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8 protocols using sodium acetate

1

Comprehensive Analysis of Bioactive Compounds

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The following reagents and solvents were used: acetonitrile and 98–100% formic acid acquired from Merck (Darmstadt, Germany); the standards of phenolic compounds and iridoids were purchased from Extrasynthese (Genay, France); the standards of sugars and organic acids, ferrous chloride (FeCl3), 2,4,6-tri(2-pyridyl)-s-triazine (TPTZ), 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), potassium persulfate, and 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox) were purchased from Sigma Chemical Co. (Steinheim, Germany); methanol and hydrochloric acid were obtained from POCh (Gliwice, Poland); and hydrochloric acid, acetic acid, sodium carbonate, and sodium acetate were purchased from Chempur (Piekary Śląskie, Poland). All reagents were of analytical or HPLC grade.
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2

Analytical Method for Sulfonamides

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Sulfadiazine (SDZ), sulfaguanidine (SGD), sulfamethazine (SMZ), sulfamerazine (SMR), sulfamethoxazole (SMO), and fluorescamine were obtained from Sigma Aldrich (St. Louis, MO, USA). HPLC-grade acetonitrile, methanol, and acetone were purchased from Baker (Deventer, The Netherlands). Ethyl acetate, acetic acid (99.5%), and sodium acetate were purchased from Chempur (PiekaryŚląskie, Poland), and the ammonia solution 25% was purchased from POCH (Gliwice, Poland). Purified water was prepared in-house with a Milli-Q water system from Millipore (Bedford, MA, USA).
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3

Analytical Determination of Tartrazine

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Tartrazine (certified analytical
standard) was purchased from the Institute of Dyes and Organic Products
(Zgierz, Poland). Metafen and Poltram Combo Forte were purchased from
Polpharma (Starogard Gdański, Poland). Sodium acetate (analytical
purity grade) was purchased from Chempur (Piekary Śla̧skie,
Poland). Methanol (for HPLC, super gradient) was purchased from POCH
(Gliwice, Poland). Water used in all experiments was purified using
an HLP demineralizer from Hydrolab (Gdańsk, Poland). Glass-backed
HPTLC RP-18 W plates were purchased from Merck (Darmstadt, Germany).
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4

Polyphenol Quantification and Antioxidant Assays

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Cyanidin 3 O-glucoside (Cy3-glc) was purchased from Extrasynthese (Lyon Nord, France). 5 O-caffeoylquinic acid (5-CQA, chlorogenic acid), quercetin 3 O-rutinoside, (+)-catechin, sugar standards, 1,1-diphenyl-2-picrylhydrazyl radical (DPPH); 2,4,6-tri(2-pyridyl)-s-triazine (TPTZ), 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox); acetonitrile (gradient grade for HPLC), methanol, and formic acid (98–100%) were acquired from Sigma-Aldrich (Germany, Schnelldorf). Organic acids were purchased from Supelco Analytical (Bellefonte, PA, USA). Sulfuric acid, Folin–Ciocalteu reagent, hydrochloric acid, acetic acid, sodium carbonate and sodium acetate were purchased from Chempur (Piekary Śląskie, Poland).
All reagents were of analytical grade.
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5

Chitosan-Lignin Composite Synthesis

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Medium molecular weight chitosan (Sigma-Aldrich), alkali lignin, low sulfonate content (Sigma-Aldrich), citric acid monohydrate (≥99.0%, STANLAB), titanium(iv) isopropoxide (97+%, Sigma-Aldrich), 2-propanol (99.7%, Alfa Aesar), nitric acid (65%, Alfa Aesar), acetonitrile (99.9%, POCH), methanol (99.9%, POCH), benzyl alcohol (99.8%, Sigma-Aldrich), 1,4-benzoquinone (98%, Sigma-Aldrich), dimethylsulfoxide (>99.5%, ROTH), sodium acetate (99.0% Chempur), sodium sulfate (>99%, ROTH), orthophosphoric acid (85%, POCH), potassium trioxalatoferrate(iii) trihydrate (98%, abcr), Nafion (∼5% in a mixture of lower aliphatic alcohols and water, Sigma-Aldrich), 1,10-phenanthroline (99.5%, Chempur) and active carbon Norit® SX 2 (POCH) were used without further purification. Water used was purified to 18 MΩ cm resistivity by Milli-Q water purification system.
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6

