Innovative ultrasound-assisted and vacuum manifold-assisted pipette tip solid-phase extraction using MSU-2-C18 sorbent for the extraction and purification of glycoalkaloids in potato-based snacks prior to UHPLC-MS/MS analysis

A miniaturized and environmentally conscious analytical workflow was developed and validated for the extraction, purification, and quantification of major glycoalkaloids (α-solanine, α-chaconine), the degradation product γ-chaconine, and the aglycone solanidine in potato-based snack products. The method combines ultrasound-assisted extraction (UAE) with pipette-tip solid-phase extraction (PT-SPE), followed by UHPLC-MS/MS analysis. UAE conditions were optimized using a Box-Behnken statistical design, which evaluated the extraction time, ultrasound amplitude, and solid:liquid (SL) ratio. Optimal conditions were determined to be 5 min, 100% amplitude, and a 1:9 w/v of SL ratio, with methanol as the most effective extraction solvent. MSU-2-C18 mesostructured silica sorbent, packed into pipette tips (5 mg) connected to a SPE vacuum manifold (VMA-PT-SPE), was employed under the optimized conditions to simultaneously purify up to 12 extracts. The method's greenness was evaluated using the AGREEprep metric, yielding a score of 0.65, indicating good environmental performance. Limits of detection (LOD) and quantification (LOQ), ranging from 0.55-4.40 µg/kg and 1.82-14.67 µg/kg, respectively, were achieved for the target analytes. Application to four commercial potato-based snacks revealed major (α-solanine and α-chaconine) and minor glycoalkaloids (γ-chaconine and solanidine) levels, ranging from 2.0 to 16.0 mg/kg fresh weight, highlighting substantial variability among products and providing a more comprehensive characterization of the glycoalkaloid profile. Compared to conventional SPE formats, this UAE-VMA-PT-SPE strategy offers superior reproducibility while reducing sample and solvent consumption. The method represents a powerful tool for routine monitoring of glycoalkaloids in potato snacks, supporting food safety assessments and addressing analytical priorities established by the European Food Safety Authority.