<div><p><strong>Two-dimensional liquid chromatography (2D-LC) was introduced to overcome the limited peak capacity of one-dimensional LC (1D-LC), which often fails with highly complex mixtures. By coupling two separation dimensions with distinct retention mechanisms, 2D-LC enhances resolution, selectivity, and analytical depth.</strong></p><p>Its effectiveness stems from orthogonality—the deliberate use of complementary mechanisms, such as reversed-phase and hydrophilic interaction chromatography (<a target="_blank" rel="noopener noreferrer" href="https://www.phenomenex.com/techniques/hplc-hilic-chromatography"><strong>HILIC</strong></a>)—which maximizes peak capacity and enables the separation of compounds unresolved in 1D-LC.</p><p>2D-LC has become integral to diverse applications. In metabolomics and proteomics, it supports large-scale peptide, protein, and metabolite profiling. Pharmaceutical laboratories employ it for impurity profiling, stability studies, and regulatory testing, while environmental and food sciences use it to detect trace pollutants, pesticides, and other contaminants in complex matrices.</p><p>Recent advances, including integration with high-resolution mass spectrometry, automation, and AI-based data processing, have streamlined workflows and reduced labor intensity. Sustainability efforts, such as solvent recycling and miniaturization, further support its adoption. By combining precision, versatility, and scalability, 2D-LC represents a transformative platform for contemporary analytical science.</p></div><div><div><div><div><h3><strong>Principle and Working Mechanism of 2D-LC</strong></h3><p>Two-dimensional liquid chromatography (2D-LC) is based on the principle of orthogonality. This term refers to the degree of independence between two chromatographic separation mechanisms.</p><p>By selecting stationary and<strong> </strong><a target="_blank" rel="noopener noreferrer" href="https://www.phenomenex.com/resources/knowledge-center/hplc-knowledge-center/mobile-phase-in-chromatography"><strong>mobile phases</strong></a> that exploit different chemical properties such as polarity, hydrophobicity, ionic charge, or molecular size, 2D chromatography achieves superior resolving power. This orthogonality ensures that analytes co-eluting in the first dimension can be effectively separated in the second.</p><p>Peak capacity is one of the key performance metrics in 2D liquid chromatography. It approximates the product of the separation capacity of each dimension. For instance, a first dimension with a capacity of 100 and a second of 150 produces an estimated combined peak capacity of 15,000. This exponential gain enables comprehensive profiling of complex mixtures and enhances analytical confidence.</p><p>The workflow of 2D chromatography is systematic and highly automated. It begins with sample injection, after that, analytes are separated in the first column. Modulation or fractionation then transfers portions of this effluent to the second column, either in real-time or via fraction collection.</p><p>The second-dimension separation employs steep gradients and high-pressure systems to achieve rapid resolution. Finally, detection is carried out using UV-Vis, fluorescence, or mass spectrometry, providing both qualitative and quantitative insights.</p><p>Modern 2D-LC platforms incorporate advanced software and automation, which helps them to enable reproducibility, efficient data processing, and minimal operator intervention.</p></div></div></div></div><div><div><div><div><h3><strong>Modes of 2D-LC Operation</strong></h3></div></div></div></div><div><p>Two-dimensional liquid chromatography (2D-LC) can be performed in different modes. Each of these modes is designed to balance resolution, analysis time, and sample coverage. Two majorly used approaches are comprehensive 2D-LC (LC×LC) and heart-cutting 2D-LC (LC–LC). Comprehensive mode provides full sample coverage and the highest resolution; in contrast, heart-cutting mode offers a targeted and efficient approach for specific analytes.</p></div><div><div><div><div><h4><strong>Comprehensive 2D-LC (LC×LC)</strong></h4></div></div></div></div><div><p>In comprehensive 2D-LC, every fraction eluting from the first dimension undergoes further separation in the second dimension. This all-encompassing approach is ideal for untargeted studies, including metabolomics, proteomics, environmental monitoring, and materials science, where sample complexity is extreme.</p><p>LC×LC demands rapid second-dimension separations, generally executed on short sub-2 µm <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products?category=48&amp;manufacturerItems=50">lc-columns</a> under ultrahigh-pressure liquid chromatography (<a target="_blank" rel="noopener noreferrer" href="https://www.phenomenex.com/liquid-chromatography/formats-techniques/lc-formats-uhplc-columns"><strong>UHPLC</strong></a>) conditions. Its strength lies in delivering maximum peak capacity and ensuring no compound is mis-analyzed. However, this method generates large datasets, making advanced data handling, chemometric software, and visualization tools essential for interpretation.