<div><p><strong>High-performance liquid chromatography (HPLC) plays a vital role in analytical science, enabling accurate separation, identification, and quantification of compounds in different sample matrices. Central to this process is the </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products?category=48&amp;manufacturerItems=50"><strong>HPLC column</strong></a><strong>, where analytes are separated based on their interactions with the stationary and mobile phases.</strong></p><p>For consistent performance, HPLC column care is essential. Without proper care and maintenance, columns can suffer from high backpressure, poor resolution, and irreproducible results. These issues not only affect data quality but, if left unaddressed, significantly increase the risk of permanent system damage and costly operational downtime.</p><p>In contrast, regular HPLC column maintenance ensures sharp peaks, stable retention times, and reproducible outcomes. Key practices of HPLC maintenance help extend column life and protect your workflow. Whether you are running routine tests or developing new methods, this guide offers a practical HPLC maintenance checklist to keep your HPLC system performing at its best.</p></div><div><div><div><div><h3><strong>Why HPLC Column Care and Maintenance Matters</strong></h3><p>Regular HPLC column care and maintenance are essential for the consistent performance of the system. Even a minor neglect is sufficient to cause the accumulation of particulates or debris, leading to partial blockages within the HPLC column. This is often accompanied by serious consequences such as high backpressure, reduced separation efficiency, and unreliable retention times.</p><p>The extended troubleshooting can interrupt workflows. The repairs and replacements demand significant investment, highlighting the importance of HPLC column care and maintenance for performance and productivity.</p><p>HPLC’s periodic maintenance ensures reproducible results, sharper resolution, and longer lifespan. Routine practices such as flushing with appropriate solvents, using guard columns, and storing in recommended conditions help prevent contamination and physical damage. These steps are especially critical in high-throughput labs or regulated environments, where consistency and reliability are non-negotiable.</p><p>This HPLC column care guide emphasizes proactive habits that protect your investment and minimize downtime. Whether you're conducting routine assays or complex separations, proper care ensures your HPLC system remains efficient, accurate, and ready for demanding applications.</p></div></div></div></div><div><div><div><div><h3><strong>Best Practices for Handling HPLC Columns</strong></h3></div></div></div></div><div><p>Understanding how to handle HPLC columns correctly is essential for HPLC preventive maintenance and ensuring consistent results. The practices outlined below help ensure optimal column function and reliable chromatographic outcomes.</p><ul><li><strong>Avoid sudden pressure changes:</strong> Abrupt shifts in flow rate or solvent composition can disturb the packed bed, leading to channeling or void formation. Flow rates should be ramped up gradually to equilibrate columns slowly when switching mobile phases.</li><li><strong>Use guard columns and inline filters:</strong> Guard columns and inline filters serve as protective barriers, capturing particulates and sample impurities before they reach the analytical column. This simple precaution significantly lowers the risk of clogging and high backpressure to <a target="_blank" rel="noopener noreferrer" href="https://www.phenomenex.com/resources/knowledge-center/hplc-knowledge-center/how-to-extend-the-lifetime-hplc-columns"><strong>extend the lifetime of the HPLC column</strong></a>, ultimately postponing the need for costly replacements.</li><li><strong>Use high-purity solvents and salts:</strong> Precipitation and chemical degradation of the stationary phase are preventable with the use of high-purity solvents and salts.</li><li><strong>Store columns in appropriate solvents:</strong> HPLC columns must be stored in appropriate solvents, typically organic solvents for <a target="_blank" rel="noopener noreferrer" href="https://www.phenomenex.com/techniques/hplc-reversed-phase"><strong>reversed-phase columns</strong></a>, or manufacturer-recommended buffers for <a target="_blank" rel="noopener noreferrer" href="https://www.phenomenex.com/resources/knowledge-center/hplc-knowledge-center/principles-of-ion-exchange"><strong>ion-exchange</strong></a> types. Allowing a column to dry out can damage the stationary phase, compromising results. To prevent contamination and solvent evaporation, both ends of the column must be securely sealed during storage.</li><li><strong>Respective recommended temperature, pressure, and pH ranges:</strong> Always operate HPLC columns within the specified temperature, pressure, and pH limits. Exceeding these parameters can compromise the stationary phase or damage column hardware, resulting in permanent performance loss and unreliable results.