ZrO₂-Assisted QuEChERS-UHPLC-MS/MS for Simultaneous Determination of Four Aflatoxins in Cereals and Soybean Matrices

Highly sensitive methods for trace-level aflatoxin determination are indispensable for cereal food safety and public health protection. This study developed a ZrO₂-assisted QuEChERS-UHPLC-MS/MS method for the simultaneous determination of AFB₁, AFB₂, AFG₁, and AFG₂ in maize, wheat, rice, and soybean. Systematic optimization identified acetonitrile as the optimal extraction solvent and 10 mg ZrO₂ in combination with PSA, C₁₈, and GCB as the optimal cleanup formulation, providing recoveries of 107.33-111.60%. Chromatographic baseline separation was achieved within 8.0 min using a moderate gradient program. The method exhibited excellent linearity (R² > 0.999) with LODs of 0.15-0.25 µg/kg and LOQs of 0.50-0.75 µg/kg. Negligible matrix effects (0.85-1.02) validated the efficacy of ZrO₂-assisted cleanup in eliminating co-extractive interferences in maize. Satisfactory accuracy (recoveries of 86.66-111.04%) and precision (RSDs < 14%) were obtained across all matrices. The method demonstrated consistent performance across diverse cereal and soybean matrices, fulfilling international regulatory requirements for routine aflatoxin monitoring in agricultural commodities.