Acrylamide Determination in Infant Formulas: A New Extraction Method

Infant formulas are specialized foods designed for babies and toddlers who cannot be exclusively breastfed. However, acrylamide (AA) may form during the thermal processing involved in their production. Although chromatographic techniques offer high sensitivity and detection capability for AA analysis, their application remains limited due to the complexity of diverse food matrices, high operating costs, time requirements, and environmental concerns. A new validated liquid chromatography-mass spectrometry (LC-MS) protocol for AA detection in infant formula was developed using sequential hydration, acetonitrile (ACN) precipitation, and dual-sorbent clean-up, which minimized matrix effects and ensured clarity and high reproducibility. The validated method demonstrated excellent linearity (R² = 0.9985, solvent-based; 0.9903, matrix-based), a pronounced matrix effect (-67%), satisfactory sensitivity (limit of detection, LOD: 10 µg/kg; limit of quantification, LOQ: 20 µg/kg), and consistent recovery (82-99%) with less than 15% variation. AA analysis was performed on 31 infant formula samples. The highest individual AA level (268.2 µg/kg) was detected in an amino acid-based formula intended for infants under one year of age while the highest mean concentration was found in cereal-based samples (188.1 ± 100.8 µg/kg), followed by goat's milk-based (52.7 ± 25.67), plant-based (48.8 ± 31.68), and cow's milk-based (27.5 ± 29.62) formulas (<i>p</i> < 0.001). The wide variability in AA concentrations among infant formulas can be attributed to differences in formulation, ingredient composition, manufacturing processes, and analytical methodologies. These findings highlight the need for continuous monitoring of AA levels in infant foods to ensure their safety.