Comparison and improvement of the existing methods for the determination of aflatoxins in human serum by LC-MS/MS

Abstract

Aflatoxins are toxic metabolites of some Aspergillus flavus, A. parasiticus and A. nomius strains that occur in many foods and feeds. Aflatoxin B₁ is the most abundant and toxic member of the family, and it is also the most potent hepatocarcinogen known. In order to estimate the potential human health risk of aflatoxin B₁, it is useful to measure its blood concentration. In this work a rapid liquid chromatography coupled to mass spectrometry (LC-MS/MS) method for the simultaneous determination of aflatoxins B₁, B₂, G₁ and G₂ and its metabolites, aflatoxins M₁ and M₂ in human serum, is proposed. Small serum volumes have been used and the extracts have been obtained without any clean-up procedure. The isolation of the aflatoxins from human serum was achieved using subsequent pH-controlled solvent extractions steps. Recovery rates in the range between 31 and 98% were achieved in the serum, while the limits of detection and quantification for the proposed method were 0.5 ng mL⁻¹ and 1.0 ng mL⁻¹, respectively. The validation of the method has been demonstrated by analysing spiked and naturally contaminated blood serum.