High-resolution Mass Analysis Research Articles

Emerging pollutants: Risks in salmon fish migration. A review

The release of pollutants into the environment is a serious ecological problem due to the significant risks they pose both to ecosystems and to human health. The review considers the issues of detection and distribution of new pollutants, as well as sources of their entry into the environment and potential impact on ecosystems. The problem of wastewater pollution is especially urgent in the context of conservation of salmonid fish species. Pharmaceutical and veterinary drugs, components of personal hygiene products, microplastics, per- and polyfluorinated substances, brominated flame retardants, oil and a number of other toxicants enter freshwater bodies, which are especially important for the early stages of the salmon development and its following migration. The presence of pollutants in the aquatic environment depends on a source, their ability to bioaccumulate, as well as to degrade in the processes of biological and abiotic purification. High-resolution mass spectrometry in combination with gas or liquid chromatography can be recommended for detection of new pollutants. Due to high mass measurement accuracy and high specificity, this method is able to effectively identify pollutants even in the complex fish matrix. When conducting non-targeted and targeted screening with highly sensitive full scanning, it is advisable to use high-resolution mass analyzers (TOF, Q-TOF, Q-Orbitrap), which allow determining a number of new compounds. A promising direction in the complex assessment of the impact of pollutants on salmon fish species and the identification of indicators characterizing this impact is metabolomics. It is based on the study of low-molecular organic compounds — metabolites, both intermediate and final metabolic products in body fluids and tissues. Metabolomic profiling allows identifying important markers of the state of a water body under the influence of pollutants, which is necessary for the development of comprehensive regulations and guidelines for the effective handling of these pollutants.

New methyl compounds using the predicted data mining approach (PDMA), coupled with the biotransformation of Streptomyces peucetius O-methyltransferase

The structural modification of natural products using biotransformation is an effective way to produce organic compounds in a regioselective and/or stereoselective manner. O-methylation is a major modification in nature. The predicted data mining approach (PDMA) can efficiently screen out biotransformable precursor candidates to produce new (functional) compounds from thousands of derivatives. Herein, an O-methyltransferase (SpOMT2884) from Streptomyces peucetius (ATCC 27952 strain) was selected to biotransform eight precursors, screened from 4364 commercially available natural compounds via PDMA. Seven of the eight precursors (calceolarioside B, isomangiferin, mangiferin, methyl chlorogenate, plantagoside, protosappanin B, and wedelolactone) could be biotransformed from SpOMT2884. Fourteen biotransformed compounds were confirmed to be methyl products with a high-resolution mass analysis. Three methyl products from two precursors (plantagoside and protosappanin B) were selected for further production and identified using nucleic magnetic resonance spectral methods. One methyl product of protosappanin B was identified as 10-O-methyl protosappanin B (1), which gained potent anti-inflammatory activity (IC50 = 76.1 ± 4.7 μM) compared with its precursor, protosappanin B (IC50 = 157.7 ± 5.0 μM). In a case study, two methyl products of plantagoside were indeed identified as novel 4′-O-methyl plantagoside (3) and 5′-O-methyl plantagoside (4). This study demonstrates that PDMA is a good in silico approach for screening biotransformable precursor candidates for the in vitro production of new or bioactive compounds from numerous natural products.

The OrbiTOF mass analyzer: Time-of-flight analysis via an orbitrap quadro-logarithmic field with periodic drift focusing

Thermo Scientific™ Orbitrap™ analyzers represent a prominent class of high-resolution mass analyzer commonly used in life sciences, and for interrogation of complex samples. Injected ions, trapped within a quadro-logarithmic field, orbit a central electrode and oscillate up and down the axis. A new class of multi-reflection time-of-flight mass analyzer has been developed based on the Orbitrap field structure plus an additional series of periodic lenses wrapped around the central axis to constrain beam dispersion. The axial and angular velocity of the injected ions was balanced so that with each axial oscillation, the ions passed through the next lens in the series, to form a tightly folded 25-m long, 3-dimensional ion path, ending with ions striking a detector surface.Performance was interrogated via experiment and simulation. 70k resolving power was observed within the relatively compact analyzer, albeit at cost to transmission. A larger design with an integrated extraction trap and greater flight energy is discussed.

