Untargeted Lipidomics Method for the Discrimination of Five Crab Species by Ultra-High-Performance Liquid Chromatography High-Resolution Mass Spectrometry Combined with Chemometrics.

In this study, ultra-high-performance liquid chromatography high-resolution accurate mass-mass spectrometry (UHPLC-HRAM/MS) was applied to characterize the lipid profiles of five crab species. A total of 203 lipid molecular species in muscle tissue and 176 in edible viscera were quantified. The results indicate that Cancer pagurus contained high levels of lipids with a docosahexaenoic acid (DHA) and eicosapntemacnioc acid (EPA) structure in the muscle tissue and edible viscera. A partial least squares discriminant analysis (PLS-DA) showed that PE 16:0/22:6, PE P-18:0/20:5, PA 16:0/22:6 and PC 16:0/16:1 could be used as potential biomarkers to discriminate the five kinds of crabs. In addition, some lipids, such as PE 18:0/20:5, PC 16:0/16:1, PE P-18:0/22:6 and SM 12:1;2O/20:0, could be used as characteristic molecules to distinguish between Cancer magister and Cancer pagurus, which are similar in appearance. This study provides a new perspective on discriminating crab species from MS-based lipidomics.

Untargeted Metabolomic Analysis Combined with Chemometrics Revealed the Effects of Different Cooking Methods on Lentinus edodes.

Cooking methods affect the compositions of Lentinus edodes metabolites. Nevertheless, little information is available on the specific impact of different cooking methods on Lentinus edodes via metabolomic analysis. This study determined the influence of boiling, steaming, air-frying, and roasting on the metabolomic profiles of Lentinus edodes based on UHPLC-Q-Exactive Orbitrap MS/MS in combination with chemometrics. A total of 990 metabolites were detected and classified into 11 super-classes. Subsequently, the metabolites of the four cooking methods were distinguished using multivariate statistical analysis. The results showed that boiling caused a massive loss of metabolites while roasting and air-frying led to an evident upregulation. The upregulation of metabolites in the steaming groups was not as significant as in roasting and air-frying. This study provided reference data for a comprehensive understanding of the metabolites associated with domestic cooking methods and valuable guidance for the development of Lentinus edodes and its products in the future.

Discrimination of chrysanthemum varieties using lipidomics based on UHPLC-HR-AM/MS/MS.

BACKGROUND: Chrysanthemum is one of the most important and popular ornamentals over the world. Chrysanthemum drink is a type of traditional healthy drink like Chinese tea. Owing to the differences in the chemical compositions, different chrysanthemum varieties have different medicinal effects on human health. Thus, the identification of different chrysanthemum varieties is very important and necessary. This study aims to distinguish seven chrysanthemum varieties that are widely used in China. First, total lipids were obtained from chrysanthemums. After that, lipid profiles were characterized using ultra-high-performance liquid chromatography hyphenated with a Q Exactive™ high resolution-accurate-mass mass spectrometer. RESULTS: A total of 163 lipid molecular species from 17 types of lipid classes in seven varieties of chrysanthemums were determined. Principal component analysis indicated that three lipid molecules, lysophosphatidylethanolamine(18:2) (LPE(18:2)), LPE(16:0), and phosphatidic acid(18:2/18:3) (variable importance in projection >3, P < 0.001), can be used as potential biomarkers to distinguish seven chrysanthemum varieties. Hierarchical cluster analysis showed that the lipid molecular profiles of 'Gongju' were most similar to 'Jinzijianju', followed by 'Huaibaiju', 'Boju', 'Hangbaiju', 'Chuju', and 'Fubaiju'. CONCLUSION: This comprehensive analysis provided a new method to identify chrysanthemum varieties through the perspective of lipidomics combined with chemometrics. © 2022 Society of Chemical Industry.

Cytotoxic and Pro-Apoptotic Effects of Leaves Extract of Antiaris africana Engler (Moraceae).

