Are Internal Fragments Observable in Electron Based Top-Down Mass Spectrometry?

Protein tandem mass spectrometry (MS/MS) often generates sequence-informative fragments from backbone bond cleavages near the termini. This lack of fragmentation in the protein interior is particularly apparent in native top-down mass spectrometry (MS). Improved sequence coverage, critical for reliable annotation of posttranslational modifications and sequence variants, may be obtained from internal fragments generated by multiple backbone cleavage events. However, internal fragment assignments can be error prone due to isomeric/isobaric fragments from different parts of a protein sequence. Also, internal fragment generation propensity depends on the chosen MS/MS activation strategy. Here, we examine internal fragment formation in electron capture dissociation (ECD) and electron transfer dissociation (ETD) following native and denaturing MS, as well as LC/MS of several proteins. Experiments were undertaken on multiple instruments, including quadrupole time-of-flight, Orbitrap, and high-field Fourier-transform ion cyclotron resonance (FT-ICR) across four laboratories. ECD was performed at both ultrahigh vacuum and at similar pressure to ETD conditions. Two complementary software packages were used for data analysis. When feasible, ETD-higher energy collision dissociation MS3 was performed to validate/refute potential internal fragment assignments, including differentiating MS3 fragmentation behavior of radical versus even-electron primary fragments. We show that, under typical operating conditions, internal fragments cannot be confidently assigned in ECD or ETD. On the other hand, such fragments, along with some b-type terminal fragments (not typically observed in ECD/ETD spectra) appear at atypical ECD operating conditions, suggesting they originate from a separate ion-electron activation process. Furthermore, atypical fragment ion types, e.g., x ions, are observed at such conditions as well as upon EThcD, presumably due to vibrational activation of radical z-type ions.

Electron Capture vs Transfer Dissociation for Site Determination of Tryptic Peptide Tyrosine Sulfation: Direct Detection of Fibrinogen Sulfation Sites and Identification of Novel Isobaric Interferences.

Tyrosine sulfation, an understudied but crucial post-translational modification, cannot be directly detected in conventional nanoflow liquid chromatography-tandem mass spectrometry (nanoLC-MS/MS) due to the extreme sulfate lability. Here, we report the detection of sulfate-retaining fragments from LC-electron capture dissociation (ECD) and nanoLC-electron transfer higher energy collision dissociation (EThcD). Sulfopeptide candidates were identified by Proteome Discoverer and MSFragger analysis of nanoLC-HCD MS/MS data and added to inclusion lists for LC-ECD or nanoLC-EThcD MS/MS. When this approach failed, targeted LC-ECD with fixed m/z isolation windows was performed. For the plasma protein fibrinogen, the known pyroglutamylated sulfopeptide QFPTDYDEGQDDRPK from the beta chain N-terminus was identified despite a complete lack of sulfate-containing fragment ions. The peptide QVGVEHHVEIEYD from the gamma-B chain C-terminus was also identified as sulfated or phosphorylated. This sulfopeptide is not annotated in Uniprot but was previously reported. MSFragger further identified a cysteine-containing peptide from the middle of the gamma chain as sulfated and deamidated. NanoLC-EThcD and LC-ECD MS/MS confirmed the two former sulfopeptides via sulfate-retaining fragment ions, whereas an unexpected fragmentation pattern was observed for the third sulfopeptide candidate. Manual interpretation of the LC-ECD spectrum revealed two additional isobaric identifications: a trisulfide-linked cysteinyl-glycine or a carbamidomethyl-dithiothreiotol covalent adduct. Synthesis of such adducts confirmed the latter identity.

Determination of the activity and nuclear decay data of 157Tb.

Terbium-157 was radiochemically extracted from an irradiated tantalum target. Since the resulting material contained a significant impurity of 158Tb, 157Tb was isotopically purified using laser resonance ionization at the RISIKO mass separator in Mainz and then implanted on an aluminum (Al) foil. The implanted 157Tb was measured by two different calibrated gamma-ray spectrometers to determine photon emission rates. After dissolving the Al foil, a high purity 157Tb solution was obtained. The corresponding activity concentration was determined with a low relative uncertainty of 0.52% through a combination of liquid scintillation counting using the TDCR method and 4π(X,e)(LS)-(X,γ)(CeBr3) coincidence counting. By combining the results from all measurement techniques, emission intensities for K X-rays and gamma-rays were derived and found to be 16.05(31)% and 0.0064(2)%, respectively. The probability for K electron capture of the first forbidden non-unique transition to the ground state was determined to be 17.16(35)%. The probabilities for the electron-capture branch to the excited level and the ground state were found to be 0.084(4)% and 99.916(4)%, respectively. A Q+ value of 60.23(18) keV was estimated based on simplified BetaShape calculations, assuming an allowed transition.

