Quadrupole Ion Research Articles

In vitro assessment of the bioavailability of bioactive non-extractable polyphenols obtained by pressurized liquid extraction combined with enzymatic-assisted extraction from sweet cherry (Prunus avium L.) pomace

In vitro digestion and absorption simulation processes of non-extractable polyphenols (NEPs) obtained by pressurized liquid extraction combined with enzymatic-assisted extraction with Promod enzyme (PLE-EAE) from the residue of conventional extraction of sweet cherry pomace were studied. In general, total phenolic and proanthocyanidin contents decreased in each phase of the digestion. However, the antioxidant capacity increased when the digestion process progressed. In addition, the highest total phenolic and proanthocyanidin contents and antioxidant capacity were obtained in the absorbed fraction. NEPs from PLE-EAE extract, digestive fractions, absorbed and unabsorbed fractions were analyzed by ultra-high-performance liquid chromatography coupled to electrospray ionization quadrupole Exactive-Orbitrap mass spectrometry (UHPLC-ESI-Q-Orbitrap-MS). Fifteen NEPs were identified in the intestinal fraction and five in the absorbed fraction after the digestion process. Results obtained in this study define for the first time the bioavailability of antioxidant NEPs obtained from sweet cherry pomace.

Identification of cadmium containing metabolites in HepG2 cells after treatment with cadmium-selenium quantum dots

The transformation of quantum dots (QDs) by organisms has attracted broad attention but remains unclear. Understanding of the metabolites helps to reveal the transformation pathway of QDs. Cd containing-metallothionein (MT) are the main species formed by Cd released from CdSe QDs in HepG2 cells, while speciation analysis of Cd containing MTs remains a challenge because MTs has several subisoforms and can bind with several metals. Herein, we built a hyphenated platform for speciation analysis of QDs in HepG2 cells after treatment with CdSe/ZnS QDs. The Cd-containing MTs were separated in reversed phase high performance liquid chromatography (RP-HPLC) and subsequently online detected by inductively coupled plasma mass spectrometry (ICP-MS) and electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-Q-TOF-MS) parallelly. Four groups of Cd-containing metabolites were found by detecting Cd in ICP-MS. Their structures were identified in ESI-Q-TOF-MS and further confirmed with standards of four subisoforms of MT, including N-terminal acetylation MT2a, N-terminal acetylation MT1e, N-terminal acetylation MT1g and MT1m. Each group of them contains various stoichiometry of Cd/Zn. The metabolites of QDs remain same while the concentrations of each metabolite and its stoichiometry of Cd/Zn vary for different incubation concentration/time. This work provides a new parallel hyphenation technique of HPLC-ICP-MS/ESI-MS with high separation resolution and powerful detection ability, and the obtained results provide detailed metabolism information of QDs in HepG2 cells after treatment of CdSe/ZnS QDs, contributing to deep exploration of the functional mechanisms of QDs in organisms.

Navigating through the Lipid Metabolism Maze: Diagnosis and Prognosis Metabolites of Hepatocellular Carcinoma versus Compensated Cirrhosis.

(1) Background: The pursuit of finding biomarkers for the diagnosis and prognosis of hepatocellular carcinoma (HCC) has never been so paramount in the days of personalized medicine. The main objective of our study is to identify new biomarkers for diagnosing HCC, and to identify which patients are at risk of developing tumor recurrence, decompensation, or even possesses the risk of cancer-related death. (2) Methods: We have conducted an untargeted metabolomics study from the serum of 69 European patients—32 compensated cirrhotic patients without HCC (controls), and 37 cirrhotic patients with HCC with compensated underlying liver disease (cases), that underwent curative treatment (surgery or ablation), performing ultra-high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry (UHPLC-QTOF- (ESI+)-MS) with an emphasis on lipid metabolites. (3) Results: 1,25-dihydroxy cholesterol (m/z = 419.281), myristyl palmitate (m/z = 453.165), 25-hydroxy vitamin D2 (m/z = 413.265), 12-ketodeoxycholic acid (m/z = 391.283), lysoPC (21:4) (m/z = 558.291), and lysoPE (22:2) (m/z = 534.286) represent notable biomarkers that differentiate compensated cirrhosis from early HCC, and ceramide species are depleted in the serum of HCC patients. Regarding prognosis, no metabolite identified in our study could determine tumor relapse. To distinguish between the HCC patients that survived curative treatment and those at risk that developed tumor burden, we have identified two notable phosphocholines (PC (30:2); PC (30:1)) with AUROCs of 0.820 and 0.807, respectively, that seem to increase when patients are at risk. In a univariate analysis, arachidonic acid was the only metabolite to predict decompensation (OR = 0.1, 95% CI: 0–0.16, p < 0.005), while in the multivariate analysis, dismally, no variable was associated with decompensation. Furthermore, in the multivariate analysis, we have found out for the first time that the increased expression of 1,25-dihydroxy cholesterol, myristyl palmitate, 12-keto deoxycholic acid, lysoPC (21:4), and lysoPE (22:2) are independent markers of survival. (4) Conclusions: Our study reveals that lipids play a crucial role in discriminating compensated cirrhosis and early hepatocellular carcinoma, and might represent markers of survival and prognosis in personalized and minimally invasive medicine.