Polyprenylated acylphloroglucinols from Hypericum sampsonii with cytotoxicity against pancreatic carcinomas

Chemical investigation on the 80% EtOH extract of the air dried aerial parts of Hypericum sampsonii resulted in the isolation of two new polycyclic polyprenylated derivatives, hypersampines A and B (1 and 2). The structures of the new compounds were elucidated by spectroscopic data (NMR, IR, and UV) and high resolution mass analysis. The two isolated polyprenylated acylphloroglucinols were tested in vitro for cytotoxic activities against 6 pancreatic cell lines. As a result, compounds 1 and 2 possessed modest cytotoxic activities against all the tested tumor cell lines with IC50 values less than 40 μM.

Modern Electrospray Ionization Mass Spectrometry Techniques for the Characterization of Supramolecules and Coordination Compounds.

Mass spectrometry is routinely used for myriad applications in clinical, industrial, and research laboratories worldwide. Developments in the areas of ionization sources, high-resolution mass analyzers, tandem mass spectrometry, and ion mobility have significantly extended the repertoire of mass spectrometrists; however, for coordination compounds and supramolecules, mass spectrometry remains underexplored and arguably underappreciated. Here, the reader is guided through different tools of modern electrospray ionization mass spectrometry that are suitable for larger inorganic complexes. All steps, from sample preparation and technical details to data analysis and interpretation are discussed. The main target audience of this tutorial is synthetic chemists as well as technicians/mass spectrometrists with little experience in characterizing labile inorganic compounds.

Degradation of limonene and trans-cinnamaldehyde in soil, and detection of their metabolites by UHPLC and GC-HRMS

Two commercial biopesticides were studied to determine their persistence in two soil types, such as sandy clay loam and clay loam soils. For this purpose, an orange oil–based biopesticide was used, being limonene its main ingredient. The other biopesticide was based on cinnamon extract and trans-cinnamaldehyde as its main component. Degradation of these compounds was monitored, and transformation products or metabolites were detected. Limonene and its metabolites were analyzed by gas chromatography (GC) and trans-cinnamaldehyde by ultra-high-performance liquid chromatography (UHPLC). Both techniques were coupled to a high-resolution mass (HRMS) analyzer, such as quadrupole (Q)-Orbitrap. Limonene and trans-cinnamaldehyde were rapidly degraded as result of first-order kinetics. Possible metabolites such as thymol, cymene, isoterpinolene and cymenene for limonene, and hydroxycinnamic acid for trans-cinnamaldehyde were tentatively identified. Moreover, four other metabolites of trans-cinnamaldehyde, some of them not previously described, were also detected.

Simulation study of digital waveform-driven quadrupole mass filter operated in higher stability regions for high-resolution inductively coupled plasma mass spectrometry.

The use of a frequency-scanned digital quadrupole mass filter (QMF) with varying duty cycles shows promise for application as a high-resolution mass analyzer design for inductively coupled plasma mass spectrometry (ICP-MS). High resolution in ICP-MS is important to overcome isobaric polyatomic interferences. Here, we explore the possibility and the characteristics of using a digital quadrupole operating in higher stability regions for ICP-MS. We perform computational simulations in SIMION of a digital QMF that is operated by scanning the frequency of the digital waveform at a fixed driving voltage and various duty cycles. For ions in the atomic mass range (7-238 m/z), we investigate the expected resolution, transmission, fringe field effects, and ion trajectories. We compare different characteristics between sine and digital waveform QMF. Within the capability of current digital waveform generation technology, a digital QMF can produce variable mass resolution, from several hundred to more than 10 000. This mass resolution covers the low, medium, and high resolutions that are typical for sector-field ICP-MS. Additionally, simulations suggest that transmission of the QMF remains high at high resolution. For example, with 87.50/12.50 duty cycle (zone 4,1), resolution at 10% peak width is 10420 for m/z 80. The transmission through the quadrupole, which is constant for all isoenergetic ions, is ~2.5%, and most ion loss is due to the defocusing effects of the fringe field. Compared to sinusoidal QMFs, ions need many fewer cycles in the digital QMF to obtain high resolution. The results demonstrate that the use of a frequency-scanned, duty-cycle-modulated digital QMF as the mass analyzer for ICP-MS has the potential to produce high resolution while maintaining considerable transmission, thus overcoming most spectral interferences in elemental MS.

