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  1. 1. Definition A spectrophotometer is an instrument that measures the amount of light absorbed by a sample.
  2. 2. History  In Older times, it took weeks for the results to come out and, most of the time, there was only 25 percent accuracy  It was rather depressing for these scientists because quick results are very important to them, and there wasn’t any good laboratory equipment at that time to make the process faster.
  3. 3. Cont…  A scientist named Arnold J. Beckman and his colleagues at the National Technologies Laboratory (NTL) invented the Beckman DU spectrophotometer in 1940.  Results come through simple process within few minutes.  Results were 99.99 % accurate.
  4. 4. Components of Spectrophotometer
  5. 5. Light Source  to provide a sufficient of light which is suitable for marking a measurement.  Tungsten Lamp  Hydrogen Lamp  Xenon Lam
  6. 6. I) Tungsten Lamp  It is the most common light source used in spectrophotometer wavelength  range of about 330 to 900 nm  It has long life about 1200h.
  7. 7. II)Hydrogen / Deuterium Lamps For the ultraviolet region, hydrogen or deuterium lamps are frequently used. their range is approximately 200 to 450 nm. Deuterium lamps are generally more stable and has long life about 500h.This lamp generates continuous or discontinuous spectral.
  8. 8. III) Xenon flash lamps Xenon flash lamps have several advantages as the following : 1)Their range between ( 190nm - 1000 nm) 2) Emit both UV and visible wavelengths 3) Long life 4) Do not heat up the instrument 5) Reduce warm up time
  9. 9. Dispersion devices Dispersion devices causes a different wavelength of light to be dispersion at different angles. Types of Dispersion devices used are  Prism  Filters
  10. 10. Prism Prism is used to isolate different wavelength Prism may be made of glass or quartz.
  11. 11. Filters Filters separate different parts of the electromagnetic spectrum by absorbing or reflecting certain wavelengths and transmitting other wavelengths.  Absorption filters are glass substrates containing absorbing species that absorb certain wavelength. A typical example is a cut on color filter, which blocks short wavelength light, and transmits longer wavelength.
  12. 12. Absorption cells(Cuvettes) A cuvette is a kind of cell (usually a small square tube) sealed at one end, made of Plastic, glass or optical grade quartz and designed to hold samples for spectroscopic experiments.
  13. 13. Detectors Any photosensitive device can be used as a detector of radiant energy .The photocell and phototube are the simplest photodetectors, producing current proportional to the intensity of the light striking them.
  14. 14. Display devices The data from a detector are displayed by a readout device, such as an analog meter, a light beam reflected on a scale, or a digital display , Or liquid crystal display(LCD) .The output can also be transmitted to a computer.
  15. 15. Uses of Spectrophotometer  To determines the absorbance or transmission of characteristic wavelengths of radiant energy (light) by a chemical species in solution.  Identify organic compounds by determining the absorption maximum.  Used for color determination within the spectral range
  16. 16. Types of Spectrophotometer Single Beam Double Beam Split Beam
  17. 17. Single beam The single beam spectrophotometer was the first invented, and all the light passes through the sample. To measure the intensity of the incident light, the sample must be removed so all the light can pass through.  cheaper because there are less parts and the  system is less complicated.  low cost
  18. 18. Single Beam SPM
  19. 19. Double beam  A double beam spectrophotometer compares the light intensity between two light paths, one path containing a reference sample and the other the test sample.  Measurements from Double beam instruments are easier and more stable.
  20. 20. Double Beam SPM
  21. 21.  The Advantage of the double beam design Is high stability because reference and sample are measured virtually at the same moment in time.  The disadvantages are higher cost, lower sensitivity because throughput of light is poorer because of the more complex optics and lower reliability because of the greater complexity.
  22. 22. 3)Split Beam  similar to the double beam spectrophotometer but it uses a beam splitter instead of a chopper to send light along the blank  sample paths simultaneously to two separate but identical detectors
  23. 23. Split Beam SPM