</p></div><div><div><div><div><h4><strong>Heart-cutting 2D-LC (LC–LC)</strong></h4></div></div></div></div><div><p>Heart-cutting 2D-LC is a selective mode in which only specific fractions from the first dimension are reanalyzed in the second. This targeted strategy reduces analysis time, conserves solvent, and focuses resolution where it is absolutely required. It is widely applied for impurity profiling, regulatory compliance testing, and focused metabolomics.</p><p>Innovations like multiple heart-cutting (mLC–LC) and selective comprehensive LC×LC (sLC×LC) combine precision with broader analytical coverage. It offers flexible workflows that can adapt to both discovery and routine quality control needs.</p></div><div><div><div><div><h3><strong>When and Why Use Two-Dimensional Liquid Chromatography</strong></h3><p>Two-dimensional liquid chromatography (2D-LC) is mainly chosen when traditional one-dimensional <a target="_blank" rel="noopener noreferrer" href="https://www.phenomenex.com/resources/knowledge-center/hplc-knowledge-center/lc-an-indept-exploration"><strong>liquid chromatography</strong></a> (1D-LC) becomes inadequate to separate complicated mixtures. In most biological, pharmaceutical, and environmental samples, analytes tend to co-elute with similar physicochemical properties, which leads to difficult quantitation and identification.</p><p>The orthogonality of 2D-LC triumphantly overcomes these difficulties through high augmented peak capacity and resolution, providing an information depth that 1D-LC cannot possibly achieve. This strategy is especially useful in the study of intricate sample matrices. Plant extracts, biological fluids, foods, and environmental samples usually contain a vast number of compounds with comparable retention properties numbering in the thousands.</p><p>2D-LC enables the dissection of such chemical complexity to unmask minor constituents that might be biologically active or of environmental interest. It is now critical for untargeted metabolomics, lipidomics, environmental analysis, and natural product profiling, in which extensive coverage is preferred.</p><p>In regulated sectors like pharmaceuticals, two-dimensional chromatography is becoming commonly used to address stringent quality assurance and regulatory considerations. Where 1D-LC is unable to separate key impurities or degradation products, 2D-LC provides a robust platform for impurity profiling, stability testing, and lot-release assays. It reduces the effect of excipients and non-volatile buffer when using MS for impurity identification. The capacity to selectively target and define trace impurities delivers regulatory assurance and product safety.</p><p>Aside from resolution, 2D-LC provides greater sensitivity and quantitation. Simplifying matrix complexity reduces ion suppression in mass spectrometry processes, which enhances signal clarity and detection limits. This feature is essential for exploratory analysis, like finding new biomarkers or monitoring trace contaminants, where accuracy and reproducibility are of utmost importance.</p><p>With increasing analytical needs, 2D-LC is an ideal high-throughput, scalable, and future-proof solution for laboratories handling chemically complex and troublesome samples.</p></div></div></div></div><div><div><div><div><h3><strong>Common Applications of 2D-LC</strong></h3></div></div></div></div><div><p>Two-dimensional liquid chromatography (2D-LC) has evolved into a versatile analytical tool capable of resolving chemically complex samples that challenge conventional methods. By offering dramatically increased peak capacity and orthogonal selectivity, 2D-LC supports diverse industries ranging from pharmaceuticals to materials science. Below are some of its most impactful applications:</p></div><div><div><div><div><h4><strong>Biopharmaceuticals: mAbs, ADCs, and peptides</strong></h4></div></div></div></div><div><p>Biopharmaceutical development relies on 2D-LC to characterize monoclonal antibodies (mAbs), antibody-drug conjugates (ADCs), and therapeutic peptides. It provides high-resolution profiling of charge heterogeneity, glycosylation, and other post-translational modifications, streamlining development and regulatory submissions.</p></div><div><div><div><div><h4><strong>Proteomics and metabolomics</strong></h4></div></div></div></div><div><p>In proteomics, LC×LC enables deep peptide coverage post-digestion, advancing biomarker discovery. In metabolomics, it resolves thousands of metabolites in biological fluids, especially when integrated with high-resolution mass spectrometry.