LC-Orbitrap-HRMS method for analysis of traces of triacylglycerols featuring furan fatty acids.

Furan fatty acids (FuFAs) are valuable antioxidants that are highly relevant for the protection of polyunsaturated fatty acids (PUFAs) in biological systems and food. Despite their low contributions to the total fatty acids, their widespread occurrence has been documented in food and biological samples. Like other fatty acids, FuFAs are also stored esterified, e.g., in triacylglycerols. However, FuFA-containing triacylglycerols had not been detected in lipidomics analyses. Here, we present a screening method that allows for the identification of traces of FuFA-containing triacylglycerols (TAGs) utilizing LC-Orbitrap-HRMS. Initially developed with the help of purposefully synthesized FuFA-containing TAGs, the screening method was successfully applied to the analysis of two fish oil samples and one mushroom extract sample. Several FuFA-containing TAGs could be identified by direct analysis using the method and database developed in this study.

Microbial incubations of 8-phenyloctanoic acid and furan fatty acids in rumen fluid.

AIMS: The digestive tract of ruminants is specialized in the digestion of various plant components. One of the largest parts of the stomach is the so-called rumen, which contains a large number of micro-organisms that may degrade or modify fatty acids, for example by ß-oxidation, chain elongation and/or hydrogenation. METHODS AND RESULTS: Here we performed incubation experiments with less common fatty acids by in vitro incubations with rumen fluid of fistulated cows for 24 h. Sample extracts were analysed by gas chromatography with mass spectrometry. As substrates, we selected one phenyl fatty acid and four furan fatty acids (FuFAs). All studied fatty acids were degraded by ß-oxidation (two or three chain-shortening steps) while chain elongation or saturation of the aromatic part (terminal phenyl or central furan moiety) was not observed in any case. CONCLUSIONS: The percentage of ß-oxidation products was low, especially in the case of the FuFAs. This could be due to the rather long carbon number of FuFAs (19-22 carbon atoms). In addition, compound-specific differences in the degradation rates were observed in our experiments. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results produce evidence that FuFAs, which are valuable antioxidants that are known to be present in various feed items of the cow, can be effectively passed on the rumen into the milk.

Enzymatic generation and GC/MS data of triacylglycerols with furan fatty acids (FuFAs).

Furan fatty acids (FuFAs) are important minor fatty acids of food with antioxidative properties. Since their presence in food triacylglycerols (TAGs) had not been documented, we aimed to enzymatically synthesize TAGs with two important FuFAs (9M5 and 11D5). Specifically, the TAGs triolein (OOO), tripalmitin (PPP), trilinolein (LLL), tri-α-linolenin (LnLnLn), and tricaprylin (CCC) and 9M5 or 11D5 were reacted with the immobilized lipase from Candida antarctica. All reactions resulted in TAGs with one, two, and three FuFAs in decreasing order. A nomenclature system for FuFA-containing TAGs was suggested by listing FuFAs in 9M5 (11D5) style such as OO9M5 and O9M5O. Reaction of tri9M5 also resulted TAGs with 1-3 conventional fatty acids. GC/MS spectra of FuFA-containing TAGs featured two fragment ions ([M-RCO]+ and [M-RCOO]+) along with the diagnostic base peak of FuFAs and the [RCO]+ fragment ion. A data base with fragment and molecular ions of food relevant, FuFA-containing TAGs is presented as well.

Oxidative stability of tocochromanols, carotenoids, and fatty acids in maize (Zea mays L.) porridges with varying phytate concentrations during cooking and in vitro digestion.

Phytic acid, the main storage form of phosphate in maize (Zea mays L.) grains, is known as antinutrient due to its chelating properties but may also prevent oxidation. Thus, the impact of phytic acid on the degradation of tocochromanols, carotenoids, fatty acids, and oxidation products in maize during cooking and subsequent in vitro digestion was examined. Maize porridges from low phytic acid maize flour with or without admixed phytate, or from high phytic acid maize flour were prepared, and digestion experiments conducted. HPLC-(MS) or GC-MS analyses revealed a significant decrease in tocochromanols, carotenoids, and unsaturated fatty acids in the digesta compared to the maize porridges while α-tocopherylquinone and malondialdehyde concentrations increased. The addition of phytic acid did not affect the digestive stabilities of total tocochromanols and carotenoids, but increased micellarisation efficiencies of carotenoids. In conclusion, phytate did not exert antioxidant effects in maize porridge during cooking or simulated digestion.

Stable isotope analysis confirms substantial changes in the fatty acid composition of bacteria treated with antimicrobial random peptide mixtures (RPMs).

