There are countless benefits of employing a virtually pulseless pump in FPLC systems. Discover the advantages of detector accuracy. Consistent flow enhances protein purification and ensures the accuracy and quality of your fractions.

In protein purification, precision is everything, therefore quality FPLC components are essential.   From sample injection to fraction collection, every module of your Fast Protein Liquid Chromatography (FPLC) system plays a critical part in ensuring accurate, consistently reproducible results. The FPLC pump module is the heart of the FPLC system. A virtually pulseless FPLC pump module can significantly improve the quality of your purified protein.

Many chromatographers still run their FPLC system with a standard reciprocating pump that produces pressure pulsations. If you are one of them, you may be compromising peak resolution, gradient stability, and UV detector performance. Switch to a virtually pulseless FPLC pumping module and realize higher protein purification yields.

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What are the benefits of a virtually pulseless pump?

A virtually pulseless pump delivers a continuously smooth flow of liquid without fluctuations in flow rate. FPLC systems typically use standard reciprocating piston pumps, which create pressure pulsation or flow pulsation. These pulsations are created due to the switch over from one piston to the next.

Virtually pulseless pumps utilizing independently controlled dual linear drives with proprietary smoothing algorithms create the most consistent flow possible. Steady flow performance offers major benefits for your FPLC system.

1. Improved Gradient Consistency and Reproducibility

The accuracy of your gradient depends on consistent flow rates from the FPLC pump(s) delivering buffer A and buffer B. Pulsations in flow disturb gradient formation causing erratic protein elution profiles, poor reproducibility and inferior resolution.

Virtually pulseless flow ensures precise gradient control, providing cleaner separations and more reproducible retention times. These benefits are crucial for FPLC development and scaling up your FPLC purification method.

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2. Enhanced Detector Stability and Accuracy

    FPLC UV detectors, conductivity and pH monitors require a steady flow of buffer through the flow cell. In systems that have standard pumps there will be fluctuations in the UV detector baseline                         even when no proteins are eluting. These baseline fluctuations can obscure small protein peaks, affect the quantification and produce spurious noise in the chromatogram.

3. Better Fraction Collection Precision

When your FPLC system collects protein fractions based on UV absorbance or volume, pulsating flow can impede accurate fraction collection. The result can cause peak tailing across multiple tubes, loss of protein yield, contaminated or diluted fractions. 

When using a virtually pulseless pump, the pumping module delivers buffer at a constant and reproducible flow rate, which ensures accurate fraction collecting and crisper separations.

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4. Minimal Maintenance Due To Less Mechanical Wear

Repetitive stress on your hardware causes premature failure of system valves, detector flow cells, tubing, fittings and seals.The result of this repetitive stress leads to increased leaks and wear, more frequent valve maintenance and FPLC system downtime.

Virtually pulseless pumps operate more smoothly, minimizing wear on FPLC components and extending the lifespan of your FPLC system.

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  5. Longer Lasting Columns

FPLC pump pulsations over time prematurely compact your chromatography resin. This leads to loss of column resolution and premature failure of the column. Virtually pulseless FPLC pumps operate more consistently preventing pounding on the head of your column and ultimately extending the lifespan of your FPLC column.

6. More Reliable Baselines

A virtually pulseless FPLC pump flow profile provides sharper chromatographic peaks resulting in more stable baselines. 

In FPLC pumping modules, accurate and reproducible flow is critically important. ONLY virtually pulseless pumps deliver that accuracy!

Whether you’re purifying proteins, antibodies, or nucleic acids, a smooth and uninterrupted flow improves every step of your purification. Better detector performance, sharper peaks, cleaner fractions and reduced wear-and-tear on both your FPLC system and column are some of the benefits of a virtually pulseless pump.

Consider custom building a modular FPLC system,  a virtually pulseless pump is one of the most valuable improvements you can make. In FPLC,  precision begins with flow.

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Do you have interest in the benefits of a virtually pulseless pump? If so, we are happy to help you choose the right model based on your flow rate, pressure range, and application needs.

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Gel Filtration Chromatography is also known as Size-Exclusion Chromatography. This technique separates molecules based on their size. It is used to separate both proteins and other molecules. Gel Filtration chromatography is a form of partition chromatography used to separate molecules based on their molecular size. Molecules are partitioned between the mobile and stationary phase. The gel medium is comprised of specific size distribution of spherical beads. The separation takes place when the sample is passed through the beads (the gel medium). Smaller molecules pass through the beads and flows through the column faster than larger molecules. The proteins are eluted from the column in order of decreasing size. Therefore, molecules are separated based on their size as they pass through the column and may be eluted in order of decreasing molecular weight. For this reason this mode of FPLC is sometimes refered to as Molecular-seive chromatography. Gel filtration chromatography is used for protein purification, determining molecular weight and for analyzing protein complexes.

Ion Exchange Chromatography (IEC) is a technique that separates charged molecules (proteins) based on their net charged by binding them to a charge resin with opposite polarity. Charged compounds are absorbed and retained by the ion exchanger with the opposite charge. The neutral or similarly charged compounds pass through the void volume and are eluted from the column. Absorbed compounds are eluted with a salt or pH gradient. It is used to analyze ionic compounds, purify proteins, and separate ions and ionizable molecules. 

Affinity Chromatography is a technique used to purify a specific molecule from a complex mixture based on its affinity for a complementary molecule, this complementary molecule is called a ligand. The ligand is bound to a solid support, the resin (column material). Next, the sample is run in a column and the target molecule (usually protein) binds to the ligand. None of the bound components are washed away leaving only the protein of interest bound to the ligand. In the elution stage, the target protein is released from the ligand.

Affinity Chromatography is used for protein purification, antibody purification,  enzyme purification,  drug discovery and biomarker detection. Reversed-Flow Chromatography is for Affinity Purifications..
Hydrophobic interaction chromatography (HIC) is a technique used to separate and purify molecules (primarily proteins) based on their hydrophobicity. Molecules with more hydrophobic regions bind more strongly to a hydrophobic stationary phase on a chromatography column, allowing for separation based on their relative hydrophobicity. The salt in the buffer reduces the solvation of sample solutes. This separation is typically achieved by gradually decreasing the salt concentration in the mobile phase. High salt concentrations in the mobile phase promote hydrophobic reactions.  Therefore, proteins are usually loaded onto the column in a high salt buffer. This causes less hydrophobic molecules to elute first. To elute bound proteins, the salt concentration is gradually decreased. This weakens the hydrophobic interactions. Proteins can then separate based on their relative hydrophobicity. Hydrophobic Interaction Chromatography is widely used for antibody purification, vaccine development, plasma protein fractionation and drug discovery. Different than other FPLC techniques, HIC operates under relatively mild conditions, this helps to preserve the native structure and function of the proteins. Using HIC, researchers can isolate proteins based on how much of their surface area is exposed as hydrophobic patches, allowing the removal of impurities and aggregates with different hydrophobic properties. HIC is widely used to purify proteins from complex mixtures like cell lysates. 
Reversed Phase Chromatography (RPC) is not commonly used. It is a technique used to separate, analyze, and purify organic compounds. RPC separates compounds based on their hydrophobicity. The stationary phase is nonpolar, while the mobile phase is polar. Reversed phase chromatography is used to separate and purify molecules like proteins, peptides and antibodies. RPC is easy to handle and has high resolution. Reversed phase is used to analyze peptides and proteins, separate drugs and metabolities, study environmental contaminants and to purify biological molecules.