</p></div><div><div><div><div><h4><strong>Natural products and herbal extracts</strong></h4></div></div></div></div><div><p>2D-LC efficiently separates structurally similar phytochemicals, revealing trace bioactives crucial for drug discovery and nutraceuticals.</p></div><div><div><div><div><h4><strong>Food and beverage testing</strong></h4></div></div></div></div><div><p>2D-LC identifies flavor compounds, pesticides, mycotoxins, and additives, supporting quality control and safety standards in food sectors.</p></div><div><div><div><div><h4><strong>Petrochemical and environmental analysis</strong></h4></div></div></div></div><div><p>From petroleum fractions to persistent organic pollutants, LC×LC offers detailed compositional profiling for compliance and environmental protection.</p></div><div><div><div><div><h4><strong>Polymer characterization</strong></h4></div></div></div></div><div><p>2D-LC is used to examine polymer branching, blends, and molecular weight distributions.</p></div><div><div><div><div><h3><strong>Future Trends and Innovations in 2D-LC</strong></h3></div></div></div></div><div><p>Advancements in technology are rapidly transforming two-dimensional liquid chromatography into a more accessible, routine technique:</p><ul><li><strong>Miniaturization:</strong> Micro- and nano-LC systems reduce sample requirements and environmental impact while improving sensitivity and speed.</li><li><strong>Automated modulation:</strong> Innovations like active solvent modulation (ASM) and transfer modulation resolve solvent compatibility issues between dimensions, improving method robustness.</li><li><strong>Integration with mass spectrometry:</strong> LC×LC–MS is increasingly used in omics and pharmaceutical research for comprehensive structural elucidation.</li><li><strong>AI-driven data analysis:</strong> Artificial intelligence and machine learning algorithms simplify data interpretation and reduce analysis times.</li><li><strong>Green analytical chemistry:</strong> Solvent recycling and energy-efficient workflows are making 2D-LC more sustainable, aligning with global eco-conscious initiatives.</li><li><strong>3D chromatography:</strong> Research into three-dimensional chromatography hints at future developments, where even higher resolving power may become available.</li><li><strong>User-friendly software development:</strong> Efforts are underway to create more intuitive and compliant software solutions, making 2D-LC suitable for GMP laboratory environments.</li></ul></div>

2D-LC (Two-dimensional Liquid Chromatography): What, When and Why to Use it

Pipetting Made Easy – Part 1: Best Practice Guide

<p><strong>Our Library never stops expanding. What are the most recent contributions to LabRulezLCMS Library in the week of 20th July 2026? Check out new documents from the field of liquid phase, especially HPLC and LC/MS techniques!</strong></p><blockquote><p><span style="font-size:20px;">👉 </span><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library"><span style="font-size:20px;"><strong>SEARCH THE LARGEST REPOSITORY OF DOCUMENTS ABOUT LCMS AND RELATED TECHNIQUES</strong></span></a></p></blockquote><blockquote><p><strong>👉 Need info about different analytical techniques?</strong> Peek into <a target="_blank" rel="noopener noreferrer" href="https://gcms.labrulez.com/library"><strong>LabRulezGCMS</strong></a> or <a target="_blank" rel="noopener noreferrer" href="https://icpms.labrulez.com/library"><strong>LabRulezICPMS</strong></a> libraries.</p></blockquote><p>This week we bring you posters by <strong>Agilent Technologies / ASMS, Shimadzu / ASMS, Thermo Fisher Scientific / HPLC Symposium </strong>and<strong> Waters Corporation / ASMS </strong>and technical note by<strong> KNAUER!</strong></p><h3><strong>1. Agilent Technologies / ASMS: Comprehensive characterization of oligonucleotides and related impurities using advanced LC/Q-TOF analytical workflows</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/po_oligonucleotide_lc_q_tof_mp_536_asms_2026_en_agilent_b896ffee63.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Therapeutic oligonucleotides are synthetic, short nucleic acid sequences designed to modulate gene expression by targeting RNA or DNA with high sequence specificity. However, their inherent structural heterogeneity makes impurity profiling essential, as during GMP manufacturing, impurities like phosphorothioate to phosphodiester, 5′ and 3′ truncations or extensions, depurination, and abasic species can directly impact both product safety and efficacy. Antisense oligonucleotides (ASO) like Nusinersen (7127.171 g/mol) are indicated for the treatment of spinal muscular atrophy (SMA). Their closely related variants coelute with the main peak, complicating their characterization in conventional one-dimensional LC/MS workflows. To address this challenge, a 2D LC/MS strategy was implemented using the Agilent 1290 Infinity III Bio LC system with multiple heart cutting, coupled to the 6545XT AdvanceBio LC/Q-TOF and MassHunter BioConfirm software. The first-dimension separation uses an ion pairing buffer-based method to achieve higher chromatographic peak resolution, while the second dimension provides MS compatible desalting, enabling enhanced mass spectrometric sensitivity and improved detection of low-level impurities.