Position Of Bond Research Articles

Varying the position of phospholipid acyl chain unsaturation modulates hopanoid and sterol ordering

The cell membrane must balance mechanical stability with fluidity to function as both a barrier and an organizational platform. Key to this balance is the ordering of hydrocarbon chains and the packing of lipids. Many eukaryotes synthesize sterols, which are uniquely capable of modulating the lipid order to decouple membrane stability from fluidity. Ancient sterol analogs known as hopanoids are found in many bacteria and proposed as ancestral ordering lipids. The juxtaposition of sterols and hopanoids in extant organisms prompts us to ask why both pathways persist, especially in light of their convergent ability to order lipids. In this work, simulations, monolayer experiments, and cellular assays show that hopanoids and sterols order unsaturated phospholipids differently based on the position of double bonds in the phospholipid acyl chain. We find that cholesterol and diplopterol’s methyl group distributions lead to distinct effects on unsaturated lipids. In Mesoplasma florum, diplopterol’s constrained ordering capacity reduces membrane resistance to osmotic stress, unlike cholesterol. These findings suggest that cholesterol’s broader lipid-ordering ability may have facilitated the exploration of a more diverse lipidomic landscape in eukaryotic membranes.

Intramolecular Disulfide Bridges in Voltage-Dependent Anion Channel 2 (VDAC2) Protein from Rattus norvegicus Revealed by High-Resolution Mass Spectrometry.

Voltage-Dependent Anion Channel isoforms (VDAC1, VDAC2, and VDAC3) are relevant components of the outer mitochondrial membrane (OMM) and play a crucial role in regulation of metabolism and in survival pathways. As major players in the regulation of cellular metabolism and apoptosis, VDACs can be considered at the crossroads between two broad families of pathologies, namely, cancer and neurodegeneration, the former being associated with elevated glycolytic rate and suppression of apoptosis in cancer cells, the latter characterized by mitochondrial dysfunction and increased cell death. Recently, we reported the characterization of the oxidation pattern of methionine and cysteines in rat and human VDACs showing that each cysteine in these proteins is present with a preferred oxidation state, ranging from the reduced to the trioxidized form, and such an oxidation state is remarkably conserved between rat and human VDACs. However, the presence and localization of disulfide bonds in VDACs, a key point for their structural characterization, have so far remained undetermined. Herein we have investigated by nanoUHPLC/High-Resolution nanoESI-MS/MS the position of intramolecular disulfide bonds in rat VDAC2 (rVDAC2), a protein that contains 11 cysteines. To this purpose, extraction, purification, and enzymatic digestions were carried out at slightly acidic or neutral pH in order to minimize disulfide bond interchange. The presence of six disulfide bridges was unequivocally determined, including a disulfide bridge linking the two adjacent cysteines 4 and 5, a disulfide bridge linking cysteines 9 and 14, and the alternative disulfide bridges between cysteines 48, 77, and 104. A disulfide bond, which is very resistant to reduction, between cysteines 134 and 139 was also detected. In addition to the previous findings, these results significantly extend the characterization of the oxidation state of cysteines in rVDAC2 and show that it is highly complex and presents unusual features. Data are available via ProteomeXchange with the identifier PXD044041.

The effect of gD-derived peptides on T cell immune response mediated by BTLA-HVEM protein complex in melanoma patients.

The effector function of T cells is regulated via immune checkpoints, activating or inhibiting the immune response. The BTLA-HVEM complex, the inhibitory immune checkpoint, may act as one of the tumor immune escape mechanisms. Therefore, interfering with the binding of these proteins can prove beneficial in cancer treatment. Our study focused on peptides interacting with HVEM at the same place as BTLA, thus disrupting the BTLA-HVEM interaction. These peptides' structure and amino acid sequences are based on the gD protein, the ligand of HVEM. Here, we investigated their immunomodulatory potential in melanoma patients. Flow cytometry analyses of activation, proliferation, and apoptosis of T cells from patients were performed. Additionally, we evaluated changes within the T cell memory compartment. The most promising compound - Pep(2), increased the percentages of activated T cells and promoted their proliferation. Additionally, this peptide affected the proliferation rate and apoptosis of melanoma cell line in co-culture with T cells. We conclude that the examined peptide may act as a booster for the immune system. Moreover, the adjuvant and activating properties of the gD-derived peptide could be used in a combinatory therapy with currently used ICI-based treatment. Our studies also demonstrate that even slight differences in the amino acid sequence of peptides and any changes in the position of the disulfide bond can strongly affect the immunomodulatory properties of compounds.

