What is Spectrophotometry
Spectrophotometry is a technique used to measure the absorption of light by a substance as a function of its wavelength. It is commonly used in analytical chemistry and biochemistry to determine the concentration of a substance in a solution, based on the amount of light it absorbs.
The basic principle of spectrophotometry is that different substances absorb light at different wavelengths, and the amount of light absorbed is proportional to the concentration of the substance in the solution. A spectrophotometer is a device that measures the intensity of light transmitted through or absorbed by a sample as a function of wavelength.
To perform spectrophotometry, a sample is typically placed in a cuvette, which is a small, rectangular container that is optically transparent. The cuvette is then placed in the spectrophotometer, which directs a beam of light through the sample and measures the amount of light absorbed by the sample. The amount of light absorbed is then used to determine the concentration of the substance in the solution.
Spectrophotometry is a widely used technique in many fields, including biochemistry, pharmaceuticals, environmental science, and materials science, among others. It is a versatile and sensitive technique that can be used to measure a wide range of substances, including proteins, DNA, and small molecules.
What Substrates are Used for Spectrophotometry?
In general, a substrate should be optically transparent and should not interfere with the measurement of light absorption by the sample.
In biochemistry, spectrophotometry is often used to measure the concentration of proteins, DNA, and other biomolecules. In these cases, the substrate used is typically a solution containing the biomolecule of interest. For example, to measure the concentration of a protein, a solution containing the protein is placed in a cuvette and analyzed using a spectrophotometer. The amount of light absorbed by the protein is then used to determine its concentration.
In other fields, such as environmental science and materials science, spectrophotometry is used to measure the concentration of a wide range of substances, such as pollutants, metals, and dyes. In these cases, the substrate used may be a solution containing the substance of interest, or it may be a solid material that has been ground up or dissolved in a suitable solvent.
Overall, the substrate used in spectrophotometry should be selected based on the specific analysis being performed, and should be carefully prepared to ensure accurate and reproducible results.