</li></ul><p>Proper HPLC column care prevents costly issues. Simple, consistent practices protect your column, reduce troubleshooting, and ensure reliable results.</p></div><div><div><div><div><h3><strong>How to Clean and Regenerate HPLC Columns</strong></h3></div></div><div><div>Over time, contaminants can accumulate within the HPLC column. Routine cleaning is essential for effective HPLC column maintenance. Cleaning protocols can differ significantly, depending on the column’s chemistry and its intended application.</div><ul><li><strong>Reversed-phase columns:</strong> It is recommended to flush HPLC columns with solvents of gradually increasing organic content. Researchers prefer starting with water and progressing to methanol or acetonitrile. This gradient approach helps remove hydrophobic residues and restore peak sharpness.</li><li><strong>Normal-phase columns:</strong> Cleaning should start with non-polar solvents such as hexane, later followed by a gentle introduction of polar modifiers such as isopropanol. This sequence assists in dislodging retained compounds without stressing the column.</li><li><strong>Ion exchange columns:</strong> It is generally helpful to begin with low-salt buffer washes, then gradually introduce stronger buffers or adjust pH levels, depending on the column type and manufacturer’s guidance. This approach assists in regenerating active sites and supporting consistent separation performance.</li><li><strong>Hydrophilic interaction liquid chromatography (HILIC) columns:</strong> Cleaning typically starts with high-organic solvents such as acetonitrile, then transitions to aqueous phases. This effectively removes polar contaminants while preserving column integrity.</li><li><strong>Size exclusion columns:</strong> These columns are particularly sensitive and should only be cleaned using buffers specifically recommended by the manufacturer. This precaution helps avoid altering pore structures and ensures consistent performance.</li></ul><p>Use of high-purity solvents is highly recommended to prevent introducing new particulates. In some cases, <strong>backflushing</strong> may help remove stubborn blockages, but only if approved by the column manufacturer.</p><p>A successfully regenerated column offers near-original efficiency, peak shape, and retention time. However, replacement may be necessary if performance issues persist even after proper column cleaning. Knowing <strong>how to clean HPLC columns</strong> properly not only protects your investment but also minimizes downtime, ensuring reliable analytical output across applications.</p></div></div></div><div><div><div><div><h3><strong>Storage Guidelines for HPLC Columns</strong></h3></div></div></div></div><div><p>Proper storage plays a crucial role in long-term HPLC column maintenance. Improperly stored columns can degrade over time, requiring costly replacements.</p></div><div><div><div><div><h4><strong>Remove buffer salts</strong></h4></div></div></div></div><div><p>Before storing, it’s helpful to flush out any buffer salts. Residual salts can crystallize inside the column, clog frits, and cause irreversible damage.</p></div><div><div><div><div><h4><strong>Use appropriate storage solvents</strong></h4></div></div></div></div><div><p>Refer to the manufacturer’s recommendations for suitable storage solvents. For example, some manufacturers recommend storing <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products?category=347&amp;manufacturerItems=50"><strong>HILIC columns</strong></a> in a solvent containing 80–90% acetonitrile and buffers containing 5–10 mM ammonium acetate or ammonium formate.</p></div><div><div><div><div><h4><strong>Label the columns</strong></h4></div></div></div></div><div><p>Clearly label each column with the storage solvent used, the date of storage, and any relevant notes. This helps avoid confusion, especially when managing multiple columns in the lab.</p></div><div><div><div><div><h4><strong>Maintain proper storage conditions</strong></h4></div></div></div></div><div><p>Columns should be stored upright in a clean, temperature-controlled environment. Exposure to direct sunlight, humidity, or extreme heat to be avoided to prevent stationary phase degradation.</p></div><div><div><div><div><h3><strong>Common Mistakes to Avoid in HPLC Column Maintenance</strong></h3></div></div></div></div><div><p>Even experienced analysts can unintentionally reduce column lifespan through small oversights. A thoughtful approach to HPLC column maintenance helps preserve performance and minimize costly replacements. This concise HPLC column care guide highlights frequent errors to watch out for.</p><ul><li><strong>Storing columns with buffer salts:</strong> Residual salts can crystallize during storage, clogging columns and giving you high backpressure. Always choose to flush thoroughly without buffer salts before storing.</li><li><strong>Using mobile phases outside the column’s pH range:</strong> Operating beyond the recommended pH range degrades the stationary phase, affecting retention and resolution.