Simultaneous determination of acrylamide, Glycidamide, and Methylacrylamide in plant-based beverages using μ-dispersive solid phase extraction and HPLC-PDA: Development, validation, and occurrence

Plant-based beverages are widely consumed as dairy alternatives due to their nutritional value and sustainability. However, thermal processing may lead to the formation of acrylamide and related toxicants. This study developed and validated a miniaturized dispersive solid-phase extraction (μ-dSPE) method coupled with highperformance liquid chromatography and ultraviolet detection for the simultaneous determination of acrylamide, glycidamide, and methacrylamide, using thiourea as an internal standard. Unlike existing μ-dSPE/ QuEChERS and MS-based approaches, the workflow is entirely aqueous (no organic solvents) and requires no derivatization, aligning with green analytical chemistry. Optimization was conducted using a 34 41 screening design and response surface methodology. The method showed excellent linearity (r2 > 0.999), low detection limits (0.24–0.98 μg/mL), and excellent accuracy (91–111 % recovery) and precision (RSD < 6.1 %). Analysis of 15 commercial beverages revealed acrylamide in 14 samples (0.33–1.17 μg/mL), especially in oat and mixedprotein types; glycidamide and methacrylamide were not detected. Environmental assessment using AGREEprep yielded a high greenness score (0.72) and a high practical applicability score (80.0) by the Blue Applicability Grade Index. This μ-dSPE-HPLC-PDA method provides a reliable, cost-effective, and environmentallyfriendly approach for monitoring heat-induced contaminants in plant-based beverages.