Resistance of plant-pathogenic bacteria to classic antibiotics has prompted the search for suitable alternative antimicrobial substances. One promising strategy could be the use of purposely synthesized random peptide mixtures (RPMs). Six plant-pathogenic bacteria were cultivated and treated with two RPMs previously found to show antimicrobial activity mainly by bacterial membrane disruption. Here, we show that bacteria treated with RPMs showed partly remarkable changes in the fatty acid pattern while those unaffected did not. Quantitative changes could be verified by compound specific isotope analysis of δ13C values (‰). This technique was employed due to the characteristic feature of stronger bonds between heavier isotopes in (bio)chemical reactions. As a proof of concept, the increase in abundance of a fatty acid group after RPM treatment was accompanied with a decrease in the 13C content and vice versa. We propose that our findings will help designing and synthesizing more selective antimicrobial peptides.

Detailed analysis of the fatty acid composition of six plant-pathogenic bacteria.

Bacteria show distinct and characteristic fatty acid (FA) patterns which can be modified by environmental conditions. In this study, we cultivated six plant-pathogenic bacteria of agricultural concern and performed a detailed analysis of the fatty acid composition. The study covered four strains of the gram-negative Xanthomonas campestris pathovar (pv) campestris (Xcc), Xanthomonas perforans (Xp), Acidovorax citrulli (Ac) and Pseudomonas syringae pv. tomato (Pst), and two strains of the gram-positive Clavibacter michiganensis subsp. michiganensis (Cmm) and Streptomyces scabies (Ssc). After cultivation, freeze-dried bacteria samples were transesterified and analysed by gas chromatography with mass spectrometry in full scan and selected ion monitoring (SIM) modes. Altogether, 44 different FAs were detected in the six strains with individual contributions of 0.01-43.8% to the total FAs. The variety in the six strains ranged between 12 and 31 individual FAs. The FA composition of Xcc, Xp, Cmm and Ssc were dominated by iso- and anteiso-fatty acids (especially i15:0, a15:0, i16:0), which is typical for most bacteria. In contrast to this, Ac and Pst showed only saturated and monounsaturated FAs. Four of the six bacteria showed similar FA patterns as reported before in the literature. Differences were observed in the case of Cmm where many more FAs were detected in the present study. In addition, to the best of our knowledge, the FA pattern of Xp was presented for the first time.

Location and Variety but Not Phosphate Starter Fertilization Influence the Profiles of Fatty Acids, Carotenoids, and Tocochromanols in Kernels of Modern Corn (Zea mays L.) Hybrids Cultivated in Germany.

Phosphate is a limiting plant nutrient and essential for corn growth and development. Thus, the impact of phosphate fertilization, location, and the variety of modern corn (Zea mays L.) hybrids on the profiles of fatty acids, carotenoids, and tocochromanols (vitamin E) was assessed in corn grains. Eight different corn hybrids were grown with (52.9 kg of phosphorus per ha) or without starter fertilizer at three experimental sites in Germany. Location (p < 0.05) and genetics (p < 0.001) but not phosphate fertilization significantly altered the concentrations of individual saturated and unsaturated fatty acids, carotenoids, and tocochromanols. Significant (p < 0.05) interaction effects on the concentrations were mainly observed between the variety and the location. In conclusion, the choice of the corn variety had a more significant impact on the biosynthesis of fatty acids, carotenoids, and tocochromanols than the location or phosphate application on phosphate-sufficient soils.

Characterization of single chain length chlorinated paraffin mixtures with nuclear magnetic resonance spectroscopy (NMR).

Chlorinated paraffins (CPs) are complex mixtures of polychlorinated n-alkanes of different chain length. Despite their environmental relevance, quantification is still a challenge. Moreover, the individual structures of the molecules in technical CP mixtures are largely unknown. Here, we synthesized 21 and studied 29 single chain length CP mixtures ranging from C10- to C17-CPs with different chlorine contents between 41.8% and 62.6% Cl and analyzed them by means of nuclear magnetic resonance spectroscopy (NMR). Discrete ranges of chemical shifts were observed in one-dimensional 1H and 13C NMR spectra. Two dimensional heteronuclear single quantum coherence spectroscopy (HSQC) enabled to assign nine substructures. These measurements also verified the presence of [-CCl2-] groups and chlorine atoms on terminal carbons in highly chlorinated (>59% Cl) mixtures. 1H NMR spectra of different chain length and the same degree of chlorination looked virtually the same. However, with increasing degree of chlorination the observed patterns in the spectra were slightly shifted downfield. Based on these findings, a calculation mode was developed to estimate the chlorine content (%) of the single chain length CP mixtures. The results agreed well (<0.9% mean deviation) with parallel measurements by elemental analysis.