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N - AAS: 3.1 Limitations of Flame AAS
N - AAS: 3.1 Limitations of Flame AAS
Flame AAS was developed in the 1950s, and at this time, the environmental significance of ug/L
levels of metals such as lead and mercury, was not really appreciated. However, this would soon
change, and improvements in sensitivity were an ongoing part of instrumental development (think
about the changes in polarographic instrumentation). A number of factors limit the sensitivity of
flame AAS.
To achieve sensitivity improvements of 100 times or more (to get accurate analysis of 10 ug/L levels)
from atomic absorption spectrophotometry would require a considerably different approach: a
system AA system based around a flame and nebuliser will never achieve the sensitivities required.
There are two very different approaches, both of which dispense with the flame:
vapour generation – where the analyte is converted into a volatile form
electrothermal – where electrical heating is used
3.3 Equipment
The burner/nebuliser in FAAS instruments is removed and the electrothermal workhead put in its
place. Most modern instruments allow the two units to be interchangeable, because the rest of the
instrument is identical:
hollow cathode lamp
monochromator
photomultiplier detector
signal processing and readout
Electrothermal-capable instruments must have computer control to automate the sample delivery,
and to monitor the heating and signal measurement.
The workhead
The workhead contains the sample cell and provides the services for controlled heating of the
sample. The sample cell is a hollow graphite tube approximately 28 mm in length by 8mm in
diameter. The other common name for EAAS is graphite furnace AAS, deriving from the graphite
tube in which the sample is heated. The interior of the cylinder is coated with a layer of pyrolytic
graphite, which has a greater resistance to heat than normal graphite. The sample is delivered into
the graphite tube through a small hole in the barrel of the tube, as seen in
Figure 3.1.
There are two types of graphite tubes – partition and platform. The former
has raised sections one-third of the length along from each end. These act like
dam walls, preventing the droplet of sample spreading out too far. The
platform is a separate piece of pyrolytic graphite sitting in the centre of tube.
The droplet sits in this and supposedly is vaporised more predictably, because
the heating only occurs by radiation (the gas inside) than by conduction (from
the walls).
FIGURE 3.1 The tube is positioned inside the workhead so that the delivery hole is
Graphite furnace pointing upwards. It rests on electrodes which are connected to a low-voltage
high current power supply which delivers upwards of 3.5kW to the cylinder
AIT 3.2
3. Non-flame AAS
walls. Because graphite is only a moderate conductor of electricity, the tube heats up, providing the
conditions to vaporise and atomise the sample.
The metal housing around the furnace is water cooled to enable rapid restoration of the
furnace to ambient temperature after atomisation. Inert gas (generally argon) is flushed through and
around the tube from the ends. This has three purposes:
to remove steam and smoke vaporised during the heating
to prevent oxidation of the graphite
assist with cooling
HCL beam
quartz windows
Autosamplers
Modern electrothermal atomisation units almost always use an autosampler to place the sample in
the graphite tube. This is because the volumes involved are typically 5-20 uL, and therefore manual
loading of graphite tubes (even with micropipettes) would be unreliable, with precision being greatly
reduced compared to that obtainable with autosamplers. These also allow multiple samples to be
run without an operator in attendance.
Frequently, different solutions (blank, sample, standard, matrix modifier) are used in
combination for the analysis. These are kept apart in the plastic tubing by air gaps. The sampler arm
then swings over to the workhead and delivers the sample to the graphite tube.
AIT 3.3
3. Non-flame AAS
Therefore, the heating process must occur gradually, and involves a number of carefully programmed
steps, such as those shown in Table 3.2 and Figure 3.5:
1. Drying – removal of all volatiles at temperatures slightly above the boiling point of the solvent,
and for 30-60 seconds
2. Ashing – the temperature is increased to 500-1000C and the organic matter is burnt away,
like in a muffle furnace, for 5-40 seconds
3. Atomisation – now that all the non-elemental matter is gone, the analyte and any other
inorganic elements in the matrix are heated to the temperatures used in FAAS (1800-3000C)
for 3-4 seconds; measurement of absorbance occurs here
Analyte Matrix
Drying time No
Ashing time
Ashing temperature
Atomisation time
Atomisation temperature
AIT 3.4
3. Non-flame AAS
These notes are intended only give some general guidelines about the effect on the temperature
program of the analyte, but more importantly, the sample matrix. When setting up a method for a
new sample, it is very much a matter of trial and error until the most efficient sequence of times and
temperatures is determined.
