Liquid Chromatography-electrospray Mass Spectrometry Research Articles

Assessment of variable agroclimatic impact on dissipation kinetics of ready-mix fungicide formulation in green chili for harmonization of food safety

Multi-locational supervised field trials were conducted with ready-mix formulation of two fungicides, tricyclazole (18%) + mancozeb (62%) in green chili (Capsicum annuum) to assess the impacts of variable weather conditions on the residual profile for more realistic food safety evaluation. Liquid chromatography-Electrospray Ionization-Mass Spectrometry [LC-ESI(+ve)-MS/MS] and Ultraviolet Visible (UV-Vis) spectrophotometer have been employed for tricyclazole and mancozeb determination respectively. Tricyclazole was extracted in acetone: water (9:1) with dispersive solid phase extraction (d-SPE) cleaned up whereas mancozeb was digested in concentrated HCl and SnCl2. The developed method was successfully validated as per SANTE/12682/2019 guidelines. On green chilli, 225 + 775 g a.iha−1(active ingredient per hectare) (1250 g formulation-ha−1) and 450 + 1550 g a.iha−1 (active ingredient per hectare) (2500 g formulation-ha−1) of a combined formulation of tricyclazole (18%) and mancozeb (62%), WP, were employed as the recommended dose (T1) and double the recommended dose (T2), respectively. The initial depositions of tricyclazole were varied from 0.47 to 0.74 and 1.30–2.33 mg kg−1 and for mancozeb ranged from 0.93 to 1.31 and 2.68–4.13 mg kg−1 at the respective doses. The respective half-life (days) values of tricyclazole and mancozeb were 1.65–2.41 and 1.20–1.66 irrespective of doses and locations. The maximum residue limit (MRL) of 0.05 mg kg−1 for tricyclazole in green chili has been proposed for Food Safety and Standard Authority of India (FASSAI), India with recommended pre-harvest intervals (PHI) of 12 days. The third location (Anand Agricultural University, Anand, Gujarat) with the least rainfall and humidity has recorded the highest initial deposition, persistence, half-life, and PHI. The ready-mix fungicide has also increased fresh yield up to 22–34%. All these data may provide valuable information on the safe use and promotion of fungicides mixture.

Optimization of a method to detect levothyroxine and related compounds in serum and urine by liquid chromatography coupled to triple quadrupole mass spectrometry

Thyroid hormones and their derivatives are structurally related to the non-essential amino acid tyrosine (4-hydroxyphenylalanine). However, there are physicochemical differences that make it difficult to apply an analytical method for their simultaneous detection. This work focused on the optimization of a method using liquid chromatography-electrospray ionization mass spectrometry to measure eight compounds related to levothyroxine (T4). In addition, the influence of the additives to the mobile phase, the solvents for liquid-liquid extraction and the influence of the hydrolysis of the conjugated analytes were studied.Optimization of MRM transitions and collision energy for analytes and capillary voltage, nebulizer gas pressure, nozzle voltage, sheath gas flow, sheath gas temperature, drying gas flow and drying gas temperature for ionization source was done. The recovery of analytes was studied using five solvents and six solvent systems to introduce them into the liquid-liquid extraction and matrix cleanup steps. Different additives to the mobile phase were evaluated as well as the effectiveness of enzymatic and chemical hydrolysis.The best MRM transitions and source parameters were settled in order to generate an optimized instrumental method. The addition of ammonium formate, ammonium fluoride, and acetic acid to the mobile phase showed no improvement in responses compared to classic 0.1% formic acid. The use of tert-butyl methyl ether: isopropanol (75: 25, V: V) showed a suitable recovery of analytes to perform a liquid-liquid extraction, and n-hexane might be an appropriate solvent if a cleanup step is needed. The stability of T3, T4 and thyronine, checked after the hydrolysis with extracts of E. coli and H. pomatia showed good results, contrary to chemical hydrolysis that showed a total degradation of T3 and T4.