Streamlining the Analysis of Proteins from Snake Venom.

Investigating snake venom is necessary for developing new treatments for envenoming and harnessing the therapeutic potential that lies within venom toxins. Despite considerable efforts in previous research, several technical challenges remain for characterizing the individual components within such complex mixtures. Here, we present native and top-down mass spectrometry (MS) workflows that enable the analysis of individual venom proteins within complex mixtures and showcase the utility of these methodologies on King cobra (Ophiophagus hannah) venom. First, we coupled ion mobility spectrometry for separation and electron capture dissociation for charge reduction to resolve highly convoluted mass spectra containing multiple proteins with masses ranging from 55 to 127 kDa. Next, we performed a top-down glycomic analysis of a 25.5 kDa toxin, showing that this protein contains a fucosylated complex glycan. Finally, temperature-controlled nanoelectrospray mass spectrometry facilitated the top-down sequence analysis of a ß-cardiotoxin, which cannot be fragmented by collisional energy due to its disulfide bond pattern. The work presented here demonstrates the applicability of new and promising MS methods for snake venom analysis.

Nitrogen can substitute helium as a mobile phase in the analysis of wastewater treatment matrices for persistent organic pollutants by gas chromatography - electron capture detection system.

Municipal wastewater treatment plants are required to monitor persistent organic pollutants (POPs) in their wastewater treatment related discharges and to assess the impact of the discharges on the environment and public health. One tool for monitoring chlorinated organic pollutants particularly is a gas chromatographic (GC) system coupled to a pair of halogen-specific electron capture detectors (ECDs) with helium (He) as the mobile phase. He supplies, however, has become inconsistent and unreliable lately. In its place, N2 gas is evaluated in this study as a potential substitute for He in quantifying organochlorine pesticides, polychlorinated biphenyls, chlordane congeners and toxaphene in wastewater treatment related matrices (influent, effluent, benthic sediment, mussel tissue, and biosolids/sludge). N2 is inert, inexpensive and requires no additional hardware to incorporate into the basic functions of a GC-ECD. Our results show that, with the usual data quality controls (blank, laboratory control, matrix spike/duplicate and proficiency testing samples, and the fact that certified reference materials data met requirements), N2 can replace He for regulatory purposes. And when necessary, the N2-based retention times (tN) can be predicted reliably from He-based retention times (tHe), irrespective of column chemistry or POPs (here: tN = 1.90tHe + 0.04, R2 = 0.996).

Sequencing of Phosphopeptides Using a Sequential Charge Inversion Ion/Ion Reaction and Electron Capture Dissociation Workflow.

Protein phosphorylation, a common post-translational modification (PTM), is fundamental in a plethora of biological processes, most importantly in modulating cell signaling pathways. Matrix-assisted laser desorption/ionization (MALDI) coupled to tandem mass spectrometry (MS/MS) is an attractive method for phosphopeptide characterization due to its high speed, low limit of detection, and surface sampling capabilities. However, MALDI analysis of phosphopeptides is constrained by relatively low abundances in biological samples and poor relative ionization efficiencies in positive ion mode. Additionally, MALDI tends to produce singly charged ions, generally limiting the accessible MS/MS techniques that can be used for peptide sequencing. For example, collision induced dissociation (CID) is readily amendable to the analysis of singly charged ions, but results in facile loss of phosphoric acid, precluding the localization of the PTM. Electron-based dissociation methods (e.g., electron capture dissociation, ECD) are well suited for PTM localization, but require multiply charged peptide cations to avoid neutralization during ECD. Conversely, phosphopeptides are readily ionized using MALDI in negative ion mode. If the precursor ions are first formed in negative ion mode, a gas-phase charge inversion ion/ion reaction could then be used to transform the phosphopeptide anions produced via MALDI into multiply charged cations that are well-suited for ECD. Herein we demonstrate a multistep workflow combining a charge inversion ion/ion reaction that first transforms MALDI-generated phosphopeptide monoanions into multiply charged cations, and then subjects these multiply charged phosphopeptide cations to ECD for sequence determination and phosphate bond localization.

