University Institute of Medical Lab
Technology (UIMLT)
Course Name: MEDICAL LAB
INSTRUMENTATION
Course Code: MLTMI-901|11
Teacher’s Name :Ali Ahsan
Designation: Lecturer
THE UNIVERSITY OF LAHORE
FLAME EMISSION
SPECTROPHOTOMETER QUALITY
CONTROL
Learning Objectives:
• Definition
• Monochromator
• Determination of unknown concentration
using calibration graph
• Sources of errors
• Quality control
• Applications
• Limitation
FLAME EMISSION
SPECTROPHOTOMETER
• The number of atoms of an element
excited by the flame depends on :
– Flame temperature
– The energy difference between the
excited and ground state
– Any increase of the flame temperature
is accompanied by great increase in the
number of excited atoms
MONOCHROMATOR
• PURPOSE
– To select a particular wavelength
– And absorbs all the other wavelengths
TYPES OF MONOCHROMATOR
– Prisms
– Interference filters
– Diffraction grating
DETECTORS
• Purpose
– Convert radiant transmitted energy into
an electrical signal
• Types
– Photocell
– Photo-emission tubes
– Photomultiplier tubes
METER
It displays the intensity of the emitted
light(higher the concentration →higher the
intensity of emitted light→higher the meter
reading
Output Devices
• Galvanometer
• Digital display
• Recorder
A BRIEF OVERVIEW
• The solvent is first evaporated leaving
fine divided solid particles
• This solid particles move towards the
flame, where gaseous atoms and ions are
produced
• The ions absorbs the energy from the
flame and excited to high energy levels,
A BRIEF OVERVIEW
• When the atoms return to the ground state
radiation of the characteristics element is
emitted
• The intensity of emitted light is related to
the concentration of the element
• The temperature of the flame must b
2000°K or above. The fraction of the
number of atoms which are excited is
given below by Boltzman equation:
DETERMINATION OF UNKNOWN
CONCENTRATION USING
CALIBRATION GRAPH
• The calibration graph is plotted with the
concentration against intensity of emitted
light(meter reading).
• The sample of unknown concentration(Cu) is
sprayed over the flame and the intensity of
emitted light(Eu)(meter reading) is noted.
• From the calibration graph, the unknown
concentration (Cu) can be found.
DETERMINATION OF THE UNKNOWN
CONCENTRATION OF THE GIVEN
SAMPLE(NA+)
• The wavelength is fixed at 589nm using the
monochromator.
• The blank sample is sprayed over the flame
and the meter reading is set to zero.
• Now the series of standard samples of known
concentrations are sprayed one by one and the
meter readings are noted.
Concentration Meter reading
Blank(Co) 0
C1 E1
C2 E2
C3 E3
C4 E4
C5 E5
SOURCES OF ERRORS
• Flame Temp:
– High : Ionization
– Low : insufficient atoms will be excited
– Aspiration Rate:
• Protein coating on atomizer walls
• Highly concentrated samples
QUALITY CONTROL
• Changes in temp.
• Changes in external light near the
instrument
• Accumulation of light-emitting material
on burner
• These effects can be monitored by
sampling water at intervals
– CHANGE IN SENSITIVITY
•Change in flow rate through capillary
•Variation in flame temp.
•Alterations in sample chamber such
as accumulation of solid matter on the
walls
• These effects are monitored by sampling
the standard after zero checking
APPLICATIONS
• Flame photometry is specially used in
accurate analysis of alkali and alkaline
earth metals.
• The process of analysis is very simple.
• The flame photometry is used in the
quantitative analysis by using standard
Curve method.
• The amount of metals present in the waste
water (especially alkali and alkaline earth
metals ) can be detected.
• With the help of flame photometry we can
determine the hardness of water.
• It is used to determine Na, K, Ca, Zn present
in the cement
• It is used in the determination of lead present
in the petrol
• The phenomenon is used in the determination
of tetra ethyl lead(TEL)and Manganese
present in the gasoline stock.
• We can determine the amount of baron present
in biological fluids and tissues can be done
with the help of flame photometry.
LIMITATIONS
• It gives the total metal content present in the
soln it does not give any idea about the
molecular conditions of metals.
• It is used for the analysis of alkali and
alkaline earth metal ,specially.
• Non-radiating elements like carbon ,
hydrogen and halogens cannot be detected
by this method.
• Only solutions can be analyzed.
• For alkali and alkaline earth metals ,low
temperature of the flame is required,
because their I.P is low so at higher
temperature those metals are ionized.
• This ionization decreases the intensity of
light emitted, so low temperature is
required.
FLAME EMISSION
SPECTROPHOTOMETER VS
ATOMIC ABSORPTION
SPECTROPHOTOMETER
LEARNING OUTCOMES:
After completion of this lecture graduates will
be able to:
• Know the Definition
• Understand the Monochromator
• Determination of unknown concentration
using calibration graph
• Understand the Sources of errors, Quality
control, Applications and Limitations of
Flame EmissionSpectrophotometer
REFERENCES
•Fody EP., Bishop ML. Clinical
Chemistry: Principles, Techniques, and
Correlations. 7th ed. 2013.
•Anwar M. Manual of laboratory
medicine. 4th ed. Armed Forces Institute
of Pathology Rawalpindi-Pakistan; 2005
•Prudent Practices in the Laboratory,
Handling and Management of Chemical
Hazards. National Academies Press
(US); 2011.
Thank You
ANY QUESTION???
