RESUMO
Chromatographic separation of triacylglycerol (TG) enantiomers is a highly challenging task of analytical chemistry because of the similar physicochemical properties. The analysis of chiral TGs is crucial for improving the knowledge of lipid biochemistry and for understanding the nutritional properties of fats and oils. Thus, this study aimed to systematically investigate the chiral resolution of TGs consisting of three different fatty acyls (FAs). Thirty-three asymmetric TG enantiopairs, including 49 synthesized enantiopure TGs and racemic TGs, were analyzed with a recycling chiral HPLC system. Twenty-six enantiopairs were successfully separated. Overall, having both unsaturated and saturated FAs in the outer positions or a difference in carbon chain length between two saturated FAs at the outer positions favored the separation of enantiomers. The retention time at separation correlated negatively with the sn-3 carbon number of the early eluting enantiomer and positively with the carbon number difference between sn-1 and sn-3. When the samples were studied in separate groups based on unsaturation and regioisomers, both the acyl carbon number and the degree of unsaturation of FAs in all three positions influenced the separation and elution behavior of chiral TGs, indicating an active role of both intermolecular interactions and steric hindrances. This is the first systematic study of the chiral separation of TGs consisting of three different FAs using a large number of enantiopairs. The novel findings on the behavior of TG enantiomers in a chiral environment provide important guidance and reference for a stereospecific study of the chemistry and biochemistry of natural lipids.
RESUMO
The use of lipase immobilized on an octyl-agarose support to obtain the optically pure enantiomers of chiral drugs in reactions carried out in organic solvents is a great challenge for chemical and pharmaceutical sciences. Therefore, it is extremely important to develop optimal procedures to achieve a high enantioselectivity of the biocatalysts in the organic medium. Our paper describes a new approach to biocatalysis performed in an organic solvent with the use of CALB-octyl-agarose support including the application of a polypropylene reactor, an appropriate buffer for immobilization (Tris base-pH 9, 100 mM), a drying step, and then the storage of immobilized lipases in a climatic chamber or a refrigerator. An immobilized lipase B from Candida antarctica (CALB) was used in the kinetic resolution of (R,S)-flurbiprofen by enantioselective esterification with methanol, reaching a high enantiomeric excess (eep = 89.6 ± 2.0%). As part of the immobilization optimization, the influence of different buffers was investigated. The effect of the reactor material and the reaction medium on the lipase activity was also studied. Moreover, the stability of the immobilized lipases: lipase from Candida rugosa (CRL) and CALB during storage in various temperature and humidity conditions (climatic chamber and refrigerator) was tested. The application of the immobilized CALB in a polypropylene reactor allowed for receiving over 9-fold higher conversion values compared to the results achieved when conducting the reaction in a glass reactor, as well as approximately 30-fold higher conversion values in comparison with free lipase. The good stability of the CALB-octyl-agarose support was demonstrated. After 7 days of storage in a climatic chamber or refrigerator (with protection from humidity) approximately 60% higher conversion values were obtained compared to the results observed for the immobilized form that had not been stored. The new approach involving the application of the CALB-octyl-agarose support for reactions performed in organic solvents indicates a significant role of the polymer reactor material being used in achieving high catalytic activity.
Assuntos
Biocatálise , Enzimas Imobilizadas , Proteínas Fúngicas , Lipase , Sefarose , Lipase/química , Lipase/metabolismo , Enzimas Imobilizadas/química , Enzimas Imobilizadas/metabolismo , Proteínas Fúngicas/metabolismo , Proteínas Fúngicas/química , Sefarose/química , Propionatos/química , Estereoisomerismo , Cinética , Esterificação , Temperatura , Estabilidade Enzimática , Candida/enzimologia , Solventes/química , SaccharomycetalesRESUMO
This report demonstrates the first asymmetric synthesis of enantiopure structured triacylglycerols (TAGs) of the ABC type presenting three non-identical fatty acids, two of which are unsaturated. The unsaturated fatty acids included monounsaturated oleic acid (C18:1 n-9) and polyunsaturated linoleic acid (C18:2 n-6). This was accomplished by a six-step chemoenzymatic approach starting from (R)- and (S)-solketals. The highly regioselective immobilized Candida antarctica lipase (CAL-B) played a crucial role in the regiocontrol of the synthesis. The synthesis also benefited from the use of the p-methoxybenzyl (PMB) ether protective group, which enabled the incorporation of two different unsaturated fatty acids into the glycerol skeleton. The total of six such TAGs were prepared, four constituting the unsaturated fatty acids in the sn-1 and sn-2 positions, with a saturated fatty acid in the remaining sn-3 position of the glycerol backbone. In the two remaining TAGs, the different unsaturated fatty acids accommodated the sn-1 and sn-3 end positions, with the saturated fatty acid present in the sn-2 position. Enantiopure TAGs are urgently demanded as standards for the enantiospecific analysis of intact TAGs in fats and oils.
