Analysis of glycerol bound ω-oxo-fatty acids as ω-dioxane-FAME-derivatives.

ω-oxo-fatty acids, also known as aldehydic fatty acids, are major products of fatty acid oxidation and pose potential health risks. When bound to glycerol, ω-oxo-fatty acids (core aldehydes) can be ingested with food. Challenges in GC-MS quantification include the absence of an appropriate internal standard. Additionally, substantial analyte losses during sample preparation, caused by the high volatility of short-chain compounds, alter their pattern based on molecular weight. In this study, among various tested derivatization methods, the formation of ω-dioxane derivatives demonstrated improved recovery rates after three evaporation cycles. For methyl 7-oxo-heptanoate, recovery increased from 43 % to 88 %, while recovery rates for different chain lengths and a novel synthesized internal standard improved from a range of 43 %-76 % to 87 %-92 %. Additionally, ω-dioxane derivatives displayed favorable GC-MS behavior, enabling clear identification and increased sensitivity. Finally, ω-oxo-fatty acids were quantified as their ω-dioxane-derivatives in thermally treated sunflower and rapeseed oil.

Can the AI tools ChatGPT and Bard generate energy, macro- and micro-nutrient sufficient meal plans for different dietary patterns?

Artificial intelligence chatbots based on large language models have recently emerged as an alternative to traditional online searches and are also entering the nutrition space. In this study, we wanted to investigate whether the artificial intelligence chatbots ChatGPT and Bard (now Gemini) can create meal plans that meet the dietary reference intake (DRI) for different dietary patterns. We further hypothesized that nutritional adequacy could be improved by modifying the prompts used. Meal plans were generated by 3 accounts for different dietary patterns (omnivorous, vegetarian, and vegan) using 2 distinct prompts resulting in 108 meal plans total. The nutrient content of the plans was subsequently analyzed and compared to the DRIs. On average, the meal plans contained less energy and carbohydrates but mostly exceeded the DRI for protein. Vitamin D and fluoride fell below the DRI for all plans, whereas only the vegan plans contained insufficient vitamin B12. ChatGPT suggested using vitamin B12 supplements in 5 of 18 instances, whereas Bard never recommended supplements. There were no significant differences between the prompts or the tools. Although the meal plans generated by ChatGPT and Bard met most DRIs, there were some exceptions, particularly for vegan diets. These tools maybe useful for individuals looking for general dietary inspiration, but they should not be relied on to create nutritionally adequate meal plans, especially for individuals with restrictive dietary needs.

An overview of food lipids toward food lipidomics.

Increasing evidence regarding lipids' beneficial effects on human health has changed the common perception of consumers and dietary officials about the role(s) of food lipids in a healthy diet. However, lipids are a wide group of molecules with specific nutritional and bioactive properties. To understand their true nutritional and functional value, robust methods are needed for accurate identification and quantification. Specific analytical strategies are crucial to target specific classes, especially the ones present in trace amounts. Finding a unique and comprehensive methodology to cover the full lipidome of each foodstuff is still a challenge. This review presents an overview of the lipids nutritionally relevant in foods and new trends in food lipid analysis for each type/class of lipids. Food lipid classes are described following the LipidMaps classification, fatty acids, endocannabinoids, waxes, C8 compounds, glycerophospholipids, glycerolipids (i.e., glycolipids, betaine lipids, and triglycerides), sphingolipids, sterols, sercosterols (vitamin D), isoprenoids (i.e., carotenoids and retinoids (vitamin A)), quinones (i.e., coenzyme Q, vitamin K, and vitamin E), terpenes, oxidized lipids, and oxylipin are highlighted. The uniqueness of each food group: oil-, protein-, and starch-rich, as well as marine foods, fruits, and vegetables (water-rich) regarding its lipid composition, is included. The effect of cooking, food processing, and storage, in addition to the importance of lipidomics in food quality and authenticity, are also discussed. A critical review of challenges and future trends of the analytical approaches and computational methods in global food lipidomics as the basis to increase consumer awareness of the significant role of lipids in food quality and food security worldwide is presented.

Determination of Polar Lipids in Wheat and Oat by a Complementary Approach of Hydrophilic Interaction Liquid Chromatography and Reversed-Phase High-Performance Liquid Chromatography Hyphenated with High-Resolution Mass Spectrometry.

