Atomic Absorption / Emission / Fluorescence

Spectroscopy
Atomic Absorption Spectroscopy
The analyte concentration is determined from the
amount of absorption.
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Atomic Absorption Spectroscopy
It is possible to measure the concentration of an
absorbing species in a sample by applying the Beer-
Lambert Law:


Abs = log
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Abs = ccb
c = extinction coefficient
Atomic Absorption Spectroscopy
But what if c is unknown?
Concentration measurements can be made from
a working curve after calibrating the instrument
with standards of known concentration.
Atomic Absorption Spectroscopy
Instrumentation

Light Sources

Atomization

Detection Methods
Light Sources
Hollow-Cathode Lamps (most common).

Lasers (more specialized).

Hollow-cathode lamps can be used to detect one
or several atomic species simultaneously. Lasers,
while more sensitive, have the disadvantage that
they can detect only one element at a time.

Hollow-Cathode Lamps
It is important to note that atoms producing an emission line in a
hollow cathode lamp are relatively at a lower temperature than
those in a flame. As a result emission lines in a HCL lamps are
broadened less than that the emission lines in flames.
A broadened emission line of a hollow cathode lamp (1) and a broadened
emission line of a flame (2)
Atomization
Atomic Absorption Spectroscopy (AAS) requires that
the analyte atoms be in the gas phase.


Vaporization is usually performed by:
Flames
Furnaces
Plasmas
Flame Atomization
Flame AAS
can only
analyse
solutions.
Flame Atomization
Degree of atomization is temperature
dependent.

Vary flame temperature by fuel/oxidant
mixture.
Fuel Oxidant Temperature (K)
Acetylene Air 2,400 - 2,700
Acetylene Nitrous Oxide 2,900 - 3,100
Acetylene Oxygen 3,300 - 3,400
Hydrogen Air 2,300 - 2,400
Hydrogen Oxygen 2,800 - 3,000
Cyanogen Oxygen 4,800
The
Atomization
process in a
Flame

Furnaces
Electro-thermal Atomizers
It gives an enhancement of sensitivity by 20 to 1000 times
compared to flame technique, as the entire sample is
atomized in a short period and the average residence time of
the atoms in the optical path is a second or more.

The commercial tubes are composed of graphite with
relatively impervious pyrographite coating.

The tube is resistively heated by an external power supply
and is protected from oxidation by a flow of sheath gas
typically argon around the furnace.

The heating of the tube containing the sample is carried out
through the three steps: drying, ashing and atomizing of the
analyte sample.
Furnaces
Inductively Coupled Plasmas
Enables much higher temperatures to be achieved.
Uses Argon gas to generate the plasma.

Temps ~ 6,000-10,000 K.


Used for emission experiments rather than absorption
experiments due to the higher sensitivity and elevated
temperatures.

Atoms are generated in excited states and
spontaneously emit light.
Detection
Photomultiplier Tube (PMT).

Cold Vapor Atomic Absorption Spectrometry.
Hydride Generation
Atomic Absorption Spectroscopy

Calibration of Laboratory Instruments
Initial Calibration

Either analyst prepare the standard or use commercial ones.

The concentrations of the standards must be within the optimum
range of that given by the method.

Every standards must have an expiry date & it should not be used
beyond that.

The number of standards needed is recommended by the METHOD
or the MANUFACTURER of the method.

When not given use a minimum of three concentration and a blank.
However six standards are more than enough.

The 3 Standards are selected as follows

The concentration of the high standard is the upper level of
the optimum range.

The concentration of the middle standards is the of the
highest standard.

The value of the lower standard is 1/5 of the highest standard

Plot the calibration curve & this must be approved using the
corresponding correlation coefficient

Using the regression analysis
Corr. Coeff. > 0.9998
If not satisfied recalibrate.
Calibration Verification Standards (CVS)
It is a standard (with a known value) prepared
from a source other than that used to prepare
the calibration standard.

e.g. Certified Reference Materials (CRM). The
value obtained from calibration curve cannot be
deviated by no more than 10%.
Optimum Concentration Range

The concentration range where the calibration curve is linear.

It is defined by the method and varies with the instrument

Reagent Blank
Analytical free water analyzed with samples.

Method Blank
Analyte free water that is subjected to the same pretreatment
as samples.
Spike sample or Matrix spike
How are they used?
A small quantity of a known concentration of analyte stock solution is added to
the sample.

If the analyte sample is a liquid thoroughly mix after adding the stock standard.

Solid sample are mixed thoroughly in a glass/ceramic bowl before taking
aliqnot out of it.

The amount of spike should produce a signal at least 10 times that equivalent
to IDL and it should not produce a signal larger than 100 times equivalent to
IDL.

The concentration of the standard added generally should be 3 to 5 times the
analyte expected in the sample.

Spike solution must be added before the sample preparation.

Detection limits
Instrument Detection Limit (IDL)

Measure instrument responds for several aliquots
of analyte free water ( e.g. Number of aliquots = 7).

calculate the standard division, S for all the
response

IDL = 3 S

IDL varies with the analyte for a given instrument
Method Detection Limit (MDL)
prepare a solution using analyte and analyte free water having a
concentration near the quoted minimum detectable concentration,
Divide the solution in to several portions (e.g. 7).

Measure instrument response for each sample, the response for the
analyte. Use a student ttable to determine tvalue for the 7 replicate
measurements, with the required level of confidence.

MDL = t S
e.g. for the above measurements of degrees of freedom (n 1) = 7-1 =
6

If a confidence limit of 98% is required, t=3.14
MDL = 3.14 S
Practical QuantitationLimits (PQL) or
Limit of Quantitation(LOQ) or Limit of
Reporting ( LOR)


All these 3 determinations are the same parameter.
The limit of detection ( LOD) are good enough one
to be sure that an analyte is present or absent. If
response is larger the MDL or IDL then the analyte
is present. However, its value can not be accurately
reported as the concentration is low.

PQL = LOQ = LOR = 10 s

Accuracy:
some times represented by the quantity called
percentage recovery (%R) defined as below

Precision