Comprehensive characterization and identification of chemical constituents in Yangwei decoction using ultra high performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry.

Yangwei decoction, a classical traditional Chinese medicine prescription, has been widely used to treat exogenous cold and internal injury with damp stagnation for many centuries. However, its systematic chemical profiling remains ambiguous, which has hampered the interpretation of pharmacology and the mechanism of its formula. In the present study, ultra-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry method was successfully established for the first time to separate and identify the complicated components of Yangwei decoction. The accurate mass data of the protonated molecules, deprotonated molecules, and fragment ions were detected in positive and negative ion modes. A total of 226 compounds in Yangwei decoction were tentatively identified and unambiguously characterized by comparing their retention times and mass spectrometry data with those of reference standards and literature, including 24 lignans, 18 alkaloids, 9 phenylpropanoid glycosides, 76 flavonoids, 59 triterpenoids, 17 organic acids, 7 gingerols, 8 lactones, and 8 other compounds. The present study provides a novel method of constituents characterization for well-known Chinese medicine prescriptions. The study aims to lay a robust foundation for future research, providing the holistic quality control and pharmacology of Yangwei decoction.

UHPLC-ESI-QTOF-MS2 analysis of Acacia pennata extract and its effects on glycemic indices, lipid profile, pancreatic and hepatorenal alterations in nicotinamide/streptozotocin-induced diabetic rats.

Diabetes mellitus (DM) is a chronic disorder associated with severe metabolic derangement and comorbidities. The constant increase in the global population of diabetic patients coupled with some prevailing side effects associated with synthetic antidiabetic drugs has necessitated the urgent need for the search for alternative antidiabetic regimens. This study investigated the antidiabetic, antioxidant, and pancreatic protective effects of the Acacia pennata extract (APE) against nicotinamide/streptozotocin induced DM in rats. The antidiabetic activity of APE was evaluated and investigated at doses of 100 and 400 mg/kg body weight, while metformin (150 mg/kg bw) was used as a standard drug. APE markedly decreased blood glucose level, homeostatic model assessment for insulin resistance, serum total cholesterol, triglycerides, low‐density lipoprotein, blood urea nitrogen, creatinine, alanine transaminase, aspartate transaminase, and alanine phosphatase levels. Additionally, treatment with APE increased the body weight, serum insulin concentration, and high‐density lipoprotein. Moreover, activities of pancreatic superoxide dismutase, catalase, and glutathione peroxidase were increased, while the altered pancreatic architecture in the histopathological examination was notably restored in the treated rats. Ultra‐high performance liquid chromatography combined with electrospray ionization quadrupole time‐of‐flight mass spectrometry (UHPLC‐ESI‐QTOF‐MS) analysis of APE showcases the prevailing presence of polyphenolic compounds. Conclusively, this study showed the beneficial effects of the Acacia pennata in controlling metabolic derangement, pancreatic and hepatorenal dysfunction in diabetic rats.

Analytical Strategies for LC-MS-Based Untargeted and Targeted Metabolomics Approaches Reveal the Entomological Origins of Honey.

