The document expresses gratitude to Madam Manika Das Kotoki for the opportunity to present a presentation on spectrophotometry and for helping collect presentation data. It then provides information on spectrophotometry, including that it quantitatively measures material properties as a function of wavelength, involves the use of a spectrophotometer, and works based on Beer's Law, Lambert's Law, and Beer-Lambert's Law. It discusses the components and uses of spectrophotometers in various scientific fields and industries.

2. I express my heartful gratitude to MADAM MANIKA
DAS KOTOKI, HOD and s/t of DEPT. OF BIOTECHNOLOGY,
DARRANG COLLEGE, TEZPUR for giving me an opportunity to
present this presentation on “SPECTROPHOTOMETRY”
and helping me to collect and organize the presentation
data. I sincerely hope tat the knowledge provided by her
will be beneficial in my future life.

4.  Quantitative measurement of the reflection or
transmission properties of a material as a function of
wavelength.
 A part of BIO-INSTRUMENTATION
 Involves the use of SPECTROPHOTOMETER

5.  It is a photometer that can measure intensity as a
function of the light source wavelength.
 It is commonly used for the measurement of
transmittance or reflectance of solutions, transparent
or opaque solids, or gases.
 Also designed to measure the diffusivity on any of the
listed light ranges that usually cover around 200nm -
2500nm using different controls and calibrations.

6. A Spectrophotometer

7. Works upon the following principles-
Beer’s Law
Lambert’s Law
Beer-Lambert’s Law

8. AUGUST BEER
The intensity of a ray of Monochromatic Light passing
through an absorbing medium decreases exponentially as the
concentration of absorbing medium increases.
I = I e-klc

9. JOHANN HEINRICH LAMBERT
The intensity of light or a ray of Monochromatic Light that
passes through an absorbing medium decreases exponentially as
the length of absorbing medium increases.
I = I e-k2l

10.  The Beer–Lambert law, also known as the Lambert–Beer law or
the Beer–Lambert–Bouguer law (named after August Beer,
Johann Heinrich Lambert, and Pierre Bouguer) relates the
absorption of light to the properties of the material through
which the light is travelling.
 Two laws have been combined together as Beer – Lambert’s Law,
which is stated below-
I=I oe
-kcl

12. Wavelength
Collimator selector
(lens) (slit)
Detector
(Photo Cell)
0.20
Digital
Monochromator Sample Display
Light Source or Meter
(Prism or Grating) solution
(in Cuvette)

13. Five types of Spectrophotometer according to the range and
type of light used-
UV-Vis absorption spectrophotometer (UV-Vis)
FT-IR absorption/transmission spectrophotometer (FTIR)
Atomic absorption spectrophotometer (AAS)
Atomic fluorescence spectrophotometer (AFS)
X-ray fluorescence spectrophotometer (XFS)
Two types according to the number of beams used-
Single Beam Spectrophotometer
Double Beam Spectrophotometer

14. Manual Data Input
Computerised Data
Input

15. The use of spectrophotometers spans various scientific fields,
such as-
Physics
Materials Science
Chemistry
Biochemistry
Molecular Biology
Also used in industries including semiconductors, laser and
optical manufacturing, printing and forensic examination, as well
in laboratories for the study of chemical substances.