Optimization And Research On Determination Method Of Malachite Green In Aquatic Products

Malachite green(MG) belong to triphenylmaethane compound, had been used would wide as antiparasitic and fungicide to prevent and treat fish diseases in aquaculture. However, due to their mutagenic, teratogenic and carcinogenic natures, they had been prohibited to be usded in fishery in many countries. Ministry of Agriculture of China had completely banned the use of MG in food animal production in May 2002.It couldn’t be detected in all of the animal food. Nevertheless, MG is illegal used because of its excellent antibacterial with low cost. So the rapid, sensitive and accurate determination methods for the banned veterinary like malachite green in production and distribution are very necessary.In this article, we depend on existing national standards, establish A UPLC-ESI-MS/MS method for rapid determination of malachite green (MG) and its metabolite leucomalachite green(LMG) in aquatic fingerlings with quantification by isotope-labeled internal standard method. UPLC and MS/MS analytical conditions were examined and optimized by a series of experiments. The compound was oxidated by buffered salt system and acid Al2O3, extracted by acetonitrile and dichloromethane, concentrated liquid was purified on a neutral Al2O3 solid phase extraction cartridge. All eluates were collected and evaporated to dryness. The residue was re-dissolved in 5m mol/L ammonium acetate with 0.1% Formic acid and Methanol (1:1,V/V), sonficated and filtered prior to UPLC analysis. Malachite green and leucomalachite green showed good linearity over the range of 0.5 to 20.0 ng/mL with correlation coefficient of 0.99991 and 0.99995, through the 150 samples in 10 different types of seafood products spiked recovery experiments, the average recoveries at three spiked levels (1.0μg/kg,2.0μg/kg and 4.0μg/kg) ranged from 81.9% to 102.3% with relative standard deviations from 2.28%to 8.74%.Repeated the test about the limits of detection in National standard, the limits of this method were 0.30μg/kg for malachite green and leucomalachite green.In addition, the method showed advantages of simplicity, accuracy, sensitivity and coverage, Compare the method with Literature and standard, it has made some improvements in the before treatment, it could be applied for simultaneous determination of malachite green and leucomalachite green in FuJian seafood product, and provided a reliable supervision for aquaculture and circulation in FuJian.