Nonpolar Triglycerides Research Articles

Imaging of triglycerides in tissues using nanospray desorption electrospray ionization (Nano-DESI) mass spectrometry

Nonpolar triglycerides (TGs) are rarely detected in mass spectrometry imaging (MSI) experiments unless they are abundant in the sample. Herein, we use nanospray desorption electrospray ionization (nano-DESI) to explore the role of the solvent composition and ionic dopants on the detection of TGs in a murine gastrocnemius muscle tissue used as a model system. We evaluated three solvent mixtures for their ability to extract nonpolar TG species: MeOH:H2O 9:1 (v/v), MeOH:DCM 6:4 (v/v) and MeOH:AcN:tol 5:3.5:1.5 (v/v/v). We observe that TGs are mainly detected as [M+K]+ adducts and their extraction efficiency is improved using less polar solvents: MeOH:DCM and MeOH:AcN:tol. We also explore whether the ionization efficiency of TGs may be improved by doping the MeOH:AcN:tol solvent with ammonium formate (AF) and other ionic additives. However, the formation of [M+NH4]+ adducts of TGs is less efficient than that of [M+K]+ adducts in the range of AF concentrations from 0.1 to 10 mM. Chemical derivatization using 100 μM of Girard T reagent predominately generates reaction products of phosphatidylcholines rather than TG species. Moreover, the presence of the Girard T reagent suppresses ion signals of all the species in the spectrum including TGs. Nano-DESI MSI experiments performed using MeOH:AcN:tol solvent enable imaging of TGs without any detectable adverse effect on signals of other lipids and metabolites. Specifically, 10 out of 14 TG species were detected exclusively using MeOH:AcN:tol and the sensitivity towards other TGs was improved by at least an order of magnitude. Although polyunsaturated TGs can be detected using both solvents, saturated and monounsaturated TGs are only detected using MeOH:AcN:tol. Our results provide a direct path for the improved detection of TGs in tissue imaging experiments using liquid-based ambient ionization techniques.

Три варианта переноса жирных кислот в форме триглицеридов у плотоядных, травоядных до синтеза инсулина и при действии гормона инсулина

Является ли человек всеядным? Биологически это невозможно; Homo sapiens - травояден. В филогенезе же предки человека при жизни в океане, были плотоядными, рыбоядными. Предполагается, что в филогенезе последовательно с интервалами в миллионы лет сформировались 3 системы переноса к клеткам жирных кислот в форме неполярных триглицеридов в составе апоВ липопротеинов. 1. У плотоядных (Carnivore) в океане и на суше: энтероциты, апоЕ/В-48 хиломикроны → гепатоциты→ апоВ-100 ЛПОНП→ ЛПНП→ апоВ-100 эндоцитоз. 2. У травоядных (Herbivore), до синтеза инсулина перенос химически инертной пальмитиновой насыщенных жирных кислот осуществлялся по пути: гепатоциты→ пальмитиновые ЛПОНП→ ЛПНП→ апоВ-100 поглощение клетками. 3. У травоядных (Herbivore) при действии инсулина перенос химически активной олеиновой мононенасыщенной жирной кислоты представлял наиболее короткий путь: гепатоциты→ олеиновые ЛПОНП→ апоЕ/В-100 эндоцитоз инсулинзависимыми клетками. На ступенях филогенеза; 1. у плотоядных, 2. у травоядных до синтеза инсулина и 3. у травоядных при действии инсулина: 1. хиломикроны → пальмитиновые ЛПОНП→ ЛПНП→ апоВ-100 эндоцитоз; 2. пальмитиновые ЛПОНП→ ЛПНП→апоВ-100 эндоцитоз и 3. олеиновые ЛПОНП→ апоЕ/В-100 эндоцитоз, видно, почему олеиновые ЛПОНП не могут переносить пальмитиновые ЛПОНП с иными физико-химическими свойствами. В инсулинзависимой системе переноса олеиновой мононенасыщенной жирной кислоты, триглицериды задействованы только ЛПОНП и нет ЛПНП. При избытке плотоядной (мясной) пищи и пальмитиновой насыщенной жирной кислоты НЖК клетки не поглощают безлигандные пальмитиновые ЛПОНП→ЛПНП путем апоЕ/В-100 эндоцитоза. Формирование у пациента переноса насыщенных жирных кислот до действия инсулина объективизируется по уровню гиперлипопротеинемии (ГЛП) типа IIб. При избыточном потреблении мясной пищи и формировании переноса насыщенных жирных кислот, что характерно для плотоядных, у пациентов методом электрофореза выявляется гиперлипопротеинемия типа V. Атеросклероз и атероматоз, хотя и имеют общие звенья патогенеза, являются биологически разными, последовательными процессами, отражающими нарушение биологической функции трофологии и функции эндоэкологии. Is the human being an omnivore? Biologically, this is impossible. Homo sapiens is the herbivore. However phylogenetically, ocean-living ancestors of people were fish-eating carnivores. We suggest that three systems of fatty acid (FA) transport to cells have formed successively, at several million-year intervals, in the form of nonpolar triglycerides (TG) as constituents of apoB lipoproteins (LP): i) in the carnivores, both in the ocean and on land: enterocytes → apoВ-100 VLDL → LDL → apoВ-100 endocytosis; ii) in herbivores prior to insulin production: transport of chemically inert palmitic FA along the hepatocytes → palmitic VLDL→ LDL→ apoВ-100 cellular uptake pathway; iii) in herbivores under the action of insulin, the transport pathway for chemically active oleic FA is the shortest: hepatocytes→ oleic VLDL→ apoЕ/В-100 endocytosis by insulin-dependent cells. Thus, oleic VLDL cannot transport palmitic TG that have different physico-chemical parameters. In the insulin-dependent transport of oleic FA, TG are associated only with VLDL but not with LDL. When the diet is rich in meat and palmitic saturated FA cells do not internalize nonligand palmitic VLDL→LDL via apoЕ/В-100 endocytosis; physiologically, this endocytosis does not exist. In type IIb hyperlipoproteinemia, saturated FA are transported before the effect of insulin. If a patient eats excessive meat, transport of saturated FA follows the pathway typical of carnivore animals, which results in type V hyperproteinemia as detected by LP electrophoresis. Although atherosclerosis and atheromatosis share some pathogenetic features they are biologically different processes associated with impaired biological functions of trophology and endoecology.

