Abstract

This paper describes a reserved-phase high-performance liquid chromatographic (HPLC) method for detecting malachite green (MG) and leuco-malachite green (LMG) using an isocratic toxic organic solvent/reagent-free mobile phase. Chromatographic separations were performed an Inertsil® WP300 C4 with 0.02 mol/L octane sulfonic acid–ethanol mobile phase and a photodiode-array detector. The total run time was <5 min. The system suitability was well within the international acceptance criteria. A harmless method for simultaneously detecting MG and LMG was developed and may be further applied to the quantification in foods.

Highlights

  • Malachite green (MG) is a basic organic bluish green pigment and is used in the treatment of infectious diseases of ornamental fishes, such as Saprolegniasis caused by a fungus belonging to the genus Saprolegnia and ichthyophthirius disease [1,2]

  • Numerous reports of MG misuse in aquaculture have been reported in the US and internationally [4], and residues of MG and its metabolite, leuco-malachite green (LMG) are the most prohibited compounds found in aquaculture products [5,6]

  • Several high-performance liquid chromatographic (HPLC) methods have been developed for the monitoring MG and LMG with different detection modes [7,8,9,10,11,12,13], these methods have crucial drawbacks

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Summary

Introduction

Malachite green (MG) is a basic organic bluish green pigment and is used in the treatment of infectious diseases of ornamental fishes, such as Saprolegniasis caused by a fungus belonging to the genus Saprolegnia and ichthyophthirius disease [1,2]. All of the methods consume large quantities of toxic organic solvents, acetonitrile and/or methanol [14], in the mobile phases. Risks associated with these solvents extend beyond direct implications for the health of humans and wildlife to affecting our environment and the ecosystem in which we all reside. The availability of the LC-MS/MS system is limited to industrial nations because these are hugely expensive, and the methodologies are complex and specific These are unavailable in a lot of laboratories for routine analysis, in developing countries. As the first examination problem in the establishment of an international harmonized method for the residue monitoring of MG and LMG, this paper describes isocratic mobile phase HPLC conditions to detect MG and LMG without toxic solvent/reagent consumption

Chemicals and Reagents
Equipments
Operating Conditions
Preparation of Stock Standards and Working Mixed Solutions
System Suitability Test
Optimum HPLC Conditions
Main Validation Data
Robustness
Conclusions

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