ULTRAVIOLET-VISIBLE

SPECTROSCOPY (UV-VIS)

SWATI V NAIR
STUDENT
DEPARTMENT OF FORENSIC SCIENCES
CTM-IRTE, FARIDABAD
 SPECTROSCOPY
Defined as the study of interaction of light with matter
 BASIC PRINCIPLE OF SPECTROSCOPY
BEER-LAMBERT’S LAW
The amount of light absorbed by the sample solution is directly proportional
to the concentration of the sample keeping the path length constant.
A∝ c
A∝ l

i.e. A= εcl
A - Absorbance I o- Intensity of incident light

ε - Molar absorption constant [M−1cm−1 ] I - Intensity of transmitted light

c - Molar concentration [M] T - Transmittance
l - Optical path length [cm generally 1cm]
 UV-VIS SPECTROSCOPY
 It is a type of Absorption Spectroscopy
 Composed of 6 essential components
1. Light sources
2. Wavelength selectors
3. Sample containers
4. Radiation detectors
5. Signal processor
6. Read out device

LIGHT WAVELENGTH RADIATION READ OU

SAMPLE CONTAINERS SIGNAL PROCESSOR DEVIC
OURCE SELECTORS DETECTORS
1. LIGHT SOURCE
A. Tungsten Filament- VIS B. Hydrogen-Deuterium lamp - UV
2. Wavelength Selector
A. PRISM B. DIFFRACTION GRATING

C.FILTER
3. Sample Compartment
Cells with an internal diameter of 1 cm.
• UV - QUARTZ
• VISIBLE - GLASS/ PLASTIC/ GLASTIC
4. Detectors
A. PHOTO-EMISSIVE VACUUM TUBE B. PHOTODIODES - Solid-State Devices
[Semiconductors]
[Least preferred detector for UV-VIS spectroscopy]
C. PHOTO MULTIPLIER TUBE
• Most used detector for this spectroscopy
• Contains a series of dynodes connected to a very high voltage system
• Contains a photo emissive cathode and an anode
WORKING OF SPECTROPHOTOMETER
 GRAPHS/OUTPUT

A. QUALITATIVE ANALYSIS B. QUANTITATIVE ANALYSIS

 APPLICATIONS
1. GENERAL
• Measuring the absorbance of hemoglobin to determine hemoglobin concentrations may be
used in cancer research.
• In wastewater treatments, UV-Vis spectroscopy can be used in kinetic and monitoring
studies to ensure certain dyes or dye by‑products have been removed properly by comparing
their spectra over time.
• It also finds great utility in food authenticity analysis and air quality monitoring.
• Tracking changes in the wavelength corresponding to the peak absorbance is useful in
examining specific structural protein
• Shifts in peak absorbance wavelengths can also be useful in more modern applications such
as characterization of very small nanoparticles.
 APPLICATIONS
2) FORENSIC
• DNA And RNA Analysis
• Pharmaceuticals Analysis
• Beverage Analysis
• Used As Detectors In Gc,hplc
 REFERENCES
1. Harris DC. Quantitative Chemical Analysis. 7th ed, 3rd printing. W. H. Freeman; 2007.

2. Diffey BL. Sources and measurement of ultraviolet radiation. Methods. 2002;28(1):4-13. doi:10.1016/S1046-2023(02)00204-9

3. Namioka T. Diffraction Gratings. In: Vacuum Ultraviolet Spectroscopy. Vol 1. Experimental Methods in Physical Sciences. Elsevier; 2000:347-377.
doi:10.1016/B978-012617560-8/50018-9

4. Mortimer Abramowitz and Michael W. Davidson. Photomultiplier Tubes. Molecular Expressions. Accessed April 25, 2021.
https://micro.magnet.fsu.edu/primer/digitalimaging/concepts/photomultipliers.html

5. Picollo M, Aceto M, Vitorino T. UV-Vis spectroscopy. Phys Sci Rev. 2019;4(4). doi:10.1515/psr-2018-0008

6. https://byjus.com/chemistry/uv-vis-spectroscopy/

7. Cole, Kenneth; Levine, Barry S. (2020), Levine, Barry S.; KERRIGAN, SARAH (eds.), "Ultraviolet-Visible Spectrophotometry", Principles of
Forensic Toxicology, Cham: Springer International Publishing, pp. 127–134, doi:10.1007/978-3-030-42917-1_10, ISBN 978-3-030-42917-1,
retrieved 19 October 202

8. Jump up to:a b Vitha, Mark F. (2018). "Chapter 2". Spectroscopy: principles and instrumentation. Hoboken, NJ: John Wiley & Sons. ISBN
978-1-119-43664-5.

9. Edwards, Alison A.; Alexander, Bruce D. (1 January 2017), Lindon, John C.; Tranter, George E.; Koppenaal, David W. (eds.),
"UV-Visible Absorption Spectroscopy, Organic Applications", Encyclopedia of Spectroscopy and Spectrometry (Third Edition), Oxford: Academic Press,
pp. 511–519, doi:10.1016/b978-0-12-803224-4.00013-3, ISBN 978-0-12-803224-4, retrieved 19 October 2023
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