9/29/21, 8:35 PM Spectrophotometer- Principle, Instrumentation, Applications

Menu Search Notes

Home » Instrumentation » Spectrophotometer- Principle, Instrumentation, Applications

Spectrophotometer- Principle, Instrumentation, Applications

July 8, 2021 by Sagar Aryal

Table of Contents 
What is a spectrophotometer?
Principle of Spectrophotometer
Instrumentation of Spectrophotometer
Applications
References
Spectrophotometer- Principle, Instrumentation, Applications

Park Systems
OPEN
Park FX40 | The Autonomous AFM

Ida Slams the Northeast with Tornadoes, Torr…

What is a spectrophotometer?
A spectrophotometer is an instrument that measures the amount of light absorbed by a
sample.
Spectrophotometer techniques are mostly used to measure the concentration of solutes in
solution by measuring the amount of the light that is absorbed by the solution in a cuvette
placed in the spectrophotometer.
Scientist Arnold J. Beckman and his colleagues at the National Technologies Laboratory
(NTL) invented the Beckman DU spectrophotometer in 1940.

https://microbenotes.com/spectrophotometer-principle-instrumentation-applications/ 1/12
9/29/21, 8:35 PM Spectrophotometer- Principle, Instrumentation, Applications

FEATURED BIOLOGY VIDEOS

Ida Slams the Northeast with Tornadoes, Torr…

Principle of Spectrophotometer
The spectrophotometer technique is to measure light intensity as a function of wavelength. It
does this by diffracting the light beam into a spectrum of wavelengths, detecting the intensities
https://microbenotes.com/spectrophotometer-principle-instrumentation-applications/ 2/12
9/29/21, 8:35 PM Spectrophotometer- Principle, Instrumentation, Applications

with a charge-coupled device, and displaying the results as a graph on the detector and then on
the display device.

1. In the spectrophotometer, a prism (or) grating is used to split the incident beam into
different wavelengths.
2. By suitable mechanisms, waves of specific wavelengths can be manipulated to fall on the
test solution. The range of the wavelengths of the incident light can be as low as 1 to 2nm.
3. The spectrophotometer is useful for measuring the absorption spectrum of a compound,
that is, the absorption of light by a solution at each wavelength.

Park FX40 | The

Autonomous AFM
Park Systems

Instrumentation of Spectrophotometer
The essential components of spectrophotometer instrumentation include:

1. A table and cheap radiant energy source

Ida Slams the Northeast with Tornadoes, Torr…

Materials that can be excited to high energy states by a high voltage electric discharge (or)
by electrical heating serve as excellent radiant energy sources.

2. A monochromator, to break the polychromatic radiation into component wavelength (or)

bands of wavelengths.

A monochromator resolves polychromatic radiation into its individual wavelengths and

isolates these wavelengths into very narrow bands.

Prisms:

https://microbenotes.com/spectrophotometer-principle-instrumentation-applications/ 3/12
9/29/21, 8:35 PM Spectrophotometer- Principle, Instrumentation, Applications

A prism disperses polychromatic light from the source into its constituent wavelengths by
virtue of its ability to reflect different wavelengths to a different extent
Two types of Prisms are usually employed in commercial instruments. Namely, 600 cornu
quartz prism and 300 Littrow Prism.

Grating:
Gratings are often used in the monochromators of spectrophotometers operating
ultraviolet, visible and infrared regions.

3. Transport vessels (cuvettes), to hold the sample

Samples to be studied in the ultraviolet (or) visible region are usually glasses (or) solutions
and are put in cells known as “CUVETTES”.
Cuvettes meant for the visible region are made up of either ordinary glass (or) sometimes
Quartz.

4. A Photosensitive detector and an associated readout system

Most detectors depend on the photoelectric effect. The current is then proportional to the
light intensity and therefore a measure of it.
Radiation detectors generate electronic signals which are proportional to the transmitter
light.
These signals need to be translated into a form that is easy to interpret.
This is accomplished by using amplifiers, Ammeters, Potentiometers and Potentiometric
recorders. Ida Slams the Northeast with Tornadoes, Torr…

https://microbenotes.com/spectrophotometer-principle-instrumentation-applications/ 4/12
9/29/21, 8:35 PM Spectrophotometer- Principle, Instrumentation, Applications

Park Systems
OPEN
Park FX40 | The Autonomous AFM

Applications
Some of the major applications of spectrophotometers include the following:

Detection of concentration of substances

Detection of impurities
Structure elucidation of organic compounds
Monitoring dissolved oxygen content in freshwater and marine ecosystems
Characterization of proteins
Detection of functional groups
Respiratory gas analysis in hospitals
Molecular weight determination of compounds
The visible and UV spectrophotometer may be used to identify classes of compounds in
both the pure state and in biological preparations.

Ida Slams the Northeast with Tornadoes, Torr…

Ad

Park FX40 | The Autonomous AFM

Park Systems OPEN

https://microbenotes.com/spectrophotometer-principle-instrumentation-applications/ 5/12
9/29/21, 8:35 PM Spectrophotometer- Principle, Instrumentation, Applications

References
1. https://www.biochemden.com/spectrophotometer-instrumentation-principle/
2. https://www.azom.com/article.aspx?ArticleID=10245
3. https://web.stanford.edu/class/chem184/lectures08/Zare_Spectroscopy.pdf
4. https://byjus.com/chemistry/spectrophotometer-principle/
5. https://www.slideshare.net/rey216/spect

Ad

Park FX40 | The Autonomous AFM

Park Systems OPEN

Spectrophotometer- Principle, Instrumentation, Applications

Read Also ...