CyPA interacts with SERPINH1 to promote extracellular matrix production and inhibit epithelial-mesenchymal transition of trophoblast via enhancing TGF-β/Smad3 pathway in preeclampsia

We previously reported that cyclophilin A (CyPA) production is upregulated in preeclampsia (PE). Moreover, CyPA is known to induce PE-like features in pregnant mice and impair trophoblast invasiveness. In this study, we further illustrated the role of CyPA in PE. RNA-seq analysis, RT-qPCR, immunohistochemical (IHC) staining, and western blotting of mouse placentae revealed that CyPA increased the levels of extracellular matrix (ECM) proteins, such as collagen I and fibronectin, and activated the TGF-β/Smad3 signaling pathway. Additionally, CyPA inhibited the expression of genes involved in epithelial-mesenchymal transition (EMT) (e.g., E-cadherin, N-cadherin, and vimentin) in mouse placentae. We then constructed stable overexpressing and knock-down CyPA cell models (using HTR8/SVneo cells) to clarify the molecular mechanism. We found that CyPA regulated the levels of ECM-related proteins and the EMT process through the TGF-β/Smad3 pathway. We also identified SERPINH1 as a putative CyPA-binding protein, using liquid chromatography-electrospray mass spectrometry (LC-MS)/MS. SERPINH1 was found to be upregulated in the placentae of PE. Silencing SERPINH1 expression reversed the upregulation of ECM proteins and inhibition of the EMT process induced by the overexpression of CyPA. These findings revealed the functions of CyPA in the impaired invasiveness of trophoblasts in PE and indicated that CyPA and SERPINH1 may represent promising targets for the treatment of PE.

Globularia Alypum L. Modulates Inflammatory Markers in the Human Colon and shows a Potential Antioxidant Role in Myeloid Leukemic Cells

Globularia alypum (GA), a plant of the Globulariacea family, has long been used as a traditional cure for inflammatory and metabolic illnesses.In addition to various in vitro model studies, the current work focuses on GA'santioxidant and anti-inflammatory properties in human colon biopsies. Liquid Chromatography-Electrospray Ionization Mass Spectrometry identified the phenol components in GA aqueous extract (GAAE).The antioxidant ability of GAAE was tested in vitro utilizing chemiluminescence and flow cytometry using fluorescent yeastsn conjunction with PLB-985-human myeloid leukemia cells. Experiments on human colon biopsies after a biopsy challenge with Escherichia coli-lipopolysaccharides aimed tosee if GAAE had an anti-inflammatory impact on human colon inflammation.Western blotting was used to assessthe expression of several inflammatory markers. According to the findings, GAAE had a significant influence on hydrogen peroxide and cellular reactive oxygen species.GAAE inhibited the activities of cyclooxygenase 2 and nuclear factor B in inflamed biopsies, indicating anti-inflammatory action.The present study is the first to show that GA has a beneficial effect on human colon inflammation, thanks to its significant antioxidant activity in vitro.According to these preliminary data, GA should be further studies integrative supporting human gastrointestinal inflammatory diseases.

Multi-target bioactivity of summer quinones production in the Persian Gulf burrowing black-type sea urchin

After harvesting the sea urchin gonads for Japanese food “uni” echinoculture systems, the remaining shells and spines are considered waste. However, the material of shells and spines is thought to be rich in natural bioactive molecules. The current study used liquid chromatography–electrospray mass spectrometry to extract summer quinones pigment present in spines and shells of the burrowing sea urchin ‘black’ type Echinometra mathaei from the natural Qeshm Island echinoculture. Then, the biochemical, antioxidant, anti-inflammatory, antidiabetic, antibacterial, and cytotoxic activities of sea urchin quinones pigment were investigated. In terms of bioactivity, both shell and spine pigments demonstrated strong radical scavenging activity (antioxidant). The shell pigment exhibited maximum albumin denaturation inhibition (IC50 = 9.62 μg/ml) (anti-inflammatory), as well as α-amylase inhibition (92.28 percent 4.77) (antidiabetic). Pigments were discovered to have a low antibacterial effect against positive gramme bacteria, as well as low cytotoxic and embryotoxic effects when compared to Artemia salina and zebrafish (Danio rerio). For identification and quantification of pigment extracts, both the photodiode array detector and LC-ESI-MS were used. Spinochrome A, B, and C, as well as echinochrome A, were identified as bioactive quinonoid pigments. This chemical defence is discussed in relation to its algal diet and environmental conditions. In conclusion, the isolated pigments obtained from the shell and spines of E. mathaei sea urchins found to have potent bio-activity and can be used for various biomedical and pharmaceutical applications.

Simultaneous Determination of RDX and HMX in Rat Plasma by LC-MS/MS and its Applications.

