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Ultraviolet and visible (UV-Vis) absorption spectroscopy is the measurement of the attenuation of a beam of light after it passes through a sample or after reflection from a sample surface. Absorption measurements can be at a single wavelength or over an extended spectral range. Ultraviolet and visible light are energetic enough to promote outer electrons to higher energy levels, and UV-Vis spectroscopy is usually applied to molecules or inorganic complexes in solution. The UV-Vis spectra have broad features that are of limited use for sample identification but are very useful for quantitative measurements. The concentration of an analyte in solution can be determined by measuring the absorbance at some wavelength and applying the Beer-Lambert Law.
Since the UV-Vis range spans the range of human visual acuity of approximately 400 - 750 nm, UV-Vis spectroscopy is useful to characterize the absorption, transmission, and reflectivity of a variety of technologically important materials, such as pigments, coatings, windows, and filters. This more qualitative application usually requires recording at least a portion of the UV-Vis spectrum for characterization of the optical or electronic properties of materials.
The UV-Vis spectral range is approximately 190 to 900 nm, as defined by the working range of typical commercial UV-Vis spectrophotometers. The short-wavelength limit for simple UV-Vis spectrometers is the absorption of ultraviolet wavelengths less than 180 nm by atmospheric gases. Purging a spectrometer with nitrogen gas extends this limit to 175 nm. Working beyond 175 nm requires a vacuum spectrometer and a suitable UV light source. The long-wavelength limit is usually determined by the wavelength response of the detector in the spectrometer. High-end commercial UV-Vis spectrophotometers extend the measurable spectral range into the NIR region as far as 3300 nm.
The light source is usually a deuterium discharge lamp for UV measurements and a tungsten-halogen lamp for visible and NIR measurements. The instruments automatically swap lamps when scanning between the UV and visible regions. The wavelengths of these continuous light sources are typically dispersed by a holographic grating in a single or double monochromator or spectrograph. The spectral bandpass is then determined by the monochromator slit width or by the array-element width in array-detector spectrometers. Spectrometer designs and optical components are optimized to reject stray light, which is one of the limiting factors in quantitative absorbance measurements. The detector in single-detector instruments is a photodiode, phototube, or photomultiplier tube (PMT). UV-Vis-NIR spectrometers utilize a combination of a PMT and a Peltier-cooled PbS IR detector. The light beam is redirected automatically to the appropriate detector when scanning between the visible and NIR regions. The diffraction grating and instrument parameters such as slit width can also change.
Most commercial UV-Vis absorption spectrometers use one of three overall optical designs: a fixed or scanning spectrometer with a single light beam and sample holder, a scanning spectrometer with dual light beams and dual sample holders for simultaneous measurement of P and Po, or a non-scanning spectrometer with an array detector for simultaneous measurement of multiple wavelengths. In single-beam and dual-beam spectrometers, the light from a lamp is dispersed before reaching the sample cell. In an array-detector instrument, all wavelengths pass through the sample and the dispersing element is between the sample and the array detector.
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