Continuous manufacturing in Chromatography
High productivity with lowered OPEX by Continuous CPC
Continuous manufacturing and processing has been practised in many industries for decades and has become the primary mode of production. It is widely regarded as the most cost-efficient engineering approach for manufacturing particularly excelling in large-scale production of a single product.
Continuous processing has also been adopted in chromatography, improving productivity and enhancing its industrial applicability while reducing buffer, solvent, and raw material consumption within a smaller footprint. Continuous chromatography can be utilized in various fields, including the chemical industry, pharmaceuticals, biomanufacturing, biotechnology, upstream and downstream processes, and flow chemistry.
In a classic batch process, a quantity of the desired product is manufactured within a single production run - meaning a single sample injection in the case of chromatography - characterized by a well-defined start and endpoint. Once the batch is completed, a new production cycle or run can begin.
On the other hand, continuous chromatography enables higher throughput, improved solvent utilization, and reduced waste generation, making it a more sustainable and cost-effective solution. During continuous processes, the sample injection remains uninterrupted throughout the chromatographic separation, and the process operates without distinct start or endpoint markers. The production of the product in ongoing, resulting in a continuous inflow of raw materials and a constant outflow of the product.
Continuous processing improves efficiency by increasing throughput, while reducing solvent consumption, raw material requirements, and downtime, leading to lower operational expenditures (OPEX) compared to batch processing.
Continuous processing enhances production yield and allows the same equipment to be used for both small- and large-scale operations, minimizing the need for scale-up studies. It is also more energy- and labor-efficient, requires a smaller plant footprint, and is a more environmentally friendly alternative to traditional batch processing.
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Continuous Chromatography Systems:
How the Continuous CPC Works
How CPC, and MDM-CPC work
Centrifugal Partition Chromatography (CPC) is a liquid-liquid preparative chromatographic technique. The separation occurs between two immiscible liquid phases, which are the lower and upper phases of a two-phase solvent system. A strong centrifugal force is applied to immobilize the appropriate phase to function as the stationary phase in the series of cells contained in the rotor serving as multi column chromatography. The other phase in the two-phase CPC system serves as the mobile phase.
CPC operating in batch mode, separates compounds using two different elution methods: ascending (normal-phase) and descending (reversed-phase) mode, all within a single process. This is possible because both the stationary and mobile phases are liquids, unlike traditional chromatography methods that use solid stationary phases.
In descending mode, the upper phase serves as the stationary phase, while the lower phase functions as the mobile phase, which is pumped through the column. In ascending mode, the roles reverse: the upper phase becomes the mobile phase, and the lower phase becomes the stationary phase.
An alternating approach, known as dual-mode CPC (MDM-CPC), maximizes separation efficiency by leveraging both elution modes sequentially. This switching mechanism, along with changes in flow direction and elution order, allows for the effective separation of compounds that have limited selectivity in standard chromatography.
Multiple Dual-Mode CPC (MDM-CPC) in Continuous CPC
Increasing throughput and productivity using two CPCs
In a new continuous chromatography system, two CPC devices are interconnected, and the feed mixture is introduced to the columns in a two-column setup.
The sample injection can be positioned at the front of the rotors, effectively increasing the column length and improving separation efficiency by raising the theoretical plate number (N). Alternatively, injection can be placed between the rotors, enabling a truly continuous (uninterrupted) sample injection mode, making a fully continuous process feasible.
Continuous CPC implements the unique MDM-CPC approach, which involves a sequence of steps where injection remains continuous. In addition, with four built-in pumps, Continuous CPC allows for the alternating use of both CPC modes in a continuous manner (ascending and descending modes). The flow direction and the roles of the upper and lower phases switch multiple times, with each phase acting as the mobile phase at different points, optimizing separation efficiency without interruption by alternating between ascending and descending modes. This design enables the process to be extended into a repetitive cycle, ensuring continuous operation and sustained production.