Background: 1,3,5-trinitroperhydro-1,3,5-triazine (RDX) and octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX) can cause serious toxicity problems in humans and animals, but direct analyses of RDX and HMX in biological samples are very limited. A rapid and efficient liquid chromatography-electrospray quadrupole linear ion trap mass spectrometry (LC-MS/MS) method suitable for the simultaneous determination of RDX and HMX in rat plasma after intravenous administration of two nitramine compound mixed solutions has been developed. Methods: Plasma samples were pretreated with one-step protein precipitation, the plasma consumption is as low as 100 μl. RDX, HMX, and internal standard mycophenolic acid were eluted for 8.0 min on a reversed-phase C18 analytical column with a water/acetonitrile mixture as the mobile phase. An electrospray ionization (ESI) source was applied and operated in negative ion mode. The optimized mass transition ion pairs (m/z) monitored for RDX, HMX, and internal standard mycophenolic acid were m/z 284.1→61.7, m/z 331.0→108.8, and m/z 319.2→191.1, respectively. Results: The detection ranges of both RDX and HMX in plasma were 5.00–200.00 ng⋅ml−1 with an LOD of 1.00 ng⋅ml−1. The extraction recoveries of RDX and HMX were 60.04 ± 4.18% and 79.57 ± 3.35%, respectively. The precision and accuracy met the requirements, and the method was stable under all tested conditions. Conclusion: The present method is miniaturized, effective, portable, rapid and can be easily used for simultaneous quantification of RDX and HMX in rat plasma.

Anti-inflammatory and anti-aggregating effects of rangpur in the first trimester of growth: ultra-performance liquid chromatography-electrospray mass spectrometry profile and quantification of hesperidin.

Citrus fruits are a rich source of valuable molecules, and their industrial processing produces bagasses, little explored to generate important by-products. These Citrus residues, including seeds and peels, also contain numerous pharmacologically important substances. To reduce the impact of these Citrus by-products, young, harvested fruits could be used as a functional supplemental food while another part is grown until maturity for industrial production. This study therefore aims to valorize rangpur (Citrus limonia) in the first 3 months of its growth by investigating and comparing its monthly chemical profiles using ultra-performance liquid chromatography-electrospray mass spectrometry (UPLC-ESI-MS) and its anti-inflammatory and antiplatelet activity. Extracts obtained from the fruits harvested in November, December, and January, 2017 and 2018 (L221117, L161217, and L160118) showed different UPLC-ESI-MS profiles. Twenty-five of the 26 detected metabolites were identified as cyclitol, pyrrolidine betaine, aryl propanoyl esters, chlorogenic acids, flavonoids, coumarins, and limonoids. Quantification studies indicated an increased concentration of hesperidin from the younger fruits to the older fruits of the series. L160118 reduced nitrogen oxide (NOx), tumor necrosis factor alpha (TNF-α), and interleukin 6 (IL-6) levels more than other extracts. Their activity followed the same trends as the hesperidin concentration in each fruit. In contrast, the most promising antiplatelet activity was observed with the extracts from the two youngest fruits. This suggests combined effects of the chemical components found in these fruits' extracts. The extracts obtained from these young fruits showed considerable anti-inflammatory and antiplatelet activity. Overall, young rangpur could be used as raw material to produce functional foods without producing any waste. © 2022 Society of Chemical Industry.

Phyto-Beneficial Traits of Rhizosphere Bacteria: In Vitro Exploration of Plant Growth Promoting and Phytopathogen Biocontrol Ability of Selected Strains Isolated from Harsh Environments

Beneficial interactions between plants and some bacterial species have been long recognized, as they proved to exert various growth-promoting and health-protective activities on economically relevant crops. In this study, the growth promoting and antifungal activity of six bacterial strains, Paenarthrobacter ureafaciens, Beijerinckia fluminensis, Pseudomonas protegens, Arthrobacter sp., Arthrobacter defluii, and Arthrobacter nicotinovorans, were investigated. The tested strains resulted positive for some plant growth promoting (PGP) traits, such as indole-3-acetic acid (IAA), 1-aminocyclopropane-1-carboxylate-deaminase (ACC-deaminase), siderophore production, and solubilization of phosphates. The effect of the selected bacteria on Arabidopsis thaliana seedlings growth was assessed using different morphological parameters. Bacterial activity against the phytopathogenic fungal species Aspergillus flavus, Fusarium proliferatum, and Fusarium verticillioides was also assessed, since these cause major yield losses in cereal crops and are well-known mycotoxin producers. Strains Pvr_9 (B. fluminensis) and PHA_1 (P. protegens) showed an important growth-promoting effect on A. thaliana coupled with a high antifungal activity on all the three fungal species. The analysis of bacterial broths through ultra performance liquid chromatography–mass spectrometry (UPLC–MS) and liquid chromatography–electrospray ionization–mass spectrometry (LC–ESI–MS/MS) confirmed the presence of potential PGP-compounds, among these are desferrioxamine B, aminochelin, asperchrome B, quinolobactin siderophores, and salicylic acid.