Structural elucidation of complex polyesters polyols from bio-lubricant using off-line liquid chromatography x supercritical fluid chromatography coupled with Orbitrap mass spectrometry

Synthetic complex esters and polyol esters are incorporated as partially bio-based and biodegradable alternatives to petroleum base oils in lubricant formulations, to provide specific properties or performance and to help reducing their carbon footprint in certain cases. A sample can contain over 400 molecules of high chemical similarity including numerous isomers. To resolve such complexity, a separation technique with large peak capacity coupled to high-resolution mass spectrometry (HRMS) is essential. In this study, comprehensive off-line LCxSFC hyphenated with an Orbitrap analyzer was used for the structural elucidation of a synthetic bio-lubricant composed of a polyol reacted with fatty acids of varying length or with repetitive units of polyesters of ricinoleic acid. Retention in the LC first dimension was mostly due to the degree of oligomerization of ricinoleic acid within the polyester and to the chain length of the fatty acid. The SFC second dimension highlighted the esterification degree of the polyalcohol and the number and positions of fatty acids double bonds. The combination of both dimensions permitted the separation of isomers. The coupling of SFC with Orbitrap analyzer allowed an accurate assignment of molecular formulas. Finally, the fragmentation in the ionization source confirmed the attributed structures. By introducing a clear distribution of the chemical structures in the retention space, LCxSFC-HRMS provided a powerful analytical method for the comprehensive molecular characterization of the complex polyester polyols sample.

Anti-inflammatory lindenane sesquiterpenoid dimers from the roots of Chloranthus holostegius var. trichoneurus.

Two new lindenane-type sesquiterpenoid dimers, chlotrichenes C and D (1 and 2) together with five known lindenane-type sesquiterpenoid dimers (3-7) were isolated from the roots of Chloranthus holostegius var. trichoneurus, a famous natural medicine named as "Sikuaiwa" for subduing swellings and relieving pain. The structures including absolute configuration were elucidated by their 1D and 2D NMR, HRESIMS, and ECD data. Compounds 1 and 2 were classical [4 + 2] lindenane-type sesquiterpenoid dimers that differed from known analogs in oxidation profile, side chain profile, and double bond position. The new isolates and compound 3 exhibited significant inhibitory activity on IL-1β production (IC50: 1-15μM) in LPS-induced THP-1 cells and other compounds exhibited inhibitory activity on NO production in LPS-induced RAW 264.7 cells (IC50: 24-33μM).

Search friction, liquidity risk, and bond misallocation

Search friction is a key driver of changes in corporate bond yield spreads over time. In the cross-section, the liquidity risk stemming from search friction is significantly priced, and is strongly correlated with the misallocation of bond positions among different traders. I propose a novel measure of bond-specific misallocation, which is the negative covariance between traders’ private valuations and their inventory positions for each bond. I find that bonds with higher levels of misallocation are associated with lower absolute levels of liquidity risk from search friction. I develop a search-and-matching model to explain this correlation.

Evolutionary Conservation in Protein-Protein Interactions and Structures of the Elongator Sub-Complex ELP456 from Arabidopsis and Yeast.

