Non-targeted mass spectrometry and feature-based molecular networking for determination of branched fatty acid esters of hydroxy fatty acids in milk.

Branched fatty acid esters of hydroxy fatty acids (FAHFAs) represent trace lipids with significant natural biological functions. While exogenous FAHFAs have been extensively studied, research on FAHFAs in milk remains limited, constraining our grasp of their nutritional roles. This study introduces a non-targeted mass spectrometry approach combined with chemical networking of spectral fragmentation patterns to uncover FAHFAs. Through meticulous sample handling and comparisons of various data acquisition and processing modes, we validate the method's superiority, identifying twice as many FAHFAs compared to alternative techniques. This validated method was then applied to different milk samples, revealing 45 chemical signals associated with known and potential FAHFAs, alongside findings of 66 ceramide/hexosylceramide (Cer/HexCer), 48 phosphatidyl ethanolamine/lyso phosphatidyl ethanolamine (PE/LPE), 21 phosphatidylcholine/lysophosphatidylcholine (PC/LPC), 16 phosphatidylinositol (PI), 7 phosphatidylserine (PS), and 11 sphingomyelin (SM) compounds. This study expands our understanding of the FAHFA family in milk and provides a fast and convenient method for identifying FAHFAs.

Crystallization behavior and compatibility analysis of palm kernel stearin/anhydrous milk fat blends and application.

In this paper, the compatibility, phase behavior, and crystallization properties of the binary blends of palm kernel stearin (PKS) and anhydrous milk fat (AMF) were investigated by analyzing the solid fat content (SFC), thermal properties, polymorphism, and microstructure, with the aim of providing theoretical guidance for the blending of oils. The results showed that the PKS content primarily determined the SFC trend of the binary blends. However, the binary blends demonstrated poor miscibility and eutectic behavior was predominantly observed in the system, particularly at higher temperatures. Only α and ß' forms appeared in this blended system. Simultaneously, the addition of PKS elevated the liquid phase transition temperature of the binary blends, considerably significantly increased their crystallization rate when the addition of PKS was more than 20% and increased the density and size of the fat crystals. Finally, the mixing design optimization method was used to get the most suitable ratio of the binary blends in the refrigerated cream system with PKS:AMF to be 0.914:0.086. The cream prepared with the above binary blends was indeed superior in overrun and firmness and had high stability. PRACTICAL APPLICATION: Some fats with special advantages are often limited in their wide application because of their poor crystallization ability. In this paper, the crystallization ability of fats is improved, and their application scenarios are increased through the combination of fats, so as to provide reference for the production of special fats for food.

The chemical and biological characteristics of fatty acid esters of hydroxyl fatty acids.

With the continuous advancements in detection methods and the exploration of unknown substances, an increasing number of bioactive compounds are being discovered. Fatty acid esters of hydroxyl fatty acids (FAHFAs), a class of endogenous lipids found in 2014, exhibit various physiological activities, such as improving glucose tolerance and insulin sensitivity, stimulating insulin secretion, and demonstrating broad anti-inflammatory effects. Moreover, some FAHFAs are closely linked to intestinal health and can serve as potential biomarkers for gut health. Various FAHFAs have been observed in food, including palmitic acid esters of hydroxy stearic acids (PAHSA), oleic acid esters of hydroxy stearic acids (OAHSA), linoleic acid esters of hydroxy linoleic acid (LAHLA). As a type of lipid regularly consumed in the daily diet, it is highly important to ascertain the types and quantities of FAHFAs present in the diet. This article, based on existing research, provides a review of the analysis methods for FAHFAs, particularly focusing on the separation of chiral isomers. It also summarizes the sources and contents of dietary FAHFAs, emphasizing their bioavailability and impact on the gut. Understanding the beneficial effects of these lipids in the diet can serve as a valuable reference for the development of specific functional foods.

Association of branched-chain fatty acids with metabolic syndrome: a systematic review and meta-analysis of observational studies.

