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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 liquid
chromatography – tandem quadrupole mass spectrometry (AQUITY
UPLC–TQD) was developed and validated for the determination and
confirmation of flubendazole in Galda shrimp (Macrobrachium
rosenbergii) and Katla (Katla katla) fish. The UPLC separation was
achieved on a AQUITY UPLC BEH C18 Column (2.1x50mm; 1.7μm)
with gradient elution using a mobile phase of 0.1% formic acid in water
and 0.1% formic acid in acetonitrile. Mass spectral acquisition was
done using electrospray ionization in the positive ion mode applying
multiple reaction monitoring (MRM) of two diagnostic transition
reactions for flubendazole (m/z 314.077 282.022 and m/z 314.077 122.899) and for
Flubendazole_d3 (m/z 317.077 282.028). Shrimp samples were extracted with ethyl acetate
and evaporated to dryness. The method validation was carried out according to the criteria of
Commission decision 2002/657/EC. The calibration curve showed a good linearity in the
concentration range from 1 to 20 ng/g with the correlation coefficient of >0.996. In case of
shrimp samples the decision limit (CCα) and the detection capability (CCβ) were 6.0 μg/kg
and 7.1 μg/kg respectively and that for fish sample CCα value was 6.17 μg/kg and CCβ value
was 7.45 μg/kg for flubendazole. The mean recoveries were in the range of 90–108% for
shrimp and that of 90-105% for fish. The precision of the method, expressed as RSD values
for 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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