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The concentration of sites is given by dividing the total number of sites (S 0) covering the whole surface by the area of the adsorbent (a): [ S 0 ] = S 0 / a . {\displaystyle [S_{0}]=S_{0}/a.} We can then calculate the concentration of all sites by summing the concentration of free sites [ S ] and occupied sites:
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.
The absorbance of a material that has only one absorbing species also depends on the pathlength and the concentration of the species, according to the Beer–Lambert law =, where ε is the molar absorption coefficient of that material; c is the molar concentration of those species; ℓ is the path length.
The absorption coefficient is fundamentally the product of a quantity of absorbers per unit volume, [cm −3], times an efficiency of absorption (area/absorber, [cm 2]). Several sources [ 2 ] [ 12 ] [ 3 ] replace nσ λ with k λ r , where k λ is the absorption coefficient per unit density and r is the density of the gas.
The amount concentration c is then given by = (). For a more complicated example, consider a mixture in solution containing two species at amount concentrations c 1 and c 2 . The decadic attenuation coefficient at any wavelength λ is, given by μ 10 ( λ ) = ε 1 ( λ ) c 1 + ε 2 ( λ ) c 2 . {\displaystyle \mu _{10}(\lambda )=\varepsilon _{1 ...
The blank solution should be the same pH and of a similar ionic strength as the sample solution. Example: using water for the blank measurement for samples dissolved in TE may result in low 260/230 ratios. A260/A280 Residual phenol or other reagent associated with the extraction protocol. A very low concentration (< 10 ng/μL) of nucleic acid.
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]
The absorption coefficient of a volume, denoted μ a, and the scattering coefficient of a volume, denoted μ s, are defined the same way as the attenuation coefficient. [ 6 ] The attenuation coefficient of a volume is the sum of absorption coefficient and scattering coefficients: [ 6 ]