Trace Analysis of Organotin Compounds

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Pure standards of tributyltin chloride TBT (96%), triphenyltin chloride TPhT (95%) and internal standard deuterated tributyltin chloride–d27 TBT-d27 (96%) were purchased from Sigma Aldrich (St. Louis, MO, USA). HPLC gradient–grade methanol, acetonitrile and formic acid 98% were purchased from Merck (Darmstadt, Germany). Quechers salts: Magnesium sulphate, sodium chloride, sodium acetate, ammonium acetate were purchased from Chempur (Piekary Śląskie, Poland), trisodium citrate was purchased from Avantor (Gliwice, Poland) and disodium citrate was purchased from Acros Organics (Morris Plains, NJ, US). Sorbents, silica functionalized with primary–secondary amine (PSA) and octadecyl groups were purchased from Agilent Technologies (Santa Clara, CA, USA). Certificated Reference Material BCR® 646 was purchased from Merck (Darmstadt Germany). Ultrapure water was obtained from a Millipore water purification system (MiliQ, Billerica, MA, US) equipped with UV-lamp (resistivity of 18.2 MΩ.cm (at 25 °C) and a TOC value below 5 ppb).
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7

Antioxidant Activity Evaluation Protocol

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The following reagents were bought from Sigma-Aldrich (St. Louis, MO, USA): 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS); 2,2-diphenyl-1-picrylhydrazyl (DPPH); 2,4,6-Tris(2-pyridyl)-s-triazine (TPTZ); acetonitrile; aluminium trichloride (AlCl3); apigenin; ascorbic acid; bovine serum albumin (BSA); caffeic acid; chloramine T; gallic acid; linoleic acid; PBS; potassium persulfate; quercetin; and rutin, trolox and thiobarbituric acid. The following were purchased from CHEMPUR (Piekary Śląskie, Poland): 96% ethanol (46.07 g/mol, 0.808 g/mL); Arnov’s reagent; hydrochloric acid (HCl); iron sulfate (FeSO4); methanol; and sodium acetate (CH3COONa). Reagents such as calcium carbonate (CaCO3), iron trichloride (FeCl3) and sodium carbonate (Na2CO3) were manufactured by POCH (Gliwice, Poland). Folin–Ciocalteau reagent and trichloroacetic acid were manufactured by Eurochem BGD (Tarnów, Poland). Sodium hydroxide (NaOH) was purchased from STANLAB (Lublin, Poland) and p-coumaric acid from Fluka (Chemie AG, Buchs, Switzerland). Orthophosphoric acid (H3PO4; 85%) was delivered by Penta (Prague, Czech Republic).
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8

Antioxidant Activity Assessment Protocol

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2,2-diphenyl-1-picrylhydrazyl (DPPH), Folin-Ciocalteu reagent, 3,4,5-trihydroxybenzoic acid (Gallic acid) and 6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid (Trolox) were purchased from Sigma-Aldrich (Munich, Germany). Ethanol, potassium chloride and sodium carbonate were purchased from Avantor Performance Materials (Gliwice, Poland). Sodium acetate was purchased from Chempur (Piekary Śląskie, Poland). All chemicals and reagents were of analytical grade.
The freeze-drying process was carried out using a TG15 lyophilizer by an external company Naturim (Włocławek, Poland) . The entire extraction process used a Vibra-Cell ultrasonic processor (130 Watt, 20 kHz). Spectrophotometric assays were performed using a V-770 double-beam Jasco spectrophotometer.
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