Synthesis and Antiproliferative Properties of Some Spirocyclic Pyrimidine Hydrazones

AbstractA new series of hydrazone derivatives incorporating spirocyclic aminopyrimidine with aryl alkanesulfonates were synthesized. The targeted products were obtained in excellent yields via an acid catalyzed reaction of spirocyclic hydrazinopyrimidine with aryl aldehyde sulfonates. The structures of the synthesized hydrazones were studied by the different spectroscopic tools, in addition to the high resolution mass analysis. The antiproliferative activities of the compounds were tested in vitro against the National Cancer Institute (NCI) 60 cell panel by the Developmental Therapeutics Program. Amongst the tested hydrazone compounds, the methansulfonate derivatives showed the highest activities against lung, brain, kidney, and breast cancers.

CSF N-Glycomics Using High-Throughput UPLC-ESI Techniques in Alzheimer's Disease.

In this chapter, we will present a high-throughput method applied in our laboratory for the structural elucidation of the cerebrospinal fluid (CSF) N-glycome. This methodology is based on a commercial equipment developed by WATERS™ to speed up N-deglycosylation and N-glycan labeling of glycoproteins of pharmaceutical and biological interest such as monoclonal antibodies. This analytical kit is sold under the trade name of RapiFluor-MS (RFMS). We have slightly modified the methodology, increasing the glycosylation time and using a high-resolution mass analyzer for the analysis of CSF N-glycans, thus obtaining a high-throughput method (up to 96 samples simultaneously), mass accuracy better than 5ppm, and the ability to separate and identify isomers.

Approximate calculation of static mass analyzers based on two-dimensional fields

The concurrent utilization of electric and magnetic fields in static mass analyzers proves highly effective in sparticle separation solely based on their masses, thereby ensuring remarkable analytical capabilities. High-resolution mass analyzers employ an achromatizing electrostatic system to precisely focus ion radiation by energy. Devices featuring zero energy dispersion are created through the collaborative action of electric achromatizing systems and the magnetic field within the analyzer, resulting in ion separation based solely on mass. Expanding the ion beam prior to its magnetic field exposure enhances the overall quality of static mass analyzers. This article delves into a static mass analyzer with dual ion beam focusing, elucidating the ion acceleration and deceleration processes at electrode boundaries, their focalization via a transaxial lens, and the impact of the electrostatic prism's refractive surface on the mass analyzer's "quality" parameter. Additionally, the transition from a sector homogeneous magnetic field to the fields generated by a two-dimensional magnetic prism and a three-electrode transaxial focusing lens is explored to augment the mass analyzer's linear dispersion. To refine ion beam focus pre-magnetic field entry, the proposed adoption of a two-dimensional electrostatic prism promises heightened linear dispersion in the mass analyzer while expanding the beam pre-magnetic field entry. This study also examines the impact of a two-dimensional electric prism on the charged particle beam, presenting an approximate expression for its angular energy dispersion. It is demonstrated that a prism-based mass analyzer integrating two-dimensional electric and magnetic prisms enables energy-focused operations alongside enhanced linear dispersion and reduced linear dimensions.

An Orbitrap/Time-of-Flight Mass Spectrometer for Photofragment Ion Imaging and High-Resolution Mass Analysis of Native Macromolecular Assemblies.

