Transform-ion Cyclotron Resonance Mass Spectrometry Research Articles

Molecular–level insights into the synergistic activation of peracetic acid by ultraviolet and ferrous ions for the degradation of sedimentation sludge water in drinking water treatment plants based on Fourier transform-ion cyclotron resonance mass spectrometry

Sedimentation sludge water (SSW) from sedimentation tanks in drinking water treatment plants contains significant amounts of dissolved organic matter (DOM) and precursors of disinfection byproducts, raising increasing concerns about the safety of finished water following SSW recycling. This study explores the alterations in bulk properties and molecular characteristics of DOM in SSW treated with ultraviolet (UV)/ferrous ions (Fe2+) and peracetic acid (PAA) using fluorescence spectroscopy and Fourier transform ion cyclotron resonance mass spectrometry (FT–ICR MS). The primary reactive species, hydroxyl radicals (·OH) and acylperoxy radicals (CH3C(O)OO·) achieved degradation rates of up to 82.0%, eliminating 84.5% of total fluorescent components and reducing molecular intensity and apparent molecular count by 90.9% and 69.8%, respectively. This treatment significantly reduced the aromaticity of DOM, leading to the formation of more oxidized and stable compounds. Reaction mechanisms indicate that ·OH and CH3C(O)OO· primarily facilitate oxygen addition and dehydrogenation, while CH3C(O)OO· promotes decarboxylation through weak electron transfer processes. The treatment preferentially removes complex DOM containing nitrogen and sulfur heteroatoms, which are key precursors of disinfection byproducts. These findings enhance our understanding of DOM transformations during UV/Fe2+/PAA treatment and provide valuable insights into the safe recycling of SSW.

Recovering Valuable Chemicals from Polypropylene Waste via a Mild Catalyst-Free Hydrothermal Process.

Waste polypropylene (PP) presents a significant environmental challenge, owing to its refractory nature and inert C-C backbone. In this study, we introduce a practical chemical recovery strategy from PP waste using a mild catalyst-free hydrothermal treatment (HT). The treatment converts 64.1% of the processed PP into dissolved organic products within 2 h in an air atmosphere at 160 °C. Higher temperatures increase the PP conversion efficiency. Distinct electron absorption and emission characteristics of the products are identified by spectral analysis. Fourier transform-ion cyclotron resonance-mass spectrometry (FT-ICR-MS) reveals the oxidative cracking of PP into shorter-chain homologues (10-50 carbon atoms) containing carboxylic and carbonyl groups. Density functional theory (DFT) calculations support a reaction pathway involving thermal C-H oxidation at the tertiary carbon sites in the polymer chain. The addition of 1% H2O2 further enhances the oxidation reaction to produce valuable short-chain acetic acids, enabling gram-scale recycling of both pure PP and disposable surgical masks from the real world. Techno-economic analysis (TEA) and environmental life cycle costing (E-LCC) analysis suggest that this hydrothermal oxidation recovery technology is financially viable, which shows significant potential in tackling the ongoing plastic pollution crisis and advancing plastic treatment methodologies toward a circular economy paradigm.

Characterization of the Bioavailability of Per- and Polyfluoroalkyl Substances in Farmland Soils and the Factors Impacting Their Translocation to Edible Plant Tissues.

In this study, various crops and farmland soils were collected from the Fen-Wei Plain, China, to investigate the bioavailability of perfluoroalkyl substances (PFAS), their accumulation in edible plant tissues, and the factors impacting their accumulation. PFAS were frequently detected in all of the crops, with total concentrations ranging from 0.61 to 35.8 ng/g. The results of sequential extractions with water, basic methanol, and acidic methanol indicate that water extraction enables to characterize the bioavailability of PFAS in soil to edible plant tissues more accurately, especially for the shorter-chain homologues. The bioavailability of PFAS was remarkably enhanced in the rhizosphere (RS) soil, with the strongest effect observed for leafy vegetables. The water-extracted Σ16PFAS in RS soil was strongly correlated with the content of dissolved organic carbon in the soil. Tannins and lignin, identified as the main components of plant root exudates by Fourier transform-ion cyclotron resonance mass spectrometry, were found to enhance the bioavailability of PFAS significantly. Redundancy analysis provided strong evidence that the lipid and protein contents in edible plant tissues play important roles in the accumulation of short- and long-chain PFAS, respectively. Additionally, the high water demand of these tissues during the growth stage greatly facilitated the translocation of PFAS, particularly for the short-chain homologues and perfluorooctanoic acid.