Background: branched-chain fatty acids (BCFAs) have recently emerged as a group of functional fatty acids that are widely distributed in various foodstuffs, including dairy products, ruminant meat products, and fermented foods. Several studies have investigated the differences in the levels of BCFAs among individuals with varying risks of metabolic syndrome (MetS). In this study, we conducted a meta-analysis to explore the relationship between BCFAs and MetS, and to assess the feasibility of BCFAs as potential biomarkers for diagnosing MetS. Methods: in accordance with the PRISMA guidelines, we conducted a systematic literature search on PubMed, Embase, and the Cochrane Library up to March 2023. Both longitudinal and cross-sectional studies were included. The quality of the longitudinal and cross-sectional studies was evaluated using the Newcastle-Ottawa Scale (NOS) and the Agency for Healthcare Research and Quality (AHRQ) criteria, respectively. Heterogeneity detection and sensitivity analysis of the included research literature were carried out using R 4.2.1 software with a random-effects model. Results: Our meta-analysis included 685 participants and revealed a significant negative correlation between the endogenous BCFAs (serum BCFAs and adipose tissue BCFAs) and the risk of developing MetS, with lower BCFA levels found in individuals at a high risk of MetS (WMD: -0.11%, 95% CI: [-0.12, -0.09] %, P < 0.0001). However, there was no difference in fecal BCFAs among different MetS risk groups (SMD: -0.36, 95% CI: [-1.32, 0.61], P = 0.4686). Conclusion: our study provides insights into the relationship between BCFAs and the risk of developing MetS, and lays the groundwork for the development of novel biomarkers for diagnosing MetS in the future.

Monomethyl branched-chain fatty acids-a pearl dropped in the ocean.

As an emerging group of bioactive fatty acids, monomethyl branched-chain fatty acids (mmBCFAs) have sparked the interest of many researchers both domestically and internationally. In addition to documenting the importance of mmBCFAs for growth and development, there is increasing evidence that mmBCFAs are highly correlated with obesity and insulin resistance. According to previous pharmacological investigations, mmBCFAs also exhibit anti-inflammatory effects and anticancer properties. This review summarized the distribution of mmBCFAs, which are widely found in dairy products, ruminants, fish, and fermented foods. Besides, we discuss the biosynthesis pathway in different species and detection methods of mmBCFAs. With the hope to unveil their mechanisms of action, we recapitulated detailed the nutrition and health benefits of mmBCFAs. Furthermore, this study provides a thorough, critical overview of the current state of the art, upcoming difficulties, and trends in mmBCFAs.

Epoxy Triglyceride Enhances Intestinal Permeability via Caspase-1/NLRP3/GSDMD and cGAS-STING Pathways in Dextran Sulfate Sodium-Induced Colitis Mice.

Oxidized triglyceride monomers are the main cytotoxic products of deep-frying oil. However, its impact on the intestinal barrier, the first health guardian, remains unknown. In this study, HPLC-MS/MS analysis revealed that the epoxy group is the main oxidation product, indicating that it may be the main cytotoxic factor. Therefore, 1-9,10-epoxystearic ester, 2,3-dioleic acid (EGT) and glycerol trioleate (GT) were used to reveal the effect of the epoxy group on the intestinal barrier of dextran sulfate sodium-induced colitis. Characteristics analysis showed that EGT could aggravate intestinal damage. The relative mRNA expression analysis suggested that EGT could activate Caspase-1/NLRP3/GSDMD, thereby inducing pyroptosis. The proinflammatory cytokines activated by pyroptosis and the cGAS-STING pathway were released through the pores, thus inducing the disintegration of the tight junction between the intestinal epithelial cells and enhancing intestinal permeability. Metabonomics further confirmed that EGT can change the composition and content of phospholipids on the cell membrane, indicating the morphological changes of the intestinal epithelial cell membrane. In conclusion, this study highlights that EGT induced intestinal dysfunction via Caspase-1/NLRP3/GSDMD and cGAS-STING pathways.

Molecular dynamics revealed the effect of epoxy group on triglyceride digestion.

