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the pharmaceutical industry to predict the ability of candidate drugs to be absorbed

and distributed since they are good models of cell membranes. The technique uses a

phospholipid-coated filter disc and is configured in 96-well format with an ultraviolet

plate reader as a detection system.

11.5 Partition chromatography

11.5.1 Principle

Like other forms of chromatography, partition chromatography is based on differ-

ences in retention factor,k, and distribution coefficients,Kd, of the analytes using

a b

c

d

e

f

g

a

b

c

d

f

g

a

b+

c

d

e

f

g

0

0

12 24 36 48

0

50

100

%B

SynChropak PROPYL

0.256

0.512

0

0.256

0.512

0.768

0

0.256

0.512

0

50

100

%B

PolyETHYL A

0

50

100

%B

PolyPROPYL A

Absorbance at 220 nm

Time (min)

Fig. 11.8Chromatogram of a mixture of proteins separated by hydrophobic interaction chromatography using

different stationary phases. A linear gradient elution program was used changing from 0 to 100% mobile phase B

in 40 min. Mobile phase A: 1.8 M ammonium sulphateþ0.1 M potassium phosphate pH 7.0. Mobile phase B:

0.1 M potassium phosphate pH 7.0. Elution was monitored at 220 nm. (Reproduced with permission from

K. Benedek (2003) High-performance interaction chromatography, inHPLC of Peptides and Proteins: Methods

and Protocols, Methods in Molecular Biology No. 251, M.-I. Aguilar (ed.), Humana Press, Totowa, NJ.)

455 11.5 Partition chromatography