Assuntos
Ácidos Graxos , Glicerol , Éteres , Ácido Linoleico , TriglicerídeosRESUMO
The asymmetric synthesis of polyunsaturated triene C18:3 n-3 and C18:3 n-6 methoxylated ether lipids (MEL) of the 1-O-alkyl-sn-glycerol type is described as possible structural candidates for a triene C18:3 MEL of an unknown identity found in a mixture of shark and dogfish liver oil. Their C18:3 hydrocarbon chains constitute an all-cis methylene skipped n-3 or n-6 triene framework, along with a methoxyl group at the 2'-position and R-configuration of the resulting stereogenic center. The methoxylated polyenes are attached by an ether linkage to the pro-S hydroxymethyl group of the glycerol backbone. The syntheses were based on the polyacetylene approach that involves a semi-hydrogenation of the resulting triynes. Both syntheses were started from our previously described enantio- and diastereomerically pure isopropylidene-protected glyceryl glycidyl ether, a double-C3 building block that was designed as a head group synthon for the synthesis of various types of MELs.
Assuntos
Éter , Ácidos Graxos Ômega-3 , Glicerol , Etil-Éteres , Éteres , Éteres de GlicerilaRESUMO
The first total synthesis of a docosahexaenoic acid (DHA)-like methoxylated ether lipid (MEL) is reported. This compound constitutes an all-cis methylene skipped hexaene framework identical to that present in DHA, the well-known omega-3 polyunsaturated fatty acid. The polyene C22 hydrocarbon chain, bearing a methoxyl group in the 2-position and R-configuration at the resulting chiral center, is attached by an ether linkage to the pro-S hydroxymethyl group (sn-1 position) of a glycerol backbone. The asymmetric synthesis is highly convergent and based on the polyacetylene approach involving iterative copper-promoted coupling reactions of propargyl bromides with terminal alkynes and semihydrogenation of the resulting hexayne. Starting from enantiopure R-solketal and racemic epichlorohydrin, the targeted MEL was accomplished in an 8.2% yield over eight steps (longest linear sequence) involving an enantio- and diastereopure glyceryl glycidyl ether key C6-building blocks from which the polyynes were constructed.
Assuntos
Ácidos Docosa-Hexaenoicos , Éter , Éteres de Glicerila , Glicerol , Etil-ÉteresRESUMO
The report describes the preparation and use of a double-C3 building block intended as a head group synthon in the synthesis of saturated, mono-, and polyunsaturated 1-O-alkyl-sn-glycerol type methoxylated ether lipids (MELs). The resulting head piece, an enantiopure isopropylidene-protected glyceryl glycidyl ether diastereomer, was accomplished in 49% yield (max 50%) from a 1:1 diastereomeric mixture obtained from R-solketal and racemic epichlorohydrin after treatment with the Jacobsen (S,S)-Co(III)salen catalyst for the hydrolytic kinetic resolution of terminal epoxides. The diol hydrolytic product obtained in 47% yield from the unwanted diastereomer was reconverted into epoxide with an inversion of configuration in a three-step operation involving a highly regioselective lipase. This enabled the recovery of a substantial amount of diastereopure material after a subsequent treatment with the Jacobsen catalyst to furnish the oxirane head piece in altogether 72% yield of higher than 99% diastereomeric purity. A modified synthesis of a monounsaturated 16:1 MEL confirmed the correct stereochemistry and excellent enantiopurity of the head piece and resulted in a dramatic improvement in yields, efficiency, and economy of the synthesis.
