Abstract
<title>Abstract</title> <p>Acidified methanol is widely used for lipid residue analysis in archaeological pottery, but protocols vary among laboratories, and the effects of key parameters on lipid recovery remain unclear. This study evaluated sulfuric acid concentration, solid-liquid ratio, extraction cycles, and water bath time, using recovery to assess extraction efficiency. Results showed that a 5% sulfuric acid concentration, a 1:5 solid-liquid ratio, and two extraction cycles yielded the optimal recovery, while shortening the water bath time improved analytical efficiency without reducing recovery. Subsequently, the optimized protocol was applied to selected real archaeological pottery samples, validating its applicability to the real pottery matrix and its ability to detect representative lipid residues. Statistical analysis indicated that the optimized parameters significantly affected lipid recovery, and interactions existed among key variables. The resulting analytical procedure is robust, reliable, and highly reproducible, suitable for promoting standardization of archaeological lipid residue analysis across laboratories.</p>