Phosphorylation and glycosylation isoforms of bovine κ-casein variant E in homozygous Swedish Red cow milk detected by liquid chromatography-electrospray ionization mass spectrometry

Variations in the phosphorylation and glycosylation patterns of the common κ-casein (CN) variants A and B have been explored, whereas studies on variant E heterogeneity are scarce. This study reports for the first time the detailed phosphorylation and glycosylation pattern of the κ-CN variant E in comparison with variants A and B. Individual cow milk samples representing κ-CN genotype EE (n = 12) were obtained from Swedish Red cows, and the natural posttranslational modifications of its κ-CN were identified and quantified by liquid chromatography-electrospray mass spectrometry. In total, 12 unique isoform masses of κ-CN variant E were identified. In comparison, AA and BB milk consisted of 14 and 17 unique isoform masses, respectively. The most abundant κ-CN E isoform detected in the EE milk was the monophosphorylated, unglycosylated [1P 0G, ∼70%; where P indicates phosphorylation from single to triple phosphorylation (1-3P), and G indicates glycosylation from single to triple glycosylation (1-3G)] form, followed by diphosphorylated, unglycosylated (2P 0G, ∼12%) form, resembling known patterns from variants A and B. However, a clear distinction was the presence of the rare triphosphorylated, nonglycosylated (3P 0G, ∼0.05%) κ-CN isoform in the EE milk. All isoforms detected in variant E were phosphorylated, giving a phosphorylation degree of 100%. This is comparable with the phosphorylation degree of variants A and B, being also almost 100%, though with very small amounts of nonphosphorylated, glycosylated isoforms detected. The glycosylation degree of variant E was found to be around 17%, a bit higher than observed for variant B (around 14%), and higher than variant A (around 7%). Among glycosylation, the glycan e was the most common type identified for all 3 variants, followed by c/d (straight and branched chain trisaccharides, respectively), and b. In contrast to κ-CN variants A and B, no glycan of type a was found in variant E. Taken together, this study shows that the posttranslational modification pattern of variant E resembles that of known variants to a large extent, but with subtle differences.

Investigation by top‐down high‐performance liquid chromatography–mass spectrometry of glutathionylation and cysteinylation of salivary S100A9 and cystatin B in preterm newborns

AbstractGlutathionylation and cysteinylation can be involved in the protection of critical cysteines from irreversible oxidative damages. S100A9 long and cystatin B, proteins highly represented in the saliva of preterm and at‐term newborns, can undergo these modifications. Levels of S100A9 long and cystatin B and their glutathionylated and cysteinylated derivatives have been determined by a top‐down platform based on high‐performance liquid chromatography–electrospray ionization–mass spectrometry in 100 salivary samples serially collected from 17 preterm newborns with different postconceptional age at birth (178‐226 days) and in 90 salivary samples collected from at‐term newborns and babies. Results showed that: (1) S100A9 long and cystatin B were mainly present as unmodified forms in extremely preterm newborn; (2) the percentage of the S‐thiolated derivatives of both proteins increased with increasing the postconceptional age; (3) the greatest variation occurred up to about 280 days of postconceptional age. Interestingly, differences in the levels of the S‐thiolated derivatives only depended on the postconceptional age and not on whether the infant was born preterm or at‐term. Inadequate levels of cysteine and glutathione might be responsible for the low level of S‐thiolated derivatives measured in preterm newborns. Data are available via ProteomeXchange with identifier PXD025517.

