Purification of APIs

Pharmaceuticals (Synthetic)

Preparative chromatography has been widely used as a purification technique for pharmaceutical APIs. Purity, recovery (yield), throughput and cost are the drivers in this industry. Preparative chromatography provides customers with sufficient amounts of materials with the desired quality to initiate preclinical and clinical trials. This allows for alternative commercialization routes to be explored with no loss in development time.

The priority in purification depends on the development stage. In preclinical chemistry, the key is a quick method of development to have samples ready for initial activity tests. At this early stage, while time is a major factor, purification cost is not emphasized because the synthesis is carried out on a very small scale.

ZEOprep and ZEObeads

Flash chromatography is a widely used method to provide larger quantities of pre-purified material. This complimentary technique can also be used as a powerful tool for (non-chiral) pre-purification steps in the early development/preclinical phases of API manufacturing.

For the early API stage, Zeochem supplies efficient irregular shaped (ZEOprep) silica and microbead shaped (ZEObeads) silica in various pore and particle sizes, as well as modifications. When it comes to removing catalyst for API manufacturing steps, bonded irregular silica can be used as a reliable and cost-efficient scavenger.

ZEOsphere

During late clinical phases and in the commercialization phase when the active pharmaceutical ingredient (API) is required at large scale, the process needs to be carefully optimized to reduce the costs and maximize efficiency. Our ZEOsphere Classical silica provides all required reversed phases for the more complex preclinical, clinical and commercialization phases.

ZEOsphere DRP Mixed-Mode products can substantially reduce production costs of charged APIs. Higher recovery (yield) can be achieved from the improved selectivity, while also decreasing the organic solvent usage. Depending on the objective, a single-step purification is highly possible.


Applications

In today's competitive environment, synthetic chemists are required to deliver a large number of compounds for screening, while also focusing on cost reduction and speed to market. The purification process can often be a source of bottlenecks which has created a need for optimizing the process to meet the product quality requirements.

To optimize the process, normal-phase, reversed-phase and ion-exchange chromatography are frequently used as separation or purification techniques. Our ZEOsphere product line offers all required silica-based stationary phases by removing bottlenecks and guaranteeing the highest technical and economic efficiency for the purification processes.

Due to improved selectivity, our ZEOsphere DRP Mixed-Mode product line can decrease purification time and substantially increase yields. Working with ZEOsphere DRP products will also decrease your organic solvent consumption.

For the purification of small molecules, peptides (NCE or generics), insulin (analogues), proteins and oligonucleotides, it is often difficult to use methods similar to those applied in the purification of other compounds, mainly due to their complexity. Purification methods usually utilize various principles of chromatography, such as ion exchange chromatography and medium- or high-pressure reversed-phase chromatography.

Due to the increasing government and public pressure to decrease costs of medicines, the purification process should be as simple as possible and contain a minimum of steps. However, the reality is that by using today’s classical ion-exchange and or reversed-phase purification methods, multiple steps are still required to achieve the desired purity. Of course, this has a huge impact on recovery (yield). Also, a more organic solvent must be used — all resulting in an inefficient, costly purification process.

ZEOsphere DRP Mixed-Mode can substantially improve the selectivity of charged molecules from the adsorption on the hydrophobic sites and repulsion on the charged sites.

This enhanced selectivity leads to the desired purity more quickly and will considerably increase the recovery (yield). Because ZEOsphere DRP Mixed-Mode can work with low organic solvent conditions, a decrease in organic solvent usage is obtained.

ZEOsphere DRP Mixed-Mode is today’s answer to improving compound purification and considerably lowering production costs.

  • Generics
  • Insulin (analogues)
  • Peptides
  • Proteins
  • Oligonucleotides
  • Small Molecules

Custom Solutions

Contact Zeochem to discuss your current or future needs.
Zeochem's technical service team, engineers and chemists have decades of experience and are here to answer questions, troubleshoot and find solutions to your most challenging problems.

Related Products

Classical Normal Phase – Bare (Unbonded) Silica

Normal Phase Silica phases are suitable for separations of polar and basic organic compounds, such as vitamins, steroids, as well as pharmaceuticals.

Classical Normal Phase – CN

Normal Phase Silica phases are suitable for separations of polar and basic organic compounds, such as vitamins, steroids, as well as pharmaceuticals.

Classical Normal Phase – Diol

Normal Phase Silica phases are suitable for separations of polar and basic organic compounds, such as vitamins, steroids, as well as pharmaceuticals.

Classical Normal Phase – NH2

Normal Phase Silica phases are suitable for separations of polar and basic organic compounds, such as vitamins, steroids, as well as pharmaceuticals.

Classical Reversed Phase – C18

ZEOsphere C18 phases are the most preferred as they offer an excellent range of hydrophobic separation power along with high surface area coverage.

Classical Reversed Phase – C4

ZEOsphere C4 is commonly used to separate large macromolecules such as proteins

Classical Reversed Phase – C8

An excellent range of hydrophobic separation power along with high surface area coverage, but when less retention compared to a C18 is needed.

Classical Reversed Phase – CN

Common applications based on Cyano phases include the separation of flavonoids, extraction of polar compounds from non-polar samples as well as analytes with a wide range of hydrophobicity.

Classical Reversed Phase – Phenyl and Derivatives

Classic reversed phase material based on 100Å and 120Å Ultra-pure silica with high surface area. ZEOsphere Phenyl as reverse phase or in HILIC mode shows alternative selectivity to C18 and C8 column

ZEObeads 50 C18 / 20-30µm

Spheroidal shape alkyl-bonded silica used in reversed-phase chromatography, preferably where superior flow performance and selectivity are needed.