Assuntos
Epicloroidrina , Glicerol , Compostos de Epóxi , Éter , Éteres de Glicerila , Lipase , LipídeosRESUMO
The shocking severity of the Covid-19 pandemic has woken up an unprecedented interest and accelerated effort of the scientific community to model and forecast epidemic spreading to find ways to control it regionally and between regions. Here we present a model that in addition to describing the dynamics of epidemic spreading with the traditional compartmental approach takes into account the social behaviour of the population distributed over a geographical region. The region to be modelled is defined as a two-dimensional grid of cells, in which each cell is weighted with the population density. In each cell a compartmental SEIRS system of delay difference equations is used to simulate the local dynamics of the disease. The infections between cells are modelled by a network of connections, which could be terrestrial, between neighbouring cells, or long range, between cities by air, road or train traffic. In addition, since people make trips without apparent reason, noise is considered to account for them to carry contagion between two randomly chosen distant cells. Hence, there is a clear separation of the parameters related to the biological characteristics of the disease from the ones that represent the spatial spread of infections due to social behaviour. We demonstrate that these parameters provide sufficient information to trace the evolution of the pandemic in different situations. In order to show the predictive power of this kind of approach we have chosen three, in a number of ways different countries, Mexico, Finland and Iceland, in which the pandemics have followed different dynamic paths. Furthermore we find that our model seems quite capable of reproducing the path of the pandemic for months with few initial data. Unlike similar models, our model shows the emergence of multiple waves in the case when the disease becomes endemic.
RESUMO
This report describes the synthesis of reversed structured 1-O-alkyl-2,3-diacyl-sn-glycerols (DAGEs) possessing a pure saturated even number fatty acid (C6:0-C16:0) at the sn-2 position along with a pure EPA or DHA located at the terminal sn-3 position of the glycerol backbone of chimyl, batyl and selachyl alcohols. These adducts were synthesized by a highly efficient two-step chemoenzymatic process involving an immobilized Candida antarctica lipase to introduce pure EPA and DHA activated as oxime esters exclusively to the sn-3 terminal position of enantiopure chimyl, batyl and selachyl alcohols in excellent yields. The saturated fatty acids were subsequently incorporated to the remaining sn-2 position of the resulting 3-monoacylglyceryl ethers (3-MAGEs) using EDAC coupling agent in the presence of DMAP in very high to excellent yields (85%-98%). No losses of enantiomeric composition were observed during these processes. The multiple utilities of the resulting focused library of reversed structured DAGEs are discussed including how such compounds may possibly be utilized within the pharmaceutical area.
Assuntos
Diglicerídeos/síntese química , Éteres Fosfolipídicos/síntese química , Espectroscopia de Ressonância Magnética , EstereoisomerismoRESUMO
SCOPE: To study the effect of positional distribution of docosahexaenoic acid (DHA) in dietary triacylglycerols (TAG) on the tissue fatty acid content and composition of mildly (n-3) deficient rats. METHODS AND RESULTS: In a 5-day feeding trial, mildly (n-3) deficient rats received 360 mg daily structured TAGs: sn-22:6(n-3)-18:0-18:0, sn-18:0-18:0-22:6(n-3), sn-18:0-22:6(n-3)-18:0, or tristearin. A fifth group receives standard (n-3) adequate feed AIN-93G from birth till the end of the trial. The DHA-fed groups show significantly higher DHA levels in the liver and visceral fat compared to the tristearin or normal feed groups showing that the dose and the short feeding period of DHA were sufficient to restore the DHA content in the organs of (n-3) deficient rats. Feeding sn-1 DHA resulted in higher levels of DHA in the liver TAG compared to sn-3 DHA feeding, although the difference did not reach statistical significance. CONCLUSION: These findings indicated a possible difference in the tissue accumulation and/or metabolic fate of DHA from the sn-1 and sn-3 positions of TAG.
