<p><strong>🎤Dr. Christopher Reid (Professor of Chemistry and Biochemistry at Bryant University in Rhode Island)</strong></p><p><strong>In this episode of Concentrating on Chromatography, host Dave Oliva sits down with Chris Reid to explore how chemical biology and LC-MS are being used to tackle one of the most urgent global challenges: antimicrobial resistance.</strong></p><p>As antibiotic resistance continues to rise—and new drug development struggles to keep pace—researchers are looking beyond traditional approaches. Instead of targeting how bacteria build their cell walls, Chris’s work focuses on how they break them down, opening the door to entirely new antimicrobial strategies.</p><h4><strong>What You’ll Learn in This Episode:</strong></h4><ul><li>Why antimicrobial resistance is accelerating globally</li><li>What peptidoglycan is and why it’s essential to bacterial survival</li><li>How targeting cell wall degradation differs from traditional antibiotics</li><li>The strategy behind designing small molecule probes and inhibitors</li><li>How LC-MS is used to analyze peptidoglycan fragments and enzyme activity</li><li>Key analytical challenges: complex mixtures, low-abundance species, and structural similarity</li><li>The importance of sample preparation and reproducibility in LC-MS workflows</li><li>How this research could lead to new antibiotic targets</li></ul><h3><strong>Video Transcription</strong></h3><h4><strong>The Growing Challenge of Antibiotic Resistance</strong></h4><p>According to Reid, antibiotic resistance is increasing across a broad spectrum of bacterial pathogens that were once readily treated with existing therapies. In some cases, clinicians must now rely on only a handful of last-line antibiotics, while resistance is also rising in diseases such as tuberculosis.</p><p>One of the major obstacles is the limited commercial incentive to develop entirely new antibiotics. Because bacterial resistance can emerge relatively quickly after a drug reaches the market, pharmaceutical companies often favor modifying existing antibiotics or combining them with resistance-modifying agents rather than investing in completely new drug classes. As a result, genuinely novel antibacterial targets remain urgently needed.</p><h4><strong>Why the Bacterial Cell Wall Remains an Attractive Target</strong></h4><p>The bacterial cell wall consists primarily of <strong>peptidoglycan</strong>, a highly cross-linked polymer that provides structural integrity, maintains cell shape, and enables bacterial growth and division.</p><p>Many widely used antibiotics—including the β-lactam antibiotics such as penicillins—target enzymes responsible for synthesizing or cross-linking peptidoglycan. These pathways have proven highly successful therapeutic targets because peptidoglycan is unique to bacteria and absent from human cells, minimizing unwanted effects on the host.</p><p>However, most existing antibiotics target only specific stages of peptidoglycan biosynthesis. Reid's research explores another part of this complex pathway: enzymes responsible for degrading the carbohydrate backbone of peptidoglycan during normal bacterial growth and remodeling. These enzymes may represent an underexplored class of antimicrobial targets capable of overcoming existing resistance mechanisms.</p><h4><strong>Targeting the Glycan Backbone</strong></h4><p>A key advantage of focusing on the carbohydrate backbone is its remarkable conservation across both Gram-positive and Gram-negative bacteria.</p><p>Whereas the peptide components of peptidoglycan vary considerably between bacterial species—and frequently acquire mutations associated with antibiotic resistance—the glycan backbone remains highly conserved and tolerates relatively few structural modifications.</p><p>This biological constraint makes enzymes acting on the glycan backbone particularly attractive targets. Reid's group developed a series of <strong>small-molecule probes and inhibitors</strong> designed to recognize enzymes known as <strong>N-acetylglucosaminidases</strong>, which cleave glycosidic bonds within peptidoglycan and generate soluble degradation products.</p><p>Rather than beginning with purified enzymes, the researchers adopted a <strong>top-down phenotypic screening strategy</strong>. Candidate compounds were first evaluated for their ability to inhibit bacterial growth before progressively narrowing the list of potential molecular targets through genetic studies, microscopy, enzyme assays, and biochemical characterization. This iterative workflow ultimately identified compounds capable of inhibiting the desired enzyme class while simultaneously providing valuable chemical probes for studying bacterial cell wall biology.</p><h4><strong>LC-MS as a Central Analytical Tool</strong></h4><p>Liquid chromatography coupled with mass spectrometry plays a central role throughout the project by enabling detailed characterization of structural changes occurring within peptidoglycan after bacterial cells are treated with candidate inhibitors.</p><p>Because intact peptidoglycan is an insoluble polymer, sample preparation begins with isolation of the bacterial cell wall through detergent extraction and differential centrifugation. Additional purification steps remove proteins, nucleic acids, and teichoic acids before enzymatic digestion converts the insoluble material into soluble fragments suitable for chromatographic analysis.</p><p>The resulting digest is analyzed by LC-MS to examine changes in the distribution of peptidoglycan fragments. Chromatographic retention times provide an immediate indication of changes in cross-linking, with larger oligomers such as trimers and tetramers generally eluting later than monomeric fragments. Mass spectrometry then confirms the identity of these fragments by measuring their intact molecular masses, while tandem MS experiments can provide further structural information when required.</p><h4><strong>Sample Preparation Determines Data Quality</strong></h4><p>Reid emphasized that sample preparation represents one of the most critical—and challenging—stages of the entire workflow.</p><p>Because peptidoglycan analysis involves multiple purification, digestion, and reduction steps, even minor procedural differences between samples can introduce analytical artifacts that complicate biological interpretation.</p><p>Maintaining strict consistency throughout sample preparation is therefore essential. Appropriate controls must accompany every experiment, ensuring that observed chromatographic and mass spectrometric differences truly reflect biological responses rather than experimental variability. The complexity of bacterial cell wall composition further increases the analytical challenge, as subtle differences in amino acid substitutions and low-abundance cross-linked species can complicate LC-MS data interpretation.</p><h4><strong>Chemical Biology for Future Antibiotic Discovery</strong></h4><p>Although the identified compounds may eventually serve as starting points for antibiotic development, Reid views them primarily as <strong>chemical biology tools</strong>.</p><p>By selectively inhibiting individual enzymes, these probes allow researchers to investigate previously unexplored aspects of bacterial cell wall remodeling that cannot easily be studied using classical genetic approaches alone.</p><p>The research team is now expanding this strategy by combining their probes with <strong>proximity-labeling proteomics and mass spectrometry</strong>, aiming to map protein interaction networks associated with bacterial cell wall degradation. Such studies may reveal entirely new antibacterial targets and provide deeper insight into the biological processes governing bacterial growth and division.</p><p>Ultimately, the work illustrates how integrating synthetic chemistry, microbiology, molecular biology, and advanced LC-MS can accelerate the search for innovative therapeutic strategies against antimicrobial resistance. As existing antibiotics become progressively less effective, multidisciplinary approaches such as these may play an increasingly important role in identifying the next generation of antibacterial targets.</p><blockquote><p><strong>This text has been automatically transcribed from a video presentation using AI technology. It may contain inaccuracies and is not guaranteed to be 100% correct.</strong></p></blockquote><blockquote><h4><strong>Concentrating on Chromatography Podcast</strong></h4><p>Dive into the frontiers of chromatography, mass spectrometry, and sample preparation with host David Oliva. Each episode features candid conversations with leading researchers, industry innovators, and passionate scientists who are shaping the future of analytical chemistry. From decoding PFAS detection challenges to exploring the latest in AI-assisted liquid chromatography, this show uncovers practical workflows, sustainability breakthroughs, and the real-world impact of separation science. Whether you’re a chromatographer, lab professional, or researcher you'll discover inspiring content!</p><p>You can find <strong>Concentrating on Chromatography Podcast </strong>in podcast apps:</p><ul><li><a target="_blank" rel="noopener noreferrer" href="https://open.spotify.com/show/1wZg67rCjoGju4yhqxI3tL"><strong>Spotify</strong></a></li><li><a target="_blank" rel="noopener noreferrer" href="https://podcasts.apple.com/tz/podcast/concentrating-on-chromatography/id1788429185"><strong>Apple podcast</strong></a></li><li><a target="_blank" rel="noopener noreferrer" href="https://podscan.fm/podcasts/concentrating-on-chromatography"><strong>podscan</strong></a></li></ul><p>and on <a target="_blank" rel="noopener noreferrer" href="https://www.youtube.com/@ChromatographyTalk/videos"><strong>YouTube channel</strong></a></p></blockquote>

