The Application Notebook-10-02-2009

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The Application Notebook

Development and production of biopharmaceuticals is a growing segment of the pharmaceutical industry. The development of vaccines is only one - but as a result of the current H1N1 flu hysteria it is the most striking - segment of this emerging field. Recombinant proteins, ranging from insulin products to therapeutic antibodies, represent another important group of biopharmaceuticals. With the introduction of the first so-called biosimilars in Europe the demand for highly efficient analysis methods is further increasing. Newly developed stationary phases for the typical modes of biochromatography of native proteins such as SEC or IEC open up new opportunities for increasing throughput in the bioanalytical lab.

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The Application Notebook

The analysis of the molar mass distribution of polyethylene and polypropylene resins by GPC/SEC has always been considered a demanding task because of the requirement of high temperature operation for dissolution and complex hardware design, which often results in high maintenance cost, in particular related to the autosampler/injector and detector units, and in other problematic and consuming tasks such as solvent handling added to column fragility, sample degradation or detector sensitivity–stability.

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The Application Notebook

Over-expression of recombinant proteins is commonly used for the production of protein reagents in industry and academia. Problems often occur relating to the stress put on the cells to deal with this huge increase in synthesis. Cellular proteins that are part of the protein synthesis machinery are often up-regulated under such conditions. Large quantities of the recombinant protein can be bound to these cellular proteins, making purification difficult.

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The Application Notebook

A critical part of research and quality control analysis of proteins involves verifying the primary and secondary structure of a protein. Peptide mapping is the main technique used to determine the structure of a protein as well as identify any post-translational modifications.

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The Application Notebook

As a result of their high toxicity and carcinogenicity, aflatoxins are of major concern for food producers, the food processing industry and consumers. Most countries have legislation setting maximum permissible limits for aflatoxins, which are in the low micrograms per kilogram for food matrices.

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The Application Notebook

"Amino acid" is a generic term referring to organic chemistry compounds that have amino and carboxyl groups. There are many known types of amino acids with natural origin. In addition to being the basic unit comprising proteins (one of the main constituents of living organisms), they are the raw material necessary for synthesis of physiologically active low-weight molecules such as neurotransmitters. Even as individual units they possess various types of physiological activity. They are researched widely in the fields of medicine and food. Recently, they are being widely used in health foods such as supplements.

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The Application Notebook

Recently a newly developed Kinetex 2.6 µm core-shell chromatographic particle has been commercialized that offers the performance benefits of sub-2 µm fully-porous particles but at substantially lower operating pressures.

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The Application Notebook

In this application, we demonstrate the use of supported liquid extraction (SLE) for the extraction of beta blockers and NSAIDS from plasma compared with traditional liquid–liquid extraction. SLE was demonstrated to yield consistent LOQs using lower sample volumes.

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The Application Notebook

The flexibility to use short sub-2 micron columns on UHPLC instruments at high flow-rates permits fast analysis with high resolution of complex samples. An Agilent ZORBAX Rapid Resolution High Definition (RRHD) Eclipse Plus C18 1.8 µm column separated seven different biocides in less than 1 minute with high resolution. A flow-rate of 1.7 mL/min was used on 2.1 × 50 mm column to achieve the rapid separation. The Agilent 1290 Infinity LC system was used as the column pressure just exceeded 1000 bar at this high flow-rate. A sample of hand sanitizer was found to contain 2-phenoxyethanol and methylparaben using the method.