&nbsp;</p><h4><strong>Instrumentation</strong></h4><p>1290 Infinity III Bio 2D-LC &amp; <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=6545XT"><strong>6545XT Q-TOF</strong></a> includes:&nbsp;</p><ul><li>1290 Infinity III Bio Flexible Pump (G7131A)&nbsp;</li><li>1290 Infinity III Bio High-Speed Pump (G7132A)&nbsp;</li><li>1290 Infinity III Bio Multisampler (G7137A)&nbsp;</li><li>1290 Infinity III Multicolumn Thermostat (G7116B)&nbsp;</li><li>1290 Infinity III Variable Wavelength Detector (G7114B)&nbsp;</li><li>1290 Infinity III Diode Array Detector (G7117B)&nbsp;</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=2D-LC"><strong>2D-LC System</strong></a> with MHC set up</li></ul><h4><strong>Conclusions</strong></h4><ul><li>A 2D-LC/MS workflow enabled effective separation and identification of coeluting impurities.&nbsp;</li><li>HiRes sampling with multi-injection improved resolution and throughput while preserving 1D separation for MScompatible desalting.&nbsp;</li><li>6545XT AdvanceBio LC/Q-TOF delivered accurate mass and MS/MS data for low-abundance impurity detection.&nbsp;</li><li>MassHunter BioConfirm software enabled automated sequence confirmation and impurity identification.&nbsp;</li><li>This integrated platform offers a robust solution for oligonucleotide characterization in therapeutic development.</li></ul><h3><strong>2. KNAUER: Smart Monitoring &amp; Optimization Solutions for SMB Processes: In-Line and Off-Line Tools for SMB Systems</strong></h3><ul><li>Technical note</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/VTN_0048_knauer_in_line_and_off_line_tools_for_smb_systems_50a5f1c1cf.pdf"><strong>Full PDF for download</strong></a></li></ul><p>In liquid chromatography, the purification of target compounds is classically carried out using single column batch chromatography. More efficient approaches include semi – continuous systems with two to three columns and continuous multicolumn systems. The most widely used of the latter is the simulated moving bed (SMB) chromatography which enables the continuous separation of binary or pseudo-binary mixtures. SMB purification is used in various areas such as petrochemical, pharmaceutical or food industry for the production of fine chemicals in different quantities and purities ranging from gram to tons per day. SMB systems vary significantly in size from small lab scale systems operating columns with 8 mm inner diameter (ID) to industrial systems operating columns of onemeter ID or larger.</p><p>Ideally, an SMB process is developed in a smaller lab scale system to reduce solvent and sample consumption compared to development at production scale. Once the separation parameters are established, the SMB process allows for accurate up scaling from lab to production scale. During method development but also production, different tools can be used to monitor and control the process which can be distinguished between in-line and off-line measures. The most crucial parameter, in addition to measuring the pressure, is monitoring the flow rates of the pumps, outlets and/or zones accurately with additional flowmeters. Normally, SMB systems are not equipped with detectors, but UV or RI detectors can be integrated to monitor process stability.&nbsp;</p><p>A standard off-line tool is used to analyse samples taken for one switch or cycle at the raffinate and extract outlets. The resulting chromatograms are used to check, if the separation is working. Further, samples can be taken during the process allowing the depiction of concentration profiles in the different zones. These tools will be described in more detail in the following sections.