Comparison of dissolved organic matter composition from various sorbents using ultra-high resolution mass spectrometry

Solid phase extraction (SPE) of a variety of diverse dissolved organic matter (DOM) endmembers through eight commercially available sorbents was examined (ENV, PLEXA, PPL, HLB, Isolute 101, C18/ENV+, C18, EnvirElut) representing styrene divinylbenzene polymer (SDVB) and silica-based sorbents. We assessed dissolved organic carbon (DOC) recovery and DOM composition via 21T Fourier transform-ion cyclotron resonance mass spectrometry (FT-ICR MS). DOC recoveries and SPE-DOM composition differed more by endmember type than by sorbent. Silica-based sorbents retained DOM with many N-containing formulae, while SDVB-based sorbents retained DOM with more S-containing formulae. Extraction pH exerted a greater influence on DOM composition, notably through the presence of strong groupings composed of saturated and lowly oxygenated formulae at basic pH, and of aromatic and highly oxygenated formulae at pH 2, irrespective of endmember or sorbent. There was above 25% DOC recovery, regardless of sorbent or endmember; >90% of the relative abundance (RA) of molecular formulae were shared with PPL, which is currently the most commonly utilized sorbent for DOM. This clearly highlights the ability of the selected sorbents to retain representative DOM across diverse endmembers. Such findings may be useful for future targeted DOM studies (e.g., bioincubations, wastewater and drinking water applications) interested in focusing on specific compositional changes and will provide a better understanding of how organic carbon cycling is impacted by anthropogenic processes.

Exploring transformation of dissolved organic matters and dissolved organic nitrogen in full-scale anammox wastewater treatment: Temperature and microbial roles

Variation of dissolved organic matters (DOM) in mainstream anammox process has received limited attention. This study systematically characterized DOM and dissolved organic nitrogen (DON) in a full-scale mainstream anammox wastewater treatment plant (WWTP) using spectroscopy and Fourier transform-ion cyclotron resonance mass spectrometry. Roles of bacterial community structures related with temperatures on DOM and DON transformations were analyzed. Results indicated that the WWTP removed highly bioavailable, S-containing DOM while producing more unsaturated, aromatic, and N-containing DOM. Higher relative abundances of Proteobacteria and Chloroflexi at low temperature resulted in greater removal rates of proteins, SMP-like and humic acid-like substances. At high temperature, higher relative abundance of Actinobacteriota increased lignin production. Principal component analysis revealed that temperature significantly impacted DOM characteristics compared to DON. These findings are crucial for understanding DOM and DON transformation during mainstream anammox WWTP.

Size-resolved molecular characterization of water-soluble organic matter in atmospheric particulate matter from northern China

Atmospheric particulate matter (PM) affects visibility, climate, biogeochemical cycles and human health. Water-soluble organic matter (WSOM) is an important component of PM. In this study, PM samples with size-resolved measurements at aerodynamic cut-point diameters (Dp) of 0.01–18 μm were collected in the rural area of Baoding and the urban area of Dalian, Northern China. Non-targeted analysis was adopted for the characterization of the molecule constitutes of WSOM in different sized particles using Fourier transform-ion cyclotron resonance mass spectrometry. Regardless of the location, the composition of WSOM in Aitken mode particles (aerodynamic diameter <0.05 μm) was similar. The WSOM in accumulation mode particles (0.05–2 μm) in Baoding was predominantly composed of CHO compounds (84.9%), which were mainly recognized as lignins and lipids species. However, S-containing compounds (64.2%), especially protein and carbohydrates species, accounted for most of the WSOM in the accumulation mode particles in Dalian. The CHO compounds (67.6%–79.7%) contributed the most to the WSOM in coarse mode particles (>2 μm) from both sites. Potential sources analysis indicated the WSOM in Baoding were mainly derived from biomass burning and oxidation reactions, while the WSOM in Dalian arose from coal combustion, oxidation reactions, and regional transport.