Antiaris africana Engler leaves have been used in Senegalese folk medicine to treat breast cancer. The present study aimed to investigate the anticancer potential of Antiaris africana Engler leaves using several human cancer cell lines. The leaves of Antiaris africana Engler were extracted in parallel with water or 70% ethanol and each extract divided into three parts by successive liquid-liquid extraction with ethyl acetate and butanol. The phytochemical components of the active extract were investigated using ultra-performance liquid chromatography-diode array detector-quadrupole time-of-flight tandem mass spectrometry (UPLC-DAD-QTOF-MS/MS). The cytotoxic and cytostatic effects of each extract, as well as their fractions, were evaluated in vitro via flow and image cytometry on different human cancer phenotypes, such as breast (MCF-7), pancreas (AsPC-1), colon (SW-620) and acute monocytic leukemia (THP-1). Both hydro-alcoholic and aqueous extracts induced strong apoptosis in MCF-7 cells. The water fraction of the hydro-alcoholic extract was found to be the most active, suppressing the cell growth of MCF-7 in a dose-dependent manner. The half maximum effective concentration (EC50) of this fraction was 64.6 ± 13.7 µg/mL for MCF-7, with equivalent values for all tested phenotypes. In parallel, the apoptotic induction by this fraction resulted in a EC50 of 63.5 ± 1.8 µg/mL for MCF-7, with again equivalent values for all other cellular tested phenotypes. Analysis of this fraction by UPLC-DAD-QTOF-MS/MS led to the identification of hydroxycinnamates as major components, one rutin isomer, and three cardiac glycosides previously isolated from seeds and bark of Antiaris africana Engler and described as cytotoxic in human cancer models. These results provide supportive data for the use of Antiaris africana Engler leaves in Senegal.

Deep eutectic solvent-based headspace single-drop microextraction for the quantification of terpenes in spices.

Deep eutectic solvents (DESs) were investigated as extracting solvent for headspace single-drop microextraction (HS-SDME). The extraction efficiency of 10 DESs mainly composed of tetrabutylammonium bromide (N4444Br) and long-chain alcohols was evaluated for the extraction of terpenes from six spices (cinnamon, cumin, fennel, clove, thyme, and nutmeg). The DES composed of N4444Br and dodecanol at a molar ratio of 1:2 showed the highest extraction efficiency and was selected to conduct the extractions of terpenes in the rest of the study. HS-SDME was optimized by design of experiments. Only two parameters from the four studied showed a significant influence on the efficiency of the method: the extraction time and the extraction temperature. The optimal extraction conditions were determined by response surface methodology. All extracts were analyzed by gas chromatography coupled to mass spectrometry (GC-MS). More than 40 terpenes were extracted and identified in nutmeg, the richest extract in terpenes in this study. Quantitative analysis based on 29 standards was conducted for each extract. Good linearity was obtained for all standards (R2 > 0.99) in the interval of 1 to 500 µg/g. Limits of quantification ranged from 0.47 µg/g (borneol) to 86.40 µg/g (α-farnesene) with more than half of the values under 2 µg/g. HS-SDME is simple, rapid, and cheap compared with conventional extraction methods. The use of DESs makes this extraction method "greener" and it was shown that DESs can be suitable solvents for the extraction of bioactive compounds from plants.

Beneficial effects of cherry consumption as a dietary intervention for metabolic, hepatic and vascular complications in type 2 diabetic rats.

