Human Milk Fatty Acid Composition: Comparison of Novel Dried Milk Spot Versus Standard Liquid Extraction Methods

Abstract

Accurate assessment of the long chain polyunsaturated fatty acid (LC-PUFA) content of human milk (HM) provides a powerful means to evaluate the FA nutrient status of breastfed infants. The conventional standard for FA composition analysis of HM is liquid extraction, trans-methylation, and analyte detection resolved by gas chromatography. This standard approach requires fresh or frozen samples, storage in deep freeze, organic solvents, and specialized equipment in processing and analysis. Further, HM collection is often impractical for many studies in the free living environment, particularly for studies in developing countries. In the present study, we compare a novel and more practical approach to sample collection and processing that involves the spotting and drying ∼50 μL of HM on a specialized paper stored and transported at ambient temperatures until analysis. Deming regression indicated the two methods aligned very well for all LC-PUFA and the abundant HM FA. Additionally, strong correlations (r>0.85) were observed for DHA, ARA, EPA, linoleic (LA), and alpha-linolenic acids (ALA), which are of particular interest to the health of the developing infant. Taken together, our data suggest this more practical and inexpensive method of collection, storage, and transport of HM milk samples could dramatically facilitate studies of HM, as well as understanding its lipid composition influences on human health and development.

The dried blood spot sampling method in the laboratory medicine

“…Self-sampling, simple collection and ease of transportation are big pluses of this method. Dried milk spot (DMS) samples can be used also for TDM (109)(110)(111).…”

Low Neonatal Plasma n-6/n-3 PUFA Ratios Regulate Offspring Adipogenic Potential and Condition Adult Obesity Resistance

“…Total lipids were extracted and fatty acid profiles quantified by gas chromatography-mass spectrometry as previously described (14,25). Data are expressed in micromoles of fatty acid per milliliter of milk or milligram of total plasma protein.…”

Comparison of breast milk fatty acid composition from mothers of premature infants of three countries using novel dried milk spot technology

“…The DMS method described in this study allows the measurements of breast milk fatty acid composition more accurately with even less sample required than in the studies published previously [8,9]. The current DMS has advantages compared to the methodologies previously published, as it consists of three protecting components that have been proven to be necessary to stabilise LCPUFA [10].…”

“…The current DMS has advantages compared to the methodologies previously published, as it consists of three protecting components that have been proven to be necessary to stabilise LCPUFA [10]. The filter paper used in this study was silica gel impregnated, which allows stronger binding with the lipids than the non-silica coated Ahlstrom (Grade 226) filter paper used in previous studies [8,9], and the addition of chelating agent that used in our PUFAcoat TM system offers more protection from oxidation.…”

“…One such technique has been developed based on the current dried blood technology [6,7], namely the dried milk spot (DMS) technology, which has been commercialized [8,9]. The method only requires a single drop of breast milk, simplifies the analytical process, and does not require cold-chain transportation and storage.…”