The goal of this work is the study of possibilities of two basic separation modes used in the analysis of complex triacylglycerol (TG) samples of plant oils and animal fats, i.e. non-aqueous reversed-phase (NARP) and silver-ion HPLC coupled with atmospheric pressure chemical ionization mass spectrometry (APCI-MS). The orthogonality of both separation modes is tested for complex TG mixtures containing fatty acids (FAs) with different acyl chain lengths, different number, positions and geometry of double bonds (DBs) and different regioisomeric positions of FAs on the glycerol skeleton. The retention in NARP mode is governed by the equivalent carbon number, while the retention in silver-ion chromatography increases with the increasing number of DBs with a clear differentiation between cis- and trans-FAs. Moreover, silver-ion mode enables at least the partial resolution of regioisomeric TG mixtures including cis-/trans-regioisomers, as illustrated on two examples of randomization mixtures. Off-line 2D coupling of both complementary modes (NARP in the first dimension and silver-ion in the second dimension) yields the superior chromatographic selectivity resulting in the highest number of identified TGs ever reported for studied samples. Off-line 2D chromatograms are processed with the home-made software providing various ways of data visualization.

• MeSH
• hmotnostní spektrometrie metody   MeSH
• oleje rostlin chemie   MeSH
• stříbro chemie   MeSH
• triglyceridy analýza   MeSH
• vysokoúčinná kapalinová chromatografie metody   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• oleje rostlin MeSH
• stříbro MeSH
• triglyceridy MeSH

Enantiomers of triacylglycerols (TAGs) containing any combination of very long chain fatty acids (VLCFAs) and/or very long chain polyunsaturated fatty acids (VLCPUFAs) with diolein, dilinolein and didocosahexaenoin were synthesized. Gradient non-aqueous reversed-phase high-performance liquid chromatography/high resolution atmospheric pressure chemical ionization-tandem mass spectrometry (NARP-HPLC/HRMS2-APCI) and chiral liquid chromatography were used for the separation and identification of molecular species of these TAGs. Further, NARP-LC and chiral LC were used to separate natural mixtures of TAGs obtained from four natural sources, i.e. ximenia oil (Ximenia americana), green alga (Botryococcus braunii), breweŕs yeast (Saccharomyces pastorianus) and a dinoflagellate (Amphidinium carterae). The ratio of regioisomers and enantiomers in individual samples was determined and a hypothesis has been confirmed on the biosynthetic pathway of natural TAGs, which is based on the preferential representation of VLCFAs and VLCPUFAs in the sn-1 position of the glycerol backbone.

• Klíčová slova
• Enantiomeric separation, Non-aqueous reversed-phase-HPLC, Triacylglycerols, Very long chain fatty acids, Very long-chain polyunsaturated fatty acids,
• MeSH
• atmosférický tlak MeSH
• Chlorophyta chemie   metabolismus   MeSH
• chromatografie s reverzní fází MeSH
• Dinoflagellata chemie   metabolismus   MeSH
• kyseliny mastné neesterifikované chemie   MeSH
• nenasycené mastné kyseliny chemie   MeSH
• Saccharomyces chemie   metabolismus   MeSH
• stereoizomerie MeSH
• tandemová hmotnostní spektrometrie MeSH
• triglyceridy analýza   chemie   izolace a purifikace   MeSH
• vysokoúčinná kapalinová chromatografie MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• kyseliny mastné neesterifikované MeSH
• nenasycené mastné kyseliny MeSH
• triglyceridy MeSH

Edible conifer seeds can serve as a source of triacylglycerols (TGs) with unusual Delta5 unsaturated polymethylene interrupted fatty acids (UPIFAs), such as cis-5,9-octadecadienoic (taxoleic), cis-5,9,12-octadecatrienoic (pinolenic), cis-5,11-eicosadienoic (keteleeronic) and cis-5,11,14-eicosatrienoic acids (sciadonic). Conifer seed oils from European Larch (Larix decidua), Norway Spruce (Picea abies) and European Silver Fir (Abies alba) have been analyzed by non-aqueous reversed-phase high-performance liquid chromatography (NARP-HPLC) with atmospheric pressure chemical ionisation (APCI)-MS detection. The influence of different positions of double bonds in Delta5-UPIFAs on the retention and fragmentation behavior is described and used for the successful identification of TGs in each oil. TGs containing Delta5-UPIFAs have a higher retention in comparison with common TGs found in plant oils with single methylene interrupted Delta6(9)-FAs and also significantly changed relative abundances of fragment ions in APCI mass spectra. Results obtained from HPLC/MS analyses are supported by validated GC/FID analyses of fatty acid methyl esters after the transesterification. The total content of Delta5-UPIFAs is about 32% for European Larch, 27% for Norway Spruce and 20% for European Silver Fir. In total, 20 FAs with acyl chain lengths from 16 to 24 carbon atoms and from 0 to 3 double bonds have been identified in 64 triacylglycerols from 3 conifer seed oils.

