Abstract
Human milk (HM) is extremely important to the baby, containing a varied and balanced range of nutrients fundamental to the ideal development of the newborn. Among these nutrients, HM lipids provide 40-60% of the energy. In HM banks, the creamatocrit method is employed as a quick and solvent-free tool, however, since Folch methodology is considered reference for lipid extraction, this study compared the fatty acid (FA) composition and the lipid profile by gas chromatography with flame ionization detector (GC-FID) and direct infusion by electrospray ionization mass spectrometry (DI-ESI-MS) in pools of different HM phases, obtained from Folch and creamatocrit methods, in order to elucidate which method provides better information about HM lipids. Lipid compositon results by Folch were superior to the results obtained by creamatocrit in HM pools investigated. It means, the lipid profiles of all HM pools were more intense, and the number of FA identified by Folch was also higher in comparison to the creamatocrit method (21 and 6 FAs, respectively). GC-FID analysis in combination with principal component analysis (PCA) divided the methods, confirming a greater contribution of the Folch method. Therefore, Folch methodology revealed an effective lipid extraction in comparison to creamatocrit method.
Highlights
Human milk (HM) is considered the gold standard food to assemble the baby requirements due to the varied and balanced range of nutrients, as well as several components dispersed in the colloid liquid, which are fundamental to the ideal development of the newborn.[1,2] Until the 7th day after delivery the HM is considered colostrum, between the 8th and 14th day postpartum it is considered transitional milk and after the 15th day postpartum, it is considered mature milk.[3]HM lipids are extremely important for the newborn proper growth, providing 40-60% of the required energy
The creamatocrit lipid extraction method in the different phases of HM pools (colostrum (CrC); transitional (CrT); mature (CrM)) contributed positively to PC1 and led to the formation of a fifth group, separated from all other fatty acid (FA), since the results on the FA composition by this method were lower in comparison to those obtained by the Folch method
Creamatocrit method is a lipid extraction methodology applied in human milk banks even though Folch method is considered reference for lipid extraction
Summary
Human milk (HM) is considered the gold standard food to assemble the baby requirements due to the varied and balanced range of nutrients, as well as several components dispersed in the colloid liquid, which are fundamental to the ideal development of the newborn.[1,2] Until the 7th day after delivery the HM is considered colostrum, between the 8th and 14th day postpartum it is considered transitional milk and after the 15th day postpartum, it is considered mature milk.[3]. 1064 Lipid Profile Determination by Direct Infusion ESI-MS and Fatty Acid Composition by GC-FID in Human Milk Pools J. Folch methodology is considered reference for lipid extraction, so this study compared the fat content and the FA composition by gas chromatography with flame ionization detection (GC-FID), and the lipid profiles by direct infusion by electrospray ionization mass spectrometry (DI-ESI-MS) in colostrum, transitional and mature HM pool samples, obtained from Folch and creamatocrit methods, in order to elucidate which method provides better information about the HM lipids. Folch method For the analysis by Folch method, 10 mL of HM samples from each pool (colostrum, transitional and mature) in triplicate were used to obtain 125 mg of lipids for esterification/transesterification and subsequent identification of fatty acids methyl esters (FAMEs) by GC-FID and analysis of the lipid profile by DI-ESI-MS. Results obtained by GC-FID were performed to principal component analysis (PCA) using RStudio software.[20]
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