</li><li><strong>Incompatible mobile phases and sample diluents:</strong> Mismatched solvents may cause precipitation inside the column, damaging packing material and disrupting flow. Solvent compatibility is non-negotiable.</li><li><strong>Skipping pre-column filtration:</strong> Without a filter, particulates can enter the column, increase backpressure, and risk permanent damage. Inline filtration or using guard columns offers simple protection.</li><li><strong>Ignoring backpressure spikes:</strong> A sudden increase in backpressure often signals a buildup or blockage. Prompt attention helps prevent serious complications.</li><li><strong>Allowing columns to dry out:</strong> Drying can irreversibly damage the stationary phase, compromising separation efficiency. Keep columns properly wetted during handling and storage.</li></ul><p>By avoiding these missteps and following a consistent HPLC maintenance routine, you can ensure reliable results and <a target="_blank" rel="noopener noreferrer" href="https://www.phenomenex.com/resources/knowledge-center/hplc-knowledge-center/how-to-extend-the-lifetime-hplc-columns"><strong>extend column life</strong></a>.</p></div>

Maintenance and Care of HPLC Columns

<h3><strong>Taming Tricky Liquids: A Deep Dive into Advanced Pipetting</strong></h3><p><strong>If you've ever struggled with pipetting thick gels, foamy detergents, or fast-evaporating solvents, you already know that not all liquids behave the same. Pipetting may seem straightforward, but achieving accurate and reproducible results becomes much more challenging when your sample isn't water. Viscous, volatile, or otherwise demanding liquids can test even the skills of experienced laboratory professionals.</strong></p><p>The good news? With the right techniques and the right tools, you can overcome these challenges and achieve precise, reliable results in every pipetting task.</p><h4><strong>Air and Adhesion: The Hidden Challenges of Accurate Pipetting</strong></h4><p>Most laboratory pipettes use an air-displacement system. They are designed to deliver highly accurate volumes of aqueous solutions, making them ideal for routine laboratory work. Inside the pipette, a small air cushion separates the piston from the liquid. As the plunger is pressed and released, this air cushion expands and contracts, drawing liquid into the tip and dispensing a defined volume.</p><p>This principle works extremely well for water-based samples. However, liquids with different viscosity, volatility, density, or temperature behave differently. Since the air cushion is compressible, changes in air pressure can affect the transferred volume. In addition, some liquids tend to adhere to the inner surface of the pipette tip, further reducing dispensing accuracy.</p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Figure_1_Air_displacement_system_An_air_cushion_separates_the_liquid_from_the_piston_825d9fa003.webp" alt="KNAUER: Figure 1 - Air-displacement system. An air cushion separates the liquid from the piston" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Figure_1_Air_displacement_system_An_air_cushion_separates_the_liquid_from_the_piston_825d9fa003.webp 134w," sizes="100vw" width="1644" height="1920"></p><h4><strong>The Ideal Solution: Positive Displacement Pipettes</strong></h4><p>For non-aqueous and difficult-to-handle liquids, positive displacement pipettes are often the best choice. They are particularly suitable for highly viscous, volatile, infectious, or otherwise challenging samples. Instead of an air cushion, these pipettes use a disposable piston inside the capillary tip that comes into direct contact with the liquid.</p><p>Without an air cushion to compress or expand, sample properties such as density and vapor pressure no longer influence pipetting accuracy. The piston-capillary design also minimizes errors caused by liquid remaining on the inner walls of the tip.</p><p>As a result, positive displacement pipettes provide the highest accuracy and precision when transferring liquids that differ significantly from water.</p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Figure_2_Positive_displacement_system_The_liquid_is_in_direct_contact_with_the_piston_No_air_cushion_is_involved_400e2c6f6a.webp" alt="KNAUER: Figure 2 - Positive-displacement system. The liquid is in direct contact with the piston. No air cushion is involved." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Figure_2_Positive_displacement_system_The_liquid_is_in_direct_contact_with_the_piston_No_air_cushion_is_involved_400e2c6f6a.webp 133w," sizes="100vw" width="767" height="899"></p><h4><strong>When a Positive Displacement Pipette Isn't Available</strong></h4><p>Of course, not every laboratory has access to a positive displacement pipette. If you're working with challenging liquids using a standard air-displacement pipette, several simple adjustments can significantly improve accuracy.