 Fighting Antibiotic Resistance with LC-MS: Targeting Bacterial Cell Walls

<p><strong>This study compares a commercial ELISA assay with a validated UHPLC-MS/MS method for determining total ergot alkaloids in wheat and durum samples. In addition to method performance, the authors investigated cross-reactivity and matrix effects to explain differences between immunoassay and chromatographic results.</strong></p><p>While ELISA demonstrated good repeatability and is well suited for high-throughput screening, systematic biases were observed due to compound- and matrix-dependent cross-reactivity. The findings support ELISA as an effective screening tool, whereas UHPLC-MS/MS remains the preferred method for accurate quantification and regulatory confirmation of ergot alkaloids.</p><blockquote><h4><strong>The original article</strong></h4><h4><a target="_blank" rel="noopener noreferrer" href="https://pubs.acs.org/doi/10.1021/acs.jafc.6c05234"><strong>Comparative Evaluation of Total Ergot Alkaloids Determination by ELISA and UHPLC-MS/MS in Wheat: Implications of Cross-Reactivity and Matrix Effects</strong></a></h4><p>Chamali Kodikara*, Dainna Drul, Nandika Bandara, and Sheryl A. Tittlemier*</p><p>J. Agric. Food Chem. 2026, 74, 25, 20015–20027</p><p><a target="_blank" rel="noopener noreferrer" href="https://doi.org/10.1021/acs.jafc.6c05234">https://doi.org/10.1021/acs.jafc.6c05234</a></p><h6>licensed under <a target="_blank" rel="noopener noreferrer" href="https://creativecommons.org/licenses/by/4.0/"><strong>CC-BY 4.0</strong></a></h6></blockquote><p><strong>Selected sections from the article follow. Formats and hyperlinks were adapted from the original.</strong></p><div><p>Ergot alkaloids (EAs) are a structurally diverse group of indole-derived mycotoxins produced primarily by phytopathogenic fungi of the genus <i>Claviceps</i>, most notably <i>Claviceps purpurea</i>. These fungi infect the flowering heads of cereal crops, replacing the developing grain with hardened fungal structures (sclerotia) that contain ergot alkaloids. Cereals such as rye, wheat, barley, oats, and triticale are particularly susceptible, with infection severity strongly influenced by environmental conditions during flowering, including rainfall and humidity. <a href="javascript:void(0);">(1−3)</a> Although improvements in agronomic practices and grain cleaning have reduced the prevalence of visible ergot sclerotia, ergot alkaloids can persist in cereals and cereal-based foods even in the absence of detectable sclerotia. This may occur because sclerotia can fragment during harvesting, handling, cleaning, or milling, producing small particles that are not visually detected but may still contribute ergot alkaloids to grain or processed products, continuing to pose food and feed safety concerns. <a href="javascript:void(0);">(4)</a> <a href="javascript:void(0);">(5)</a> Ergot alkaloids share a common tetracyclic ergoline backbone and occur as structurally related compounds, including ergometrine, ergotamine, ergosine, ergocristine, ergocryptine, and ergocornine, together with their corresponding C-8 epimers. These epimers occur as <i>R-</i> and <i>S-</i>configurations, commonly referred to as -ine and -inine forms, respectively. Epimerization at the C-8 position can occur during fungal metabolism and during sample extraction, storage, and analysis, complicating accurate quantification. <a href="javascript:void(0);">(6,7)</a> Although S-epimers were historically considered biologically inactive, recent evidence indicates that they may contribute to overall exposure and toxicity, supporting their inclusion in analytical and regulatory frameworks. <a href="javascript:void(0);">(3)</a></p><p>Dietary exposure to elevated levels of ergot alkaloids has been associated with a range of adverse health effects, collectively referred to as ergotism, including vasoconstriction, ischemia, neurological disturbances, and hallucinations. <a href="javascript:void(0);">(3)</a> These effects arise primarily from interactions with adrenergic, serotonergic, and dopaminergic receptors. <a href="javascript:void(0);">(8−10)</a> In response, regulatory limits for ergot alkaloids in food have been introduced or revised in several jurisdictions. In the European Union, maximum levels have been established for the sum of 12 ergot alkaloids in cereal milling products under Regulation (EU) 2021/1399, which was subsequently consolidated under Regulation (EU) 2023/915. This sum includes ergometrine, ergosine, ergotamine, ergocornine, ergocryptine, and ergocristine together with their corresponding -inine epimers, increasing the demand for reliable analytical methods to support routine monitoring and enforcement. <a href="javascript:void(0);">(1,11)</a></p><p>Liquid chromatography-tandem mass spectrometry (LC-MS/MS), particularly ultrahigh performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS), is widely regarded as the reference technique for confirmatory determination of ergot alkaloids due to its sensitivity, selectivity, and ability to resolve epimeric forms. Recent methods routinely achieve low microgram-per-kilogram limits of quantification across a range of cereal matrices. <a href="javascript:void(0);">(12−17)</a> However, LC-MS/MS approaches are resource-intensive and typically confined to centralized laboratories, limiting their suitability for high-throughput screening or rapid decision-making in grain-handling and quality-control settings. Consequently, rapid screening methods, most notably enzyme-linked immunosorbent assays (ELISAs), are commonly employed as complementary tools to chromatographic analysis. <a href="javascript:void(0);">(18−21)</a> ELISAs offer reduced analysis time, simplified workflows, and lower per-sample costs, making them attractive for preliminary screening and sample triage. <a href="javascript:void(0);">(22−24)</a></p><p>In contrast to other regulated mycotoxins, however, the commercial availability of ELISAs for ergot alkaloids is limited. While multiple kits have been evaluated, performance has varied substantially across assays and matrices. <a href="javascript:void(0);">(1)</a> At present, the Randox Food Diagnostics Ergot Alkaloids ELISA is the only widely marketed microplate ELISA that quantifies the sum of 12 major ergot alkaloids, comprising ergometrine, ergosine, ergotamine, ergocornine, ergocryptine, and ergocristine together with their corresponding -inine epimers, consistent with