New piperazine and morpholine derivatives: Mass spectrometry characterization and evaluation of their antimicrobial activity

Recently, pharmaceutical research has been focused on the design of new antibacterial drugs with higher selectivity towards several strains. Major issues concern the possibility to obtain compounds with fewer side effects, at the same time effectively overcoming the problem of antimicrobial resistance. Several solutions include the synthesis of new pharmacophores starting from piperazine or morpholine core units. Mass spectrometry-based techniques offer important support for the structural characterization of newly synthesized compounds to design safer and more effective drugs for various medical conditions. Here, two new piperazine derivatives and four new morpholine derivatives were synthesized and structurally characterized through a combined approach of Fourier transform-ion cyclotron resonance (FT-ICR) and Linear Trap Quadrupole (LTQ) mass spectrometry. The support of both high-resolution and low-resolution mass spectrometric data namely accurate mass measurements, isotopic distribution and MSn spectra, was crucial to confirm the success of the synthesis. These compounds were further evaluated for inhibitory activity against a total of twenty-nine Gram-positive and Gram-negative bacteria to determine the action spectrum and the antimicrobial effectiveness. Results demonstrated compounds’ antimicrobial activity against many tested bacterial species, providing an inhibitory effect linked to different chemical structure and suggesting that the new-synthesized derivatives could be considered as promising antimicrobial agents.

The dissolved organic matter from the co-decomposition of Chinese milk vetch and rice straw induces the strengthening of Cd remediation by Fe-modified biochar

Fe-modified biochar (FB) and co-using Chinese milk vetch and rice straw (MR) are two effective ways for mitigating the cadmium (Cd) contamination in paddy fields in southern China. Nevertheless, the effects of FB combined with MR on Cd passivation mechanism remain unclear. In the current study, the strengthening effects of FB induced by MR were found and the mechanisms of the extracted dissolved organic matter (DOM) from the co-decomposition of MR on Cd alleviation were investigated through pot experiment and adsorption experiment. Pot experiment demonstrated that co-incorporating FB and MR decreased available Cd by 23.1% and increased iron plaque concentration by 11.8%, resulting in a 34.7% reduction in Cd concentrations in brown rice compared with addition of FB. Furthermore, co-using FB and MR improved available nutrients in the soil. The molecular characteristics of DOM derived from the decomposition of MR (DOM-MR) were analyzed by fluorescence excitation emission matrix spectroscopy-parallel factor analysis (EEM-PARAFAC) and Fourier transform-ion cyclotron resonance mass spectrometry (FT-ICR MS). Results showed that lignin/carboxylic-rich alicyclic molecules and protein/amino sugar were the main compounds, potentially involved in the Cd binding. Adsorption experiments revealed that the addition of DOM-MR improved the functional groups, specific surface area, and negative charges of FB, inducing the strengthening of both physisorption and chemisorption of Cd(II). The maximum adsorption capacity of Fe-modified biochar after adding DOM-MR was 634 mg g−1, 1.30 times that without the addition of DOM-MR. This study suggested that co-incorporating MR, and FB could serve as an innovative practice for simultaneous Cd remediation and soil fertilization in Cd-polluted paddy fields. It also provided valuable insights and basis that DOM-MR could optimize the performances of Fe-modified biochar and enhance its potential for Cd immobilization.Graphical

Molecular transformation of petroleum compounds by hydroxyl radicals produced upon oxidation of reduced nontronite