Cereals contain lipids that fulfill important physiological roles and are associated with stress in the plant. However, many of the specific biological roles of lipids are yet unknown. Comprehensive analysis of these polar lipid categories in whole grain wheat and oat, cereals highly relevant also in nutrition, was performed. Hydrophilic interaction liquid chromatography (HILIC) and reversed-phase high-performance liquid chromatography (RP-HPLC) coupled with high-resolution mass spectrometry using electrospray ionization in both positive and negative ionization mode was used. Exploiting the different separation mechanisms, HILIC was used as a screening method for straightforward lipid class assignment and enabled differentiation of isomeric lipid classes, like phosphatidylethanolamine and lyso-N-acylphosphatidylethanolamine, while RP-HPLC facilitated separation of constitutional isomers. In combination with data-dependent MS/MS experiments, 67 lipid species belonging to nine polar lipid classes could be identified. Furthermore, with both ionization modes, fatty acyl chains directly connected to the lipid headgroups could be assigned. This work focused on the four lipid classes N-acylphosphatidylethanolamines, acyl-monogalactosyldiacylglycerols, digalactosyldiacylglycerols, and monogalactosyldiacylglycerols as they were less studied in detail in the past. Applying the complementary approach, the relative lipid species compositions in these lipid classes was investigated in detail.

Neolithic culinary traditions revealed by cereal, milk and meat lipids in pottery from Scottish crannogs.

Cereal cultivation in Britain dates back to ca. 4000 BCE, probably introduced by migrant farmers from continental Europe. Widespread evidence for livestock appears in the archaeozoological record, also reflected by ubiquitous dairy lipids in pottery organic residues. However, despite archaeobotanical evidence for domesticated plants (such as cereals), organic residue evidence has been near-absent. Our approach, targeting low-abundance cereal-specific markers, has now revealed evidence for cereals (indicating wheat) in Neolithic pottery from Scottish 'crannogs', dating to ca. 3600 - 3300 BCE. Their association with dairy products suggests cereals may have been regularly prepared together as a milk-based gruel. We also observed a strong association between the occurrence of dairy products and smaller-mouthed vessels. Here, we demonstrate that cereal-specific markers can survive in cooking pots for millennia, revealing the consumption of specific cereals (wheat) that are virtually absent from the archaeobotanical record for this region and illuminating culinary traditions among early farming communities.

Correction: The dietary impact of the Norman Conquest: A multiproxy archaeological investigation of Oxford, UK.

[This corrects the article DOI: 10.1371/journal.pone.0235005.].

Interpreting ancient food practices: stable isotope and molecular analyses of visible and absorbed residues from a year-long cooking experiment.

Chemical analyses of carbonized and absorbed organic residues from archaeological ceramic cooking vessels can provide a unique window into the culinary cultures of ancient people, resource use, and environmental effects by identifying ingredients used in ancient meals. However, it remains uncertain whether recovered organic residues represent only the final foodstuffs prepared or are the accumulation of various cooking events within the same vessel. To assess this, we cooked seven mixtures of C3 and C4 foodstuffs in unglazed pots once per week for one year, then changed recipes between pots for the final cooking events. We conducted bulk stable-isotope analysis and lipid residue analysis on the charred food macro-remains, carbonized thin layer organic patina residues and absorbed lipids over the course of the experiment. Our results indicate that: (1) the composition of charred macro-remains represent the final foodstuffs cooked within vessels, (2) thin-layer patina residues represent a mixture of previous cooking events with bias towards the final product(s) cooked in the pot, and (3) absorbed lipid residues are developed over a number of cooking events and are replaced slowly over time, with little evidence of the final recipe ingredients.

The dietary impact of the Norman Conquest: A multiproxy archaeological investigation of Oxford, UK.

Archaeology has yet to capitalise on the opportunities offered by bioarchaeological approaches to examine the impact of the 11th-century AD Norman Conquest of England. This study utilises an integrated multiproxy analytical approach to identify and explain changes and continuities in diet and foodways between the 10th and 13th centuries in the city of Oxford, UK. The integration of organic residue analysis of ceramics, carbon (δ13C) and nitrogen (δ15N) isotope analysis of human and animal bones, incremental analysis of δ13C and δ15N from human tooth dentine and palaeopathological analysis of human skeletal remains has revealed a broad pattern of increasing intensification and marketisation across various areas of economic practice, with a much lesser and more short-term impact of the Conquest on everyday lifestyles than is suggested by documentary sources. Nonetheless, isotope data indicate short-term periods of instability, particularly food insecurity, did impact individuals. Evidence of preferences for certain foodstuffs and cooking techniques documented among the elite classes were also observed among lower-status townspeople, suggesting that Anglo-Norman fashions could be adopted across the social spectrum. This study demonstrates the potential for future archaeological research to generate more nuanced understanding of the cultural impact of the Norman Conquest of England, while showcasing a method which can be used to elucidate the undocumented, everyday implications of other large-scale political events on non-elites.

