34 Flame Photometry

Course Code : PDA302T
Pharmaceutical Analysis
Session
Flame Photometry
1
aculty of Engineering
Faculty of Pharmacy& Technology M. S. Ramaiah University of Applied Sciences
Session Objectives
By the end of this session, students will be able to:
2
Atomic Spectroscopy
3
Atomic Spectroscopy Methods
1. Atomic Emission Spectroscopy (Flame Photometry)

2. Atomic Absorption Spectrometry (AAS)
4
Atomic Spectroscopy
To understand the relationship of these techniques to each other, It is important to
understand the atom itself and the atomic process involved in each technique.
Ground state atom (stable or

Excited
normal orbitalstate atom
configuration)
3
Excited Excited
states states 3
Spectral resonance line (The

strongest line)
3
Ground state Ground state 5
Faculty of Pharmacy& Technology
Light energy
M. S. Ramaiah University of Applied Sciences
Atomic Spectroscopy
Practically, the ratio of the excited to ground state atoms is extremely small. Therefore, The absorption
spectrum is usually only associated with transitions from the ground state to higher energy states
~0% population Excited state atoms

Energy
Emission Absorption
~100% population Ground state atoms
Boltzmann equation explain the relatioship between the ground and excited state atoms N1/N =
(g1/g)e-E/KT
N1: No. of excited atoms N: No. of ground state atoms E: excitation energy
6
Atomic Spectroscopy
The process of excitation and decay to ground state is involved
in the two techniques of atomic spectroscopy.
We measure the energy absorbed or emitted and use it for
quantification process
7
Atomic Emission Spectroscopy (AES)
(Flame Photometry)
Principle: Flame photometry is based upon those particles that are electronically
excited in the medium.
Flame : is the source of excitation energy. (low energy source).
Uses:
Flame photometry is used mainly for the determination of alkali metals and easily
excited elements (Na, K, Li, Ca, etc.) particularly in biological fluids and tissues
8
Flame Photometry
Events occuring in FES
Solution of metal Ground state atoms

salt (NaCl) Na
Solvent evaporation Emission

and return
Solid aerosol of to G
Excited atoms
metal salt (NaCl) Na*
Volatilization or Excitation
decomposition
Gaseous metal Atomization
salt (NaCl) Free atoms
Na
9
Flame Photometry
Instrument
components Monochromator
Flame Detector
Flame Structure Secondary combustion

zone
Interzonal region
Primary combustion
zone
Fuel- oxidant mixture

10
Flame Photometry
Functions of Flame
1. To convert the constituents of liquid sample into the vapor state.
2. To decompose the constituents into atoms or simple molecules:
M+ + e- (from flame) M + h
3. To electronically excite a fraction of the resulting atomic or molecular species
M M* Oxidant - Fuel Max. temp. (oC)
Air- propane 1725
Air- acetylene 2400
Oxygen- acetylene 3100
The flame is composed of :
Nitrous oxide-acetylene 3000
a fuel gas and oxidant gas
Air-hydrogen 2000
Oxygen-hydrogen 2700
Air + argon -hydrogen 1577 11
Flame Photometry
Factors affecting intensity of flame emission :
1- The concentration of the analyte in solution
2- The rate at which excited atoms are formed in the flame.
3- The rate at which the sample is introduced into the flame.
4- Temperature of the flame.
5- Composition of the flame.
6- The ratio of fuel to oxidant in the flame.
7- Solvent used to dissolve the sample.
The flame temperature is the most important factor. Increase in flame temperature causes
an increase in emission intensity. This is controlled by composition of the flame.
High temperature flames should not be used for elements that ionized easily e.g. Na, K, Li
or Ce. However, high temperature flames are generally favored for transition elements and
alkaline earth metals. 12
Flame Photometry
Effect of the solvent used to dissolve the sample ; if the solvent is water the process is
slow and if it is organic solvent the process is fast and emission intensity is increased.
It is therefore very important that calibration curves be prepared using the same
solvent.
The stochiometric ratio of fuel to oxidant in the flame must be used, in which both
fuel to oxidant are totally consumed.
13
Flame Photometry
The nebulizer-burner system
To convert the test sample into gaseous atoms
Nebulizer produce an aerosol of the test solution
Burner in which the mixing between fuel and oxidant
Types of burner system

Flame
1. Pre-mix or laminar flow burner Fuel in
Burner
Advantages
1. Homogenous flame Oxidant in
2. Suitable for AAS and AES as the pathway
could be increased
Disadvantages Sample
Suffers from explosion hazards Aspirator air Mixing