</p><p>Switching to reverse pipetting, reducing aspiration and dispensing speed, and recalibrating the pipette using the actual sample instead of water can all make a noticeable difference. Remember that air-displacement pipettes are factory calibrated with water under controlled conditions, so their performance may differ with other liquids.</p><p>Let's take a closer look at how different liquid properties influence pipetting performance—and how to compensate for them when using air-displacement pipettes.</p><h4><strong>Reverse Pipetting – A Simple Technique with Big Benefits</strong></h4><p>When handling viscous, volatile, or foaming liquids with an air-displacement pipette, reverse pipetting is often the most effective way to improve accuracy and reproducibility.</p><p>The technique involves aspirating slightly more liquid than required while dispensing only the desired volume, leaving a small residual amount inside the tip. This extra volume compensates for liquid retained on the tip walls and helps prevent dripping or air bubble formation.</p><p>Although reverse pipetting takes a little practice, it can greatly improve consistency when working with difficult samples.</p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_pipetting_3728b789f4.webp" alt="KNAUER: Reverse Pipetting – The Best Kept Secret for Challenging Liquids" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_pipetting_3728b789f4.webp 245w," sizes="100vw" width="1024" height="286"></p><h4><strong>High- and Low-Density Liquids</strong></h4><ul><li><strong>Examples:</strong> Chloroform, ethanol, sulfuric acid, phosphoric acid</li><li><strong>Challenge:</strong> Liquid density influences the size of the air cushion inside the pipette, which affects the aspirated volume. High-density liquids exert greater force on the air cushion, resulting in less liquid entering the tip than expected.</li><li><strong>Solution:</strong> Recalibrate the pipette using the liquid being transferred.</li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_pipetting_2_3b06909c75.webp" alt="KNAUER: High- and Low-Density Liquids" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_pipetting_2_3b06909c75.webp 121w," sizes="100vw" width="512" height="659"></p><h4><strong>Viscous Liquids</strong></h4><ul><li><strong>Examples:</strong> Glycerol, DMSO, Tween® 20, oils</li><li><strong>Challenge:</strong> Viscous liquids flow slowly. Fast aspiration can introduce air bubbles, while the liquid itself tends to adhere to the tip walls, reducing the dispensed volume.</li><li><strong>Solution:</strong> Use reverse pipetting, aspirate and dispense slowly, and allow additional waiting time after each step. Recalibrate the pipette using the viscous liquid.</li><li><strong>Tip:</strong> Low-retention and wide-bore tips reduce adhesion and allow viscous samples to enter the tip more easily.</li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_pipetting_3_e0a204a5b3.webp" alt="KNAUER: Viscous Liquids" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_pipetting_3_e0a204a5b3.webp 115w," sizes="100vw" width="512" height="694"></p><h4><strong>Volatile Liquids</strong></h4><ul><li><strong>Examples:</strong> Acetone, acetonitrile, ethanol</li><li><strong>Challenge:</strong> Volatile liquids evaporate into the air cushion, increasing internal pressure. This can force liquid out of the tip, leading to droplet formation and sample loss.</li><li><strong>Solution:</strong> Pre-wet the tip at least five times to equilibrate vapor pressure. Use reverse pipetting at a relatively fast speed and avoid unnecessary delays between aspiration and dispensing.</li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_pipetting_4_3d5cf2d665.webp" alt="KNAUER: Volatile Liquids" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_pipetting_4_3d5cf2d665.webp 115w," sizes="100vw" width="512" height="694"></p><h4><strong>Foaming Liquids</strong></h4><ul><li><strong>Examples:</strong> Tween® 20, BSA solutions</li><li><strong>Challenge:</strong> Foam traps air bubbles, reducing the amount of liquid actually transferred.</li><li><strong>Solution:</strong> Pipette slowly using the reverse pipetting technique to minimize bubble formation.</li><li><strong>Tip:</strong> Wide-bore tips reduce foaming, while filter tips protect the pipette from aerosol contamination.</li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_pipetting_5_cdbcb99e19.webp" alt="KNAUER: Foaming Liquids" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_pipetting_5_cdbcb99e19.webp 115w," sizes="100vw" width="512" height="693"></p><h4><strong>Low Surface Tension Liquids</strong></h4><ul><li><strong>Example:</strong> Detergents such as Tween® 20</li><li><strong>Challenge:</strong> Liquids with low surface tension readily spread across plastic surfaces and tend to remain inside standard pipette tips.</li><li><strong>Solution:</strong> Use slow reverse pipetting together with high-quality low-retention tips to minimize liquid adhesion.