Abstract

The coherent body of research described in the existing published work is
concerned with new assay method development and validation using novel
systematic approaches for pharmaceutical and diagnostic compounds.
The first stage of the research was to study how analytical method development
and validation are typically carried out at present and to formulate this into a
simple step-by-step approach. Such a template and protocol was not only used
as the foundation of this research programme but could also serve as a simple
systematic guide for other practitioners and those new to the field. Furthermore,
it was recognised that this protocol should satisfy the requirements of the most
strategically important regulatory agencies.
The second stage of this research involved evaluation and application of the
above validation approach to new methods that were developed for a diverse
range of analytes and samples. A new purity assay for 1,10-phenanthroline-5,6-
dione and 4,7-phenanthroline-5,6-dione using high-performance liquid
chromatography (HPLC) was developed and validated. Impurities in these
compounds were identified by liquid chromatography-mass spectrometry (LCMS).
Best practice in method development and validation is equally important in
the analysis of both active components and excipients in formulated products. In
the first case, a liquid chromatography assay method for determining the content
of 2-(diethylamino)-N-(2,6-dimethylphenyl) acetamide in a gel formulation was
developed and validated. In the second case, the individual contents of three phydroxy
benzoic acid ester preservatives in a complex multi-component sample
were determined following the development and validation of a liquid
chromatography method.
Finally, the validation approach was evaluated as applied to another analytical
technique. Here, gas chromatography (GC) successfully used to develop a novel
assay for p-cymene in tea tree oil formulations presented different analytical
problems because of the very complex nature of this natural product. Stability
study information to increase the shelf life of the product and validation data for
the analytical method for p-cymene content was critically evaluated.
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In essence, the critical review of the requirements for method validation for
various agencies and the subsequent preparation of guidelines on how to go
about method validation have had a significant impact on how analytical
practitioners worldwide go about method development and, more importantly,
method validation. Further it was possible to apply these guidelines to conduct a
series of effective, successful method validation for assays involving a range of
typical pharmaceutical samples.