BACKGROUND: Oxidative stress (OS) plays an important role in type 2 diabetes (T2D) pathogenesis and its complications. New therapies target natural antioxidants as an alternative and/or supplemental strategy to prevent and control them. Our previous chemical and biological studies highlighted the important antioxidant activities of cherries, among other fruits and vegetables, thus we aimed to determine in vivo effects of 2-month long cherry consumption using a high-fat/high-fructose (HFHF) model of diabetic-rats (Lozano et al. in Nutr Metab 13:15, 2016). METHODS: After 2 months of HFHF, male Wistar rats were divided into: HFHF and HFHF enriched in cherry (nutritional approach) or standard diet ND (lifestyle measures) and ND plus cherry during 2 months. Metabolic, lipidic, oxidative parameters were quantified. Tissues (liver, pancreas and vessels) OS were assessed and hepatic (steatosis, fibrosis, inflammation) and vascular (endothelial dysfunction) complications were characterized. RESULTS: T2D was induced after 2 months of HFHF diet, characterized by systemic hyperglycaemia, hyperinsulinemia, glucose intolerance, dyslipidaemia, hyperleptinemia, and oxidative stress associated with endothelial dysfunction and hepatic complications. Cherry consumption for 2 months, in addition to lifestyle measures, in T2D-rats decreased and normalized the systemic disturbances, including oxidative stress complications. Moreover, in the vessel, cherry consumption decreased oxidative stress and increased endothelial nitric oxide (NO) synthase levels, thus increasing NO bioavailability, ensuring vascular homeostasis. In the liver, cherry consumption decreased oxidative stress by inhibiting NADPH oxidase subunit p22phox expression, nuclear factor erythroid-2 related factor 2 (Nrf2) degradation and the formation of reactive oxygen species. It inhibited the activation of sterol regulatory element-binding proteins (1c and 2) and carbohydrate-responsive element-binding protein, and thus decreased steatosis as observed in T2D rats. This led to the improvement of metabolic profiles, together with endothelial and hepatic function improvements. CONCLUSION: Cherry consumption normalized vascular function and controlled hepatic complications, thus reduced the risk of diabetic metabolic disorders. These results demonstrate that a nutritional intervention with a focus on OS could prevent and/or delay the onset of vascular and hepatic complications related to T2D.

On-Line Screening, Isolation and Identification of Antioxidant Compounds of Helianthemum ruficomum.

Many Helianthemum species (Cistaceae) are recognized for their various medicinal virtues. Helianthemum ruficomum is an endemic species to the septentrional Sahara on which no report is available so far. The purpose of this work was to investigate the chemical composition and the radical scavenging capacity of this species and its isolated components. Collected from Mougheul (south-west of Algeria), the aerial parts were macerated with 80% EtOH/H2O, after evaporation, the remaining extract was diluted with H2O and extracted with petroleum ether, chloroform, ethyl acetate and n-butanol. EtOAc and n-BuOH extracts were evaluated for their free radical scavenging capacity by on-line HPLC-ABTS•+ assay. The obtained data which were confirmed by TEAC and ORAC assays, allowed guiding the fractionation of these extracts by CC, TLC and reverse phase HPLC. Among the components, 14 were isolated and identified by spectroscopic analyses: protocatechuic acid (1), trans-tiliroside (2), cis-tiliroside (3), astragalin (4), picein (7), vanillic acid 4-O-ß-d-glucopyranoside (8), lavandoside (9), 4-hydroxybenzoic acid 4-O-ß-d-glucopyranoside (10), nicotiflorin (11), rutin (12), vicenin-2 (13), narcissin (14) and stigmasterol (5) and ß-sitosterol (6) as a mixture (71% and 29%, respectively). Compounds 5, 7, 8, 9, 10 and 14 were new for the genus Helianthemum. The antioxidant power of all the isolated compounds was also evaluated by HPLC-ABTS•+, TEAC and ORAC assays. The results clearly indicated high antioxidant potential of the extracts and tested compounds of this species especially, compounds 1, 4, 8, 9, 10 and 12.

Dietary walnut oil modulates liver steatosis in the obese Zucker rat.

