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[6] The dilution method and concentration difference spectrophotometry: New designs for an old method

Methods in Enzymology, ISSN: 0076-6879, Vol: 61, Issue: C, Page: 65-96
1979
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Article Description

This chapter discusses the dilution method and concentration difference spectrophotometry. The dilution method involves the measurement of the concentrations of the associated or dissociated species or both, after each dilution step of solvent addition. The measurements can be made by light absorption, fluorescence intensity, circular dichroism, conductance, sedimentation techniques, and others, with the limits set only by the properties of the solutes and the sensitivities of the measuring equipment available. The method of concentration-difference spectra is used to show the way in which data acquisition and workup can be facilitated with a double-beam spectrophotometer employing reference and sample cuvettes. However, computers and highly stable spectrophotometers now eliminate the need for using the reference cuvette for difference spectra.

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