Jetzt verschenken. In den Warenkorb. Sie sind bereits eingeloggt. Klicken Sie auf 2. He has over fifteen years of academicresearch experience in Russia and fifteen years of research,teaching and consulting for major American pharmaceuticalcompanies, which has resulted in numerous papers and four bookchapters.
HPLC for Pharmaceutical Scientists
The main focus of Dr. Kazakevich's research is in HPLCretention mechanisms and the description of the analyte migrationthrough the column with an emphasis on the influence of adsorbentgeometry and surface chemistry. Note that acidic compounds will have comparatively higher retention and increased peak tailing on CSH columns. The HILIC mode, first developed by Alpert in s  , uses a hydrophilic stationary phase silica, diol, cyano, amide, ztwitterionic with an RPLC-like aqueous buffer and acetonitrile mobile phases, has enjoyed increasing popularity for the analysis of polar drugs, secondary drug metabolites, amino acids, peptides, neurotransmitters, oligosaccharides, carbohydrates, nucleotides, or nucleosides .
Improved versions of the highly successful coated polysaccharides chiral stationary phases CSPs were available in the late s. They brought similar versatility as the earlier coated CSPs but are more robust to aggressive solvents and can be used in normal phase, polarorganic and reversed-phase modes [5, 34].
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Wide-pore silica and polymeric packings first available in s were effective for separations of large biomolecules [1, 35]. With the advent of recombinant proteins as biopharmaceuticals such as monoclonal antibodies mAb , the need for detailed characterization by HPLC or capillary electrophoresis for QC purposes has become more pressing . Other high-impact developments in the past decade were in the HPLC detection and automation areas. The refractive index detector is not gradient compatible and does not have sufficient sensitivity .
Evaporative light scattering detection ELSD using nebulizer technology with laser light scattering detection is an option and is gradient compatible but has been recently surpassed by CAD uses nebulizer with corona discharge detection , which offers better sensitivity low ng and improved linearity. A common example is the stability-indicating or purity assay of active pharmaceutical ingredients API in which all impurities and degradants must be separated for accurate quantitation by UV detection.
Continued improvements have enhanced their capability and ease-of-use though they never appeared to be very popular [2, 6]. The latest market entry was an add-on software package compatible to two of the commonly used chromatography data systems. The software addresses the most time-consuming portion of the HPLC method development process optimization by automating method sequence from a user-defined design space using principles of Design of Experiments DoE and Quality by Design QbD .
This software can also perform statistical analysis and display optimum conditions after importing of the completed sequence results.
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Figure 5 illustrates the concepts and salient features of this software as implemented in our laboratory. Since concepts of DoE and QbD are well accepted, initial customer interest appeared to be high. AMDS is particularly useful to laboratories specializing in method development to support earlyphase drug development [2, 6].
For instance, to support process developments for API synthesis and drug product manufacturing from complex drug molecules with multiple chiral centers, as many as HPLC methods achiral and chiral methods for raw materials, starting materials, intermediates, final drug substances and drug products are typically developed in quick succession. Figure 5. Schematic diagram of an example instrument confi guration of an automated method development system using QbD software and a resulting graphics showing a design space with an operating region for optimum separations white color.
In summary, HPLC remains a highly dynamic field with numerous innovations in instruments, column technologies, and approaches in recent years. Pharmaceutical scientists are early adopters and beneficiaries of these newer technologies for research, development and quality control.
Newer column technologies allow faster and more efficient analysis of complex samples, chiral molecules and biomolecules. Finally, the rapid advancements of UHPLC and 2-D LC in combination with high-resolution MS have revolutionized life science research and promise to have even greater impact for clinical diagnostics. These developments are welcomed progress for the analytical chemist working in this rapidevolving world of drug development.
As a perspective for LC in the next years, some innovative works can be mentioned [42, 43]. This will almost certainly require the use of sub-mm bore capillary columns due to issues with heat generation and dissipation.
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These two research findings are unlikely to be ready for mainstream analytical work or QC very soon but the first results are encouraging and thus worth mentioning here. The authors thank Dr. Dawen Kou, Dr. Tom Walter, and Richard Verseput for their suggestions and comments on this manuscript.
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