The Elongator complex plays a pivotal role in the wobble uridine modification of the tRNA anticodon. Comprising two sets of six distinct subunits, namely, Elongator proteins (ELP1-ELP6) and associated proteins, the holo-Elongator complex demonstrates remarkable functional and structural conservation across eukaryotes. However, the precise details of the evolutionary conservation of the holo-Elongator complex and its individual sub-complexes (i.e., ELP123; ELP456) in plants remain limited. In this study, we conducted an in vivo analysis of protein-protein interactions among Arabidopsis ELP4, ELP5, and ELP6 proteins. Additionally, we predicted their structural configurations and performed a comparative analysis with the structure of the yeast Elp456 sub-complex. Protein-protein interaction analysis revealed that AtELP4 interacts with AtELP6 but not directly with AtELP5. Furthermore, we found that the Arabidopsis Elongator-associated protein, Deformed Roots and Leaves 1 (DRL1), did not directly bind to AtELP proteins. The structural comparison of the ELP456 sub-complex between Arabidopsis and yeast demonstrated high similarity, encompassing the RecA-ATPase fold and the positions of hydrogen bonds, despite their relatively low sequence homology. Our findings suggest that Arabidopsis ELP4, ELP5, and ELP6 proteins form a heterotrimer, with ELP6 serving as a bridge, indicating high structural conservation between the ELP456 sub-complexes from Arabidopsis and yeast.

Disulfide Bonds of Thyroid Peroxidase Are Critical Elements for Subcellular Localization, Proteasome-Dependent Degradation, and Enzyme Activity.

Background: Congenital hypothyroidism (CH) is caused by mutations in cysteine residues, including Cys655 and Cys825 that form disulfide bonds in thyroid peroxidase (TPO). It is highly likely that these disulfide bonds could play an important role in TPO activity. However, to date, no study has comprehensively analyzed cysteine mutations that form disulfide bonds in TPO. In this study, we induced mutations in cysteine residues involved in disulfide bonds formation and analyzed their effect on subcellular localization, degradation, and enzyme activities to evaluate the importance of disulfide bonds in TPO activity. Methods: Vector plasmid TPO mutants, C655F and C825R, known to occur in CH, were transfected into HEK293 cells. TPO activity and protein expression levels were measured by the Amplex red assay and Western blotting. The same procedure was performed in the presence of MG132 proteasome inhibitor. Subcellular localization was determined using immunocytochemistry and flow cytometry. The locations of all disulfide bonds within TPO were predicted using in silico analysis. All TPO mutations associated with disulfide bonds were induced. TPO activity and protein expression levels were also measured in all TPO mutants associated with disulfide bonds using the Amplex red assay and Western blotting. Results: C655F and C825R showed significantly decreased activity and protein expression compared with the wild type (WT) (p < 0.05). In the presence of the MG132 proteasome inhibitor, the protein expression level of TPO increased to a level comparable with that of the WT without increases in its activity. The degree of subcellular distribution of TPO to the cell surface in the mutants was lower compared with the WT TPO. Twenty-four cysteine residues were involved in the formation of 12 disulfide bonds in TPO. All TPO mutants harboring an amino acid substitution in each cysteine showed significantly reduced TPO activity and protein expression levels. Furthermore, the differences in TPO activity depended on the position of the disulfide bond. Conclusions: All 12 disulfide bonds play an important role in the activity of TPO. Furthermore, the mutations lead to misfolding, degradation, and membrane insertion.

Using Subcritical Water to Obtain Polyphenol-Rich Extracts with Antimicrobial Properties.

The use of green extraction methods that meet the criteria of sustainable and environmentally friendly technologies has been increasing in recent decades due to their many benefits. In this respect, extracts obtained using subcritical water are also gaining increased attention because of their potential antioxidant and antimicrobial properties. Their antimicrobial activity is mainly due to the presence of various polyphenolic compounds. Although the exact mechanism of the antibacterial action of polyphenolic compounds has not yet been fully investigated and described, polyphenols are known to affect the bacterial cell at several cellular levels; among other things, they cause changes and ruptures in the cell membranes of the bacterial cell, affect the inactivation of bacterial enzymes and damage bacterial DNA. The difference in the strength of the antimicrobial activity of the extracts is most likely a result of differences in their lipophilicity and in the number and position of hydroxyl groups and double bonds in the chemical structure of polyphenols. By changing the extraction conditions, especially the temperature, during subcritical water extraction, we affect the solubility of the compounds we want to extract. In general, as the temperature increases, the solubility of polyphenolic compounds also increases, and the reduction of the surface tension of subcritical water at higher temperatures also enables faster dissolution of polyphenolic compounds. Different bacterial strains have different sensitivity to different extracts. However, extracts obtained with subcritical water extraction demonstrate strong antimicrobial activity compared to extracts obtained with conventional methods.