PURPOSE: Non-alcoholic fatty liver disease (NAFLD) is the hepatic manifestation of the metabolic syndrome. We aimed to clarify the impact of dietary walnut oil versus animal fat on hepatic steatosis, representing the initial step of multistage pathogenesis of NAFLD, in Zucker obese rats. METHODS: Zucker lean ad libitum (a.l.), Zucker obese a.l. or Zucker obese pair fed (p.f.) to the lean received isocaloric diets containing 8% walnut oil (W8), W14 or 14% lard (L14) (n = 10/group). Body weight, clinical serology, liver weight, lipid content and fatty acid composition and hepatic lipid metabolism-related transcripts were evaluated. RESULTS: Compared to lean, Zucker obese a.l. and p.f. showed hepatic triacylglyceride (TAG) accumulation. In Zucker obese p.f., W14 compared to W8 and L14 reduced liver lipids, TAG as well as hepatic omega-6 (n-6)/n-3 ratio and SCD activity index [(C18:0 + C18:1)/C18:0 ratio] paralleled by decreased lipoprotein lipase mRNA in obese p.f. and elevated microsomal triglyceride transfer protein mRNA in lean and obese. Further, W14 elevated the fasting blood TAG and reduced cholesterol levels in obese. CONCLUSIONS: In our model, consumption of W14 inhibited hepatic lipid accumulation along with modulated hepatic gene expression implicated in hepatic fatty acid influx or lipoprotein assembly. These results provide first indication that dietary lipids from walnut oil are modulators of hepatic steatosis as the initial step of progressive NAFLD pathogenesis.

In silico ADMET and anti-inflammatory profiles of the stigmast-4-en-3-one and isolation of isovanillin as the antioxidant principle of Imperata cylindrica (L.) P. Beauv.

Imperata cylindrica (L.) P. Beauv. is an invasive species widely used in treatment of several diseases associated with pain and inflammation in different countries including Madagascar. This work aims to report the isolation of the antioxidant, analgesic and anti-nflammatory compounds from the methanol extract of I. cylindrica. The bio-guided method was used to isolate its bioactive compounds by combining chromatographic methods, writhing test in mice and antioxidant assays. Stigmast-4-en-3-one was isolated as one among the compounds responsible for the analgesic and anti-inflammatory properties and isovanillin as one among the antioxidant compounds from the extract. Stigmast-4-en-3-one showed a good oral pharmacokinetic profile and good binding affinities with some pro-inflammatory targets. It did not show any mutagenic effect, nor a carcinogenic one and had a low risk to be a cardiotoxic agent. All of our results provide scientific justification for its traditional medicinal use in the management of pain and inflammatory related diseases.

Identification of volatiles from oxidised phosphatidylcholine molecular species using headspace solid-phase microextraction (HS-SPME) and gas chromatography-mass spectrometry (GC-MS).

Headspace solid-phase microextraction (HS-SPME) followed by gas chromatography-mass spectrometry analysis (GC-MS) was used to investigate the volatile compounds from oxidised phosphatidylcholine molecular species. 1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine (SOPC) and 1-stearoyl-2-linoleoyl-sn-glycero-3-phosphocholine (SLPC) were chosen as models. The influence of several parameters on the efficiency of volatile oxidised compounds (VOCs) microextraction, such as type of fibre, extraction duration and temperature were studied. The best results were obtained with a polydimethylsiloxane/divinylbenzene (PDMS/DVB) fibre used at 50 °C during 25 min. The effect of oxidation temperature on the yield of VOCs from SOPC and SLPC was investigated. Oxidative kinetics of SOPC and SLPC were investigated by measuring both the production of VOCs and the degradation of starting materials. More than 30 VOCs were detected by means of the reference mass spectra of the National Institute of Standards and Technology mass spectral library, and most of them were further confirmed by comparing their mass spectra and retention time with those obtained from authentic reference compounds under the same analytical conditions. Moreover, the origins of VOCs from oxidised PLs were studied by comparing those obtained from their corresponding triacylglycerides under the same experimental conditions. The main VOCs identified from oxidised SOPC were (E)-2-decenal, nonanal and octanal and from oxidised SLPC were (E)-2-heptenal, (E)-2-octenal and (E, E)-2,4-decadienal. The proposed method was applied to a real food sample, soy lecithin.

Protein-Based High Internal Phase Pickering Emulsions: A Review of Their Fabrication, Composition and Future Perspectives in the Food Industry.