</li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_pipetting_6_98165f70a1.webp" alt="KNAUER: Low Surface Tension Liquids" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_pipetting_6_98165f70a1.webp 115w," sizes="100vw" width="512" height="694"></p><h4><strong>Infectious, Corrosive, Toxic, or Radioactive Liquids</strong></h4><ul><li><strong>Examples:</strong> Blood samples, DNA/RNA PCR reagents</li><li><strong>Challenge:</strong> Aerosol formation can contaminate the pipette cone, increasing the risk of cross-contamination.</li><li><strong>Solution:</strong> Use standard forward pipetting together with filter tips to prevent contamination. Regularly disinfect the pipette with 70% ethanol or autoclave it when appropriate.</li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_pipetting_7_f20b361eda.webp" alt="KNAUER: Infectious, Corrosive, Toxic, or Radioactive Liquids" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_pipetting_7_f20b361eda.webp 115w," sizes="100vw" width="512" height="693"></p><h4><strong>Liquids with Significant Temperature Differences</strong></h4><ul><li><strong>Examples:</strong> Buffers at 37°C, nucleic acid reagents stored at 4°C or below</li><li><strong>Challenge:</strong> Temperature differences change the volume of the air cushion. Warm liquids expand the air cushion during aspiration, causing under-delivery, while cold liquids contract it, often resulting in over-delivery.</li><li><strong>Solution:</strong> Whenever possible, allow samples and pipettes to reach the same temperature. If this isn't practical, use a fresh tip for every transfer and avoid pre-rinsing the tip.</li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_pipetting_8_21c89b73bb.webp" alt="KNAUER: Liquids with Significant Temperature Differences" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_pipetting_8_21c89b73bb.webp 121w," sizes="100vw" width="512" height="659"></p><h3><strong>The Takeaway</strong></h3><p>Working with challenging liquids doesn't have to compromise your results. Whether you're transferring viscous gels, volatile solvents, or sensitive biological samples, successful pipetting depends on understanding how each liquid interacts with your pipette.</p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Table_1_Recommended_pipetting_techniques_and_best_practices_for_various_liquids_d40299d801.jpg" alt="KNAUER: Table 1 - Recommended pipetting techniques and best practices for various liquids." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Table_1_Recommended_pipetting_techniques_and_best_practices_for_various_liquids_d40299d801.jpg 245w," sizes="100vw" width="1062" height="282"></p><p>By selecting the appropriate technique—such as reverse pipetting, adjusting aspiration speed, pre-wetting tips, choosing low-retention consumables, or using a positive displacement pipette—you can significantly improve accuracy, precision, and reproducibility.</p><p>Mastering pipetting isn't just about technique—it's about understanding the science behind every transfer.</p>

Pipetting Made Easy - Part 2: Challenging Liquids

<p><strong>Our Library never stops expanding. What are the most recent contributions to LabRulezLCMS Library in the week of 27th 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 / ASMS </strong>and<strong> Waters Corporation / ASMS </strong>and application note by<strong> KNAUER!</strong></p><h3><strong>1. Agilent Technologies / ASMS: Down in the Weeds: Automated Fast Screening Workflow for Cannabinoids in Whole Blood Using High Resolution Mass Spectrometry</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/po_blood_cannabinoids_lc_q_tof_mp_694_asms_2026_en_agilent_02f13ac14e.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Distinguishing psychoactive tetrahydrocannabinol (THC) isomers and their metabolites from nonpsychoactive isobars and novel semi-synthetic cannabinoids like hexahydrocannabinol (HHC) is increasingly important to forensic toxicologists, as is the ability to remain up to date with novel compounds that are introduced into the population. This study describes an automated two-step chromatographic workflow that utilizes an initial fast screen method followed by reinjection with specific chromatography to resolve isobars if a suspect is detected in the fast screen on a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=Revident"><strong>Revident LC/Q-TOF</strong></a><strong> </strong>with<strong> </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=1290+infinity+II+lc"><strong>1290 Infinity II LC</strong></a> (Figure 1). This allows for a larger number of samples to be screened more quickly while ensuring chromatographic resolution of isomers for confirmation. The data acquisition mode enables laboratories to interrogate data retrospectively to determine if novel compounds were present in any given sample.