</p><h4><strong>MATERIAL AND METHODS</strong></h4><ul><li><strong>AZURA Lab SMB system</strong>: SMB, biocompatible, 30 ml/min zone1, 4 ml/min feed, 10 – 130 bar&nbsp;</li><li><strong>AZURA Lab SMB system: </strong>SMB, stainless steel, 30 ml/min zone1, 4 ml/min feed, 10 – 130 bar&nbsp;</li><li><strong>AZURA Pilot SMB system, standard: </strong>SMB, stainless steel, 250 ml/min zone1, 40 ml/min feed, 2 – 100 bar&nbsp;</li><li><strong>AZURA Pilot SMB system, high flow:</strong> SMB, stainless steel, 400 ml/min zone1, 100 ml/min feed, 2 – 60 bar</li><li><strong>Refractive index detector:</strong> <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/2429"><strong>AZURA® RID2.1L HighFlow detector</strong></a> 100 ml/min&nbsp;</li><li><strong>UV detector: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/2428"><strong>AZURA®UVD 2.1S detector</strong></a>, w/o flow cell</li><li><strong>UV detector: </strong>AZURA®UVD2.1S UV detector, fiber optics w/o flow cell</li></ul><h4><strong>CONCLUSION&nbsp;</strong></h4><p>SMB is a continuous separation process that can run for several hours or days without interruption. Therefore, it is important to monitor the process and to be able to adapt to changes during the run. Different tools are available that can be used for process monitoring and method development. These tools can be divided into two main categories: in-line and off-line tools. In-line tools monitor parameters such as pressure and flow rate directly during the run, whereas off-line tools take a sample, which is then analyzed by an HPLC system. The tools described can be added to the KNAUER SMB systems depending on the customer’s needs. The more parameters that are measured, the better the process can be monitored. This is especially helpful during SMB method development.</p><h3><strong>3. Shimadzu / ASMS: Accelerating drug screening in forensic hair analysis by applying high-speed polarity switching in HR LC-MS/MS</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/TP_180_Accelerating_drug_screening_in_forensic_hair_analysis_by_applying_high_speed_polarity_switching_in_HR_LC_MSMS_168f8cc0b2.pdf"><strong>Full PDF for download</strong></a></li></ul><p>In 2023, the Society of Hair Testing (SoHT) published updated consensus guidelines for the forensic analysis of drugs of abuse in hair, defining recommended targets across eight major drug classes: opiates, cocaine, amphetamines, cannabinoids, opioids, methadone, buprenorphine, and ketamine. Legal drivers, accepted use cases, and degree of prescription differ between US and Europe but in both jurisdictions, mass spectrometry is widely applied to support legally defensible conclusions. The challenge for any drug testing in hair samples, whether it is in forensic and safeguarding contexts, workplace testing or a judicial setting is the need to develop methods capable of detecting expanded search panels with the increased prevalence of new psychoactive substances (NPS) and non-medical prescription drug use. The approach applied in this work considered a LC/QTOF analysis using DDAMS/MS with rapid polarity switching with a novel data processing application, Insight Profiler. The software was designed to support a single click approach to automate data processing of complex data sets in non-targeted analysis from feature detection to compound identification.</p><h4><strong>Materials and Methods</strong></h4><h5><strong>Reversed phase LC Separation: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=Nexera+X2"><strong>Nexera X2&nbsp;</strong></a></h5><ul><li>Shim-pack Velox Biphenyl (100 x 2.1 mm, 2.7 µm) column; column temp. 40 ºC, flow rate: 0.3 mL/min, 17 min total analysis time. Methanol:water gradient with 2 mM ammonium formate and 0.002% formic acid&nbsp;</li></ul><h5><strong>High resolution QTOF analysis: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=LCMS-9050"><strong>LCMS-9050&nbsp;</strong></a></h5><ul><li>MS scan m/z 100-1000, 100 msec scan time. Polarity switching time 800 msec.&nbsp;</li><li>MS/MS DDA: 4 scans in positive ion mode (m/z 40-1000, 33 msec), 2 scans in negative ion mode (m/z 40-1050, 33 msec) CE 5-55V (1.998 sec total cycle time).</li></ul><h5><strong>Data processing</strong></h5><p><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=Insight+Profiler"><strong>Insight Profiler</strong></a>, feature detection threshold set to low, compound identification search lists included NIST, HighResNPS, Shimadzu Forensic Toxicology database.</p><h4><strong>Conclusions&nbsp;</strong></h4><ul><li>The targeted DDA-MS/MS polarity switching approach successfully identified a significant number of unknown compounds in the samples not routinely targeted by triple quadrupole analysis.&nbsp;</li><li>The method using DDA-MS/MS was easily configured to support an expanded DoA target panel to meet the needs of forensic toxicology screening.&nbsp;</li><li>Non-targeted analysis using Insight Profiler accelerated data analysis by aligning identified compounds from the same hair donor showing direction of change for pharmaceutical and illicit compounds detected.</li></ul><h3><strong>4. Thermo Fisher Scientific / HPLC Symposium: Enhancing semi-volatile analyte detection in HPLC-CAD using temperature coupling mode for optimized evaporation control</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/po_004766_hplc_vanquish_cad_fatty_acid_quantification_po004766_na_en_4a0868c2cb.