies EC‐CLI electrospinning equipment was used with a translation nozzle stage and an 8 cm diameter rotating drum collector with a 19 cm spinning distance rotating at 10 rpm. The environmental parameters were controlled at 24°C and 40% relative humidity. A needle with inner diameter of 0.8 mm and outer diameter of 1 mm was used. The source voltage was set to 24 kV and collector voltage to −2 kV. The flowrate of the polymer solution was 26 μL/min. The mats that were produced were cut into smaller pieces of 20 by 10 mm for application. The scaffolds were sterilized using a cold ethylene oxide (EO) gas procedure and finally kept under vacuum for 5 days before use to remove EO residue.
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5

Synthesis and Characterization of Chitosan Derivatives

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The natural polysaccharide, chitosan (C6H11NO4)n, was gained from Sigma-Aldrich (USA). Other reagents, such as phenyl acrylic acid (PA, PhCH = CHCOOH, 99.0%), dimethyl amine (DMA, (CH3)2NH, 40.0%), thionyl chloride (SOCl2, 99.0%), pyridine (C5H5N, 99.5%), and triethyl amine (C6H15N:98.5%), were procured from Darmstadt, Germany, and Loba Chemie, India, respectively. Solvents, including dichloromethane (CH2Cl2, 99.0%), methanol (CH3OH, 99.7%), and acetone (CH3COCH3, > 99.5%), were purchased from Fisher Chemicals (United States) and El-Nasr companies (Egypt), respectively. Distilled water was prepared in our research lab. All reagents and solvents for synthesis were of analytical grade and used without extra purification.
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6

Analytical Reagent Protocols

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All the chemicals used in this work were of analytical reagent grade and procured from Sigma-Aldrich (Burlington, MA, USA) and Fluka (Buchs, Switzerland) chemical companies. Folin–Ciocalteu reagent, quercetin, and gallic acid standards were provided by Sigma-Aldrich Co. (St. Louis, MO, USA). AlCl3 hexahydrate, sodium carbonate, and CH3OH were collected from Fisher Scientific (Fair Lawn, NJ, USA). A Milli-Q system (Millipore Corporation, Bedford, NH, USA) was used to produced purified water.
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7

HPLC Mobile Phase Preparation

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Acetonitrile, CH3CN (HPLC grade, ≥99.9%, SIGMA-ALDRICH®, St. Louis, MO, USA), methanol, CH3OH (HPLC grade, ≥99.9%, Fisher Chemical, Waltham, MA, USA), orthophosphoric acid, H3PO4 (HPLC, 85–90%, FLUKA®, Seelze, Germany) were used in the preparation of the mobile phase. Nylon membrane filters of 0.45 μm pore size were used in solvent filtration (HNWP, MERCK MILLIPORE® Ltd., Billerica, MA, USA). Ultra-pure (HPLC grade) water was obtained using a Milli-Q water purification system.
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8

Detailed Analytical Procedures for Chemical Synthesis

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All chemicals were obtained from commercial sources and used without further
purification unless otherwise noted. Anhydrous DMF, CH3OH, DMSO and EtOH
were purchased from Fisher Scientific. Anhydrous THF, acetone, CH2Cl2, CH3CN, and
ether were obtained using a solvent purification system (mBraun Labmaster 130). NMR
solvents were purchased from Cambridge Isotope Laboratories (Andover, MA). All 1H,
13C, 19F and 31P NMR spectra were obtained either on a JEOL ECX 400 MHz NMR,
operated at 400 and 100 MHz, respectively, or a Bruker AVANCE III HD 500 MHz NMR,
operated at 500 and 125 MHz, respectively, and referenced to internal
tetramethylsilane (TMS) at 0.0 ppm. The spin multiplicities are indicated by the
symbols s (singlet), d (doublet), dd (doublet of doublets), t (triplet), q
(quartet), m (multiplet), and br (broad). Reactions were monitored by thin-layer
chromatography (TLC) using 0.25 mm Whatman Diamond silica gel 60-F254 pre-coated
plates. Purification was performed on a Teledyne Isco CombiFlash Rf 200,and eluted
with the indicated solvent system. Yields refer to chromatographically and
spectroscopically (1H and 13C NMR) homogeneous materials. Mass
Spectra were recorded at the UMBC MCAC for nominal using Bruker APOLLO™ II
ESI/APCI - MALDI Dual Source for apex(R)-Qe FTMS or Johns Hopkins Mass Spectrometry
Facility for high resolution using VG Analytical VG-70SE Magnetic Sector Mass
Spectrometer.
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