There are one step purifications and multi step purifications depending on what matrix your protein is in and how complex the separation is. In the simplest of cases, there is a huge difference in the molecular weight in the protein of interest vs other proteins. In that case, gel filtration or sizing might be the purification method if choice. In most cases, a combination of two or more techniques (Gel filtration/size exclusion chromatography, ion exchange chromatography, affinity chromatography and reversed phase chromatography) are needed to separate and highly purify your protein of interest.

Call or email Icon Scientific, Inc. to figure out what best fits your needs.

301-330-ICON (4266)

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Benefits of Remote Flow Cells

Traditionally, UV detectors in liquid chromatography systems have employed a flow cell positioned on the front of the UV detector. However, the introduction of remote flow cells has opened up new possibilities. Placing the flow cell at the end of the column minimizes dead volume between the UV detector and the column, resulting in reduced peak broadening. This innovation is especially beneficial in fast protein liquid chromatography (FPLC),  preparative liquid chromatography, and on liquid chromatography process skids.

In certain scenarios, having a remote flow cell becomes imperative. For instance, in processes involving flow reaction chemistry systems, the advantages of remote sensing are particularly pronounced. The use of fiber optics in UV detectors with remote flow cells further enhances their applicability and efficiency.

Applications of Remote Flow Cells

The primary applications benefiting from UV detectors with remote flow cells are diverse, ranging from fast protein liquid chromatography to preparative liquid chromatography and liquid chromatography process skids. Additionally, these detectors find utility in the separation and purification of PET isotopes, where the flow cell can be strategically placed within a radioactive environment while the UV detector remains outside, ensuring safety and precision.

Challenges and Solutions

One of the key challenges in implementing UV detectors with remote flow cells is the need for special fiber optic cables. Standard fiber optic cables are unsuitable for the UV region. The length of these cables is crucial, with applications sometimes requiring cables as long as 10 meters. However, three-to-four-meter fiber optic cables are commonly used in most scenarios.
The sizing of fiber optic cables involves meticulous measurement of the UV detector’s front and the placement of the flow cell. Maintaining an appropriate bend radius is essential to prevent damage to the delicate glass fibers within the cable casings. Despite the rugged appearance of the casings, it’s crucial to remember that the fibers themselves are made of glass, requiring careful handling to ensure their integrity and longevity.

In conclusion, the integration of UV detectors with remote flow cells represents a significant advancement in liquid chromatography technology. The benefits include reduced dead volume, minimized peak broadening, and enhanced versatility in various chromatography applications. As technology continues to evolve, the synergy between UV detectors and remote flow cells is likely to play a crucial role in pushing the boundaries of liquid chromatography, opening doors to new possibilities in fast protein liquid chromatography (FPLC), preparative liquid chromatography, process liquid chromatography, the separation of PET isotopes, flow chemistry applications and beyond.

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High Performance Liquid Chromatography (HPLC) pumps are an essential component of HPLC systems. HPLC pumps are used in a variety of analytical and research applications.

HPLC pumps can be categorized into two types: isocratic and gradient. Isocratic pumps deliver a constant mobile phase composition throughout the analysis, while gradient pumps allow for the composition of the mobile phase to be varied over time, allowing for greater separation of components. Both types of pumps require precise control over the flow rate and pressure of the mobile phase to ensure accurate and reproducible results.

HPLC pumps are typically composed of three major components: the pump heads, drive mechanism piston, and the motor. The pump heads contain the inlet and outlet ports for the mobile phase, the check valves responsible for maintaining the flow direction and the piston is responsible for moving the mobile phase through the pump head. The motor and drive mechanism provides the energy to drive the piston. The pump head and piston are typically made of materials that are resistant to the mobile phase and can withstand high pressures, such as stainless steel and titanium.

One of the key considerations in the selection of an HPLC pump is the flow rate and pressure range required for the analysis. HPLC pumps can deliver flow rates ranging from a few microliters per minute to several liters per minute, depending on the application. The pressure range required for the analysis is typically determined by the type of column and the particle size of the stationary phase. Columns with smaller particle sizes require higher pressures to maintain the flow rate, and therefore require pumps capable of delivering higher pressures.

Another consideration in the selection of an HPLC pump is the type of mobile phase being used. Some mobile phases, such as high-viscosity solvents or viscous solutions, can require more powerful pumps to maintain a constant flow rate and pressure. In addition, the compatibility of the pump materials with the mobile phase should be considered, as certain materials may be incompatible with certain solvents or additives.

Maintenance and care of the HPLC pump are critical for ensuring reliable and reproducible results. Regular maintenance, such as cleaning and the pump head and replacing pistons, seals and check valves can help prevent contamination and ensure smooth operation. In addition, regular calibration and validation of the pump, including flow rate and pressure measurement, can help ensure accurate and precise results. The use of high-quality mobile phases and filters can also help prevent contamination and prolong the life of the pump.

One of the challenges in HPLC pump operation is the potential for system pressure fluctuations, which can lead to inaccurate and unreliable results. Pressure fluctuations can occur due to a variety of factors, including air bubbles in the mobile phase, clogged filters, or leaks in the system.

The HPLC pump is a critical component of the HPLC system responsible for delivering the mobile phase at a constant flow rate and pressure. The selection, maintenance, and care of the pump are critical for ensuring reliable and reproducible results

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Degassing is the process of removing dissolved gases from a liquid. In FPLC, online degassing is achieved by passing the mobile phase through a degasser unit before it enters the column. The degasser unit uses vacuum pressure to remove dissolved gases such as oxygen and carbon dioxide, which can interfere with the protein purification process. This allows for the mobile phase to be free of any dissolved gases, resulting in improved protein yield and purity. Degassing the mobile phase prevents bubble formation. An online degasser is the preferred method.

One of the main benefits of online degassing is that it helps prevent protein denaturation. Denaturation is the process by which a protein loses its native structure and function. This can be caused by several factors, including changes in pH, temperature, and the presence of denaturants such as detergents or salts. However, dissolved gases can also cause denaturation by altering the pH of the mobile phase. When carbon dioxide dissolves in water, it forms carbonic acid, which can lower the pH of the mobile phase. This can cause proteins to become denatured, leading to reduced yield and purity.

In addition to preventing denaturation, online degassing also helps prevent protein aggregation. Aggregation is the process by which proteins form clumps (aggregates). Which can reduce the yield and purity of the protein sample. Dissolved gases can cause aggregation by altering the electrostatic interactions between protein molecules. For example, oxygen can react with sulfhydryl groups on proteins to form disulfide bonds, which can lead to protein aggregation. By removing dissolved gases, online degassing helps prevent these unwanted interactions and ensures a higher yield and purity of the protein sample.

Online degassing is also important for maintaining the accuracy and reproducibility of FPLC experiments. When dissolved gases are present in the mobile phase, they can cause fluctuations in the baseline of the chromatogram, making it difficult to accurately measure the absorbance of the protein sample. This can lead to inaccurate quantification of the protein yield and purity and can even result in errors in downstream applications such as protein structure determination. By removing dissolved gases, online degassing helps ensure that FPLC experiments are more accurate and reproducible.

Another benefit of online degassing is that it helps improve the stability of the protein sample. Dissolved gases can cause oxidative damage to proteins, leading to reduced stability and increased susceptibility to proteolysis. This can be particularly problematic for labile proteins or proteins that are sensitive to oxidative stress. By removing dissolved gases, online degassing helps protect the protein sample from oxidative damage, improving its stability and shelf life.