the analyte scope used for EU maximum levels for ergot alkaloids in specified cereal products. <a href="javascript:void(0);">(25)</a></p><p>In addition, cereal matrix effects may influence antibody binding and signal generation, potentially resulting in biased quantification relative to LC-MS/MS reference methods. These limitations are particularly relevant given the compositional variability of ergot alkaloids across cereal species, geographic regions, and growing conditions. <a href="javascript:void(0);">(12)</a> Recent reviews emphasize the need for systematic, matrix-specific evaluation of immunoassay performance before such methods can be confidently integrated into regulatory or industrial monitoring programs. <a href="javascript:void(0);">(1,6,11)</a> However, independent evaluations of commercially available ergot alkaloid ELISAs, particularly those benchmarked against validated UHPLC-MS/MS methods using naturally infested wheat samples, remain limited.</p><p>In this context, we hypothesized that measurements of total ergot alkaloids obtained using a commercially available ELISA are influenced by matrix effects and antibody cross-reactivity, resulting in systematic differences compared to UHPLC-MS/MS, which provides compound-specific quantification. To test this, the objective of the present study was to conduct a comprehensive performance evaluation of a commercially available ELISA for the detection of ergot alkaloids in wheat, using UHPLC-MS/MS as a reference method. Specific aims included assessment of repeatability and intermediate precision, extraction efficiency and extract stability, the influence of test portion mass, method accuracy relative to UHPLC-MS/MS, and characterization of antibody cross-reactivity toward individual ergot alkaloids in both solvent and wheat matrix. The results provide data-driven guidance on the appropriate use of ELISA-based screening within tiered analytical strategies for regulatory monitoring and food safety decision-making.</p><h4><strong>2. Materials and Methods</strong></h4><h5><strong>2.3. UHPLC-MS/MS Reference Method</strong></h5><p>UHPLC-MS/MS analysis of ergot alkaloids was performed using the validated Canadian Grain Commission (CGC) method described by Tittlemier et al. (2015). Briefly, ground grain (10 g) was extracted with 50 mL of acetonitrile/aqueous ammonium carbonate (84:16, v/v; 3.03 mM ammonium carbonate) to provide alkaline extraction conditions, then shaken for 30 min and centrifuged at 2450<i>g</i> for 10 min at room temperature. An aliquot of extract (464 μL) was combined with internal standard solution (dihydroergotamine; 20 μL of 4 ng/μL) and aqueous ammonium carbonate (1536 μL), vortex-mixed, and filtered through a 0.45 μm polytetrafluoroethylene syringe filter before injection into the UHPLC-MS/MS system. As in the validated reference method, dihydroergotamine was added after extraction to correct for postextraction analytical variability; extraction recovery was evaluated during method validation. Chromatographic separation was achieved using a reversed-phase <strong>BEH C18 column</strong> (2.1 × 100 mm, 1.7 μm; Waters, Milford, MA, USA) with the autosampler maintained at 15 °C and the column at 30 °C. Mobile phases consisted of (A) 3.03 mM aqueous ammonium carbonate and (B) acetonitrile. The gradient program was as follows: initial, 75% A/25% B; 1.00 min, 60% A/40% B; 5.00 min, 40% A/60% B; 8.00 min, 22% A/78% B; 10.50–11.00 min, 10% A/90% B; and 13.00–16.00 min, 75% A/25% B, with a total run time of 16 min. Detection was performed using positive electrospray ionization in multiple reaction monitoring mode on a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/554"><strong>Waters Xevo triple quadrupole mass spectrometer</strong></a> (Waters, Milford, MA, USA). Under the validated chromatographic conditions, dihydroergotamine did not coelute with ergotamine, ergotaminine, or the other target ergot alkaloids. Identification and quantification were based on retention-time criteria and compound-specific MRM transitions, thereby minimizing potential isotopic or coelution interference. Precautions to minimize epimerization, including controlled handling of standards and extracts and reduced light exposure, were followed as described in the reference method. UHPLC-MS/MS quantification was performed using analyte-specific, matrix-matched calibration curves prepared according to the validated CGC reference method, with calibration levels of 0.046, 0.2, 1, 2, 10, and 20 μg/kg. Dihydroergotamine was used as the internal standard, and concentrations were calculated from internal-standard-normalized analyte responses with method dilution factors applied. Total ergot alkaloid concentration was calculated as the arithmetic sum of all quantified ergot alkaloids and epimers included in the validated method. <a href="javascript:void(0);">(26)</a></p><h4><strong>3. Results and Discussion</strong></h4><h5><strong>3.4. Comparison of ELISA and UHPLC-MS/MS Results</strong></h5><p>For method comparison, UHPLC-MS/MS was used as the chemical reference method, providing compound-specific quantification of individual ergot alkaloids and their epimers. A total ergot alkaloid concentration was calculated as the arithmetic sum of all quantified alkaloids included in the method. This summed value represents a chemically defined total based on compound-specific quantification of each analyte, rather than an immunoreactivity-weighted response governed by antibody affinity. As such, the UHPLC-MS/MS total serves as a chemically defined comparator against which the immunoreactivity-weighted ELISA response can be interpreted. Agreement between the commercial ELISA and the validated UHPLC-MS/MS reference method was evaluated using wheat and durum wheat cargo samples (<i>n</i> = 102) collected over multiple crop years (<a target="_blank" rel="noopener noreferrer" href="https://pubs.acs.org/doi/10.1021/acs.jafc.6c05234#fig5">Figure 5</a>). Linear regression analysis demonstrated a positive association between ELISA and UHPLC-MS/MS results across the full concentration range, indicating that the ELISA was responsive to increasing ergot alkaloid levels. However, the regression slope deviated substantially from unity, with ELISA results systematically lower than UHPLC-MS/MS values, indicating an approximate 2-fold underestimation.