ISOLUTE® PE-AX/SCX-2

1. Cartridge equilibration (can be optional)
2. Sample loading
3. Cartridge washing (impurity elution, product is retained)
4. Product elution

It may also be possible to retain impurities in step 3, such that the cartridge becomes a one pass, one step purification system.

1. Equilibration: Pipette 10 mL of DCM, MeOH or THF into cartridge and allow to drain.
2. Loading: Pipette approximately 1 mL of sample standard onto the top of the column and allow to flow under gravity onto the column until the sample falls below the top frit surface. Wash column with one volume of DCM. Note: Elution will contain neutral component.
3. Acidic Compound Elution (Step 1): Elute compound with 1M HCl solution, and wash with additional equilibration solvent if desired, collecting released sample in appropriate container. Note: Elution will contain acidic component.
4. Basic Compound Elution (Step 2): Elute compound with 10 mL of 2M ammonia solution, and wash with additional equilibration solvent if desired, collecting released sample
   in appropriate container. Note: Elution will contain basic component.

Further Notes: equilibration, loading and elution solvents may be substituted. Base cartridge loading calculations on approx. 60% cartridge capacity for initial run, depending
on the product, loading may be increased further following optimization.