<div>Purifying the target protein is crucial for investigating its specific properties and interactions with biomolecules like DNA, RNA, and other proteins. A range of purification strategies is available, and the optimal approach depends on factors such as the required scale, throughput, and downstream application. In many cases, the process must be fine-tuned through empirical optimization to achieve the desired purity and yield.</div><div><div><div><div><h3>What is Protein Purification?</h3>Protein purification is a critical process in biotechnology and life sciences, aimed at isolating a specific protein from complex biological mixtures such as cell lysates or tissue extracts. The goal is to obtain a highly purified protein sample, free from contaminants like other proteins, nucleic acids, and lipids, to enable accurate analysis of its structure, function, and interactions.This process is especially vital in biotechnological areas inside immunology and microbiology. In immunology, it underpins the production of vaccines, therapeutic antibodies, and diagnostic assays by enabling the isolation of key immune proteins and antibodies. It also supports the investigation of cell signaling and immune response mechanisms. In microbiology, purified proteins are essential for studying pathogens, identifying disease-related antigens, and driving drug discovery efforts. Ultimately, protein purification is fundamental to the development of targeted vaccines, innovative diagnostics, and effective therapeutic strategies.</div></div></div></div><div><div><div><div><h3>Types of Protein Purification Methods</h3></div></div></div></div><div>There are several types of protein purification methods, each chosen based on the protein’s characteristics such as size, charge, solubility, and binding affinity. Below are some commonly used methods:</div><div><div><div><div><h4>Non-Chromatographic Approaches</h4></div></div><div>Non-chromatographic approaches use simpler physicochemical principles—such as solubility, molecular size, or diffusional properties—to separate and purify proteins. These techniques are often employed as initial purification or concentration steps before further refinement.<h5>Precipitation Techniques</h5>Protein precipitation is a widely used approach in purification workflows, typically relying on one of three main methods:<ul><li>Organic solvent precipitation involves adding solvents like ethanol or acetone to lower the solution's dielectric constant. This promotes protein–protein interactions, reducing solubility and leading to aggregation and precipitation.</li><li>Isoelectric point precipitation exploits the pH at which a protein carries no net charge. At this point, solubility is minimized, causing the protein to precipitate from solution.</li><li>Nonionic polymer precipitation uses polymers such as dextran, polyethylene glycol (PEG), or NPEP to alter the solution’s physical properties. These agents create crowding effects that promote protein aggregation without directly binding to the proteins.</li></ul><h5>Dialysis</h5>Dialysis involves placing a protein solution inside a semi-permeable membrane that allows water and ions to pass while the protein remains inside, facilitating buffer exchange.This method is commonly used during protein expression and purification workflows for buffer exchange, desalting, and removal of unwanted small molecules such as imidazole, urea, or ammonium sulfate. The process helps maintain protein stability and activity by gradually equilibrating the sample with the desired buffer conditions.<h5>Immunocapture (Magnetic Bead-Based Purification</h5>Immunocapture uses antibodies immobilized on solid supports like magnetic beads to selectively bind and isolate a target protein. This method is highly specific and particularly useful for low-abundance targets or small-scale prep, such as:<ul><li>Pulling down proteins from complex mixtures</li><li>Capturing tagged recombinant proteins (e.g., FLAG, HA, or His tags)</li><li>Enrichment before western blotting, MS, or ELISA</li></ul>Magnetic beads offer speed, ease of use, and minimal sample loss, making them ideal for lab-scale workflows.<a target="_blank" rel="noopener noreferrer" href="https://www.phenomenex.com/products/biozenspe-sample-preparation-products/biozen-magbeads">Biozen MagBeads</a> provide a fast and efficient method for purifying, isolating, and cleaning up monoclonal antibodies (mAbs), proteins, and peptides. These paramagnetic beads feature a streptavidin-coated surface, enabling high-affinity binding in immuno-based workflows. Compared to traditional sample preparation techniques, magnetic bead-based immunocapture offers greater speed and consistency by delivering high binding capacity with uniform particle size, ensuring accurate and reproducible results in significantly less time.