The digestion behavior of epoxy triglyceride, the main cytotoxic product of deep-frying oil, remains unknown, which may affect its biosafety. In this study, epoxy triglyceride (EGT) and triglyceride (GT) were used to reveal the effect of epoxy group on digestion. Digestibility rate analysis showed that the free fatty acids release rate of EGT was slower. To clarify this phenomenon, binding ability with salt ions in digestive juice and particle size were also been studied. Cluster size analysis indicated that epoxy group increased triglyceride particle size, resulting in smaller contact area between EGT and lipase. Interface behaviors displayed EGT decreased binding ability with salt ions in digestive juice. Spectroscopic analysis showed EGT caused the red shift of lipase peak, indicating that epoxy group changed lipase structure. Molecular dynamics simulation suggested EGT leads to loosen lipase structure. In conclusion, this study highlights that epoxy group could weaken the triglyceride digestion.

Deep-frying oil induces cytotoxicity, inflammation and apoptosis on intestinal epithelial cells.

BACKGROUND: Deep-frying oil has been found to cause inflammatory bowel disease (IBD). However, the molecular mechanism of the effect of deep-frying palm oil on IBD still remains undetermined. RESULTS: In the present study, bioinformatics and cell biology were used to investigate the functions and signal pathway enrichments of differentially expressed genes. The bioinformatics analysis of three original microarray datasets (GSE73661, GSE75214 and GSE126124) in the NCBI-Gene Expression Omnibus database showed 17 down-regulated genes (logFC < 0) and 2 up-regulated genes (logFC > 0) existed in the enteritis tissue. Meanwhile, pathway enrichment and protein-protein interaction network analysis suggested that IBD is relevant to cytotoxicity, inflammation and apoptosis. Furthermore, Caco-2 cells were treated with the main oxidation products of deep-frying oil-total polar compounds (TPC) and its components (polymerized triglyceride, oxidized triglycerides and triglyceride degradation products) isolated from deep-frying oil. The flow cytometry experiment revealed that TPC and its components could induce apoptosis, especially for oxidized triglyceride. A quantitative polymerase chain reaction analysis demonstrated that TPC and its component could induce Caco-2 cell apoptosis through AQP8/CXCL1/TNIP3/IL-1. CONCLUSION: The present study provides fundamental knowledge for understanding the effects of deep-frying oils on the cytotoxic and inflammatory of Caco-2 cells, in addition to clarifying the molecular function mechanism of deep-frying oil in IBD. © 2021 Society of Chemical Industry.

Interactions between Food Hazards and Intestinal Barrier: Impact on Foodborne Diseases.

The intestine is an important digestive organ of the human body, and its barrier is the guardian of the body from the external environment. The impairment of the intestinal barrier is believed to be an important determinant in various foodborne diseases. Food hazards can lead to the occurrence of many foodborne diseases represented by inflammation. Therefore, understanding the mechanisms of the impact of the food hazards on intestinal barriers is essential for promoting human health. This review examined the relationship between food hazards and the intestinal barrier in three aspects: apoptosis, imbalance of gut microbiota, and pro-inflammatory cytokines. The mechanism of dysfunctional gut microbiota caused by food hazards was also discussed. This review discusses the interaction among food hazards, intestinal barrier, and foodborne diseases and, thus, offers a new thought to deal with foodborne disease.

The Triacylglycerol Profile of Oil Bodies and Oil Extracted from Argania spinosa Using the UPLC Along with the Electrospray Ionization Quadrupole-Time-of-Flight Mass Spectrometry (LC-Q-TOF-MS).

