Triacylglycerol structure determination

Solvent-free enzymatic interesterification of milk fat alone or with other fats or fatty acids provides the most acceptable route for modification of the triacylglycerol structures in milk fat and further research and development in this field is expected to provide physical and physiological benefits. From a nutritional perspective, it is of interest to examine the effects of randomized milk fat on serum cholesterol. Christophe et al. (1978) reported that substitution of native milk fat with chemically-randomized interester-ified milk fat reduced cholesterol levels in man. However, others found that there was no effect on serum cholesterol levels in man as a result of substitution of ezymatically randomized milk fat (De Greyt and Huyghebaert, 1995). Further studies are required to determine if interesterilied milk fat provides a nutritional benefit. 

General Analysis of trans fatty acids NMR spectra of lipids Guidelines for interpretation of 13C NMR spectra Applications of high-resolution methods to oils and fats Structure determination of polyunsaturated triacylglycerols 31P NMR for profiling of phospholipids 1998 (210) 1996(211) 1998(10, 18) 1999(212,213) 1999(213) 1998 (10) 1996(214) 1996(215)... 

Santinelli, F., Damiani, R and Christie, W.W. (1992) The triacylglycerol structure of olive oil determined by silver ion high performance liquid chromatography in combination with stereospecific analysis. J. Am. Oil Chem. Soc., 69, 552-557. 

Alternatively, triacylglycerol structure can be determined by chemical reaction based on a Grignard reagent (e.g. ethyl magnesium bromide), which yields j -l(3),2-diacylglycerols. This method is more reliable, but is tedious and time-consuming (Brockerhoff, 1967). 

Pham, L.J., Gregorio, M.A., 2008. The triacylglycerol structure of coconut oil determined by chromatography combined with staeospecific analysis. Philipp. Agric. Scd. 91 (3), 343-347. 

Using PTLC six major fractions of lipids (phospholipids, free sterols, free fatty acids, triacylglycerols, methyl esters, and sterol esters) were separated from the skin lipids of chicken to smdy the penetration responses of Schistosoma cercaria and Austrobilharzia variglandis [79a]. To determine the structure of nontoxic lipids in lipopolysaccharides of Salmonella typhimurium, monophosphoryl lipids were separated from these lipids using PTLC. The separated fractions were used in FAB-MS to determine [3-hydroxymyristic acid, lauric acid, and 3-hydroxymyristic acids [79b]. 

Certain classes of lipids are susceptible to degradation under specific conditions. For example, all ester-linked fatty acids in triacylglycerols, phospholipids, and sterol esters are released by mild acid or alkaline treatment, and somewhat harsher hydrolysis conditions release amide-bound fatty acids from sphingolipids. Enzymes that specifically hydrolyze certain lipids are also useful in the determination of lipid structure. Phospholipases A, C, and D (Fig. 10-15) each split particular bonds in phospholipids and yield products with characteristic solubilities and chromatographic behaviors. Phospholipase C, for example, releases a water-soluble phosphoryl alcohol (such as phosphocholine from phosphatidylcholine) and a chloroform-soluble diacylglycerol, each of which can be characterized separately to determine the structure of the intact phospholipid. The combination of specific hydrolysis with characterization of the products by thin-layer, gas-liquid, or high-performance liquid chromatography often allows determination of a lipid structure. 

The position of fatty acids in the triacylglycerol (TAG) structure not only significantly influences fatty acid absorption, but could provide important markers for establishing the authenticity of different oils. Lawson and Hughes (1988) determined the TAG stereospecific structure of a number of GLA rich oils,... 

The nature of the fatty acids in TAGs determines their hydrophobicity/hydrophih-city and diffusional mobility. In an aqueous/hpid environment, such as adipose tissue or lipoproteins in plasma, the relative hydrophihcity of the TAGs determines their partitioning between the interfacial phase and the apolar phase. This may have far stretching consequences. For instance, the rate and selectivity of fatty acid mobilization from fat cells may affect levels and composition of the nonester-ified fatty acids in plasma. These in mrn affect lipid homeostasis. Rate and selectivity of fatty acid mobilization from adipose stores are not related to the positional distribution of fatty acids on the glycerol backbone (75). They are related, however, to triacylglycerol hydrophihcity and thus to TAG structure (76). 

Cholesterol, which is largely insoluble in aqueous m a, travels through the blood circulation in the form of Upoprotein complexes. The plasma lipoproteins are a family of globular particles that share common structural features. A core of hydrophobic lipid, principally triacylglycerols (triglycerides) and cholesterol esters, is surrounded by a hydrophilic monolayer of phospholipid and protein (the apolipoproteins) [1-3]. Lipid-apolipoprotein interactions, facihtated byi amphi-pathic protein helices that segregate polar from nonpolar surfaces [2,3], provide the mechanism by which cholesterol can circulate in a soluble form. In addition, the apolipoproteins modulate the activities of certain enzymes involved in Upoprotein metabolism and interact with specific cell surface receptors which take up Upopro-teins by receptor-mediated endocytosis. Differences in the Upid and apoUpoprotein compositions of plasma Upoproteins determine their target sites and classification based on buoyant density. 

There are a number of books available that deal with lipids and their structures, and the author has found those cited to be of particular value [319,367]. Literally thousands of papers have appeared over the last 25 years detailing the structures and compositions of lipids from particular tissues and species, as determined by modem chromatographic methods, but there appears to have been very little effort to collate and critically compare these data in any systematic way, or to relate the compositions of lipids to their functions. Among other consequences of this, there remain anomalies and gaps in our knowledge. Comprehensive accounts of the lipids of the tissues of ruminant animals [162], tissue and membrane phospholipid compositions [395,970] and triacylglycerol compositions [125,553,686,824] have appeared, however, and there are miscellaneous reviews of the compositions of specific lipid classes or tissues in the literature. The author recently attempted to summarise the essential features of lipid composition in a succinct manner [168]. This cannot be repeated here, and a brief summary only of lipid structure and composition follows. 

EI-MS with direct probe insertion was used, for example, to determine the structure of unusual tetraacylglycerols containing an allenic acid [877] and triacylglycerols containing sorbic acid [251]. Molecular species of mixed triacylglycerols have also... 

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