Online degassing is critically important for single pump gradient FPLC systems. In these systems the gradient is formed before the pump. Degassing before the valve inlets enables buffers to be bubble free. Bubbles in the valve inlet create inaccurate and irreproducible gradients.

Finally, online degassing can help reduce the cost and time associated with FPLC experiments. When dissolved gases are present in the mobile phase, they can cause column fouling, leading to a decrease in the efficiency of the chromatography. This can result in longer run times and increased use of mobile phase, which can be costly and time-consuming. By removing dissolved gases, online degassing helps prevent column fouling and ensures a more efficient and cost-effective FPLC experiment.

In conclusion, online degassing is an essential step in FPLC that helps prevent protein denaturation and aggregation, maintains the accuracy and reproducibility of experiments, improves the stability of the protein sample, and reduces the cost and time associated with FPLC experiments. By removing dissolved gases from the mobile phase, online degassing increases the reliability of the FPLC system.

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There are many types of high-performance liquid chromatography (HPLC) detectors. HPLC UV detectors are used to detect, analyze and identify the components of an eluted mixture after it is run through an HPLC column. The most used HPLC detectors are ultraviolet (UV) detectors. UV-VIS (ultraviolet visible) detection is used with HPLC to detect and identify analytes in a sample. UV detectors use light to analyze the sample. The analyte is identified by measuring the sample’s absorption of light at different wavelengths. UV-VIS detectors work by passing visible and UV light through a sample in a flow cell. The UV detector measures the sample’s absorption of light at different wavelengths as it passes through the cell.  This process identifies the analyte. The properties of the sample of interest are determined by the amount of light absorbed. UV detectors are easy to use, reliable, and have a universal response to chromophoric compounds.  UV detectors are in-line detectors that measure the UV absorbance of the HPLC eluent, they provide a continuous signal used to identify and quantify the amount of chromophoric compound coming out of the HPLC column. For all these reasons, UV detectors are the most common detector used in an HPLC system. UV detectors are also employed in FPLC (Fast Protein Liquid Chromatography), Preparative HPLC, UHPLC (Ultra High Performance Liquid Chromatography), as well as flow reaction chemistry monitoring.

UV detectors employ a deuterium lamp (D2 lamp) as the light source. A D2 lamp detects wavelengths from 190-400nm. If higher wave length detection is needed, a UV-VIS detector is used with an additional tungsten lamp. A xenon lamp can be used to cover the entire wavelength range from 190-800 nm.

The advantages of a Xenon source for a UV/VIS detector

What is the ideal detector design for protein purification, PET isotope purification, preparative chromatography as well as process chromatography? 

The best UV detector design would incorporate the following elements:

• Xenon light source
• smallest possible footprint 
• fiber optic flow cells for remote sensing
• wide variety of flow cells & materials to handle all applications 
• the ability to monitor up to 4 wavelengths simultaneously 

Xenon is the ideal source for UV detectors in all applications where ultimate sensitivity is not required. 

A benefit of a xenon source is that it requires only 20% of the power typically needed for a deuterium light source. A Xenon lamp requires minimal warm up time compared to a deuterium lamp which takes 20-30 minutes to warm up. An additional advantage of using a Xenon light source is that it produces very low heat, so it is warm to the touch vs using a deuterium lamp, which is burning hot to the touch. The heat from a deuterium source can potentially denature proteins. In addition, the Xenon source covers the UV and visible range from 190 nm up to 800 nm as compared to a deuterium (D2) source which performs well from 190 nm to 400 nm. When a deuterium lamp is used as the light source, a tungsten halogen lamp needs to be added in order extend the range into the visible. Another benefit of a Xenon source is that the lamp and the power supply are housed together. It is very simple to replace both the lamp and power supply as one unit (making it plug and play), vs. replacing a D2 lamp, where the lamp and D2 power supply are separate units. Replacing a D2 lamp requires more time, effort and cost when they fail. The life expectancy of the Xenon source is in excess of 2.5 times the life expectancy of the deuterium lamp. Over time, the operating cost of the Xenon lamp is forty percent of the cost of operating a deuterium lamp, not even accounting for the time and effort required to replace the additional deuterium lamps. 

The benefits of a Xenon source for a UV detector are significant. The Xenon source represents the best alternative to a deuterium source in variable and multi-wavelength UV-VIS detectors used for protein purification, PET isotopes, preparative chromatography, process chromatography and also in HPLC where ultimate sensitivity is not required. 

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High Performance Liquid Chromatography (HPLC) is the gold standard in analytical and preparative chemistry. Liquid chromatography systems pump solvents or buffers through the system enabling the compounds of interest to be separated and identified. HPLC is a powerful and very precise technique that has broad applications throughout many fields. HPLC is used to identify and provide quantitative analysis of any drug or compound, detect metabolites, and separate and purify compounds of interest. HPLC is commonly used in food and beverage quality control, clinical, forensic, environmental water purification and for quality control in pharmaceutical drug development.

High Performance liquid chromatography involves pumping buffers or solvents into a fluid stream. It is critically important that bubbles and gases are removed prior to entering the fluid stream. The most efficient and cost-effective method to accomplish this is membrane degassing. ICON Scientific offers a full range of membrane degassers that can handle analytical flow rates up to 1 liter.

Degassing removes dissolved gas from fluids before they outgas, reducing the chance of bubble formation. On-line degassing is an essential technique when doing HPLC, uHPLC, FPLC, ion chromatography and HPLC mass spec.

On-line degassing is valuable because it replaces helium sparging, improves quantitative analysis and eliminates bubble formation which creates problems for HPLC, uHPLC and FPLC systems.

OUTGASSING

“Outgassing” is a term used to describe dissolved gases emitted from a solution. Outgassing occurs in HPLC systems when rough surfaces produce nucleation sites for bubbles to form. Outgassing occurs anytime gases dissolve into a solvent or buffer and cause bubbles to form in pumps and detectors. Bubbles entering the HPLC system will get trapped in the detector flow cell sending spurious signals to the detector. Bubble formation in a flow cell that is caused by outgassing will create false peaks and noisy baselines. Outgassing disrupts the column bed in FPLC ruining the separation process.

      High- and Low-Pressure Mixing

There are two ways to create a gradient in HPLC. There are low-pressure and high-pressure gradients. 

A low-pressure gradient, sometimes called a ternary or quaternary gradient, is when the mixing occurs with a low- pressure proportioning valve. The proportioning valve is placed prior to the liquid entry to the pump. This type of gradient formation is called low pressure because the mixing takes place on the low-pressure side of the pump. Outgassing negatively affects gradient formation because the proportioning valves are time based and cannot distinguish whether liquid or air bubbles are being pumped. Low pressure mixing systems are more prone to bubble formation. 

High pressure mixing is sometimes referred to is a binary gradient system. High pressure mixing occurs when the gradient formation is produced by two pumps controlling the relative rate of pump A verses pump B to produce a constant flow rate. In high pressure mixing the mixing occurs after the pumps and takes place in the mixing chamber. High pressure mixing is less prone to outgassing because in this configuration each solvent has its own pump, and the mixing takes place after the pumps. Bubble formation can likely be the cause of many performance issues with both high- and low-pressure mixing.

https://www.iconsci.com/product-category/degassers/

The Solution

Degassers are the solution to removing dissolved gases from fluids before they outgas. Once bubbles are formed, they affect the precision, accuracy, and performance of your chromatography system. Degassing will prevent the formation of problem causing bubbles. Using a degasser will improve precision and reliability and avoid production disruptions or delays. Our degassers optimize the performance of HPLC, uHPLC and FPLC systems. 

