ChromTech Chiral columns

　

　

-- ChromTech Chiral Column Selection Guide --

Column

Column Descriptions 

Column Applicability 
(type of samples)

Chiral-AGP

• Broadest applicability

• Acids, bases, and neutrals

• No derivatization

• The USP L41 column

  CHIRAL-AGP is the second generation chiral separation column based on the use of α1 -acid glycoprotein (AGP) as the chiral stationary phase. Changing the mobile phase composition, i.e. the pH, the concentration or the nature of the organic modifier, can regulate Enantioselectivity and retention. The column temperature also affects these parameters.

Extremely broad applicability. Most likely the column with the broadest applicability of all chiral columns. Separates all kinds or compounds:

• Amines (primary, secondary, tertiary and quaternary)

• Acids (strong and weak)

• Nonprotolytes (amides, esters, alcohols)

Chiral-HSA

• Acids and neutrals

• Excellent for hydrophilic acids

The chiral selector in this stationary phase is human serum albumin (HSA). Enantiomers of preferentially acidic compounds can be resolved directly, without derivatization. The column is operated in the reversed phase mode.

With the Chiral HSA column, both racemic acids and amino acids.

More narrow applicability than CHIRAL-AGP. Separates compounds containing one or more basic nitrogens together with one or more hydrogen accepting or hydrogen donating groups (alcohol, phenol, carbonyl, amide, ether, sulphoxide, ester, etc.).

Chiral-CBH

• Basic compounds

• Excellent for hydrophilic amines

 

Cellobiohydrolase (CBH) is a stable enzyme. This is also a reversed phase column, used for the direct separation of enantiomers. The column is preferentially used for the separation of the enantiomers of basic drugs from many compound classes.

More narrow applicability than CHIRAL-AGP. Separates preferentially weak and strong acids, zwitterionic and non-protolytic compounds.

 

FIRST CHOICE = CHIRAL-AGP

The columns overlap for some types of compounds; basic compounds can be separated on both CHIRAL-AGP and CHIRAL-CBH, acidic and neutral compounds can be separated on both CHIRAL-AGP and CHIRAL-HSA. However, as CHIRAL-AGP is a column with an extremely broad applicability, this column should be chosen first, if the analyte has not been separated on any of the columns. There are, however, some types of compounds where one of the other columns may be the first choice:

CHIRAL-HSA: very hydrophilic acids

CHIRAL-CBH: very hydrophilic amines / amino alcohols

 

METHOD DEVELOPMENT

The columns described above are all reversed-phase columns, giving many possibilities to affect both the retention and the enantioselectivity. The solutes are retained by three types of forces; ionic bonding (charged solutes), hydrophobic interaction and hydrogen bonding. The relative contribution of the different forces to the retention of the solutes, depending of the nature of the analyte. Analytes containing charged groups, hydrogen bonding groups and hydrophobic parts can be retained by interaction with corresponding groups on the chiral selector. From this follows that a separation can be affected by:

- pH                                          
- organic modifier concentration

- nature of organic modifier         
- buffer concentration

- type of buffer

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