SPECTROPHOTOMETRY. Principle : there is interaction between the light and sample particle, spectrophotometer is employed to measure the amount of light.

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Presentation transcript:

SPECTROPHOTOMETRY

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.UVvisible

Light source : Two kinds of lamps 1- Deuterium lamp in UV- spectrophotometer spectrophotometer - 2- tungsten lamp in visible

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 λ 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