Journal of the American Society for Mass Spectrometry最新文献

{"title":"Desorption Electrospray Ionization-Mass Spectrometry Imaging Provides Spatiochemical Information on Potential Biocontrol Agents against <i>Phytophthora capsici</i> Infection in Tomato Plants.","authors":"Jamille Y Robinson, Hawkins S Shepard, Daniel Ambachew, Peter J Eyegheleme, Jody C May, Margaret T Mmbaga, John A McLean","doi":"10.1021/jasms.6c00067","DOIUrl":"https://doi.org/10.1021/jasms.6c00067","url":null,"abstract":"<p><p>Biological control agents can offer an eco-friendly and more sustainable alternative to conventional chemical pesticides, providing protection against destructive pathogens, such as <i>Phytophthora capsici</i>, while reducing potential environmental harm associated with synthetic pesticide use in agricultural systems. This work evaluates the biocontrol effectiveness of <i>Bacillus vallismortis</i>, <i>Bacillus amyloliquefaciens</i>, <i>Bacillus thuringiensis</i>, and <i>Bacillus subtilis</i>, against the widespread plant pathogen <i>Phytophthora capsici</i>. Our studies showed that <i>Bacillus thuringiensis</i> and <i>Bacillus subtilis</i> promote plant growth and provide protection against <i>Phytophthora capsici</i> in both in vitro and in vivo greenhouse studies, while <i>Bacillus vallismortis</i> and <i>Bacillus amyloliquefaciens</i> were effective in vitro but not in vivo. Specifically, <i>Bacillus thuringiensis</i> was observed to both hinder the growth of <i>Phytophthora capsici</i> and enhance plant resilience to this pathogen, with <i>B. thuringiensis</i><i>-</i>treated, pathogen-exposed plants displaying a 94.4% increase in root length and a 74.0% increase in shoot height compared to plants with only oomycete exposure. To probe the molecular interactions between the biocontrol agent and pathogen, a dual culture of <i>Bacillus thuringiensis</i> and <i>Phytophthora capsici</i> was analyzed in situ using a desorption electrospray ionization-mass spectrometry imaging (DESI-MSI) workflow. This approach interrogated the spatially oriented biochemical interactions that may serve as the molecular foundation for the effectiveness of these biological control agents in crop protection, identifying seven unique phenotypic regions within the dual culture. Herein, we demonstrate the benefits of biological control agent application in tomato cultivation and showcase the strengths of desorption electrospray ionization-mass spectrometry imaging when applied to the spatially resolved molecular characterization of agriculturally relevant microorganisms.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":""},"PeriodicalIF":2.7,"publicationDate":"2026-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147832052","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Evaluating the Sulfo-Phospho-Vanillin Assay for Total Lipid Sample Normalization as Compared to Gravimetric and Protein Sample Normalization Methods for Untargeted Lipidomic LC–MS/MS","authors":"Laura S. Bailey,&nbsp; and ,&nbsp;Kari B. Basso*,&nbsp;","doi":"10.1021/jasms.5c00429","DOIUrl":"10.1021/jasms.5c00429","url":null,"abstract":"<p >Sample normalization is essential for lipid quantitation. While normalization methods involving pre- or postgravimetric measurements, counting, or protein prequantitation exist, a single, common sample normalization method is not widely accepted. Previously, we proposed and evaluated the sulfo-phospho-vanillin assay (SPVA), a total lipid quantitation reaction, for prequantifying total lipids for LC–MS/MS sample normalization purposes. This current investigation furthers our evaluation of the SPVA as a sample normalization method in untargeted lipidomic LC–MS/MS by comparing SPVA total lipid prequantitation to protein prequantitation and gravimetric measurements. This study was applied to a wide selection of matrices, including <i>Escherichia coli</i>, plasma, brain, heart, and kidney. Resulting relative lipid concentrations showed smaller bioreplica variation when a prequantitation method was applied, either measuring total lipid or total protein; however, several relative lipid abundances showed inverse concentration relationships when