Bioactive compounds of parsley (Petroselinum crispum), chives (Allium schoenoprasum L) and their mixture (Brazilian cheiro-verde) as promising antioxidant and anti-cholesterol oxidation agents in a food system

This study determined the bioactive composition and antioxidant potential of parsley, chives and their mixture (Brazilian cheiro-verde). Additionally, the effect of these herbs against cholesterol oxidation in grilled sardines (Sardinella brasiliensis) was also investigated. Ultra-high Performance Liquid Chromatography-Electrospray Ionization-Mass Spectrometry (UHPLC-ESI-MS) analyses revealed the presence of phenolic acids (caffeic, chlorogenic, and ferulic acids) and flavonoids (apigenin, kaempferol, catechin) in the herbs. Higher levels of phenolics (2.10 ± 0.02 mg GAE/g) and carotenoids (205.95 ± 0.17 µg/g) were determined in parsley extracts. Moreover, parsley also presented higher antioxidant capacity by DPPH (59.21 ± 0.07 %) and ORAC (109.94 ± 18.7 µM TE/g) than the other herbs. In vivo analyses demonstrated that the herbs’ extracts decreased the damage on Saccharomyces cerevisiae cells exposed to H2O2, except the chives extract at 10 μg/mL. Higher levels of cholesterol oxidation products (COPs) were determined after grilling. The total COPs increased from 61.8 ± 0.7 (raw fish) to 139.7 ± 10.1 µg/g (control). However, the addition of herbs effectively reduced cholesterol oxides formation, this effect was more pronounced in fish containing 4% parsley and 4% cheiro-verde. Promising results were found for cheiro-verde; however, it did not present synergic antioxidant effects.

Effects of Fengycins and Iturins on Fusarium oxysporum f. sp. physali and Root Colonization by Bacillus velezensis Bs006 Protect Golden Berry Against Vascular Wilt.

Bacillus velezensis Bs006 has shown antagonistic activity on Fusarium oxysporum f. sp. physali and biocontrol activity against Fusarium wilt (FW) in golden berry (Physalis peruviana). We hypothesized that strain Bs006 has the ability to synthesize antimicrobial cyclic lipopeptides (CLPs) like other members of the same species. However, if so, the real effects of CLPs on F. oxysporum f. sp. physali and their potential as a biocontrol tool against Physalis-FW have not been elucidated. In this study the CLPs profile of Bs006 in liquid culture and antagonist-plant-pathogen interactions were characterized. Also, the potential effects of supernatant free of bacteria against F. oxysporum f. sp. physali and FW were explored and compared with the effects of pure CLPs. Ultraperformance liquid chromatography-electrospray ionization-mass spectrometry analysis revealed the capacity of Bs006 to synthesize homologous compounds of iturins, surfactins, and fengycins in liquid culture and on the inhibition zone against F. oxysporum f. sp. physali in dual confrontation tests. Bs006 supernatant reduced the germination and growth of F. oxysporum f. sp. physali and caused vacuolization, swelling, and lysis of F. oxysporum f. sp. physali cells in a concentration-dependent manner. Pure fengycins affected the development of F. oxysporum f. sp. physali from 11 mg/liter and iturins from 21 mg/liter. In a gnotobiotic system, Bs006 colonized the root surface of golden berry, inhibited the growth of F. oxysporum f. sp. physali, and produced CLPs. Individual application of Bs006 and supernatant protected the plants from F. oxysporum f. sp. physali infections by 37 to 53%, respectively. Meanwhile, fengycins reduced the disease progress by 39%. These results suggest further studies to select an optimum combination of Bs006 and supernatant or CLPs, which might be a good option as biofungicide against F. oxysporum f. sp. physali.

Abstract 12998: S-Cysteinylation of Plasma and Serum Albumin is Associated With Risk Factors of Atherosclerotic Cardiovascular Disease