Mechanisms of lipid preservation in archaeological clay ceramics revealed by mass spectrometry imaging.

Traces of lipids, absorbed and preserved for millennia within the inorganic matrix of ceramic vessels, act as molecular fossils and provide manifold information about past people's subsistence, diet, and rituals. It is widely assumed that lipids become preserved after adsorption into nano- to micrometer-sized pores, but to this day the distribution of these lipids in the ceramics was virtually unknown, which severely limits our understanding about the process of lipid preservation. Here we use secondary ion mass spectrometry (SIMS) imaging for direct in situ analysis of lipids absorbed in 700- to 2,000-y-old archaeological pottery. After sectioning from larger sherds, wall cross-sections of smaller fragments were used for SIMS analysis. Lipids were found in relatively large zones of 5- to 400-µm diameter, which does not support the notion of absorption only into individual nanometer-scale pores but indicates that more macroscopic structures in the ceramics are involved in lipid preservation as well. Furthermore, lipids were found concentrated on calcium carbonate inclusions in the ceramics, which suggests that precipitation of fatty acids as calcium salts is an important aspect of lipid preservation in archaeological samples. This has important implications for analytical methods based on extraction of lipids from archaeological ceramics and needs to be considered to maximize the yield and available information from each unique sample.

Digging deeper - A new data mining workflow for improved processing and interpretation of high resolution GC-Q-TOF MS data in archaeological research.

Gas chromatography-mass spectrometry profiling is the most established method for the analysis of organic residues, particularly lipids, from archaeological contexts. This technique allows the decryption of hidden chemical information associated with archaeological artefacts, such as ceramic pottery fragments. The molecular and isotopic compositions of such residues can be used to reconstruct past resource use, and hence address major questions relating to patterns of subsistence, diet and ritual practices in the past. A targeted data analysis approach, based on previous findings reported in the literature is common but greatly depends on the investigator's prior knowledge of specific compound classes and their mass spectrometric behaviour, and poses the risk of missing unknown, potentially diagnostic compounds. Organic residues from post-prehistoric archaeological samples often lead to highly complex chromatograms, which makes manual chromatogram inspection very tedious and time consuming, especially for large datasets. This poses a significant limitation regarding the scale and interpretative scopes of such projects. Therefore, we have developed a non-targeted data mining workflow to extract a higher number of known and unknown compounds from the raw data to reduce investigator's bias and to vastly accelerate overall analysis time. The workflow covers all steps from raw data handling, feature selection, and compound identification up to statistical interpretation.

Development of equivalent chain length (ECL) rules for lipid compounds.

Lipid compounds (fatty acids, tocochromanols, phytosterols) are difficult to separate by countercurrent chromatography (CCC) due to the existence of many similar structures and the limited availability of suitable biphasic solvent systems. Here we show that for these compound classes the success of a CCC separation can be directly derived from the chemical structures without the necessity of experimental determinations of K values. In most cases, lipid compounds differ in the total carbon number and the number of double bonds. For each structure the so-called equivalent chain length (ECL) can be calculated by subtracting a distinct value for each double bond from the total carbon number. Empirically, we verified that in the case of unbranched fatty acids (determined as methyl esters) one double bond corresponds with two carbons. Evaluation of CCC data from seventeen phytosterols in five plant oils and nine tocochromanol standards showed that one double bond was equal with one carbon for both lipid classes. Most compounds with different ECL can be separated by CCC but not those with the same ECL. In these cases, the selection of a suitable source for isolation of a particular lipid compound becomes very important. Knowledge of the impact of double bonds may also be a helpful tool for other substance classes.

Lipid profiling and analytical discrimination of seven cereals using high temperature gas chromatography coupled to high resolution quadrupole time-of-flight mass spectrometry.

Minor lipids in cereals (such as phytosterols and alkylresorcinols) can be important for human nutrition and/or be used as biomarkers for cereal intake. However, the analysis of cereal lipids is very challenging due to the complex lipidome comprising several hundred individual compounds present over a wide range of concentrations. Here we present a method for the profiling of cereal lipids using high temperature gas chromatography coupled to high resolution mass spectrometry (GC/Q-TOF MS). The method was used to investigate the lipid profiles of 77 samples of bread wheat, spelt, einkorn, emmer, barley, rye and oats. Distinct differences in the patterns of alkylresorcinols, free and conjugated sterols and tocopherols between the cereals could be observed. Furthermore, traces of tocomonoenols and diunsaturated and methyl-alkylresorcinols (not previously reported in cereals) could be detected. Finally, the lipid patterns in the cereals could be used to separate the cereals by Principal Component Analysis.