At redox interfaces, the interaction between oxygen and structural Fe(II) in minerals produces hydroxyl radicals (●OH); however, the specific reactivity and mechanisms of oxidation of complex petroleum compounds by ●OH remain poorly understood. This study investigates the interaction mechanisms between ●OH generated from reduced nontronite, an Fe-rich clay mineral (NAu-2), and petroleum compounds by employing a suite of wet chemistry, gas chromatography mass spectrometry and Fourier transform-ion cyclotron resonance-mass spectrometry, as well as theoretical calculations. The results showed that the intrinsic structure, composition, solubility, and adsorption of petroleum molecules onto reduced NAu-2 significantly influenced their reactivity with ●OH. Specifically, the aromatic fraction of a crude oil sample exerted a minimal influence on the oxidation rate of structural Fe(II) in reduced NAu-2, but enhanced the production of ●OH by generating environmentally persistent free radicals. ●OH reacted more with water-soluble dissolved organic matter from oil compared to polar compounds. The newly generated molecules by ●OH exhibited a preference for the insoluble (polar) phase. The oxidation of petroleum hydrocarbons by ●OH was mediated through hydrogen atom abstraction and radical adduct formation, followed by dehydrogenation, hydroxylation, and carboxylation reactions. Consequently, the abundance of oxygen-containing compounds in polar compounds increased. However, specific oxidation pathways differed depending on the nature of petroleum compounds. Our results provide insights that oxidation of petroleum compounds by ●OH is vital for mitigating the negative impacts from hydrocarbon contamination and for evaluating the role of these processes in the carbon cycle in redox-active environments.

Inhibitory effect of microplastics derived organic matters on humification reaction of organics in sewage sludge under alkali-hydrothermal treatment

Alkali-hydrothermal treatment (AHT) of sewage sludge is often used to recover value-added dissolved organic matters (DOM) enriched with artificial humic acids (HA). Microplastics (MPs), as emerging contaminants in sewage sludge, can leach organic compounds (MP-DOM) during AHT, which potentially impact the characteristics of thermally treated sludge's DOM. This study employed spectroscopy and Fourier transform-ion cyclotron resonance mass spectrometry (FT-ICR-MS) to explore the impacts of MPs on DOM composition and transformation during AHT. The biological effects of DOM were also investigated by hydroponic experiments. The results showed that the leaching of MP-DOM led to a substantial increase in DOC content of DOM of thermally treated sludge. Conversely, the HA content significantly decreased in the presence of MPs, resulting in a decline of plant growth facilitation degree. FT-ICR-MS analysis revealed that the reduction in HA content was characterized by a notable decline in the abundance of O6–7 and N1–3O6–7 molecules. Reactomics results indicated that the leaching of MP-DOM inhibited the Maillard reaction but bolstered oxidation reactions. The inhibition of Maillard reaction, resulting in a decrease in crucial precursors (dicarbonyl compounds, ketoses, and deoxyglucosone), was responsible for the decrease of HA content. The primary mechanism responsible for inhibiting the Maillard reaction was the consumption of reactive amino reactants through two pathways. Firstly, the leaching of organic acids in MP-DOM caused decrease of sludge pH, leading to the protonation of amino groups. Secondly, the lipid-like compounds in MP-DOM underwent oxidation (-2H+O), producing fatty aldehydes that consumed the reactive amino reactants. These discoveries offer enhanced insights into the specific contribution of MPs to the composition, transformation, bioactivity of DOM during AHT process.

Molecular-level insights into the temperature-dependent formation dynamics and mechanism of water-soluble dissolved organic carbon derived from biomass pyrolysis smoke

Water soluble organic carbon (WSOC) derived from biomass pyrolytic smoke is deposited through atmospheric aerosols, negatively affecting aquatic ecological quality and safety. However, the temperature-dependent molecular diversity and dynamic formation of smoke-derived WSOC remain poorly understood in water. Herein, we explored the molecular-level formation mechanism of pyrolytic smoke-derived WSOC in water to explain the evolution, heterogeneous correlations, and sequential responses of molecules and functional groups to increasing pyrolysis temperature. Two-dimensional correlation spectroscopy was used to innovatively establish the characteristic correlations between spectroscopy and Fourier transform-ion cyclotron resonance mass spectrometry. Temperature-dependent formation of WSOC exhibited diversity in absorbance/fluorescent components, unique/common molecules, and their chemical parameters, showing the simultaneous formation and degradation reactions. The common WSOC molecules with lower and higher degrees of oxidation showed significant positive and negative correlations with the fluorescent components, respectively. The primary sequential response of WSOC molecules to increasing pyrolysis temperature (lignin-like molecules → unsaturated hydrocarbons, condensed aromatic molecules → lipid-like/aliphatic-/peptide-like molecules) corresponded to the temperature response of functional groups (carboxylic/alcoholic → polysaccharides → aromatics/amides/phenolic/aliphatic groups), demonstrating well synergistic relationships between them. These novel findings will contribute to the comprehensive understanding and assessments of potential environmental behavior or risks of WSOC in aquatic ecosystems.