At the end of the continuous approach process, less-retained and highly-retained components are collected at opposite ends of the column. The key to this approach is inverting the elution mode before any collected product becomes impure, so it maintains the maximal purity, while it minimizes sample loss, and increases the productivity.
Benefits of CPC in continuous process chromatography
Effortless Scale-Up for Higher Production Volumes
Without batch-related constraints, Continuous CPC enables a disruption-free process, simplifying scale-up efforts even in industrial environments with high-load downstream processes.
Exceptionally High Purity
Centrifugal Partition Chromatography (CPC) and MDM-CPC delivers high-purity compounds with no overlapping peaks, ensuring precise separation, isolation, and remediation, including the removal of contaminants and byproducts.
Lower CAPEX & OPEX
✔ No need for silica gel, cartridges, or frequent restarts
✔ Reduced solvent consumption with effective recycling
✔ 3x higher productivity than batch processing
✔ Lower costs compared to MCSGP and SMB
✔ Resin-free purification for industrial-scale applications
Seamless, Nonstop Production
Continuous operation eliminates frequent start-ups and shutdowns, providing consistent sample loading and automated process control, reducing human error and ensuring a steady, uninterrupted workflow.
Smaller Footprint, Higher Efficiency
Compared to traditional chromatography systems, Continuous CPC requires less space while offering higher throughput, allowing greater production capacity without expanding facility requirements.
Advanced Purification for Complex Compounds
With its liquid-liquid nature, CPC enables efficient purification of challenging compounds by allowing customized biphasic solvent systems tailored to the target compound’s unique solubility and chemical properties.
Applications in Continuous bio-manufacturing
In the field of continuous bio-manufacturing, Centrifugal Partition Chromatography (CPC) is a versatile and highly efficient separation method. Its ability to operate continuously enhances efficiency, scalability, and cost-effectiveness, supporting high-throughput bioproduction processes while maintaining product purity and consistency. It provides high-resolution separation, exceptional productivity, and superior purity for binary mixtures while also offering a unique solution for the separation of chiral compounds, lipid nanoparticles, oligonucleotides, and proteins. Continuous CPC is particularly well-suited due to its versatile liquid-liquid nature, making it an excellent choice for flow chemistry applications.
Oligonucleotide isolation
For industrial-scale production of oligonucleotides crucial in vaccine development and pharmacology, CPC can do it in a single purification step.
Purification of isomers
CPC can purify chiral compounds, epimers and diastereomeres (e.g. remdesivir). Purification can easily be scaled up to industrial-scale compared tot raditional chromatographic solutions.
Lipid nanoparticle purification
Lipid nanoparticles (LNPs) are crucial components in mRNA vaccine formulation in pharmaceutical drug delivery research, consisting of key lipid components. CPC can not only ensures the stability of the target lipid but also outperforms traditional chromatography techniques in terms of purity and yield.
Alkaloid remediation
Using RotaChrom’s CPC method and solvent recycling system, you can achieve an increase of 4-5x orders of magnitude in throughput during alkaloid-remediation.
Peptides and proteins
CPC technology has the transformative potential in revolutionizing purification processes of oligopeptides and large proteins in pharmaceutical industries.
Flow chemistry
Flow chemistry, also referred to as continuous flow, involves conducting chemical reactions within a continuous flow stream. CPC can be integrated into a downstream purification process needed for a flow chemistry operation, and the system can benefit from the advantages of both constituents.
Continuous CPC: Triple Productivity in Industrial-Scale Chromatography With Lowered OPEX
Chromatogram of the separation of the components.
Comparison in the total solvent consumption and the productivity between batch and multiple-dual modes.
Case Study: Efficient Removal of Pyrrolizidine Alkaloids from Red Clover Extract
The goal of this separation was to reduce pyrrolizidine alkaloid (PA) content below a critical threshold while preserving the valuable isoflavone content in red clover extract. Using Continuous CPC, we successfully lowered PA levels below the specified limit—without compromising the extract’s beneficial compounds.