Protein-based high internal phase Pickering emulsions (HIPEs) are emulsions using protein particles as a stabilizer in which the volume fraction of the dispersed phase exceeds 74%. Stabilizers are irreversibly adsorbed at the interface of the oil phase and water phase to maintain the droplet structure. Protein-based HIPEs have shown great potential for a variety of fields, including foods, due to the wide range of materials, simple preparation, and good biocompatibility. This review introduces the preparation routes of protein-based HIPEs and summarizes and classifies the preparation methods of protein stabilizers according to their formation mechanism. Further outlined are the types and properties of protein stabilizers used in the present studies, the composition of the oil phase, the encapsulating substances, and the properties of the constituted protein-based HIPEs. Finally, future development of protein-based HIPEs was explored, such as the development of protein-based stabilizers, the improvement of emulsification technology, and the quality control of stabilizers and protein-based HIPEs.

Characterization and discrimination of volatile organic compounds and lipid profiles of truffles under different treatments by UHPLC-QE Orbitrap/MS/MS and P&T-GC-MS.

The lipid profiles of the truffles with different treatments were determined by ultra-high-performance liquid chromatography-Quadrupole-Exactive Orbitrap mass spectrometry (UHPLC-QE Orbitrap/MS/MS) and the volatile organic compounds (VOCs) were identified by purge-and-trap-gas chromatography-mass spectrometry (P&T-GC-MS). A total of 37 lipid molecular species and 28 VOCs were tentatively identified. Lysophophatidylcholine (LPC), triacylglycerol (TG) and sphingomyelin (SM) in heat-drying truffles, phosphatidic acid (PA) in freeze-drying and fresh truffles might be the key lipids that bound VOCs. Furthermore, the correlation between lipids and VOCs were analyzed by 19 differential lipids and 7 VOCs. The findings indicated that TG 18:2/18:2/18:2 and Cardiolipin (CL) 16:0/16:0/18:2/18:2 might be the key lipid molecule species for the formation of 2-methoxyphenol. The study helps to understand the effect of different treatments on the lipid profiles and provides the mechanistic insights to the relationship between the lipids and VOCs of truffles.

Construction of whey protein gels prepared by three methods to stabilize high internal phase Pickering emulsions loaded with CoQ10 under different pH.

The aim of this study was to investigate the effect of whey protein gel particles (WPGPs) prepared by heat-induced method, enzyme cross-linking method and calcium ion cross-linking method on the structural properties and intrinsic linkage of their stable high internal phase Pickering emulsions (HIPPEs) under different pH conditions. The effects of different pH and preparation methods on the internal interaction forces, particle size, ζ-potential, wettability and secondary structure of gels was investigated. The results indicated that the construction of HIPPEs system was successfully constructed at pH 3, 5 or 7. The WPGPs stabilized HIPPEs can maintain stable state at 4 °C for 28 days. Coenzyme Q10 (CoQ10) loaded with HIPPEs increased the bioavailability from 13.2% to 79.4%, which was demonstrated in in vitro digestion experiments.

Revealing the dynamic changes of lipids in coffee beans during roasting based on UHPLC-QE-HR-AM/MS/MS.

Coffee is popular worldwide and its consumption is increasing in recent years. Although mass spectrometry-based lipidomics approaches have been prevalent, their application in studies related to detailed information and dynamic changes in lipid composition during coffee bean roasting is still limited. The aim of this study was to investigate the dynamic changes in coffee bean lipids during the roasting process. The lipid classes and lipid molecular species in coffee beans were characterized by lipidomic analysis combined with chemometrics. A total of 12 lipid classes and 105 lipid molecular species were identified and quantified. Triacylglycerols (TAG) was the most abundant lipid class in both green beans and roasted beans. The content of phosphatidylethanolamine (PE) and lysophosphatidylethanolamine (LPE) in green beans was obviously higher than that in roasted beans. Other phospholipids, such as phosphatidylinositol (PI), lysophosphatidylinositol (LPI), phosphatidylcholine (PC), lysophophatidylcholine (LPC) and phosphatidic acid (PA), showed a tendency to increase at the beginning of roasting, then decreased gradually. Several differential lipid molecule species, for instance, PE (16:0_18:2), PC (18:2_18:2) were significantly down-regulated, and PI (18:1_18:2) was significantly up-regulated. This study provided a scientific basis for the change of coffee bean lipids during the roasting process.

Unraveling the potential of breath and sweat VOC capture devices for human disease detection: a systematic-like review of canine olfaction and GC-MS analysis.