A comprehensive liquid chromatography-mass spectrometry (LC-MS)-based metabolomics approach was developed to discriminate honey harvested from Apis mellifera ligustica Spinola (A. mellifera) and Apis cerana cerana Fabricius (A. cerana). Based on an untargeted strategy, ultrahigh-performance liquid chromatography electrospray ionization quadrupole orbitrap high-resolution mass spectrometry (UPLC Q-Orbitrap) was combined with chemometrics techniques to screen and identify tentative markers from A. mellifera and A. cerana honey. In targeted metabolomics analysis, a sensitive method of solid-phase extraction followed by ultrahigh-performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry (UPLC-MS/MS) was established for quantifying three markers, and the results showed that 3-amino-2-naphthoic acid and methyl indole-3-acetate could be considered markers of A. cerana honey, as they were present in higher amounts in A. cerana honey than in A. mellifera honey, whereas kynurenic acid was determined to be a marker of A. mellifera honey. This work highlights critical information for the authentication of A. cerana and A. mellifera honey.

Integration of metabolomics and transcriptomics data to further characterize Gliricidia sepium (Jacq.) Kunth under high salinity stress

Soil salinity is one abiotic stress that threatens agriculture in more than 100 countries. Gliricidia [Gliricidia sepium (Jacq.) Kunth] is a multipurpose tree known for its ability to adapt to a wide range of soils; however, its tolerance limits and responses to salt stress are not yet well understood. In this study, after characterizing the morphophysiological responses of young gliricidia plants to salinity stress, leaf metabolic and transcription profiles were generated and submitted to single and integrated analyses. RNA from leaf samples were subjected to RNA sequencing using an Illumina HiSeq platform and the paired-end strategy. Polar and lipidic fractions from leaf samples were extracted and analyzed on an ultra-high-performance liquid chromatography (UHPLC) coupled with electrospray ionization quadrupole time-of-flight high-resolution mass spectrometry (MS) system. Acquired data were analyzed using the OmicsBox, XCMS Online, MetaboAnalyst, and Omics Fusion platforms. The substrate salinization protocol used allowed the identification of two distinct responses to salt stress: tolerance and adaptation. Single analysis on transcriptome and metabolome data sets led to a group of 5,672 transcripts and 107 metabolites differentially expressed in gliricidia leaves under salt stress. The phenylpropanoid biosynthesis was the most affected pathway, with 15 metabolites and three genes differentially expressed. Results showed that the differentially expressed metabolites and genes from this pathway affect mainly short-term salt stress (STS). The single analysis of the transcriptome identified 12 genes coding for proteins that might play a role in gliricidia response at both STS and long-term salt stress (LTS). Further studies are needed to reveal the mechanisms behind the adaptation response.

Investigation of the Variability of Alkaloids in Buxus sempervirens L. Using Multivariate Data Analysis of LC/MS Profiles.

Buxus sempervirens L. is a common ornamental plant in southern and central Europe, and has been used ethopharmacologically against a wide variety of diseases due to it containing nor-triterpene alkaloids of the nor-cycloartane type. Recently, we demonstrated the interesting antiprotozoal potential of some of these compounds. To characterize the temporal variability in the alkaloid profile of two different varieties and their leaves and twigs, 30 different extracts of B. sempervirens were evaluated by Ultra High Performance Liquid Chromatography/positive Mode-Electrospray Ionization Quadrupole Time-of-Flight-Tandem Mass Spectrometry (UHPLC/+ESI-QqTOF-MS/MS). The analytical profiles were thoroughly investigated by various methods of multivariate data analysis (MVDA). A principal component analysis (PCA) model elucidates the seasonal variation in the phytochemical composition of B. sempervirens var. arborescens and suffruticosa along with differences between the varieties. Analysis of a volcano plot illustrated the differences between the two organs, the leaf and twig. Eighteen compounds were highlighted by the models as constituents of the plant characteristic for a season, variety or organ. These compounds were dereplicated based on their chromatographic and +ESI-QqTOF-MS and –MS/MS data. In addition, mass spectral fragmentation pathways for already known alkaloids as well as new natural products could be postulated for the first time. In conclusion, the MVDA models give detailed information on the temporal variability in the alkaloid profile of two different varieties and their organs (leaf vs. twig) of B. sempervirens. Thus, the results of this study allow, e.g., the identification of characteristic compounds for the different varieties, plant organs, seasons, and the optimal harvesting time for the isolation of particular Buxus-alkaloids of interest for subsequent studies.