normalizing with total protein compared to total lipid (from SPVA measurements) or gravimetric sample normalization. Further investigation using lipid extracts linearly spiked with pure, non-native lipid showed that gravimetric and protein normalization nonlinearly overproduced significant lipids, while linear increases in significant lipid features were observed from SPVA normalization. Lipid extracts linearly spiked with pure, non-native protein yielded fewer significant lipid features using SPVA normalization, and this was unchanged as protein was added; however, both gravimetric and protein normalization continued to yield large numbers of significant lipid features. Together, these results suggest neither gravimetric nor protein sample normalization appropriately normalizes quantitative lipidomic experiments and greatly overgenerates statistically significant lipid features for biomarker investigation. SPVA normalization more accurately adjusts for lipid changes in bioreplicate samples, leading to fewer but more biologically relevant statistically significant lipid features.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":"37 5","pages":"1149–1159"},"PeriodicalIF":2.7,"publicationDate":"2026-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147472291","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Investigating the Efficiency of Ultraviolet Photodissociation in Peptides Modified with N-Terminal UV-Absorbing Chromophores","authors":"Nikita Levin,&nbsp;, ,&nbsp;Yana Demyanenko,&nbsp;, ,&nbsp;Eduardo Kitano,&nbsp;, and ,&nbsp;Shabaz Mohammed*,&nbsp;","doi":"10.1021/jasms.5c00389","DOIUrl":"10.1021/jasms.5c00389","url":null,"abstract":"<p >Ultraviolet photodissociation (UVPD) has emerged as a powerful alternative to conventional collision-induced dissociation (CID) for peptide and protein sequencing in mass spectrometry-based proteomics. However, the UVPD efficiency depends on the presence of UV-absorbing chromophores within analytes. Here, we systematically investigate how <i>N</i>-terminal modification of peptides with aromatic chromophores influences the 193 nm UVPD efficiency and fragmentation behavior. Using NHS ester chemistry, human cell lysate digests were derivatized with a range of UV-absorbing aromatic labels and analyzed on an Omnitrap–Orbitrap platform equipped with a 193 nm ArF excimer laser. Compared with unmodified controls, derivatized peptides displayed a modestly increased fragmentation efficiency and a greater diversity of sequence-informative fragments, particularly <i>b</i>-type ions. Peptides lacking aromatic amino acids benefited most from N-terminal aromatic labeling, exhibiting enhanced fragmentation and sequence coverage. These findings demonstrate that incorporation of aromatic chromophores at the N-terminus can enhance UVPD performance and improve spectral interpretability, thereby expanding the applicability of UVPD in bottom-up proteomics workflows.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":"37 5","pages":"1124–1131"},"PeriodicalIF":2.7,"publicationDate":"2026-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/jasms.5c00389","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147508591","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Experimental and Computational Investigation of Sulfur Chemistry in the DART-MS Gas Stream: Implications for the Interpretation of the Mass Spectra of Organic Disulfides","authors":"Benedetta Garosi,&nbsp;, ,&nbsp;Parandaman Arathala,&nbsp;, and ,&nbsp;Rabi A. Musah*,&nbsp;","doi":"10.1021/jasms.6c00046","DOIUrl":"10.1021/jasms.6c00046","url":null,"abstract":"<p >Direct analysis in real time–high-resolution mass spectrometry (DART-HRMS) has proven useful for the detection and faithful representation of labile organosulfur compounds (OSCs). Nevertheless, it has been found that when exposed to the metastable helium (He*) of the DART gas stream under soft ionization conditions, some classes of OSCs such as disulfides, undergo several reactions to produce new organosulfur species, which complicate interpretation of their spectra. In this work, these new entities were characterized and the mechanisms of their formation explored. DART-HRMS