A miniaturized QuEChERS-DLLME method for simultaneous determination of acetamide, acrylamide, and glycidamide in infant formula using GC–MS

• Simultaneous GC–MS determination of acrylamide, glycidamide and acetamide. • µ-QuEChERS simplifies infant formula's defatting and protein precipitation. • µ-QuEChERS’ organic waste used as dispersant phase in DLLME. • In-situ xanthydrol derivatization enhance sensitivity in a reduced time. • Sustainable approach for infant formula analysis and food safety. Infant formula is the only food product suitable for infants during the first months of life. Therefore, evaluating food contaminants is essential for ensuring infant food safety. We developed a miniaturized QuEChERS (µ-QuEChERS) method combined with Dispersive Liquid-Liquid Microextraction (DLLME) to detect acrylamide, glycidamide, and acetamide in IF simultaneously. Several parameters were optimized, including sample size, extraction solvent, clean-up sorbent type, and amount, among others, using an asymmetrical factorial design. The method also incorporated xanthydrol derivatization under light-protected, acidic conditions within the DLLME process, improving selectivity and enrichment. The µ-QuEChERS-DLLME method was validated according to FDA guidelines, demonstrating high selectivity, specificity, and excellent linearity ( r ² ≥ 0.9995). Recovery rates ranged from 91.0 % to 110.1 %, with precision (RSD ≤ 9.1 %). The method's sustainability was evaluated using the AGREEprep tool. It was applied to analyze acrylamide, glycidamide, and acetamide in 8 infant formula samples. Acetamide was quantifiable in one sample (230 ng/g), while acrylamide levels ranged from below the quantification limit to 50 ng/g, consistent with previous studies. This demonstrates the method's robustness for amide analysis in infant formula.