Occurrence of N-nitrosamines in the atmosphere and human health risk: A case study in an urban area of Chuncheon, Gangwon State, South Korea.

This study aimed to investigate the occurrence of eight nitrosamines (NAs) in particulate (PM2.5) and gaseous phases and assess the human health risk associated with these compounds in an urban area of Chuncheon, Gangwon State, South Korea, across four sampling seasons. The findings revealed that the total concentrations of eight NAs measured during the sampling period exceeded the public health recommendation of 0.3 ng/m3 provided by the Norwegian Institute of Public Health, indicating a potential human health risk from NA exposures. In particular, the average total NA concentration observed in the gaseous samples during the winter of 2021 was 18.1 ± 6.46 ng/m3. The primary emission sources could potentially impact the concentrations of NAs in the atmosphere due to their significant positive correlation with primary emission species such as NO2, CO, and SO2. Moreover, the levels of particulate NAs during the summer were negatively correlated with O3, suggesting that their formation might be influenced by ozonation in the aqueous aerosol phase. In addition, the total NA concentrations measured in the gaseous phase were four to six times higher than those measured in the PM2.5 phase throughout the sampling period. Thus, domestic sources have the potential to impact the pollution levels of the research area more significantly than long-range atmospheric transport. In particular, the highest concentrations of NAs in the gas phase were observed during the winter, while the lowest concentrations were recorded in the summer, possibly influenced by photolysis. Nevertheless, the study suggested that tertiary amines might contribute to the presence of gaseous NAs in sunlight. Consequently, further studies focusing on the occurrence of tertiary amines in the gas phase should be considered. The cumulative lifetime cancer risks estimated from inhalation exposure exceeded the acceptable risk level of 10⁻6 for all age groups across all four seasons. Therefore, it is crucial to implement effective control measures to mitigate potential health risks associated with exposure to NAs.

In situ formation of natural deep eutectic solvent on membrane after fat hydrolysis for lindane isomers determination in peanut paste.

In this work a sample pretreatment approach assumed liquid-liquid microextraction based on the in situ formation of a hydrophobic natural deep eutectic solvent on a hydrophobic membrane impregnated with natural terpenoid was developed. The procedure included alkaline hydrolysis of a food sample containing fat to form fatty acids, which acted as precursors for the in situ formation of the deep eutectic solvent with natural terpenoid. Two processes were observed on the membrane surface: in situ formation of the hydrophobic deep eutectic solvent and liquid-liquid microextraction of the target analytes. After microextraction, the membrane containing the analytes was easily removed from the sample solution. The developed approach was applied to the separation and preconcentration of hydrophobic organochlorine pesticides (ɑ-hexachlorocyclohexane and γ-hexachlorocyclohexane) from a hydrophobic sample matrix (peanut paste), followed by their determination by gas chromatography with electron capture detection. Under optimal conditions, the limits of detection and quantification for both analytes were 0.3 and 1.0 µg kg-1, respectively. The procedure allowed the separation of fat-soluble analytes from a complex sample matrix with a high content of fat. The extraction recoveries were in the range of 93-95 %.

Post-synthetic modification of covalent organic framework for efficient adsorption of organochlorine pesticides from cattle's milk.

Analysis of organochlorine pesticides (OCPs) residues in milk faces a significant challenge. Herein, a sea urchin structured covalent organic framework bearing boric acid groups named COF-B(OH)2 was synthesized and applied as a coating material for solid-phase microextraction (SPME) of the OCPs in cattle's milk. Its performance was superior to that of three commonly used commercial SPME fibers, which could be due to the coexistence of hydrogen bonding, halogen bonding, π-stacking and electrostatic interactions. Besides, the fiber coating displayed good stability and reusability. After optimization, a COF-B(OH)2 based SPME coupled with gas chromatography-electron capture detection was established for the sensitive detection of the OCPs from milk samples. The limits of detection (S/N = 3) were between 0.04 and 1.00 µg kg-1. Satisfactory accuracy was achieved with the method recoveries in the range of 87.5 % to 112.5 %. These results manifest the feasibility of the COF-B(OH)2 coated fiber for the enrichment of the trace OCPs from milk samples.

