Product analysis - Pharmaceutical Biotechnology: Concepts and Aapplications - page 185

7.4.2 High-performance liquid chromatography

HPLC occupies a central analytical role in assessing the purity of low molecular mass pharma-
ceutical substances (Figure 7.3). It also plays an increasingly important role in analysis of mac-
romolecules, such as proteins. Most of the chromatographic strategies used to separate proteins
under 'low pressure' (e.g. gel fi ltration, ion exchange, etc.) can be adapted to operate under high
pressure. Reverse-phase-, size-exclusion- and, to a lesser extent, ion-exchange-based HPLC chro-
matography systems are now used in the analysis of a range of biopharmaceutical preparations.
On-line detectors (usually a UV monitor set at 220 or 280 nm) allows automated detection and
quantifi cation of eluting bands.
HPLC is characterized by a number of features that render it an attractive analytical tool. These
include:

• excellent fractionation speeds (often just minutes per sample);

• superior peak resolution;

Figure 7.3 Photograph of a typical HPLC system (the Hewlett Packard HP1100 system). Photograph courtesy
of Hewlett Packard GmbH, Germany