Ancient Biomolecules and Evolutionary Inference.

Over the past three decades, studies of ancient biomolecules-particularly ancient DNA, proteins, and lipids-have revolutionized our understanding of evolutionary history. Though initially fraught with many challenges, today the field stands on firm foundations. Researchers now successfully retrieve nucleotide and amino acid sequences, as well as lipid signatures, from progressively older samples, originating from geographic areas and depositional environments that, until recently, were regarded as hostile to long-term preservation of biomolecules. Sampling frequencies and the spatial and temporal scope of studies have also increased markedly, and with them the size and quality of the data sets generated. This progress has been made possible by continuous technical innovations in analytical methods, enhanced criteria for the selection of ancient samples, integrated experimental methods, and advanced computational approaches. Here, we discuss the history and current state of ancient biomolecule research, its applications to evolutionary inference, and future directions for this young and exciting field.

Countercurrent chromatographic isolation and purification of 11'-α-tocomonoenol from the vitamin E extract of palm oil.

A new vitamin E homologue, α-tocomonoenol was detected in palm oil, but was not isolated in large amounts and with high purity so far. Here we present an easy and fast method to isolate α-tocomonoenol from vitamin E rich nutrient capsules with countercurrent chromatography (CCC). With the solvent system n-hexane - benzotrifluoride - acetonitrile (10:3.5:6.5, v/v/v) about 30 mg α-tocomonoenol with a purity of 75% could be enriched in one step from 1 g crude sample. Column chromatography with 20% deactivated silica gel and n-hexane - ethyl acetate (95:5, v/v) was performed to gain 5.6 mg α-tocomonoenol with a purity of 99.5% according to GC/MS. Structural verification by 1H NMR spectroscopy verified that the double bond was located in 11'-position (11'-α-tocomonoenol). The trace impurity detected in the isolate was identified to be 12'-α-tocomonoenol, a compound previously detected in marine samples.

Tocopherols, Tocomonoenols, and Tocotrienols in Oils of Costa Rican Palm Fruits: A Comparison between Six Varieties and Chemical versus Mechanical Extraction.

Palm oil is one of the richest sources of tocotrienols and may contain other non-tocopherol vitamin E congeners. The vitamin E profiles of fully ripened fruit mesocarp of three Elaeis guineensis, two Elaeis oleifera, and one hybrid O × G palm fruit genotypes from Costa Rica were analyzed by high-performance liquid chromatography with fluorescence detection and gas chromatography-mass spectrometry after mechanical extraction by a screw press and chemical extraction with hexane. γ-Tocotrienol, α-tocotrienol, and α-tocopherol were the most abundant tocochromanols, while other tocopherols (ß-tocopherol, γ-tocopherol, and δ-tocopherol) and α-tocomonoenol were detected at minor concentrations. Significant differences in vitamin E profiles between genotypes were observed, and the variety E. oleifera Quepos (CB9204) had by far the highest content of total tocotrienols (890 µg/g of oil) and total vitamin E (892 µg/g of oil). Chemical extraction with hexane afforded up to 2.5-fold higher vitamin E yields than screw press extraction. α-Tocomonoenol co-eluted with γ-tocopherol in reversed-phase high-performance liquid chromatography analyses and is a possible source of error in the quantification of γ-tocopherol in foods.

The use of countercurrent chromatography in the separation of nonpolar lipid compounds.

Isolation of lipophilic compounds by countercurrent chromatography (CCC) is a challenge because biphasic solvent systems in which these compounds distribute evenly are difficult to obtain. In this article we present novel applications of lipid compound isolation from natural sources. Conjugated linolenic acids (CLnAs, log KOW ∼7) were isolated from pomegranate oil using a solvent system consisting of n-heptane/methanol/water 100:91:9 (v/v/v). The CLnA fraction was free of other fatty acids but consisted of different isomers which were not resolved from each other. In the less polar range (log KOW ∼12), three tocotrienols (α-, γ- and δ-tocotrienol) were isolated from a vitamin E capsule produced from palm oil by using the solvent system n-hexane/acetonitrile/benzotrifluoride (BTF) at a ratio of 10:6.5:3.5 (v/v/v). Between 36 and 65mg of each of the three tocotrienols were obtained in one injection with purities >97%. Advantages and disadvantages of the "BTF system" are discussed by comparing the phase composition with the simple n-hexane/acetonitrile system and by the fractionation of phytosterols (log KOW ∼9.5) from rapeseed oil.