<h5>Centrifugation</h5>Centrifugation of crude protein mixtures is an essential step in the investigation of protein purification and aggregation. Outlined below are the various types of centrifugations employed in protein purification.Preparative Ultracentrifugation Ultracentrifugation is an effective method of purifying proteins from aggregates based on their size, shape and density. Centrifugation, widely recognized as a robust and versatile method, is commonly used to separate complex mixtures of cellular components. Ultracentrifuges separate small particles by spinning at high speeds, utilizing centrifugal force based on size and density.Differential Centrifugation Differential centrifugation operates by separating components based on differences in size and density, using varying speeds of centrifugation to achieve this. Its resolving power ranges from low to medium. A key advantage of this technique is its convenience for processing multiple samples simultaneously, making it effective for purifying crude samples. Additionally, it preserves cell viability and maintains the natural distribution of hematopoietic cell populations.Rate-zonal Gradient Centrifugation This technique separates components based on their sedimentation rate within a medium that contains a density gradient. It offers medium resolving power and is particularly effective for separating multiple components in complex mixtures with high precision. It is also considered a cost-effective and widely used technique in molecular biology and biochemistry.Isopycnic Centrifugation (Equilibrium Centrifugation) Isopycnic centrifugation is an advanced protein purification method that relies on the principle of separating particles according to their intrinsic buoyant density, irrespective of their size or morphology. During the process, proteins migrate through a density gradient until they equilibrate at a position where their density matches that of the surrounding medium, resulting in precise separation without further sedimentation.</div></div></div><div><div><div><div><h4>Chromatographic Approaches</h4>Chromatography is a common method of protein purification and is based on the separation of molecules in a mixture as they move through a stationary phase and a mobile phase. Outlined below are the various types of chromatography employed in protein purification.<h5>Ion Exchange Chromatography</h5><a target="_blank" rel="noopener noreferrer" href="https://www.phenomenex.com/techniques/hplc-ion-exchange">Ion exchange chromatography</a> relies on the interaction between a charged stationary phase and oppositely charged analytes, with cation exchange chromatography attracting positively charged molecules to a negatively charged stationary phase. Successful application of this technique requires careful selection of the appropriate ion exchange column, buffer pH, and salt concentration. This approach is often used in the biopharmaceutical industries as mAb polishing steps, viral clearance and charge variant analyses as part of the product quality control.<a target="_blank" rel="noopener noreferrer" href="https://www.phenomenex.com/">Phenomenex</a> offers a range of protein purification columns. Their ion exchange chromatography solutions include:&nbsp;<ul><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1526">Luna SCX </a>for strong selectivity of positively charged compounds, such as peptides</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1540">Biozen WCX</a> for efficient separation of proteins such as monoclonal antibodies charged variants</li><li><a target="_blank" rel="noopener noreferrer" href="https://www.phenomenex.com/part/00g-3151-e0">PhenoSphere SAX</a>, a strong anion-exchanger with fully porous 80Å particles, particularly useful for small molecules such as nucleotides</li></ul><h5>Size Exclusion Chromatography</h5><a target="_blank" rel="noopener noreferrer" href="https://www.phenomenex.com/techniques/hplc-sec">Size exclusion chromatography (SEC)</a> separates molecules based on their size, with larger analytes eluting first due to their exclusion from the pores in the column packing material, while smaller molecules are able to enter the pores, facing a longer travelling path, and hence, eluting later. This method can be categorized into aqueous SEC (gel filtration chromatography, GFC) for analyzing biomolecules such as proteins and antibodies. This type of chromatography