Assuntos
Ácidos Docosa-Hexaenoicos , Ácidos Graxos Ômega-3 , Ratos , Animais , Ácidos Docosa-Hexaenoicos/metabolismo , Ácidos Graxos Ômega-3/metabolismo , Triglicerídeos/metabolismo , Dieta , Ácidos GraxosRESUMO
SCOPE: N-3 polyunsaturated fatty acids (n-3 PUFAs) play important roles in cognitive functions. However, there is a lack of knowledge on the metabolic impact of regio- and stereo-specific positioning of n-3 PUFAs in dietary triacylglycerols. METHODS AND RESULTS: Rats in a state of mild n-3 PUFA deficiency are fed daily with 360 mg triacylglycerols containing DHA (docosahexaenoic acid) at sn (stereospecific numbering)-1, 2, or 3 positions and 18:0 at remaining positions, or an equal amount of tristearin for 5 days. Groups fed with n-3 deficient diet and normal n-3 adequate diet are included as controls. The metabolic profiles of the brain and liver are studied using NMR (nuclear magnetic resonance)-based metabolomics. Several metabolites of significance in membrane integrity and neurotransmission, and glutamate, in particular, are significantly lower in the brain of the groups fed with sn-1 and sn-3 DHA compared to the sn-2 DHA group. Further, the tristearin and DHA groups show a lower lactate level compared to the groups fed on normal or n-3 deficient diet, suggesting a prominent role of C18:0 in regulating energy metabolism. CONCLUSION: This study sheds light on the impact of stereospecific positioning of DHA in triacylglycerols and the role of dietary stearic acid on metabolism in the brain and liver.
Assuntos
Ácidos Docosa-Hexaenoicos , Ácidos Graxos Ômega-3 , Ratos , Animais , Ácidos Docosa-Hexaenoicos/metabolismo , Ácidos Graxos Ômega-3/metabolismo , Triglicerídeos/metabolismo , Fígado/metabolismo , Encéfalo/metabolismoRESUMO
Omega-3 fatty acids such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are essential for human health but prone to oxidation. While esterification location is known to influence the stability of omega-3 in triacylglycerols (TAGs) in oxidation trials, their oxidative behavior in the gastrointestinal tract is unknown. Synthesized ABA- and AAB-type TAGs containing DHA and EPA were submitted to static in vitro digestion for the first time. Tridocosahexaenoin and DHA as ethyl esters were similarly digested. Digesta were analyzed by gas chromatography, liquid chromatography-mass spectrometry, and nuclear magnetic resonance spectroscopy. Besides the formation of di- and monoacylglycerols, degradation of hydroperoxides was detected in ABA- and AAB-type TAGs, whereas oxygenated species increased in tridocosahexaenoin. Ethyl esters were mainly unaffected. EPA was expectedly less susceptible to oxidation prior to and during the digestion process, particularly in sn-2. These results are relevant for the production of tailored omega-3 structures to be used as supplements or ingredients.
Assuntos
Ácido Eicosapentaenoico , Ácidos Graxos Ômega-3 , Humanos , Cromatografia Gasosa-Espectrometria de Massas , Ácidos Docosa-Hexaenoicos , Triglicerídeos , Digestão , ÉsteresRESUMO
Docosahexaenoic acid (DHA) is essential for health but easily oxidized. Yet the influence of DHA's exact location (sn-1, sn-2, or sn-3) in triacylglycerols on oxidative stability is currently unknown. This is the first study comparing oxidative stability of DHA in regio- and enantiopure triacylglycerols with or without RRR-α-tocopherol. Headspace solid-phase micro-extraction with gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, and nuclear magnetic resonance spectroscopy were applied. DHA in sn-2 was the most stable with or without added RRR-α-tocopherol resulting in differences in hydroperoxide formation. Without antioxidant, stability of DHA in sn-1 and sn-3 was mainly similar, with slight tendency towards better stability in sn-3. With RRR-α-tocopherol higher stability in sn-1 compared to sn-3 was observed. This points to diastereomeric interactions between RRR-α-tocopherol and DHA in sn-1. These results are highly relevant for enzymatic restructuring processes of DHA-rich fish or microalgae oil concentrates aimed for food supplements or food fortification.
Assuntos
Antioxidantes , Ácidos Docosa-Hexaenoicos , Animais , Ácidos Docosa-Hexaenoicos/análise , Triglicerídeos/química , alfa-Tocoferol , Peróxido de Hidrogênio , Estresse OxidativoRESUMO
This study investigated the impact of regio- and stereospecific position of docosahexaenoic acid (DHA) in dietary triacylglycerols (TAGs) on the fatty acid composition of tissues and organs in rats. Four-week feeding with TAGs containing DHA in sn-1, 2, or 3 position and palmitic acid in the remaining positions at a daily dosage of 500 mg TAG/kg body weight significantly increased the DHA content in all organs and tissues in rats, except in the brain, where the change in DHA level was not statistically significant. The group fed sn-1 DHA showed a significantly higher content of DHA in the plasma TAG than the group fed sn-3 DHA. The sn-3 DHA group had higher levels of DHA in the visceral fat compared to the sn-1, sn-2, as well as all other groups. This is the first study showing that DHA from sn-1 and sn-3 positions of dietary TAGs have differential accumulation in tissues. The new findings improved the current knowledge on the significance of TAG isomeric structure for the bioavailability and metabolic fate of DHA.
