Development and validation of a rapid multi-biomarker liquid chromatography/tandem mass spectrometry method to assess human exposure to mycotoxins.
BACKGROUND
Mycotoxins regularly occur in food worldwide and pose serious health risks to consumers. Since individuals can be exposed to a variety of these toxic secondary metabolites of fungi at the same time, there is a demand for proper analytical methods to assess human exposure by suitable biomarkers.


METHODS
This study reports on the development of a liquid chromatography/electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) method for the quantitative measurement of 15 mycotoxins and key metabolites in human urine using polarity switching. Deoxynivalenol (DON), DON-3-O-glucuronide, DON-15-O-glucuronide (D15GlcA), de-epoxy DON, nivalenol (NIV), T-2 toxin, HT-2 toxin, zearalenone, zearalenone-14-O-glucuronide, α- and β-zearalenol, fumonisins B(1) and B(2) (FB(1), FB(2)), ochratoxin A (OTA) and aflatoxin M(1) (AFM(1)) were determined without the need for any cleanup using a rapid and simple dilute and shoot approach.


RESULTS
Validation was performed in the range of 0.005-40 µg L(-1) depending on the analyte and expected urinary concentration levels. Apparent recoveries between 78 and 119% and interday precisions of 2-17% relative standard deviation (RSD) were achieved. The applicability of the method was demonstrated by the analysis of urine samples obtained from Cameroon. In naturally contaminated urine samples up to six biomarkers of exposure (AFM(1), DON, D15GlcA, NIV, FB(1), and OTA) were detected simultaneously.


CONCLUSIONS
We conclude that the developed LC/MS/MS method is well suited to quantify multiple mycotoxin biomarkers in human urine down to the sub-ppb range within 18 min and without any prior cleanup. The co-occurrence of several mycotoxins in the investigated samples clearly emphasizes the great potential and importance of this method to assess exposure of humans and animals to naturally occurring mycotoxins.
Center for Analytical Chemistry and Christian Doppler Laboratory for Mycotoxin Metabolism, Department for Agrobiotechnology (IFA-Tulln), University of Natural Resources and Life Sciences, Vienna, Konrad-Lorenz-Str. 20, A-3430 Tulln, Austria.
Benedikt
B
Warth
Michael
M
Sulyok
Philipp
P
Fruhmann
Hannes
H
Mikula
Franz
F
Berthiller
Rainer
R
Schuhmacher
Christian
C
Hametner
Wilfred Angie
WA
Abia
Gerhard
G
Adam
Johannes
J
Fröhlich
Rudolf
R
Krska
D000328:Adult; D015415:Biomarkers / Q000737:chemistry / Q000652:urine; D002163:Cameroon; D002853:Chromatography, Liquid / Q000379:methods* / Q000592:standards; D005506:Food Contamination / Q000032:analysis; D006801:Humans; D016014:Linear Models; D008297:Male; D009183:Mycotoxins / Q000737:chemistry / Q000652:urine*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods* / Q000592:standards
D015415:Biomarkers; D009183:Mycotoxins