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4 protocols using nach3coo

1

Preparing Acetate Buffer Solutions

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Buffer
solutions of NaCH3COO, 0.25 M and pH ca 5, were prepared
from NaCH3COO (Sigma-Aldrich >99%) and CH3COOH
(Sigma-Aldrich >99.7%) dissolved in deionized water (18.2 MΩ
cm).
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2

Cyclic Voltammetry of Melanin Electrodes

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Cyclic voltammetry was performed using a Biologic VSP 300 multichannel potentiostat, with carbon paper current collectors loaded with melanin acting as the working electrode, Pt mesh as the counter electrode and Ag/AgCl(aq) (1 M KCl for Fe/melanin electrodes prepared by pre-immersion and 3 M NaCl for the rest of the work) as the reference electrode. The electrolyte (10 ml) was composed of 145 mM KCH3SO4 (99%, Acros Organics), 10 mM NaCl (≥99%, Sigma Aldrich), 2 mM MgCl2 (≥99%, Sigma Aldrich), 10 mM NaCH3COO (≥99%, Sigma Aldrich), buffered with CH3COOH at pH 7 (Atherton et al., 2008 (link); Kortleven et al., 2011 (link)). The electrolyte at pH 5 was composed of 0.25 M NaCH3COO, buffered with CH3COOH.
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3

Ensis Shells for Capsaicin Encapsulation

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Waste shells of individuals belonging to Ensis species were collected from a local fisher in July 2019 in Izmir, Turkey. Shell samples (24 individuals) were washed repeatedly with distilled water to remove the dust and particles and then dried at room temperature for one week. During the experiment, chemicals used for film isolation; HCl and NaOH were obtained from Merck and Capsaicin (CAS Number: 404-86-4) was purchased from Sigma Aldrich. The chemicals used in Capsaicin loading and release studies (methanol, chloroform and acetic acid) were purchased from Sigma Aldrich, and NaCH 3 COO, Na 2 HPO 4 *12H 2 O, NaH 2 PO 4 *2H 2 O and NaCl from Merck. DPPH (2,2-diphenyl-1picrilhydrazil) (CAS Number: 1898-66-4) used for the antioxidant study was purchased from Sigma-Aldrich. Distilled water was used in all the experiments.
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4

Purification and Characterization of AEMA Monomer

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Monomer 2-aminoethyl methacrylate hydrochloride
(Sigma 90%, AEMA) was dissolved into acetate buffer (100 mg/mL) and
stirred in an Al2O3/inhibitor remover mixture
thoroughly. AEMA was then extracted by centrifuging (15 min, 4025
RCF) the solids out. Chain transfer agents (CTAs) 4-cyano-4-(phenylcarbonothioylthio)pentanoic
acid (Sigma, CPA) and poly(ethylene oxide) methyl ether 2-(dodecylthiocarbonothioylthio)-2-methylpropionate
(Sigma, 6000 g/mol, PEO-CTA) were used as received. All solvents were
used as received except ultrapure deionized water, which was used
fresh from filtration equipment (18 mΩ, UHQ H2O).
Thermal initiator 2,2′-azobis[2-(2-imidazolin-2-yl)propane]dihydrochloride
(Wako Chemicals, VA-044) was recrystallized from methanol with a small
amount of UHQ H2O. 2-Ethyl-2-thiopseudourea hydrobromide
(Sigma), sodium hydroxide (VWR aq. 0.1 M, NaOH), acetic acid (Merck
EMSURE, CH3COOH), hydrochloric acid (Merck EMSURE fuming
or WVR aq. 1 M, HCl), sodium carbonate (Merck EMSURE, NaCO3), sodium bicarbonate (Merck EMSURE, NaHCO3), and sodium
acetate (Merck EMSURE, NaCH3COO) were used as received.
Instrument grade CO2 (Linde, pure) and N2 (Linde,
instrument 5.0, ≥99.999%) were used as received.
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