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3 protocols using h2so4

1

Evaluation of Antioxidant and Antimicrobial Activities

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OFL, ENR, and SAR (Table 1) of high purity (>99%)
and catalase (lyophilized powder from bovine liver 2000–5000
units/mg protein) were purchased from Sigma-Aldrich. Na2SO3 anhydride (85–90%), FeSO4·7H2O, H2O2 (33% w/v), ascorbic acid, 1,10-phenanthroline
1-hydrate, H2SO4 (96% w/w), and UHPLC-grade
methanol and acetonitrile were obtained from AppliChem-Panreac. Formic
acid (80% w/w) was purchased from VWR Chemicals. Ultrapure water was
prepared with a Merck Milli-Q system. The zone of inhibition tests
was made using E. coli CECT 101 strain,
employing tryptone soya and Mueller–Hinton broth, both provided
by Scharlau.
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2

Quantification of Fluoroquinolone Antibiotics

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High purity (>99%) FLU, OA, OFL, ENR, and CIP were purchased from Sigma-Aldrich. NaOH, H2SO4 (96%), glacial acetic acid, and UHPLC-grade methanol and acetonitrile were provided by AppliChem-Panreac. Ammonium acetate was provided by Scharlau.
Respective (F)Q 250 µM stock solutions were prepared in basic media; all solutions were stable at dark conditions (no hydrolysis was observed).
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3

Analytical Method for Antimicrobial Compounds

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Thiabendazole (TBZ), flumequine (FLU), oxolinic acid (OA), ofloxacin (OFL), enrofloxacin (ENR) and ciprofloxacin (CIP) (molecular structures shown in Fig. 1) high purity (>99 %), gum Arabic, lignin sulphonic acid sodium, peptone, and humic acid substances were purchased from Sigma-Aldrich. Beef extract from Bovril. FeSO 4 •7H 2 O, H 2 O 2 (33 % w/v), NaHCO 3 , NaCl, (NH 4 ) 2 SO 4 , NaOH, acetic acid (96 %), 1,10-phenanthroline 1-hydrate, H 2 SO 4 (96 %), and UHPLC grade methanol and acetonitrile, were obtained from AppliChem-Panreac. Ammonium acetate, K 2 HPO 4 , MgSO 4 •7H 2 O, CaSO 4 •2H 2 O and ascorbic acid were purchased from Scharlau, and formic acid (80 %) from VWR Chemicals. Ultra-pure water (MQ) was prepared with a Merck Milli-Q system. FLU, OA, OFL, ENR and CIP 300 μM stock solutions were prepared in basic media and TBZ 300 μM in acidic media. All the solutions were stable when preserved in the dark, and no hydrolysis was observed.
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