&nbsp;</p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/J_Agric_Food_Chem_2026_74_25_20015_20027_Figure_5_Linear_regression_analysis_comparing_total_ergot_alkaloid_concentrations_determined_by_ELISA_and_UHPLC_MS_MS_5d7b5c5c61.jpeg" alt="J. Agric. Food Chem. 2026, 74, 25, 20015–20027: Figure 5. Linear regression analysis comparing total ergot alkaloid concentrations determined by ELISA and UHPLC-MS/MS for wheat and durum wheat cargo samples (n = 102). Each point represents the mean of duplicate measurements (extracted from the same test portion) ELISA determinations for an individual cargo sample, plotted against the corresponding UHPLC-MS/MS result. Statistical analysis was performed using linear regression." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_J_Agric_Food_Chem_2026_74_25_20015_20027_Figure_5_Linear_regression_analysis_comparing_total_ergot_alkaloid_concentrations_determined_by_ELISA_and_UHPLC_MS_MS_5d7b5c5c61.jpeg 197w," sizes="100vw" width="992" height="784"></p><p>Bland-Altman analysis confirmed this proportional bias, showing a consistent negative mean difference between ELISA and UHPLC-MS/MS results across the concentration range (<a target="_blank" rel="noopener noreferrer" href="https://pubs.acs.org/doi/10.1021/acs.jafc.6c05234#fig6">Figure 6</a>). Importantly, the magnitude of disagreement did not increase randomly with concentration; rather, it reflected a systematic offset, indicating that the discrepancy was not driven by analytical imprecision or outliers but by fundamental differences in method response characteristics. This pattern indicates method-level bias rather than random analytical variability, which may arise from combined workflow effects including extraction, matrix response, and immunochemical cross-reactivity. The magnitude of the ELISA-UHPLC-MS/MS bias was substantially greater than the intermediate precision observed for the ELISA quality-control material, supporting the interpretation that the disagreement reflects systematic differences in method response rather than insufficient ELISA repeatability. Such behavior is well documented when immunoassays are compared with chromatographic methods for complex mycotoxin mixtures, including ergot alkaloids and other structurally diverse mycotoxins in cereals and feed. <a href="javascript:void(0);">(28,30,33)</a> These studies consistently report matrix- and profile-dependent bias in ELISA relative to LC-MS/MS, arising from differential antibody cross-reactivity and the summed immunoreactivity response of the assay, because ELISA produces an immunoreactivity-weighted total that depends on antibody affinity toward individual alkaloids and their relative abundance in the sample, whereas UHPLC-MS/MS quantifies compound-specific concentrations of each alkaloid and its epimers using compound-specific transitions and response factors, and total ergot alkaloid concentration is calculated as the sum of these targeted analytes. In contrast, ELISA produces an immunoreactivity-weighted total, determined by antibody affinity toward individual alkaloids and their competitive binding behavior within the assay format. Consequently, agreement between ELISA and UHPLC-MS/MS depends not only on total concentration, but also on the relative distribution of individual alkaloids within each sample. Although differences in extraction solvent composition and pH may influence ergot alkaloid recovery, stability, and epimeric distribution, each method was applied according to its intended analytical workflow. The ELISA was performed using the manufacturer-recommended acidic methanol/water extraction, whereas UHPLC-MS/MS was performed using the validated acetonitrile/aqueous ammonium carbonate extraction procedure. Therefore, the comparison reflects the performance of the complete methods as used in practice, rather than an isolated comparison of extraction solvents. The sequential extraction experiment further showed that repeating the ELISA extraction under the same kit conditions did not substantially increase the ELISA-measured response, supporting the adequacy of the kit extraction within the ELISA workflow while recognizing that it does not directly compare acidic and ammonium carbonate-based extraction conditions. Previous ELISA-LC-MS/MS comparison studies for ergot alkaloids have reported both under- and overestimation depending on assay design and matrix, including differences in alkaloid profile, cereal type, and antibody cross-reactivity. Turner et al. observed substantial overestimation of ergot alkaloids by ELISA relative to HPLC in rye field samples, attributed to strong antibody cross-reactivity toward alkaloids prevalent in rye matrices. Similar profile-dependent bias has also been reported in interlaboratory and method-comparison studies, <a href="javascript:void(0);">(33)</a> demonstrating that this behavior is not unique to a single data set.</p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/J_Agric_Food_Chem_2026_74_25_20015_20027_Figure_6_Bland_Altman_plot_comparing_ELISA_and_UHPLC_MS_MS_measurements_of_total_ergot_alkaloids_1be40cce50.jpeg" alt="J. Agric. Food Chem. 2026, 74, 25, 20015–20027: Figure 6. Bland-Altman plot comparing ELISA and UHPLC-MS/MS measurements of total ergot alkaloids for wheat and durum wheat cargo samples (n = 102)." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_J_Agric_Food_Chem_2026_74_25_20015_20027_Figure_6_Bland_Altman_plot_comparing_ELISA_and_UHPLC_MS_MS_measurements_of_total_ergot_alkaloids_1be40cce50.jpeg 230w," sizes="100vw" width="1437" height="977"></p><p>This study provides a comprehensive evaluation of a commercially available ELISA for total ergot alkaloids, using wheat samples as the primary application matrix, and integrates performance metrics with mechanistic insights into method behavior. While the assay demonstrated strong repeatability, calibration stability, and robustness to procedural factors such as test portion mass and extraction conditions, systematic differences relative to UHPLC-MS/MS were consistently observed. By linking these differences to compound-specific antibody cross-reactivity and matrix-dependent signal modulation, this work clarifies that ELISA-derived results reflect an immunochemically weighted response rather than a stoichiometric total. This distinction is critical for the appropriate interpretation of ELISA data and supports its use within tiered analytical strategies as a rapid screening tool, complemented by UHPLC-MS/MS for confirmatory quantification and regulatory decision-making.</p></div>