analysis of diphenyl disulfide (PhSSPh) exhibited peaks consistent with [PhSSPh]^+•, [PhSSPh + H]⁺, [PhSPh]^+•, and [PhSOSPh + H]⁺ and various fragments and adducts including [PhSS]⁺ and [Ph₃S₃]⁺. <i>Ab initio</i>/DFT calculations coupled with variational transition state theory revealed that several of these peaks are artifacts of reactions occurring with He* and ^•OH. Branching ratio analysis showed the dominant decomposition pathway of [PhSSPh]^+• to be [PhSSPh]^+• → PhS^• + PhS:, rather than [PhSSPh]^+• → PhS⁺=S + Ph^•. The PhS^•, Ph^•, and ^•OH formed serve as key intermediates in subsequent reactions that lead to the <i>m</i>/<i>z</i> values observed. From the results, a systematic approach for the interpretation of the DART mass spectra of organic disulfides was developed and successfully applied to various disulfide compound classes (e.g., aryl, alkyl, benzyl, and phenyl). Organic solvents were also observed to influence the ability to detect the compounds. Benzene; dichloromethane; hexane; and with some exceptions, ethyl acetate and tetrahydrofuran were found to be suitable, while disulfide signals in dimethylformamide were totally absent.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":"37 5","pages":"1268–1280"},"PeriodicalIF":2.7,"publicationDate":"2026-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/jasms.6c00046","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147687661","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Native Top-Down Mass Spectrometry Combined with High-Resolution Charge Variant Analysis of Trastuzumab Originator and Biosimilars","authors":"Corentin Beaumal,&nbsp;, ,&nbsp;Kristina Srzentić,&nbsp;, ,&nbsp;Sara Carillo,&nbsp;, and ,&nbsp;Jonathan Bones*,&nbsp;","doi":"10.1021/jasms.5c00438","DOIUrl":"10.1021/jasms.5c00438","url":null,"abstract":"<p >Comprehensive characterization of monoclonal antibody (mAb) charge heterogeneity is essential to ensure product quality, maintain batch consistency and support biosimilar development. Charge variant analysis (CVA) is widely used to separate acidic and basic proteoforms from the main species. However, cation-exchange chromatography coupled to mass spectrometry provides limited information and cannot localize the post-translational modifications (PTMs) responsible for mAb heterogeneity. Here, the coupling of pH-gradient CVA with native top-down mass spectrometry (TD-MS) for proteoform-specific analysis of trastuzumab is presented. Individual charge variants were chromatographically separated under native conditions and directly fragmented on the chromatographic time scale using higher-energy collision dissociation (HCD), electron-transfer dissociation (ETD) and ultraviolet photodissociation (UVPD). The addition of proton-transfer charge reduction (PTCR) helped reduce spectral congestion and enhanced the detection of high-mass fragment ions, resulting in improved sequence coverage. This workflow enabled the complete sequencing of the complementarity-determining region (CDR) 3 and the direct identification and insights into the location of key PTMs at the intact-protein level, including deamidation, succinimide and N-terminal pyroGlu for individual proteoforms. Comparison of five trastuzumab samples (originator and biosimilars) demonstrated high reproducibility in fragmentation patterns, sequence coverage and variant assignment, highlighting the robustness of the method. Although limitations remain due to the challenges of fragmenting intact mAbs under native conditions, this work establishes a proof of concept for CVA native TD-MS characterization of mAbs to complement bottom-up and middle-down analyses, and has potential for broad applicability for antibody-based biopharmaceuticals.