Differentiation of sea buckthorn syrups processed by high pressure, pulsed electric fields, ohmic heating, and thermal pasteurization based on quality evaluation and chemical fingerprinting.

Introduction: Impact of processing on product characteristics, sustainability, traceability, authenticity, and public health along the food chain becomes more and more important not only to the producer but also to the customer and the trust of a consumer toward a brand. In recent years, the number of juices and smoothies containing so called super foods or fruits, which have been "gently pasteurized," has increased significantly. However, the term "gentle pasteurization" related to the application of emerging preservation technologies such as pulsed electric fields (PEF), high pressure processing (HPP) or ohmic heating (OH) is not clearly defined. Methods: Therefore, the presented study investigated the influence of PEF, HPP, OH, and thermal treatment on quality characteristics and microbial safety of sea buckthorn syrup. Syrups from two different varieties were investigated under the following conditions HPP (600 MPa 4-8 min), OH (83°C and 90°C), PEF (29.5 kV/cm, 6 µs, 100 Hz), and thermal (88°C, hot filling). Analyses to test the influence on quality parameters like ascorbic acid (AA), flavonoids, carotenoids, tocopherols, antioxidant activity; metabolomical/chemical profiling (fingerprinting) via U-HPLC-HRMS/MS (here especially flavonoids and fatty acids); sensory evaluation, as well as microbial stability including storage, were conducted. Results and discussion: Independent from the treatment, the samples were stable over 8 weeks of storage at 4°C. The influence on the nutrient content [Ascorbic acid (AA), total antioxidant activity (TAA), total phenolic compounds (TPC), tocopherols (Vit E)] was similar for all tested technologies. Employing statistical evaluation Principal Component Analysis (PCA) a clear clustering based on the processing technologies was observed. Flavonoids as well as fatty acids were significantly impacted by the type of used preservation technology. This was obvious during the storage time of PEF and HPP syrups, where enzyme activity was still active. The color as well as taste of the syrups were found to be more fresh-like for the HPP treated samples.

Effect of temperature on the compositions of ladderane lipids in globally surveyed anammox populations.

The adaptation of bacteria involved in anaerobic ammonium oxidation (anammox) to low temperatures will enable more efficient removal of nitrogen from sewage across seasons. At lower temperatures, bacteria typically tune the synthesis of their membrane lipids to promote membrane fluidity. However, such adaptation of anammox bacteria lipids, including unique ladderane phospholipids and especially shorter ladderanes with absent phosphatidyl headgroup, is yet to be described in detail. We investigated the membrane lipids composition (UPLC-HRMS/MS) and dominant anammox populations (16S rRNA gene amplicon sequencing, Fluorescence in situ hybridization) in 14 anammox enrichments cultivated at 10-37 °C. "Candidatus Brocadia" appeared to be the dominant organism in all but two laboratory enrichments of "Ca. Scalindua" and "Ca. Kuenenia". At lower temperatures, the membranes of all anammox populations were composed of shorter [5]-ladderane ester (reduced chain length demonstrated by decreased fraction of C20/(C18 + C20)). This confirmed the previous preliminary evidence on the prominent role of this ladderane fatty acid in low-temperature adaptation. "Ca. Scalindua" and "Ca. Kuenenia" had distinct profile of ladderane lipids compared to "Ca. Brocadia" biomasses with potential implications for adaptability to low temperatures. "Ca. Brocadia" membranes contained a much lower amount of C18 [5]-ladderane esters than reported in the literature for "Ca. Scalindua" at similar temperature and measured here, suggesting that this could be one of the reasons for the dominance of "Ca. Scalindua" in cold marine environments. Furthermore, we propose additional and yet unreported mechanisms for low-temperature adaptation of anammox bacteria, one of which involves ladderanes with absent phosphatidyl headgroup. In sum, we deepen the understanding of cold anammox physiology by providing for the first time a consistent comparison of anammox-based communities across multiple environments.

The Effect of Mycotoxins and Silymarin on Liver Lipidome of Mice with Non-Alcoholic Fatty Liver Disease.

Milk thistle-based dietary supplements have become increasingly popular. The extract from milk thistle (Silybum marianum) is often used for the treatment of liver diseases because of the presence of its active component, silymarin. However, the co-occurrence of toxic mycotoxins in these preparations is quite frequent as well. The objective of this study was to investigate the changes in composition of liver lipidome and other clinical characteristics of experimental mice fed by a high-fat methionine-choline deficient diet inducing non-alcoholic fatty liver disease. The mice were exposed to (i) silymarin, (ii) mycotoxins (trichothecenes, enniatins, beauvericin, and altertoxins) and (iii) both silymarin and mycotoxins, and results were compared to the controls. The liver tissue extracts were analyzed by ultra-high performance liquid chromatography coupled with high-resolution tandem mass spectrometry. Using tools of univariate and multivariate statistical analysis, we were able to identify 48 lipid species from the classes of diacylglycerols, triacylglycerols, free fatty acids, fatty acid esters of hydroxy fatty acids and phospholipids clearly reflecting the dysregulation of lipid metabolism upon exposure to mycotoxin and/or silymarin.