Comparative Evaluation of Total Ergot Alkaloids Determination by ELISA and UHPLC-MS/MS in Wheat: Implications of Cross-Reactivity and Matrix Effects

<p><strong>In the week of July 6, the following webinars await you in the field of liquid phase and especially HPLC and LC/MS.</strong></p><blockquote><h4><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinars"><strong>👉 DETAILS AND REGISTRATION IN THE WEBINARS SECTION</strong></a></h4><p><strong>👉 Search and filter in the largest global database</strong> with over <strong>5 500 GC, LC, MS and spectroscopy webinars</strong>.</p></blockquote><h5><strong>1. Waters Corporation: Characterizing Next-Generation Antibody Therapeutics: LC Strategies for Multispecifics, ADCs &amp; AOCs</strong></h5><ul><li>Tu, 7.7.2026 11:00 CEST</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/6317">Registration</a></li></ul><p>In this webinar, we share a pragmatic characterization roadmap using LC-based methods across intact, subunit, and peptide-mapping workflows.</p><h5><strong>2. Separation Science: Advances in Antibiotic Extraction from Water: An Automated SPE Approach</strong></h5><ul><li>We, 8.7.2026 09:00 CEST</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/6319">Registration</a></li></ul><p>Streamline water analysis workflows and gain practical insights for modern environmental monitoring.</p><h5><strong>3. SCIEX: Analysis of per- and polyfluoroalkyl substances (PFAS) in food substances using LC-MS/MS solutions</strong></h5><ul><li>We, 8.7.2026 15:00 CEST</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/6322">Registration</a></li></ul><p>Master PFAS analysis in food safety. Learn how SCIEX LC-MS/MS solutions and QTRAP workflows overcome analytical challenges to ensure accurate and compliant result confirmation.</p><h5><strong>4. Wiley: From liquids to solids: expand what your UV-Vis-NIR Spectrometer can do</strong></h5><ul><li>We, 8.7.2026 16:00 CEST</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/6367">Registration</a></li></ul><p>Discover how the Agilent Cary UV-Vis-NIR spectrometer, together with Specac accessories, enables reliable analysis of liquids, powders, films, pellets, and highly scattering solid samples.</p><h5><strong>5. Thermo Fisher Scientific: Advanced tools to maximize your sample throughput with Chromeleon Software and Vanquish UHPLC</strong></h5><ul><li>Th, 9.7.2026 11:00 CEST</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/6326">Registration</a></li></ul><p>Learn how Chromeleon software and the Vanquish Duo UHPLC system streamline sample analysis, automate routine workflows, increase throughput, and reduce errors through practical demonstrations.</p><h5><strong>6. Bruker: 4D-Metabolomics™ in Action: Real-World Strategies for Deep Discovery and Confident Annotation</strong></h5><ul><li>Th, 9.7.2026 17:00 CEST</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/6328">Registration</a></li></ul><p>Explore how next-generation mass spectrometry technologies, ion mobility, and computational tools are helping researchers move from complex datasets to confident insight.</p><h5><strong>7. Waters Corporation: Cracking the Code on Nitrogen Methods</strong></h5><ul><li>Th, 9.7.2026 17:00 CEST</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/6327">Registration</a></li></ul><p>Learn the fundamentals of nitrogen analysis for wastewater testing. Explore analytical methods, QA/QC, sample handling, compliance reporting, and troubleshooting for accurate environmental monitoring.</p><h5><strong>8. SelectScience: Next-generation metabolomics: From annotation to discovery at scale</strong></h5><ul><li>Th, 9.7.2026 17:00 CEST</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/6281">Registration</a></li></ul><p>Explore how next-generation mass spectrometry, ion mobility, and advanced computational tools accelerate confident small-molecule annotation and discovery in metabolomics.</p><h5><strong>9. SCIEX: LC-MS/HRMS Approaches for Monitoring Organic Micropollutants in Wastewater and Environmental Systems</strong></h5><ul><li>Fr, 10.7.2026 15:00 CEST</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/6370">Registration</a></li></ul><p>Discover how advanced LC-MS/HRMS workflows enable trace-level detection of organic micropollutants, suspect and non-target screening, and environmental monitoring of wastewater systems.</p>

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Thermo Scientific Orbitrap Exploris 480 Mass Spectrometer

Empowering life science researchers to translate each experimental step to a new level of insight with built-in workflows and single-cell sensitivity.