The becoming of reactions of lipolysis in phylogenesis. The palmitic and oleic triglycerides as substrates. Insulin, condition of normolipemia and formation of hyper lipoproteinemia type IIb, IV and V

According to phylogenetic theory of general pathology, when living in ocean all were carnivorous (piscivorous) fatty acids transferring to cells in form of non-polar triglycerides nitially began apoB-48 chylomicrons, continued lipoproteins of very low and low density and fnalized its apoB-100 endocytosis. The fatty acids are transferred by chylomicrons + lipoproteins of very low density + lipoproteins of low density and non-polar triglycerides are hydrolyzed by hepatic glycerolhydrogenase and co-enzyme apoC-III; according WHO classifcation, hyperlipoproteinemia corresponds to type V. On land, in herbivorous who are not yet synthesized insulin, apoB-48 and chylomicrons left process of non-polar triglycerides transferring. In lipoproteins of very low density and lipoproteins of low density, the carnivorous transfer exogenous palmitic non-polar triglycerides. The herbivorous also transfer palmitic non-polar triglycerides though synthesized by hepatocytes from glucose endogenically. In herbivorous, transferring of palmitic non-polar triglycerides prior to synthesis of insulin is forming apoB-100 in composition of lipoproteins of very low density and lipoproteins of low density. The hydrolysis of palmitic non-polar triglycerides in lipoproteins of very low density is activated by hepatic glycerol hydrogenase and apoC-III; cells absorb lipoproteins of low density by means of apoB-100 endocytosis. The content on lipoproteins in blood plasma under electrophoresis of lipoproteins corresponds to hepatic glycerol hydrogenase type IIb. In frst and second types of fatty acids transferring in form of triglycerides to lipoproteins of very low density + lipoproteins of low density predominate palmitic fatty acid, triglycerides of the same name and palmitic metabolism of fatty acids in vivo. The insulin initiated the third type of transferring of oleic fatty acid by now to insulin-depended cells only in oleic lipoproteins of very low density; hydrolysis of oleic triglycerides is activated by late in phylogenesis post-heparin hepatic glycerol hydrogenase and apoC-II cofactor. The dynamic apoE is actively bound by apoB-100 forming apoE/B-100 ligand. At later stages of phylogenesis insulin formed fatty acids transferring in form of oleic triglycerides in lipoproteins of very low density of the same name without forming of oleic lipoproteins of low density; the electrophoregram of lipoproteins reflects absence of hepatic glycerol hydrogenase. In phylogenesis three types of fatty acids transferring to triglycerides in composition of lipoproteins formed sequentially: 1) chylomicrons + lipoproteins of very low and density + lipoproteins of low density; 2) lipoproteins of very low density + lipoproteins of low density; 3) only in lipoproteins of very low density. The frst one is specifc to piscivorous (carnivorous) while living in ocean. The second one is implemented by herbivorous while they didn't begin to synthesize insulin and hepatocytes not yet transform all endogenous palmitic fatty acid into oleic fatty acid. Insulin initiated: a) transferring of oleic fatty acids to lipoproteins of very low density without forming oleic lipoproteins of low density; b) highly effective oleic metabolism of fatty acids in vivo: c) becoming of biological function of locomotion. The aphysiological induction by substrate, surplus of palmitic fatty acids in food initiate negative alterations in composition of lipoproteins in opposite direction than in case of phylogenesis. When homo sapiens, herbivorous in phylogenesis, begins to misuse carnivorous (meat) food then instead of normolipoproteinemia in blood plasma under electrophoresis of lipoproteins one can initially detect transitory hyperlipoproteinemia type IV and then prolonged hyperlipoproteinemia type IIb. If patient factually passes on to carnivorous diet then hyperlipoproteinemia type V is developing. If content of exogenous palmitic fatty acid in food surpasses physiological capacities of its transferring in oleic triglycerides as palmitoyl-oleyl-palmitate glycerol, palmitic triglycerides as oleyl-palmitoyl-palmitate glycerol begin to form and epigenetically aphysiological non-ligand palmitic lipoproteins of very low density → lipoproteins of low density are formed. Their circulation in blood is a cause of hypertriglyceridemia, higher level of cholesterol-lipoproteins of low density, compensatory increasing of apoC-III. Then occurs induced by substrate formation of hyperlipoproteinemia initially of type IV, then of type IIb and fnally of type V. The pathogenesis of atherosclerosis and atheromotosis is activated when homo sapiens, herbivorous in phylogenesis, begin to misuse carnivorous food affecting biological functions of trophology, reaction of exotrophy (external nutrition), function of homeostasis, endoecology and function of adaptation. The formation of palmitic metabolism if fatty acids instead of oleic one is a cause of chronic defciency of energy and ATP synthesis in vivo. Insulin activates absorption of glucose by cells with purpose to use it for synthesis of oleic fatty acids. In the frst place, insulin regulates in vivo metabolism of fatty acids and only in second place metabolism of glucose.

The phylogenetic theory of general pathology. The development of seven biological functions, seven specific ones and etiological factors of metabolic epidemics, millions years' common pathogenesis and basics of prevention.