Nutritional value of duckweeds (Lemnaceae) as human food.

Duckweeds have been consumed as human food since long. Species of the duckweed genera, Spirodela, Landoltia, Lemna, Wolffiella and Wolffia were analysed for protein, fat, and starch contents as well as their amino acid and fatty acid distribution. Protein content spanned from 20% to 35%, fat from 4% to 7%, and starch from 4% to 10% per dry weight. Interestingly, the amino acid distributions are close to the WHO recommendations, having e.g. 4.8% Lys, 2.7% Met+Cys, and 7.7% Phe+Tyr. The content of polyunsaturated fatty acids was between 48 and 71% and the high content of n3 fatty acids resulted in a favourable n6/n3 ratio of 0.5 or less. The phytosterol content in the fastest growing angiosperm, W. microscopica, was 50mgg(-1) lipid. However, the content of trace elements can be adjusted by cultivation conditions. Accordingly, W. hyalina and W. microscopica are recommended for human nutrition.

More than 170 polyunsaturated tocopherol-related compounds in a vitamin E capsule: Countercurrent chromatographic enrichment, gas chromatography/mass spectrometry analysis and preliminary identification of the potential artefacts.

Tocopherols and tocotrienols (usually summed up as vitamin E) are a class of structurally related natural antioxidants. Commonly, only some of the eight classic representatives (four tocopherols and four tocotrienols) are found with varied composition in food. In this study we fractionated 230mg oil from commercial vitamin E supplement capsules by countercurrent chromatography (CCC) and subsequent analysis by gas chromatography with mass spectrometry (GC/MS) of silylated CCC fractions showed that these eight isomers represented only about 70% of total tocopherol compounds. Detailed analysis enabled the detection of 161T3 isomers (α-, γ- and δ-T3) along with 18 tetra- and several penta-unsaturated isomers (tocools), two tocomonoenol isomers, and several degradation products with shorter isoprenoid side chain (apo-tocools). Altogether, over 170 tocool compounds, most likely artefacts which originated from an inappropriate oil refining process were described in this study. Silver ion high performance liquid chromatography (Ag+-HPLC) was used to separate one fraction rich in γ-T3 into four peaks each consisting of at least five peaks according to GC/MS. About ten γ-T3 isomers were also detected in rice bran oils from one producer bought retail in Germany.

Phytyl Fatty Acid Esters in the Pulp of Bell Pepper (Capsicum annuum).

Phytyl fatty acid esters (PFAE) are esters of fatty acids with the isoprenoid alcohol phytol (3,7R,11R,15-tetramethylhexadec-2E-enol). In this study, PFAE were identified and quantified in bell pepper using gas chromatography with mass spectrometry (GC-MS). All red (n = 14) and yellow (n = 6) samples contained six or seven PFAE at 0.9-11.2 mg/100 g fresh weight. By contrast, PFAE were not detected in green bell pepper samples (n = 3). PFAE might eventually be a source for bioavailable phytol, which can be transformed into phytanic acid by humans. Phytanic acid cannot be properly degraded by patients who suffer from Refsum's disease (tolerable daily intake (TDI) ≤ 10 mg of phytanic acid). The phytol moiety of the PFAE (0.4-5.4 mg/100 g fresh weight) would contribute up to ∼50% to the TDI with the consumption of only one portion of bell pepper fruit pulp.

Method Development for the Determination of Free and Esterified Sterols in Button Mushrooms (Agaricus bisporus).

Ergosterol is the major sterol in button mushrooms (Agaricus bisporus) and can occur as free alcohol or esterified with fatty acids (ergosteryl esters). In this study, gas chromatography with mass spectrometry in the selected ion monitoring mode (GC/MS-SIM) was used to determine ergosterol and ergosteryl esters as well as other sterols and steryl esters in button mushrooms. Different quality control measures were established and sample preparation procedures were compared to prevent the formation of artifacts and the degradation of ergosteryl esters. The final method was then used for the determination of ergosterol (443 ± 44 mg/100 g dry matter (d.m.)) and esterified ergosterol (12 ± 6 mg/100 g d.m.) in button mushroom samples (n = 4). While the free sterol fraction was vastly dominated by ergosterol (∼90% of five sterols in total), the steryl ester fraction was more diversified (nine sterols in total, ergosterol ∼55%) and consisted primarily of linoleic acid esters.