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":"37 5","pages":"1173–1186"},"PeriodicalIF":2.7,"publicationDate":"2026-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/jasms.5c00438","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147715568","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Unsupervised Semantic Segmentation Models for Region of Interest Identification","authors":"David J. Degnan,&nbsp;, ,&nbsp;Bailey G. Knight,&nbsp;, ,&nbsp;Logan A. Lewis,&nbsp;, ,&nbsp;Ian A. VanGordon,&nbsp;, ,&nbsp;Teminijesu T. Adewunmi,&nbsp;, ,&nbsp;Lauren M. Jones,&nbsp;, ,&nbsp;Shruthilayaa Sivakumar,&nbsp;, ,&nbsp;Rashmi Kumar,&nbsp;, ,&nbsp;Dušan Veličković,&nbsp;, ,&nbsp;Kevin J. Zemaitis,&nbsp;, ,&nbsp;Ljiljana Paša-Tolic,&nbsp;, and ,&nbsp;Lisa M. Bramer*,&nbsp;","doi":"10.1021/jasms.5c00216","DOIUrl":"10.1021/jasms.5c00216","url":null,"abstract":"<p >Spatial omics technologies, such as mass spectrometry imaging (MSI), can capture biomolecular distributions and their spatial locations directly from a tissue, but these distributions are not easily associated with tissue morphology without additional microscopy performed on the sample. To identify tissue regions of interest (ROIs), a reference image (such as a stained tissue, microscopy images, etc.) may be used and paired with mass spectrometry data. Due to the intense time requirements of manually labeling ROIs, segmentation models have been demonstrated as time-saving alternatives, as they automatically annotate ROIs by grouping like pixel intensities together. Supervised approaches trained on specific image types are preferred, but when those cases are not available, generalizable unsupervised segmentation models may then be used. Here, eight unsupervised semantic segmentation algorithms in R and Python, representing both statistical and machine learning algorithms, were compared for their ability to match manual annotations of 30 tiled PAS-stained kidney images and 25 tiled plant root images. Noise reduction techniques such as dimension reduction were tested to see whether they improved segmentations, and all models were applied to the full stitched images. Performance metrics were calculated to provide recommendations on the highest performing models to demonstrate their potential for automated annotation of tissue ROIs. At small cluster sizes, k-means and pytorch-tip tended to have the best performance in terms of balanced accuracy and time, though all algorithms had decreased performance at higher cluster numbers. Lastly, the segmentation model choice was demonstrated to have an impact on downstream statistics, highlighting the importance of testing and selecting the best segmentation model on a case-by-case basis, as no one model had the best performance in every comparison.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":"37 5","pages":"1068–1076"},"PeriodicalIF":2.7,"publicationDate":"2026-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147759211","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Sensitive RP-LC Method Enabling PTM-Specific Quality Control and MS-Compatible Characterization of Fc-Containing GLP-1 Therapeutics.","authors":"Jin Xu, Anqi Zhou, Mengjiao Xu, Qiaoshuang Chen, Chenjing Xu, Jinhao Li, Na Li, Zhixin Li, Guodong Wu, Weitao Zhang, Shuhai Xu, Qiaoling Ni, Tao Liu, Chunguang Zheng, Wei Yu, Xi Chen, Lankun Song, Min Xia, Qingcheng Guo, Lusha Ji, Weizhu Qian, Sheng Hou, Jun Li, Ziqiao Sun, Xuekun Wang, Fubo Han, Hao Wang, Jing Li, Dapeng Zhang, Huaizu Guo","doi":"10.1021/jasms.6c00052","DOIUrl":"https://doi.org/10.1021/jasms.6c00052","url":null,"abstract":"<p><p>Fc-containing GLP-1 therapeutics exhibit complex post-translational modification (PTM) heterogeneity, necessitating advanced analytical methods for quality control (QC) and process analytical technology (PAT). We developed a reverse-phase liquid chromatography (RP-LC) method for the PTM-specific profiling of these biologics. Using dulaglutide (IgG4-Fc) as a model, critical parameters─including mobile-phase additives, acid concentration, and shallow gradients─were optimized to resolve PTM variants (e.g., hydroxylation, N-terminal truncation, disulfide reduction, glycosylation) within 40 min. Mass spectrometry (MS) compatibility was enabled by adopting difluoroacetic acid (DFA) as an alternative ion-pairing reagent to support intact-mass characterization of variants. This enabled the identification of additional PTMs not readily resolved under the initial RP-LC conditions, including site-specific HyK-Gal-Glc O-glycosylation and process-dependent truncations. The method also allowed for the direct quantification of critical impurities and the detection of process-induced variants across biosimilar clones. The method was further demonstrated on the IgG2-subtype GLP-1-Fc-fusion (supaglutide), showing applicability across the two Fc subtypes examined without additional optimization. This robust, MS-compatible RP-LC platform provides a rapid, accurate, and comprehensive (RAC) means of conducting PTM-specific QC for Fc-GLP-1 therapeutics, supporting PAT implementation and accelerating biosimilar and next-generation drug development.