The clearer realization is coming that in general biology, medicine, historical science, in biologic phylogenetic anamnesis of Homo sapiens a task exists only one by its significance and unresolved still: to establish historical succession of becoming of specific etiologic factors of every metabolic epidemic and to understand becoming of millions years' common pathogenesis of all metabolic pandemics. The carnivorous feeding by ocean, herbivorous feeding of ancestors of rodents and Homo sapiens by dry land; misusing carnivorous food by phylogenetically herbivorous human during last centuries, disorder of biological function of trophology, biological reaction of exotrophy, pathology of biologic functions of adaptation and endoecology, features of cognitive biological function formed etiological factors of atherosclerosis and atheromatosis. The abnormalities of biological reaction metabolism↔micro-circulation; discrepancy of mechanisms of regulation of biological reaction at 2d and 3d levels of relative biological perfection in vivo, disorder of biological function of adaptation, biological reaction of compensation formed etiological factors of metabolic arterial hypertension. The synthesis of fatty acids in phylogenesis millions of years before of glucose, preference of cells to absorb fatty acids instead of glucose; previously in phylogenesis effect of hyperglycemia, glucagon and late function of insulin formed priority of metabolism of fatty acids in energetics of organisms and glucose only in the second instance. The formation in phylogenesis of methodological mode of biological subordination in functioning of humoral mediators, incapability of insulin late in phylogenesis of blocking adaptive action of humoral mediators earlier in phylogenesis, especially in visceral fatty tissue form etiological factors of resistance to insulin. The functional limitation of cognitive biological function, difficulties in maintaining unity of internal medium of organism and effects of aphysiologic factors of external medium produces necessity of completing cognitive biological function with activation of biological reaction of apoptosis and autophagy. This occurrence forms etiologic factors of metabolic syndrome - a local pathology of early, visceral, limited in numbers of cells of pool of insulin-independent visceral fatty tissue. The inconsistency of cognitive biological function, necessity of correcting it by more effective biological function of intellect, role of insulin in energy support of new biological function - locomotion, formation of unlimited in numbers of cells а pool late in phylogenesis of insulin-dependent subcutaneous adipocytes, formed etiological factors of obesity. The biological function of depositing of fatty acids in phylogenesis in vivo is characterized by etiological dependence: depositing of fatty acids occurs in the form of non-polar triglycerides and releasing into intercellular medium occurs only in form of polar unesterified fatty acids. The features of induction with substrate, formation of positioning isoforms of fatty acids, impossibility of hydrolyzing them at effecting of hormone-dependent lipase compel to eliminate aphysiologic triglycerides from liver together with hepatocytes implementing biological reactions of apoptosis and autophagy. They become etiological factors of non-alcoholic fatty disease of liver; these are aphysiologic positioning iusoforms of triglycerides as palmitoyl-palmitol-palmitate and stearyl-stearyl-stearate. The formation of endogenous hyperuricemia in phylogenesis was initiated by 2 mutations. The first one - a gene of ascorbic acid "minus" deprived Homo sapiens of synthesis of hydrophilic acceptor of active forms of oxygen. The second one - a gene of uricase "minus" blocked synthesis of allantoin but provided a possibility of re-absorbing uric acid from a pool of primary urine and use it in vivo as an acceptor of active forms of oxygen instead of ascorbic acid under realization of biological function of endoecology,biologic reaction of inflammation.

