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ARS Home » Northeast Area » Wyndmoor, Pennsylvania » Eastern Regional Research Center » Residue Chemistry and Predictive Microbiology Research » Research » Publications at this Location » Publication #318684

Title: Expansion of the scope of AOAC first action method 2012.25 - single-laboratory validation of triphenylmethane dye and leuco metabolite analysis in shrimp, tilapia, catfish, and salmon by LC-MS/MS

item ANDERSEN, WENDY - Us Food & Drug Administration (FDA)
item CASEY, CHRISTINE - Us Food & Drug Administration (FDA)
item Schneider, Marilyn
item TURNIPSEED, SHERRI - Us Food & Drug Administration (FDA)

Submitted to: Journal of AOAC International
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 12/1/2014
Publication Date: 5/1/2015
Publication URL:
Citation: Andersen, W.C., Casey, C.R., Schneider, M.J., Turnipseed, S.B. 2015. Expansion of the scope of AOAC First Action Method 2012.25 - single-laboratory validation of triphenylmethane dye and leuco metabolite analysis in shrimp, tilapia, catfish, and salmon by LC-MS/MS. Journal of AOAC International. DOI: 10.5740/jaoacint.14-264.

Interpretive Summary: Analytical methods used for monitoring of chemical contaminants in food must be accurate because the results are used in regulatory enforcement actions and trade disputes. The gold standard for method validation involves interlaboratory trials using shared samples, such as Collaborative Studies in the AOAC International Official Methods Program. This study demonstrated that the method and samples were ready for evaluation and use in an AOAC International Collaborative Study, which was successfully conducted afterwards among many labs worldwide. Regulatory actions and other decisions can be made with more confidence based on the results of this method, which can involve millions of dollars of seafood product.

Technical Abstract: Prior to conducting a collaborative study of AOAC First Action 2012.25 LC-MS/MS analytical method for the determination of residues of three triphenylmethane dyes (malachite green, crystal violet, and brilliant green) and their metabolites (leucomalachite green and leucocrystal violet) in seafood, a single-laboratory validation of method 2012.25 was performed to expand the scope of the method to other seafood matrixes including salmon, catfish, tilapia, and shrimp. The validation included the analysis of fortified and incurred residues over multiple weeks to assess analyte stability in matrix at –80 degree C, a comparison of calibration methods over the range 0.25 to 4 ug/kg, study of matrix effects for analyte quantification, and qualitative identification of targeted analytes. Method accuracy ranged from 88 to 112% with 13% RSD or less for samples fortified at 0.5, 1.0, and 2.0 ug/kg. Analyte identification and determination limits were determined by procedures recommended both by the U.S. Food and Drug Administration and the European Commission. Method detection limits and decision limits ranged from 0.05 to 0.24 ug/kg and 0.08 to 0.54 ug/kg, respectively. AOAC First Action Method 2012.25 with an extracted matrix calibration curve and internal standard correction is suitable for the determination of triphenylmethane dyes and leuco metabolites in salmon, catfish, tilapia, and shrimp by LC-MS/MS at a residue determination level of 0.5 ug/kg or below.