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Abstract

DEVELOPMENT AND VALIDATION OF A CONFIRMATORY METHOD FOR FLUBENDAZOLE IN FISH AND SHRIMP MATRICES BY LC-MS/MS SYSTEM

Md. Ashraful Alam*, Akter Mst Yeasmin, Talukdar Muhammad Waliullah, Saleh
Ahmed, Md. Serajul Islam and Md. Manik Mia

Abstract

A liquid chromatographic method using ultra performance liquidchromatography – tandem quadrupole mass spectrometry (AQUITYUPLC–TQD) was developed and validated for the determination andconfirmation of flubendazole in Galda shrimp (Macrobrachiumrosenbergii) and Katla (Katla katla) fish. The UPLC separation wasachieved on a AQUITY UPLC BEH C18 Column (2.1x50mm; 1.7μm)with gradient elution using a mobile phase of 0.1% formic acid in waterand 0.1% formic acid in acetonitrile. Mass spectral acquisition wasdone using electrospray ionization in the positive ion mode applyingmultiple reaction monitoring (MRM) of two diagnostic transitionreactions for flubendazole (m/z 314.077 282.022 and m/z 314.077 122.899) and forFlubendazole_d3 (m/z 317.077 282.028). Shrimp samples were extracted with ethyl acetateand evaporated to dryness. The method validation was carried out according to the criteria ofCommission decision 2002/657/EC. The calibration curve showed a good linearity in theconcentration range from 1 to 20 ng/g with the correlation coefficient of >0.996. In case ofshrimp samples the decision limit (CCα) and the detection capability (CCβ) were 6.0 μg/kgand 7.1 μg/kg respectively and that for fish sample CCα value was 6.17 μg/kg and CCβ valuewas 7.45 μg/kg for flubendazole. The mean recoveries were in the range of 90–108% forshrimp and that of 90-105% for fish. The precision of the method, expressed as RSD valuesfor the within laboratory reproducibility at the three levels of fortification (2.5, 5.0 and 7.5)μg/kg was less than 15%

Keywords: UPLC-TQD, Mass Spectrometry, Flubendazole, Shrimp, Fish and Validation.


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