Selective extraction of phospholipids from whey protein phospholipid concentrate using supercritical carbon dioxide...

In recent years, using dairy phospholipids (PL) as functional ingredients has increased because PL have nutritional benefits and functional properties. In this study, a novel 2-step supercritical fluid extraction (SFE) process was used to extract whey protein phospholipid concentrate (WPPC), a dairy co-product obtained during the manufacture of whey protein isolate, for PL enrichment. In the first step, nonpolar lipids in WPPC were removed using neat supercritical carbon dioxide (S-CO₂) at 41.4 MPa and 60°C. In the second stage, the feasibility of using the polar solvent ethanol as a co-solvent to increase the solubility of S-CO₂ extraction solvent was explored. A 3 × 3 × 2 factorial design with extraction pressure (35.0, 41.4, and 55.0 MPa), temperature (40 and 60°C), and concentration of ethanol (10, 15, and 20%) as independent factors was used to evaluate the extraction efficiency providing the most total PL, and the best proportion of each individual PL from the spent solids collected during S-CO₂ SFE. All lipid fractions were analyzed using thin-layer chromatography and high-performance lipid chromatography. The total amount of PL extracted from WPPC was significantly affected by ethanol concentration; the extraction pressure and temperature were nonsignificant. The optimal SFE condition for generating a concentrated PL lipid fraction was 35.0 MPa, 40°C, and 15% ethanol concentration; the highest amount of extracted PL averaged 26.26 g/100 g of fat. Moreover, adjusting SFE condition allowed successful recovery of a high concentration of sphingomyelin, phosphatidylcholine, and phosphatidylethanolamine, giving averages of 11.07, 10.07, and 7.2 g/100 g of fat, respectively, 2 to 3 times more than conventional solvent extraction. In addition, exhausted solids obtained after the SFE process were enriched with denatured proteins (72% on dry basis) with significantly more water-holding capacity and emulsifying capacity than untreated WPPC. Overall, this 2-stage SFE process using neat S-CO₂ and ethanol has the greatest potential to produce a PL-rich lipid fraction from WPPC.