is widely used as an analytical tool for quality control in the biopharmaceutical industry and can also be an approach of choice for polishing steps.Phenomenex offers a range of Biozen columns for SEC separations. In the case of mAbs, <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1534">Biozen dSEC-2</a> and<a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1537"> Biozen dSEC-7 </a>columns are the top choices for monoclonal antibody (mAb) analysis, delivering high-resolution separation of monomers, aggregates, and fragments formed during mAb manufacturing, processing, or storage. The dSEC-7 column, featuring a larger pore size and optimized particle design, extends the dynamic range to accommodate larger biomolecules and complex biotherapeutics.For more general applications, <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1533">Biozen SEC-2</a> and <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1532">Biozen SEC-3</a> phases cover different MW ranges for high-efficiency separation of biomolecules (e.g., proteins and antibodies). Additionally, the BioSep and Yarra lines become top choices when looking for preparative scale columns.<h5>Reversed-Phase Chromatography</h5><a target="_blank" rel="noopener noreferrer" href="https://www.phenomenex.com/techniques/hplc-reversed-phase">Reversed-phase chromatography</a>, the most common mode in high-performance liquid chromatography/ ultra-high-performance liquid chromatography (HPLC/UHPLC), separates compounds based on hydrophobic and van der Waals interactions using alkyl (C18, C8, C4) stationary phases or other hydrophobic polymers. For large biomolecules such as proteins and peptides, <a target="_blank" rel="noopener noreferrer" href="https://www.phenomenex.com/resources/blog/wider-pore-biozen-column">wide-pore</a> (typically 300 Å) C4, C8 and C18 columns are ideal, as the larger pore size allows better accessibility of the protein molecules to the stationary phase surface, improving mass transfer and peak shape during separation.The Biozen line from Phenomenex offers a wide range of HPLC/UHPLC columns for the analysis of biomolecules.These include phases optimized for high-resolution analysis of intact proteins (Native-RP, Widepore C4, Intact XB-C8) and peptides (Peptide PS-C18, Peptide XB-C18 and Peptide Polar-C18). Kinetex could also be an option when looking for more selectivities for the analysis of peptides and small proteins. Additionally, when aiming to preparative scale processes, several of these media are also available in Phenomenex Axia formats to easily up-scale your separations.<h5>Hydroxyapatite Chromatography</h5>Hydroxyapatite chromatography (HAC) is a sophisticated mixed-mode chromatographic technique that exploits the unique properties of hydroxyapatite (Ca10(PO4)6(OH)2), utilizing both cation-exchange interactions via its phosphate groups and metal affinity through its calcium sites to achieve highly selective protein separations.Renowned for its efficacy in resolving complex biomolecular separations, HAC excels in tasks like aggregate removal from monoclonal antibodies, surpassing the capabilities of traditional single-mode chromatographic methods such as ion-exchange or hydrophobic interaction chromatography.<h5>Hydrophobic Interaction Chromatography</h5>Hydrophobic interaction chromatography (HIC) is a method of protein purification that separates molecules based on their hydrophobicity, using high-salt buffers to promote protein-ligand interactions while maintaining the protein's biological activity. The process involves gradually reducing the salt concentration to elute proteins in order to increase hydrophobicity, making it effective for purifying target molecules and removing impurities or aggregates, making it a valuable approach for intermediate and polishing steps during purification processes.<h5>Affinity Chromatography</h5>Affinity chromatography is a powerful and highly selective technique used to purify biomolecules based on specific, reversible interactions between a target molecule and an immobilized ligand. In many cases, recombinant proteins are engineered with affinity tags—such as His-tags or GST-tags—that facilitate binding to complementary ligands on the chromatography matrix.This method offers the advantage of achieving several thousand-fold purifications in a single step while preserving the biological activity of the protein. It is widely employed for purifying enzymes, antibodies, and antigens, using a variety of ligands including specific antibodies, enzymes, and multifunctional compounds like concanavalin A and triazine dyes.</div></div></div></div>

