title: Significance of metabolite extraction method for evaluating sulfamethazine toxicity in adult zebrafish using metabolomics.
abstract: Recently, environmental metabolomics has been introduced as a next generation environmental toxicity method which helps in evaluating toxicity of bioactive compounds to non-target organisms. In general, efficient metabolite extraction from target cells is one of the keys to success to better understand the effects of toxic substances to organisms. In this regard, the aim of this study is (1) to compare two sample extraction methods in terms of abundance and quality of metabolites and (2) investigate how this could lead to difference in data interpretation using pathway analysis. For this purpose, the antibiotic sulfamethazine and zebrafish (Danio rerio) were selected as model toxic substance and target organism, respectively. The zebrafish was exposed to four different sulfamethazine concentrations (0, 10, 30, and 50mg/L) for 72h. Metabolites were extracted using two different methods (Bligh and Dyer and solid-phase extraction). A total of 13,538 and 12,469 features were detected using quadrupole time-of-flight liquid chromatography mass spectrometry (QTOF LC-MS). Of these metabolites, 4278 (Bligh and Dyer) and 332 (solid phase extraction) were found to be significant after false discovery rate adjustment at a significance threshold of 0.01. Metlin and KEGG pathway analysis showed comprehensive information from fish samples extracted using Bligh and Dyer compared to solid phase extraction. This study shows that proper selection of sample extraction method is critically important for interpreting and analyzing the toxicity data of organisms when metabolomics is applied.
authors.affiliation: Bio Monitoring Laboratory, Program in Environmental Technology and Policy, Korea University Sejong Campus, 2511 Sejong-ro, Sejong City, Chungnam 339-700, South Korea.
authors.forename: Ryan
authors.initials: R
authors.lastname: De Sotto
authors.affiliation: Metabolomics Laboratory, College of Pharmacy, Korea University Sejong Campus, 2511 Sejong-ro, Sejong City, Chungnam 339-700, South Korea.
authors.forename: Carl
authors.initials: C
authors.lastname: Medriano
authors.affiliation: Department of Environmental Engineering, Daejeon University, 62 Daehak-Ro, Dong-Gu, Daejeon 300-716, South Korea.
authors.forename: Yunchul
authors.initials: Y
authors.lastname: Cho
authors.affiliation: National Institute of Environmental Research, Environmental Research Complex, Incheon 404-708, Korea.
authors.forename: Kwang-Seol
authors.initials: KS
authors.lastname: Seok
authors.affiliation: Metabolomics Laboratory, College of Pharmacy, Korea University Sejong Campus, 2511 Sejong-ro, Sejong City, Chungnam 339-700, South Korea. Electronic address: yjhwang@korea.ac.kr.
authors.forename: Youngja
authors.initials: Y
authors.lastname: Park
authors.affiliation: Bio Monitoring Laboratory, Program in Environmental Technology and Policy, Korea University Sejong Campus, 2511 Sejong-ro, Sejong City, Chungnam 339-700, South Korea. Electronic address: ub1905ub@korea.ac.kr.
authors.forename: Sungpyo
authors.initials: S
authors.lastname: Kim
mesh_terms: D000818:Animals; D000890:Anti-Infective Agents / Q000633:toxicity*; D002853:Chromatography, Liquid; D004305:Dose-Response Relationship, Drug; D004784:Environmental Monitoring / Q000379:methods*; D013058:Mass Spectrometry / Q000379:methods; D055442:Metabolome / Q000187:drug effects*; D008954:Models, Biological; D052616:Solid Phase Extraction; D013418:Sulfamethazine / Q000633:toxicity*; D014874:Water Pollutants, Chemical / Q000633:toxicity*; D015027:Zebrafish / Q000378:metabolism*
chemical_list: D000890:Anti-Infective Agents; D014874:Water Pollutants, Chemical; D013418:Sulfamethazine
keywords: Antibiotics; Danio rerio; Environmental metabolomics