            Benefits of Degassers

• Prevent bubble formation
• Efficiently removes dissolved gas from fluid streams
• Improve baseline stability
• Reduce noise
• Reduce start-up times
• Reduce errors by removing gas
• Ensure accurate and constant results
• Improves and optimizes all applications (analytical, biotech or diagnostic)

DUAL FILM DEGASSERS

ICON Scientific has introduced a line of HPLC degassers utilizing patented dual film degasser chambers. Our degassers have the highest efficiency with the lowest pressure drop in the industry. ICON’s degassers are compatible with most HPLC solvents and are biocompatible for FPLC as well.

Our thin film degasser uses a high-efficiency dual membrane and unique flow channel design. Like ICON’s other products, the iPLANAR degasser has a tiny footprint. The iPLANAR offers better performance and the lowest fluidic resistance in the industry. The iPLANAR on-line degasser modules employ proprietary technology. We incorporate a flat membrane on both sides of the cassette to provide maximum surface area with minimal volume. The efficiency is optimized by precisely controlling its thickness to ensure maximum degassing efficiency. The membrane has a high gas permeability therefore optimizing vacuum degassing and providing continuous, efficient gas removal, regardless of flow rate, ICON’s iPLANAR degasser provides dramatic improvement in performance over tube-based degassers. iPLANAR’s advantages include:

• Lowest pressure drop compared to tube-based degasser due extremely low flow restriction
• Bio inert metal free flow path providing virtually universal HPLC solvent compatibility as well as FPLC buffer systems
• NO internal fittings so NO risk of internal leakage
• Higher surface areas
• Highest efficiency degassing
• Lowest internal volumes
• Superior bubble removal and faster pull-down times
• Analytical to semi-prep
• Dual flow plates sandwiched between two vacuum plates coated with Teflon™ AF thin films
• Option of two different size cartridges depending on desired flow rate.
• OEM configurations available

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ELUENT HEATERS generate a constant temperature for the solvent prior to entering the column.

COLUMN THERMOSTATS are available with different dimensions to enabling them to handle anywhere from one column to many columns, and from analytical to preparative columns.

To enable reproducible chromatography from run to run, it is mandatory that the column and solvent within the column are kept at a constant temperature.  The column temperature must be stable not only for reproducible separations but for constant retention times. Increased column temperature causes a decreased solvent viscosity and therefore a lower backpressure. This enables your HPLC systems to run at higher flow rates resulting in faster analysis times.

ELUENT HEATING devices monitor and control the temperature of the eluents. It is important to have full control and overview of temperature settings and the actual temperature of the eluent.

WHY ARE BOTH COLUMN THERMOSTATS AND ELUENT HEATERS CRITICAL TO ACHIEVE REPRODUCIBLE HPLC RUNS?

It is extremely important that the column temperature is not only constant from run to run but that it remains consistent throughout the column. Separation temperature (column temperature) is instrumental in High Performance Liquid Chromatography (HPLC) since it can affect retention time, selectivity, and peak shape. 

It is critical that the mobile phase (solvent) remains at the same temperature when entering the column, throughout the column and then as it exits the column. An accurate heating temperature must remain consistent from the solvent entering the column body, to the column body itself, and be precisely retained throughout the separation. Heating the mobile phase (solvent) without heating the column creates a radial temperature gradient whereby the center of the column is at the highest temperature and the temperatures gets cooler as it approaches the inside of the column wall. Conversely, heating just the column body creates an opposite radial gradient with the hottest temperature being on the inside of the column wall and the mobile phase is the coolest. If only the mobile phase is heated a longitudinal gradient is formed, with the hottest part being at the entrance to the column and diffusion throughout the column makes the outlet cooler. 

WHAT IS RETENTION TIME?

Retention Time (RT) in High Pressure Liquid Chromatography (HPLC) is the time between sample injection to sample detection, most often with a UV detector.

WHY IS COLUMN TEMPERATURE SO IMPORTANT?

Proper column temperature control is essential for separations with marginal resolution. If the temperature of the column remains constant, retention times remain constant. Consistent retention times enable the chromatography data system to identify and properly quantify any components of interest.

BENEFITS OF COLUMN THERMOSTATS:

• Column thermostats increase temperature above the highest potential ambient temperature. The benefit of this technique is that it assures that the chromatographic run is performed under the same temperature every time. Ambient temperatures deviate from day to day and at different parts of the day, leading to inconsistent retention times causing inaccurate results.
• A significant increase in temperature reduces the viscosity thereby decreasing the back pressure in the column and allowing faster flow rates and decreased run time.
• A less common technique is to perform an isocratic separation using a temperature gradient to separate the compounds.

WHAT IS COLUMN TEMPERATURE?

COLUMN TEMPERATURE is a critical parameter in separations and affects the mobile phase viscosity and the analyte transfer between the mobile and stationary phase.

HOW DOES TEMPERATURE AFFECT ELUTION?

As temperature increases, retention time decreases. At higher temperatures, as retention time decreases, faster elution is possible. At lower temperatures, retention time often increases, especially in reversed phase separations. So, what is ELUTION? ELUTION is the chromatographic process of extracting one material from another by washing it with a solvent.

DOES TEMPERATURE AFFECT SEPARATIONS IN CHROMATOGRAPHY?

Temperature can affect the separation of components in all types of chromatography. This is because as temperature rises, the heat transfers into the solvents which promotes faster and more consistent run times.

ICON Scientific offers a unique preparative combination eluent heater and column jacket. This allows control of the temperature of preparative High Pressure Liquid Chromatography (HPLC) columns. Contact block heaters and convection ovens are available in different sizes and from different manufacturers. The column ovens can be controlled by chromatographic software. Some heaters are available with keypad control. Mobile phase heaters are optional on all units. ICON also offers peltier column thermostats, which allow the ability to cool as well as heat.

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All liquid chromatography including uHPLC, HPLC, FPLC, ion chromatography and LC mass spec benefit from on-line degassing
Other online degassers that are available in the marketplace use a tubular membrane in a cassette. A vacuum pump pulls the gas out of the liquid through the tubing to accomplish continuous gas removal. This has been the de facto standard for many years.  Several years back, standard Teflon tubing was replaced with Teflon AF tubing. This improved both the permeability and the efficiency of the degassing process.

Icon Scientific, Inc. introduces a line of the highest efficiency online hplc degassers in the marketplace. These can be used for uHPLC, HPLC, FPLC, ion chromatography and hplc mass spec. Our degassers use a patented planner degassing chamber. This state-of-the-art technology utilizes an entirely different internal structure while maintaining a very small size.  This arrangement has single flow plate sandwiched between two vacuum plates coated with Teflon AF thin films. This dual film plannar degasser technology is so unique it is patented. Tubular degassing Chambers are intrinsically less efficient. Some additional advantages of our degassing chamber is our Superior bubble removal and faster pull down times. We have the highest efficiency with the lowest pressure drops enabling our system to have the lowest internal volumes in the industry. Our systems can be used with reverse phase and normal phase HPLC solvents as well as a buffer systems used in FPLC, because only PEEK and Teflon AF are used in the flow path.