Preparation of a magnetic molecularly imprinted polymer on fibrous silica nanosphere via self-polycondensation for micro solid-phase extraction of chlorpyrifos.

Throughout this research, a new magnetic molecularly imprinted polymer on fibrous silica nanosphere was prepared through self-polycondensation. The selective extraction of chlorpyrifos was performed by the synthesized sorbent and as a determination system, a gas chromatography-electron capture was applied. The formation of sorbent was confirmed through the use of Fourier transform infrared spectroscopy and field emission scanning electron microscopy techniques. The parameters affecting the extraction efficacy of the proposed method were scrutinized in an optimized way. The linear range and the detection limit of the studied method were 0.003-0.3 and 0.001 ng mL-1, respectively. The relative standard deviations were 4.1-5.2 and 5.6-7.6 % for intra- and inter-day (n = 3), respectively. To assess the performance of the proposed method, some water and fruit samples were analyzed and the extraction recoveries of 83-109 % were obtained. These results revealed the method's performance in the analysis of chlorpyrifos in real samples.

Liquid scintillation efficiencies, gamma-ray emission intensities, and half-life for Gd-153.

Gadolinium-153 was standardized for activity by live-timed anticoincidence counting and an ampoule was submitted to the international reference system (SIR). Absolute emission intensities for the main γ rays were determined with calibrated high-purity germanium (HPGe) and lithium-drifted silicon (Si(Li)) detectors. A revised decay scheme is indicated, with no probability of direct electron capture to the 153Eu ground state. Triple-to-double coincidence ratio (TDCR) efficiency curves indicate that the revised decay scheme is consistent with experiment. Half-life measurements agree with a previous NIST determination and show no sensitivity to chemical environment.

Non-radioactive electron capture detector based on soft X-ray ionization for gas chromatography - A possible replacement for radioactive detectors.

In this paper, we present a new electron capture detector based on a compact X-ray tube (X-ECD) for electron generation by soft X-ray radiation instead of using a radioactive source. ECDs are commonly used in many laboratories as standard GC detectors since their invention in the 1950s, especially for highly sensitive detection of halogenated substances, pesticides or other environmental pollutants. However, due to unsatisfactory alternatives, many ECDs are still used with radioactive ß-emitters, which is difficult and expensive in most applications today due to legal restrictions. The new X-ECD contains a small X-ray tube for generating free electrons by ionizing the carrier gas like in radioactive ECDs. Thus, no additional dopants or special gases are required. The X-ECD has limits of detection in the pptv range and shows linearity over a wide concentration range. Furthermore, the used X-ray tube shows good long-term stability. So far, we have operated the X-ray tube continuously for about one year without notable degradation. However, in case of future degradation, the X-ECD can still be operated with the same sensitivity by simple adjusting the set point current in constant current mode. This makes calibration robust against possible degradation of the X-ray tube. In combination with a conventional gas chromatograph, the X-ECD is able to detect halogenated hydrocarbons and even low volatile pesticides without any peak distortion such as tailing. Thereby a minimum detectability in the upper fg/µL range for Lindane was reached, which is similar when compared to radioactive ECDs.

Trihalomethanes and physicochemical quality of drinking water in Addis Ababa, Ethiopia.

