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3 protocols using tilmicosin

1

Synthesis and Characterization of Antimicrobial Polymer Matrices

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AZM, ERY, and tilmicosin (TM) were obtained from Sigma-Aldrich (USA). Oleandomycin (OLE), tylson (TYL), and clarithromycin (CLA) were obtained from Dr. Ehrenstorfer (Germany). Dopamine (DA), streptomycin (STM), gentamicin (GM), tetracycline (TC), neomycin (NM), and thiamphenicol (TAP) were purchased from Probe Biotechnology LLC (China). The chemical molecular structures of these substances are shown in Fig 1. AZM-dispersible tablets (250 mg) were obtained from a local pharmacy. Ethylene glycol dimethacrylate (EGDMA), methacrylic acid (MAA) and 2,2-azobisisobutyronitrile (AIBN) were obtained from Sigma-Aldrich (USA). In order to remove the polymerization inhibitor, MAA and EGDMA were purified by distillation in vacuum under nitrogen protection then stored at 4°C before use. AIBN was recrystallized from hot methanol and dried in a vacuum. AB (purity > 99.99%) was obtained from Strem Chemicals (USA). Paraffin oil and graphite powder were obtained from Sinopharm Chemical Reagent Co., Ltd. (China). Acetonitrile and methanol were purchased from KeMiOu Chemical Reagent Co. (China). All other chemicals were of analytical grade and obtained from Sinopharm Chemical Reagent Co., Ltd. (China). All solutions were freshly prepared with ultrapure water.
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2

Analytical Standards for Tylosin and Tilmicosin

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All chemicals and solvents were used with analytical purity. Analytical standards of tylosin and tilmicosin (>95% purity) were obtained from Sigma-Aldrich (St. Louis, MO, USA), also methanol and acetonitrile (HPLC >95% purity), orthophosphoric acid (H 3 PO 4 85%) from Merck (Darmstadt, Germany), and Disodium ethylenediaminetetraacetate (Na 2 EDTA) from Sigma Aldrich (St. Louis, MO, ABD). Ultrapure water was obtained using the H2OPRO-VF-T/Arium Ultrapure (Sartorius, Germany) device.
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3

Antibiotic Stock Solution Preparation

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The following antibiotics were purchased from Sigma-Aldrich (St. Louis, MO): β-lactams: ampicillin (AMP), penicillin G (PENG), amoxicillin (AMO), cloxacillin (CLO), dicloxacillin (DIC), penicillin V (PENV), oxacillin (OXA), nafcillin (NAP), ceftiofur (CEFT), cefuroxime (CEFM), cefalexin (CEFA), cefoperazone (CEFZ), and cefazolin (CEFL); tetracyclines: doxycycline (DOX), tetracycline (TET), oxytetracycline (OXY), and chlortetracycline (CTE); aminoglycosides: streptomycin (STR), neomycin (NEO), dihydrostreptomycin (DSTR), gentamicin (GEN), kanamycin (KAN), and spectinomycin (SPE); sulfonamides: sulfamonomethoxine (SMM), sulfadimidine (SDM), sulfadiazine (SDZ), sulfamethoxypyridazine (SMP), sulfaquinoxaline (SQX), and sulfamethoxazole (SMX); macrolides: erythromycin (ERY), tylosin (TYL), tilmicosin (TIL), spiramycin (SPI); and lincosamides: lincomycin (LIN), trimethoprim (TMP), and chloramphenicol (CAP). The components used in the preparation of stock solutions and working solutions of antibiotics are shown in Table 1. Stock solutions of antimicrobial agents (1 mg/mL) were prepared in suitable solvents and stored at -20°C in amber glass vials for up to 2 wk. Working standard solutions (1 µg/mL) were prepared by suitable dilutions of the stock solutions until use.
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