Synthesis and analytical differentiation of a novel synthetic cathinone 1-(2,3-dihydro-1H-inden-5-yl)-2-(pyrrolidin-1-yl)butan-1-one (5-PPDI) and its regioisomers

The differentiation of a controlled drug from uncontrolled novel compounds related to regioisomers is a significant issue in forensics. The compound 1-(2,3-dihydro-1H-inden-5-yl)-2-(pyrrolidin-1-yl)butan-1-one (5-PPDI) is a novel synthetic cathinone first identified in 2015 as a new psychoactive substance. 5-PPDI is the indanyl analog of α-PBP, a typical synthetic cathinone, as well as the aliphatic ring analog of MDPBP, a synthetic cathinone with a heterocyclic ring. 5-PPDI can have three possible regioisomers, 1-(2,3-dihydro-1H-inden-1-yl)-2-(pyrrolidin-1-yl)butan-1-one (1-PPDI), 1-(2,3-dihydro-1H-inden-2-yl)-2-(pyrrolidin-1-yl)butan-1-one (2-PPDI), and 1-(2,3-dihydro-1H-inden-4-yl)-2-(pyrrolidin-1-yl)butan-1-one (4-PPDI), differing by the substituted position of the alkyl side-chain. In this study, 5-PPDI and its three regioisomers were synthesized from commercially available precursor compounds and analyzed by gas chromatography/electron ionization–quadrupole mass spectrometry (GC/EI–Q-MS) and by liquid chromatography/electrospray ionization–linear ion trap mass spectrometry (LC/ESI–LIT–MS). The four PPDI isomers could be differentiated by GC/EI–Q-MS with three types of separation columns and by 2nd generation product ion spectra (MS3) analysis using ESI–LIT–MS.

Assessment of the Phytochemical and Nutrimental Composition of Dark Chia Seed (Salvia hispánica L.).

Chia seeds are rich sources of different macro and micronutrients associated with health benefits; thus, they may be considered as a functional food. However, the composition depends on the variety, origin, climate and soil. Here, we show a comprehensive characterization of extractable and non-extractable phenolic compounds of dark chia seed Salvia hispanica L. using high-performance liquid chromatography–electrospray ionization–quadrupole time-of-flight (HPLC-ESI-QTOF) and discuss potential health benefits associated with the presence of a number of nutritional and bioactive compounds. We report that dark chia from Jalisco is a high-fiber food, containing omega-3 polyunsaturated fatty acids, essential amino acids (phenylalanine and tryptophan), and nucleosides (adenosine, guanidine and uridine), and rich in antioxidant phenolic compounds, mainly caffeic acid metabolites. Our data suggest that chia seeds may be used as ingredients for the development of functional foods and dietary supplements.

Study on the Effect of Different Iodine Intake on Hippocampal Metabolism in Offspring Rats.

Iodine is an essential trace element in the human body. Severe maternal iodine deficiency during pregnancy leads to obvious intellectual disability in the offspring. The effects of iodine deficiency on brain development have been demonstrated, but there is no clear evidence of the effects of iodine excess on brain development. To clarify the effects of iodine excess on the brain development of offspring and to provide clues to the mechanisms underlying the effects of iodine deficiency and iodine excess on the brain development of offspring. In this study, animal models with different iodine intakes were constructed using potassium iodate (KIO3). The models included four experimental groups (low-iodine group one (LI, 0μg/L iodine), low-iodine group two (LII, 5μg/L iodine), high-iodine group one (HI, 3000μg/L iodine), and high-iodine group two (HII, 10000μg/L iodine)) and one control group (NI, 100μg/L iodine). There were 20 female rats in each group, and 8 offspring were chosen from each group following birth to assess metabolic alterations. The metabolites of subsets of brain hippocampal tissue were profiled by ultra-performance liquid chromatography-linked electrospray ionization quadrupole time-of-flight mass spectrometry (UPLC-ESI-QTOFMS) and the results were subjected to multivariate data analysis. Differential substances were screened by t test (p<0.05), principal component analysis (PCA), and partial least squares analysis (PLS-DA, VIP>1). The thyroid function of the female rats in the experimental group was abnormally changed. Metabolic analysis showed that the five groups were separated which revealed significant differences in hippocampal tissue metabolism among the five groups of offspring. A total of 12 potential metabolites were identified, with the majority of them being related to amino acid and energy metabolism. These metabolites are involved in various metabolic pathways, are interrelated, and may play a function in brain development. Our study highlights changes in metabolites and metabolic pathways in the brain hippocampus of offspring rats with different iodine intakes compared to controls, revealing new insights into hippocampal metabolism in offspring rats and new relevant targets.

