Spectrophotometry: A Comprehensive Guide
Discover the principles and components of spectrophotometry, including the interaction between light and samples, the role of spectrophotometers, and the use of different light sources and sample cells. Learn about the applications of spectrophotometry in both qualitative and quantitative analysis, as well as the importance of sample and standard solutions in concentration measurement.
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Presentation Transcript
Principle : there is interaction between the light and sample particle , spectrophotometer is employed to measure the amount of light that a sample absorbs .
spectrophotometer consists of two instruments: 1- spectrometer for producing light of any selected color (wavelength 2- photometer for measuring the intensity of light. The instruments are arranged so that liquid in a cuvette can be placed between the spectrometer beam and the photometer
The most common spectrophotometers are used in the UV and visible regions of the spectrum .
Light source : Two kinds of lamps 1- Deuterium lamp in UV- spectrophotometer 2- tungsten lamp in visible - spectrophotometer
Sample cell : A container that contains a sample is usually called "cell two types are available: 1- glass used in visible spectrophotometer 2- quartz used in UV spectrophotometer .
Application : 1- qualitative : The wavelength of maximum conc. give information about structure in max 2- quantitative : Determination of conc. By beer s law A = abc
Sample and standard solution : To measure the concentration of a substance, a sample of (unknown concentration) is compared with known concentration (standard solution) to determine the concentration . The blank contain all the substance except the analyte