All other brands of degassers contain internal fittings. Icon Scientifics online dual film planar degasser has no internal fittings or connections in the degasser chamber. Internal fittings have been eliminated on our design to avoid any risk of internal leakage. Dual film planar degasser modules offer the highest efficiency degassing with the lowest pressure drop across the system. This enables our systems to have the lowest internal volumes available in the marketplace. Icon’s dual film planar degasser technology is available in two different size cartridges depending on the flow rate desired.

Icon Scientific offers Superior technology and performance at a similar price point compared to ordinary degasses.

 Choose Iplanar, take the plane vs. the tube.

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 When creating a vaccine, you need to attack the protein spikes on the SARS-CoV-2 virus. Proteins on the SARS-CoV-2 spikes have become the targets for creating vaccines and anti-viral monoclonal antibodies. Before analyzing the proteins of interest, they must be purified in solution. FPLC has been the gold standard for protein purifications for many years. Many prominent researchers create a multi-step purification protocol using FPLC resins and FPLC systems. ICON Scientific offers complete solutions, including a wide range of FPLC components that can be expertly configured to suit your research application. 

As the battle against SARS-CoV-2 virus continues to grow and enter new stages, researchers are relying on liquid chromatography techniques to provide accurate, efficient methods for analyzing potential vaccines and therapeutics.

Liquid chromatography (LC) is a critically important tool in therapeutic research, forming the basis of clinical trials based on RNA purification for the novel therapeutic vaccines. HPLC has been instrumental in pioneering research into the composition, method of action and structure of SARS-CoV-2. High Performance Liquid Chromatography (HPLC) is critical for quality control and quality assurance for the bioactive ingredients used in vaccines as well as therapeutics and all aspects of pharmaceutical development.

Liquid chromatography is a vital technique in many industries including pharmaceutical, production, MRNA production and development, cannabis/hemp analysis, food and beverage industries, manufacturing, quality control and more.

Vaccines need to be purified and filtered at different stages. One of the most important purification steps involves separation techniques using chromatography columns. In liquid chromatography the dissolved liquid (mobile phase) carries the analyte of interest through the column which is packed with a fixed material (stationary phase). Different chemical properties of the molecules within the analyte determine the retention time for the components of interest.

Liquid chromatography (LC) is an essential technique used for the purification of biomolecules including RNA. SARS-CoV-2 is an RNA-based virus. RNA purification uses liquid chromatography (LC) because it is performed at both higher temperature and higher pressure than normal LC. Higher pressure and higher temperature stimulate the denaturation process to remove protein complexes. The combination of high pressure and temperature and a binary solvent gradient prevent RNA degradation, while providing an effective RNA isolation. Liquid chromatography instruments can isolate the targeted messenger RNA, the mRNA and therefore analyze the vaccines efficacy.

Preparative HPLC pumps are used in the manufacturing of vaccines. Liquid nanoparticle (LNP) production equipment is used for the development of mRNA-based vaccines, lipid nanoparticles (LNPs) have proven to be a suitable delivery form. Lipid nanoparticles protect and encapsulating the active ingredient allowing it to be transported to the target cells in the human body. SARS-CoV-2 vaccines are the first large-scale production vaccines using lipid nanoparticles and mRNA in vaccine history. Icon Scientific supplies custom solutions for both small and large-scale production of vaccine lipid nanoparticles.

SARS-CoV-2 researchers are constantly fighting a battle as they screen potential antiviral treatments and continue to perfect vaccine development. 

During these unprecedented times ICON Scientific can help you configure and customize liquid chromatography (HPLC) systems to assist your research into the structure of SARS-CoV-2 and its method of action.

Any Questions? Call us at 301-330-4266

Or send us an email at Icohen@iconsci.com

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LNP’s, nanoparticles with an outer shell of lipid molecules have the ability to encapsulate and transport complex active ingredients (such as mRNA, RNA, siRNA, or DNA-based APIs)) to target cells in the human body. This technology can be precise tuned and targeted. This has led to an expanding demand for their production. This is the methodology that is currently being used to fight against the Coronavirus. ICON Scientific is playing an active role in supporting pharmaceuticals in their mRNA vaccine production. This technology has potential for other vaccines, treatments, medications, and gene therapy. mRNA will continue to grow as a drug delivery system. 

The critical ingredient in mRNA vaccines is the mRNA, the short-lived strands of genetic material that message our cells to begin making SARS-CoV-2 proteins. This signals our immune systems to begin developing antibodies.

Over the past decade, SLNs have been studied as a drug delivery system for controlling drug release. These colloidal carrier systems have the advantages of traditional systems without the major disadvantages. SLNs can control drug release, increase drug stability and targeting, and are easily scaled-up.  Therefore, the development of lipid-based nanoparticles (liposomes in particular), have proven to be highly effective in cancer treatments. They improve the selectivity of chemotherapeutics agents, offer a controlled and prolonged release of agents, and they increase the solubility of hydrophobic drugs. Patients benefit from nanoparticle formulations because they have higher efficacy and reduced side effects.  LNP’s are new from a production point of view, yet the technology has been clinically proven for almost 30 years. LNP-based formulations can be rapidly developed and scaled-up into a finished product. This has been a tremendous advantage in the development of mRNA vaccines for COVID-19. LNP formulations have become recognized as the gold standard for parenteral products such as antibiotics, anticancer agents, and drug combinations. LNP-based drugs have been approved since 1995 with the approval of the lipid based anti-cancer drug, Doxil (doxorbubicin). In 2018, the FDA approved Onpattro, the first nanoparticle drug to be administered intravenously for the treatment of peripheral nerve disease. 

It is promising that mRNA technology could revolutionize vaccination formulization, protein replacement therapies and produce more efficient treatment for genetic diseases. 

Icon Scientific’s HPLC pumps are used in the mass production of liquid-based nanoparticles (LNPs) for the drug delivery of mRNA-based vaccines. Precise dosing is necessary to ensure optimal encapsulation. ICON Scientific’s portfolio of HPLC dosing pumps are optimized to meet the needs of the pharmaceutical industry for mRNA vaccine production. Our pumps precisely deliver fluids at a wide range of flow rates and pressures. They have extremely stable liquid flow. There are three choices of pump material: stainless steel, titanium, or PEEK. When our pumps are used in combination with a mass flow controller, exceptional flow accuracy and precision is achieved.

In addition to dosing pump technology, ICON Scientific offers many other chromatography solutions for the pharmaceutical industry.

Check out more information HERE: https://www.iconsci.com/product-category/liquid-nanoparticles-lnp/

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 What does HPLC stand for and why is it the only method used by professionals?

HPLC stands for high pressure liquid chromatography. HPLC separates liquid molecules from one another in the liquid phase under high pressures. The Cannabis/hemp sample must be extracted into the liquid phase prior to injecting the sample into the HPLC system. High pressure is needed because the HPLC column (which performs the separation) has a very small particle size (less Than 3 microns) and a very large surface area which provides the resolution necessary to separate the cannabinoids. High pressure liquid chromatography can operate under pressures of up to 9000 psi or higher with a pulseless flow. HPLC is the only method capable of separating, identifying, and quantifying the cannabinoids in a liquid sample mixture.

Other Methods Fall Short

There are low pressure liquid chromatography systems that use a peristaltic pump and can only handle pressure less than 100 psi. Peristaltic pumps by design create pulses in order to deliver the liquid stream. Systems employing these types of pumps are not capable of using HPLC columns and instead use a solid phase extraction cartridge with large particles that vary in size. These systems are incapable of separating cannabinoids and therefore it is impossible to accurately quantify them.

