Development of a liquid chromatography/tandem mass spectrometry method for the quantification of fumonisin B₁, B₂ and B₃ in cornflakes

Abstract

A liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for the determination of fumonisin B1 (FB1), B2 (FB2) and B3 (FB3) in cornflakes is described. During method development, special attention was paid to the selection of a suitable internal standard (IS) in order to offer a good alternative for deuterated FB1. In this respect, the C12-sphinganine analogue (2S,3R)-2-aminododecane-1,3-diol was chosen because of its structural similarity to the fumonisin backbone and its chromatographic elution between the target analytes. For the extraction of the fumonisins from the cornflakes matrix, MeOH/H2O (adjusted to pH 4 with 0.1 M HCl; 70:30, v/v), ACN/MeOH/H(2)O (25:25:50, v/v/v) and acidified ACN/MeOH/H2O (25:25:50, v/v/v; pH 4) were evaluated. Preference was given to acidified MeOH/H2O (70:30, v/v) with mean recoveries (n=12) for FB1, FB2 and FB3 of, respectively, 84+/-10, 78+/-7 and 85+/-9%. Cleanup was performed using immunoaffinity columns (FumoniTest, VICAM). The chromatography was performed under isocratic conditions at a flow of 0.3 mL min-1 with a mobile phase consisting of ACN/H2O (60:40, v/v) containing 0.3% formic acid. The mass spectrometer was operated in the positive electrospray ionization (ESI+) mode using multiple reaction monitoring (MRM). An intralaboratory validation was conducted with fortified samples determining limits of detection (LOD), limits of quantification (LOQ), precision, trueness, specificity and measurement uncertainty. The LOD concentrations for FB1, FB2 and FB3 were 20, 7.5 and 12.5 microg/kg. The LOQs were 40 microg/kg for FB1, 15 microg/kg for FB2 and 25 microg/kg for FB3. The coefficients of variation (CVs) under repeatability conditions varied from 11 to 13% for FB1, from 9 to 14% for FB2 and from 7 to 10% for FB3. Under within-laboratory reproducibility conditions, the CVs ranged from 12 to 17% for FB1, from 9 to 16% for FB2 and from 7 to 13% for FB3. The percent bias for FB1 varied from -12 to -10%, while for FB2 and FB3 bias ranged, respectively, from -4 to -2% and from -12 to -5%. The expanded measurement uncertainties for FB1, FB2 and FB3 were, respectively, 19, 18 and 22%.

Immunoaffinity column clean-up techniques in food analysis: A review

“…It can also be seen from Table 3 that IACs have been successfully used by laboratories, where only TLC is available [44,45,47], and the improved clean-up means that the lack of separation on the TLC plate becomes less disadvantageous. At the other end of the spectrum IAC clean-up has been used with sophisticated chromatographic end-determinations being used in LC/MS and LC/MS/MS [107,108].…”

Simultaneous determination of aflatoxins, ochratoxin A andFusariumtoxins in maize by liquid chromatography/tandem mass spectrometry after multitoxin immunoaffinity cleanup

“…In this regard, the most critical step is the extraction that should allow good recoveries for all mycotoxins under investigation in the specific food matrix. Several authors have reported that high methanol or acetonitrile concentrations (≥75%) in the extraction solvent are suitable for the extraction of most mycotoxins, such as trichothecenes, ZEA, OTA, and AFs,6–9 whereas high extraction efficiencies for FBs can be obtained by increasing the water fraction and/or lowering the pH of the extraction mixture 10–12…”

Development of a multi‐mycotoxin liquid chromatography/tandem mass spectrometry method for sweet pepper analysis

“…Until now no isotopically labeled internal standards have been used in multi‐mycotoxin LC/MS/MS methods. Another approach is the use of structural analogues such as the sphinganine analogue (2 S ,3 R )‐2‐aminododecane‐1,3‐diol to determine FB 1 , FB 2 and FB 3 in corn‐flakes 1, 22. The use of verrucarol was reported to determine Fusarium toxins 23.…”