Journal of AOAC International

Liquid chromatographic method for nicarbazin in broiler feeds and premixtures: development, validation, and interlaboratory study.

PMID 15675436


A reversed-phase liquid chromatography method for nicarbazin in broiler feeds and premixtures was developed, validated, and interlaboratory studied. The extraction solvent was an acetonitrile-methanol (1 + 1) mixture. For feedingstuffs, water was also added. The 4,4'-dinitrocarbanilide moiety of nicarbazin was detected at a wavelength of 350 nm. Recovery was > or =87%. At 20 mg/kg, the repeatability was 0.7% and the within-laboratory reproducibility was 2.7%. The limit of determination was <20 mg/kg. Other feed additives did not interfere in the assay that proved to be applicable to broiler feeds from different European Union countries. In an interlaboratory study, 4 positive broiler feeds, 1 blank pig feed, and 1 broiler premixture were analyzed by 19 laboratories using the method developed in this study. The relative standard deviation for repeatability (RSDr) of the feedingstuffs (20-240 mg/kg) varied between 2.6 and 10.2%. The HORRAT ranged between 0.70 and 1.22. Recoveries were 91-108%. Three laboratories detected small signals in the blind blank samples, ranging from 0.4 to 2 mg/kg. For the premixture, acceptable results for reproducibility could only be obtained after the sample weight and volume of extraction had been doubled. To avoid excessive dilution of the extracts, the range of the calibration curve had also been doubled. With this modified method, the RSDr was 5.7% and the HORRAT was 1.95 (10 laboratories).

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C13H10N4O5 · C6H8N2O
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C13H10N4O5 · C6H8N2O