The triacylglycerol (TAG) matrix of argan oil (AO) bodies (AOB) along with the TAGs of AO extracted from the same kernels using an organic solvent, were identified and quantified using the ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. Generally, both samples showed a similar TAGs profile but AO found to have three extra TAGs in low amount. In total 23 and 26 different TAGs were identified in AOBs and AO, respectively. The most abundant TAGs were OOL, POO, OOO, and POL in both samples. Furthermore, oleic acid, linoleic acid, and palmitic acid were the major fatty acids in both AOBs and AO. To the best of our knowledge, this is the first research that studied the TAGs matrix of an oil body revealing no major difference between the TAGs profile protected by the AOBs membrane and the oil extracted from the whole seed. PRACTICAL APPLICATION: Seed and kernels oil bodies emulsion tend to be the new source of emulsified oil in food and cosmetic industries. However, before replacing a product with another, we have to make sure that the new alternative can offer better or at least similar benefits. Our results showed that the triacylglycerols (TAGs) matrix and the argan oil (AO) share the same TAGs profile with a relatively close percentage. Therefore, AO bodies can be the perfect pre-emulsified oil for some food products like sauces and creams.

Design, synthesis, biological activities and DFT calculation of novel 1,2,4-triazole Schiff base derivatives.

Series of 1,2,4-triazole Schiff bases (2a-2d, 2f-2h and 3a-3h) have been designed and synthesized. The structure of title compounds was confirmed on the basis of their spectral data and elemental analysis. All the target compounds were screened for their in vitro antifungal activity and antibacterial activity. Two of the tested compounds (2a and 2b) exhibited significant antifungal activity against most fungi, especially compound 2a showed better antifungal activity than triadimefon. Meanwhile, the antibacterial activity assay also indicated compound 2a exhibited excellent antibacterial activities comparable to chloramphenicol. The SAR manifested no substitution at position 5 of the triazole ring caused an increase in activity, and 3-phenoxy phenyl group introduced in 1,2,4-triazole scaffold can enhance the antibacterial activity. The DFT calculation indicated triazole ring, S atom and benzene ring in both of the 2a and 3a make a major contribution to the activity.

Influence of indigenous minor components on fat crystal network of fully hydrogenated palm kernel oil and fully hydrogenated coconut oil.

Purification of triglycerides from fully hydrogenated palm kernel oil (FHPKO) and fully hydrogenated coconut oil (FHCNO) was performed by a chromatographic method. Lipid composition, thermal properties, polymorphism, isothermal crystallization behaviour, nanostructure and microstructure of FHPKO, FHPKO-triacylglycerol (TAG), FHCNO and FHCNO-TAG were evaluated. Removal of minor components had no effect on triglycerides composition. However, the presence of the minor components did increase the slip melting point and promote onset of crystallization. Furthermore, the thickness of the nanoscale crystals increased, and polymorphic transformation from ß' to ß occurred in FHPKO after the removal of minor components, and from α to ß' in FHCNO. Sharp changes in the values of the Avrami constant K and exponent n suggested that the presence of minor components changed the crystal growth mechanism. The PLM results indicated that a coarser crystal structure with lower fractal dimension appeared after the removal of minor components from both FHPKO and FHCNO.

Comparative Analysis of Small-Molecule Diffusivity in Different Fat Crystal Network.

Oil migration and fat recrystallization in fat-structured food materials can result in significant deterioration in food quality. Consequently, it is important to monitor and quantify the diffusivities of the migrants in fat crystal network. The diffusion coefficients of Nile red dye in liquid oils through fully hydrogenated palm kernel oil (FHPKO)/triolein (OOO) and fully hydrogenated soybean oil (FHSO)/triolein (OOO) systems were evaluated by the fluorescence recovery after photobleaching (FRAP) method. The effective diffusion coefficients (Deff) and mobile fraction (Mf) increased with the decrease of solid fat contents (SFC), with the changes of microstructure from more densely to slightly larger packed clusters for both FHPKO/OOO and FHSO/OOO systems. In addition, microstructural parameters of these systems were estimated by the image analysis. The results showed that the diffusion of dye and liquid oil was affected by the microstructure. The higher Deff was associated with lower fractal dimensions, larger crystal thickness, and larger average particle sizes. Finally, higher-permeability coefficients were calculated according to Darcy's Law, and it was significantly correlated to the Deff.

Enzymatically catalyzed synthesis of anti-blooming agent 1,3-dibehenoyl-2-oleoyl glycerol in a solvent-free system: optimization by response surface methodology.