A proper HPLC system for cannabis and/or hemp should incorporate a column with a smaller than 3 micron particle size that are monodispersed. Monodisperse means that all particle sizes are exactly the same when examined under a microscope.

 It is also critically important that the temperature of the column is heated above ambient temperature and is accurately controlled. This ensures that the cannabinoid peaks / elute off the column at the same time and in the same order every time. Without this precise control you will get erratic results!

The UV detector must operate with a narrow bandwidth at 228 nm.  This can be achieved by a fixed wavelength detector, a variable wavelength detector or a diode array detector. 228 nm wavelength provides the best sensitivity for cannabinoids. UV/LED detectors while being small and stable are not suitable for this application. UV / LED detectors that are not filtered given a broad spectrum of light which is not appropriate for any HPLC system. 

Another feature to look for is either an automated injection valve or for high throughput systems, a cooled auto sampler. Manual injectors can work but are not as reproducible as the other two options.

 An HPLC cannabis and hemp analyzer should include a solvent degasser. This is important to eliminate potential air from entering the HPLC. If air does enter the HPLC, it will cause bubbles that may lead to erroneous results!

 There are two ways to do an HPLC separation. For method development, a gradient system works the best. In a gradient system you use two solvents, increasing the concentration of one will decrease the concentration of the other to enable your cannabinoids to come off the column one by one. Once this method is established, an expert can convert the gradient method to an isocratic method. By doing this, you were using only one mobile phase. Isocratic separations are preferred for a routine, repetitive HPLC, such as cannabinoid and hemp potency analysis. This methodology eliminates the possibility of an inaccurate gradient while providing a more robust method.

 How to choose instrument control and data acquisition

 Traditionally, laboratory HPLC’s are controlled and data is processed by a sophisticated chromatography data station. These work very well for laboratory HPLC where the operator is a chemist or chromatographer and understands how to program the software and evaluate the data after the chromatographic run. Icon Scientific has introduced a simplistic alternative for the non-technical user. The ANSER is a portable, true HPLC cannabis and hemp potency analyzer. It uses a touchscreen control with pre-programmed entries. This method enables an inexperienced operator to control and obtain cannabinoid potencies without the hassle of programming and analyzing data.

How to determine potency

 In order to determine the potency of cannabinoids several steps need to be taken.  First, the sample must be extracted from the plant material and be put into a liquid form. While there are many variations of this operation, the most important part is that it is accurate and reproducible. Before injecting your cannabinoid samples, you must inject reference samples from a reliable source in order to calibrate your HPLC. It is necessary to get samples of the cannabinoid that you have interest in analyzing. Once you have calibrated the HPLC with your reference samples, it is time to run your unknown samples. Your unknown samples are compared and analyzed to the reference standards and the software / firmware will provide your potency analysis. The ANSER is easy to use, accurate, reliable and reproducible.

 There are four steps necessary for cannabis and hemp potency analysis

1.  Extract sample into liquid form
2.  Run reference standards for the cannabinoid of Interest
3.  Run unknown samples
4.  Software / firmware will provide the cannabinoid potency analysis

 There are many portable potency analyzers claiming to be accurate, sensitive, and reproducible. Portable devices that use spectroscopy without HPLC fall short of being able to provide laboratory quality results that are accurate, sensitive and reproducible.

 That is why HPLC is the gold standard for cannabis and hemp potency analysis. It is clear that the ANSER is your best option for a cannabis and hemp potency analyzer.

 There are many options available for laboratory-based HPLC, Icon Scientific offers a wide array of choices to configure a customized laboratory system. Icon Scientific has recently introduced the first miniaturized, portable, rugged, and true laboratory HPLC cannabis and hemp potency analyzer.

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HPLC pumps deliver a consistent flow and constant pressure of liquid for a chemical reaction. HPLC pumps are used for process applications and/or the production of a product.

ICON Scientific’s vast line of pumps all offer precision, reproducibility and ruggedness. Our high pressure dosing pumps are highly accurate and reliable two-piston pumps for pharmaceutical and chemical applications as well as research and method development. Pumps are used for a wide range of tasks in chemical, food and pharmaceutical industries. Pump heads are available in various materials, stainless steel, titanium, and PEEK, Hastelloy C or ceramic. Maximum flow rates for the pump heads range from 100 ml/min, 250 ml/min, 500 ml/min to 1000 ml/min. Pumps can be controlled either by keypad, PLC control or software control. 

All preparative dosing pumps are a two piston reciprocating design which assures maximum reproducibility of flow, lack of pulsation and extreme ruggedness.

Jet mixing technology has outstanding results for small and large scale pharmaceutical production of vaccine lipid nanoparticles. Small desktop devices are available, as well as complete LNP skids for pharmaceutical researchers.

IJM (impingement jets mixing) skids are available for high flow production of nanoparticles (LNP). These systems can be configured with up to 8 parallel mixing units. Each unit consists of two pumps delivering lipid and API streams, one jet mixer and two flowmeters. All skids are constructed of a stainless steel frame, and built on casters to be suitable for CIP cleaning procedures in pharmaceutical production. We offer customizable solutions to meet our customers specifications. HPLC pumps have now become critically important in the fight against COVID-19. HPLC pumps are being used to produce liquid nanoparticles (LNP) for the production of the mRNA vaccines. ICON Scientific offers a line of HPLC pumps that are extremely suitable for liquid nanoparticle production.

Lipid Nanoparticle

Lipid nanoparticles (LNP) are the delivery form of choice for mRNA based vaccine development and production. LNPs encapsulate and protect the sensitive active ingredient allowing it to enter the target cells in the human body. COVID-19 vaccines are the first large-scale use of lipid nanoparticles (LNP) and mRNA in vaccine history. LNPs can form a stable basis for the administration of RNA, mRNA, siRNA, or DNA based APIs. These active ingredients optimize delivery to the target cells. This is a promising development in medicine.

Lipid nanoparticle (LNP) systems are used to encapsulate mRNA with lipids. The mRNA needs to be protected with lipid production to prevent degradation and to increase integration in the bodies cells.

ICON Scientific’s HPLC experience and sophisticated pumping technology is critically important in enabling us to provide superior LNP solutions.

Very precise pumping technology is extremely critical for high-quality lipid nanoparticle (LNP)  production systems. Our systems offer fast, precise pumping technology, reliable switching of liquids, constant control of parameters using software and a flow meter. The software executes control function when critical changes in the flow rate are recorded. We have an elaborate mixing technology.

 Successful COVID-19 vaccine technology has created significant interest in using mRNA vaccines in place of traditional vaccines. Traditional subunit vaccines have been used successfully but they do not induce cellular immunity. Cellular immunity is needed to eradicate the intracellular pathogen reservoir in viral infections. Live attenuated vaccines are the most potent in activating the immune system, both cellular and humoral immunity. The problem is these vaccines cause considerable safety concerns. In immunosuppressed patients there’s always a concern about administering a live vaccine. Attenuated pathogens, on rare occasions, could revert to the pathogenic form and cause disease.

 mRNA vaccines seem to combine the advantages of live attenuated vaccines and traditional subunit vaccines without the associated risks of live attenuated or DNA vaccines.

