Chromatographic Extraction and LC–Triple Quadrupole Mass Spectrometric Determination of Acrylamide in Powdered Infant Formulas

A recent joint study published in Molecules and conducted by researchers atAnkara Medipol University (Turkey) and the University of Almeria (Spain) presented a sensitive and selective analytical approach for acrylamide (AA) determination in powdered infant formulas (1). To reduce sample-handling steps and matrix interferences, an optimized solid–liquid extraction (SLE)-based extraction combined with a dispersive solid-phase extraction (d-SPE) clean-up and liquid chromatography–triple quadrupole mass spectrometry (LC-QqQ-MS) detection was implemented.

Commercially available infant formulas are commonly used as partial or complete substitutes for breast milk to feed infants and young children aged 0–36 months (2). There are two main forms of infant formula: ready-to-feed liquid and powdered (which must be reconstituted with water). Powdered is the most widely available, shelf-stable, and commercially distributed option of the two (3). Infant formulas are treated using a wide range of processing techniques, which use both thermal and non-thermal technologies, resulting in distinct final product compositions (3). Conventional thermal processes are necessary in the manufacturing of powdered products and for the realization of pasteurization or sterilization effects. Although these techniques successfully decrease microbial loads and extend the final product’s shelf life, maintaining a suitable balance between microbial safety and the prevention of unwanted chemical changes is critical (4,5).

During production of formula, the Maillard reaction may occur when protein- and sugar-containing ingredients are exposed to heat, resulting in the formation of harmful compounds such as AA, an α,β-unsaturated carbonyl compound formed in certain foods processed at temperatures above 120 °C as a result of the reaction between amino acids and the reduction ofsugars such as glucose and fructose (6,7). AA is classified as a Group 2A compound by the International Agency for Research on Cancer (IARC), meaning it is “probably carcinogenic to humans” (8).

“Since AA poses significant health hazards, including carcinogenicity and neurotoxicity, the application of effective mitigation and analytical strategies is essential to protect consumers and comply with international safety standards,” the authors wrote in their study (1). “AA represents a particularly high health risk for infants due to their greater daily dietary intake per kilogram of body weight, higher resting metabolic and respiratory rates, and larger body surface area compared with adults.”

The authors of the study reported that their 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) and plant-based (48.8 ± concentrations (1).

“These findings,” the authors write in their article (1), “highlight the need for continuous monitoring of AA levels 31.68), and cow's milk-based (27.5 ± 29.62) formulas (p < 0.001). The wide variability in AA in infant foods to ensure their safety.”

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References

1. Sevim, S.; Lopez-Ruiz, R.; Garrido-Frenich, A. Acrylamide Determination in Infant Formulas: A New Extraction Method. Molecules 2025, 30 (24), 4718. DOI: 10.3390/molecules30244718
2. WHO. Guidance on Ending the Inappropriate Promotion of Foods for Infants and Young Children: Implementation Manual. 2017. https://www.who.int/publications/i/item/9789241513470(accessed 2025-08-26).
3. Bakshi, S.; Paswan, V. K.; Yadav, S. P. et al. A Comprehensive Review on Infant Formula: Nutritional and Functional Constituents, Recent Trends in Processing and its Impact on Infants’ Gut Microbiota. Front. Nutr. 2023, 10, 1194679. DOI: 10.3389/fnut.2023.1194679
4. Jiang, S. L.; Guo, M. R. 8–Processing Technology for Infant Formula; In Human Milk Biochemistry and Infant Formula Manufacturing Technology; 2nd ed.; Guo, M., Ed. Woodhead Publishing, 2021; pp. 223–240. DOI: 10.1016/B978-0-08-102898-8.00008-8
5. Varghese, K. S.; Pandey, M. C.; Radhakrishna, K. et al. Technology, Applications and Modelling of Ohmic Heating: A Review. J. Food Sci. Technol. 2014, 51 (10), 2304-2317. DOI: 10.1007/s13197-012-0710-3
6. Sun, X.; Wang, C.; Wang, H. et al. Effects of Processing on Structure and Thermal Properties of Powdered Preterm Infant Formula. J. Food Sci. 2018, 83 (6), 1685-1694. DOI: 10.1111/1750-3841.14162
7. Aktağ, I. G.; Hamzalıoğlu, A.; Kocadağlı, T. et al. Dietary Exposure to Acrylamide: A Critical Appraisal on the Conversion of Disregarded Intermediates into Acrylamide and Possible Reactions During Digestion. CRFS 2022, 5, 1118–1126. DOI: 10.1016/j.crfs.2022.07.004
8. IARC. Acrylamide. In Monographs on the Evaluation of Carcinogenic Risks to Humans: Some Industrial Chemicals; International Agency for Research on Cancer–IARC: Lyon, France, 1994; 60, 389–433.