</p><h4><strong>Conclusions</strong></h4><ul><li>Clear and concise software workflow for quick screening analysis&nbsp;</li><li>Baseline resolution of THC isomers and interferences in longer method to allow for confident identification&nbsp;</li><li>All Ions acquisition method allows for retrospective data interrogation with fragment analysis</li></ul><h3><strong>2. KNAUER: Scaling up LNP formulation using a Reynolds number-based methodology</strong></h3><ul><li>Application note</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/VPH_0084_knauer_scaling_up_lnp_formulation_using_a_reynolds_number_based_methodology_e05e67ca78.pdf"><strong>Full PDF for download</strong></a></li></ul><p>LNPs have become an essential enabling technology for RNA-based therapeutics and next-generation drug delivery systems. As these applications progress from laboratory research [1,2] to commercial manufacturing, process robustness and scalability become increasingly important. Beyond scalability, consistent product quality is critical.&nbsp;</p><p>In pharmaceutical development, predefined critical quality attributes (CQAs) such as particle size, PDI, encapsulation efficiency and active pharmaceutical ingredient (API) integrity define whether a formulation meets its intended performance and regulatory requirements. These CQAs are established during early formulation development and must be maintained throughout scaleup to ensure product consistency, pharmaceutical efficacy and reproducibility.</p><p>LNPs are typically generated by controlled mixing of a lipid phase in ethanol with an aqueous buffer containing the nucleic acid payload. The rapid solvent exchange initiates lipid self-assembly and determines the resulting nanoparticle characteristics. Because particle formation occurs on timescales comparable to the mixing process, the interplay between hydrodynamics and nanoparticle assembly is a key determinant of final product quality. Even minor variations in mixing conditions can influence particle size distribution, encapsulation efficiency, and overall formulation consistency.&nbsp;</p><p>To establish robust and scalable manufacturing processes, the underlying transport phenomena must be described independently of an operating scale. Dimensionless numbers provide a powerful framework for this purpose. The Reynolds number (Re) characterizes the flow regime and mixing intensity by relating inertial and viscous forces, while the Damköhler number (Da) describes the relationship between the characteristic timescales of mixing and nanoparticle formation. Together, these parameters provide a quantitative basis for understanding and controlling LNP self-assembly.</p><h4><strong>RESULTS&nbsp;</strong></h4><h5><strong>R&amp;D Evaluation and Hydrodynamic Characterization&nbsp;</strong></h5><p>The experimental study was initiated on <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=503&amp;search=IJM+NanoScaler"><strong>KNAUERs IJM NanoScaler</strong></a>, where multiple LNP formulations were developed and characterized under defined process conditions. During this phase, formulation parameters such as lipid composition, flow rate ratio (FRR), and total flow rate (TFR) were systematically varied to establish target CQAs, particularly particle size and PDI. In addition to recording volumetric flow parameters, the corresponding hydrodynamic conditions were quantified by calculating Re for each experimental setting, thereby the prevailing flow regime within the impingement zone.&nbsp;</p><h5><strong>Determination of a Critical Reynolds Number&nbsp;</strong></h5><p>Particle size was evaluated to depend on the Re using the <strong>IJM NanoScaler </strong>(Fig. 1). At Re below 500 particle sizes of 100 nm and larger were observed, indicating insufficient mixing and heterogeneous supersaturation conditions. With increasing Re, particle size decreased until reaching a plateau region at around 52 nm. Above a critical Reynolds number of ~ 750, further increases in flow rates did not significantly affect particle size. This plateau defines the hydrodynamic condition required to achieve minimal particle size for the given formulation.</p><h4><strong>CONCLUSION&nbsp;</strong></h4><p>A critical Reynolds number defining stable particle size and PDI was successfully identified on the R&amp;D-scale IJM NanoScaler. Operating above this threshold ensured consistent nanoparticle characteristics within a defined plateau region.</p><p>By transferring these hydrodynamic conditions to the benchtop system, predefined CQAs were reliably maintained across scales. The data demonstrate reproducible particle size and PDI during scale-up while enabling a substantial increase in throughput. These findings highlight that a Reynolds number driven approach provides a robust and scalable framework for predictable LNP manufacturing using IJM technology.</p><h3><strong>3. Shimadzu / ASMS: High-Throughput Ultra-Short-Chain PFAS Analysis in Drinking Water using LC-MS/MS with Automated Sample Preparation</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/TP_125_High_Throughput_Ultra_Short_Chain_PFAS_Analysis_in_Drinking_Water_using_LC_MSMS_with_Automated_Sample_Preparation_69e7985cce.