We discuss the design, development, and evaluation of an Orbitrap/time-of-flight (TOF) mass spectrometry (MS)-based instrument with integrated UV photodissociation (UVPD) and time/mass-to-charge ratio (m/z)-resolved imaging for the comprehensive study of the higher-order molecular structure of macromolecular assemblies (MMAs). A bespoke TOF analyzer has been coupled to the higher-energy collisional dissociation cell of an ultrahigh mass range hybrid quadrupole-Orbitrap MS. A 193 nm excimer laser was employed to photofragment MMA ions. A combination of microchannel plates (MCPs)-Timepix (TPX) quad and MCPs-phosphor screen-TPX3CAM assemblies have been used as axial and orthogonal imaging detectors, respectively. The instrument can operate in four different modes, where the UVPD-generated fragment ions from the native MMA ions can be measured with high-mass resolution or imaged in a mass-resolved manner to reveal the relative positions of the UVPD fragments postdissociation. This information is intended to be utilized for retrieving higher-order molecular structural details that include the conformation, subunit stoichiometry, and molecular interactions as well as to understand the dissociation dynamics of the MMAs in the gas phase.

Prediction of Fragmentation Pathway of Azilsartan Protonated Ion Using High Resolution Mass Spectrometry (HR-MS/MS) with Q-TOF Analyzer- A Perspective Study

Introduction: UPLC coupled with a high-resolution mass analyzer i.e, Q-TOF analyzer with electrospray ionization (ESI) source using collision-induced dissociation (CID) method was applied to study the characteristic product ions of azilsartan protonated ion. Method: The experimental results of high-resolution mass spectra explained the elemental compositions of product ions accurately and reasonable fragmentation pathways were proposed for azilsartan protonated ions. Calculated mass error in parts per million (ppm) for molecular ions and product ions and results of mass error found from this study from 0.2 ppm to 3.5 ppm. Result: The characteristic fragmentation pathways were helpful to analyze and interpret the stability and possible degradation pathway of the parent ion. Conclusion: The present study explains the significant role of high-resolution mass spectrometry in the structural analysis of the protonated ion of azilsartan.

Development of a high-resolution mass-spectrometry-based method and software for human leukocyte antigen typing.

The human leukocyte antigen (HLA) system plays a critical role in the human immune system and is strongly associated with immune recognition and rejection in organ transplantation. HLA typing method has been extensively studied to increase the success rates of clinical organ transplantation. However, while polymerase chain reaction sequence-based typing (PCR-SBT) remains the gold standard, cis/trans ambiguity and nucleotide sequencing signal overlay during heterozygous typing present a problem. The high cost and low processing speed of Next Generation Sequencing (NGS) also render this approach inadequate for HLA typing. To address these limitations of the current HLA typing methods, we developed a novel typing technology based on nucleic acid mass spectrometry (MS) of HLA. Our method takes advantage of the high-resolution mass analysis function of MS and HLAMSTTs (HLA MS Typing Tags, some short fragment PCR amplification target products) with precise primer combinations. We correctly typed HLA by measuring the molecular weights of HLAMSTTs with single nucleotide polymorphisms (SNPs). In addition, we developed a supporting HLA MS typing software to design PCR primers, construct the MS database, and select the best-matching HLA typing results. With this new method, we typed 16 HLA-DQA1 samples, including 6 homozygotes and 10 heterozygotes. The MS typing results were validated by PCR-SBT. The MS HLA typing method is rapid, efficient, accurate, and readily applicable to typing of homozygous and heterozygous samples.

A high resolution Fourier transform ion trap enabled by image current splicing: a theoretical study.

