title: A selective biomarker for confirming nitrofurazone residues in crab and shrimp using ultra-performance liquid chromatography-tandem mass spectrometry.
abstract: Reliably detecting nitrofurazone (NFZ) residues in farmed crab and shrimp was previously hindered by lack of appropriately specific analytical methodology. Parent NFZ rapidly breaks down in meat, and the commonly used side-chain metabolite, semicarbazide (SEM), is non-specific as it occurs naturally in crustacean shell often leading to 'false positive' detections in meat. Using 5-nitro-2-furaldehyde (NF) as marker metabolite, following pre-column derivatization with 2,4-dinitrophenylhydrazine (DNPH), ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) analysis in negative electrospray ionization mode enabled confirmation of NFZ residues in deliberately treated whole crab, crab meat and shrimp meat, with a limit of detection (LOD) and limit of quantification (LOQ) below 1 ng g(-1). Meanwhile, the derivatives of DNPH-NF were synthesized for the first time, purified by preparative liquid chromatography and structure characterized with nuclear magnetic resonance spectroscopy ((1)H-NMR). The purity of derivative was checked by ultra-performance liquid chromatography-tunable ultraviolet (UPLC-TUV), and the contents were beyond 99.9%. For comparison purposes, crustacean samples were analysed using both NF and SEM marker metabolites. NFZ treatment was revealed by both NF and SEM marker metabolites, but untreated crab also showed measurable levels of SEM which could potentially be misinterpreted as evidence of illegal NFZ use.
authors.affiliation: Key Lab of Mariculture and Enhancement, Marine Fisheries Research Institute of Zhejiang Province, Zhoushan, 316000, China.|Marine and Fisheries Research Institute, Zhejiang Ocean University, Zhoushan, Zhejiang, 316000, China.
authors.forename: Shuai
authors.initials: S
authors.lastname: Zhang
authors.affiliation: Key Lab of Mariculture and Enhancement, Marine Fisheries Research Institute of Zhejiang Province, Zhoushan, 316000, China.
authors.forename: Yuanming
authors.initials: Y
authors.lastname: Guo
authors.affiliation: Key Lab of Mariculture and Enhancement, Marine Fisheries Research Institute of Zhejiang Province, Zhoushan, 316000, China.
authors.forename: Zhongyong
authors.initials: Z
authors.lastname: Yan
authors.affiliation: Key Lab of Mariculture and Enhancement, Marine Fisheries Research Institute of Zhejiang Province, Zhoushan, 316000, China.
authors.forename: Xiumei
authors.initials: X
authors.lastname: Sun
authors.affiliation: Key Lab of Mariculture and Enhancement, Marine Fisheries Research Institute of Zhejiang Province, Zhoushan, 316000, China. zhangxj_hys@163.com.
authors.forename: Xiaojun
authors.initials: X
authors.lastname: Zhang
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000737:chemistry* / Q000378:metabolism; D003386:Brachyura / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004350:Drug Residues / Q000737:chemistry* / Q000378:metabolism; D005506:Food Contamination / Q000032:analysis; D005662:Furaldehyde / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism; D015394:Molecular Structure; D009583:Nitrofurazone / Q000737:chemistry* / Q000378:metabolism; D033541:Palaemonidae / Q000737:chemistry*; D012663:Semicarbazides / Q000737:chemistry / Q000378:metabolism; D012758:Shellfish / Q000032:analysis*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D015415:Biomarkers; D012663:Semicarbazides; C010059:carbamylhydrazine; D005662:Furaldehyde; D009583:Nitrofurazone; C052590:5-nitro-2-furaldehyde
keywords: 2,4-Dinitrophenylhydrazine (DNPH); 5-Nitro-2-furaldehyde (NF); Nitrofurazone metabolites; Semicarbazide (SEM); UPLC–MS/MS