14 baffles
Drain
Flame Photometry
2. Total consumption burner
3 concentric tubes, the sample, fuel and oxidant only
mix at the tip of burner
Used mainly for FES (short bath)
Fuel
Oxidant
Advantages
1. Simple to manufacture
Sample
2. Allows a total representative sample to reach the
flame
3. Free from explosion hazards Oxidant
Fuel
Disadvantages
1. Aspiration rate varies with different solvents
2. Suitable only for AES
15
Flame Photometry
Non Flame Atomizers
For example: Heated Granite Furnace
Sample evaporation time and temp. controlled drying and ashing
Advantages
1. small samples are analysed
2. 1000-fold more sensitive than flame
3. Oven is adaptable to determination of solid samples
Disadvantages
1. Low accuracy 2. Low precision

2. More ionic interferences due to very high temp.
16
Flame Photometry
Monochromators As in UV
Detectors Films or photomultipliers
Analytical technique
1. Choice of the wavelength: of max. sensitivity and min. spectral interferences
2. Sample preparation:
a. It is very important to obtain the sample in a form of solution, where the spectral and chemical interferences
are absent
b. Demineralized dist. Water and very pure reagents are to be used because of the high sensitivity of the
technique
c. Because of the instability of the very dil. Solution, it is advisable to dilute the soln just before use.
d. Several elements can be determined in blood, urine, cerebrospinal fluid and other biological fluids by direct
aspiration of the sample after dilution with water.
17
Flame Photometry
Chemical interferences: can often be overcome by simple dilution with a suitable
reagent solution e.g. serum is diluted by EDTA solution for the determination of
calcium in order to prevent interference from phosphate.
3. Standard curves
b s o rb a n c e ,
Deviations from linearity
may occur AIntensity
A
Faculty of Pharmacy& Technology
Concentration, c 18
M. S. Ramaiah University of Applied Sciences
Flame Photometry
3. Qualitative analysis
Flame photometry are useful mostly for the detection of elements in group I and II of the
periodic table. The presence of certain elements can be detected by the use of a filter or
monochromator.
Advantages and disadvantages
The method is not as reliable as other atomic emission spectroscopic methods, but it is fast
and simple.
4. Quantitative analysis
To perform quantitative analysis, the sample is introduced into the flame and the intensity of
radiation is measured. The concentration of the emitting substance is then calculated from a
calibration curve or using standard addition method.
19
Flame Photometry
Application of flame photometry in pharmaceutical analysis
1. Metals are major constituents of several pharmaceuticals such as dialysis solutions, lithium carbonate
tablets, antacids and multivitamin - mineral tablets.
2. The elements Na, K, Li, Mg, Ca, Al and Zn are among the most common elements subjected to
pharmaceutical analysis using flame emission technique.
3. Sodium and potassium levels in biological fluids are difficult to analyze by titrimetric or colorimetric
techniques. Their analysis is very important for control of infusion and dialysis solutions which must be
carefully monitored to maintain proper electrolyte balance.
Advantages and disadvantages

1. Flame emission is the simplest and least expensive technique.
2. The analysis may be carried out without prior separation as other components such as dextrose, do not
interfere.
20
Atomic Absorption Spectroscopy
Atomic Absorption spectroscopy involves the study of the absorption of radiant energy by
neutral (ground state) atoms in the gaseous state.
Instrument components
Hollow cathode P0 P
Monochromator
lamp
Detector
Sample
Source
Lamps low-pressure inert gas
Hollow Cathod lamp Inert carrier gas

Ne or Ar
21
22
Electrodless Discharge Lamps, EDL
For easily evaporized elements as Hg or As
Used for AAS and AES
Give much greater radiation intensities than hollow cathode
There is no electrode, but instead , the inert carrier gas is energized by an intense field
of radiofrequency or microwave radiation plasma formation which cause excitation
of the metal inside
23
Degree of absorption:
Total amount of light absorbed = (e2/mc2)Nf

Where:
e = electronic charge, m = mass of electron
c = speed of light, N = total No. of atoms that can absorb light
f = Ability of each atom to absorb light
, e, m, and c are constants, therefore
Total amount of light absorbed = constant x Nf

Since f is also constant for the same substance
A& C
24
25
Interferences
Spectral Interferences
1. They arise when the absorption line of an interfering species either overlaps or lies so
close to the analyte absorption line that resolution by the monochromator becomes
impossible. Ex. Mg in presence of Ca.
2. They occur from band or continuous spectra which are due to absorption of molecules or
complex ions remaining in the flame
3. They arise from flame background spectrum.
Correction:
1. It may be useful to shift to another spectral line
2. Two line correction method: (Instrumental correction)
It employs a line from the source as a reference. The line should lie as close as possible to the analyte line
but must not be absorbed by the analyte. If the conditions are met, any decrease in the reference line
from that observed during calibration arises from absorption by the matrix of the sample. 26
Chemical Interferences
occurs during atomization that prevent the gaseous atoms production of the analyte.
They are more common than spectral ones.
Types of chemical interferences

