Impurities/degradants techniques

In chromatography techniques, selectivity can be proved by the existence of good separation between the analyte and the other components (such as the matrix, impurities, degradation product(s), and metabolites). A consequence of this requirement is that the resolution of the analyte from the other components should be more than 1.5-2.0. In order to detect the possibility of coelution of other substance(s), the purity of the analyte peak should also be determined. For instance, the UV-Vis spectrum of the analyte peak/spot can be used to determine 4the purity of the analyte peak/spot, in this case the correlation coefficient V (this term is used by the software of DAD System Manager Hitachi, and CATS from Camag). With the same meaning and mathematical equation, other terms are used, such as Match... 

In this chapter, we give an overview on how the API techniques work and which factors have an important influence on the performance. Examples are presented mostly from published work to demonstrate how LC-MS, LC-MS-MS, collision-induced dissociations (CIDs), accurate mass measurements and hydro-gen/deuterium exchange have been systematically and successfully applied in the structural elucidation of impurities, degradation products and metabolites. In addition, these also illustrate how mass spectrometry has offered a third dimension to chromatographic method development and validation. 

Selectivity is often referred to as the specificity of an analytical method and is a measure of the discriminating ability of the technique. The general requirement for specificity is that the method should be capable of unambiguously determining the compounds of interest in the presence of impurities, degradation products and other sample matrix components. A specificity study often involves accelerated degradation studies to ensure all degradation products will not interfere and the collection of likely process impurities. Often a placebo sample is assayed to check for interference fi om the sample matrix. 

Of the impurity/degradant enhancement techniques available, the phase-solubility analysis technique requires minimal sample handling from the... 

Impurity enhancement techniques such as fraction collection and phase equilibrium purification can be used to provide enriched samples for use in the method development process.23 When using the fraction collection approach, one or more cuts (fractions) of the chromatographic separation of a bulk lot or mother liquor are isolated. The excess solvent in these fractions is then evaporated to achieve the desired concentration enhancement. These fractions typically contain extraneous peaks because of the presence of salts in the mobile phase or sample degradation during the concentration step. The salts can be removed by extraction and/or a LC cleanup step. To insure that these extraneous peaks/artifacts are not identified as key peaks for separation, the original bulk lot or mother liquor should be included in the method development sample set. The same holds true for phase-equilibrium-purification supernatants. 

In addition to identifying structural units, degradative techniques can provide information about additives, impurities, residuals, structural defects, etc. However, in all determinations, it is absolutely necessary to maintain identical conditions so that the fragmentation of macromolecules will be reproducible. Pyrolytic methylation is also used for the identification of polymers [246]. 

Modern spectroscopy plays an important role in pharmaceutical analysis. Historically, spectroscopic techniques such as infrared (IR), nuclear magnetic resonance (NMR), and mass spectrometry (MS) were used primarily for characterization of drug substances and structure elucidation of synthetic impurities and degradation products. Because of the limitation in specificity (spectral and chemical interference) and sensitivity, spectroscopy alone has assumed a much less important role than chromatographic techniques in quantitative analytical applications. However, spectroscopy offers the significant advantages of simple sample preparation and expeditious operation. 

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