ZEObeads 50 C8 / 20-30µm

ZEObeads 50 C8 / 20-30um is a spheroidal shaped silica with a reduced alkyl chain length, slightly less hydrophobic in reverse phase chromatography. The shorter alkyl chain length allows in some cases a better interaction with the target compound.

ZEObeads 50 Diol / 20–30µm

ZEObeads 50 NH2 / 20-30um is a spheroidal shaped amine bonded silica. It is used in normal phase chromatography for target compounds with more alkaline pH and as an excellent metal scavenger under conditions of superior flow performance.

ZEObeads 50 NH2 / 20-30µm

ZEObeads 50 NH2 / 20-30um is a spheroidal shaped amine bonded silica. It is used in normal phase chromatography for target compounds with more alkaline pH and as an excellent metal scavenger under conditions of superior flow performance.

ZEOprep 60 C18 / 40-63µm

Alkyl-bonded silica for reversed-phase chromatography is a robust and reliable packing material, widely used in industrial separations as well as...

ZEOprep 60 C8 / 40–63µm

Alkyl-bonded silica with shorter chain length and slightly reduced hydrophobicity which is used in reversed-phase chromatography. The shorter alkyl length...

ZEOprep 60 CN / 40-63µm

Cyan-bonded silica is more polar than C18/8 bonding but is mostly used in reversed-phase chromatography for polar compounds and for...

ZEOprep 60 Diol / 40-63µm

Diol-bonded silica, more polar than C18/C8 types. Used in reversed- as well as in normal-phase chromatography, it is the product...

ZEOprep 60 NH2 / 40-63µm

Amine-bonded silica is mostly used in normal-phase chromatography and serves as an efficient metal scavenger in batch applications, best for...

ZEOprep 60 NH2P / 40–63µm

Primary, secondary amine-bonded silica and is more polar than C18/C8 phases. It is used in reversed-phase as well in normal-phase...

ZEOprep 60 SAX / 40-63µm

Silica-bonded for anion exchange chromatography.

ZEOprep 60 SH / 40-63µm

Thiol-bonded silica most efficiently serves as a metal scavenger, but it is also used as a means to remove palladium...

ZEOprep 90 C18 / 40-63µm

Alkyl-bonded silica with 90 angstrom pore size that has proved to be a robust packing material in particular for flash...

ZEOprep 90 C18 / 60-200µm

Alkyl-bonded silica with 90 angstrom pore size and a larger particle size distribution which provides a higher flow rate under...

ZEOprep 90 C8 / 40-63µm

Alkyl-bonded silica with 90 angstrom pore size with shorter alkyl chain length and slightly reduced hydrophobicity. It is used in...

ZEOprep 90 CN / 40–63µm

Cyan-bonded silica with 90 angstrom pore size, more polar than C18/8 bonding but mostly used in reversed-phase chromatography for polar...

ZEOprep 90 NH2 / 40-63µm

Amine-bonded silica with 90 angstrom pore size, mostly used in normal phase chromatography. It serves as an efficient metal scavenger...

ZEOprep Bonded Silica

Bonded ZEOprep silica is used in liquid chromatography, flash chromatography and solid phase extraction. It can be used as scavengers...

ZEOprep Unbonded Silica

Unbonded ZEOprep silica is mostly used as column packing material for preparative chromatography and the purification of natural products, food...


Frequently Asked Questions

  • The larger the molecular weight of the target molecule to be separated the larger the pore size needs to be. The sample molecules need to be able to interact with the pores of the particle to create an efficient separation mechanism. If the pores are smaller than your target molecule it will not be retained and elute immediately.
  • The larger the average pore size of the material the smaller will be the effective surface area. If you have less surface area available, your separation is generally less efficient. It is important to choose the right pore size for your target molecule.
  • Size and distribution of the silica particles effect the packing efficiency of the columns and therefore process performance.
  • The higher the packing density, the higher the surface area, the more interaction between eluent and separation material resulting in higher efficiency separation. However, the higher the packing density, the smaller the mean particle size is and therefore the higher the backpressure will be.

Bonding different chemical groups onto the surface of bare silica results in functionalized silicas which give higherselectivity towards specific molecules.

  • Bare/Silica has a high polar surface, it is the most popular stationary phase, ideal for conventional applications (normal phase separations, non-polar target molecules, pharmaceuticals, natural products)
  • C4; suited to molecules with large hydrophobic regions, peptides, proteins. Usually combined with a large pore size for big biomolecules (>1000Da)
  • PHE/phenyl; moderately nonpolar for aromatic compounds, aflatoxins, caffeine, phenols.
  • C8; highly hydrophobic pesticides, peptides, drugs
  • C18; more apolar than C8 and in reversed phase chromatography higher retention than C8. Has a non-polar surface, ideal for pharmaceuticals, steroids, fatty acids, peptides, proteins, pesticides, PCB’s
  • Amino/NH2has a medium polar surface, ideal for carbohydrates and nitrogen containing heterocycles and amines. Scavenger applications.
  • Diolhas a lesspolar surface (than bare silica) and still many hydroxy-groups, ideal for lipids; Our best option for many SFC applications.
  • Cyano/CN unique selectivity for polar analytes with widely different chemistry eg. Cyclosporine, carbohydrates
  • Thiol/SH; Mainly used for scavenging
  • SCX/SO4:Ion Exchange for strong cation
  • SAX/TMA; Ion exchange for weak anion
  • NH2P; Ion exchange for scavenging
  • Morphology of the silica particles; meaning bead shape
  • Surface area: this is a function of particle- and pore size
  • Surface functionalisation
  • Particle size distribution in terms of range and actual distribution of particles within the range (D10/D50/D90)
  • Irregular silica beads to be used in low -medium pressure systems
  • Spheroidal Silica beads can be used in low to high pressure systems
  • Spherical silica beads primarily used for high-resolution high-pressure systems