Validation of a UHPLC-FLD analytical method for the simultaneous quantification of aflatoxin B1 and ochratoxin a in rat plasma, liver and kidney

Abstract

addresses of all coauthors: Laura-Ana Corcuera: lcorcuer@alumni.unav.es, María Ibáñez-Vea: mivea@alumni.unav.es, Ariane Vettorazzi: avettora@alumni.unav.es, Adela López de Cerain: acerain@unav.es. 2 ABSTRACTA rapid and simple method for the simultaneous quantification of AFB1 and OTA in rat plasma, liver and kidney by UHPLC-FLD has been successfully validated according to the following criteria: selectivity, stability, linearity, precision, accuracy, recovery, robustness and limits of quantification and detection. The extraction method, calibration curves and chromatographic conditions are common for the three matrices. Plasma and homogenized tissue samples (100 µL) were extracted with acetonitrile-formic acid mixture (99:1) (300 µL). Chromatographic separation was performed with a mixture of water and acetonitrile:methanol (50:50), both acidified with 0.5% of formic acid using a gradient profile. The method avoids the use of immunoaffinity columns and allows reduction of sample and solvent volumes as well as toxic wastes. The detection is based on a photochemical reaction which enhances the AFB1 response without affecting the OTA signal before reaching the fluorescent detector. The mycotoxin recovery for each matrix was very efficient, between 93% and 96% for AFB1 and between 94% and 96% for OTA. For both mycotoxins the LOQs were 2 µg/L in plasma and 8 µg/kg in liver and kidney. The method has successfully been applied to rat samples after a single oral administration of a mixture of AFB1 and OTA and it could be a useful tool in toxicokinetic and toxicological studies.

Establishment of an isotope dilution LC-MS/MS method revealing kinetics and distribution of co-occurring mycotoxins in rats

“…Chromatograms of blank plasma (a) and liver (d), blank plasma (b) and liver (e) spiked with AFB1 and T-2 (50 ng mL (Table 2). For both mycotoxins in plasma and tissue homogenates, the LLOD and LLOQ were 0.01 and 0.05 ng mL -1 , respectively (Table 2), which were desirable and obviously lower than those obtained in bibliography via UHPLC-FLD approaches 35 .…”

“…First, the same food can be infected or invaded by different mycotoxigenic molds resulting in the co-occurring AFB1 and T-2 even at high concentrations in many geographic origins under optimum environmental conditions [14][15][16] ; Second, co-occurring mycotoxins can reach humans through the various diets and also, through milk, meat and eggs from livestock and poultry animals fed with different mycotoxins contaminated feedstuffs [17][18][19] . The toxic effects, appearing in consumers exposed to co-occurring mycotoxins, are surely in accordance to the possible interactions of concomitantly occurring mycotoxins, which might be antagonistic, additive, or synergistic on different occasions 20 .…”

Genotoxicity of Aflatoxin B1 and Ochratoxin A after simultaneous application of the in vivo micronucleus and comet assay

“…The extraction procedure and the UHPLC-FLD quantification 7 method was previously set up and validated for these biological samples, and has been performed as described (Corcuera et al, 2011b).…”

Vitamin E (α tocopherol) attenuates toxicity and oxidative stress induced by aflatoxin in rats

“…AFB 1 is nephrotoxic, hepatotoxic, mutagenic, genotoxic, and immunotoxic. [1][2][3][4][5] AFB 1 mediated cell injury may be due to the release of free radicals and these radicals initiate lipid peroxidation (LPO) and a damaging process in biological systems since all cell membranes contain the polyunsaturated fatty acids which are substrates for such a reaction. 6 Kodama et al showed the formation of LPO by AFB 1 .…”