Background: Trihalomethanes (THMs) are the most dominant fraction of all the byproducts formed during chlorination of water. Disinfection by product (DBP) formation in water is a function of numerous factors, including pH, temperature, residual chlorine, source water characteristics, and organic matter. No study has determined the THM level in the drinking water supply of Addis Ababa, Ethiopia. Methods: A cross-sectional design was conducted to collect water samples in the water supply distribution networks of Addis Ababa, Ethiopia. Twenty-one (21) sampling stations yielded a total of one hundred twenty (120) samples of drinking water. The sample handling and collection procedures were carried out in accordance with USEPA guidelines. A DB-5 capillary column was used to separate the THMs, which were detected using GC-ECD (gas chromatography-electron capture detector). Spectrophotometric and in situ methods were used for physicochemical parameters. Redundancy analysis (RDA) was used for data analysis of trihalomethanes and environmental variables using CANOCO 4.5. Results: The mean concentration of total trihalomethanes in drinking water in Addis Ababa was 76.3 µg/L. The concentration of chloroform in the drinking water supply in Addis Ababa, Ethiopia, ranged between 4.03 and 79.4 µg/L. The mean total THMs in the Legedadi and Gefersa water supply systems were 77.4 µg/L and 69.66 µg/L, respectively. The residual chlorine, phosphates, UV absorbance at 254 nm, and combined chlorine had positive correlations with THM formation. However, electron conductivity had a negative correlation with THM formation. Conclusions: Chloroform contributed the most to TTHMs in nearly all samples. The residual chlorine, UV absorbance, phosphate and hardness as calcium, and electron conductivity were found to be the main predictors determining the abundance and distribution of trihalomethanes. The monitoring and regulation of the THMs is required on a regular basis to analyse trends and guide the water treatment and distribution system.

[Determination of short- and medium-chain chlorinated paraffins in different components of human blood using gas chromatography-electron capture negative ion-low resolution mass spectrometry].

Short- and medium-chain chlorinated paraffins (SCCPs and MCCPs) have attracted significant attention because of their persistence, biotoxicity, bioaccumulation, and long-range migration. Given their worldwide detection in a variety of environmental matrices, concerns related to the high exposure risks of SCCPs and MCCPs to humans have grown. Thus, knowledge of the contamination patterns of SCCPs and MCCPs and their distribution characteristics in the vivo exposure of humans is of great importance. However, little information is available on the contamination of SCCPs and MCCPs in human blood/plasma/serum, mainly because of the difficulty of sample preparation and quantitative analysis. In this study, a new blood sample pretreatment method based on Percoll discontinuous density gradient centrifugation was developed to separate plasma, red blood cells, white blood cells, and platelets from human whole blood. A series of Percoll sodium chloride buffer solutions with mass concentrations of 1.095, 1.077, and 1.060 g/mL were placed in a centrifuge tube from top to bottom to establish discontinuous density gradients. The dosage for each density gradient was 1.5 mL. Human whole blood samples mixed with 0.85% sodium chloride aqueous solution were then added to the top layer of the Percoll sodium chloride solution. After centrifugation, the whole blood was separated into four components. The plasma was located at the top layer of the centrifuge tube, whereas the platelets, white blood cells, and red blood cells were retained at the junction of the various Percoll sodium chloride solutions. The sampling volume of human whole blood and incubation time were optimized, and results indicated that an excessively long incubation time could lead to hemolysis, resulting in a decrease in the recoveries of SCCPs and MCCPs. Therefore, a sampling volume of 1.5 mL and incubation time of 10 min at 4 ℃ were adopted. The cells of the blood components were further broken and extracted by ultrasonic pretreatment, followed by multilayer silica gel column chromatography for lipid removal. The use of 80 mL of n-hexane-dichloromethane (1∶1, v/v) and 50 mL of dichloromethane as the elution solvents (collected together) for the gel column separated the SCCPs and MCCPs from the lipid molecules in the blood samples. Gas chromatography-electron capture negative ion-low resolution mass spectrometry (GC-ECNI-LRMS) was used to determine the SCCPs and MCCPs. Quantification using the corrected total response factor with degrees of chlorination was achieved with linear corrections (R2=0.912 and 0.929 for the SCCPs and MCCPs, respectively). The method detection limits (MDLs) for the SCCPs and MCCPs were 1.57 and 8.29 ng/g wet weight (ww, n=7), respectively. The extraction internal standard recoveries were 67.0%-126.6% for the SCCPs and 69.5%-120.5% for the MCCPs. The developed method was applied to determine SCCPs and MCCPs in actual human whole blood samples. The contents of SCCPs and MCCPs were 10.81-65.23 and 31.82-105.65 ng/g (ww), respectively. Red blood cells exhibited the highest contents of CPs, followed by plasma, white blood cells, and platelets. The proportions of SCCPs and MCCPs in red blood cells and plasma were 70% and 66%, respectively. In all four components, the MCCP contents were higher than the SCCP contents, and the ratios of MCCPs to SCCPs ranged from 1.04 to 3.78. Similar congener patterns of SCCPs and MCCPs were found in the four components of human whole blood. C10-CPs and C14-CPs were predominantly observed in the SCCPs and MCCPs, respectively. In summary, a simple and efficient method was proposed to determine low concentrations of SCCPs and MCCPs in human blood with high sensitivity and selectivity. This method can meet requirements for the quantitative analysis of SCCPs and MCCPs in human blood components, thereby providing technical support for human health risk assessment.