Introduction: Albumin oxidation or S-cysteinylation (S-Cys) of its free cysteine residue has been shown to be a measure of oxidative stress. While it is subject to artificial ex-vivo elevations, freshly processed samples could serve as a theoretical biomarker of any number of inflammatory states. Hypothesis: S-Cys Albumin is associated with atherosclerotic cardiovascular disease (ASCVD) and its risk factors that increase oxidative stress. Methods: We enrolled 137 patients evaluated for chest pain in a single-center cross-sectional study from May 2016 through March 2020. Plasma and serum samples were gathered and immediately processed for analysis of intact albumin by high pressure liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS). Statistical analysis included a spearman’s coefficient, and two-tailed linear regression of the relative fractional abundance (RFA) of S-Cys albumin and clinical variables. Results: S-Cys-albumin levels were significantly decreased in those with a previous history of coronary stenting (Plasma β=-0.032, p=0.005; Serum β=-0.032, p=0.012; n=24) and MDD (Plasma β=-0.016, p=0.132; Serum β=-0.026, p=0.021, n=34). S-Cys-albumin levels were significantly elevated in those with a previous history of peripheral artery disease (Plasma β=0.051, p=0.015; Serum β=0.055, p= 0.016; n=6) and chronic infections (Plasma β=0.023, p=0.033; Serum β=0.021, p =0.069; n=30). CAD by history was not significantly different (Plasma β=0.012, p=0.19; Serum β=0.01, p =0.3; n=57). Spearman’s coefficient demonstrated a positive correlation between age (plasma rho = 0.1877, p = 0.0576; serum rho 0.2641, p = 0.007; n=103), BMI (plasma rho = 0.2089, p = 0.0342; serum rho = 0.1287, p = 0.195; n=103), Framingham risk score (plasma rho = 0.2607, p = 0.0523; serum rho = 0.3595, p = 0.0065; n=56), ASCVD risk score (plasma rho = 0.3526, p = 0.0083; serum rho = 0.472, p = 0.0003; n=55), heart rate (plasma rho = 0.2225, p = 0.0239, serum rho = 0.1678, p = 0.0902; n=103) and NT-pro-BNP (plasma rho = 0.3398, p = 0.0319; serum rho 0.3937, p = 0.012; n=40). Conclusion: S-Cysteinylated Albumin is associated with several measure of ASCVD and its risk factors. It may be a marker of the underlying oxidative stress.

Comprehensive analysis of natural polysaccharides from TCMs: a generic approach based on UPLC-MS/MS

Here, we report a new application using liquid chromatography-electrospray mass spectrometry (UHPLC-ESI-MS) using aldononitrile acetate derivatives for simultaneous baseline separation and detection of eight neutral saccharides, two uronic acids, one ketose, and eight alditols within 14 min. The separation was performed on a Cortecs C₁₈ column using acetonitrile (A) and water (B) as the mobile phase with gradient elution. The target components were detected in selected ion monitoring (SIM) mode by mass spectrometry with an electrospray ionization (ESI) source operating in positive ionization mode. A comparison with traditional methods was used to determine the validity of the results. The UHPLC-ESI-MS method was used for quantitative analysis of free carbohydrates in water extracts of Crataegus pinnatifida as well as determination of Polygonatum cyrtonema and Glossy ganoderma monosaccharides in polysaccharides. The results demonstrate that this protocol is a comprehensive and effective technique for qualitative and quantitative analysis of plant polysaccharides from TCMs.

Predicting compound amenability with liquid chromatography-mass spectrometry to improve non-targeted analysis.

With the increasing availability of high-resolution mass spectrometers, suspect screening and non-targeted analysis are becoming popular compound identification tools for environmental researchers. Samples of interest often contain a large (unknown) number of chemicals spanning the detectable mass range of the instrument. In an effort to separate these chemicals prior to injection into the mass spectrometer, a chromatography method is often utilized. There are numerous types of gas and liquid chromatographs that can be coupled to commercially available mass spectrometers. Depending on the type of instrument used for analysis, the researcher is likely to observe a different subset of compounds based on the amenability of those chemicals to the selected experimental techniques and equipment. It would be advantageous if this subset of chemicals could be predicted prior to conducting the experiment, in order to minimize potential false-positive and false-negative identifications. In this work, we utilize experimental datasets to predict the amenability of chemical compounds to detection with liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS). The assembled dataset totals 5517 unique chemicals either explicitly detected or not detected with LC-ESI-MS. The resulting detected/not-detected matrix has been modeled using specific molecular descriptors to predict which chemicals are amenable to LC-ESI-MS, and to which form(s) of ionization. Random forest models, including a measure of the applicability domain of the model for both positive and negative modes of the electrospray ionization source, were successfully developed. The outcome of this work will help to inform future suspect screening and non-targeted analyses of chemicals by better defining the potential LC-ESI-MS detectable chemical landscape of interest.