Red Clover in Medicine
Red Clover (Trifolium pratense) is widely used in traditional medicine for treating osteoporosis, heart disease, arthritis, eczema, asthma, and women’s health conditions such as menstrual and menopausal symptoms. However, its natural pyrrolizidine alkaloid content poses safety concerns, making efficient purification essential for commercial applications.
Scaling Up Purification with Continuous CPC
To meet industrial-scale chromatography demands, we tested Continuous CPC on red clover extract containing elevated PA levels. Given the broad polarity range of both isoflavones and pyrrolizidine alkaloids, RotaChrom’s proprietary CPC platform proved highly effective for continuous purification. The process was performed in Multiple Dual-Mode (MDM) CPC, allowing periodic alternation between ascending and descending modes, dynamically switching the roles of the mobile and stationary phases in every cycle. This ensured continuous operation while maintaining high separation efficiency.
Results: 3x Productivity Increase Without Higher Operating Costs
A direct comparison between traditional batch chromatography and the implemented Continuous CPC process—as illustrated in the bar graphs—demonstrates the significant advantages of Continuous CPC:
✔ 3x higher productivity
✔ No significant increase in total solvent consumption, keeping OPEX stable
✔ Scalable industrial solution for continuous bio-manufacturing
By enabling high-efficiency, continuous purification, Continuous CPC is setting a new standard in industrial-scale chromatography, making large-scale extraction and remediation faster, more cost-effective, and more sustainable.
CPC offers many benefits compared to traditional chromatographic solutions.
HPLC, flash, supercritical fluid chromatography (SFC), simulated moving bed (SMB) and traditional chromatographic solutions are all demonstrably outperformed by CPC.
Without the need for silica gel or cartridges, and the benefit of easy, linear scalability, plus the solvent handling capacity demonstrably make CPC the most cost-efficient chromatography solution long-term.
CPC vs Flash Chromatography
No silica gel
One of the main expenses associated with flash chromatography is the cost of the separation column, which is typically filled with silica gel. Silica gel is a porous material that is used to adsorb and separate compounds based on their size and charge. It is a consumable material that needs to be replaced regularly, and the cost of the gel can add up over time.
No flash cartridges
Another factor to consider is the cost of the flash cartridge, which is the container that holds the separation column. Flash cartridges come in various sizes and configurations, and the cost can vary depending on the brand and the size of the cartridge. It’s important to choose a cartridge that is suitable for the volume of sample and the amount of solvent needed for the separation, as using a larger cartridge than necessary can increase the cost unnecessarily.
No risk of sample loss
CPC uses liquid-liquid chromatography, meaning there is little risk of sample loss or degradation due to interaction with the column, and the columns can be reused after rinsing, leading to less waste and lower expenses.
CPC vs High-Performance Liquid Chromatography (HPLC)
No resin, no irreversible binding
In contrast to CPC, HPLC utilizes solid-liquid partitioning that can lead to irreversible binding of contaminants on the columns and also sample loss. The resin makes HPLC an expensive technique compared to using solvents in both phases as in the case of CPC.
Hiqh selectivity for a variety of compounds
CPC offers high selectivity for compounds with similar polarities and is effective for sensitive and thermally labile compounds. Preparative HPLC provides high resolution and efficiency, particularly for compounds that interact well with the solid stationary phase.
One setup for a wide range of compounds
CPC can handle a wide range of compounds, including those that are difficult to separate by traditional chromatography methods. Preparative HPLC is versatile but may require different column chemistries and conditions to handle a diverse range of compounds.
Separation of complex mixtures
CPC is highly and easily scalable, often preferred for larger-scale separations of natural products and complex mixtures.
A variety of solvent systems for complex matrices
CPC is generally more solvent-efficient and can operate with a variety of solvent systems, potentially reducing costs. Preparative HPLC may require large volumes of high-purity solvents, which can increase operational costs.