GC-MS/MS Profiling of Plant Metabolites.

Gas chromatography coupled to electron ionization (EI) quadrupole mass spectrometry (GC-MS) is currently one of the most developed and robust metabolomics technologies. This approach allows for simultaneous measurements of large number of chemically diverse compounds including organic acids, amino acids, sugars, sugar alcohols, aromatic amines, and fatty acids. Untargeted GC-MS profiling based on full scan data acquisition requires complicated raw data processing and sometime provides ambiguous metabolite identifications. Targeted analysis using GC-MS/MS can provide better specificity, increase sensitivity, and simplify data processing and compound identification but wider application of targeted GC-MS/MS approach in metabolomics is hampered by the lack of extensive databases of MRM transitions for non-derivatized and derivatized endogenous metabolites. The focus of this chapter is the automation of GC-MS/MS method development which makes it feasible to develop quantitative methods for several hundred metabolites and use this strategy for plant metabolomics applications.

Extending the mass-to-charge scannable range of a linear ion trap mass spectrometer through quadrupolar direct current scan

Mass scan methods utilizing quadrupolar direct current (DC) scan were described and performed with a linear ion trap (LIT) through a home-built MS apparatus designed for ion/ion reactions. Both quadrupolar DC downscan and upscan are applicable to the LIT due to the shape of the related Mathieu stability diagram. Besides, a supplementary AC signal could be added to the DC scan process for resonant ejection, which largely enhances the signal-to-noise (S/N) ratios of the spectra. Compared with conventional resonance ejection RF scans, the scannable mass-to-charge (m/z) range in quadrupolar DC scan is much wider. Several protein cations were chosen as test molecules with a m/z scannable range larger than 140,000 Th. Besides, compared with three-dimensional (3D) quadrupolar ion traps (QIT), the LIT is a more suitable mass analyzer for ions with large m/z ratios due to large ion capacity and deep pseudopotential well depths for ions inside the trap. In general, DC scan with LIT offers a new way of rapid detection of ions with extra wide m/z scannable ranges, which has potential applications in research fields such as “top-down” proteomics and native mass spectrometry in the future.

Degradation of dibutyl phthalate by Paenarthrobacter sp. Shss isolated from Saravan landfill, Hyrcanian Forests, Iran

Phthalic acid esters are predominantly used as plasticizers and are industrially produced on the million ton scale per year. They exhibit endocrine-disrupting, carcinogenic, teratogenic, and mutagenic effects on wildlife and humans. For this reason, biodegradation, the major process of phthalic acid ester elimination from the environment, is of global importance. Here, we studied bacterial phthalic acid ester degradation at Saravan landfill in Hyrcanian Forests, Iran, an active disposal site with 800 tons of solid waste input per day. A di-n-butyl phthalate degrading enrichment culture was established from which Paenarthrobacter sp. strain Shss was isolated. This strain efficiently degraded 1 g L–1 di-n-butyl phthalate within 15 h with a doubling time of 5 h. In addition, dimethyl phthalate, diethyl phthalate, mono butyl phthalate, and phthalic acid where degraded to CO2, whereas diethyl hexyl phthalate did not serve as a substrate. During the biodegradation of di-n-butyl phthalate, mono-n-butyl phthalate was identified in culture supernatants by ultra-performance liquid chromatography coupled to electrospray ionization quadrupole time-of-flight mass spectrometry. In vitro assays identified two cellular esterase activities that converted di-n-butyl phthalate to mono-n-butyl phthalate, and the latter to phthalic acid, respectively. Our findings identified Paenarthrobacter sp. Shss amongst the most efficient phthalic acid esters degrading bacteria known, that possibly plays an important role in di-n-butyl phthalate elimination at a highly phthalic acid esters contaminated landfill.