Concomitant investigation of crustacean amphipods lipidome and metabolome during the molting cycle by Zeno SWATH data-independent acquisition coupled with electron activated dissociation and machine learning

BackgroundDIA (Data-Independent Acquisition) is a powerful technique in Liquid Chromatography coupled with high-resolution tandem Mass Spectrometry (LC-MS/MS) initially developed for proteomics studies and recently emerging in metabolomics and lipidomics. It provides a comprehensive and unbiased coverage of molecules with improved reproducibility and quantitative accuracy compared to Data-Dependent Acquisition (DDA). Combined with the Zeno trap and Electron-Activated Dissociation (EAD), DIA enhances data quality and structural elucidation compared to conventional fragmentation under CID. These tools were applied to study the lipidome and metabolome of the freshwater amphipod Gammarus fossarum, successfully discriminating stages and highlighting significant biological features. Despite being underused, DIA, along with the Zeno trap and EAD, holds great potential for advancing research in the omics field. ResultsDIA combined with the Zeno trap enhances detection reproducibility compared to conventional DDA, improving fragmentation spectra quality and putative identifications. LC coupled with Zeno-SWATH-DIA methods were used to characterize molecular changes in reproductive cycle of female gammarids. Multivariate data analysis including Principal Component Analysis and Partial Least Square Discriminant Analysis successfully identified significant features. EAD fragmentation helped to identify unknown features and to confirm their molecular structure using fragmentation spectra database annotation or machine learning. EAD database matching accurately annotated five glycerophospholipids, including the position of double bonds on fatty acid chain moieties. SIRIUS database predicted structures of unknown features based on experimental fragmentation spectra to compensate for database incompleteness. SignificanceReproducible detection of features and confident identification of putative compounds are pivotal stages within analytical pipelines. The DIA approach combined with Zeno pulsing enhances detection sensitivity and targeted fragmentation with EAD in positive polarity provides orthogonal fragmentation information. In our study, Zeno-DIA and EAD thereby facilitated a comprehensive and insightful exploration of pertinent biological molecules associated with the reproductive cycle of gammarids. The developed methodology holds great promises for identifying informative biomarkers on the health status of an environmental sentinel species.

Monitoring Changes to Alkenone Biosynthesis in Commercial Tisochrysis lutea Microalgae.

Alkenones are unique lipids produced by certain species of microalgae, well-known for use in paleoclimatology, and more recently pursued to advance sustainability across multiple industries. Beginning in 2018, the biosynthesis of alkenones by commercially grown Tisochrysis lutea (T-Iso) microalgae from one of the world's most established producers, Necton S.A., changed dramatically from structures containing 37 and 38 carbons, to unusual shorter-chain C35 and C36 diunsaturated alkenones (C35:2 and C36:2 alkenones). While the exact reasons for this change remain unknown, analysis of alkenones isolated from T-Iso grown in 2021 and 2023 revealed that this change has persisted. The structure of these rare shorter-chain alkenones, including double bond position, produced by Necton T-Iso remained the same over the last five years, which was determined using a new and optimized cross-metathesis derivatization approach with analysis by comprehensive two-dimensional gas chromatography and NMR. However, noticeable differences in the alkenone profiles among the different batches were observed. Combined with fatty acid compositional analysis, the data suggest a connection between these lipid classes (e.g., increased DHA corresponds to lower amounts of shorter-chain alkenones) and the ability to manipulate their biosynthesis in T-Iso with changes to cultivation conditions.

