Topic: U V Instrumentation- Source of Radiation, Wavelength Selector,
Sample Cells & Applications
CONTINUOUS ASSESSMENT 1
Presented by- Farhan Aziz
Name: Farhan Aziz
Section: B
Roll No: 15901921072
Reg No: 211590201910029
Stream: B. Pharm (4th Year, 7th Semester )
Subject: Instrumental Methods of Analysis
Subject Code: PT711
Academic Year : 2024 - 2025
COLLEGE: CALCUTTA INSTITUTE OF PHARMACEUTICAL TECHNOLOGY & A.H.S.
Vision
➤ To be a premier institute and to achieve excellence in pharmacy education and research
Mission
➤ To create state-of-the art infrastructural facilities to enhance teaching-learning and research to achieve excellence
➤ To promote innovation in teaching-learning by organizing faculty development programmes for continuous improvement in
education system to meet the needs of health-care and pharmaceutical industry
➤ To develop closer interface with pharmaceutical industry and research institutes to promote collaborative research to
ultimately become a resource centre
Programme Educational Objectives
PEO 1:- To prepare Pharmacy graduates capable of applying principles and technologies for formulation,
Development and manufacture of drugs and pharmaceuticals
PEO 2:- To produce pharmacists to serve society as the profession responsible for appropriate use of medication
and device to achieve optimal therapeutic outcomes
PEO 3:- To promote leadership and entrepreneurship qualities, human values and professional ethics.
PEO 4:- To organize workshops for hands-on-training and skill development
PEO 5:- To promote awareness on clean and green environment
❖ Contents :
Name Slide No
1. Introduction : 4
2. Instrumentation : 5
3. Source of Electromagnetic Radiation : 6-7
4. Wavelength Selectors : 8
5. Sample Cell / Cuvettes : 9
6. Detectors : 10
8. Application of UV Spectroscopy : 11
9. Conclusion : 12
9. Reference : 13
❖ Introduction :
➢ The word 'Spectroscopy' is combination of two words :
Spectroscopy
Spectrum ( Light ) Scopy ( To determine )
➢ It is the branch of science which is used for quantitative analysis of sample or matter by
using electromagnetic radiation.
➢ The instrument used for quantitative measurement is called UV- Spectrophotometer.
➢ It is used to measure the degree of absorbance of UV radiation (200-400 nm) & visible
region (400-800nm).
❖ Instrumentation :
➢ The essential components of a UV Spectrophotometer are :
1) Source of Electromagnetic Radiation
2) Wavelength Selectors / Monochromator
3) Sample Cells / Cuvettes
4) Detector
Deuterium lamp Prism
5) A Recorder & Display System Prism
Cuvettes Phototube
❖ Source of Electromagnetic Radiation :
➢ A light source should be more stable , intense & give a range of spectrum ( 180-360 nm ).
various Source of lights are :
1. Hydrogen Discharge lamp : ( 160-375 nm )
➢ A pair of electrode enclosed in glass tube, filled with
Hydrogen gas under high pressure.
High voltage current applied
Excitation of Hydrogen molecule
Electron transtion occur ground to excited state
Hydrogen Discharge lamp
Emission of UV radiation
❖ Source of Electromagnetic Radiation :
2. Deuterium Lamps :
➢ Similar as hydrogen discharge lamp. In this lamp deuterium is filled in
Place of hydrogen. It gives 3-5 times more intensity than hydrogen
Discharge lamps.
Deuterium lamp
3. Xenon Discharge Lamps :
➢ This lamp Filled with Xenon at 10-30 atmospheric pressure.
➢ It produces greater UV radiation,
4. Mercury Arc lamp : Xenon Discharge lamp
➢ In this lamp mercury vapour is stored under high pressure. The
current is applied to excited mercury atoms.
Mercury Arc Lamp
❖ Wavelength Selectors :
➢ Various components of wavelength selectors are -
Entrance Shit, Prism, Exit Shit, Filter, Diffraction grating
➢ Filter : It converts the polychromatic light into monochromatic light. It mainly 2 types :
a. Absorption Filter: Made up of glass, coated with pigments, absorb unwanted radiation.
b. Interference Filter: Composed of transparent material & thin silver film.
➢ Prism : It is more effective than filter. Generally grating monochromator prisms are used.
The prism should made of quartz.
❖ Sample Cells / Cuvettes:
➢ Sample cells or cuvettes are used to hold a sample solution.
➢ It Should be transparent , for study in UV region the cells are prepared from
Quartz. It should be filled 2 / 3 of total volume with test sample.
➢ Shape : Cylindrical or Rectangular.
➢ Path length : 1cm ( Normally ) upto 10 cm.
➢ Material : Quartz
Different shapes of cuvettes Absorption of light in cuvette
❖ Detectors:
➢ The electromagnetic radiation passed through sample cell. Some radiation are absorbed
and some amount are transmitted.
➢ This transmitted radiation fall on detector and the intensity of absorbance determined
3 types of Detectors
Photovoltaic Cell Photomultiplier tube Photo Cell
❖ Application:
➢ Various Application of Spectrophotometer are :
a. Detection of Impurities :
b. Structure Elucidation of Organic Compounds
c. Structure Analysis of Organic Compounds .
d. Effect of Conjugation can be determine.
e. Effect of Geometric Isomerism ( cis & trans ) can be determine. Spectrophotometer
f. Number of Ring in Structure can be determine.
❖ Conclusion :
➢ From this topic we can conclude that :
➢ Spectroscopy one of the most important technique for Quantitative Analysis.
➢ It is based on the absorption of radiation.
➢ The concentration of unknown sample can determined by comparing absorbance of
radiation.
❖ Reference :
➢ Dr. S. Ravi Sankar. ' UV Spectroscopy '. Text Book of Pharmaceutical Analysis. Rx
Publication, Tirunelveli(4);2010;2.1-2.10.
➢ Prof Poonam A. Salunke, Prof. Md. Rayeeb, Md. Usman et al. ' UV-Visible Spectroscopy ', A
Text book of Instrumental Method of Analysis. Pee Vee, Panjab(1);2020;19-25.
➢ Dr. S. Malathi, Dr. Pallavi Mangesh Patil, Dr. Sunil Kumar et al. ' UV-Visible Spectroscopy '.
Instrumental Method of Analysis. Thakur Publication Pvt Ltd; Lucknow(1);2019;30-35
➢ Dr. K. R. Mahadik, Dr. L. Sathiyanarayanan. ' UV-Visible Spectroscopy '. Instrumental Method
of Analysis. Nirali Prakashan, Pune(1)2020;3.1-3.8.
➢ Y. R. Sharma. ' Ultra-violet & Visible Spectroscopy '. Elementary Organic Spectroscopy.
S Chand & Company Ltd, New Delhi(5);2013;16-18.