1. Formation of stable compounds: incomplete dissociation of the sample in flame
2. Formation of refractory oxides: which fail to dissociate into the constituent atoms
Examples 1. Detn. of Ca in presence of sulphate or phosphate
2. Formation of stable refractory oxides of TiO2, V2O5 or Al2O3 by reaction with O2 and
OH species in the flame
Overcome
1. Increase in the flame temp. Formation of free gaseous atoms
e.g. Al2O3 is readily dissociated in acetylene-nitrous oxide flame 27
2. Use of releasing agents: M-X + R RX + M ex. Detn of Ca in presence of
phosphate
(Ca - phosphate + SrCl2 Sr-phosphate + Ca atoms) or (Ca phosphate +
EDTA Ca-EDTA easily dissociated complex ).
3. Solvent extraction of the sample or of the interferring elements

Ionization Interferences
Ionization of atoms in the flame decrease the absorption or emission
Overcome : 1. Use of lowest possible temp which is satisfactory for the sample ex.
Acetylene air must not be used for easily ionised elements as Na, K, Ca, Ba
2. Addition of an ionisation supressant ( soln of cation has a lower ionisation potential
than that of the sample, e.g. addition of K-soln to Ca or Ba soln. Ca Ca2+ + 2e K
K+ + e
28
Physical Interferences
1. Variation in gas flow rate
2. Variation in sample viscosity
3. Change in flame temp.
Overcome: 1. by continuous calibration

2. Use of internal standard
Advantages of AAS: Very sensitive.

Fast.
Disadvantages of AAS: Hollow cathode lamp for each element.

Expensive element. 29
Relationship Between Atomic Absorption and Flame Emission
Spectroscopy
Atomic Absorption Flame Emission
1. Measures the radiation absorbed by 1. Measures the radiation emitted by the
the unexcited atoms excited atoms
2. Depends only on the number of 2. Depends only on the number of

unexcited atoms excited atoms
3. Absorption intensity is NOT affected 3. Emission intensity is greatly affected

by the temperature of the flame by the temperature variation of the flame
30
Atomic Emission Spectroscopy
Using Non-Flame excitation sources
1. There is no single excitation source can excite all elements
2. The emitted radiation usually consists of sharp well defined lines, which fall
in UV or visible region
3. Identification of the of these lines permits qualitative analysis of these
elements, whereas measurements of their intensities permits quantitative
analysis
Advantages
1. Excellent method for trace element analysis at ppm level

2. Used nearly for all elements in periodic table
31
3. Used for very small samples, even less than 1 mg
4. There is no need for prior separation
5. Relatively rapid technique
Disadvantages 1. Expensive
2. Low precision and accuracy
3. Destroying the sample
4. Used mainly for metals
High energy excitation sources

Plasma excitation sources
Laser
Arc and spark emission spectrometry (Spectrography)
Microwave and x-ray 32
Plasma excitation sources
1. A plasma is a cloud of highly ionized gas containing significant numbers of positive and negative ions,
free electrons and neutral particles.
2. Plasma sources operate at high temperatures between 7000 and 15000 K. Thus, it produces a greater
number of excited emitted atoms, especially in the UV region, than that produced by flame.
3. Using this technique, excitation operates through a plasma produced electrically in a carrier gas such as
nitrogen or argon.
3. The main types of argon plasma sources

a. Inductively coupled plasma; ICP
b. Direct current plasma; DCP
c. A microwave-induced plasmas is recently introduced to spectro-chemical analysis methods.
33
Plasma
Induction coils Magnetic field
Quartz tubes
Argon tangential flow
Sample flow 34
Inductively coupled plasma; ICP
Argon gas flows upward through a quartz tube, around which is wrapped with a copper
or selenoid induction coil.
The coil is energized by a radio frequency AC generator creating a changeable magnetic

field on the flowing gas inside. This induces a circulating current in the gas, which in
turn heats it.
Argon is not a conductor at low temperatures, but becomes electrically conducting by

heating it. The induction is initiated by arc or a heated graphite rod.
It is used for multi-element determination
35
Direct current, DCP
It consists of a high-voltage discharge between two graphite electrodes. The recent design
employs a third electrode arranged in an inverted Y-shaped which improves the stability
of discharge.
The sample is nebulised at a flow rate of 1 ml/min. Argon is used as carrier gas. The
argon ionized by the high-voltage is able to sustain a current.
DCP generally has lower detection limits than ICP. However, DCP is less expensive than
ICP.
36
Advantages of plasma excitation source:
1- The sample could be introduced in solution form through a nebulizer (easy for
quantitative analysis).
2- It is suitable for quantitative multielement determinations
3- The high temperature of plasma eleminates many chemical interferences present

