p H will only significantly affect the retention of weak acids and bases.
A flow diagram of an HPLC system is illustrated in Figure 1.
For scientists and technicians new to method validation, this guide provides all the information and tools needed to develop a top-quality system.
For those experienced with method validation, the guide helps to upgrade and improve existing systems.
Note: CD-ROM/DVD and other supplementary materials are not included as part of e Book file. BLIESNER, Ph D, is the founder and President of Delphi Analytical Services, Inc., an organization dedicated to improving clients' level of compliance with Current Good Manufacturing Practices, Quality System Regulations, and Current Good Clinical Practice Regulations by providing training technology, training, and compliance products and services. Bliesner also offers lectures, short courses, and hands-on training on compliance issues for analytical laboratories.
He is the author of Establishing a CGMP Laboratory Audit System, also available from Wiley.
Moreover, the guide features detailed flowcharts and checklists that lead readers through every stage of method validation to ensure success.
All of the templates are also included on a CD-ROM, enabling readers to easily work with and customize them.
The following must be considered when developing an HPLC method: Mobile phase composition, for example, is the most powerful way of optimizing selectivity whereas temperature has a minor effect and would only achieve small selectivity changes.
Note: CD-ROM/DVD and other supplementary materials are not included as part of e Book file.' / brings order and Current Good Manufacturing Practices to the often chaotic process of chromatographic method validation.
The wide variety of equipment, columns, eluent and operational parameters involved makes high performance liquid chromatography (HPLC) method development seem complex.
Reverse phase is the choice for the majority of samples, but if acidic or basic analytes are present then reverse phase ion suppression (for weak acids or bases) or reverse phase ion pairing (for strong acids or bases) should be used. For low/medium polarity analytes, normal phase HPLC is a potential candidate, particularly if the separation of isomers is required.
Cyano-bonded phases are easier to work with than plain silica for normal phase separations.