A Novel ATPS-HPLC(GC) method for the rapid Quantification of dehydroacetic acid in food

Dehydroacetic acid (DHA) is a commonly used preservative widely applied in food products. Currently, the quantitative analysis process for DHA remains cumbersome, involving multiple toxic organic reagents. To address these issues, this study leveraged the low-toxicity, environmentally friendly, and rapid characteristics of aqueous two-phase systems (ATPS) to develop a pretreatment method suitable for the extraction and purification of DHA. This method is compatible with both high-performance liquid chromatography (HPLC) and gas chromatography (GC) techniques. For HPLC, acidic chromatographic conditions were developed to mitigate issues such as peak tailing and retention time drift, improving peak shapes and stabilizing retention times. For GC, appropriate chromatographic conditions were established for accurate quantitative analysis of DHA. The study focused on screening appropriate ATPS types and optimizing the composition ratios of their constituent components to achieve optimal extraction and purification efficiency for DHA. Method validation and sample analysis were conducted for both ATPS-HPLC and ATPS-GC approaches. Under the ATPS conditions composed of 5.00 mL water, 2.00 mL n-propanol, and 7.00 g sodium dihydrogen phosphate, results demonstrated that the ATPS-HPLC (GC) method is accurate, reliable, rapid, and environmentally friendly, with detection limits of 2.00 × 10⁻⁴ g/kg (6.00 × 10⁻⁴ g/kg), average recoveries of 93.7–104 % (98.3–107 %), and relative standard deviations (RSDs) of 0.7–5.2 % (1.9–6.8 %).