Monoclonal and Oligoclonal Anti-PF4 Antibodies Mediate VITT

Background: ChAdOx1 nCoV-19 (AstraZeneca) and Ad26.COV2.S (Janssen Johnson & Johnson) vaccines against COVID-19 have been associated with thrombotic thrombocytopenic reactions referred to as vaccine-induced immune thrombotic thrombocytopenia (VITT) characterized by the presence of platelet-activating, anti-PF4 antibodies. While VITT shares key clinical features with a similar but separate entity, Heparin-induced thrombocytopenia (HIT), there appear to be important differences: 1) VITT patients have extremely high thrombosis rates and are very strongly positive in PF4-polyanion ELISAs, and 2) Many patients with VITT frequently present with refractoriness to therapy or have disease recurrence that suggests distinct antibody characteristics due to a strong autoimmune anti-PF4 response.Aims: The goal of this study was to characterize anti-PF4 antibodies in VITT.Methods: Five VITT patients were studied, one after ChAdOx1 nCoV-19 vaccination and four after Ad26.COV2. Reactivity of VITT anti-PF4 antibodies to uncomplexed PF4, PF4-Polyvinyl sulfonate (PVS), and PF4-heparin targets was evaluated, and the platelet-activating ability of these antibodies was examined in the PF4-dependent P-selectin Expression assay (PEA). Anti-PF4 antibodies were isolated from patient blood samples using PF4-treated heparin sepharose beads, and isolated antibodies were subject to mass spectrometric evaluation (Liquid Chromatography Electrospray Ionization Quadrupole time-of-flight mass spectrometry [LC-ESI-QTOF MS]).Results: Antibodies from all VITT patients recognized both uncomplexed and complexed PF4 (Fig. 1A). Interestingly, recognition of PF4 by VITT antibodies was lower if PF4 targets were complexed with polyanions, PVS, or heparin (Fig. 1A). These results contrasted with those obtained in a “classical” HIT patient which showed reactivity to PF4/polyanion complexes, but not to uncomplexed PF4 (Fig 1A). All samples activated platelets in the PEA (data not shown). Mass spectrometric evaluation of anti-PF4 antibodies isolated from VITT patients demonstrated monoclonal anti-PF4 antibodies in three patients, and bi- and tri-clonal antibodies in one patient each (a representative monoclonal antibody anti-PF4 antibody is shown in Fig 1B). Consistent with current dogma, polyclonal anti-PF4/polyanion antibodies were seen in “classical” HIT (Fig 1C). Evaluation of anti-PF4 antibodies in spontaneous HIT, a type of autoimmune HIT seen in pro-inflammatory milieus such as orthopedic surgery and infectious prodromes also demonstrated monoclonal anti-PF4 antibodies (Fig 1D). Eluates from control heparin-sepharose beads did not reveal any immunoglobulins (data not shown).Conclusion: Although development of platelet-activating anti-PF4 antibodies and the thrombotic thrombocytopenia syndrome seen after ChAdOx1 nCoV-19 and Ad26.COV2.S vaccination resembles HIT, these findings demonstrate that clonally restricted anti-PF4 antibodies mediate VITT while polyclonal anti-PF4 antibodies mediate HIT. In addition, we noted clonally-restricted anti-PF4 antibodies in another condition that does not require proximate heparin exposure, spontaneous (“autoimmune”) HIT. In VITT, the strong immune response after vaccine administration may result in the activation of a single or few pre-existing anti-PF4 reactive clones, and development of clonally restricted anti-PF4 antibodies with a similar pathophysiology to Spontaneous HIT. It is also likely that high levels of monoclonal/oligoclonal anti-PF4 antibodies cause the severe thrombotic phenotypes seen in VITT and Spontaneous HIT. The high mortality rate and reports of disease refractoriness to therapy in VITT may warrant consideration of additional therapeutic modalities like rituximab and therapeutic plasma exchange in select cases.Figure Legends: (A): VITT (Patient 1-ChAdOx1 nCoV-19; Patients 2-5, Ad26.COV2.S) patient samples were tested in ELISA against uncomplexed PF4 (white), and PF4 in complex with polyvinyl sulfonate (light grey), or unfractionated heparin (dark gray). (B-D) Mass spectrometric evaluation of anti-PF4 antibodies isolated from VITT (B), HIT (C) and spontaneous HIT patient sera (D). “Relative Intensity” refers to abundance of the Ig light chain relative to the polyclonal background. Numbers above Ig light chain peaks depict mass/charge ratios. NC- Normal control. [Display omitted] DisclosuresMurray: Mayo Clinic: Other: Has received patents for the Mass-Fix technology which has been licensed to the Binding Site with potential royalties.. Padmanabhan: Veralox Therapeutics: Membership on an entity's Board of Directors or advisory committees.

