Optimization and validation of a liquid chromatography/tandem mass spectrometry (LC-MS/MS) method for the determination of aflatoxins in maize

Abstract

A LC-MS/MS method has been optimized and validated for the determination of aflatoxins (AFB1, AFB2, AFG1 and AFG2) in maize. Extraction was performed using a modified QuEChERS method with little sample preparation without the need for purification procedure. Determination was performed by high pressure liquid chromatography (HPLC) coupled to tandem mass spectrometry (MS/MS). The acquisition was performed using MassHunter software in Multiple Reaction Monitoring (MRM) mode in positive polarity. Different mobile phases were tested to control the degree of the ionization and good performances were obtained for methanol/water with 5 mM ammonium acetate. MRM experiments were optimized for each aflatoxin in order to generate sensitive transitions. Linearity was demonstrated for the aflatoxins in the range 0.225–1.25 μg/L. Limits of detection (LOD) (0.11 and 0.36 μg/Kg) and limits of quantification (LOQ) (0.36–1.19 μg/Kg) of the aflatoxins are below the maximum permitted levels set by the European Union (EU). Aflatoxins have acceptable recoveries using QuEChERS method in the acceptable range of 50–120% for levels below 1 μg/Kg. Satisfactory recoveries were also obtained in the acceptable range of 70–110% for levels between 1 and 10 μg/Kg except for AFB2. Relative standard deviation (RSD) of recoveries for the intra-day precision and inter-day precision were below 11 %. Selectivity of the method was tested and no spectral interferences were observed in the appropriate retention times. The main advantage of the proposed method is its ease of use and requires a smaller solvent consumption that reduces the time and cost of the analysis.