Analysis of triglyceride degradation products in drying oils and oil paints using LC–ESI-MS

An LC–ESI-MS method is presented as a novel approach for the study of aged drying oils and oil paints in various stages of oxidation and hydrolysis. The method involves separation and detection of glycerides and fatty acids on a reversed phase column using a polar gradient ranging from methanol/water to methanol/isopropanol with post-column addition of NH4Ac to facilitate electrospray ionisation. This setup allows for a reasonable separation of non-polar triglycerides in drying oil as well as very polar oxidised and hydrolysed tri, di and monoglycerides as well as free fatty acids. Detection is performed by using both positive and negative ionisation mode: positive ions for glycerides, negative ions for carboxylic acid containing degradation products and free fatty acids.In this way, distinction can be made between components in oil and metal stearate mixtures by independently probing the palmitic acid/stearic acid (P/S) ratios of the free fatty acids which mostly derive from the metal stearates, and the glycerides which derive only from the drying oil components.Analyses of 10 year-old titanium white oil paints with medium exudations and 62 year-old paints from Winsor&Newton are presented as examples to show the applicability of the method.

DIFFERENCES IN PHYLOGENETIC FACTORS OF ETIOLOGY AND PATHOGENESIS UNITY OF METABOLIC PANDEMIAS - CIVILIZATION DISEASES

Regulation of metabolism in vivo can be understood when considering the formation in phylogenesis of the function of humoral, hormonal mediators, vegetative regulators separately: a) at the cell level; b) in paracrinally regulated cell communities (PC), organs and systems and c) at the body level. Each of regulation levels was formed at different stages of phylogenesis; the completion of levels occurred, we suppose, when reaching “relative biological perfection”. Formation of organs and organ systems was finished also, in our opinion, at the level of this perfection. When three levels reached sequentially were analyzed in Homo sapiens , it was revealed with systemic approach that through “relative biological perfection” at the level of the body, “regular mismatch” of metabolism is observed. Phylogenetic mismatch of the regulation of metabolism at three levels of “relative biological perfection” are etiological factors of “metabolic pandemias” of each of “civilization diseases”. All “metabolic pandemias” including atherosclerosis, metabolic arterial hypertension, IR status, obesity, metabolic syndrome and nonalcoholic fatty liver disease are disorders of one, late in phylogenesis, biological function of locomotion. Etiological basis for metabolic (essential) arterial hypertension is a regular mismatch: a local compensation of disturbances of the biological response metabolism ↔ microcirculation in the distal arterial bed with endothelium-dependent vasodilatation and only systemic compensation at the level of the body in the proximal part. For millions of years, there coexist two functionally connected and phylogenetically separated pools of fatty cells: the earlier visceral fatty cells (VFC) and the later subcutaneous adipocytes. Insulin blocks TG hydrolysis and release of nonetherified fatty acids (NEFA) into the blood plasma only from adipocytes; hormone cannot block lipolysis in phylogenetically early VFC and inhibit the effect of phylogenetically early hormones on them. Increase in blood plasma NEFA in the course of biological response of stress in vivo does not accompany proportional growth in albumin concentration. The function of phylogenetically early resident macrophages and monocytes ↔ macrophages, differentiated in intima of arteries in situ and ex tempore , is not identical. Biological response of fatty cells deposit into adipocytes occurs in the form of nonpolar triglycerides in lipoprotein composition, but their release - only in the form of polar NEFA.

