A novel non-derivatization HPLC/UV method for the determination of some n-3 free fatty acids in breast milk matrix

Abstract

The mother's milk is a valuable source of many nutrients, including fatty acids (FAs). The most important fatty acids are polyunsaturated fatty acids (PUFAs), including linoleic and alpha-linolenic acids (ALA) that are essential for humans, as well as fatty acids that are formed by desaturation and elongation, such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) that occur in small amounts in breast milk. The content of exogenous fatty acids in milk is influenced by maternal age, genetics, hormones, infections, metabolic disorders, medications, dietary habits, supplementation, adiposity, lactation stage, feeding method, intervals between breastfeeding, and duration of breastfeeding.The proposed method for isolating fatty acids from breast milk has a recovery rate of 80–92% (SD of 4–9). Most importantly, derivatization to methyl esters is not necessary. Due to low limits of detection (e.g. 0.009 µg/mL of milk for DHA) and quantification (e.g. 0.090 µg/mL of milk for DHA), as well as high linearity (R2 = 0.9997), this method can be effectively used to identify and quantify three unsaturated fatty acids in breast milk using HPLC/UV. The developed method for the isolation and determination of PUFAs in breast milk can be widely applied in routine research, e.g. in milk banks.