Comprehensive structural characterisation of the newly emerged synthetic cannabimimetics Cumyl-BC[2.2.1]HpMeGaClone, Cumyl-BC[2.2.1]HpMINACA, and Cumyl-BC[2.2.1]HpMICA featuring a norbornyl methyl side chain

Synthetic cannabimimetics (SCs) are a broad group of new psychoactive substances (NPS) that make up approximately a quarter of all NPS on the European drug market, and new variants continue to emerge. As legislation in many countries evolves, the variety of SCs correspondingly grows, making SCs a persistent phenomenon that continues to be relevant. Here, we describe the first identification and comprehensive analytical characterisation of three SCs from seized case samples and test purchases that newly emerged in 2020/2021, and we discuss analytical differences crucial for identification. Structure elucidation and analytical characterisation were performed within the EU-project ADEBAR11Ausbau analytischer Datenbanken, Erhebung und bundesweite Bereitstellung von analytischen Daten, pharmakologischen Daten, Verfahrensvorschriften und Referenzmaterialien für neu auf dem Drogenmarkt auftretende Stoffe (engl. expansion of analytical databases through collection and nationwide publication of analytical and pharmacological data and the distribution of standard operating procedures as well as reference materials for substances newly appearing on the drug market).plus using gas chromatography-mass spectrometry (GC–MS), gas chromatography-solid state infrared spectroscopy (GC-sIR), as well as solid and neat IR spectroscopy, Raman spectroscopy, liquid chromatography-electrospray ionisation-mass spectrometry (LC-ESI-MS), and high resolution (HR)-LC-ESI-MS, and nuclear magnetic resonance (NMR) spectroscopy. The raw analytical data have immediately been published in printed format and electronic formats by the authors on the European Database on New Drugs (EDND), to all German Police and Customs laboratories, and on the internationally available internet platform NPS datahub (www.nps-datahub.com). Endo- and exo-isomers can be differentiated by IR and NMR and were separated using a standard GC temperature program. Quantum-chemistry-based calculations of the NMR spectra were executed to distinguish between the endo- and exo-isomers of the SCs in mixtures. After identification of these new compounds, the EU-project ADEBAR plus initialised the update of the group definitions in the German New Psychoactive Substance Act (NpSG) to include the norbornyl methyl (NBM) side chain.

An analysis of urine and serum amino acids in critically ill patients upon admission by means of targeted LC\u2013MS/MS: a preliminary study

Sepsis, defined as a dysregulated host response to infection, causes the interruption of homeostasis resulting in metabolic changes. An examination of patient metabolites, such as amino acids, during the early stage of sepsis may facilitate diagnosing and assessing the severity of the sepsis. The aim of this study was to compare patterns of urine and serum amino acids relative to sepsis, septic shock and survival. Urine and serum samples were obtained from healthy volunteers (n = 15) once or patients (n = 15) within 24 h of a diagnosis of sepsis or septic shock. Concentrations of 25 amino acids were measured in urine and serum samples with liquid chromatography-electrospray mass spectrometry. On admission in the whole cohort, AAA, ABA, mHis, APA, Gly-Pro and tPro concentrations were significantly lower in the serum than in the urine and Arg, Gly, His, hPro, Leu, Ile, Lys, Orn, Phe, Sarc, Thr, Tyr, Asn and Gln were significantly higher in the serum than in the urine. The urine Gly-Pro concentration was significantly higher in septic shock than in sepsis. The serum Cit concentration was significantly lower in septic shock than in sepsis. The urine ABA, mHis and Gly-Pro, and serum Arg, hPro and Orn concentrations were over two-fold higher in the septic group compared to the control group. Urine and serum amino acids measured in septic patients on admission to the ICU may shed light on a patient’s metabolic condition during sepsis or septic shock.

Is selenium beneficial or detrimental to earthworm? Growth and metabolism responses of Eisenia Fetida to Na2SeO3 exposure