Emerging investigator series: impacts of land use on dissolved organic matter quality in agricultural watersheds: a molecular perspective.

In aquatic systems, dissolved organic matter (DOM) has important ecological and biogeochemical functions, where the molecular composition of DOM has larger-scale implications for climate change and global carbon cycles. However, there is limited information about the relationships between landscape characteristics and human disturbance that influence the molecular composition of DOM changes in watersheds. In this study, we collected water samples from 22 sites across a gradient of topographically characterized agricultural land coverage and community infrastructure development in the Kawartha region in Ontario, Canada. We employed a combination of Fourier Transform Ion-Cyclotron Resonance Mass Spectrometry (FT-ICR-MS) and absorbance spectroscopy to investigate changes in the molecular composition of DOM with increasing agricultural and community development disturbance on the optical and molecular characteristics of DOM. We found that dissolved organic carbon (DOC) concentrations in disturbed (>75%) watersheds ranged from 3.67-32.8 mg L-1 and were significantly higher than in watersheds with more abundant forest coverage (3.78-9.13 mg L-1). In addition, watersheds with higher phosphorus concentrations had more negative nominal oxygenation state of carbon (NOSC) values, suggesting biologically processed DOM correlating with increased phosphorus levels in aquatic systems. To relate the molecular properties of DOM to landscape metrics, we used Spearman's correlation analysis to reveal that agriculturally impacted and community developments enhanced the molecular signature of unsaturated hydrocarbon. In addition, we identified 65 dissolved organic phosphorus (DOP) molecules that significantly increased in abundance with disturbance, likely due to microbial mineralization of existing DOM with the addition of phosphorus to form larger, biologically inaccessible molecules. The overall recalcitrance of the identified molecules can serve as molecular signatures when evaluating the level of disturbance of a watershed.

Characterization and surrogate formulation of heavy fuel oil

Residual fuel oils have recently garnered attention as promising feedstocks for gasification processes. However, their complex compositions pose challenges for accurate characterization and simulation. This study addresses this issue by formulating a surrogate molecule for heavy fuel oil (HFO) with minimal components, aiming to replicate the average molecular formula and functional groups distribution found in the original fuel. To overcome challenges associated with Fourier transform-ion cyclotron resonance mass spectrometry (FT-ICR MS) detection limits, a distillation process was employed, yielding two fractions: residual distillate (enriched in heavy complex components) and volatile distillate (enriched in light components). Both fractions undergo comprehensive characterization through nuclear magnetic resonance (NMR) spectroscopy, FT-ICR MS for the residual distillate and gas chromatography-mass spectrometry (GC–MS) for the volatile distillate.The study highlights the importance of estimating average molecular weight (AMW) using both fractions to avoid overestimation when relying solely on FT-ICR MS. The developed six-component surrogate, based on elemental analysis, average molecular parameters (AMPs), and AMW of each fraction, demonstrates accuracy in replicating some properties of the original HFO. The surrogate proves effective compared to quantitative structure-property relationship (QSPR) calculations and holds potential for simulating HFO in gasification processes. The molecular design of the surrogate incorporates 1H and 13C NMR data, ensuring similarity in AMPs, AMW, and molecular formula to the target HFO. This comprehensive approach enhances the understanding and application of surrogate molecules for simulating residual oils in gasification processes.