The relatively high work pressure within an ion trap has limited the implementation of the Fourier transform technique for high resolution mass analysis. The main reason is that high buffer gas pressure will cause the rapid decay of ion oscillations. In this study, an image current splicing method based on the filter diagonalization method (FDM) and the Hilbert transform was developed to increase the resolving power of nondestructive mass analysis in a linear ion trap. First, multiple repeated experiments (or ion trajectory simulations) were performed to collect multiple sets of data. Using the FDM, the frequency component distribution was extracted from short image current transients collected from each experiment. The Hilbert transform was then applied to calculate and normalize the decay envelope of each transient. The relative abundance was calculated by counting the envelopes. Finally, image current transients collected from these multiple experiments were spliced and merged into a whole signal with much longer duration and continuous phase. This splicing method could effectively increase the duration of the image current, and thus improve the mass resolution of the ion trap mass analyzer. The mass resolution (m/Δm) was improved from 183.5 to 5.8 × 103, and the average relative difference was 2.8%. The proposed method resolved 3 adjacent peaks which originally could not be resolved from the raw signal by the fast Fourier transform (FFT). Besides simulated data, this method was also applied to the experimental data collected from a Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometer. The influence of electronic noise on the proposed method was also discussed in this study.

Rapid analysis of urine using mass spectrometry with atmospheric pressure laser plasma ionization

Atmospheric pressure laser plasma ionization (APLPI) technique was studied in application to the problem of rapid determination of physiologically active substances in urine. The objectives of this study included investigation by APLPI method of urine samples obtained from treated patients; determination of analytical characteristics of the method; comparative studies of APLPI and HPLC-MS/MS methods in terms of signal-to-noise ratio and evaluation of the possibility of screening urine samples without sample preparation by APLPI method. The instrumental implementation of the method is based on the use of a high-resolution mass analyzer "Orbitrap", in which the standard ion source was replaced by the in-lab developed APLPI source, combined with thermal desorption sample introduction. Urine samples, in which physiologically active substances, in particular barbiturates, carbаmazepine, thiapride, sulpiride, psilocin, amphetamine, tramadol, diphenhydramine, and others were preliminarily detected by HPLC-MS/MS, were investigated. The APLPI method was found to be capable of detecting all these analytes. The duration of the analysis of one sample does not exceed one minute. Using the "dilution in matrix" method calibration curves for a number of compounds were constructed and the analytical characteristics of the method determined. The calibration curves are linear in the range of three orders of magnitude. The mean value of the relative standard deviation of the ion signal in this range was 24%. The limits of detection found for the determination of diphenhydramine, lidocaine and phenobarbital in urine extracts were 0.7 ng/ml, 0.5 ng/ml and 10 ng/ml, respectively. Comparative studies showed that APLPI and HPLC-MS/MS methods were characterized by close values of signal-to-noise ratios. The advantages of the APLPI method are in the rapidity, retrospective analysis, and small sample volume. The limitation of the method is the necessity of desorption of the determined compounds into the gas phase. The possibility of using the APLPI method for screening physiologically active substances in urine without sample preparation has been demonstrated.

Effect of Jitter on Digital Mass Filter Analysis in Higher Stability Zones

Waveform reproducibility is a critical factor for performing high resolution mass analysis with digitally operated quadrupole mass filters and traps operating in higher stability zones. In this work, Hill equation-based stability calculations were used to define the effect of period jitter on mass analysis in higher stability zones. These calculations correlate well with experimental observations in higher stability zones. Comparison of experiment to theory supplies the basis for defining jitter-based expectations and limits for mass analysis in higher zones.

Unprecedented formation of azulicorroles from the scrambling of azulitripyrrane

In this study, we have prepared new tripyrranes embedded with azulene bearing two meso-pentafluorophenyl substituents. They were obtained as diastereomers in high yield which were separated using conventional column chromatography. Their distinct solution structures were unambiguously confirmed by NMR and mass analysis. Our synthetic strategy is simple and straightforward due to the reactivity of 1,3-positions of azulene, which underwent Lewis acid-catalyzed condensation with pyrrole-2-carbinol. These diastereomers were further subjected to acid-catalyzed (CF3COOH) condensation with pyrrole dicarbinol to afford the target N-confused azuliporphyrin (reduced form) in trace amount. However, we obtained the tris-meso-pentafluorophenyl substituted azulicorrole (AzCorA) as major product due to an unprecedented scrambling of the azulitripyrranes. The structural characterization of newly synthesized key precursors and macrocycles was done with the aid of high-resolution mass analyses, and 1H and 2D NMR analyses. The single crystal X-ray structure obtained for AzCorArevealed that the azulene subunit is tilted by 34.03[Formula: see text] due to the steric congestion of inner-NHs.