<p><strong>Empowering life science researchers to translate each experimental step to a new level of insight with built-in workflows and single-cell sensitivity. Accelerate your path to impactful results with a next-generation ownership experience coupled with an intelligence-driven experimental approach. Achieve a new standard in performance to study trace samples with extraordinary accuracy, certainty, confidence and simplicity. You'll find market leading quantitative performance to solve your most complex challenges with ease, and the proven qualitative performance you expect from Orbitrap- based mass spectrometry.</strong></p><p>From exploratory to targeted quantitation, find the power and reliability to make large scale studies possible, while reducing everyday hassles as you increase productivity. And achieving it all, remarkably, within a compact footprint. An extraordinary instrument, providing extraordinary results; the Thermo Scientific Orbitrap Exploris 480 mass spectrometer.</p><h4><strong>Key benefits of the Orbitrap Exploris 480 mass spectrometer</strong></h4><h5><strong>Next-generation experience</strong></h5><p>Built with next-generation common hardware and software architecture, the guiding principle of ultimate performance and ease of use delivers effortless access to the value contained within your data.</p><h5><strong>High performance, high-throughput insights</strong></h5><p>Adding greater performance and intelligence to market-leading selectivity and spectral quality imparts productivity, depth, and certainty to small and large- scale studies.</p><h5><strong>Robust performance and exceptional serviceability increase uptime</strong></h5><p>By design, exceptional end-to-end robustness minimizes need for performance-related maintenance, increasing uptime and sample throughput.</p><h5><strong>Ease-of-use and intelligence to empower all users</strong></h5><p>Differences in user expertise can be a challenge in many laboratories. Workflows based on intelligent data acquisition methods leverage new levels of instrument performance to deliver high confidence, and high throughput.</p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Thermo_Fisher_Scientific_Ion_path_and_key_components_of_the_Orbitrap_Exploris_480_mass_spectrometer_3f5791378c.webp" alt="Thermo Fisher Scientific: Ion path and key components of the Orbitrap Exploris 480 mass spectrometer" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Thermo_Fisher_Scientific_Ion_path_and_key_components_of_the_Orbitrap_Exploris_480_mass_spectrometer_3f5791378c.webp 245w," sizes="100vw" width="950" height="410"></p><div><h4><strong>How does the Orbitrap Exploris 480 MS enable greater data insights faster?</strong></h4></div><div><p>See what makes the Orbitrap Exploris 480 MS stand out, how it can empower all users to obtain the highest quality results and deepest insights with the least amount of effort; see how our breakthrough is now your breakthrough.</p></div><ul><li><strong>Next-Generation Instrument Design</strong></li><li><strong>Targeted Protein Quantitation Workflows with SureQuant IS</strong></li><li><strong>Advancing Accuracy, and Precision in Quantitative Proteomics through Multiplexing</strong></li><li><strong>Sub 1-ppm Mass Accuracy as Standard</strong></li><li><strong>Improved Performance through Instrument Enhancements</strong></li><li><strong>Intelligence-Driven Instrument Set-Up, Control and Data Acquisition</strong></li><li><strong>Visualize the Orbitrap Exploris Data Acquisition Modes</strong></li></ul><h4><strong>Optional features to maximize performance</strong></h4><p>Maximize performance and flexibility to expand experimental capabilities of the Orbitrap Exploris 480 mass spectrometer with multiple hardware and software options, allowing you to see more, quantify more, deliver more confidence and greater insights with even more productivity.</p><ul><li><strong>Enhance your Experimental Performance with the FAIMS Pro Duo interface</strong></li><li><strong>Comprehensive and confident biotherapeutic characterization</strong></li></ul><div data-uw-styling-context="true"><h4><strong>Data analysis made easy</strong></h4></div><div data-uw-styling-context="true"><p data-uw-styling-context="true">From complex proteomic challenges, quantitation with SureQuant and TMT multiplexing, through to complete biotherapeutic characterization, data analysis doesn’t have to be the bottleneck to accessing deeper insights into your high-quality MS data. Despite the numerous workflows involved in the analysis of proteins, there are flexible analysis tools which allow simple, yet confident, interpretation of your data to streamline your research and facilitate decision making to take your research to the next level.</p></div><ul><li><strong>Flexibly Identify and Quantify Proteins Based on your Workflow</strong></li><li><strong>The Right Path Towards Complete Biotherapeutic Characterization</strong></li><li><strong>Compliance and Enterprise Readiness with Chromeleon CDS Software</strong><br>&nbsp;</li></ul>

