PRiME pass-through cleanup for the fast determination of aflatoxins in human serum by using LC-MS/MS

Abstract

A novel, simple and sensitive method based on the use of a PRiME (process, robustness, improvements, matrix effects, ease of use) pass-through cleanup procedure combined with ultra-fast liquid chromatography-tandem quadrupole mass spectrometry (UFLC-MS/MS) for the determination of 4 AFs, i.e., AFB₁, AFB₂, AFG₁ and AFG₂ in human serum, was developed and validated. The effective PRiME pass-through cleanup procedure using the Oasis PRiME HLB cartridge was optimized for the extraction of AFB₁, AFB₂, AFG₁ and AFG₂ in human serum. The obtained results demonstrated the higher extraction capacity of the PRiME pass-through cleanup procedure with recoveries between 92.1% and 102%. The limits of detection (LODs) for AFB₁, AFB₂, AFG₁ and AFG₂ were 0.048 μg L⁻¹, 0.069 μg L⁻¹, 0.043 μg L⁻¹ and 0.14 μg L⁻¹, respectively. Validation results on linearity, specificity, accuracy and precision, as well as on application to the analysis of AFs in 10 liver cancer volunteers demonstrated the applicability to clinical studies.