Evaluation of Extraction Techniques for Recovery of Microalgal Lipids under Different Growth Conditions.

Microalgal lipids contain a wide array of liposoluble bioactive compounds, but lipid extraction remains a critical limitation for their commercial use. An accelerated solvent extraction (ASE) was used to extract lipids from Chlamydomonas reinhardtii, Arthrospira platensis (Spirulina), and Chlorella vulgaris grown under either standard or nitrogen depletion conditions. Under standard growth conditions, ASE using methanol:chloroform (2:1), methyl tert-butyl ether (MTBE):methanol:water, and ethanol at 100 °C resulted in the highest recovery of total lipids (352 ± 30, 410 ± 32, and 127 ± 15 mg/g biomass from C. reinhardtii, C. vulgaris, and A. platensis, respectively). Similarly, the highest total lipid and triacylglycerols (TAGs) recovery from biomass cultivated under nitrogen depletion conditions was found at 100 °C using methanol:chloroform, for C. reinhardtii (total, 550 ± 21; TAG, 205 ± 2 mg/g biomass) and for C. vulgaris (total, 612 ± 29 mg/g; TAG, 253 ± 7 mg/g biomass). ASE with MTBE:methanol:water at 100 °C yielded similar TAG recovery for C. reinhardtii (159 ± 6 mg/g) and C. vulgaris (200 ± 4 mg/g). Thus, MTBE:methanol:water is suggested as an alternative substitute to replace hazardous solvent mixtures for TAGs extraction with a much lower environmental impact. The extracted microalgal TAGs were rich in palmitic (C16:0), stearic (C18:0), oleic (C18:1,9), linoleic (C18:2n6), and α-linolenic (C18:3n3) acids. Under nitrogen depletion conditions, increased palmitic acid (C16:0) recovery up to 2-fold was recorded from the biomasses of C. reinhardtii and C. vulgaris. This study demonstrates a clear linkage between the extraction conditions applied and total lipid and TAG recovery.

Structural characteristics of triacylglycerols contribute to the distinct in vitro gastric digestibility of sheep and cow milk fat prior to and after homogenisation.

The regiodistribution of fatty acids (FAs) in triacylglycerols (TAGs) is highly correlated to the digestion and absorption of dietary fats. During in vitro gastric digestion, medium-long-medium/short chain triacylglycerols (MLM) showed the fastest digestibility among 105 TAG molecular species, for both raw and homogenised milk, resulting from the higher activity of gastric lipase towards medium-chain fatty acids (MCFAs) than towards long-chain fatty acids (LCFAs). After 60 min of in vitro gastric digestion, the percentage of digestion of each MLM was 16.6-31.2% and 12.3-18.6% respectively in raw sheep and cow milk, and it reached 33.4-45.5% and 23.0-38.9% respectively, after homogenisation. Sheep milk TAGs were digested faster than cow milk TAGs in raw milk (16.1% and 12.4%, respectively) as well as after homogenisation (24.1% and 20.3%, respectively). This is due to the higher percentages of MCFAs esterified on the sn-1/3 positions of sheep milk TAGs (48.3-67.3% and 11.9-51.5% respectively for sheep and cow milk prior to and after homogenisation). Homogenisation favoured TAG lipolysis due to the increased fat globule surface area; however, it did not affect lipase regiospecificity towards TAG molecular species.

In-depth lipidomic analysis of tri-, di-, and mono-acylglycerols released from milk fat after in vitro digestion.

Milk fat is arguably one of the most complex fats found in nature and varies widely between animal species. Analysis of its digestion products is tremendously challenging, due to the complexity, diversity, and large range of concentrations of triacylglycerols (TAGs) and their digestion products (i.e. diacylglycerols (DAGs), monoacylglycerols (MAGs), and free fatty acids (FFAs)). Therefore, a method combined the solid phase extraction (SPE), high-performance liquid chromatography (HPLC) and multi-dimension mass spectrometry (MDMS) was developed to identify and semi-quantify the TAGs, DAGs and MAGs in milk fat after in vitro digestion. Up to 105, 64, 14 and 30 species of TAGs, DAGs, MAGs, and FFAs were determined with their concentrations of 0.01-22.3, 0.01-39.2, 0.01-47.8, and 0.04-191.0 mg/g fat, respectively, during the in vitro digestion of cow and sheep milk. The validation of the method shows that this method was precise and reliable.

Effects of season and industrial processes on volatile 4-alkyl-branched chain fatty acids in sheep milk.