Thermo Scientific™ Orbitrap Fusion™ Tribrid™ Mass Spectrometer

Orbitrap Fusion Tribrid MS combines quadrupole, ion trap, and Orbitrap analyzers for ultra-flexible MSⁿ workflows, enabling deep proteomics, PTM analysis, and confident structural elucidation.

<h4><strong>Revolutionary tribrid architecture for comprehensive structural analysis and deep proteome coverage</strong></h4><p>The Orbitrap Fusion Tribrid mass spectrometer combines a quadrupole, a linear ion trap, and an Orbitrap mass analyzer in a unique architecture that delivers unmatched analytical performance. It provides life scientists with advanced MS<sup>n</sup> capabilities, flexibility in fragmentation techniques, and exceptional resolution and speed — ideal for tackling the most complex analytical challenges in proteomics, PTMs, metabolomics, lipidomics, and small molecule research.</p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Pragolab_Thermo_Orbitrap_Fusion_Tribrid_hmotnostni_spektrometr_e93c577ae5/Pragolab-Thermo-Orbitrap-Fusion-Tribrid-hmotnostni-spektrometr.webp" alt="Thermo Scientific™ Orbitrap Fusion™ Tribrid™ Mass Spectrometer" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Pragolab_Thermo_Orbitrap_Fusion_Tribrid_hmotnostni_spektrometr_e93c577ae5/thumbnail_Pragolab-Thermo-Orbitrap-Fusion-Tribrid-hmotnostni-spektrometr.webp 245w," sizes="100vw" width="1914" height="1059"></p><h3><strong>Key Features and Benefits</strong></h3><h4>🔬 <strong>Revolutionary Tribrid Design</strong></h4><ul><li>Combines <strong>quadrupole</strong>, <strong>linear ion trap</strong>, and <strong>Orbitrap</strong> analyzers</li><li>Perform <strong>MSⁿ up to n = 10</strong> with detection in either ion trap or Orbitrap</li><li>Fragmentation options: <strong>CID</strong>, <strong>HCD</strong>, <strong>ETD</strong>, and <strong>EThcD</strong> at any MS stage</li><li><strong>Synchronous Precursor Selection (SPS)</strong>: up to <strong>20 precursors per scan</strong> for MS³</li></ul><h4>⚙️ <strong>High Performance &amp; Resolution</strong></h4><ul><li><strong>Resolving power up to 500,000 FWHM</strong> at m/z 200</li><li><strong>&lt;1 ppm</strong> mass accuracy with EASY-IC™ internal calibration</li><li><strong>Polarity switching in 1.1 s full cycle</strong></li><li><strong>Scan speeds up to 20 Hz</strong> for Orbitrap and ion trap MSn</li></ul><h4>🧪 <strong>Versatility in Applications</strong></h4><ul><li>Analyze <strong>low-abundance</strong>, <strong>high-complexity</strong>, or structurally complex samples</li><li>Confidently resolve <strong>isobaric interferences</strong> and detect <strong>PTMs</strong></li><li>Full compatibility with <strong>intact protein analysis</strong>, glycoproteomics, and TMT SPS-MS³ quantitation workflows</li></ul><h4>🔁 <strong>Efficient Ion Handling</strong></h4><ul><li><strong>EASY-Max NG</strong> or <strong>NanoSpray Flex NG</strong> ion sources</li><li><strong>Active Beam Guide</strong> and <strong>S-Lens</strong> for efficient ion transmission</li><li><strong>Dynamic Scan Management</strong> optimizes scan scheduling and parallelization</li></ul><h4>💡 <strong>Ease of Use</strong></h4><ul><li>Intuitive <strong>Tune and Method Editors</strong></li><li>Method templates for routine and advanced workflows</li><li><strong>Quanfirmation</strong>: HRAM SIM for quantitation + ion trap MS/MS for confirmation</li><li>Automated calibration, AGC, and exclusion logic</li></ul><h4>🧬 <strong>Compatible Software</strong></h4><ul><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1758"><strong>Xcalibur™</strong></a><strong>, </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1782"><strong>Proteome Discoverer™</strong></a><strong>, </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1756"><strong>ProSight™</strong></a><strong>, </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1757"><strong>Mass Frontier™</strong></a><strong>, </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1759"><strong>Compound Discoverer™</strong></a>, and others</li><li>Support for data-dependent, targeted, and deconvolution workflows</li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Pragolab_Thermo_Orbitrap_Fusion_Tribrid_hmotnostni_spektrometr_2_3d7accb56e/Pragolab-Thermo-Orbitrap-Fusion-Tribrid-hmotnostni-spektrometr-2.webp" alt="Thermo Scientific™ Orbitrap Fusion™ Tribrid™ Mass Spectrometer" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Pragolab_Thermo_Orbitrap_Fusion_Tribrid_hmotnostni_spektrometr_2_3d7accb56e/thumbnail_Pragolab-Thermo-Orbitrap-Fusion-Tribrid-hmotnostni-spektrometr-2.webp 245w," sizes="100vw" width="1038" height="622"></p>

Thermo Scientific™ Orbitrap Fusion™ Lumos™ Tribrid™ Mass Spectrometer

The Orbitrap Fusion Lumos Tribrid MS combines quadrupole, ion trap, and Orbitrap analyzers for high-resolution, ultra-sensitive MSn workflows in proteomics, metabolomics, and biopharma.