 DNA and mRNA vaccines are similar but different. The main difference is the target location for delivery of the oligonucleotides. Oligonucleotides (oligos)  are short single strands of  synthetic DNA or RNA. DNA therapeutics need to reach the nucleus, while mRNA therapeutics target the cytosol. mRNA therapeutics are easier to deliver because they do not need to cross a nuclear membrane. mRNA does not integrate itself or alter the genome. Cytosolic mRNA has no interaction with the genome. Essentially, mRNA represents minimal genetic information, and is only expressed until the mRNA has been degraded. mRNA has the ability to encode multiple proteins having very difficult chemical and physical properties, without affecting its physiochemical properties.

 In conclusion, mRNA synthesis and purification are not only fast, but easy to manufacture and are low in cost  in comparison to other vaccine technologies. Today, lipid nanoparticles are the chosen vectors for in vivo RNA delivery.

 Our wide portfolio of dosing pumps are optimized to meet your needs. The resulting integration of our pumps and mass flow controllers ensures optimal conditions for lipid nanoparticle formation. We have quickly adapted to meet the needs of our pharmaceutical partners.

 In addition to offering hardware and software needed to implement lipid nanoparticles (LNP) scale-up and production, ICON Scientific has over 40 years experience in HPLC pumping systems. We are subject matter experts. Use our experience to your advantage!

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Chromatography is a laboratory technique used to separate compounds from a mixture in order to purify or identify them. The science of chromatography originated over 100 years ago. Chromatography is a rapid and efficient separation technique. Chromatography was first used primarily for the separation of plant pigments such as chlorophyll and carotenoids. Mikhail Tsvat was the first scientist to use chromatography. He separated plant pigments into bands of color by injecting plant extracts mixed with petroleum into a glass column that had calcium carbonate in it. As the liquid passes through the column, bands of color were separated. This enabled them to identify the compounds of the plant. Through the beginning of the 20th century, “color writing” or chromatography was used to separate plant pigments such as chlorophyll (green) and carotenoid (orange/yellow). By 1930, scientists had figured out they could use these techniques for other separation processes and chemical analysis, especially in biochemistry. 

Liquid chromatography 

In 1952, Martin and synge won the Nobel Prize for their invention of partition chromatography. This was the basis for liquid chromatography. High pressure liquid chromatography (HPLC) has two phases, a stationary and a mobile phase. The packing material, typically called a resin is referred to as a stationary phase because the resin remains fixed in the column. The mobile phase, as it sounds, moves. The reservoir holds the solvent which is referred to as the mobile phase. The mobile phase is pumped through the column with the HPLC pump. The sample is separated based on the resonance time inside the column with the output monitored by a detector, usually a UV detector which creates a chromatogram. High pressure liquid chromatography is the most popular testing method. HPLC is the best method for cannabis and hemp potency testing because it provides the most accurate, consistent analysis.

What is required to perfect a chromatographic separation?

In order to perfect a chromatographic separation, the peaks which are components of the mixture need to be completely separated or resolved from closely eluting compounds. 

An ideal high pressure liquid chromatography system must consist of the following components. Since the sample goes in the injector, we will start there. The injector should be automated with a fixed loop. This will provide consistent injection volume. The mobile phase should be degassed with an online degasser assuring that the pump is only pumping liquid without air bubbles. 

The HPLC pump should be a two piston high pressure HPLC pump capable delivering pulseless flow at the flow rate needed for the application. Typical HPLC pumps can handle up to about 600 bar of back pressure. This is important because the HPLC column should have a uniform size particle (solid phase) which should be less than three microns in size. This ensures the highest resolution, best sensitivity, and reproducibility of chromatograms. 

Another critical factor is making sure you have a column heater. A column heater allows the column to run at the same temperature every time. This assures run to run reproducibility and consistent chromatograms. If the column temperature varies, retention times change possibly leading to inaccurate identification of peaks and their quantification. 

The UV detector for cannabis and hemp should be focused at 228 nm. Unfiltered UV LED detectors are not able to give quantitative results because they are looking at broad spectrum UV data, all of which is above the maximum absorption of cannabinoids, which is at 228 nm.

In conclusion… 

Icon Scientific’s ANSER is an analytical grade HPLC. HPLC is the gold standard for cannabis and hemp potency testing and is the technique used in certified contract laboratories. 

The ANSER is extremely portable and ruggedized. The ANSER is a purpose built, simplistic, user-friendly potency testing solution for cannabis and hemp. The ANSER allows you to determine harvest time, potency and safety. This is critically important whether you are a researcher, farmer, processor, government agency, product developer, or work in law enforcement. The ANSER is a true miniaturized HPLC. It is a turn-key design intended to be used by non-chemist and non-chromatographers.  

Call or visit our website for more specs and details. Icon Scientific offers a full complement of HPLC equipment as well as CPC, CCC, flash chromatography, a non-destructive cannabis extraction system, and a process scale purification system.

CALL US: 301-330-4266

VISIT our website: www.iconsci.com

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Icon Scientific has designed and built analytical grade HPLC components. These HPLC components have been miniaturized, simplified, and ruggedized. Icon Scientific has also created a simplified graphic user interface (GUI) so that a non-chemist, non-chromatography can easily operate the system with minimal training. All the components are housed in a military grade case. The case is built for hard use and is waterproof. It is made of an ultra-high strength polypropylene copolymer resin, enabling it to be water and dust tight.

What is in the box?

The HPLC pump is a dual piston pump that delivers pulseless flow at up to 9000 psi of back pressure. This is important because the HPLC column has extremely small particles that are uniform and less than three microns in size. The column housing is a stainless steel tube approximately 6” long. A pump of this caliber is necessary to push the solvent through the column at a high back pressure in order for all the cannabinoids to be completely separated. Incorporated in the system is a focused UV detector aimed at 228nm. Analytical research shows that 228nm is the best wavelength to measure cannabinoids. It is also important for the solvent to be degassed before it is pumped. Degassing the solvent assures that you have the most accurate flow rate possible. HPLC separations vary depending on temperature. We have incorporated a column heater to assure run-to-run reproducibility over time. Sample injection is another critical area that needs to be addressed. We assure accuracy based on our motorized valve injector. 

Why is HPLC important?

HPLC (High Pressure Liquid Chromatography) is the gold standard for cannabis and hemp analysis. It is the only method used in certified contract laboratories. HPLC accurately separates, measures, and differentiates targeted cannabinoids. The ANSER achieves the sensitivity, reproducibility, and reliability of a bench size analytical HPLC. Additional benefits are the ANSER’s size, it being fully portable and easy to use. 

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                Cannabis and hemp testing has become critically important and an integral part of being able to determine a products potency and safety.

                Cannabis potency testing enables the user to quantitate the major cannabinoids. At this point, the ANSER can identify and quantitate 18 cannabinoids in less than 7 minutes. The 18 cannabinoids include: CBDVA, CBDV, CBDA, CBGA, CBG, CBD, THCV, THCVA, CBNA, CBN, EXO-THC, D9-THC, D8-THC, THCA-A, CBCA, CBL, CBC, CBLA.

                Potency analysis identifies compounds and measures their level of strength (potency). Potency testing is typically performed using HPLC. HPLC stands for High Performance Liquid Chromatography. HPLC is an analytical technique used to separate, identify, and then quantify components in a mixture. HPLC is an essential technique in labs worldwide. HPLC is the gold standard used by contract potency testing laboratories because it is the most reliable and accurate method of measuring cannabinoids. Liquid Chromatography used with a UV detector enables the user to identify the potency of the cannabinoids. It is critical to have fast, precise potency testing to measure the strength of a product or time a harvest. HPLC testing is the only accurate way to measure the potency of cannabinoids in cannabis and hemp.

