Ultraviolet–visible spectroscopy
Ultraviolet–visible spectroscopy (UV-Vis spectroscopy) is a type of absorption spectroscopy in the ultraviolet and visible regions of the electromagnetic spectrum. This technique is used to measure the absorbance of light by a sample, which can provide information about the sample's chemical composition and concentration.
Principle[edit | edit source]
UV-Vis spectroscopy is based on the principle that molecules absorb light at specific wavelengths. When a molecule absorbs UV or visible light, electrons in the molecule are excited from a lower energy level to a higher energy level. The amount of light absorbed at each wavelength is measured and plotted to create an absorption spectrum.
Instrumentation[edit | edit source]
The main components of a UV-Vis spectrophotometer include:
- A light source that emits light in the UV and visible regions.
- A monochromator to select specific wavelengths of light.
- A sample holder where the sample is placed.
- A detector to measure the intensity of transmitted light.
Applications[edit | edit source]
UV-Vis spectroscopy is widely used in various fields, including:
- Chemistry: To determine the concentration of a substance in a solution using Beer-Lambert law.
- Biochemistry: To study the properties of proteins, nucleic acids, and other biomolecules.
- Environmental science: To analyze pollutants in water and air.
- Pharmaceutical industry: To ensure the quality and consistency of drugs.
Beer-Lambert Law[edit | edit source]
The Beer-Lambert law relates the absorbance of light to the properties of the material through which the light is traveling. The law is expressed as: \[ A = \varepsilon \cdot c \cdot l \] where:
- \( A \) is the absorbance,
- \( \varepsilon \) is the molar absorptivity,
- \( c \) is the concentration of the solution,
- \( l \) is the path length of the sample cell.
Advantages and Limitations[edit | edit source]
Advantages[edit | edit source]
- Non-destructive analysis.
- High sensitivity and specificity.
- Rapid and simple to perform.
Limitations[edit | edit source]
- Requires clear and colorless samples.
- Limited to compounds that absorb UV or visible light.
- Interference from other absorbing species in the sample.
See also[edit | edit source]
References[edit | edit source]
External links[edit | edit source]
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Contributors: Prab R. Tumpati, MD