The development of disease screening methods using biomedical detection dogs relies on the collection and analysis of body odors, particularly volatile organic compounds (VOCs) present in body fluids. To capture and analyze odors produced by the human body, numerous protocols and materials are used in forensics or medical studies. This paper provides an overview of sampling devices used to collect VOCs from sweat and exhaled air, for medical diagnostic purposes using canine olfaction and/or Gas Chromatography-Mass spectrometry (GC-MS). Canine olfaction and GC-MS are regarded as complementary tools, holding immense promise for detecting cancers and infectious diseases. However, existing literature lacks guidelines for selecting materials suitable for both canine olfaction and GC-MS. Hence, this review aims to address this gap and pave the way for efficient body odor sampling materials. The first section of the paper describes the materials utilized in training sniffing dogs, while the second section delves into the details of sampling devices and extraction techniques employed for exhaled air and sweat analysis using GC-MS. Finally, the paper proposes the development of an ideal sampling device tailored for detection purposes in the field of odorology. By bridging the knowledge gap, this study seeks to advance disease detection methodologies, harnessing the unique abilities of both dogs and GC-MS analysis in biomedical research.

Combining Untargeted Lipidomics Analysis and Chemometrics to Identify the Edible and Poisonous Mushrooms (Pleurotus cornucopiae vs Omphalotus japonicus).

The global phenomenon of eating poisonous mushrooms by mistake occurs every year. Untargeted lipidomics analysis combined with chemometrics was used to identify mushroom varieties. Two kinds of mushrooms with similar appearance, namely, Pleurotus cornucopiae (P. cornucopiae) and Omphalotus japonicus (O. japonicus) were selected as models, where O. japonicus was a poisonous mushroom and P. cornucopiae was an edible mushroom. First, the lipid extraction efficiency of eight solvents was compared. The methyl tert-butyl ether/methanol (2:1, v/v) had higher lipid extraction efficiency of extracting mushroom lipids than other solvents, in terms of the lipid coverage, response intensity, and solvent safety. Afterward, the comprehensive lipidomics analysis of the two mushrooms was conducted. A total of 21 lipid classes and 267 molecular species were identified in O. japonicus, whereas 22 lipid classes and 266 molecular species in P. cornucopiae. The principal component analysis demonstrated that 37 characteristic metabolites, including TAG 18:1_18:2_18:0;1O, TAG 18:1_18:1_18:2, TAG 16:2_18:2_18:2, etc., could be used to distinguish the two mushrooms. These differential lipids were able to identify P. cornucopiae blended with 5% (w/w) O. japonicus. This study explored a novel method for identifying poisonous mushrooms from edible mushrooms and provided a reference for food safety of consumers.

Investigation of biomarkers of bile tolerance in Lactobacillus casei using comparative proteomics.

The identification of cell determinants involved in probiotic features is a challenge in current probiotic research. In this work, markers of bile tolerance in Lactobacillus casei were investigated using comparative proteomics. Six L. casei strains were classified on the basis of their ability to grow in the presence of bile salts in vitro. Constitutive differences between whole cell proteomes of the most tolerant strain (L. casei Rosell-215), the most sensitive one (L. casei ATCC 334), and a moderately tolerant strain (L. casei DN-114 001) were investigated. The ascertained subproteome was further studied for the six strains in both standard and bile stressing conditions. Focus was on proteins whose expression levels were correlated with observed levels of bile tolerance in vitro, particularly those previously reported to be involved in the bile tolerance process of lactobacilli. Analysis revealed that 12 proteins involved in membrane modification (NagA, NagB, and RmlC), cell protection and detoxification (ClpL and OpuA), as well as central metabolism (Eno, GndA, Pgm, Pta, Pyk, Rp1l, and ThRS) were likely to be key determinants of bile tolerance in L. casei and may serve as potential biomarkers for phenotyping or screening purposes. The approach used enabled the correlation of expression levels of particular proteins with a specific probiotic trait.

Determination of phosphatidylethanolamine molecular species in various food matrices by liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS2).

