By A. Fallon, R.F.G. Booth and L.D. Bell (Eds.)
This booklet is meant to familiarize biochemists with HPLC. Theoretical points of every mode of chromatography are mentioned in chapters 1-9, supplying an knowing of many of the modes of chromatography that are now attainable utilizing commercially to be had columns, from reversed section to affinity. useful points and instrumentation are coated in bankruptcy 10. the majority of the ebook, which follows, provides examples and functions of every mode of chromatography in present biochemical perform.
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Additional info for Applications of HPLC in Biochemistry
The increased sensitivity of fluorescence detectors means that the presence of contaminating material in the mobile phase can become a major problem. One method of circumventing this is to distill the solvents over a fluorescent reagent prior to use (Stein and Brink, 1980). 3. Refractive index (RI) detection Refractive index detection is the second most popular non-destructive method used in HPLC. RI detection utilises the principle of the change in direction of a beam of light as it passes through different matrices.
G ) Passivate the HPLC system regularly. Ch. 8. Fraction collectors A variety of fraction collectors are available from a number of manufacturers and include those which are able to collect low microlitre volumes in addition to those used in preparative HPLC which can collect much larger volumes. Fraction collectors which are to be used with HPLC systems require very rapid tube changing facilities and should also be resistant to the solvents used with HPLC systems; thus, fraction collectors which are designed for use with low pressure chromatography are usually inadequate.
For routine operations, such as in quality control monitoring, a simple UV photometer is usually adequate; however, for research work where compounds may need to be monitored at various wavelengths an alternative is to use a variable wavelength UV detector (VWD). These detectors are of two types; firstly, those which use a visible or UV source (tungsten lamp and deuterium lamp, respectively) in combination with a monochromator to select the desired wavelength, and secondly, those which use a photodiode array to allow monitoring either at a fixed number of wavelengths or across a whole spectrum of wavelengths.
Applications of HPLC in Biochemistry by A. Fallon, R.F.G. Booth and L.D. Bell (Eds.)