Assuntos
Ácidos Docosa-Hexaenoicos , Ácidos Graxos , Ratos , Animais , Triglicerídeos/química , Ácidos Docosa-Hexaenoicos/química , Dieta , Ácido PalmíticoRESUMO
The stereospecific structure of triacylglycerols (TAGs) affects the bioavailability of fatty acids. Lack of enantiopure reference compounds and effective enantiospecific methods have hindered the stereospecific analysis of individual TAGs. Twelve novel enantiostructured AAB-type TAGs were synthesized containing one of the three n-3 polyunsaturated fatty acid: α-linolenic acid (ALA), eicosapentaenoic acid (EPA), or docosahexaenoic acid (DHA) in sn-1 or sn-3 position. These compounds formed six enantiomer pairs, which were separated with recycling high-performance liquid chromatography using chiral columns and UV detection. The chromatographic retention behavior of the enantiomers and the stereospecific elution order were studied. The enantiomer with an n-3 PUFA in the sn-1 position eluted faster than the enantiomer with the n-3 PUFA in the sn-3 position, regardless of the carbon chain length and number of double bonds of the PUFA. TAG enantiomers containing DHA exhibited highly different retention on the chiral column and were separated after the first column, whereas recycling was needed to separate the enantiomer pairs containing ALA or EPA. The system using two identical columns and one mobile phase, without sample derivatization, proved to be very effective also for peak purity assessment, confirming the enantiopurity of the synthesized structured TAGs being higher than 98 % (96 % ee).
Assuntos
Ácidos Graxos Ômega-3/química , Triglicerídeos/química , Cromatografia Líquida de Alta Pressão , Estrutura Molecular , Estereoisomerismo , Triglicerídeos/síntese químicaRESUMO
Acellular fish skin (ACS) has emerged as a dermal substitute used to promote wound healing with decreased scar formation and pain relief that may be due to polyunsaturated fatty acid (PUFA) content. However, the PUFA content of ACS is still unknown. The aim of this study was to compare the total fatty acids and lipid profiles of ACS to two bovine-based grafts and standard of care human cadaver skin (HCS). Furthermore, there was also the goal to assess the capability of ACS lipid content to enhance wound healing. The fatty acid analysis was performed with GC-FID, and an LC-MS untargeted method was developed in order to the analyse the lipid profiles of the grafts was. The enhancement of wound healing by the ACS extract was investigated in vitro on HaCat cells. Our results showed that ACS had the highest content of PUFA (27.0 ± 1.43% of their total fatty acids), followed by HCS (20.6 ± 3.9%). The two grafts of bovine origin presented insignificant PUFA amounts. The majority of the PUFAs found in ACS were omega-3, and in HCS, they were omega-6. The untargeted lipidomics analysis demonstrated that ACS grafts were characterized by phosphatidylcholine containing either 20:5 or 22:6 omega-3 PUFA. The ACS lipid extract increased the HaCat cells migration and enhanced wound closure 4 hr earlier versus control. Our study demonstrated that ACS has a lipid profile that is distinct from other wound healing grafts, that PUFAs are maintained in ACS post-processing as phosphatidylcholine, and that ACS lipid content influences wound healing properties.
Assuntos
Derme Acelular , Ácidos Graxos Ômega-3/farmacologia , Pele Artificial , Cicatrização/efeitos dos fármacos , Animais , Linhagem Celular Transformada , Gadus morhua , HumanosRESUMO
Lack of synthetic enantiospecific triacylglycerols (TAGs) has hindered our understanding of the impact of TAG structure on the absorption and metabolic fate of fatty acids (FAs). In a five-day feeding trial with mildly (n-3) deficient rats, the bioavailability of docosahexaenoic acid [22:6(n-3), DHA] and stearic acid (18:0) from the two different enantiomers of TAG: sn-22:6(n-3)-18:0-18:0 and sn-18:0-18:0-22:6(n-3), and their regioisomeric TAG: sn-18:0-22:6(n-3)-18:0 was compared. Less secretion of fecal DHA was detected from the sn-2 position compared with the sn-1 and sn-3 positions, but no difference was found in DHA content of the fasting plasma or in the weight of the body or organs. 18:0 was lost to feces mainly as cleaved from the primary positions but also as glycerol-bound. The 5-day intervention in rats was long enough to modify the fatty acid profile of plasma phospholipids.