• MeSH
• atmosférický tlak MeSH
• chromatografie plynová přístrojové vybavení   metody   MeSH
• hmotnostní spektrometrie s elektrosprejovou ionizací přístrojové vybavení   metody   MeSH
• modřín chemie   MeSH
• nenasycené mastné kyseliny analýza   MeSH
• oleje rostlin chemie   MeSH
• plamínková ionizace MeSH
• reprodukovatelnost výsledků MeSH
• semena rostlinná chemie   MeSH
• terminologie jako téma MeSH
• triglyceridy chemie   MeSH
• vysokoúčinná kapalinová chromatografie přístrojové vybavení   metody   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• delta5 polyunsaturated fatty acid MeSH Prohlížeč
• nenasycené mastné kyseliny MeSH
• oleje rostlin MeSH
• triglyceridy MeSH

Three dimensions of chromatographic separation, using split-flow two-dimensional liquid chromatography (SF-2D-LC) with two parallel second dimensions, LC × 2LC, combined with quadruple parallel mass spectrometry (LC3MS4) is demonstrated for analysis of NIST SRM 1849a adult/infant formula. The first dimension, 1D, was a conventional non-aqueous reversed-phase (NARP) HPLC separation using two C18 columns in series, followed by detection using an ultraviolet (UV) detector, a fluorescence detector (FLD), with flow then split to a corona charged aerosol detector (CAD), and then dual parallel mass spectrometry (MS), conducted in atmospheric pressure photoionization (APPI) and electrospray ionization (ESI) modes. The first second dimension, 2D(1), UHPLC was conducted on a 50.0 mm C30 column using a NARP-UHPLC parallel gradient for separation of short-chain triacylglycerols (TAGs) from long-chain TAGs, with detection by UV and ESI-MS. The second dimension, 2D(2), UHPLC was conducted using a 100.0 mm C30 column with a NARP-UHPLC parallel gradient for improved separation of TAG isomers, with detection by UV, an evaporative light scattering detector, and high-resolution, accurate-mass (HRAM) ESI-MS. Transferred eluent dilution was used to refocus peaks and keep them sharp during elution in both 2Ds. The separation space in the 2D(2) was optimized using multi-cycle (aka, "constructive wraparound") elution, which employed flow rate programming. In the 1D, calibration lines for quantification of fat-soluble vitamins were constructed. A lipidomics approach to TAG identification and quantification by HRAM-ESI-MS was applied to the 2D(2). These experiments can be represented: LC1MS2 × (LC1MS1 + LC1MS1) = LC3MS4, or three-dimensional liquid chromatography with quadruple parallel mass spectrometry.

• Klíčová slova
• 2D-LC, 3D-LC, APPI-MS, ESI-MS, Lipidomics, Two-dimensional LC,
• MeSH
• dospělí MeSH
• hmotnostní spektrometrie s elektrosprejovou ionizací * MeSH
• lidé MeSH
• náhražky mateřského mléka * MeSH
• triglyceridy MeSH
• vysokoúčinná kapalinová chromatografie MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• triglyceridy MeSH

Lipid-like compounds containing a dimethylarsinoyl group, i.e. Me2As(O)-, have been identified by liquid chromatography/inductively coupled plasma mass spectrometry (LC/ICP-MS) and non-aqueous reversed-phase high-performance liquid chromatography (positive and/or negative high-resolution tandem electrospray ionization mass spectrometry (NARP-HPLC/HR-ESI+(-)-MS/MS) from three strains of green algae of the genus Coccomyxa (Trebouxiophyceae, Chlorophyta). The algae were cultivated in a medium containing 10 g arsenic/L, i.e. 133.5 mmol/L of Na2HAsO4.7H2O. After extraction by methyl-tert-butyl ether (MTBE), total lipids were analyzed by ICP-MS or ESI-MS without any further separation or fractionation. A total of 39 molecular species of arsenic triacylglycerols (AsTAG), 15 arsenic phosphatidylcholines (AsPC), 8 arsenic phosphatidylethanolamines (AsPE), 6 arsenic phosphatidylinositols (AsPI), 2 arsenic phosphatidylglycerols (AsPG) and 5 unknown lipids (probably ceramides) were identified. The structures of all molecular species were confirmed by tandem MS. Dry matter of the individual strains contained different amounts of total arsenolipids, i.e. C. elongata CCALA 427 (0.32 mg/g), C. onubensis (1.48 mg/g), C. elongata S3 (2.13 mg/g). On the other hand, there were only slight differences between strains in the relative abundances of individual molecular species. Possible biosynthesis of long-chain lipids with the end group Me2As(O) has also been suggested.