Low-temperature oxidation characteristics and reaction pathways of crude oil within tight shale during air injection

The investigation of low-temperature oxidation (LTO) of crude oil within tight shale holds significant importance due to its implications for subsequent oxidation reactions and enhanced oil recovery in the process of air injection. In this study, the tight shale sample underwent oxidation at various LTO temperatures, followed by an analysis of the resulting gas composition. Furthermore, the oxidized oil was separated from the tight shale and subjected to characterization using electron paramagnetic resonance, nuclear magnetic resonance, and negative ion electrospray Fourier transform-ion cyclotron resonance mass spectrometry techniques. The primary focus was on examining the distinct LTO reaction pathways observable across different temperature ranges. The findings demonstrated a correlation between LTO temperature and the concentration of free radicals, which predominantly resided on aromatic hydrocarbons, alkanes, and oxygen atoms. Additionally, the proton count of polycyclic aromatic hydrocarbons exhibited a continuous increase from 83 to 350 °C, suggesting intensified aromatization and condensation reactions involving aliphatic and aromatic compounds. With rising LTO temperature, the molecular structure of O2 compounds underwent significant transformations, characterized by increased condensation degree and a decrease in low carbon number molecular structures, while higher equivalent double bonds and carbon number molecular structures became more prevalent. The influence of cycle path 1 diminished at temperatures ranging from 83 to 150 °C and 250 to 350 °C, whereas the significance of cycle paths 2 and 3 increased, resulting in an overall escalation of the oxidation rate with temperature elevation. It was observed that the shale oil LTO process exhibited a negative temperature coefficient within the temperature range of 150-250 °C, emphasizing the criticality of overcoming the energy barrier in this region to achieve stable combustion. This comprehensive investigation provides valuable insights into the mechanisms underlying LTO in crude oil confined within tight shale.

Comparison of secondary organic aerosol (SOA)-associated molecular features at urban sites in China and Korea in winter and summer (2019)

Secondary organic aerosol (SOA) formation is important in the field of atmospheric science due to its impacts on public health and climate change. However, the molecular compositions or formation mechanisms of SOA have not been fully characterized in ambient atmospheres. In this study, ambient fine particle (PM2.5) samples were collected in two urban cities in Beijing, China and Gwangju, Korea during the winter and summer seasons, and the SOA-associated molecular features were characterized by using Fourier transform-ion cyclotron resonance mass spectrometry (FT-ICR MS). We found that the secondary organic carbon (SOC) during wintertime in Gwangju exhibit a more aliphatic nature, and those in Beijing have more aromatic structures like humic compounds. The contribution of CHOS to the total identified WSOC associated SOC was the most abundant at Beijing site (∼40%) compared to the Gwangju site (∼30%), and the proportion of CHON species in the WSOC associated SOC accounted for ∼30% in Beijing and 20–30% in Gwangju. Majority of SOC-derived CHOS species (∼99%) likely contained sulfate functional groups at both sites regardless of the sampling season. Organonitrates predominated in CHON species in summer compared to winter, but some fractions appeared to contain other potential nitrogen functional groups such as amide and nitrile (∼20%), suggesting the existence of other formation pathways of organic nitrogen, especially during winter. This work gave an observation insight into the molecular features of SOC at two different sites, and will deepen the understanding on the region-specific SOA formation mechanisms.

Characterization of polycyclic aromatic hydrocarbons (PAHs) in atmospheric particles (PM2.5) in the North China Plain using positive-ion atmospheric pressure photoionization (APPI) coupled with fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS)