Products rich in 1,3-dibehenoyl-2-oleoyl glycerol (BOB) triglyceride (TAG) were produced by enzymatic interesterification of high oleic acid sunflower oil (HOSO) and behenic acid methyl ester (BME) by 1,3-regiospecific lipase Lipozyme RM IM in a solvent-free system. The impact factors of enzyme load, substrate molar ratio of BME to HOSO (BME/HOSO), reaction time, reaction temperature, and pre-equilibration water activity of the enzyme on BOB content and BME conversions were investigated by single-factor experiments and then optimized using the response surface methodology (RSM). The optimum conditions were as follows: reaction temperature, 72 °C; reaction time, 7.99 h; substrate molar ratio, 2.5:1; enzyme load, 10%; and pre-equilibration water activities of the enzyme, 0.28. The results from the experiments conducted according to the predicted optimal conditions were as follows: the content of BOB was 32.76%, and the conversion of BME was 65.16%. The experimental values agreed with the predicted values, which verified the sufficiency of the quadratic regression models. After purification under the optimal short-range molecular distillation and two-step solvent fractionation, the content of BOB in the target product can reach 77.14%, indicating the great potential for industrial production of the anti-blooming agent.

Model for human milk fat substitute evaluation based on triacylglycerol composition profile.

Being the dominant components in human milk fat (HMF), triacylglycerol (TAG) composition might be the best approximation index to represent the composing characteristics of HMF. In this study, TAG composition of HMF from different lactation stages was analyzed by RP-HPLC-APCI-MS, and the establishment of a model for the precise evaluation of human milk fat substitutes (HMFSs) based on TAG composition was indirectly realized by employment of fatty acid composition and distribution and polyunsaturated fatty acid (PUFA) and TAG compositions. The model was verified by the selected fats and oils with specific chemical compositions, and the results revealed the degrees of similarity of these fats and oils in different evaluation aspects reflected their differences in corresponding chemical composition with HMF. The newly established evaluation model with TAG composition as a comparison base could provide a more accurate method to evaluate HMFSs and might have some inspirations for HMFS production in the future.

Preparation of human milk fat substitutes from palm stearin with arachidonic and docosahexaenoic acid: combination of enzymatic and physical methods.

Human milk fat substitutes (HMFSs) were prepared by a two-step process, namely, Lipozyme RM IM-catalyzed acidolysis of interesterified high-melting palm stearin with fatty acids from rapeseed oil and blending of the enzymatic product with the selected oils on the basis of the calculation model. The optimum conditions for the enzymatic reaction were a mole ratio of palm stearin/fatty acids 1:10, 60 °C, 8% enzyme load (wt % of substrates), 4 h, and 3.5% water content (wt % of enzyme); the enzymatic product contained 39.6% palmitic acid (PA), 83.7% of the fatty acids at sn-2 position were PA (sn-2 PA), and the distribution probability of PA at the sn-2 position among total PA (% sn-2 PA) was 70.5%. With the fatty acid profiles of human milk fat (HMF) as a preferable goal, a physical blending model was established for the second step to guarantee the maximum addition of selected oils. Based on the model prediction, a desirable formula constituted enzymatic product/rapeseed oil/sunflower oil/palm kernel oil/algal oil/microbial oil at a mole ratio of 1:0.28:0.40:0.36:0.015:0.017, and the final product had PA content, sn-2 PA, and %sn-2 PA at 23.5, 43.1, and 61.1%, respectively. The contents of arachidonic and docosahexaenoic acids were 0.4 and 0.3%, respectively. Relying on the total and sn-2 fatty acid compositions of HMF and "deducting score" principle, the score for the similarity between the final product and HMF was scaled as 89.2, indicating the potential as a fat substitute in infant formulas.

Lipase-catalyzed preparation of human milk fat substitutes from palm stearin in a solvent-free system.