Ion scanning or ion trapping: Why not both?

The present contribution describes analogies and differences between the quadrupolar ion trap (QIT) and the quadrupole mass analyzers, shows the potentialities of their combination in a single instrument and presents a review of applications of such a technology in different fields. The first section describes the quadrupole mass filter (QMF), outlining its principles of operation and the ion sorting procedure according to the use of oscillating electric fields inducing stable trajectories to the ions allowing them to reach the detector. Multiple quadrupole systems (normally triple quadrupoles) are then explained, showing their use in tandem mass spectrometry in space experiments (MS/MS-in-space). QIT principles of operation are then examined, pointing out that in this case the use of the same combination of oscillating electric fields takes advantage of unstable ion trajectories for their sorting. Substantially, analogies and differences between QMF and QIT come out, which consist in the fact that QMF is a scanning mass analyzer, whereas QIT is a sequential mass analyzer. In addition, the section underlines that QIT is capable to perform tandem mass spectrometry in time experiments (MS/MS-in-time). Later, the possibility to use a quadrupole as a trapping system with a prevailing dimension (linear ion trap [LIT]) is taken into consideration, and the possibility to combine both QMF and LIT in a single instrument, a QTrap mass spectrometer, is illustrated. In this frame, a lot more experiment types are possible with respect to both standalone triple quadrupoles and LIT, and they are described as well. Several combinations of these QTrap features can be used in information dependent acquisition (IDA) mode, allowing the high versatility of this instrumental configuration. The second section deals with a review of applications in different fields. These are organized by kind of QTrap and IDA features and cover different topics in biological, medical, agrochemical, nutritional and environmental fields.

Metabolic Profile of C-Prenyl Coumarins Using Mass Spectrometry-Based Metabolomics

C-prenyl coumarins (C-PYCs) are compounds with similar structures and various bioactivities, which are widely distributed in medicinal plants. Until now, the metabolic characterizations of C-PYCs and the relationship between metabolism and bioactivities remain unclear. In this study, ultra-performance chromatography electrospray ionization quadrupole time-of-flight mass spectrometry-based metabolomics (UPLC-ESI-QTOF-MS) was firstly used to determine the metabolic characterizations of three C-PYCs, including meranzin hydrate (MH), isomeranzin (ISM), and meranzin (MER). In total, 52 metabolites were identified, and all of them were found to be novel metabolites. Among these metabolites, 10 were from MH, 22 were from ISM, and 20 were from MER. The major metabolic pathways of these C-PYCs were hydroxylation, dehydrogenation, demethylation, and conjugation with cysteine, N-acetylcysteine, and glucuronide. The metabolic rate of MH was much lower than ISM and MER, which was only 27.1% in MLM and 8.7% in HLM, respectively. Additionally, recombinant cytochrome P450 (CYP) screening showed that CYP1A1, 2B6, 3A4, and 3A5 were the major metabolic enzymes involved in the formation of metabolites. Further bioactivity assays indicated that all of these three C-PYCs exhibited anti-inflammatory activity, but the effects of ISM and MER were slightly higher than MH, accompanied by a significant decrease in inflammatory cytokines transcription induced by lipopolysaccharide (LPS) in macrophages RAW 264.7. Taken together, the metabolic characterizations of the three C-PYCs suggested that the side chain of the prenyl group may impact the metabolism and biological activity of C-PYCs.

Effects of extraction parameters on lipid profiling of mosses using UPLC-ESI-QTOF-MS and multivariate data analysis.

