SpringerProtocols: Abstract: 11. HPLC Measurement of Aflatoxin B1 and Metabolites in Isolated Rat Hepatocytes

11. HPLC Measurement of Aflatoxin B1 and Metabolites in Isolated Rat Hepatocytes

By: Jennifer Colford²

Abstract

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This chapter describes a high-pressure liquid chromatography (HPLC) method for measuring aflatoxin B₁ and its metabolites isolated from rat hepatocytes. AFB₁, AFQ₁, AFM₁, and AFP₁ concentrations are determined using reverse-phase HPLC. The isocratic mobile phase is 18% dimethylformamide in 0.01% phosphoric acid at a flow rate of 2.0 mL/min at 69°C, with ultraviolet (UV) detection at 360 nm. AFB-GSH conjugate levels are also measured. An isocratic mobile phase consisting of 69.05% H₂O, 30% methanol, and 0.05% acetic acid is used and the sample is eluted at ambient temperature with a flow rate of 1.0 mL/min and UV detection at 365 nm.

Affiliation(s): (2) Food and Research Center, Department of Life Sciences, University of Limerick, Limerick, Ireland

Book Title: Food-Borne Pathogens: Methods and Protocols

Series: Methods in Biotechnology | Volume: 21 | Pub. Date: Oct-06-2005 | Page Range: 125-131 | DOI: 10.1385/1-59259-990-7:125

Subject: Microbiology

Key Words: Aflatoxin - AFB1 - AFM1 - AFP1 - AFQ1 - AFB-GSH conjugate - HPLC - isolated hepatocytes

References

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