Drying stage
This step is designed to remove all volatiles without losing analyte by over-vigorous boiling, which
could cause the mixture to splatter.
List some factors that would influence the time required for drying.
The main problem likely in the drying stage is incomplete drying which can lead to splattering of the
remaining material as the temperature increases to ashing point. You will hear the fizz as the
temperature goes up.
(b) milk
AIT 3.5
3. Non-flame AAS
Ashing stage
This step is designed to remove the non-volatile organic components of the matrix by burning them
away as in a muffle furnace.
List some factors that would influence the time required for ashing.
The ashing stage will normally be divided up into at least three steps:
increase from the final drying temperature to the ashing temperature (normally 5-10 seconds)
remain at ashing temperature for required time
in the last few seconds before atomisation, the argon gas flow is turned off so that the
atmosphere inside the tube is still for atomisation
Chemical modifiers are used where the analyte is relatively volatile and would be lost during the
ashing stage, eg lead which is volatile above 500ºC. If a modifier wasn’t used, then the ashing
temperature would have to be quite low, meaning a much longer time would be required. The
modifier for lead is phosphate, since lead phosphate has a much higher boiling point than other lead
salts, such as chloride and nitrate. However, the modifier must not hold onto the analyte during
atomisation!
Some of the volatile elements are lead, cadmium, mercury (of course), antimony and arsenic.
The instrument documentation will indicate whether a modifier is needed.
The main problem encountered with incomplete ashing is the generation of smoke during
atomisation, which causes a high background reading due to scattering. You can observe this at the
start of the atomisation step as a puff of smoke coming out through the delivery hole.
AIT 3.6
3. Non-flame AAS
Atomisation stage
This step is designed to drive the analyte atoms into an atomic vapour in the path of the HCL beam.
List some factors that would influence the temperature required for atomisation.
The argon gas is turned on after the brief atomisation step to flush the tube and avoid it turning into
CO2.
3.5 Signal measurement
This is much more complex than in flame AAS, because the analyte atoms are only present for a few
seconds once per run, rather than being present whenever the nebuliser is in the flask. Therefore,
the signal is a short-lived peak (like in a GC or HPLC), as in Figure 3.6.
This means that signal capture by the computer is essential. The alternative is to wait through
two minutes of drying and ashing and then make sure you are watching when the atomisation step
occurs. To make matters more difficult, the steam and ash generated in the first two stages cause
scattering of the beam, and an apparent absorbance which varies over time, and has nothing to do
with the real analyte absorbance. In the software, this is avoided by not having the detector
responding until the atomisation steps.
EAAS
Absorbance
FAAS
Time
AIT 3.7
3. Non-flame AAS
0.0044ms
A exp Eqn 3.1
mc
where mc is the characteristic mass, most often in pg, ms the mass of analyte in the sample/standard
(in the same units) and As the expected absorbance of the standard.
Calculating masses in pg (10-12 g) from uL and ug/L might seem a good candidate for getting
the decimal place wrong, but in fact, nothing could be easier, as shown in Equation 3.2: no unit
conversion required at all!
pg uL x ug / L Eqn 3.2
Memory effects
If a metallic element is present in the analysed aliquot at concentrations above 250 ug/L, the
atomisation stage will not be sufficient to atomise and then remove all of it. This will occur whether
the species is the analyte or in the matrix. If it is the analyte, then some proportion will remain in the
tube when the next run commences. This will produce a higher absorbance than it should, and is
known as a memory effect.