Metabolomic fingerprinting as a tool for authentication of grapevine (Vitis vinifera L.) biomass used in food production.

Use of 'green biomass' of the grapevine is gradually extending into the food industry. The aim of our study was to demonstrate the potential of metabolomic fingerprinting for characterization of grapevine leaves and canes. Our method comprises successive aqueous-methanolic extractions, followed by U-HPLC-HRMS/MS. For data processing, PCA and (O)PLS-DA methods were utilized, and mathematical models were validated. We showed that from all factors investigated, harvesting season explained most of the variability between samples, followed by locality combined with farming system. The identified statistically significant metabolites for harvesting season models mostly represented the groups of fatty acids, fatty phenols, (lyso)phospholipids, flavonoids and organic acids. For models of localities with different farming systems, majority of identified metabolites significant for organic farming belonged to groups of fatty acids and their derivatives, terpenoids, sterols, and fat soluble vitamins, whereas for conventional farming, the only identified significant metabolites were the pesticide residues.

A Non-Targeted High-Resolution Mass Spectrometry Study for Extra Virgin Olive Oil Adulteration with Soft Refined Oils: Preliminary Findings from Two Different Laboratories.

This work presents a non-targeted high-resolution mass spectrometry inter-laboratory study for the detection of new chemical markers responsible of soft refined oils addition to extra virgin olive oils. Refined oils (soft deodorized and soft deacidified) were prepared on a laboratory scale starting from low-quality olive oils and analyzed together with a set of pure extra virgin olive oil (EVOO) samples and with mixtures of adulterated and pure EVOO at different percentages. The same analytical workflow was applied in two different laboratories equipped with two types of instrumentation (Q-Orbitrap and Q-TOF); a group of discriminant molecules was selected, and a tentative identification of compounds was also proposed. In summary, 12 molecules were identified as markers of this specific adulteration, and seven of them were selected as discriminative in both the laboratories, with a similar trend throughout the samples (i.e., propylene glycol 1 stearate). The results obtained in the two laboratories are comparable, concretely demonstrating the inter-laboratory repeatability of non-targeted studies. As a confirmation, the same markers were detected also in "in-house" mixtures and in suspect commercial deodorized mixtures, reinforcing the robustness of the results obtained and proving that, thanks to these molecules, mixtures containing at least 40% of adulterated oils can be detected.

Cranberries versus lingonberries: A challenging authentication of similar Vaccinium fruit.

Due to unique phytochemicals contained, Vaccinum berries are known to have a number of positive health effects. In this context, lingonberries (Vaccinium vitis-idaea) are considered to be the most effective, thus finding many uses. Recently, fraud suspicion on lingonberries-based products has been reported, partial or even total replacement by less valued cranberries (Vaccinium macrocarpon) was found. In this study, metabolomic fingerprinting employing instrumental platform consisting of U-HPLC-HRMS/MS was investigated for discrimination between the two Vaccinum berries species. Methanolic extracts of 33 authentic samples from two harvest years were analyzed and chemometric evaluation was performed to identify significant marker compounds, their stability during drying process was assessed, too. The characteristic markers most contributing to berries classification were representatives of polyphenols and phospholipids. Peonidin 3-O-arabinoside and myricetin 3-O-glucoside, not occurring in lingonberries, enabled to discover the presence of cranberries in prepared admixtures down to 1% (w/w).

Analysis of phosphodiesterase type 5 inhibitors as possible adulterants of botanical-based dietary supplements: extensive survey of preparations available at the Czech market.

Popularity of natural-based preparations supporting the sexual potency significantly increased in recent years, which also led to the increase of illegal use of phosphodiesterase type 5 inhibitor (PDE-5) in sexual performance enhancement products. In this study, a rapid U-HPLC‒HRMS/MS method has been developed to simultaneously determine 59 PDE-5 inhibitors and their analogues. Within the development of sensitive method for analysis of 59 PDE-5 inhibitors and their analogues, both sample preparation procedure, as well as separation / detection conditions have been optimized. Extraction efficiency of particular extraction solvents, influence of different mobile phase additives on target analytes separation, as well as impact of various settings of mass analyzer on sensitivity of detection were examined. Data were collected in the 'full MS/data dependent MS/MS' acquisition mode (full MS-dd-MS/MS). Before the U-HPLC‒HRMS/MS method was used for analysis of real samples, proper validation had been conducted. The precision of the method expressed as the relative standard deviation (RSD) was ≤4.2% and ≤5.2% at spiking concentrations 5 µg/g and 0.25 µg/g, respectively. The limits of quantification were in the range 0.25 - 0.05 µg/g and the recovery ranged between 71 and 90%. The optimized method was successfully applied for analysis of 64 real samples, and 10 of them were proved to contain both registered or unregistered synthetic PDE-5 inhibitors. Additionally, the acquired U-HPLC‒HRMS/MS fingerprints were demonstrated to serve as an efficient tool for revealing of other type of possible fraud in products labeling. Retrospective mining of markers of herbs declared on dietary supplements packaging allowed to assess the trueness / untruth in the declaration of medical herbs composition.

