Validation of High Performance Liquid Chromatography with Fluorescence Detector Methods for Determination of Aflatoxins in Different Food and Animal Feed Samples
Method validation for quantitative analysis of aflatoxins, AFB1, AFB2,
AFG1 and AFG2 in sorghum, peanut butter, groundnut and animals feed is
presented. Aflatoxins were extracted with a mixture of methanol: acetonitrile:
water (60:30:10) and cleaned with Alfa testIAC
chromatography before analysis by High Performance Liquid Chromatography
coupled with fluorescence detection (HPLC-FLD) by adopting an isocratic
chromatographic system using a mobile phase consisting acetic acid:
acetonitrile: methanol (59:14:27), the separation of the four aflatoxins was
achieved in less than 15 minutes. Calibration curves were linear over the
concentration range from 2 - 18 ng/mL for AFB1 and AFG1, 0.4 - 3.6 ng/mL for AFB2 and AFG2, respectively. The LOD and LOQ in spiked
samples were found to be 0.02 and 0.05 μg/kg for both AFB1 and AFG1, 0.01 and
0.03 μg/kg for both AFB2 and AFG2. The mean recovery values were in range from
84.2% to 96.9% was obtained. Five samples were found to be contaminated with
aflatoxins and the total aflatoxins ranged from 0.02 to3.26
μg/kg were obtained. Nineteen different samples were found to be contaminated
with aflatoxins; the total aflatoxins ranged from 0.27 to10.48
μg/kg were obtained. The highest total aflatoxins value was obtained in animal
feeds.
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