Quantification and confirmation of four aflatoxins using a LC-MS/MS QTRAP system in multiple reaction monitoring, enhanced product ion scan, and MS3 modes.

Abstract

A simple, rapid, and efficient liquid chromatography tandem mass spectrometry (LC-MS/MS) method, operated in electrospray ionization and quadrupole linear ion trap modes, has been developed for the identification and structural characterization of aflatoxins in peanuts and its derivative products or bean sauce. Samples (5 g) were extracted with acetonitrile/water/formic acid (79:20:1, v/v). After centrifugation and dilution, the extracts were separated on a C18 analytical column by gradient elution (acetonitrile/0.2% formic acid) and analyzed by UPLC-MS/MS. External calibration was used for qualification. The developed multiple reaction monitoring-information-dependent acquisition-enhanced product ion method enabled quantification and confirmation of the analytes in a single run. Enhanced product ion mode was used for qualitative analysis, while multiple reaction monitoring mode was used for quantitative analysis. An in-house library was constructed for identification. Calibration curves showed good linearity with correlation coefficients (r) higher than 0.994. Limits of detection were determined to be below 0.26 µg kg-1 for most analytes. The recoveries for those substances were in the acceptable range of 80.2%-119.1%. A new LC-MS3 method was established for further confirmation. One pickled pepper peanut was found to contain aflatoxins B1, B2, and G1 with contents of 90.93, 26.64, and 1.92 µg kg-1, respectively.