AIT 3.8
3. Non-flame AAS
The solution to this problem is a number of tube cleans, which take the temperature up to above
2000C very rapidly. Depending on the element and the concentration, it may require three or four
tube cleans before continuing.
Solutions
A typical analysis run may use at least three different solutions: sample, added standard, blank,
modifier. The total volume of liquid should be between 15 and 30 uL, and should be consistent for all
solutions.
Tube performance
The graphite tube has a finite lifetime of around 500 normal temperature cycles. This will be reduced
by various factors.
Background correction
Background correction is a means of correcting for sources of non-analyte reduction in beam
intensity. These sources include:
smoke particles
molecular species
These are more significant (especially the smoke) in EAAS than FAAS. When it occurs, the measured
absorbance is the sum of analyte and background absorbance (as in Eqn 3.3). An ideal system would
measure the total absorbance, and also the background by itself.
At = Aa + Ab Eqn 3.3
AIT 3.9
3. Non-flame AAS
Hollow Monochromator
Cathode & Detector
Lamp
Reaction vessel
(sample + reagents)
In the case of mercury, the analyte is already in the chemical form required – Hg(g) – after chemical
treatment in the reaction vessel by reduction of ionic mercury by tin chloride, so no heating of the
quartz tube is required, hence the name “cold vapour”.
AIT 3.10
3. Non-flame AAS
In the hydride generation method, the analyte is reduced, usually by sodium borohydride (NaBH4),
producing the molecular hydride. When this is flushed out in the gas stream, it is not in the atomic
form, and requiring heating in the tube to decompose the hydride into the atomic metal. This
procedure is applicable to As, Se, Te, Sb, Bi and Sn. The tube may be heated electrically or with a
flame.
Revision Questions
1. (a) Give THREE factors that limit the sensitivity of flame AAS.
(b) Explain how electrothermal AAS overcomes these.
2. What services other than electricity are required for EAAS? What function(s) do they serve?
3. Why is an autosampler an essential part of an EAAS instrument?
4. Why is the gas switched off at the end of the ashing stage?
5. What changes would you make to the basic “analyte in water” temperature program for the
following analyses:
(a) aluminium in milk
(b) lead in petrol
6. Why is a phosphate modifier recommended for lead analyses? How does it work?
7. The characteristic mass for aluminium is 6.1 pg. What is the expected absorbance for a 5 uL
aliquot of 50 ug/L standard?
8. What type of samples would most require background correction?
9. Explain the following for measurements of lead in a wine sample. Using the standard
temperature program for lead in water, the sample gives an absorbance of 0.6
(a) When the ashing time is increased from 5 to 60 seconds, the absorbance decreases to 0.3.
(b) When the drying time is reduced from 60 seconds to 5, the absorbance decreases to 0.1.
(c) When the modifier is not used, there is no absorbance.
10. Why is tin chloride required in mercury analysis by vapour generation AAS?
11. Why is heat needed in the vapour generation analysis of arsenic?
AIT 3.11
3. Non-flame AAS
1. (a) inefficiency of nebuliser, reaction in the flame, formation of oxides, background colour of
flame
(b) Table 3.1
2. p3.3
3. p3.4
4. top of p3.7
5. (a) increase drying & ashing times (b) decrease drying temp, small increase in ashing time
6. p3.7
8. High organic content, high levels of elements giving molecular species eg Al, Ca
9. (a) significant amounts of smoke in the standard conditions run causing higher absorbance,
this is removed during the longer ashing
(b) insufficient drying time means that when the temperature goes up to the ashing point, the
remaining moisture is rapidly lost, causing the sample to fizz and spray out of the tube.
(c) the lead becomes volatile during the ashing stage and is lost before measurement
10. For this analysis, mercury must be in the elemental form (Hg0) so the tin chloride reduces the
+1 and +2 ionic forms.
11. The volatile form of arsenic is AsH3, which must be decomposed (by heat) to form As to absorb
the HCL radiation.
AIT 3.12