UV Spectroscopy- Principle, Instrumentation, Applications
Electron Spin Resonance (ESR)- Principle, Instrumentation, Applications
Mass Spectrometry (MS)- Principle, Working, Instrumentation, Steps,
Ida Slams the Northeast with Applications
Tornadoes, Torr…

X-Ray Spectroscopy- Principle, Instrumentation and Applications

Chromatography- definition, principle, types, applications
Simple Microscope- Definition, Principle, Parts, Applications
Centrifugation- Principle, Types and Applications
Simple diffusion- definition, principle, examples, applications
Romanowsky Stains- Principle, Types, Applications
Silver Staining- Principle, Procedure, Applications

https://microbenotes.com/spectrophotometer-principle-instrumentation-applications/ 6/12
9/29/21, 8:35 PM Spectrophotometer- Principle, Instrumentation, Applications

Park Systems
OPEN
Park FX40 | The Autonomous AFM

Instrumentation
Spectrophotometer, Spectrophotometer Applications, Spectrophotometer Components,
spectrophotometer diagram, spectrophotometer function, spectrophotometer how to use,
Spectrophotometer Instrumentation, spectrophotometer lab, spectrophotometer lab procedure,
Spectrophotometer Parts, Spectrophotometer Principle
Phylum Platyhelminthes- characteristics, classification, examples
Phylum Arthropoda- Characteristics, classification, examples

10 thoughts on “Spectrophotometer- Principle, Instrumentation, Applications”

Saad Zafar
August 23, 2021 at 10:49 AM
Ida Slams the Northeast with Tornadoes, Torr…

What is the difference b/w Sample solution and reference blank? why we use reference
standard in in Spectrophotometer??

Reply

Olusegun teniola
March 26, 2021 at 4:27 PM

https://microbenotes.com/spectrophotometer-principle-instrumentation-applications/ 7/12
9/29/21, 8:35 PM Spectrophotometer- Principle, Instrumentation, Applications

It really help alot

Reply

Dilini nisansala
October 24, 2020 at 12:12 PM

thank you for giving this more information about specrophotometer. Actually, It very
helps for answering questions me.

Reply

Manzoor Ullah
October 20, 2020 at 10:16 AM

Interesting

Reply

Ida Slams the Northeast with Tornadoes, Torr…

Tonia Tonia
February 14, 2020 at 6:14 AM

Nice one

Reply

https://microbenotes.com/spectrophotometer-principle-instrumentation-applications/ 8/12
9/29/21, 8:35 PM Spectrophotometer- Principle, Instrumentation, Applications

Jahan Sher
July 11, 2020 at 11:29 PM

Spectrophotomer or Spectrophotometery ya dono same haiin kia

Reply

Kajal
September 1, 2020 at 10:44 AM

Spectrophotometer bis the device

Spectrophotometery is the technique

Reply

Arpita
September 8, 2019 at 3:48 PM

Ida Slams the Northeast with Tornadoes, Torr…

Thanks fr giving the sufficient knowledges about Spectrophotometer….

Reply

munmun
May 4, 2019 at 9:57 AM

https://microbenotes.com/spectrophotometer-principle-instrumentation-applications/ 9/12
9/29/21, 8:35 PM Spectrophotometer- Principle, Instrumentation, Applications

Please be more scientific, I guess this is not a complete explanation of principle of

spectrophotometer.

Reply

Ali Chenak
October 16, 2018 at 6:01 PM

Thanks

Reply

Leave a Comment

Ida Slams the Northeast with Tornadoes, Torr…

Name *

Email *

Save my name, email, and website in this browser for the next time I comment.

Post Comment

This site uses Akismet to reduce spam. Learn how your comment data is processed.
https://microbenotes.com/spectrophotometer-principle-instrumentation-applications/ 10/12
9/29/21, 8:35 PM Spectrophotometer- Principle, Instrumentation, Applications

Topics / Categories

Select Category

Read Also (Related Notes)

UV Spectroscopy- Principle, Instrumentation, Applications

Electron Spin Resonance (ESR)- Principle, Instrumentation, Applications
Mass Spectrometry (MS)- Principle, Working, Instrumentation, Steps, Applications
X-Ray Spectroscopy- Principle, Instrumentation and Applications
Chromatography- definition, principle, types,Ida Slams the Northeast with Tornadoes, Torr…
applications

Advertise with us
Our Team
About Us
Contact Us

Disclaimer (Terms and Conditions)

Privacy Policy

https://microbenotes.com/spectrophotometer-principle-instrumentation-applications/ 11/12
9/29/21, 8:35 PM Spectrophotometer- Principle, Instrumentation, Applications

Cookie Policy

© 2021 Microbe Notes. Made with ♡ by Sagar Aryal.

Ida Slams the Northeast with Tornadoes, Torr…

https://microbenotes.com/spectrophotometer-principle-instrumentation-applications/ 12/12