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":" ","pages":""},"PeriodicalIF":2.7,"publicationDate":"2026-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147832064","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Improved Methods for Recording Accurate Collision Cross Sections Using Cyclic Ion Mobility-Mass Spectrometry","authors":"Devin M. Makey,&nbsp;, ,&nbsp;David Langridge,&nbsp;, ,&nbsp;Keith Richardson,&nbsp;, ,&nbsp;Ryan Schroeder,&nbsp;, ,&nbsp;Jakub Ujma,&nbsp;, ,&nbsp;Kevin Giles,&nbsp;, and ,&nbsp;Brandon T. Ruotolo*,&nbsp;","doi":"10.1021/jasms.6c00022","DOIUrl":"10.1021/jasms.6c00022","url":null,"abstract":"<p >Traveling wave ion mobility (TWIM) is a gas-phase separation technique widely used in structural biology to rapidly probe protein conformational states and in complex mixture analysis to catalyze an array of chemical measurements. Analyte size information can be extracted from TWIM data as collision cross section (CCS) values, which are physicochemical descriptors of the analytes. However, calibration using ions of known CCS is currently required. Recently, an improved TWIM calibration approach was introduced that incorporates blend and radial expressions derived from velocity relaxation and axial confinement effects. This approach offers several benefits over conventional power-law calibrations, including accurate calibration over a wide range of separation conditions and analyte classes using a universal calibrant set. While the blend and radial calibrations were extensively validated using a linear TWIM separator, next-generation cyclic ion mobility (cIM) offers additional benefits, such as scalable resolution using multipass separations and the ability to subject ions to multiple stages of activation and separation (i.e., IMSⁿ). The original work provided a brief application of the blend and radial expressions to cIM calibrations, but these cIM CCS calibrations were applied only to a basic single-pass mode of operation under limited separation conditions. Here, we describe a comprehensive method for accurate cIM calibrations using blend and radial expressions. Correcting ion arrival times for the time spent outside the separator and using conditions that minimize velocity relaxation effects reduced CCS measurement error. We also provide an optimized workflow for calibration using the blend and radial expressions in multipass and IMSⁿ modes of operation.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":"37 5","pages":"1237–1244"},"PeriodicalIF":2.7,"publicationDate":"2026-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147589363","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Avoiding False Identification of 7-Hydroxymitragynine in Kratom Products Using a Multicriteria LC–MS Confirmation","authors":"Daniel Sheehan,&nbsp;, ,&nbsp;Yanfang Li,&nbsp;, ,&nbsp;Emily Meckler,&nbsp;, ,&nbsp;Roy Upton,&nbsp;, and ,&nbsp;Mengliang Zhang*,&nbsp;","doi":"10.1021/jasms.6c00088","DOIUrl":"10.1021/jasms.6c00088","url":null,"abstract":"<p >Kratom (<i>Mitragyna specios</i>a) products have become widely available in diverse commercial formulations, raising increasing analytical and regulatory interest due to the presence of potent alkaloids such as mitragynine (MG), 7-hydroxymitragynine (7-OH), and mitragynine pseudoindoxyl (MGP). The accurate identification of these compounds is analytically challenging because kratom matrices contain numerous structurally related alkaloids and isomeric species that produce similar mass spectral features. In this study, an ultrahigh-performance liquid chromatography–high-resolution mass spectrometry (UHPLC–HRMS) workflow was developed