A novel approach based on untargeted lipidomics reveals differences in the lipid pattern among durum and common wheat.

In the present work the possibility of using an untargeted metabolomic strategy to discriminate between common and durum wheat lipidome for an authenticity purpose was explored. A first study was conducted by analyzing 52 samples from two durum and common wheat varieties. Afterwards, an extended and independent sample set (173 samples and five varieties) was used as a confirmatory study to verify the stability and consistency of the models obtained. Putatively identified markers were evaluated applying ROC curves resulting in individual marker AUC >90% both in preliminary and confirmatory study. In addition, digalactosyl diglyceride (DGDG) 36:4 was shown to be an effective marker differentiating between authentic durum wheat and its adulterated admixture down to 3% adulteration level, which is the maximum contamination level allowed by Italian legislation. The results demonstrated that untargeted lipidomics, in conjunction with chemometric tools has a significant potential for screening and detection of wheat fraud.

Impact of vacuum frying on quality of potato crisps and frying oil.

This research was focused on a critical assessment of vacuum frying as a technology enabling minimization of acrylamide formation in potato crisps and reducing undesirable chemical changes that occur in frying oil at high temperatures. The potato slices were fried in rapeseed oil under vacuum at 125°C and atmospheric pressure at 165°C. The experiments were performed on two potato varieties, Saturna and Impala. Vacuum frying reduced the formation of acrylamide by 98% and also other Maillard reaction products, specifically alkylpyrazines. Concurrently a lower extent of oxidative changes was observed in the frying oil, while 3-MCPD esters decreased fairly quickly during conventional frying. Sensory characteristics of the vacuum and conventionally fried potato crisps were evaluated by a 23-member panel. The majority of panellists preferred the flavour of 'conventional crisps', while only a few of them appreciated potato-like fresh flavour of 'vacuum crisps' and classified this product as 'tasty'.

Untargeted metabolomics reveals links between Tiger nut (Cyperus esculentus L.) and its geographical origin by metabolome changes associated with membrane lipids.

Tiger nuts and tiger nut milk are well-known Valencian products, and step-by-step these tubers and the tuber-based beverage are becoming more and more relevant products in international markets. However, the increasing demand and success of Valencian tiger nuts did not allow protected designation of origin (PDO) tuber to supply the domestic and international markets. Therefore, the verification of the geographical origin is highly required. In this research, the main objective was to combine an advance analytical method and chemometrics tools in order to decipher the geographical origin of 45 tiger nut samples from (i) 'Xufa de València' PDO and (ii) African samples. The analytical method, based on solid-liquid extraction followed by ultra-high performance liquid chromatography coupled to high resolution tandem mass spectrometry (UHPLC-HRMS) metabolomics approach, highlighted sensitivity and wide linear dynamic range in order to simultaneously analyse polar and non-polar metabolites. After data processing, a pronounced sample clustering according to the geographical origin was clearly observed using unsupervised models, and supervised models revealed that tiger nuts lipidome was associated with the geographical origin. As a result, African samples highlighted an overexpression of phospholipids, such as phosphatidylethanolamine 34:1, and triacylgricerols crosslinked to environmental stress and alteration of membrane lipid compositions.

Untargeted metabolomics of fresh and heat treatment Tiger nut (Cyperus esculentus L.) milks reveals further insight into food quality and nutrition.

Tiger nut (Cyperus esculentus L.) is a crop traditionally grown in Valencia Region (Spain) and other temperate and tropical regions in the world, where its tubers are commonly consumed as tiger nut milk (horchata). Because of their nutritive potential and original taste, these products are beginning to spread internationally and, as consequence, analytical procedures to assess nutritional profiles, quality control issues are acquiring increasing relevance. The main objective of this study was to use an advance analytical method and chemometrics tools to determine if the ultra-high temperature (UHT) treatment necessary to extend the shelf life of tiger nut milk would affect the profile of nutrients when compared to fresh product. A cold solvent extraction followed by liquid chromatography coupled with high resolution mass spectrometry (UHPLC-HRMS) was used. Datasets obtained from UHT and fresh tiger nut milk data were analyzed through an untargeted metabolomics approach to compare chemical patterns, highlighting differences in citric acid esters of mono- diglycerides (CITREM) and monoacylglycerol (MAG) used as emulsifiers of UHT products, and a remarkably higher abundance of biotin, phosphatidic acid (PA) and L-arginine in fresh products. These results showed that untargeted metabolomics through high resolution tandem mass spectrometry allowed fine differences between food products to be found, therefore, the nutrient lost caused by UHT treatment was clearly discerned.