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Validation of an LC-MS/MS method for malachite green (MG), leucomalachite green (LMG), crystal violet (CV) and leucocrystal violet (LCV) residues in fish and shrimp

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A quantitative liquid chromatography–tandem mass spectrometry (LC-MS/MS) method for the simultaneous analyses of malachite green (MG), crystal violet (CV) and its major metabolites, leucomalachite green (LMG) and leucocrystal violet (LCV) residues in fish and shrimp samples has been validated. Fish and shrimp samples were extracted with citrate buffer/acetonitrile, and the extracts were purified on strong cation-exchange (SCX) solid-phase extraction (SPE) cartridge. After conversion of LMG into MG using a post column oxidation reactor containing lead (IV) oxide (PbO2), the effluents were analysed. Residues were analysed using positive-ion electrospray ionisation (ESI). Identification and quantification of analytes were based on the ion transitions monitored by multiple reaction monitoring (MRM). Validation of the method was carried out in accordance with the Decision 2002/657/EC, which establishes criteria and procedures for the validation of methods. The following parameters were determined: decision limit (CCα), detection capability (CCβ), linearity, accuracy, precision, selectivity, specificity and matrix effect. The decision limits (CCα) for MG, LMG, CV and LCV were 0.164, 0.161, 0.248 and 0.860 µg kg–1. The respective detection capabilities (CCβ) were 0.222, 0.218, 0.355 and 1.162 µg kg–1. Typical recoveries (intermediate precision) in shrimp, for MG, CV, LMG and LCV for 2.0 µg kg–1 level fortified samples using the optimised procedure were in the range 69%, 97%, 80.3% and 71.8%, respectively. The findings demonstrate the suitability of the method to detect simultaneously MG, CV and its metabolite (LMG and LCV) in fish and shrimp.
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Keywords: analysis; dyes; fish; leucomalachite green; malachite green; method validation; shrimp

Document Type: Research Article

Affiliations: 1: Bolsista CNPq,Laboratório Nacional Agropecuário (LANAGRO), Pedro Leopoldo, Brazil 2: Laboratório de Resíduos de Medicamento Veterinários,Laboratório Nacional Agropecuário (LANAGRO), Pedro Leopoldo, Brazil

Publication date: April 1, 2012

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