| Summary: | 2’-Fucosyllactose (2’-FL) is the predominant oligosaccharide in human milk, responsible for important infant biological functions. Its quantification requires specific extraction and analysis methods, but current techniques use toxic solvents for extraction and are time-consuming. Therefore, this study optimized the extraction of 2’-FL from mature human milk, using water to eliminate the toxic solvents. Using response surface methodology, 27 experiments were evaluated by high-performance liquid chromatography coupled to mass spectrometry, with their concentrations calculated based on a standard curve. The optimal conditions included initial centrifugation time (15 min), water volume (3.0 mL), incubation time (90 min), and final centrifugation time (40 min), resulting in a maximum concentration of 3046 mg L–1. The method was validated in the other lactation phases, representing 1892.93 mg L–1 (transition) to 4247.33 mg L–1 (colostrum), showing high reproducibility and agreement with the literature. The proposed method was 50.43% faster and more economical than traditional methods, standing out as an efficient approach for the 2’-FL extraction.
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