Many aromatic compounds, including 16 types of U.S. EPA priority PAHs, exist in substituted or unsubstituted forms and have high toxicities. Aromatic compounds in aerosols collected in the North China Plain (NCP) were characterized by GC-MS and atmospheric pressure photoionization (APPI) Fourier transform-ion cyclotron resonance mass spectrometry (FT-ICR MS). The concentrations of the 16 types of PAHs in the collected samples ranged from 277.0 to 876.8 ng/m3, which were higher than those collected in other regions and countries. Previous studies reported that only 27 wt% of aromatic compounds could be eluted using the GC system and demonstrated that APPI FT-ICR MS is a useful method for identifying large-molecular aromatics. In this study, two samples collected from the NCP were successfully characterized by APPI FT-ICR MS. In the APPI + mode, 231 and 228 CH formulas (containing only C and H and having a double bond equivalent (DBE) of 4 or greater) were identified, accounting for 10% of the numerical fraction and more than 40% of the intensity fraction. The FT-ICR data revealed that condensed PAHs with 6–8 aromatic rings had relatively high abundances and that a series of PAH homologs (alkylated PAHs) with identical DBE values ranging from 4 to 30 were present. Furthermore, heteroatom groups were also identified as significant components of the aromatic fractions. The O1, O2, N1, NO2, and S1 class species with DBE values of 3–34 and carbon numbers of 11–42 were identified by APPI FT-ICR MS. The heteroatom PAHs had DBE values and carbon number distributions similar to regular PAHs. The aromatic fractions determined in this work provide useful data for future quantitative analyses of aromatic compounds with high toxicities and our findings will help improve the current understanding of PAH compositions in the northern China. However, further research is needed to determine the abundances and toxicities of various apparent or substituted PAHs outside of the 16 types identified by the U.S. EPA as priority PAH compounds, which have been extensively researched.

A Research Journey: Over a Decade of Denaturing and Native-MS Analyses of Hydrophobic and Membrane Proteins in Amgen Therapeutic Discovery.

Membrane proteins and associated complexes currently comprise the majority of therapeutic targets and remain among the most challenging classes of proteins for analytical characterization. Through long-term strategic collaborations forged between industrial and academic research groups, there has been tremendous progress in advancing membrane protein mass spectrometry (MS) analytical methods and their concomitant application to Amgen therapeutic project progression. Herein, I will describe a detailed and personal account of how electrospray ionization (ESI) native mass spectrometry (nMS), ion mobility-MS (IM-MS), reversed phase liquid chromatographic mass spectrometry (RPLC-MS), high-throughput solid phase extraction mass spectrometry, and matrix-assisted laser desorption ionization mass spectrometry methods were developed, optimized, and validated within Amgen Research, and importantly, how these analytical methods were applied for membrane and hydrophobic protein analyses and ultimately therapeutic project support and progression. Additionally, I will discuss all the highly important and productive collaborative efforts, both internal Amgen and external academic, which were key in generating the samples, methods, and associated data described herein. I will also describe some early and previously unpublished nano-ESI (nESI) native-MS data from Amgen Research and the highly productive University of California Los Angeles (UCLA) collaboration. I will also present previously unpublished examples of real-life Amgen biotherapeutic membrane protein projects that were supported by all the MS (and IM) analytical techniques described herein. I will start by describing the initial nESI nMS experiments performed at Amgen in 2011 on empty nanodisc molecules, using a quadrupole time-of-flight MS, and how these experiments progressed on to the 15 Tesla Fourier transform ion cyclotron resonance MS at UCLA. Then described are monomeric and multimeric membrane protein data acquired in both nESI nMS and tandem-MS modes, using multiple methods of ion activation, resulting in dramatic spectral simplification. Also described is how we investigated the far less established and less published subject, that is denaturing RPLC-MS analysis of membrane proteins, and how we developed a highly robust and reproducible RPLC-MS method capable of effective separation of membrane proteins differing in only the presence or absence of an N-terminal post translational modification. Also described is the evolution of the aforementioned RPLC-MS method into a high-throughput solid phase extraction MS method. Finally, I will give my opinion on key developments and how the area of nMS of membrane proteins needs to evolve to a state where it can be applied within the biopharmaceutical research environment for routine therapeutic project support.

Molecular insight into DOM fate using EEM-PARAFAC and FT-ICR MS and concomitant heavy metal behaviors in biologically treated landfill leachate during coagulation: Al speciation dependence