Oleic Acid Could Act as a Channel Blocker in the Inhibition of nAChR: Insights from Molecular Dynamics Simulations.

The activation of the muscular nicotinic acetylcholine receptor (nAChR) produces the opening of the channel, with the consequent increase in the permeability of cations, triggering an excitatory signal. Free fatty acids (FFA) are known to modulate the activity of the receptor as noncompetitive antagonists, acting at the membrane-AChR interface. We present molecular dynamics simulations of a model of nAChR in a desensitized closed state embedded in a lipid bilayer in which distinct membrane phospholipids were replaced by two different monounsaturated FFA that differ in the position of a double bond. This allowed us to detect and describe that the cis-18:1ω-9 FFA were located at the interface between the transmembrane segments of α2 and γ subunits diffused into the channel lumen with the consequent potential ability to block the channel to the passage of ions.

Insignoic acids A – E, unusual α, β-unsaturated keto fatty acids isolated from the exocarp of Australian rainforest tree Endiandra insignis (Lauraceae)

Anti-inflammatory bioassay-guided compound isolation from the exocarp of the Australian rainforest tree Endiandra insignis (family Lauraceae) has led to the discovery and structural elucidation of unusual α, β-unsaturated twenty-four carbon fatty acids and their positional isomers, insignoic acids A – E (1a – 5c). The stereochemistry and position of the double bond within the aliphatic chain were independently determined via NMR spectroscopy and Ozone-Induced Dissociation (OzID) Mass Spectrometry, respectively. Compounds (1a – 5c) displayed good to moderate anti-inflammatory activity in the range of 8–84 μM. The low therapeutic index observed when assessing the cell viability in the RAW macrophage cell lines, prompted us to investigate the anticancer potential of these unusual fatty acids. The anti-cancer activity was assessed in A-431 carinoma cell lines and MM649 melanoma cell lines. Insignoic acid C (3a-f) exhibited the highest level of potency with an IC50 value of 5–7 μM against both the cell lines. The insignoic acids are the first of their kind known for incorporating an alpha-beta unsaturated system flanked next to a keto group with an additional level of oxygenation at C-6 in a 24‑carbon fatty acid backbone.

Quantifying Fluorescence Lifetime Responsiveness of Environment-Sensitive Probes for Membrane Fluidity Measurements.

The structural diversity of different lipid species within the membrane defines its biophysical properties such as membrane fluidity, phase transition, curvature, charge distribution, and tension. Environment-sensitive probes, which change their spectral properties in response to their surrounding milieu, have greatly contributed to our understanding of such biophysical properties. To realize the full potential of these probes and avoid misinterpretation of their spectral responses, a detailed investigation of their fluorescence characteristics in different environments is necessary. Here, we examined the fluorescence lifetime of two newly developed membrane order probes, NR12S and NR12A, in response to alterations in their environments such as the degree of lipid saturation, cholesterol content, double bond position and configuration, and phospholipid headgroup. As a comparison, we investigated the lifetime sensitivity of the membrane tension probe Flipper in these environments. Applying fluorescence lifetime imaging microscopy (FLIM) in both model membranes and biological membranes, all probes distinguished membrane phases by lifetime but exhibited different lifetime sensitivities to varying membrane biophysical properties (e.g., cholesterol). While the lifetime of Flipper is particularly sensitive to the membrane cholesterol content, the NR12S and NR12A lifetimes are moderately sensitive to both the cholesterol content and lipid acyl chains. Moreover, all of the probes exhibit longer lifetimes at longer emission wavelengths in membranes of any complexity. This emission wavelength dependency results in varying lifetime resolutions at different spectral regions, which are highly relevant for FLIM data acquisition. Our data provide valuable insights on how to perform FLIM with these probes and highlight both their potential and limitations.