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Ultra-short- and short-chain PFAS are highly mobile and persistent contaminants that are challenging to measure in drinking water due to high polarity, low molecular weight, poor chromatographic retention, and susceptibility to laboratory background contamination. This study evaluates a high-throughput LC–MS/MS workflow with automated sample preparation for targeted quantification of ultra-short and short-chain PFAS in drinking water. Method performance was assessed using native background, matrix spike recovery, precision, LOQ accuracy, calibration performance, and continuing calibration verification.</p><h4><strong>Method</strong></h4><p>Nine target PFAS were evaluated: TFA, TFMS, PFPrA, PFMOAA, PFEtS, PFBA, PFPrS, TFSI, and PFBS. Samples and calibrants were prepared in acidified 50:50 PFAS-grade water/methanol using PFAS-grade water provided by MilliporeSigma and an automated ePrep sample preparation system equipped with PFAS-free accessories.</p><p>Chromatographic separation was performed on a polar-embedded IBD column with a delay column to improve retention and minimize background interference. Analysis was performed on a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=LCMS-8065XE"><strong>Shimadzu LCMS-8065XE triple quadrupole system</strong></a> within an 8-minute total cycle time with a 10 µL injection volume (figure 2). Drinking-water samples were collected from seven local household sources and spiked at multiple concentration levels. Samples were ran in replicates to evaluate recovery, precision, and method robustness.</p><h4><strong>Conclusion&nbsp;</strong></h4><p>This 8-minute LC–MS/MS method enabled high-throughput analysis of ultrashort- and short-chain PFAS in drinking water. TFA background observed in blanks and unspiked samples emphasized the need for contamination control and matrix-specific background assessment. Practical LOQs were supported at 10 ng/L for most analytes and 50 ng/L for PFMOAA. Recovery and precision were generally acceptable from 100–1000 ng/L, with higher variability at 10 ng/L and for PFMOAA, indicating areas for further optimization. Calibration and CCV performance supported reliable quantification across the reportable range.</p><h3><strong>4. Thermo Fisher Scientific / ASMS: Unlocking the archived proteome: High-throughput, deep FFPE proteome profiling using the Orbitrap Astral Mass spectrometer</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/po_004694_lsms_protein_ffpe_orbitrap_astral_proteomics_po004694_asms2026_na_en_7cd59d64aa.pdf"><strong>Full PDF for download</strong></a></li></ul><p>FFPE tissue specimens are central to clinical pathology and translational cancer research, enabling long-term storage while preserving morphology for histology and molecular analysis. Widely available through biobanks, they support retrospective studies, biomarker discovery, and precision oncology—especially when paired with detailed clinical data. Compared to fresh tissues, FFPE samples are more accessible, scalable, and cost-effective for proteomics.</p><p>However, FFPE proteomics is technically challenging. Formaldehyde-induced crosslinking complicates protein extraction and digestion, while paraffin removal and crosslink reversal add complexity. Traditional workflows are labor-intensive, time-consuming, and prone to variability, limiting reproducibility and throughput. Although recent advances in sample preparation have improved performance, residual contaminants can still affect instrument robustness and scalability.&nbsp;</p><p>To address these challenges, we implemented a streamlined FFPE proteomics workflow that simplifies processing and improves peptide yield and reproducibility. Using FFPE lung tissue and the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/2465"><strong>Thermo Scientific OptiSpray Ion source</strong></a> with the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/836"><strong>Orbitrap Astral Mass spectrometer</strong></a><strong> </strong>across two LC platforms, we achieved sensitive detection of low-abundance peptides from 20– 200 ng inputs. Combined with fast LC methods and acquisition rates of 60, 180, and 500 samples per day, this approach enables deep, scalable profiling. The automated OptiSpray Ion source also reduces instrument contamination, supporting robust, high-throughput analysis of archived clinical tissues for biomarker discovery and translational research.</p><h4><strong>Experimental procedure&nbsp;</strong></h4><h5><strong>Sample preparation&nbsp;</strong></h5><p>Lung tumor and lung normal samples were de-paraffinized using xylene and sequential ethanol washes. The optimized protein extraction protocol and Thermo Scientific EasyPep Mini MS sample prep kit was used to prepare the digest samples from FFPE sections of normal and tumor lung samples. Protein concentration was measured using Thermo Scientific Pierce Rapid Gold BCA Protein Assay kit. The peptides were quantified using the Thermo Scientific Pierce Fluorometric Peptide Assay Kit before LC-MS/MS analysis. For <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=291&amp;search=Evosep+ENO"><strong>Evosep ENO</strong></a>, peptides were loaded on EvoTips .