in a flame
4- It is well suited for refractory (oxide forming) elements e.g. P, Ur and tungeston
and for difficult-to-excite elements such as Zn and Cd.
5- The emission intensity-versus-cencentration range is linear over a very wide

dynamic ranges of analytes.
37
Laser excitation source
Laser beam is used to vaporize the sample, which is then excited electrically.
The sample is loaded just beneath the two electrodes that will be used to generate the
electrical discharge.
A ruby laser is then focused through a microscope onto the surface. The energy from the
laser causes an intense local hot spot which vaporizes a small quantity of sample.
The vapor circuits the electrodes and electrical discharge occurs which excites the metals
in vapor. The excited metals emit typical emission spectra which are collected and
measured as usual.
38
Advantages of laser excitation source
1. Laser excitation produces a high density plasma and is used for the spectrochemical
analysis of solid materials.
2. The localization effect permits examination of areas as small as 50 m in diameter,

providing the biological researcher with a tool capable of examining the insides of
individual cells without destruction of organic materials.
3. In laser excitation, the sample needs not to be electrically conducting.
39
Quantitative analysis
Use of an internal standard
If the composition of sample and matrix is unknown. The internal standard is added to
both unknown and calibration standards.
The internal standard should

1. resemble the element to be determined in rate of volatilization and chemical reactivity.
2. have a measurable emission line in the same spectral vicinity as the sample emission
line.
3. It must not also present in the original sample.
Then, by plotting the ratio of intensities of the element to the internal-standard

element vs. concentration of the element, any fluctuations should be compensated for.
40
Standard Addition Method
in order to partially or wholly counteract the chemical and spectral interferences
introduced by the sample matrix.
41
Applications of AES (using non-flame excitation sources)
AES is rapid method for qualitative and quantitative determination of most metals.
It is superior than flame and atomic absorption methods. Flame emission spectroscopy
has the limitations of being only good for few elements while atomic absorption
techniques need a separate source lamp for each element. AES methods; being very
sensitive, have numerious applications in analysis of biological samples.
For examples:
1. evaluation of platinum in body fluids and tissues after administration of platinum

containing anticancer drugs
2. determination of organic and inorganic Se compounds in biological fluids and
environmental samples
3. determination of trace elements such as Cd, Co, Cr, Cu, Fe, Hg, Mn, Ni and Pb
4. Silicon is recogonized as an essential trace element participating in normal body
metabolism. 42
SUMMARY
43
SUMMARY: Contd---
44
Thanks for your kind attention
45

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Course Code : PDA302T

By the end of this session, students will be able to:

1. Atomic Emission Spectroscopy (Flame Photometry)

Ground state atom (stable or

Spectral resonance line (The

~0% population Excited state atoms

~100% population Ground state atoms

Flame : is the source of excitation energy. (low energy source).

Events occuring in FES

Solution of metal Ground state atoms

Solvent evaporation Emission

Flame Structure Secondary combustion

Fuel- oxidant mixture

Types of burner system

Suffers from explosion hazards Aspirator air Mixing

1. Low accuracy 2. Low precision

Detectors Films or photomultipliers

Advantages and disadvantages

Hollow Cathod lamp Inert carrier gas

Total amount of light absorbed = (e2/mc2)Nf

Total amount of light absorbed = constant x Nf

Types of chemical interferences

Examples 1. Detn. of Ca in presence of sulphate or phosphate

3. Solvent extraction of the sample or of the interferring elements

Overcome: 1. by continuous calibration

Advantages of AAS: Very sensitive.

Disadvantages of AAS: Hollow cathode lamp for each element.

2. Depends only on the number of 2. Depends only on the number of

3. Absorption intensity is NOT affected 3. Emission intensity is greatly affected

1. Excellent method for trace element analysis at ppm level

High energy excitation sources

3. The main types of argon plasma sources

Argon tangential flow

The coil is energized by a radio frequency AC generator creating a changeable magnetic

Argon is not a conductor at low temperatures, but becomes electrically conducting by

It is used for multi-element determination

2- It is suitable for quantitative multielement determinations

3- The high temperature of plasma eleminates many chemical interferences present

5- The emission intensity-versus-cencentration range is linear over a very wide

2. The localization effect permits examination of areas as small as 50 m in diameter,

3. In laser excitation, the sample needs not to be electrically conducting.

Use of an internal standard

The internal standard should

Then, by plotting the ratio of intensities of the element to the internal-standard

1. evaluation of platinum in body fluids and tissues after administration of platinum

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