Preparation of human milk fat analogue by enzymatic interesterification reaction using palm stearin and fish oil.

Palm stearin fractionate (PSF), obtained from palm stearin by further fractionation with solvents and n-3 polyunsaturated fatty acids (n-3 PUFA) rich fish oil (FO) were subjected to interesterification at 1:1, 1:2, 1:3, 2:1 and 3:1 substrate molar ratio and catalyzed by lipase from Thermomyces lanuginosa for obtaining a product with triacylglycerol (TAG) structure similar to that of human milk fat (HMF). The parameters (molar ratio and time) of the interesterification reaction were standardized. The temperature of 60 °C and enzyme concentration of 10 % (w/w) were kept fixed as these parameters were previously optimized. The reactions were carried out in a stirred tank reactor equipped with a magnetic stirrer for 6, 12, 18 and 24 h. The blends were analyzed for fatty acid (FA) composition of both total FAs and those at the sn-2 position after pancreatic lipase hydrolysis. All the blended products were subjected to melting point determination and free fatty acid content. Finally, blend of PSF and FO at 2:1 molar ratio with 69.70 % palmitic acid (PA) content and 12 h of reaction produced the desired product with 75.98 % of PA at sn-2 position, 0.27 % arachidonic acid (AA), 3.43 % eicosapentaenoic acid (EPA) and 4.25 % docosahexaenoic acid (DHA) and with melting point of 42 °C. This study portrayed a successful preparation of TAG containing unique FA composition i.e. ≥ 70 % of the PA, by weight, were esterified at the sn-2 position which could be used in infant formulation with health benefits of n-3 PUFAs.