• Klíčová slova
• Arsenolipids, Coccomyxa elongata, Coccomyxa onubensis, Dimethylarsinoyl long chains, Green alga, Inductively coupled plasma mass spectrometry (ICP-MS), Non-aqueous reversed-phase high-performance liquid chromatography (NARP-HPLC), Positive/negative high-resolution tandem electrospray ionization mass spectrometry (HR-ESI-MS/MS), Trebouxiophyceae,
• MeSH
• arsenikové přípravky chemie   izolace a purifikace   metabolismus   MeSH
• Chlorella chemie   metabolismus   MeSH
• lipidy chemie   izolace a purifikace   MeSH
• molekulární struktura MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• arsenikové přípravky MeSH
• lipidy MeSH

Non-aqueous reversed-phase high-performance liquid chromatography (NARP-HPLC) with atmospheric pressure chemical ionization (APCI) was used for separation of triacylglycerols from five strains of haptophyte algae (genera Coccolithophora, Isochrysis, and Prymnesium). This study describes the separation and identification of C18 polyunsaturated triacylglycerols containing stearidonic and octadecapentaenoic fatty acids, including their regioisomers. Salinity affects the proportion of saturated and unsaturated fatty acids. The biosynthesis of C18 polyunsaturated triacylglycerols was found to be very stereospecific and to depend on the salinity of cultivation media, asymmetric regioisomers predominating at low salinity (sn-OpOpSt and/or PoStSt) and symmetric ones at high salinity (sn-OpStOp and or StPoSt).

• Klíčová slova
• Coccolithophyceae, Haptophyte algae (genera Coccolithophora, Isochrysis and Prymnesium), Liquid chromatography-mass spectrometry, Octadecapentaenoic acid, Salinity, Stearidonic acid,
• MeSH
• Haptophyta chemie   genetika   MeSH
• kyseliny mastné omega-3 chemie   MeSH
• mastné kyseliny analýza   MeSH
• molekulární struktura MeSH
• nenasycené mastné kyseliny chemie   MeSH
• salinita MeSH
• stereoizomerie MeSH
• triglyceridy analýza   MeSH
• vysokoúčinná kapalinová chromatografie MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Německo MeSH
• Názvy látek
• kyseliny mastné omega-3 MeSH
• mastné kyseliny MeSH
• nenasycené mastné kyseliny MeSH
• stearidonic acid MeSH Prohlížeč
• triglyceridy MeSH

Quantitative analysis of triacylglycerols (TGs) in plant oils and animal fats by normalization of peak areas can lead to erroneous results due to the large response differences with common HPLC detectors between the various TGs. The charged aerosol detector (CAD), that generates an almost universal response for non-volatile compounds, was combined with non-aqueous reversed-phase HPLC (NARP-HPLC) to develop a simple quantitative method, without need for RFs, for the analysis of complex natural TG mixtures from plant oils. Two 25 cm Hypersil ODS columns, connected in series, and a mobile phase gradient composed of acetonitrile, 2-propanol and hexane were used. Mobile phase compensation was applied, by mixing of the column effluent with the inversed gradient delivered by a second HPLC pump, for the suppression of the response dependency of the analytes on the mobile phase composition. Calibration curves of 16 saturated (from C7:0 to C22:0) and 3 unsaturated (C18:1, C18:2, C18:3) single-acid TG standards were measured and their RFs were compared with a previously described method using atmospheric pressure chemical ionization-mass spectrometry (APCI-MS). The variation in response of the most common TGs (containing fatty acid chains from 12 to 19 carbons) could be reduced to less than 5% making the combination of NARP-HPLC with CAD and mobile phase compensation an adequate tool for fast quantitative analysis of TGs in common plant oils.

• MeSH
• aerosoly * MeSH
• kalibrace MeSH
• oleje rostlin chemie   MeSH
• referenční standardy MeSH
• reprodukovatelnost výsledků MeSH
• senzitivita a specificita MeSH
• spektrofotometrie ultrafialová MeSH
• triglyceridy analýza   MeSH
• vysokoúčinná kapalinová chromatografie metody   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• aerosoly * MeSH
• oleje rostlin MeSH
• triglyceridy MeSH