Exploring Protein Purification Methods: Essential Techniques for Accurate Results

<h3>A Practical Guide to Getting the Most from Syringe Filters: Tips, Tricks, and Common Pitfalls</h3>Syringe filters play a crucial role in chromatography and many other laboratory applications by removing particulate matter before analysis. Proper sample filtration helps protect analytical instruments, extends column lifetime, and improves data quality. However, selecting an unsuitable filter—or using one incorrectly—can introduce contamination, reduce filtration efficiency, or even compromise analytical results. The following recommendations will help you avoid common mistakes and ensure reliable filtration every time.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Avoiding_Common_Syringe_Filter_Mistakes_3_150e8c1c2b.webp" alt="KNAUER: Avoiding Common Syringe Filter Mistakes" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Avoiding_Common_Syringe_Filter_Mistakes_3_150e8c1c2b.webp 133w," sizes="100vw" width="1276" height="1496"><h4>1. Improper Filter Handling</h4>Using excessive force during filtration or attempting to reuse disposable syringe filters can damage the membrane and increase the risk of sample contamination.<ul><li>Solution: Apply slow, even pressure throughout the filtration process and always dispose of single-use filters after one use. If the flow rate decreases noticeably, replace the filter instead of applying additional force.</li></ul><h4>2. Selecting an Inappropriate Pore Size</h4>Using the wrong pore size may either fail to remove unwanted particles or cause the membrane to clog prematurely, reducing filtration efficiency.<ul><li>Solution: A 0.45 µm membrane is generally suitable for routine particulate removal, while a 0.22 µm membrane is recommended for finer filtration and bacterial removal. Samples with higher viscosity may require a pre-filter to prevent rapid clogging. It is also important to consider the intended analytical technique. For example, 0.22 µm filters are typically preferred for UHPLC and LC-MS applications, whereas 0.45 µm filters are often sufficient for conventional HPLC analyses.</li></ul><h4>3. Choosing the Wrong Membrane Material</h4>Different membrane materials exhibit different chemical resistance. Using an incompatible membrane may lead to degradation of the filter, sample contamination, or complete filtration failure.<ul><li>Solution: Always verify solvent compatibility before filtration. PTFE membranes are an excellent choice for organic solvents, while PES and nylon membranes are better suited to aqueous samples. If compatibility is uncertain, perform a small-scale trial before filtering valuable samples.</li><li>Pro Tip: For additional guidance, see our blog article "How to Choose the Right Membrane for your Syringe Filter", which provides a detailed comparison of membrane materials and their recommended applications.</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Avoiding_Common_Syringe_Filter_Mistakes_2_e47021fb11.jpg" alt="KNAUER Sepapure® Syringe Filters are available with multiple membrane materials, each identified by a dedicated color code for quick and reliable selection." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Avoiding_Common_Syringe_Filter_Mistakes_2_e47021fb11.jpg 245w," sizes="100vw" width="1580" height="902"><h4>4. Ignoring Filter Capacity</h4>Using a filter with insufficient capacity can result in rapid membrane blockage, increasing filtration time and reducing laboratory productivity.<ul><li>Solution: When filtering larger sample volumes, choose high-capacity syringe filters or introduce a pre-filtration step. Never force liquid through a clogged membrane, as doing so may damage the filter and negatively affect sample quality.</li></ul><h4>5. Overlooking Sample Adsorption</h4>Certain membrane materials may adsorb analytes of interest, leading to lower recoveries and potentially affecting quantitative results.<ul><li>Solution: For protein samples or other adsorption-sensitive analytes, select membranes with low binding characteristics, such as nylon or glass fiber, whenever appropriate. Performing a small recovery test before routine analysis can help identify potential analyte losses.</li></ul><h4>6. Failing to Optimize Filtration Performance</h4>Slow filtration is more than just an inconvenience—it often indicates that the filtration conditions are not optimal.<ul><li>Solution: If filtration becomes unusually slow, investigate possible causes such as membrane clogging, excessive sample viscosity, or an unnecessarily small pore size. Depending on the sample, dilution or sequential filtration using progressively finer filters may significantly improve performance.</li></ul><h4>Final Thoughts</h4>Selecting the appropriate syringe filter is a straightforward process, but overlooking small details can have a significant impact on analytical performance. Choosing the correct membrane material, pore size, and filtration strategy helps improve laboratory efficiency, protects chromatography systems and columns, and contributes to accurate, reproducible analytical results.With the right syringe filter and proper filtration technique, you can minimize sample preparation problems and maximize confidence in every analysis.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Avoiding_Common_Syringe_Filter_Mistakes_4_983a15e289.jpg" alt="KNAUER: Sepapure® Syringe Filter Selection" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Avoiding_Common_Syringe_Filter_Mistakes_4_983a15e289.jpg 245w," sizes="100vw" width="1160" height="627">Happy filtering!

