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Absorbance is defined as "the logarithm of the ratio of incident to transmitted radiant power through a sample (excluding the effects on cell walls)". [1] Alternatively, for samples which scatter light, absorbance may be defined as "the negative logarithm of one minus absorptance, as measured on a uniform sample". [2]
Spectrophotometry is a branch of electromagnetic spectroscopy concerned with the quantitative measurement of the reflection or transmission properties of a material as a function of wavelength. [2] Spectrophotometry uses photometers , known as spectrophotometers, that can measure the intensity of a light beam at different wavelengths.
In chemistry, the molar absorption coefficient or molar attenuation coefficient (ε) [1] is a measurement of how strongly a chemical species absorbs, and thereby attenuates, light at a given wavelength. It is an intrinsic property of the species.
For spectroscopy, it is generally desirable for a source to cover a broad swath of wavelengths in order to measure a broad region of the absorption spectrum. Some sources inherently emit a broad spectrum. Examples of these include globars or other black body sources in the infrared, mercury lamps in the visible and ultraviolet, and X-ray tubes.
Variable pathlength absorption spectroscopy uses a determined slope to calculate concentration. As stated above this is a product of the molar absorptivity and the concentration. Since the actual absorbance value is taken at many data points at equal intervals, background subtraction is generally unnecessary.
Parameters of interest, besides the wavelength of measurement, are absorbance (A) or transmittance (%T) or reflectance (%R), and its change with time. [4] [5] A UV-Vis spectrophotometer is an analytical instrument that measures the amount of ultraviolet (UV) and visible light that is absorbed by a sample. It is a widely used technique in ...
When an isosbestic plot is constructed by the superposition of the absorption spectra of two species (whether by using molar absorptivity for the representation, or by using absorbance and keeping the same molar concentration for both species), the isosbestic point corresponds to a wavelength at which these spectra cross each other.
The decadic absorbance of a scattering sample is defined as −log 10 (R+T) or −log 10 (1−A). For a non scattering sample, R = 0, and the expression becomes −log 10 T or log( 1 / T ), which is more familiar. In a non-scattering sample, the absorbance has the property that the numerical value is proportional to sample thickness.