**3. Conclusions**

The in-house validation data presented here show that the improved aflatoxin test procedure is suitable for estimation of aflatoxin contamination levels in chicken feed. The optimal aflatoxin B1 extraction conditions are wet milling (water slurring 25 g feed in 37.5 mL water), solid–liquid extraction 1.3 g slurry (0.52 matrix: 0.78 water, *w*/*w*) with acetonitrile:water (80:20, *v*/*v*) at dilution rate of slurry/acetonitrile solvent (1:66.5, *w*/*v*) followed by extraction back to aqueous phase at dilution rate of extract/phosphate buffered saline tween 20 (1:6.5, *v*/*v*). The improved aflatoxin test procedure is an accurate, precise, stable, reliable and cost-effective tool (in terms of time and resources) for surveillance of dietary aflatoxin. The improved aflatoxin test procedure described herein is especially suitable for laboratories that may not have access to automated sample-splitting equipment.