Comprehensive multi-attribute method workflow for biotherapeutic characterization and current good manufacturing practices testing.

The multi-attribute method (MAM) is a liquid chromatography-mass spectrometry (LC-MS)-based method that is used to directly characterize and monitor numerous product quality attributes (PQAs) at the amino acid level of a biopharmaceutical product. MAM enables identity testing based on primary sequence verification, detection and quantitation of post-translational modifications and impurities. This ability to simultaneously and directly determine PQAs of therapeutic proteins makes MAM a more informative, streamlined and productive workflow than conventional chromatographic and electrophoretic assays. MAM relies on proteolytic digestion of the sample followed by reversed-phase chromatographic separation and high-resolution LC-MS analysis in two phases. First, a discovery study to determine quality attributes for monitoring is followed by the creation of a targeted library based on high-resolution retention time plus accurate mass analysis. The second aspect of MAM is the monitoring phase based on the target peptide library and new peak detection using differential analysis of the data to determine the presence, absence or change of any species that might affect the activity or stability of the biotherapeutic. The sample preparation process takes between 90 and 120 min, whereas the time spent on instrumental and data analyses might vary from one to several days for different sample sizes, depending on the complexity of the molecule, the number of attributes to be monitored and the information to be detailed in the final report. MAM is developed to be used throughout the product life cycle, from process development through upstream and downstream processes to quality control release or under current good manufacturing practices regulations enforced by regulatory agencies.

Improving Ion Mobility Mass Spectrometry of Proteins through Tristate Gating and Optimization of Multiplexing Parameters.

Coupling drift tube ion mobility (IM) to Fourier transform mass spectrometry (FT-MS) affords the opportunity for gas-phase separation of ions based on size and conformation with high-resolution mass analysis. However, combining IM and FT-MS is challenging because ions exit the drift tube on a much faster time scale than the rate of mass analysis. Fourier transform (FT) and Hadamard transform multiplexing methods have been implemented to overcome the duty-cycle mismatch, offering new avenues for obtaining high-resolution, high-mass-accuracy analysis of mobility-selected ions. The gating methods used to integrate the drift tube with the FT mass analyzer discriminate against the transmission of large, low-mobility ions owing to the well-known gate depletion effect. Tristate gating strategies have been shown to increase ion transmission for drift tube IM-FT-MS systems through implementation of dual ion gating, controlling the quantity and timing of ions through the drift tube to reduce losses of slow-moving ions. Here we present an optimized set of multiplexing parameters for tristate gating ion mobility of several proteins on an Orbitrap mass spectrometer and further report parameters for increased ion transmission and mobility resolution as well as decreased experimental times from 15 min down to 30 s. On average, peak intensities in the arrival time distributions (ATDs) for ubiquitin increased 2.1× on average, while those of myoglobin increased by 1.5× with a resolving power increase on average of 11%.

Advancing Orbitrap Measurements of Collision Cross Sections to Multiple Species for Broad Applications.

Measurement of collision cross section (CCS), a parameter reflecting an ion's size and shape, alongside high-resolution mass analysis extends the depth of molecular analysis by providing structural information beyond molecular mass alone. Although these measurements are most commonly undertaken using a dedicated ion mobility cell coupled to a mass spectrometer, alternative methods have emerged to extract CCSs directly by analysis of the decay rates of either time-domain transient signals or the FWHM of frequency domain peaks in FT mass analyzers. This information is also accessible from FTMS mass spectra obtained in commonly used workflows directly without the explicit access to transient or complex Fourier spectra. Previously, these experiments required isolation of individual charge states of ions prior to CCS analysis, limiting throughput. Here we advance Orbitrap CCS measurements to more users and applications by determining CCSs from commonly available mass spectra files as well as estimating CCS for multiple charge states simultaneously and showcase these methods by the measurement of CCSs of fragment ions produced from collisional activation of proteins.