Avoiding Common Syringe Filter Mistakes

News from LabRulezLCMS Library - Week 27, 2026

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Everything from the world of analytical chemistry in one place. We connect people in solving their problems. At Labrulez you will find all the necessary information easily, quickly and clearly. Stop searching and start finding.

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Phenomenex is a global technology leader committed to developing novel analytical chemistry solutions that solve the separation and purification challenges of researchers in industrial, government and academic laboratories.

Phenomenex

Based in Berlin, KNAUER is a medium-sized, owner-managed company that has been serving the sciences since 1962. We develop and manufacture scientific instruments of superior quality for liquid chromatography, including: Analytical HPLC/UHPLC, Preparative HPLC, Fast protein liquid chromatography (FPLC), Multi-column chromatography/Simulated moving bed (SMB) chromatography. High pressure dosing, in-line detection systems, and osmometry are additional business areas.

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Agilent Technologies

Business Review Webinars

Reversed phase is the most common mode of HPLC but if it’s not working for you, don’t despair, there are other options. Perhaps HILIC, Chiral, IEX, GPC/SEC, Affinity, SFC, or HIC might be better suited to your sample type.
In this seminar, we will look at additional chromatographic techniques and discuss:
<ul>
<li>Mechanism of separation
</li>
<li>Column selection and available chemistries
</li>
<li>Typical method and eluent conditions
</li>
</ul>
Becoming familiar with other modes of chromatography and investigating alternative chemistry options can result in an improved separation or provide additional information for your sample.
<blockquote>
Presenter: Jean Lane (Application Engineer, Agilent Technologies, Inc.)
</blockquote>
<blockquote>
Jean has been employed with Agilent since Nov 2010 in technical and applications support and has a primary role supporting Agilent’s full line of LC columns and consumables, which also includes support for Agilent’s GPC/SEC product line. She came to Agilent thru acquisition of Varian and the former Polymer Laboratories. Jean has 20 plus years of technical and applications support for both HPLC and GPC consumable products. In addition to practical chromatography experience in the laboratory, she has had many, many years in her technical role involving customer support providing assistance and troubleshooting for customer chromatography issues.
</blockquote>

Agilent Technologies: There is More to HPLC Than Reversed Phase; Exploring Other Options

There is More to HPLC Than Reversed Phase; Exploring Other Options

There is More to HPLC: Exploring Other Options

Improper LC connections can create bad chromatography.
In this presentation we will discuss:
<ul>
<li>Types of fittings – what is the best option?
</li>
<li>The importance of using the correct capillary
</li>
<li>How improper connections can affect the chromatogram
</li>
<li>How to make a good connection
</li>
</ul>
Something as simple as a connection can potentially cause chromatography problems that may take you a long time to troubleshoot. Careful attention to making a good connection can reduce frustration and save time troubleshooting.
<blockquote>
Presenter: Golnar Javadi (Application Engineer, Agilent Technologies, Inc.)
</blockquote>
<blockquote>
Golnar Javadi is an applications engineer providing online applications assistance and technical support for LC columns and supplies, Sample Preparation products, and Spectroscopy supplies.
</blockquote>
<blockquote>
Before joining Varian and then Agilent, Golnar worked in the pharmaceutical and nutraceutical industries on method development and quality control. She also taught college level chemistry courses for a few years.
</blockquote>
<blockquote>
Golnar then joined Varian as a technical support specialist, providing customers with method development assistance and technical support on Sample Preparation (SPE, SLE, and Filtration), HPLC, GC, ToxiLab and DAT. After the Varian-Agilent merger, Golnar joined Agilent Technologies as an Applications Engineer. She provides technical support to customers on Sample Preparation products, Liquid Chromatography columns, and Spectroscopy supplies.
</blockquote>

Agilent Technologies: All About LC Connections – The Importance of Making a Great One

All About LC Connections – The Importance of Making a Great One

Improper LC connections can create bad chromatography.