Human milk fat substitutes (HMFSs) were synthesized by lipozyme RM IM-catalyzed acidolysis of chemically interesterified palm stearin (mp = 58 °C) with mixed FAs from rapeseed oil, sunflower oil, palm kernel oil, stearic acid, and myristic acid in a solvent-free system. Response surface methodology (RSM) was used to model and optimize the reactions, and the factors chosen were reaction time, temperature, substrate molar ratio, and enzyme load. The optimal conditions generated from the models were as follows: reaction time, 3.4 h; temperature, 57 °C; substrate molar ratio, 14.6 mol/mol; and enzyme load, 10.7 wt % (by the weight of total substrates). Under these conditions, the contents of palmitic acid (PA) and PA at sn-2 position (sn-2 PA) were 29.7 and 62.8%, respectively, and other observed FAs were all within the range of FAs of HMF. The product was evaluated by the cited model, and a high score (85.8) was obtained, which indicated a high degree of similarity of the product to HMF.

Comparative analysis of lipid composition and thermal, polymorphic, and crystallization behaviors of granular crystals formed in beef tallow and palm oil.

Six rectangular block all beef tallow (BT)-based and all palm oil (PO)-based model shortenings prepared on a laboratory scale, respectively denoted BTMS and POMS, were stored under temperature fluctuation cycles of 5-20 °C until granular crystals were observed. The lipid composition and thermal, polymorphic, and isothermal crystallization behaviors of the granular crystals and their surrounding materials separated from BTMS and POMS, respectively, were evaluated. The changes of nanostructure including the aggregation of high-melting triacylglycerols (TAGs) and polymorphic transformation from ß' form of double chain length structures to complicated crystal structures, in which the ß and ß' form crystals of triple and double chain length structures simultaneously coexist, had occurred in granular crystals compared with surrounding materials, whether in BTMS or in POMS. Consequently, a slower crystallization rate appeared in granular crystal parts of both model shortenings noted above, which would yield larger and fewer crystals indicated by the Avrami model analysis that would further aggregate to form large granular crystals.

Characterization of graininess formed in all beef tallow-based shortening.

A batch of all beef tallow (BT)-based model shortening (divided into six rectangular block samples) was stored under temperature fluctuation cycles of 5-20 °C until granular crystals were observed. The lipid composition, thermal properties, and polymorphism of the granular crystals and their surrounding materials were evaluated. Furthermore, the isothermal crystallization behavior of two parts noted above was also examined by pulsed nuclear magnetic resonance (pNMR), rheology, and polarized light microscopy (PLM). The changes of nanostructure including the aggregation of high-melting triacylglycerols (TAGs) and transformation into the most stable ß polymorph occurred in granular crystals compared with surrounding materials. Concomitantly, a slower crystallization rate with a simultaneous increase in crystal growth led to the formation of large crystals and further aggregated to larger granular crystals when the size ultimately exceeded the sensory threshold.

Solvent-free enzymatic preparation of feruloylated monoacylglycerols optimized by response surface methodology.

The ability of immobilized lipase B from Candida antarctica (Novozym 435) to catalyze the direct esterification of glyceryl ferulate (FG) and oleic acid for feruloylated monoacylglycerols (FMAG) preparation in a solvent-free system was investigated. Enzyme screening and the effect of glycerol on the initial reaction rate of esterification were also investigated. Response surface methodology (RSM) was used to optimize the effects of the reaction temperature (55-65 degrees C), the enzyme load (8-14%; relative to the weight of total substrates), oleic acid/(FG + glycerol) (6:1-9:1; w/w), and the reaction time (1-2 h) on the conversion of FG and yield of FMAG. Validation of the RSM model was verified by the good agreement between the experimental and the predicted values of FG conversion and FMAG yield. The optimum preparation conditions were as follows: temperature, 60 degrees C; enzyme load, 8.2%; substrate ratio, 8.65:1 (oleic acid/(FG + glycerol), w/w); and reaction time, 1.8 h. Under these conditions, the conversion of FG and yield of FMAG are 96.7 +/- 1.0% and 87.6 +/- 1.2%, respectively.