<h4><strong>The most powerful tribrid MS platform for deep, multidimensional analysis in proteomics, metabolomics, and structural biology</strong></h4><p>The <strong>Orbitrap Fusion Lumos Tribrid MS</strong> is a cutting-edge mass spectrometer that combines a quadrupole, a linear ion trap, and an Orbitrap™ high-resolution accurate-mass analyzer in a single platform. With unmatched sensitivity, selectivity, and versatility, it enables scientists to tackle complex biological questions with confidence—whether in discovery proteomics, targeted quantitation, top-down protein analysis, glycoproteomics, lipidomics, or small molecule research.</p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Pragolab_Thermo_Orbitrap_Fusion_Lumos_Tribrid_hmotnostni_spektrometr_1_39be5478b4/Pragolab-Thermo-Orbitrap-Fusion-Lumos-Tribrid-hmotnostni-spektrometr-1.webp" alt="Thermo Scientific™ Orbitrap Fusion™ Lumos™ Tribrid™ Mass Spectrometer" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Pragolab_Thermo_Orbitrap_Fusion_Lumos_Tribrid_hmotnostni_spektrometr_1_39be5478b4/thumbnail_Pragolab-Thermo-Orbitrap-Fusion-Lumos-Tribrid-hmotnostni-spektrometr-1.webp 245w," sizes="100vw" width="1823" height="1068"></p><h3>🔬 <strong>Unmatched Analytical Performance</strong></h3><ul><li><strong>&gt;500,000 resolution</strong> with Orbitrap Ultra-High Field analyzer</li><li><strong>Sub-ppm mass accuracy</strong> with real-time calibration</li><li><strong>Attomole-level quantification</strong> using PRM and DIA</li><li><strong>High sensitivity MS/MS</strong> with HCD, CID, ETD, EThcD, and new ETD HD fragmentation</li></ul><h3>🔄 <strong>Flexibility Across Workflows</strong></h3><ul><li><strong>Tribrid architecture</strong>: combine quadrupole precursor selection, ion trap MSn, and Orbitrap detection</li><li><strong>DIA, PRM, and SPS-MS3</strong> quantification workflows with isobaric tags (e.g., TMT, TMTpro)</li><li><strong>Top-down intact protein analysis</strong> with high sequence coverage</li><li><strong>Fast polarity switching</strong> and intelligent acquisition for lipidomics</li></ul><h3>🧪 <strong>Ideal for Multi-Omics and PTMs</strong></h3><ul><li>Deep profiling of <strong>glycopeptides</strong>, <strong>phosphoproteins</strong>, and other post-translational modifications</li><li>Resolution of <strong>isobaric isoforms</strong>, methylation patterns, and histone modifications</li><li>Confident structure elucidation of small molecules with fine isotope resolution</li></ul><h3>💡 <strong>Smart Technology for Productivity</strong></h3><ul><li><strong>Advanced Quadrupole Technology</strong> (AQT) for high transmission and selectivity</li><li><strong>Automatic gain control (AGC)</strong> for optimal ion population</li><li><strong>Easy ETD™ source</strong> and <strong>vent-free maintenance</strong> with source plug</li><li><strong>Method templates and visual method editor</strong> for intuitive setup</li></ul><h3>🧬 <strong>Applications</strong></h3><ul><li>Quantitative and discovery proteomics (e.g., large-scale TMT SPS-MS3)</li><li>Structural characterization of antibodies and proteoforms</li><li>Targeted metabolite quantitation and small molecule identification</li><li>Comprehensive glycoproteomics and lipidomics workflows</li></ul><h3>🧷 <strong>Software Compatibility</strong></h3><ul><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1758"><strong>Xcalibur™</strong></a> and <strong>Tune</strong> for control and calibration</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1782"><strong>Proteome Discoverer™</strong></a><strong>, </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1756"><strong>ProSight™</strong></a><strong>, </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/77"><strong>TraceFinder™</strong></a><strong>, </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1757"><strong>Mass Frontier™</strong></a><strong>, </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1759"><strong>Compound Discoverer™</strong></a> for data analysis across omics</li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Pragolab_Thermo_Orbitrap_Fusion_Lumos_Tribrid_hmotnostni_spektrometr_3_7458a4eb26/Pragolab-Thermo-Orbitrap-Fusion-Lumos-Tribrid-hmotnostni-spektrometr-3.webp" alt="Thermo Scientific™ Orbitrap Fusion™ Lumos™ Tribrid™ Mass Spectrometer" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Pragolab_Thermo_Orbitrap_Fusion_Lumos_Tribrid_hmotnostni_spektrometr_3_7458a4eb26/thumbnail_Pragolab-Thermo-Orbitrap-Fusion-Lumos-Tribrid-hmotnostni-spektrometr-3.webp 245w," sizes="100vw" width="2578" height="1296"></p>

Products focused on LC/MS/MS from Thermo Fisher Scientific - page 3

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