Various combined processes with pre-coagulation have been developed for biologically treated landfill leachate, but the microscopic-level processes occurring during coagulation remain largely unknown. Herein, dissolved organic matter (DOM) fate using fluorescence excitation emission matrix spectroscopy combined with parallel factor analysis and electrospray ionization coupled Fourier transform-ion cyclotron resonance mass spectrometry and concomitant heavy metal (HM) behaviors were explored at the molecular level. In addition, AlCl3 and two polyaluminum chloride (PACl) species (dominated by [AlO4Al12(OH)24(H2O)12]7+ and [(AlO4)2Al28(OH)56(H2O)26]18+, respectively) were used. The results show that all coagulants are efficient at removing DOM. PACl was found to be advantageous over AlCl3 in overcoming pH fluctuation, which is ascribed to the different dominant mechanisms, namely, entrapment and sweep flocculation for AlCl3 and charge neutralization for PACl. Consequently, PACl was more effective at removing humic substances, usually high-molecular-weight, oxygen-rich and unsaturated, than protein substances. For HM removal, PACl was likewise better and more stable, where As, Cu, Ni, Co and Hg were removed predominantly via adsorption, and Cr, Zn, Pb, Cd and Mn were removed via coprecipitation. Correlation analysis showed that humic substances tended to complex with HMs and be removed synergistically due to the ubiquitous occurrences of aromatic structures and oxygen-containing functional groups.

Mechanistic Insights into the Biodegradation of Carbon Dots by Fungal Laccase.

While carbon dots (CDs) have the potential to support the agricultural revolution, it remains obscure about their environmental fate and bioavailability by plants. Fungal laccase-mediated biotransformation of carbon nanomaterials has received little attention despite its known capacity to eliminate recalcitrant contaminants. Herein, we presented the initial investigation into the transformation of CDs by fungal laccase. The degradation rates of CDs were determined to be first-order in both substrate and enzyme. Computational docking studies showed that CDs preferentially bonded to the pocket of laccase on the basal plane rather than the edge through hydrogen bonds and hydrophobic interactions. Electrospray ionization-Fourier transform-ion cyclotron resonance mass spectrometry (ESI-FT-ICR MS) and other characterizations revealed that the phenolic/amino lignins and tannins portions in CDs are susceptible to laccase transformation, resulting in graphitic structure damage and smaller-sized fragments. By using the 13C stable isotope labeling technique, we quantified the uptake and translocation of 13C-CDs by mung bean plants. 13C-CDs (10 mg L-1) accumulated in the root, stem, and leaf were estimated to be 291, 239, and 152 μg g-1 at day 5. We also evidenced that laccase treatment alters the particle size and surface chemistry of CDs, which could facilitate the uptake of CDs by plants and reduce their nanotoxicity to plants.

New Insights into the Role of Natural Organic Matter in Fe-Cr Coprecipitation: Importance of Molecular Selectivity.

Coprecipitation of Fe/Cr hydroxides with natural organic matter (NOM) is an important pathway for Cr immobilization. However, the role of NOM in coprecipitation is still controversial due to its molecular heterogeneity and diversity. This study focused on the molecular selectivity of NOM toward Fe/Cr coprecipitates to uncover the fate of Cr via Fourier transform-ion cyclotron resonance-mass spectrometry (FT-ICR-MS). The results showed that the significant effects of Suwannee River NOM (SRNOM) on Cr immobilization and stability of the Fe/Cr coprecipitates did not merely depend on the adsorption of SRNOM on Fe/Cr hydroxides. FT-ICR-MS spectra suggested that two pathways of molecular selectivity of SRNOM in the coprecipitation affected Cr immobilization. Polycyclic aromatics and polyphenolic compounds in SRNOM preferentially adsorbed on the Fe/Cr hydroxide nanoparticles, which provided extra binding sites and promoted the aggregation. Notably, some specific compounds (i.e., polyphenolic compounds and highly unsaturated phenolic compounds), less unsaturated and more oxygenated than those adsorbed on Fe/Cr hydroxide nanoparticles, were preferentially incorporated into the insoluble Cr-organic complexes in the coprecipitates. Kendrick mass defect analysis revealed that the insoluble Cr-organic complexes contained fewer carbonylated homologous compounds. More importantly, the spatial distribution of insoluble Cr-organic complexes was strongly related to Cr immobilization and stability of the Fe/Cr-NOM coprecipitates. The molecular information of the Fe/Cr-NOM coprecipitates would be beneficial for a better understanding of the transport and fate of Cr and exploration of the related remediation strategy.