Se unevenly distributed in soils due to variations of geology and anthropogenic input, which results in different effects on earthworms. The effects of Se were characterized by analyzing the growth and metabolism responses of earthworms after exposure to three different concentrations of Na2SeO3. The results showed that except the possible growth promotion at 5 mg/kg, low and middle-level exposure to Na2SeO3 (0.3–10 mg/kg) did not significantly affect the growth of earthworms. While a significant inhibition effect on growth was observed in the high-level exposure group (30–70 mg/kg). There was an inflection point for Se performing promotion to inhibition effects on earthworm growth. To investigate the metabolic response of earthworms, a novel HPLC-ESI-MS (High Performance Liquid Chromatography-Electrospray Ionization-Mass Spectrometry) method was used to determine sensitive biomarkers. Selenium exposure significantly altered the metabolism of seven essential amino acids, namely tyrosine, leucine, phenylalanine, valine, alanine, glycine, and lysine, and two selenoamino acids, namely selenomethionine and methylselenocysteine. The overall metabolism level of earthworms was not affected at low exposure concentrations, but was affected at medium and high exposure concentrations. The metabolic pathways that integrated the selenocompound metabolism and the tricarboxylic acid cycle from the perspective of energy supply and demand were affected by Na2SeO3 exposure. The derived reactive oxygen species at high exposure concentrations were probably the reason for the growth inhibition effect of Se on earthworms. This study provides biochemical insights into the effects of Na2SeO3 on earthworms and suggests that an Se concentration of about 2.3 mg/kg is appropriate for soil organism health.

Cytotoxic Properties, Anthocyanin and Furanocoumarin Content of Red-Pigments Obtained from Callistemon citrinus (Curtis) Skeels Flowers

Background and objectives: There is growing interest in introducing safe and bioactive natural red pigments to the pharmaceutical and cosmetic industries. This study was designed to determine the phytochemical content and potential cytotoxicity of red pigment from Callistemon citrinus (Curtis) Skeels (syn. Melaleuca citrina (Curtis) Dum.Cours.) flowers. Methods: The flowers’ anthocyanin rich pigment was extracted with ethanol (70%, v/v) containing 0.5% formic acid. This extract was fractionated by a three-step process through a Sep-Pak C18 cartridge with water, ethyl acetate, and methanol, respectively. The anthocyanin and coumarin content of the red pigment was identified based on a reverse phase high performance liquid chromatography-electrospray ionization mass spectrometry (HPLC/ESI-MS/MS). The MTT assay was used to assess the cytotoxicity of red pigment evaluated on normal human foreskin fibroblasts as well as two malignant cell lines: human breast cancer (MCF-7) and human fibrosarcoma cells (HT1080) at 24, 48 and 72 hours. Results: The anthocyanin compounds in the red pigment fraction were cyanidin (1), cyanidin 3,5-O-diglucoside (2), cyanidin 3-O-glucoside (3), and cyanidin 3-O-glucoside-8-ethyl-catechin (4). Flowers’ pigment also contained two furanocoumarins, including 8-(but-2-en-2-yl)-8,9-dihydro-2H-furo[2,3-h] chromen-2-one (or 15-methyl angenomallin, (5)) and 9-methyl-7H-furo[3,2-g] chromen-7-one (or 8-methylpsoralen, (6)). According to the MTT assays, the highest cytotoxic effect was observed on human foreskin fibroblasts with an IC50 values of <12.5 (µg/ mL, 24 h) and 85.2 (µg/ mL, 48 h). Conclusion: It might be assumed that application of the red pigment of C. citrinus in topical formulations and cosmetics should be done with caution due to the observed cytotoxicity on dermal fibroblasts.

Mitochondrial Disruption by Amyloid Beta 42 Identified by Proteomics and Pathway Mapping.

Alzheimer’s disease (AD) is marked by chronic neurodegeneration associated with the occurrence of plaques containing amyloid β (Aβ) proteins in various parts of the human brain. An increase in several Aβ fragments is well documented in patients with AD and anti-amyloid targeting is an emerging area of therapy. Soluble Aβ can bind to various cell surface and intracellular molecules with the pathogenic Aβ42 fragment leading to neurotoxicity. Here we examined the effect of Aβ42 on network adaptations in the proteome of nerve growth factor (NGF) differentiated PC12 cells using liquid-chromatography electrospray ionization mass spectrometry (LC-ESI MS/MS) proteomics. Whole-cell peptide mass fingerprinting was coupled to bioinformatic gene set enrichment analysis (GSEA) in order to identify differentially represented proteins and related gene ontology (GO) pathways within Aβ42 treated cells. Our results underscore a role for Aβ42 in disrupting proteome responses for signaling, bioenergetics, and morphology in mitochondria. These findings highlight the specific components of the mitochondrial response during Aβ42 neurotoxicity and suggest several new biomarkers for detection and surveillance of amyloid disease.