title: Evaluation of in vitro biotransformation of propranolol with HPLC, MS/MS, and two bioassays.
abstract: The majority of human drugs enter aquatic systems after ingestion and subsequent excretion in the form of parent compounds and metabolites. Environmental exposure to drug metabolites has not been reported so far. The goal of the present study was to apply the in vitro method of biotransformation of compounds with S9 fraction to the ecotoxicological analysis. beta-adrenoceptor antagonist propranolol was metabolized with S9 rat liver fraction. The parent compound was quantified with HPLC, and the metabolites were identified with QToF MS. Propranolol was metabolized rapidly, during the first hour its level decreased by 80 and 50% of the initial 20 and 100 mg L(-1), respectively. Ten peaks were observed on the HPLC-RF chromatogram. Four peaks were identified with QToF MS/MS propranolol (m/z = 260), N-desisopropylpropranolol (m/z = 218), hydroxypropranolol (m/z = 276), and hydroxy N-desisopropranolol glycol (m/z = 235). Then the ecotoxicity of the reaction mixture was studied with two bioassays Spirotox with the protozoan Spirostomum ambiguum and Thamnotoxkit F with the anostracean crustacean Thamnocephalus platyurus. Propranolol is twofolds more toxic to Spirotox than to Thamnotoxkit F with 24 h-EC50 = 1.77 mg L(-1) and 24 h-LC50 = 3.86 mg L(-1), respectively. No statistically significant differences were found between the toxicity of the reaction mixtures after S9 biotransformation and the propranolol solution. These results indicate that the biological activity of the metabolites is similar to that of the parent drug. The presented method of in vitro biotransformation of drugs with S9 fraction followed by HPLC and ecotoxicity tests, may be used as screening method for evaluation of the toxicity of drug metabolites.
authors.affiliation: Department of Environmental Health Sciences, Medical University of Warsaw, Banacha 1 str, 02-097, Warsaw, Poland. grzenaja@wp.pl
authors.forename: Grzegorz
authors.initials: G
authors.lastname: Nałecz-Jawecki
authors.forename: Tomasz
authors.initials: T
authors.lastname: Wójcik
authors.forename: Józef
authors.initials: J
authors.lastname: Sawicki
mesh_terms: D000319:Adrenergic beta-Antagonists / Q000493:pharmacokinetics*; D000818:Animals; D001711:Biotransformation; D002851:Chromatography, High Pressure Liquid; D004784:Environmental Monitoring / Q000379:methods*; D008099:Liver / Q000378:metabolism*; D008297:Male; D013058:Mass Spectrometry; D011433:Propranolol / Q000493:pharmacokinetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D023382:Toxicity Tests, Acute / Q000379:methods
chemical_list: D000319:Adrenergic beta-Antagonists; D011433:Propranolol