Life sciences applications, HPLC

Most of the hplc instrumentation now in use is unsuitable for small bore columns. At the moment, the technique is used mainly in the applications laboratories of some instrument manufacturers (they are interested in selling it ). The method is potentially attractive in areas where sample sizes are very limited, for example in biochemical or life sciences applications, but whether or not it becomes widely accepted remains to be seen. 

Column diameter is an important parameter to consider in life science applications in which sample amounts are very limited and the components of interest may not be abundant. Researchers have reviewed micro HPLC instrumentation and its advantages.910 Nano LC-MS offers 1000- to 34,000-time reductions in the dilution of a sample molecular zone eluted from nano LC columns of 25 to 150 [Mi IDs in comparison to a 4.6 mm ID column. This represents a large enhancement of ion counts in comparison to counts obtained for the same amount of sample injected into a conventional 4.6 mm column. Solvent consumption for an analysis run or sample amount required for injection in a nano LC application may be reduced 1000 to 34,000 times compared to amounts required by an analytical column operated at a 1 mL/min flow rate. 

This chapter describes the use of HPLC in food, environmental, chemical (polymer, ion chromatography), and life sciences applications. The HPLC advantages, method requirements, and trends of these application areas are summarized with descriptions of the methodologies of key analytes. References for further studies are listed. 

HPLC life science applications focus on the separation, quantitation, and purification of biomolecules such as proteins, peptides, amino acids, nucleic acids, nucleotides, and polymerase chain reaction (PCR) amplification products.31 34 These are diversified and active research areas in medical research and drug discovery. 

Planar Chromatography in the Life Sciences. Edited by Joseph C. Touchstone Fluorometric Analysis in Biomedical Chemistry Trends and Techniques Including HPLC Applications. By Norio Ichinose, George Schwedt, Frank Michael Schnepel, and Kyoko Adochi... 

Increasingly, investigators in the life sciences have expressed interest in the application of HPLC to the assay of enzymatic activities. This method not only provides a method to enhance the separation of reaction components, it also allows extensive and complete analysis of the components in the reaction mixture during the reaction. In addition, it can employ sensitive detectors, and it can be used for purification. 

It is the ability of hplc to accomplish separations completely and rapidly that led to its original application to problems in the life sciences, particularly those related to purification. An analysis of the literature revealed that this technique was used primarily for the purification of small molecules, macromolecules such as peptides and proteins, and more recently antibodies. This application to purification has all but dominated the use of the method, and there has been a plethora of books, symposia, and conferences on the use of hplc for these purposes. However, it was only a matter of time before others began to look beyond and to explore the possibilities that result from the capacity to make separations quickly and efficiently. 

HPLC is a versatile technique applicable to diversified analytes, including labile molecules, ions, organic, and biopolymers. This chapter provides an overview of HPLC applications for the analysis of food, environmental, chemical, polymer, ion-chromatography, and life science samples. In food analysis, HPLC is widely used in product research, quality control, nutritional labeling, and residual testing of contaminants. In environmental testing, HPLC is excellent for the sensitive and specific detection of labile and nonvolatile pollutants... 

Most chromatographers have a good understanding of the basis and application of isocratic HPLC. Workers in the life sciences have adapted these principles to the special requirements of their individual samples, as described elsewhere in this book. Separations that use gradient elution, however, are inherently more complicated. All the variables present in isocratic elution... 

To date, HPLC has become the dominating chromatographic technique, with capillary GC being second only to it (for the more volatile analytes). Both GC and H PLC are mature separation techniques today however, HPLC is stiH being developed toward faster and more efficient separations and also partially toward miniaturized columns, particularly for applications in the life science area. The majority of the other techniques already mentioned are niche techniques today, but still important for a relatively smaller number of users compared to HPLC and GC. Electric potential-driven techniques have an added opportunity for new technolc y on microchips. 

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