<p>Aflatoxins (AFs) pose a significant health risk due to their toxicity and carcinogenicity, with spices being particularly vulnerable to contamination. This study develops and validates a robust LC-ESI-MS/MS method for the simultaneous quantification of AFB1, AFB2, AFG1, and AFG2 in Moroccan saffron and fennel (&lt; 15% moisture). The optimized sample preparation combines a modified QuEChERS extraction (10 mL ACN/water, 50:50 v/v, 1% acetic acid) with µ-dSPE cleanup (200 mg MgSO₄, 100 mg C18), reducing matrix effects by 40% compared to conventional protocols. Chromatographic separation achieved baseline resolution in 6 min using a C18 column (150 × 3 mm, 2.7 µm) with ESI+ in MRM mode. The method demonstrated excellent linearity (1–20 µg/kg, <i>R</i>² &gt; 0.995), low LODs (0.26–0.37 µg/kg), and LOQs (0.53–0.74 µg/kg). Recoveries (<i>n</i> = 6) ranged from 80 to 118% (1–20 µg/kg) with RSDs &lt; 15%. Expanded uncertainty (<i>k</i> = 2) was &lt; 50%, complying with EU Regulation 2023/915. Robustness and applicability were further confirmed through interlaboratory proficiency testing, with accurate and reproducible results. Notably, this is the first report of AF contamination levels in Moroccan saffron and fennel, revealing concentrations below EU limits (5 µg/kg AFB1, 10 µg/kg total AFs). The method was successfully applied to Moroccan saffron and fennel seeds, revealing no detectable aflatoxin contamination. The method’s throughput (6 min/run) and cost-efficiency make it ideal for routine food safety monitoring in resource-limited settings.</p> Graphical Abstract <p></p>