Analysis of malachite green and metabolites in fish using liquid chromatography atmospheric pressure chemical ionization mass spectrometry

Rapid Commun Mass Spectrom. 1998;12(21):1625-34. doi: 10.1002/(SICI)1097-0231(19981115)12:21<1625::AID-RCM373>3.0.CO;2-I.


Malachite green (MG), a traditional agent used in aquaculture, is structurally related to other carcinogenic triphenylmethane dyes. Although MG is not approved for use in aquaculture, its low cost and high efficacy make illicit use likely. We developed sensitive and specific methods for determination of MG and its principal metabolite, leucoMG (LMG), in edible fish tissues using isotope dilution liquid chromatography atmosphere pressure chemical ionization mass spectrometry. MG and LMG concentrations were measured in filets from catfish treated with MG under putative use conditions (ca. 250 and 1000 ppb, respectively) and from commercial trout samples (0-3 and 0-96 ppb, respectively). Concentrations of LMG in edible fish tissues always exceeded those of MG. A rapid cone voltage switching acquisition procedure was used to simultaneously produce molecular ions for quantification and diagnostic fragment ions for confirmation of MG and metabolites. The accurate and precise agreement between diagnostic ion intensity ratios produced by LMG in authentic standards and incurred fish samples was used to unambiguously confirm the presence of LMG in edible fish tissue. This suggested the validity of using LMG as a marker residue for regulatory determination of MG misuse. Additional metabolites derived from oxidative metabolism of MG or LMG (demethylation and N-oxygenation) were identified in catfish and trout filets, including a primary arylamine which is structurally related to known carcinogens. The ability to simultaneously quantify residues of MG and LMG, and to confirm the chemical structure of a marker residue by using LC/MS, suggests that this procedure may be useful in monitoring the food supply for the unauthorized use of MG in aquaculture.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Aniline Compounds / urine
  • Animals
  • Biotransformation
  • Catfishes / metabolism*
  • Chromatography, Liquid
  • Gentian Violet / urine
  • Indicators and Reagents
  • Mass Spectrometry
  • Meat / analysis
  • Muscle, Skeletal / chemistry
  • Rosaniline Dyes / blood
  • Rosaniline Dyes / pharmacokinetics*
  • Trout / metabolism*


  • Aniline Compounds
  • Indicators and Reagents
  • Rosaniline Dyes
  • malachite green
  • leucomalachite green
  • Gentian Violet