Experimental study on bond-slip behavior of vertical connecting rebar in the horizontal joints of double- superimposed shear wall

The horizontal joint of double-superimposed shear wall is the important part affecting the seismic performance of the wall. The horizontal joint mainly relies on the synergistic action between the vertical connecting rebar and the cast-in-place concrete to transfer the horizontal and vertical loadings and coordinate the deformation. Therefore, it is necessary to study the bond-slip behavior between the vertical connecting rebar and the cast-in-place concrete. In this paper, the effects of the cast-in-place layer thickness, the concrete strength, the anchorage length, and the diameter of rebar on the interface bond properties of vertical connecting rebar were analyzed through pullout tests. Based on the experimental results, the calculation formulas for bond strength and peak slip were proposed. According to the distribution of bond stress along the anchorage length, the bond-slip constitutive model optimized by the bond position function was established to describe the bond-slip behavior between the vertical connecting rebar and the cast-in-place concrete.

Paternò-Büchi Reaction Mass Spectrometry Enables Positional Assignment of Polymethylene-Interrupted Double Bonds in Food-Derived Lipids.

Fatty acids (FAs) containing polymethylene-interrupted (PMI) double bonds are a component of human foods; however, they present a significant analytical challenge for de novo identification. Covalent adduct chemical ionization and ozone-induced dissociation mass spectrometry (MS) methods enable unambiguous assignment of PMI-FA double bond positions, however, no method has been reported with electrospray ionization (ESI) platform using off-the-shelf systems. In the current work, we studied the Paternò-Büchi (PB) fragmentation patterns of PMI-FA and triacylglycerol (TG) by analyzing several known food sources. PB-MS/MS and MS3 enabled complete double bond location assignments, including the isolated double bond in PMI-FA and triacylglycerols. Sea urchin ("uni"), oyster, pine nut, and ginkgo nut were characterized for their signature PMI-FA, 20:2(5Z,11Z), 22:2(7Z, 15Z), 18:3(5Z,9Z,12Z), and 20:3(5Z,11Z,14Z), respectively. Quantitative analyses of the relative abundance of these PMI-FA led to results similar to reference methods. 18:3(5Z,9Z,12Z) was enriched at the sn-1/sn-3 position in pine nut major TG.

Interactions of small molecule inhibitors with secreted phospholipase A2: A review of the structural data

AbstractThe secreted phospholipase A2 (sPLA2) family of enzymes hydrolyzes glycerophospholipid substrates at the sn‐2 position of the acyl ester bond. Besides their roles in the degradation of phospholipids, these enzymes also play an important role in the immune response by promoting inflammation. The products of sPLA2‐catalysis (lysophospholipids and free fatty acids such as arachidonic acid) as well as secondary metabolites produced by the arachidonic acid cascade are mediators of inflammation and subsequent tissue injury. Based on their physiological functions, the sPLA2 enzymes are pharmacological targets for the treatment of diseases with an inflammatory component such as atherosclerosis and rheumatoid arthritis. Interestingly, sPLA2s may also have roles in various types of cancer such as malignant prostate cancer. While a number of sPLA2 inhibitors have been described, compounds which have been co‐crystallized with human sPLA2 enzymes are of much importance as such three‐dimensional structures provide valuable information regarding the structural requirements of good potency inhibitors as well as the molecular interactions between the enzyme and inhibitor. This review selected four structures of human sPLA2s complexed with inhibitors from the Protein Data Bank to illustrate the important molecular interactions of inhibitor compounds with the enzymes and to highlight the structural features important for interaction with the sPLA2 active site. In addition, the complex of the sPLA2 inhibitor, varespladib, with a Lys49‐PLA2‐like snake venom toxin is also discussed. Knowledge of the molecular interactions acquired from such structures would greatly aid the discovery of small molecule inhibitors of sPLA2s for the treatment of snakebite envenoming. The three‐dimensional structures presented in this review have been published by various research groups and have been deposited in the Protein Data Bank.

