title: Determination of malachite green, crystal violet and their leuco-metabolites in fish by HPLC-VIS detection after immunoaffinity column clean-up.
abstract: A high performance liquid chromatography method with visible detection (HPLC-VIS) for the determination of malachite green (MG), crystal violet (CV), leucomalachite green (LMG), and leucocrystal violet (LCV) in fish has been developed after clean-up through an immunoaffinity column (IAC). Residues were simultaneously extracted from fish muscle with acetonitrile and ammonium acetate buffer. The leuco-forms, LMG and LCV, were oxidized quantitatively to the chromic CV and MG by reaction with 2,3-dichloro-5,6-dicyano-1,4-benzoquinone. Extracts were then purified on an IAC which prepared by immobilizing the anti-MG-CV antibodies by the sol-gel method. Finally, the eluents were analyzed by HPLC-VIS. The limits of detection were 0.15, 0.1, 0.18 and 0.14ng/g for MG, CV, LMG and LCV, respectively. The average recoveries in samples fortified with MG, CV, LMG and LCV over the range 0.5-10ng/g were from 71.6% to 96.8% with RSDs of 5.1-12.3% (n=6). This novel method was confirmed by liquid chromatography-tandem mass spectrometry with electrospray interface in positive mode using multiple reaction monitoring.
authors.affiliation: Department of Chemistry and Chemical Engineering, University of South China, Hunan, Hengyang 421001, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Xie
authors.forename: Tao
authors.initials: T
authors.lastname: Peng
authors.forename: Dong-Dong
authors.initials: DD
authors.lastname: Chen
authors.forename: Qing-Jie
authors.initials: QJ
authors.lastname: Zhang
authors.forename: Guo-Min
authors.initials: GM
authors.lastname: Wang
authors.forename: Xiong
authors.initials: X
authors.lastname: Wang
authors.forename: Qi
authors.initials: Q
authors.lastname: Guo
authors.forename: Fan
authors.initials: F
authors.lastname: Jiang
authors.forename: Dan
authors.initials: D
authors.lastname: Chen
authors.forename: Jian
authors.initials: J
authors.lastname: Deng
mesh_terms: D000097:Acetonitriles / Q000737:chemistry; D000818:Animals; D002846:Chromatography, Affinity / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005399:Fishes; D005840:Gentian Violet / Q000032:analysis*; D057230:Limit of Detection; D009132:Muscles / Q000737:chemistry; D010084:Oxidation-Reduction; D015203:Reproducibility of Results; D012394:Rosaniline Dyes / Q000032:analysis*
chemical_list: D000097:Acetonitriles; D012394:Rosaniline Dyes; C005095:malachite green; C015320:leucomalachite green; D005840:Gentian Violet; C032159:acetonitrile