4-alkyl-branched-chain fatty acids (vBCFAs) are responsible for the goaty-sheepy flavour of sheep and goat milk. The levels of vBCFAs were investigated in sheep, goat and cow milk powders. The effect of season and industrial processes on the levels of vBCFAs in sheep milk was also evaluated. Sheep milk powders contained lower amounts of 4-Me-8:0 and 4-Et-8:0 (143-176 and 1.2-10.7 µg/g milk fat) than goat milk powders (335-481 and 73.1-105 µg/g milk fat), while 4-Me-9:0 was low in both sheep and goat milk powders (<8 µg/g milk fat). Odour activity values (OAVs) of sheep milk powders (17-393) were 6-fold lower than the values of goat milk powders (882-1323). Higher levels of vBCFAs were found in sheep milk produced in spring than in summer. Thermisation had little impact on vBCFAs; however, spray drying led to dramatic increases in free vBCFAs, particularly towards late lactation, which resulted in higher OAVs in sheep milk powder compared to raw milk.

Effect of animal age on the nutritional and physicochemical qualities of ovine bresaola.

Bresaola made from New Zealand mutton and lamb, were compared in quality attributes. Mutton bresaola had slightly lower protein and higher moisture contents, and similar amount of intramuscular fat and instrumental colour compared to that of the lamb. 36 and 80 unique endogenous peptides were observed in mutton and lamb, respectively. Proteolysis during dry-curing was affected by the age of the animal and has resulted in softer and easier to chew textural properties to the mutton bresaola. Significantly higher amounts of total free amino acids and higher proportion of essential amino acids were detected in the mutton bresaola from digestion simulation compared to that from lamb. Expedited proteolysis measured in the form of the release of free amino acids was positively related to the animal age. Overall, bresaola from mutton had more favourable characteristics compared to that of the lamb.

Distribution of fatty acids and phospholipids in different table cuts and co-products from New Zealand pasture-fed Wagyu-dairy cross beef cattle.

Wagyu beef products are marketed as luxury goods to discerning consumers and the lipid content and composition are important drivers of wagyu product value. Wagyu beef is an extensively marbled meat product, well characterised for its tenderness and flavour. In New Zealand, pasture-fed Wagyu-dairy beef production is increasing to meet demand for ultra-premium meat products. Important for these characteristics is the composition of lipid species and their distribution across the carcass. The aim of this study was to analyse the distribution of fatty acids and phospholipids in 26 table cuts, nine co-products and three fat deposits of carcasses from New Zealand pasture-fed Wagyu-dairy cross beef carcasses (n = 5). Phospholipid and fatty acid levels varied across different cuts of the carcass, but typically cuts with high levels of phospholipids also had high levels of omega-3 fatty acids and low levels of saturated fatty acids. This work will be used in the future to examine the potential health aspects of pasture-fed Wagyu beef.

Isotopic labeling of milk disialogangliosides (GD3).

The most abundant ganglioside group in both human milk and bovine milk during the first postnatal week is ganglioside GD3. This group of disialogangliosides forms up to 80% of the total ganglioside content of colostrum. Although dietary gangliosides have shown biological activity such as improvement of cognitive development, gastrointestinal health, and immune function, there is still a gap in our understanding of the molecular mechanisms governing its uptake and the metabolic processes affecting its bioavailability. The use of isotopically labeled ganglioside to track the bioavailability, absorption, distribution, and metabolism of gangliosides may provide key information to bridge this gap. However, isotope labeled GD3 is not commercially available and its preparation has not been described. We report for the first time the preparation of labeled GD3 with stable isotopes. Using alkaline hydrolysis, we were able to selectively remove both acetyl groups from the tetrasaccharide portion of GD3 without promoting significant hydrolysis of the ceramide portion of the molecule to generate N-deacetyl-GD3 (Neu5α2-8Neu5-GD3). The N-deacetyl-GD3 was then chemoselectively re-acetylated in aqueous medium using deuterated acetic anhydride in the presence of Triton X 100 to produce 2H6-GD3 {GD3[(Neu5Ac-11-2H3)-(Neu5Ac-11-2H3)]}. This method provided 2H6-GD3 with approximately 60% yield. This compound was characterized by proton nuclear magnetic resonance (1H NMR) and liquid chromatography mass spectrometry (LC-MS). The oral absorption of the 2H6-GD3 was demonstrated using a Sprague-Dawley weaning rats. Our results indicate that some ingested labeled milk gangliosides are absorbed and transported into the bloodstream without modification.