Chiral   HPLC

Applications

Chromtech - Chiral Handbook 40p PDF

What is it!
Separation of Optical isomers by HPLC.
Often the critical moities in biologically active materials whether in bioscience
or pharmaceuticals.
This complex “mirror image” molecular separation is often given scant recognition
by researchers primarily due to the difficulty in separating and analysing such
raecemic mixtures.
Let alone the difficulties of separating and purifing on a Preparative scale.

Chromtech chiral HPLC-columns for direct resolution
of enantiomers
 - comprise 3 basic types

• CHIRAL-AGP ( USP chiral column L41 )
  The CHIRAL-AGP column has extremely broad applicability.
  As can be seen from the applications a very large number of basic, acidic and
  nonprotolytic drugs and related compounds have been separated into enantiomers.
  The columns resolve chiral compounds containing primary-, secondary- or tertiary amino
  groups or quarternary ammonium groups, as well as chiral compounds containing
  carboxylic acid functions or weaker acidic groups as well as chiral nonprotolytes
   like alcohols, amides, sulphoxides, esters etc.

• CHIRAL-CBH
  Cellobiohydrolase (CBH) is a very stable enzyme, which has been immobilized onto
   5 micrometer spherical silica particles. This is a reversed-phase column, used for
   the direct separation of enantiomers. The column is preferantially used for the separation
  of enantiomers of basic drugs from many compound classes.
   CHIRAL-CBH separates preferentially compounds containing one or more basic nitrogen
   together with one or more hydrogen accepting or hydrogen donating groups (alcohol, phenol, carbonyl, amide, ether, sulphoxide, ester etc.)

• CHIRAL-HSA
  The CHIRAL-HSA column is based on human serum albumin (HSA).
   The protein has been immobilized onto spherical 5 micrometer particles.
  Enantiomers of preferentially acidic compounds can be resolved directly, without derivatization.
  CHIRAL-HSA is especially suited for the resolution of weak and strong acids, zwitterionic
  and nonprotolytic compounds.
  However, basic compounds have also been resolved.

Extensive Application Notes are available . . . hundreds of separations often with huge “alpha” offering opportunies for
massive scale up in column load capacities

Application Areas
At pharmaceutical companies, hospitals, universities and chemical industry CHIRAL-CBH is used for the analysis of
enantiomeric purity and for bioanalysis.
An interesting and fast growing area is isolation of pure enantiomers on semipreparative columns.

Mobile Phases
The mobile phases are mixtures of phosphate or acetate buffers and organic solvents as 2-propanol or
acetonitrile containing 50mM disodium EDTA. The retention and the enantioselectivity can be regulated by changes in pH,
 buffer concentration and organic modifier (nature and concentration).

Chiral columns for LC/MS

Column Dimensions

• Inner diameters 2, 3, 4, and 10 mm

• Lengths of 50, 100 and 150 mm.

• Guards with length of 10 mm and the inner diameters 2, 3 and 4 mm are also available

Complete Chiral Column List

Column Type

CHIRAL-

-DIM (mm)

Analytical

AGP
CBH
HSA

150.4
100.4
50.4
150.3
100.
50.3

Micro

AGP
CBH
HSA

150.2
100.2
50.2

Semiprep

AGP
CBH
HSA

150.10
100.10

Analytical Guard 2/pk

AGP
CBH
HSA

10.42
10.32
10.22

Guard Column Holder

for All

CH10.3

Guard Column Connector

CON2

Micro GC Connector

CON4

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