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

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 879; no. 26; pp. 2733 - 2740
• Main Authors: Corcuera, Laura-Ana, Ibáñez-Vea, María, Vettorazzi, Ariane, González-Peñas, Elena, Cerain, Adela López de
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 15.09.2011; Elsevier
• Subjects: Acetonitriles - chemistry; Aflatoxin B1; Aflatoxin B1 - administration & dosage; Aflatoxin B1 - analysis; Aflatoxin B1 - blood; Analysis; Analytical, structural and metabolic biochemistry; Animals; Biological and medical sciences; chromatography; Chromatography, High Pressure Liquid - methods; Female; fluorescence; formic acid; Fundamental and applied biological sciences. Psychology; General pharmacology; Immunoassay; Kidney - chemistry; kidneys; liver; Liver - chemistry; Male; Medical sciences; Ochratoxin A; ochratoxins; Ochratoxins - administration & dosage; Ochratoxins - analysis; Ochratoxins - blood; oral administration; Pharmacology. Drug treatments; Plasma; Rats; Rats, Inbred F344; Reproducibility of Results; solvents; Tissue Distribution; Tissues; toxicity; Toxicity Tests, Acute - methods; UHPLC-FLD; Validation; wastes; UHPLC-FLD; Validation; Aflatoxin B1; Plasma; Tissues; Ochratoxin A; Ultra performance liquid chromatography; Biological fluid; Ochratoxin; Chemical analysis; Rat; Liver; Mycotoxin; Kidney; Blood plasma; Tissue; Quantitative analysis; Digestive system; Rodentia; Toxin; Vertebrata; Mammalia; Urinary system; Animal; Simultaneous measurement; Analytical method
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2011.07.039

A 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.