LC Connections – The Importance of Making a Great One

Porous particle sizes have decreased to sub-two micron (STM). Separations formerly done on 5 micron particles can now be accomplished on much shorter STM columns in a fraction of the time with equal or better resolution.
For more complex separations, longer columns can generate high plate counts but at much higher pressure. Recently, an alternative particle design, the superficially porous particle (SPP), has developed. The particle concept has been around for over 40 years but the current version consists of a small solid core (e.g., 1.7-µm in diameter) surrounded by a thin porous shell (e.g., 0.5-µm thick) making a 2.7-µm particle diameter.
The efficiency of these SPPs is similar to the STM because of:
<ul>
<li>Consistent spherical shape
</li>
<li>Rapid solute mass transfer
</li>
<li>Narrow particle size distribution
</li>
</ul>
The larger particle size results in a much lower pressure drop when compared to STM particles, making them compatible with older 400 bar instruments as well as UHPLC systems.
This seminar will look at both types of particles and discuss when you might choose one over the other. Examples of pertinent applications will be cited.
<blockquote>
Presenter: Rita Steed (Application Engineer, Agilent Technologies, Inc.)
</blockquote>
<blockquote>
Rita Steed began supporting the LC column line for Agilent in 1999. As a Chromatography Specialist (first with Chromatography, Inc., then directly for Agilent), Rita worked on-site with researchers in Pharmaceutical and other industries presenting technical seminars and assisting researchers with troubleshooting and method development. Rita has over 20 years of Chromatography experience in the Biotechnology, Chemical, and Pharmaceutical industries. She has held positions in Research, Sales, and Technical Service. Rita has earned degrees in Microbiology and Life Sciences/Biochemistry. In her current position, she is an inside Application Engineer supporting LC columns.
</blockquote>

Agilent Technologies: Practical Aspects of High Performance Column Selection in LC: Superficially Porous (SPP) or Sub-two Micron (STM)

Practical Aspects of High Performance Column Selection in LC: Superficially Porous (SPP) or Sub-two Micron (STM)

Both types of particles and discuss when you might choose one over the other.

Superficially Porous (SPP) or Sub-two Micron (STM)

Superficially porous particles offer improved efficiency and performance over similarly sized traditional totally porous particles.
Higher efficiency leads to improved resolution and possible time savings with superficially porous particles, hence their popularity for HPLC analyses. Columns using superficially porous particles are available in a wide variety of particle sizes and stationary phase chemistries. This webinar will go into detail on how a chromatographer can exploit new phase chemistries on superficially porous columns, as well as utilize new chemistries optimized for long life at elevated pH for improved resolution.
What you will learn:
<ul>
<li>How to use the many phase chemistry options available for superficially porous columns as a powerful tool to expedite method development
</li>
<li>How to use pH to manipulate selectivity of ionizable compounds
</li>
<li>Which superficially porous particle column is best for your current laboratory setup
</li>
</ul>
Who should attend:
<ul>
<li>Chromatographers looking to improve their LC method development by taking advantage of selectivity, via new phase chemistries and pH, on superficially porous columns for maximizing resolution and improving results.</li>
</ul>
<blockquote>
Presenter: Anne Mack (Applications Scientist, Agilent Technologies, Inc.)
</blockquote>
<blockquote>
Anne is an application scientist at Agilent Technologies, specializing in small molecule LC columns, ranging from HPLC to UHPLC. Anne has worked at Agilent for almost 10 years. She started as a Field Service Engineer repairing both liquid and gas phase instruments. Since she joined the applications team in 2008, she has supported new LC products in R&amp;D as well as the entirety of the existing small molecule LC column portfolio in Marketing
</blockquote>
<blockquote>
Presenter: Stephen Luke (LC Columns Product Manager, Agilent Technologies, Inc.)
</blockquote>
<blockquote>
Steve became LC Columns Product Manager for Agilent Technologies in January 2014. His long experience with liquid chromatography began during the mid-1980s in the analytical development laboratory of a large pharmaceutical company and has held roles in multiple different functions, including technical support, R&amp;D, and marketing, for leading organizations in the separations science industry.
</blockquote>

Agilent technologies: Maximizing Resolution and Selectivity: Superficially Porous Column Chromatography Options

Maximizing Resolution and Selectivity: Superficially Porous Column Chromatography Options

How a chromatographer can exploit new phase chemistries on superficially porous columns.