Non-target lipid profiling by using ultra-performance liquid chromatography coupled to electrospray ionization quadrupole time-of-flight mass spectrometry (UPLC-ESI-QTOF-MS) has been used extensively in the past decades in plant studies. However, the lipidomes of bryophytes have only been scarcely studied, although they are the second largest group in plant kingdom. We evaluated the effects of different cell disruption methods (no disruption, shake, ultrasound, and bead beating), and storage conditions (air-dried, freeze-dried, and fresh frozen) of five moss species (including Racomitrium lanuginosum B and D, Philonotis fontana, Sphagnum teres, and Hylocomium splendens). The lipid profiling results of each extraction parameter were analyzed by using multivariate data analysis including unsupervised principal component analysis and supervised orthogonal projections to latent structures discriminant analysis. The results showed that extraction with bead beating resulted in the highest lipid content and the most detected features, but these were caused by the contamination from plastic tubes. Minor lipid metabolite changes were found in shaking and ultrasonication methods when compared with no disruption method. Significant amounts of phosphatidylcholine, diacylglyceryltrimethylhomoserine and their lyso lipids were observed in air-dried moss tissues, whereas diacylglycerol, triacylglycerol and ceramide were mostly exclusively detected when fresh frozen tissues were used for extraction. We concluded that lipid extraction using fresh frozen samples with ultrasound assistance provide the most original lipid composition and gave a relatively high lipid content.

Polyphenolic composition and antioxidant potential of underutilized Himalayan wild edible berries by high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry.

The Himalayan wild berries Myrica esculenta, Duchesnea indica, Prunus cerasoides, and Berberis lycium were studied for the first time for polyphenolic compositions using high-performanceliquid chromatography/electrospray ionization tandemmass spectrometry. The chemical analysis of extracts resulted in the identification of 69 polyphenols, including 11 anthocyanins, 28 phenolic acids, 19 flavanoids, 10 ellagitannins, and 1 dihydrochalcone. The caffeoylquinic acids, quercetin, kaempferol, and cyanidin are the common moieties in studied wild berries. In addition, seven phenolic compounds, that is, gallic acid, vanillic acid, caffeic acid, rutin, ferulic acid, quercetin, and myricetin were quantified. Among anthocyanins, cyanidin-3-O-glucoside was the majorly quantified anthocyanin that ranged from 2.52 to 6.18% followed by delphinidin-3-O-glucoside (0.6%-5.53%). The samples were assessed for total phenolics, total anthocyanin content, and antioxidant potential. The results showed that M. esculenta and B. lycium had the highest total phenolic and anthocyanin content and exhibited promising antioxidant activity with IC50 of 32.100 ± 0.42 and 48.44 ± 0.46μg/mL, respectively. Principal component analysis was able to clearly differentiate between four underutilized Himalayan wild edible berries. The wide variability of polyphenolic constituents supports the consumption of these berries not only to achieve the nutrient requirements among local people but also as promising sources of antioxidants for the food industry.

Development of a high-throughput method for the comprehensive lipid analysis in milk using ultra-high performance supercritical fluid chromatography combined with quadrupole time-of-flight mass spectrometry

Milk lipids are one of the most complex materials in nature and are associated with many physiological functions, hence it is important to comprehensively characterize lipids profiles to evaluate the nutritional value of milk. A quick method was developed by ultra-high performance supercritical fluid chromatography coupled to electrospray ionization quadrupole time-of-flight mass spectrometry (UHPSFC-ESI-QTOF-MS) to analyze the non-polar and polar lipids profiles of cow, goat, buffalo, human milk, and infant formulas in 7 min. All chromatographic conditions were carefully optimized and their effect on the chromatographic behavior of lipid classes and species was discussed. Under optimized conditions, 12 lipid classes (triacylglycerols, diacylglycerols, monoglyceride, fatty acids, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, phosphatidylglycerol, sphingomyelin, lyso-phosphatidylcholine, and lyso-phosphatidylethanolamine) were separated and each class was further separated in single analysis to facilitate the identification. 250 lipid species in real samples were characterized and quantified. This result demonstrates the applicability of the UHPSFC-ESI-QTOF-MS method in the high-throughput and comprehensive lipid analysis of milk, and will hopefully help to provide nutritionists with the lipid distribution in different types of milk, as well as help in the design of more suitable infant formula for babies.