&nbsp;</p><h5><strong>LC-MS analysis&nbsp;</strong></h5><p>Samples were injected onto an Thermo Scientific OptiSpray µPAC Neo Cartridge (P/N OS-UPAC050NAN) and Thermo Scientific OptiSpray µPAC Neo High-Throughput Cartridge (P/N OS-UPAC005CAP) and separated using 20 min (60 SPD) and 6.8 min (180 SPD) gradients, respectively, in direct injection mode at 55°C on a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/398"><strong>Thermo Scientific Vanquish Neo UHPLC System</strong></a>. For 500 SPD (2.3 min) separation, samples were processed on Evosep ENO using an Evosep EV1182 Performance Column at 40 °C. Detailed LC and MS parameters are provided in Tables 1–2.&nbsp;</p><h5><strong>Data analysis Spectronaut® Software&nbsp;</strong></h5><p>The raw DIA files from both the labeled and unlabeled samples were analyzed together using Biognosys Spectronaut® Software in directDIA mode. Exported output files were imported to R Studio (2023.09.0 Build 463) with R (v4.3.1) for downstream data analysis and visualization.</p><h4><strong>Conclusions&nbsp;</strong></h4><ul><li>End-to-end FFPE proteomics is completed in under one day, integrating rapid sample preparation, fast LC–MS, and streamlined data analysis.&nbsp;</li><li>Deep proteome coverage is achieved from FFPE lung tissue, with up to ~8,600 protein groups and &gt;100,000 peptides identified from 200 ng input in 20 min.</li><li>Increasing throughput by up to ~8× (60 to 500 SPD) with moderate loss in depth, retaining ~70% of proteins at 180 SPD and ~45% at 500 SPD.&nbsp;</li><li>Robust quantitative performance is maintained at low sample input (20 ng) and across acquisition speeds, with median CVs ≤10%.&nbsp;</li><li>The workflow enables reliable differential expression and pathway analysis at scale, demonstrating its utility for biological insight into lung tumor and normal tissues.</li></ul><h3><strong>5. Waters Corporation / ASMS: Identification of Novel Per-and-Polyfluoroalkyl Substance (PFAS) Isoform in Textile Using a Multi-Reflecting Time-ofFlight Mass Spectrometer Technology</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/720009415_0b95241dfd.pdf"><strong>Full PDF for download</strong></a></li></ul><div><div><section><div><div><div><div><div><div><p>The poster presents a non-targeted LC-MS/MS workflow for identifying <strong>per- and polyfluoroalkyl substances (PFAS)</strong> in textile materials using high-resolution mass spectrometry. As concerns over PFAS exposure continue to grow, conventional targeted methods often capture only a limited number of compounds. The authors therefore combined <strong>data-dependent acquisition (DDA)</strong> with high-resolution accurate-mass analysis to characterize PFAS profiles more comprehensively, enabling the identification of novel PFAS isoforms and halogenated contaminants with high confidence.</p><p>The analytical workflow was based on a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/191"><strong>Waters ACQUITY Premier UPLC system</strong></a> equipped with a PFAS solution installation kit and an <strong>ACQUITY Premier BEH C18 (2.1 × 50 mm, 1.7 µm) column</strong>, coupled to the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/2433"><strong>Waters Xevo MRT P10 multi-reflecting time-of-flight (MRT) mass spectrometer</strong></a>. Textile reference material was extracted according to <strong>EN 17681-1:2025</strong>, and LC-MS/MS data were acquired in <strong>data-dependent acquisition (Top 5 DDA)</strong> mode. Data processing and compound identification were performed using the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1544"><strong>waters_connect™ Software Platform</strong></a>, combining accurate precursor and fragment masses with isotopic information.</p><p>A key innovation of the study was the implementation of <strong>mass defect-directed DDA</strong>. By creating a database of nearly <strong>19,600 unique PFAS-related compounds</strong> and using their characteristic mass defects to guide precursor selection, the workflow reduced the number of non-specific MS/MS spectra by approximately <strong>25%</strong>. This allowed the instrument to focus on PFAS-like compounds more efficiently, increasing the number of informative fragmentation spectra and improving the identification of previously unknown PFAS species. The approach successfully identified compounds such as <strong>N-ethyl perfluorooctane sulfonamidoethanol (N-EtFOSE)</strong> in textile extracts with excellent mass accuracy (1.1 ppm).</p><p>The authors conclude that <strong>mass defect-driven DDA</strong> significantly enhances non-targeted PFAS screening by reducing unnecessary MS/MS acquisitions while increasing the number of confidently identified PFAS and PFAS-like compounds. They also suggest that this strategy can be extended beyond PFAS analysis to other classes of persistent environmental contaminants, including <strong>polychlorinated biphenyls (PCBs)</strong> and <strong>polybrominated biphenyls (PBBs)</strong>, making it a promising approach for broader environmental and materials analysis.</p></div></div></div></div></div></div></section></div></div>

News from LabRulezLCMS Library - Week 31, 2026

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