CPC vs Simulated Moving Bed (SMB)
Separation of complex mixtures
While SMB allows the separation of 2 or 3 compounds of mixtures, CPC allows the separation of of multiple components from a complex crude mixture.
Liquid-liquid operation
By the liquid-liquid based operation of CPC, it excels in handling delicate compounds and achieving high selectivity for similar polarity compounds without the issues associated with solid stationary phases. Therefore, it provides high versatility for a wide range of applications. SMB operates with a series (4-8) of chromatographic HPLC columns, which can irreversibly bind contaminants and are also expensive.
Multiple-dual modes
While the theoretical true moving bed advanced technology could increase the productivity of SMB, two interconnected CPC systems (Continuous CPC) have been developed and can operate simultaneously in multiple-dual modes, allowing high loadability and throughput (thanks to continuous sample injection). The automated, cyclic operation of the two CPCs allows for high productivity and high purity.
Solvent purity is not an issue
In CPC, any grade purity of solvent can be used, while SMB needs HPLC grade solvents
Continuous CPC is superior for large-scale, continuous separations with high productivity and efficiency, making it ideal for industries requiring high-throughput processing.
Chiral separation
Separation of chiral compounds is traditionally done with SMB, however, RotaChrom has proven the capability of CPC in chiral purification with numerous successful case studies.
CPC vs Multicolumn Countercurrent Solid Gradient Purification (MCSGP)
Solvent Flexibility and Efficiency
Although MCSGP can utilize a range of solvents, it is still dependent on the solid stationary phases used in the columns. This can limit the range of solvents and conditions that can be effectively used without damaging the stationary phase or altering its properties. CPC does not rely on solid stationary phases, instead, it uses immiscible liquid phases. This allows for a broad range of solvent systems to be employed, making it highly adaptable to different types of solutes and solvents. This flexibility can result in more efficient separations and the ability to handle a wider variety of compounds, including those that are sensitive to solid phases.
Scalability and Throughput
While MCSGP is also scalable, the need for multiple columns and the complexity of their operation can make the scale-up process more challenging and costly. CPC systems are simple and can be scaled up easily from laboratory to industrial scale. The absence of solid stationary phases reduces the complexity and cost of scale-up processes.
Lower Risk of Column Fouling and Degradation
The use of solid stationary phases in MCSGP can lead to fouling, degradation, or changes in column performance over time, especially with complex or particulate-laden samples. This can result in increased maintenance costs and downtime. Since CPC uses liquid-liquid partitioning without solid phases, there is no risk of column fouling or degradation of stationary phases. This leads to more consistent performance over time and reduces the need for maintenance and replacement of components.
Environmental and Cost Benefits
MCSGP can consume more solvents, especially during the column regeneration phases, and generate more solid waste from the columns. This can increase both environmental impact and operational costs. CPC can be more environmentally friendly as it often requires less solvent overall, and the solvents used can be easily recovered and reused. The lack of solid phases also reduces the waste generated from spent columns.
Operational Simplicity
MCSGP requires careful coordination of multiple columns and precise control of solvent gradients and flow rates. This complexity can increase the time and expertise required for method development and routine operation. The operation of CPC systems is straightforward with fewer parameters to control compared to MCSGP. This can make method development and routine operation easier and faster.
CPC vs Supercritical Fluid Chromatography (SFC)
Quickly switch between different compounds
One of the main advantages of CPC is its ability to quickly switch between different compounds of interest (CoI). This is particularly useful for users who need to purify a large number of compounds, as it allows for efficient and flexible operation. In addition, CPC has a rapid setup and cycle time, which can further increase productivity and efficiency.
Applicable to a wide range of compounds
CPC provides high selectivity and efficiency for a broad range of compounds, including those with similar polarities. SFC offers high resolution and fast separations, especially for non-polar and moderately polar compounds.
Continuous CPC benefits
Both methods can handle large volumes of sample and they are highly scalable, but CPC in continuous operation works with lower solvent consumption due to its more controlled manner, making it also cost-effective.
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