This study presents a novel environmentally-friendly process for the selective extraction and enrichment of DHA/EPA-containing phospholipids (PL-DHA/EPA) from krill oil. The methodology leverages differential crystallization behavior between phospholipids and triacylglycerols in ethanolic solutions, exploiting their distinct freezing point thresholds to achieve precise fractionation. Response surface methodology optimization identified optimal extraction parameters: liquid-to-material ratio of 6:1 (v/w), freezing temperature of −20 °C, freezing duration of 25 h, and rotary evaporation temperature of 45 °C, yielding a final product with 39.40% PL-DHA/EPA content. Principal component analysis revealed substantial overlap in confidence ellipses among extraction methodologies, indicating effective preservation of core phospholipid signatures from the parent krill oil while maintaining critical structural characteristics and molecular species distribution. Comprehensive analysis of phospholipid fractions and heatmap analysis revealed distinctive molecular profiles compared to conventional organic solvent extraction, with selective enrichment of EPA-containing phospholipids, particularly PC-EPA and PI-EPA species. The green extraction method demonstrated comparable oxidative stability to conventional approaches, with superior protection against secondary oxidation as evidenced by significantly lower anisidine values. This sustainable approach achieves effective phospholipid enrichment while substantially reducing environmental impact through elimination of halogenated solvents, addressing the critical need for environmentally conscious technologies in marine lipid processing with potential applications in nutraceutical and functional food industries.

Antarctic krill oil (AKO) is a valuable nutraceutical; however, it is highly susceptible to oxidation. Encapsulation represents an effective strategy to enhance the storage stability of AKO. This study explored a novel approach for encapsulating AKO using sodium alginate (ALG) and gelatin (GLN) to improve its stability, and multiple parameters were systematically evaluated, including oil-loading efficiency, surface oil content, particle size, water activity, and thermal stability. Additionally, core-material retention efficiency, acid value, peroxide value, and anisidine value were measured after accelerated oxidation. The results demonstrated that the optimal encapsulation conditions consisted of an ALG:GLN ratio of 2:1, a 9% CaCl₂ coagulation bath, 750 μm nozzle size, followed by freeze-drying. Under these conditions, the microcapsules achieved an oil-loading efficiency of 62.63% and a surface oil content of 19.21%. The water activity of the microcapsules was 0.516. Thermogravimetric analysis indicated that AKO microcapsules encapsulated with ALG/GLN exhibited higher thermal stability (~300 °C) compared to those encapsulated with ALG alone (~280 °C). When AKO or its microcapsules were subjected to accelerated oxidation at 65 °C, compared to ALG-encapsulation alone, the ALG/GLN encapsulation system significantly reduced the oxidation indicators of the oil, such as acid value (24%), peroxide value (26%), and anisidine value (28%). In conclusion, incorporating GLN into ALG-based microcapsules significantly enhanced the antioxidant capacity of AKO and prolonged its shelf life.