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Polysorbates are widely used in biopharmaceutical formulations but can degrade or contain FAs impurities that must be accurately measured to ensure product quality and safety. Charged Aerosol Detection is commonly used for FA analysis, though its performance depends strongly on EvapT. Higher EvapT improves S/N ratios but can reduce sensitivity for semi-volatile compounds like FAs, while lower EvapT has the opposite effect. The <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/2176"><strong>Thermo Scientific Vanquish Charged Aerosol Detector P</strong></a> series introduces a Temperature Coupling Mode that links EvapT with the charging detection module (CDM) temperature, overcoming limitations of earlier systems with fixed CDM temperature of 40 ºC. This coupling improves response and measurement reliability for semi-volatile species, enabling better optimization of FA analysis<sup>1</sup>.</p><h4><strong>Conclusions&nbsp;</strong></h4><ul><li>Advanced temperature control in CAD improved method sensitivity up to ~47% for semi-volatile fatty acids.&nbsp;</li><li>The best overall S/N was achieved at 25.0 °C EvapT with Temperature Coupling Mode enabled.&nbsp;</li><li>The optimized CAD approach supports more sensitive FA impurities and degradants quantification in pharmaceutical and biopharmaceutical formulations.</li></ul><h3><strong>5. Waters Corporation / ASMS: High mass and spatial resolution MS imaging of soft fruit sample cross-sections using desorption electrospray ionization (DESI)</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/720009492_282619e388.pdf"><strong>Full PDF for download</strong></a></li></ul><div><div><section><div><div><div><div><div><div><p>This poster demonstrates the use of <strong>desorption electrospray ionization mass spectrometry imaging (DESI MSI)</strong> for visualizing the spatial distribution of metabolites in high-water-content soft fruits, specifically strawberries and raspberries. The authors developed a simple direct-transfer workflow to overcome the challenges associated with imaging delicate fruit tissues and compared three transfer substrates: plain glass microscope slides, nitrocellulose membranes, and Hamamatsu Poropare plates. The goal was to determine which surface best preserves metabolite localization while enabling high-quality mass spectrometry imaging.</p><p>Fresh fruit cross-sections were transferred to each substrate by direct contact for 10 seconds before analysis using a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=5&amp;search=DESI+XS"><strong>Waters DESI XS ion source</strong></a> coupled to a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/2433"><strong>Waters Xevo™ MRT P10 high-resolution mass spectrometer</strong> </a>operating in negative electrospray ionization mode. MS imaging was performed with a <strong>50 µm pixel size</strong>, using a <strong>95:5 methanol/water</strong> spray solvent, while sucrose (m/z 341.1089) served as the lock mass for accurate mass calibration. High mass resolution enabled tentative assignment of metabolites such as citric acid, asparagine, malic acid, glutamic acid, palmitic acid, stearic acid, and sucrose.</p><p>The comparison of transfer substrates revealed significant differences in metabolite localization and image quality. Glass slides produced the strongest metabolite signals but also exhibited considerable metabolite delocalization during sample transfer, reducing spatial accuracy. Nitrocellulose membranes transferred a broader range of metabolites and provided complementary chemical information, although they also showed greater analyte spreading and suffered physical damage after a single DESI acquisition. In contrast, <strong>Poropare plates</strong> minimized metabolite delocalization and preserved fine structural details within the fruit cross-sections, despite producing somewhat lower signal intensities than glass slides.</p><p>The study concludes that direct transfer combined with DESI MSI is an effective approach for imaging soft fruit tissues that are otherwise difficult to analyze because of their high water content. Among the evaluated substrates, <strong>Hamamatsu Poropare plates</strong> provided the best balance between spatial resolution and preservation of metabolite localization, making them particularly suitable for high-resolution metabolomic imaging, while high mass accuracy facilitated confident tentative metabolite identification.</p></div></div></div></div></div></div></section></div></div>

News from LabRulezLCMS Library - Week 30, 2026

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2D-LC (Two-dimensional Liquid Chromatography): What, When and Why to Use it

News from LabRulezLCMS Library - Week 30, 2026

Pipetting Made Easy – Part 1: Best Practice Guide