Mixed-Mode Method Development Kits

Method development kits below can be used to develop methods for all your compounds. Initial experiments on 50 mm columns can be scaled up on 150 mm columns in Screen and Resolve HILIC and RP Mixed-Mode Kits.

The efficiency of HPLC method development depends both on the skill level of analysts as well as the tools their R&D lab provides to conduct a cost-effective method development. We are offering comprehensive method development kits which will allow you to do fast method development and cover 95% of applications. 

You can create any combination of our columns and create your own kit, or use one of the kits we recommend for you. If you are not familiar with method development in mixed-mode chromatography we suggest that you start with short columns. They will allow you to learn how to control retention time, elution order and peak shape. Using 3.0×50 mm columns can speed up your method development since you can run them at higher flow rates without consuming too much solvent, or creating high back pressure. All our columns are compatible with 100% ACN and 100% water and can run any type of gradient. You can use isocratic conditions as well as double and triple gradients. Mixed-mode columns allow you to convert your long gradient method to a much shorter isocratic gradient without sacrificing resolution between compounds in the mixture. Our mixed-mode columns let you retain and separate compounds based on their hydrophobic/hydrophilic properties, as well as ionic properties of the analyte and mixed-mode stationary phases. Control of different mechanisms of interaction can be done independently and will allow you to change the order of elution for compounds with different hydrophobic and ionic properties. 

Retention time for compounds of different nature can be controlled by changing the composition of the mobile phase. Amount of ACN, buffer pH, buffer concentration and buffer nature can be changed to successfully retain and separate compounds with different properties in one run without the use of ion-pairing reagents. Our columns have various ion-pairing reagents attached to the surface of silica gel.

 

Compound Type Mechanisms of Interaction How to Control Retention Recommended Column
Hydrophilic basic  compound Weak reversed-phase, HILIC, cation-exchange Low organic in RP mode, high organic in HILIC, buffer concentration and buffer pH 1st tier: Coresep 100, Amaze HD

2nd tier: Amaze TH, Amaze TR, Amaze SA, Heritage N

Hydrophilic acidic compound Weak reversed-phase, HILIC, anion-exchange Low organic in RP mode, high organic in HILIC, buffer concentration and buffer pH 1st tier: Heritage MA, Coresep SB, Amaze TH

2nd tier: Amaze HD, Amaze HA, Amaze TR

Hydrophilic neutral compound Weak reversed-phase, HILIC Low organic in RP mode, high organic in HILIC mode, buffer concentration and buffer pH 1st tier: Amaze TH, Amaze HD

2nd tier: Coresep 100, Heritage MA, Heritage N

Hydrophilic zwitterionic compound Weak reversed-phase, HILIC, cation- or anion exchange Low organic in RP, high organic in HILIC, buffer concentration and buffer pH 1 st tier: Coresep 100, Amaze HD

2nd tier: Amaze TH, Amaze SA, Heritage N

Hydrophobic basic compound Reversed-phase, cation-exchange Amount of ACN, buffer concentration and buffer pH 1st tier: Heritage MA, Coresep SB

2nd tier: Coresep 100, Amaze TH, Amaze HD, Heritage N

Hydrophobic acidic compound Reversed-phase, anion-exchange Amount of ACN, buffer concentration and pH 1st tier: Heritage MA, Coresep SB

2nd tier: Amaze HA, Amaze TH, Amaze HD

Hydrophobic neutral compound Reversed-phase Amount of ACN 1st tier: Heritage MA, Coresep 100

2nd tier: Coresep SB, Amaze TR, Amaze SA, Amaze HA

Hydrophobic zwitterionic compound Reversed-phase, cation-exchange, anion-exchange Amount of ACN, buffer concentration and pH 1st tier: Amaze TR, Heritage MC

2nd tier: Coresep SB, Amaze SA, Amaze TH, Amaze HD, Heritage N

Acidic polymer  Reversed-phase, HILIC, anion-exclusion Amount of ACN, buffer concentration and pH 1st tier: Amaze HD, Heritage N

2nd tier: Heritage MC

Basic polymer Reversed-phase, HILIC, cation-exclusion Amount of ACN, buffer concentration and pH 1st tier: Heritage MA

2nd tier: Coresep SB, Amaze HA