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In UV-VIS, an isosbestic point is often interpreted as implying the occurrence of a single linearly independent reaction. The simplest examples of isosbestic points involve only two species, but isosbestic points do not imply the participation of only two species (e.g. the IUPAC example involves 5 species), which is a common misconception [1].
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.
Two other issues that must be considered in setting up an absorption spectroscopy experiment include the optics used to direct the radiation and the means of holding or containing the sample material (called a cuvette or cell). For most UV, visible, and NIR measurements the use of precision quartz cuvettes are necessary.
The absorbance measured by the Spectronic 20 is the sum of the absorbance of each of the constituents of the solution. Therefore, the Spectronic 20 can be used to analyze more complex solutions. For example, if a sample solution has two light-absorbing compounds in it, then the user performs measurements at two different wavelengths and ...
Hyperchromicity is the increase of absorbance (optical density) of a material. The most famous example is the hyperchromicity of DNA that occurs when the DNA duplex is denatured. [1] The UV absorption is increased when the two single DNA strands are being separated, either by heat or by addition of denaturant or by increasing the pH level.
An optical spectrometer (spectrophotometer, spectrograph or spectroscope) is an instrument used to measure properties of light over a specific portion of the electromagnetic spectrum, typically used in spectroscopic analysis to identify materials. [1]
This absorbance value can then be used to determine the concentration of a given element (or atoms) within the sample. The relationship between the concentration of atoms, the distance the light travels through the collection of atoms, and the portion of the light absorbed is given by the Beer–Lambert law .
Schematic of energy levels involved in two photons absorption. In atomic physics, two-photon absorption (TPA or 2PA), also called two-photon excitation or non-linear absorption, is the simultaneous absorption of two photons of identical or different frequencies in order to excite an atom or a molecule from one state (usually the ground state), via a virtual energy level, to a higher energy ...