Systematic quantum chemical tailoring to the molecular structure, optical and nonlinear optical properties of N/B dopped C60 buckminsterfullerene

Buckminsterfullerene is hard to study experimentally and computationally owing to its unique reactivity and larger molecular size, respectively. The whole C60 fullerene molecule consists of two types of chemical bonds i.e. hexagon-hexagon bonds and hexagon-pentagon bonds. A series of heterofullerenes (C58hhBN-C58hpBN) was designed by substitution in C60 fullerene with nitrogen (N) and boron (B) at the hexagon-hexagon bond and hexagon-pentagon bond positions. Quantum chemical methods were employed to examine the structural and optoelectronic characteristics of the designed compounds as compared to parent C60. The average polarizability measurements shown that the linear response of the C58hpBB compound (αiso = 116.7 × 10−24 esu, αaniso = 10.56 × 10−24 esu) is dominant. Among all designed compounds, a change in hexagon-pentagon bonds is seen as more effective for tuning the NLO response properties. For instance, C58hpBB and C58hpNN showed the larger <γ> response of 164.5 × 10−36 and 172.5 × 10−36 esu indicating greater potential for NLO materials. A number of techniques such as natural bond orbitals (NBOs), molecular electrostatic potential (MEPs), frontier molecular orbitals (FMOs) and absorption spectra were used to investigate the optoelectronic properties. The maximum absorption of molecules are found in between 569 and 800 nm. According to FMO analysis, C58hpBB and C58hpNN have smaller energy gaps (0.97 eV and 1.47 eV) and better intramolecular charge transfer. The current study evokes the aspect of hexagon-hexagon bond and hexagon-pentagon bond modification for efficient dopped buckminsterfullerene.

The number and position of unsaturated bonds in aliphatic aldehydes affect meat flavorings system: Insights on initial Maillard reaction stage and meat flavor formation from thiazolidine derivatives

Nonanal, (E)-2-nonenal, (E,E)-2,4-nonadienal, and (E,Z)-2,6-nonadienal were used to study the effect of number and position of the unsaturated bond in aliphatic aldehydes on meat flavorings. Cysteine-Amadori and thiazolidine derivatives were synthesized, identified by UPLC-TOF/MS and NMR, and quantitatively by UPLC-MS/MS. The polyunsaturated aldehydes exhibited higher inhibition than monounsaturated aldehydes, and monounsaturated aldehydes exhibited higher inhibition than saturated aldehydes, mainly manifested by the inhibition of the cysteine-Amadori formation and acceleration of the thiazolidine derivatives formation. The effect of unsaturated bonds position in aliphatic aldehydes on the initial Maillard reaction stage was similar. The cysteine played an important role in catalyzing the reaction of aliphatic aldehydes. A total of 109 volatile compounds derived by heating prepared thiazolidine derivatives degradation were detected by GC-MS. Formation pathways of volatile compounds were proposed by retro-aldol, oxidation, etc. Particularly, a route to form thiazole by the decarboxylation reaction of thiazolidine derivatives which derivatives from formaldehyde reacting with cysteine was proposed.

Resolving phytosterols in microalgae using offline two-dimensional reversed phase liquid chromatography-supercritical fluid chromatography coupled with quadrupole time-of-flight mass spectrometry.

Sterols are unsaponifiable lipids resulting from plant metabolism that exhibit interesting bioactive properties. Microalgae are a major source of specific phytosterols, most of which are still not fully characterized. The similarity in sterol structures and the existence of positional isomers make the separation of phytosterols challenging. A method was developed based on an offline two-dimensional (2D) system, reversed-phase liquid chromatography (RPLC)-supercritical fluid chromatography (SFC)/quadrupole time-of-flight (Q-ToF) mass spectrometry, for the identification of sterols in microalgae. Subsequent positive-mode MS/MS was used to confirm the identified phytosterols. The 2D chromatogram exhibited a pattern related to the positions of the double bonds, which were confirmed by standard injection, enabling structural elucidation. The analysis of the unsaponifiable fraction of two algae, namely Scenedesmus obliquus, a freshwater microalgae, and Padina pavonica, a marine macroalgae, highlighted the ability of the method to distinguish a large number of sterol isomers.