Superficially Porous Column Chromatography Options

There are many choices for reversed phase columns.
In this seminar we will look at: The basics of column chemistry including o Silica surface - Chemistry &amp; Physical Properties Bonding Chemistries End-capping Particle size options Totally porous vs. superficially porous columns We will also look at how this fits with our conditions to get good lifetime from our columns.
<blockquote>
Presenter: Rita Steed (Application Engineer, Agilent Technologies, Inc.)
</blockquote>
<blockquote>
Rita Steed began supporting the LC column line for Agilent in 1999. As a Chromatography Specialist (first with Chromatography, Inc., then directly for Agilent), Rita worked on-site with researchers in Pharmaceutical and other industries presenting technical seminars and assisting researchers with troubleshooting and method development. Rita has over 20 years of Chromatography experience in the Biotechnology, Chemical, and Pharmaceutical industries. She has held positions in Research, Sales, and Technical Service. Rita has earned degrees in Microbiology and Life Sciences/Biochemistry. In her current position, she is an inside Application Engineer supporting LC columns.
</blockquote>

Agilent Technologies: A Look at Column Choices

A Look at Column Choices

 The basics of column chemistry including o Silica surface.

Problems with improper connections are often mistaken for column issues and are a source of many calls to tech support.
In this presentation we’ll discuss:
<ul>
<li>The different types of fittings, including Agilent’s new A-line fittings
</li>
<li>The importance of tubing
</li>
<li>What happens when you don’t make a good connection
</li>
<li>How to make a proper connection
</li>
</ul>
The function of a fitting is simple but a connection made incorrectly can cause problems with your chromatography. Careful attention to making a good one can reduce frustration and save time troubleshooting.
<blockquote>
Presenter: Mark Powell (Application Engineer, Agilent Technologies, Inc.)
</blockquote>
<blockquote>
Mark Powell is an Applications Engineer located at Agilent’s Little Falls site in Wilmington, DE. Before joining Agilent in 2011, Mark worked in the pharmaceutical industry synthesizing, purifying and analyzing drug candidates. He provides applications assistance and technical support for Agilent’s HPLC columns and consumables.
</blockquote>

Agilent Technologies: Making LC Connections - The Good, The Bad, and the Ugly

Making LC Connections - The Good, The Bad, and the Ugly

Problems with improper connections are often mistaken for column issues and are a source of many calls to tech support.

C18 is the most common column for HPLC but what if it doesn’t work for your sample. Very polar compounds have poor retention in reversed phase and can be challenging to separate.
In this talk, we will discuss changes to your reversed phase conditions that can help. We’ll also look at the benefits of choosing a more polar stationary phase. If reversed phase doesn’t work you may need to explore other types of chromatography, like HILIC or Ion Pair Chromatography.
<blockquote>
Presenter: Mark Powell (Application Engineer, Agilent Technologies, Inc.)
</blockquote>
<blockquote>
Mark Powell is an Applications Engineer located at Agilent’s Little Falls site in Wilmington, DE. Before joining Agilent in 2011, Mark worked in the pharmaceutical industry synthesizing, purifying and analyzing drug candidates. He provides applications assistance and technical support for Agilent’s HPLC columns and consumables.
</blockquote>

Agilent Technologies: Polar Analytes C18 Didn’t Work, Now What

Polar Analytes C18 Didn’t Work, Now What

Changes to your reversed phase conditions that can help.