A liquid chromatographic-electrospray ionization-tandem mass spectrometric (LC-ESI-MS(2)) method has been developed for determination of the molecular species of phosphatidylethanolamine (PE) in four food matrices (soy, egg yolk, ox liver, and krill oil). The extraction and purification method consisted of a pressurized liquid extraction procedure for total lipid (TL) extraction, purification of phospholipids (PLs) by adsorption on a silica gel column, and separation of PL classes by semi-preparative normal-phase HPLC. Separation and identification of PE molecular species were performed by reversed-phase HPLC coupled with electrospray ionization tandem mass spectrometry (ESI-MS(2)). Methanol containing 5 mmol L(-1) ammonium formate was used as the mobile phase. A variety of PE molecular species were detected in the four food matrices. (C16:0-C18:2)PE, (C18:2-C18:2)PE, and (C16:0-C18:1)PE were the major PE molecular species in soy. Egg yolk PE contained (C16:0-C18:1)PE, (C18:0-C18:1)PE, (C18:0-C18:2)PE, and (C16:0-C18:2)PE as the major molecular species. Ox liver PE was rich in the species (C18:0-C18:1)PE, (C18:0-C20:4)PE, and (C18:0-C18:2)PE. Finally, krill oil which was particularly rich in (C16:0(alkyl)-C22:6(acyl))plasmanylethanolamine (PakE), (C16:0-C22:6)PE, and (C16:0-C20:5)PE, seemed to be an interesting potential source for supplementation of food with eicosapentaenoic acid and docosahexaenoic acid.

Improvement in determination of isothiocyanates using high-temperature reversed-phase HPLC.

The largely adopted reversed-phase HPLC analysis of the molecular species of isothiocyanates (ITCs) was performed and showed losses during the chromatographic run with eight ITCs. These losses, which obviously impact the accuracy of quantitative determinations, were due to precipitation in the chromatographic system. At 22°C, they ranged from 5.4% for sulforaphane (SFN) to 11.0% for benzyl-ITC when ITCs were injected at 80 µg mL(-1) , but they were up to three times higher at 1 mg mL(-1) reaching 31.9% for benzyl-ITC. The water solubility of the ITCs was a key determinant of the extent of the measured loss. When the column was heated at 60°C, losses in injected ITCs were reduced, in comparison with 22, 40, and 50°C, by two to ten times depending on the ITC considered. A reversed-phase HPLC method based on column heating was suggested and its quantitative performance was determined. It was then applied to the separation of methylene chloride extracts of various cruciferous vegetables. Ally-ITC, SFN, and iberin in cabbage; SFN and iberin in cauliflower; and allyl-ITC and phenylethyl-ITC in horseradish could be identified and quantified. The obtained results cast doubt on quantitative determinations of ITCs that are carried out at room temperature using reversed-phase HPLC.

SPE for the simultaneous determination of various isothiocyanates.

Several SPE sorbents were investigated for the extraction of a group of chemically diverse isothiocyanates (ITCs). They included bonded silica, carbon-based, and polymer-based sorbents with various functional groups. Results showed large differences in the ability of these sorbents to simultaneously extract ITCs from standard solutions. Recovery rates were on average the highest with divinylbenzene (DVB) based polymeric sorbents, especially with a DVB/N-vinylpyrrolidone copolymer that had recovery rates ranging between 86.7 and 95.6%. These sorbents achieved the most balanced extraction efficiency between aliphatic and aromatic, polar, and nonpolar ITCs. With graphitized carbon, C(18)-bonded silica, and amide-containing sorbent, recovery levels were higher for the two least polar aromatic ITCs (benzyl ITC and phenylethyl ITC), whereas for the polar aliphatic ITCs levels were the lowest. The least retained one, was methyl ITC that is the most polar with recoveries between 0 and 31.5%. The presence of amide groups, especially in a polyamide sorbent, appeared to be particularly unsuitable for the extraction of aliphatic ITCs. A copolymer made up of DVB and N-vinylpyrrolidone was therefore shown to be the most suited for the extraction of both aliphatic and aromatic ITCs.