Advantages of Reversed phase for Preparative Chiral Separations

By: Jennifer E. Claus, Reporter US Volume 27.5

Astec CHIROBIOTIC columns operate in LC-MS- and prep-compatible reversed phase mode

The use of reversed phase mode in preparative separations provides a number of benefits over normal phase methods. In addition to eliminating solubility issues often experienced in nonpolar normal phase solvents, the utilization of reversed phase chromatography uses less toxic solvents than those associated with normal phase and provides timely sample recovery.

Unlike many cellulosic phases that are incompatible with reversed phase modes, Astec CHIROBIOTIC® phases possess multiple bonded chiral selectors, making them extremely robust and compatible with all commonly used HPLC solvents.1 The successful preparative separation of fluoxetine performed on a CHIROBIOTIC V2 in reversed phase mode is illustrated in Figure 1. For rapid sample recovery following purification, fractions of the separated enantiomers may be run through a VersaPak® C18 flash cartridge. The analytes adhere to the cartridge and are washed with water to remove unwanted buffer salts and other mobile phase additives. This washing procedure ensures the additives do not remain as contaminants in the pure final product. Each enantiomer may ultimately be eluted from the flash cartridge using a minimal amount of a polar organic solvent such as methanol or acetonitrile (100 mL or less). Removal of the organic solvent, may be done quickly under vacuum, yielding the dried separated enantiomers.

Figure 1 Chiral Preparative Separation of Fluoxetine on Astec CHIROBIOTIC V2 in reversed phase Mode (referenced 15044AST)

Reversed phase chromatography provides better solubility for polar analytes, uses nontoxic solvents, offers a method for removal of contaminants and mobile phase additives, and gives timely sample recovery with little solvent evaporation. In conclusion, the use of reversed phase mode for preparative chiral separations is a safe and efficient way to execute successful chiral preparative separations.

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  1. CHIROBIOTIC Handbook (T408131) (Available from Supelco.)


Trademark Attributions:

  • CHIROBIOTIC is a registered trademark of Sigma-Aldrich Co. LLC
  • VersaPak is a registered trademark of Sigma-Aldrich Co. LLC


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