Digital Quadrupole Isolation and Electron Capture Dissociation on an Extended Mass Range Q-TOF Provides Sequence and Structure Information on Proteins and Protein Complexes.

Electron capture dissociation (ECD) is now a well-established method for sequencing peptides and performing top-down analysis on proteins of less than 30 kDa, and there is growing interest in using this approach for studies of larger proteins and protein complexes. Although much progress on ECD has been made over the past few decades, establishing methods for obtaining informative spectra still poses a significant challenge. Here we describe how digital quadrupole (DigiQ) ion isolation can be used for the mass selection of single charge states of proteins and protein complexes prior to undergoing ECD and/or charge reduction. First, we demonstrate that the DigiQ can isolate single charge states of monomeric proteins such as ubiquitin (8.6 kDa) and charge states of large protein complexes such as pyruvate kinase (234 kDa) using a hybrid quadrupole-TOF-MS (Agilent extended m/z range 6545XT). Next, we demonstrate that fragment ions resulting from ECD can be utilized to provide information about the sequence and structure of the cytochrome c/heme complex and the ubiquitin monomer. Lastly, an especially interesting result for DigiQ isolation and electron capture (EC) was noted; namely, the 16+ charge state of the streptavidin/biotin complex reveals different electron capture patterns for the biotinylated proteoforms of streptavidin. This result is consistent with previous reports that apo streptavidin exists in multiple conformations and that biotin binding shifts the conformational dynamics of the complex (Quintyn, R. Chem. Biol. 2015, 22 (55), 583-592).

Development and validation of gas chromatography with electron capture detection method using QuEChERS for pesticide residue determination in cucumber.

A modified quick, easy, cheap, efficient, rugged, and safe (QuEChERS) method coupled to gas chromatography with electron capture detection was developed for the simultaneous determination of selected electronegative pesticides, namely, chlorpyrifos-methyl (1), chlorpyrifos (2), quinolphos (3), profenofos (4), myclobutanil (5), ethion (6), fenpropathrin (7), and cypermethrin (8), in vegetables with high water content. The selected compounds and some of their metabolites have even been found in human body fluids. In addition, some of them are known or suspected carcinogens according to the World Health Organization. Extraction and cleanup parameters were optimized; thus, the original QuEChERS method was modified to minimize solvent usage by making the study eco-friendly. The developed method was validated for selectivity, specificity, linearity, precision, and accuracy using SANTE guidelines. Calibration curves showed good linearity (r > 0.99) within the test range. Precision was evaluated by intra- and inter-day experiments with an acceptable range of less than 20.0% of relative standard deviation. Recovery was evaluated at limit of quantification and was found to be in the range of 70-120%, with relative standard deviations lower than 4.21%. The proposed method is applicable for detection and monitoring of selected pesticides in one run not only in fruits and vegetables with high water content but also in samples containing large quantities of pigments/dyes.

Online protein unfolding characterized by ion mobility electron capture dissociation mass spectrometry: cytochrome C from neutral and acidic solutions.

Electrospray ionization mass spectrometry (ESI-MS) experiments, including ion mobility spectrometry mass spectrometry (ESI-IMS-MS) and electron capture dissociation (ECD) of proteins ionized from aqueous solutions, have been used for the study of solution-like structures of intact proteins. By mixing aqueous proteins with denaturants online before ESI, the amount of protein unfolding can be precisely controlled and rapidly analyzed, permitting the characterization of protein folding intermediates in protein folding pathways. Herein, we mixed various pH solutions online with aqueous cytochrome C for unfolding and characterizing its unfolding intermediates with ESI-MS charge state distribution measurements, IMS, and ECD. The presence of folding intermediates and unfolded cytochrome c structures were detected from changes in charge states, arrival time distributions (ATDs), and ECD. We also compared structures from nondenaturing and denaturing solution mixtures measured under "gentle" (i.e., low energy) ion transmission conditions with structures measured under "harsh" (i.e., higher energy) transmission. This work confirms that when using "gentle" instrument conditions, the gas-phase cytochrome c ions reflect attributes of the various solution-phase structures. However, "harsh" conditions that maximize ion transmission produce extended structures that no longer correlate with changes in solution structure.