Assuntos
Ácidos Docosa-Hexaenoicos/metabolismo , Triglicerídeos/metabolismo , Animais , Disponibilidade Biológica , Peso Corporal , Dieta , Ácidos Docosa-Hexaenoicos/sangue , Ácidos Docosa-Hexaenoicos/química , Fezes/química , Masculino , Fosfolipídeos/sangue , Ratos , Ratos Sprague-Dawley , Estereoisomerismo , Triglicerídeos/químicaRESUMO
Epithelial to mesenchymal transition (EMT) is a developmental event characterized by phenotypic switching from a polarized epithelial phenotype to an unpolarized mesenchymal phenotype. Changes to plasma membrane function accompany EMT yet the differences in lipid composition of cells that have undergone EMT are relatively unexplored. To address this the lipidome of two cell models of EMT in breast epithelial tissue, D492 and HMLE, were analyzed by untargeted LC-MS. Detected masses were identified and their abundance was compared through multivariate statistical analysis. Considerable concordance was observed in eight lipid components between epithelial and mesenchymal cells in both cell models. Specifically, an increase in phosphatidylcholine and triacylglycerol were found to accompany EMT while phosphatidylcholine- and phosphatidylethanolamine plasmalogens, as well as diacylglycerols decreased. The most abundant fatty acid lengths were C16 and C18 but mesenchymal cells had on average shorter and more unsaturated fatty acids. The results are consistent with enhanced cell mobility post EMT and reflect a consequence of oxidative stress pre- and post EMT in breast epithelial tissue.
Assuntos
Transição Epitelial-Mesenquimal , Ácidos Graxos/metabolismo , Metabolismo dos Lipídeos , Glândulas Mamárias Humanas/metabolismo , Plasmalogênios/metabolismo , Células Epiteliais/citologia , Feminino , Humanos , Glândulas Mamárias Humanas/citologiaRESUMO
A fast, one-step gas chromatographic method was developed to analyze trimethylsilyl (TMS) derivatives of sugars, fruit acids, and ascorbic acid in sea buckthorn (Hippohaë rhamnoides L.) berries. The method was applied to berry press juice of sea buckthorn of different origins grown in Finland during the 2003 and 2004 seasons. The method gave reliable results for D-fructose, D-glucose, ethyl-D-glucose, and malic, quinic, and ascorbic acids, which are the major sugars and acids in sea buckthorn juice. For the first time in sea buckthorn and evidently in any berry, the presence of ethyl beta-D-glucopyranoside is reported. The structure of ethyl glucose was verified by high-performance liquid chromatography (HPLC), gas chromatography (GC), MS, and NMR analyses of both the isolated and the synthesized compounds. In the GC method, vitamin C was analyzed as ascorbic acid only, and dehydroascorbic acid was thus not taken into account.
Assuntos
Ácido Ascórbico/análise , Carboidratos/análise , Ácidos Carboxílicos/análise , Frutas/química , Hippophae/química , Ácido Quínico/análise , Cromatografia Gasosa , Finlândia , Frutose/análise , Glucose/análise , Espectroscopia de Ressonância Magnética , Malatos/análiseRESUMO
Enantiomers of racemic triacylglycerol (TAG) mixtures were separated using two chiral HPLC columns with a sample recycling system and a UV detector. A closed system without sample derivatisation enabled separation and identification by using enantiopure reference compounds of eleven racemic TAGs with C12-C22 fatty acids with 0-2 double bonds. The prolonged separation time was compensated for by fewer pretreatment steps. Presence of one saturated and one unsaturated fatty acid in the asymmetric TAG favoured the separation. Enantiomeric resolution, at the same time with stronger retention of TAGs, increased with increasing fatty acid chain length in the sn-1(3) position. Triunsaturated TAGs containing oleic, linoleic or palmitoleic acids did not separate. The elution order of enantiomers was determined by chemoenzymatically synthesised enantiopure TAGs with a co-injection method. The method is applicable to many natural fats and oils of low unsaturation level assisting advanced investigation of lipid synthesis and metabolism.