Conversion of polar and non-polar algae oil lipids to fatty acid methyl esters with solid acid catalysts – A model compound study

Bio-based fuels are becoming more and more important due to the depleting fossil resources. The production of biodiesel from algae oil is challenging compared to terrestrial vegetable oils, as algae oil consists of polar fatty acids, such as phospholipids and glycolipids, as well as non-polar triglycerides and free fatty acids common in vegetable oils. It is shown that a single sulphonated solid acid catalyst can perform the esterification and transesterification reactions of both polar and non-polar lipids. In mild reaction conditions (60–70°C) Nafion NR50 catalyst produces methyl palmitate (FAME) from the palmitic acid derivatives of di-, and tri-glyceride, free fatty acid, and phospholipid with over 80% yields, with the glycolipid derivative giving nearly 40% yields of FAME. These results demonstrate how the polar and non-polar lipid derivatives of algal oil can be utilised as feedstocks for biodiesel production with a single catalyst in one reaction step.

Foaming of Differently Processed Oats: Role of Nonpolar Lipids and Tryptophanin Proteins

ABSTRACTThe baking properties of oats are poor, mainly due to the lack of gluten matrix and hence the surface properties of the aqueous phase are crucial for the gas retention in oat dough. Our aim was to study the composition and foaming properties of the water‐soluble fraction from differently processed oats. A water extract from kilned oats contained nonpolar triglycerides and had poor foaming properties, whereas removing lipids with hexane extraction improved the foaming capacity and foam stability. A water extract from supercritical carbon dioxide extracted oats (CO2‐oats) was free from nonpolar lipids and had good foam stability and excellent foaming capacity. Moreover, oat lipid‐binding proteins, tryptophanins, were highly concentrated in the CO2‐oats‐derived foam and apparently played an important role in the foam structure. Supplementing CO2‐oats extract with small quantities (<0.05%) of nonpolar lipids of oats destructed its foaming properties. In a preliminary baking trial, the addition of the nonpolar lipids to CO2‐oats and wheat‐starch‐based baking recipes resulted in baked goods with reduced volume. The study showed that nonpolar triglycerides were present in the aqueous phase of oat in a quantity that impaired foaming. Moreover, this was the first study showing that tryptophanins, lipid‐binding proteins of oats, were highly concentrated in foams prepared of oats free of water‐extractable nonpolar lipids. In conclusion, tryptophanins can be considered as the foam‐active proteins of oats that prevent the lipid‐induced destabilization of foam structures which could improve the baking properties of oats.

Fritura de patatas en aceite de girasol. Estudio comparativo del grado de alteración del aceite de la freidora y del extraído de las patatas

The alteration ocurring in a sunflower oil used discontinuously to fry potatoes in 15 succesive occasions and in the oil content in fried potatoes was studied. According to the Perevalov colorimetric test, the spectrophotometric absorption and the percentage of both nonpolar triglycerides (%NPT) and nonpolar methyl esters (%NPME) the alteration increased either in the bath oil or in the extracted oil with the number of frying occasions. The refraction index increased only in the extracted oil. A higher thermoxidative alteration in the fried potato oil was assesed by all indexes with the exception of %NME. However, according to the %NPT such alteration was lower than the according to the %NPT such alteration was lower than the recomended for oil discarding suggesting a still acceptable quality of the fried potatoes.