Whether you’re new to HPLC or have been doing it for years, you may find that things just aren’t going the way you want them to.
There are many pitfalls or “gotchas” that can cause problems. Even the issue of what tubing and fittings are compatible and which are not can be a serious impediment to a successful HPLC separation.
In this talk, we will discuss:
<ul>
<li>The importance of connections
</li>
<li>Mobile phase considerations
</li>
<li>Quality
</li>
<li>Preventing microbial growth
</li>
<li>Sample/solvent compatibility
</li>
<li>Column characteristics; their importance and limitations
</li>
<li>The importance of sample prep
</li>
</ul>
Avoiding these pitfalls can help you achieve long-term, reliable results.
<blockquote>
Presenter: Rita Steed (Application Engineer, Agilent Technologies, Inc.)
</blockquote>
<blockquote>
Rita Steed began supporting the LC column line for Agilent in 1999. As a Chromatography Specialist (first with Chromatography, Inc., then directly for Agilent), Rita worked on-site with researchers in Pharmaceutical and other industries presenting technical seminars and assisting researchers with troubleshooting and method development. Rita has over 20 years of Chromatography experience in the Biotechnology, Chemical, and Pharmaceutical industries. She has held positions in Research, Sales, and Technical Service. Rita has earned degrees in Microbiology and Life Sciences/Biochemistry. In her current position, she is an inside Application Engineer supporting LC columns.
</blockquote>

Agilent Technologies: Avoiding Chromatography Heartbreak - Common LC Pitfalls

Avoiding Chromatography Heartbreak - Common LC Pitfalls

Whether you’re new to HPLC or have been doing it for years, you may find that things just aren’t going the way you want them to.

Choosing the right column and conditions for your sample is a winning strategy.
In this free webinar, we’ll look at the best matchups for your sample, including:
<ul>
<li>Column options: phase, particle size, dimensions
</li>
<li>Conditions: pH, mobile phase
</li>
</ul>
By the end, you’ll have additional tools to help you put together a game plan for a successful separation
<blockquote>
Presenter: Mark Powell (Application Engineer, Agilent Technologies, Inc.)
</blockquote>
<blockquote>
Mark Powell is an Applications Engineer located at Agilent’s Little Falls site in Wilmington, DE. Before joining Agilent in 2011, Mark worked in the pharmaceutical industry synthesizing, purifying and analyzing drug candidates. He provides applications assistance and technical support for Agilent’s HPLC columns and consumables.
</blockquote>

Agilent Technologies: The LC Column Playbook

The LC Column Playbook

<h3>Building a strong foundation for your analytical workflow</h3>
Your results are only as accurate as your reference. Within Pharma QC, whether you are performing dissolution testing or releasing finished goods, having accurate, reliable and fully characterized Reference Materials is critical. This webinar will provide viewers with an overview of key concepts relating to Reference Materials.
With a multitude of suppliers serving the industry – both compendial and commercial – there are myriad choices of Reference Materials available for a quality control laboratory. We will discuss how different quality grades are “fit for purpose,” best practices in Reference Material production, and what to look for in a Reference Material Certificate of Analysis.
We will also discuss a novel concept: fully characterized Certified Reference Materials (CRMs) qualified against compendial standards. These CRMs are designed to replace a laboratories in-house working standards and pair with a laboratory’s primary Reference Standards to form a strong, well-characterized and fully traceable Reference Material foundation.
<h3>Key Learning Objectives</h3>
<ul>
<li>Understand how different Reference Material quality grades are fit-to-purpose</li>
<li>Provide key concepts in Reference Material manufacture and characterization</li>
<li>Detail Certificate of Analysis (CoA) key attributes that will be common across sources</li>
</ul>
<h3>Audience</h3>
<ul>
<li>Laboratory Directors</li>
<li>Pharmaceutical Quality Control (QC) Managers</li>
<li>Pharmaceutical Quality Control (QC) technicians</li>
</ul>
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Presenter: Michael Hurst (Product Manager - Reference Materials)
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LCMS webinars focusing on Consumables - page 39

MilliporeSigma - Reference Materials in Pharma QC