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2 protocols using trimethoprim d9

1

Pharmaceutical Target Compounds Analysis

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Based on the marketing data of pharmaceuticals, 57 types of pharmaceuticals were initially selected as target compounds in our preliminary study. Out of the 57 compounds, we selected 27 compounds as the targets in this study based on their influent concentrations and detection frequencies. In total 27 types of pharmaceuticals, including 11 antibiotics, 6 anti-inflammatory/NSAIDs, 2 antiarrhythmic drugs, and 1 stimulant were selected as the target substances. Table 1 shows the physicochemical properties of the substances. For the standard substances and reagents used in the analysis, 10 mg/L of a standard solution of 98% or higher purity purchased from Sigma-Aldrich (Saint Louis, MO, USA) and Fluka (Seelze, Germany) was prepared and stored in a freezer at −20 °C or below. It was diluted and used on the day of the analysis. Twenty types of isotopically labeled standards (acetaminophen-d4, salicylic acid-d4, atenolol-d7, caffeine-13C3, carbamazepine-d10, cimetidine-d3, ciprofloxacin-d8, diclofenac-13C6, diphenhydramine-d3, erythromycin-d6, gemfibrozil-d6, ibuprofen-d3, iopromide-d3, ketoprofen-d3, naproxen-d3, ofloxacin-d3, propranolol-d7, sulfadimethoxine-13C6, testosterone-d3, and trimethoprim-d9) were purchased from Sigma-Aldrich and Toronto Research Chemicals (Toronto, ON, Canada).
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2

Quantification of Sulfamethoxazole and Trimethoprim

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Sulfamethoxazole, trimethoprim and trimethoprim-D9 were obtained from Sigma-Aldrich (MO, USA). Sulfamethoxazole-N-acetyl and its internal standard Sulfamethoxazole-N-acetyl-D4 were purchased from Santa Cruz (TX, USA). Sulfamethoxazole-D4 was obtained from Alsachim (Illkirch Graffenstaden, France). The chemical structures of Sulfamethoxazole, Sulfamethoxazole-D4, Sulfamethoxazole-N-acetyl, Sulfamethoxazole-N-acetyl-D4, trimethoprim and trimethoprim-D9 are shown in Figure 1. Ammonium acetate and acetic acid were retrieved from Merck (NJ, USA). Trifluoroacetic acid and acetonitrile LC-MS were both obtained from Biosolve (Dieuze, France). Water was in house purified using a Milli-Q system (Millipore Corporation, MA, USA).
Sulfamethoxazole-D4 was used as internal standard to quantify Sulfamethoxazole. Sulfamethoxazole-Nacetyl was quantified using Sulfamethoxazole-N-acetyl-D4 and trimethoprim-D9 was used as internal standard for the trimethoprim quantification. The internal standard solution contained 200 ng/ml Sulfamethoxazole-D4, 2000 ng/ml Sulfamethoxazole-N-acetyl-D4 and 10 ng/ml trimethoprim-D9 dissolved in methanol (Merck, NJ, USA).
Buffer solution used in the gradient elution consisted of ammonium acetate (5.0 g/l), acetic acid (100%, 35 ml/l) and Trifluoroacetic acid (100%, 2 ml/l).
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