                The ANSER is the perfect solution for laboratories, manufacturers, cultivators, extractors, growers, product manufacturers, regulators, law enforcement agencies, hemp compliance, and quality control labs to monitor cannabis and hemp potency. The ANSER will enable you to increase quality, consistency, and consumer safety. 

                Real HPLC done right is not complicated. The ANSER is a turn-key solution. It has a simple touchscreen control. There is no complicated chromatography to learn. It is intended for use by a non-chemist and non-chromatographer. Everything is built-in. The ANSER is a self-contained system in a ruggedized, military grade PORTABLE case. 

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High Performance Liquid Chromatography (HPLC) is the industry standard or gold standard for cannabinoid potency testing. It is the most accurate and reliable method to measure cannabis components. HPLC is the acceptable standard used by certified contract labs.

ICON Scientific’s ANSER is your solution for cannabis potency and purity testing. The ANSER is the only true HPLC based analyzer. We are committed to supplying growers, processors, dispensaries, laboratories, and product developers with HPLC solutions for accurate and reproducible cannabis and hemp analysis.

The ANSER is a portable, easy-to-use potency and purity analyzer for the cannabis and hemp industry. No chemistry or chromatography knowledge is required to operate this analyzer. The ANSER is always precise, accurate and reproducible. The fully portable ANSER allows you to test the potency of samples in the field or in the lab.

The ANSER is not only portable, but a miniaturized, turn-key HPLC solution. It has all the performance of a benchtop HPLC without the size. The ANSER includes a column, mobile phase, certified standards and methods.

The ANSER analyzer provides an immediate answer.

ICON SCIENTIFIC, Inc. Introduces…

The ANSER™ Cannabis & Hemp Analyzer

The ANSER can be operated anywhere you need to with our battery pack option.

Intuitive software control allows the non-chromatographer to operate and easily evaluate his data.

We supply methods:

1. For maximum THC concentrations to enable growers to know exactly when to harvest their plants.
2. For purity of CBD. To ensure there is no THC in your CBD product.
3. For purity of THC.
4. Testing of incoming THC or CBD before final product is produced.
5. Testing of finished product for either CBD or THC.

HPLC stands for high performance liquid chromatography. HPLC is used for the identification and/or quantification when determining the potency of cannabinoid components. Potency analysis identifies compounds and measures their “potency”, level of strength. Cannabis is made up of 500 or more chemical components. Those most commonly tested for are:

• THC (Tetrahydrocannabinol)
• THCV (Tetrahydrocannabivarin)
• CBD (Cannabidiol)
• CBDV (Cannabidivarin)
• CBC (Cannabichromene)
• CBN (Cannabinol)
• CBG (Cannabigerol)
• CBGV (Cannabigerivarin).

The ANSER is the ideal potency analyzer for many applications. The ANSER is the perfect solution for growers, manufacturers, laboratories, regulators and discrepancies to monitor cannabis and hemp potency. The ANSER enables the end user to accurately time harvests, determine the proper profile of the strain and accurately label it. When cannabinoid components are used for medical treatment it is crucial to know the exact components, potency and purity to ensure accurate labeling. Consumers are frequently not getting what they think they purchased.

• HPLC Stands for High Performance Liquid Chromatography
• HPLC is used to identify and / or quantify when determining the potency of cannabinoid components.
• Potency analysis identifies compounds and measures their “potency” or level of strength

HPLC can test plants in the field, edibles and liquids. This is because samples can be tested at lower temperatures than with GC. Using GC heat catalyzed reactions occur. Therefore, HPLC is the “gold standard” for potency testing of cannabis. ICON’s field portable ANSER HPLC analyzer is a simple way for cannabis and hemp professionals to test at every level.

• Turn-key HPLC Solution
• FAST- a Cannabinoid a minute
• Same accuracy and reproducibly as a much larger laboratory HPLC
• HPLC is the “gold-standard” for cannabis potency testing
• Non-chromatographer can operate
• Portable
• Enables you to increase quality, consistency, and consumer safety
• No compromise in quality or performance

…ICON Scientific has the ANSER!

CALL US: 301-330-4266

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ICON Scientific has been an HPLC distributer since 1993. Over 40 years of liquid chromatography experience enables us to be expert in product application, configuration, and customer application. With recent changes in legislation, we saw a unique opportunity to enter the cannabis and hemp space. 

Our vast portfolio of products includes:

Extraction Systems:                                                                                                                                                                                              

• unique technology – natural extraction solution                                                                                                                                                                                                                  
• low pressure extraction technology based on TFE (tetrafluoroethane)                                                                                                                                                 
• TFE’s are very inert, no residue – lower cost – less degradation -yields significantly better extraction and much lower    critical pressures & temperatures

Flash Chromatography:                                                                                                                                                                       

• has the lowest capex                                                                                                                                                                                              
• purify kilograms of material                                                                                                                                                                                             
• handle preparative HPLC columns in addition to flash columns    

Cannabis Purification Systems:                                                                                                                                                

• Preparative HPLC System, capabilities up to 4 liters/min. are available                                                                                                                                
• capable of processing up to 20 kilos of material per day                                                                                                                                                                          
• multiple systems can be set up in tandem                                                                                                                                                                                 
• economical scale-up                                                               

CPC- Centrifugal Partition Chromatography:                                                                                                                                    

• for purification                                                                                                                                                                                        
• stationary and mobile phases are liquid                                                                                                                                                       
• higher purity than traditional HPLC                                                                                                                                                                       
• costs more to purchase but very low operating costs

We look forward to the opportunity to discuss your needs and application. 

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What is HPLC? What do the letters HPLC stand for?

HPLC stands for either High Pressure or High Performance Liquid Chromatography. Chromatography is a separation science. 

How do other techniques fall short?

GC or gas chromatography falls short because this technique requires high temperatures that change the chemical structures of some of the cannabinoids. 

How about standard or low pressure liquid chromatography?

Low Pressure Liquid Chromatography does not have the sensitivity or the resolving power necessary for hemp and cannabis potency testing. There are several reasons why Low Pressure Liquid Chromatography is not an effective technique. HPLC columns are typically about 6” long or longer and they are made out of stainless steel. The separation takes place in the column. In order to push the cannabinoids and solvent through the column, you need an HPLC pump capable of supplying 6000 – 9000 psi of pressure without pulsation. Low Pressure Liquid Chromatography (LPLC) systems usually use peristaltic pumps that by design create pulsation that at best can supply 80 – 100 psi of pressure. The systems are limited to using short fat plastic columns that are only capable of very crude separations with limited analytical quantification. There are many more reasons that LPLC systems are not suitable for this application.

Why don’t spectroscopic methods work for this application?

Spectroscopic methods include all methods that use light waves from infrared to ultraviolet. These methods work best to identify a single chemical. A spectra of a known chemical is stored in the memory. When you scan an unknown chemical, it is compared against your known spectra and when a match is found, your unknown chemical is identified. In a complex matrix such as cannabis and hemp, these methods do not work well. 

If point, click and analyze sounds too good to be true it is.

In conclusion, HPLC is the only method that has the sensitivity, accuracy and resolution that is required for proper hemp and cannabis analysis. 

CALL US for more information: 301-330-4266

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