to characterize kratom alkaloids in authentic leaf materials and 38 commercial kratom products representing multiple formulations. Chromatographic separation combined with accurate mass measurement and diagnostic MS/MS fragmentation enabled reliable differentiation of MG, 7-OH, MGP, and related isomers. Multiple overlapping chromatographic peaks were observed at <i>m</i>/<i>z</i> 415.2227, demonstrating that reliance solely on accurate mass or commonly monitored transitions such as <i>m</i>/<i>z</i> 415 → 190 can lead to false-positive identification of 7-OH in complex botanical matrices. Application of the multicriteria confirmation strategy revealed substantial variability in alkaloid composition across commercial products, including the presence of 7-OH and MGP in several tablet and capsule formulations. Untargeted metabolomic profiling further distinguished kratom plant extracts, including the authentic leaf materials, from processed products enriched in specific alkaloids. These results highlight the importance of integrating chromatographic resolution with orthogonal MS/MS criteria for reliable identification of kratom alkaloids and provide an analytical framework for the characterization of complex botanical products.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":"37 5","pages":"1303–1312"},"PeriodicalIF":2.7,"publicationDate":"2026-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/jasms.6c00088","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147697529","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Quantitative Host Cell Protein Analysis of Antibody-Based Protein Therapeutics Using the Orbitrap Astral Mass Spectrometer","authors":"Josh Smith,&nbsp;, ,&nbsp;Aaron Richardson,&nbsp;, ,&nbsp;Corentin Beaumal,&nbsp;, ,&nbsp;Marina Ainciburu,&nbsp;, ,&nbsp;Sara Carillo,&nbsp;, ,&nbsp;Anna Pashkova,&nbsp;, ,&nbsp;Tabiwang N. Arrey,&nbsp;, ,&nbsp;Nicolaie E. Damoc,&nbsp;, ,&nbsp;Colin Clarke,&nbsp;, and ,&nbsp;Jonathan Bones*,&nbsp;","doi":"10.1021/jasms.5c00272","DOIUrl":"10.1021/jasms.5c00272","url":null,"abstract":"<p >Host cell proteins (HCPs) are endogenous proteins generated in cellular production systems alongside the biotherapeutic of interest. Removal of HCPs is crucial as they can be detrimental to product efficacy and patient safety. Due to its ability to determine individual HCP concentrations, liquid chromatography tandem mass spectrometry is increasingly utilized as an orthogonal method to ELISA for HCP monitoring. For protein biotherapeutics like monoclonal antibodies, their dynamic range makes detection of low-level HCPs difficult. The Orbitrap Astral MS has the potential to overcome such challenges, offering improvements in protein identifications in complex sample matrices while simultaneously reducing analysis times. Here, we utilize the Orbitrap Astral MS to perform HCP analysis on 36 protein biotherapeutics. Our workflow used a short 60 samples-per-day separation method and was initially benchmarked against four previously published studies, demonstrating comparable levels of HCP identifications. 236 HCPs were detected across the cohort and 55% of those found to be quantifiable in at least one product using label free quantitation. Functional analysis revealed that most detected HCPs had functions related to catalysis or binding, predominately catalytic activity (46%, 97 gene IDs) or protein binding (44%, 91 gene IDs). Nearly 80% of quantifiable HCPs were detected at concentrations below 10 ng/mg, with 8% detected below concentrations of 1 ng/mg. These included HCPs considered as “high-risk” by the Biophorum Development Group. This study shows how new generation mass spectrometry instruments can enable detection of low-level HCPs while allowing for a rapid and more informed understanding of a product’s HCP content.</p>","PeriodicalId":672,"journal":{"name":"Journal of the American Society for Mass Spectrometry","volume":"37 5","pages":"1077–1094"},"PeriodicalIF":2.7,"publicationDate":"2026-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/jasms.5c00272","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147715572","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}