Localization of Multiple O-Linked Glycans Exhibited in Isomeric Glycopeptides by Hot Electron Capture Dissociation.

We describe a method to obtain a comprehensive profile of multiple glycosylations in glycopeptide isoforms. We detected a wide range of abundances of various O-glycoforms in isomeric glycopeptides using hot electron capture dissociation (hot ECD) in liquid chromatography-tandem mass spectrometry. To capture low abundant glycosylated species, a prototype of a ZenoTOF 7600 system incorporating an efficient electron-activated dissociation device to perform hot ECD was operated in targeted or scheduled high-resolution multiple reaction monitoring workflows. In addition, Zeno trap pulsing was activated to enhance the sensitivity of the time-of-flight mass spectrometer. Sixty-nine O-glycopeptides of the long O-glycopeptides in tryptic bovine fetuin digest were obtained with a relative abundance range from 100 to 0.2%, which included sialylated glycans with Neu5Ac and Neu5Gc.

Efficient electro-catalyzed PMS activation on a Fe-ZIF-8 based BTNAs/Ti anode: An in-depth investigation on anodic catalytic behavior.

Phenanthrene (PHE), mainly released from coal tar and petroleum distillation, is an important kind of prevalent polycyclic aromatic hydrocarbons (PAHs) contamination in China (up to 2.38 ± 0.02 mg/kg in soil and 8668 ng/L in surface water) and other countries in the world. Metal-organic frameworks (MOFs) show promising application prospects in the decontamination field, however, suffering from the intrinsic fragility and fine powder forms. Therefore, macroscopic MOFs architecture-sandwich-like Fe-ZIF-8/blue TiO2 nanotube arrays (BTNAs)/Ti substrate (FBTT) anode with strong interfacial bonding (Fe-O-Ti and Fe-2-MIM-Ti coordination) was constructed using innovative in situ growth, condensation-crystallization-deposition, and pyrolysis methods, aiming at exploring the feasibility of MOFs-based anode/peroxymonosulfate (PMS) mediated PHE elimination, revealing the in-depth mechanisms, simultaneously overcoming the intrinsic drawbacks of MOFs. The FBTT-4 (doping content of 30 %) efficiently degraded PHE by 90.01 % and 74.5 % within 10 min at 350 µg/L and 3 mg/L, respectively, mediated by the ·OH compared to the SO4·-, 1O2, and O2·-. Post-optimized range of anodic potential enabled (i) anodic oxidation, (ii) activation of water and PMS molecules to produce active species, (iii) capture of electrons in reactants to reduce Fe3+/Ti4+ to Fe2+/Ti3+, maintaining the proportion of Fe/Ti with low valence and thus stable PMS activation capacity, and (iv) regulation of the Fe/Ti d-band center to modulate the anode adsorption capacity. The further increment in anodic potential could promote "dark photocatalysis" with a Z-scheme-like mechanism. Thus, it is proposed that the development of macroscopic MOFs-based anode, especially those with small band gaps, represents vast potentials in electrocatalytic contamination elimination. Simultaneously, the MOFs-based anode is expected to fully exploit their catalytic capacities and solve their intrinsic defects as well.

Multimodal Tandem Mass Spectrometry Techniques for the Analysis of Phosphopeptides.

Collisionally activated dissociation (CAD), infrared multiphoton dissociation (IRMPD), ultraviolet photodissociation (UVPD), electron capture dissociation and electron detachment dissociation (EDD) experiments were conducted on a set of phosphopeptides, in a Fourier transform ion cyclotron resonance mass spectrometer. The fragmentation patterns were compared and varied according to the fragmentation mechanisms and the composition of the peptides. CAD and IRMPD produced similar fragmentation profiles of the phosphopeptides, while UVPD produced a large number of complementary fragments. Electron-based dissociation techniques displayed lower fragmentation efficiencies, despite retaining the labile phosphate group, and drastically different fragmentation profiles. EDD produced complex spectra whose interpretation proved challenging.