AAS .PPT (Compatibility Mode)

ATOMIC ABSORPTION
SPECTROSCOPY
CONTENTS:
n Principle of AAS.
n Instrumentation.
n Applications.
n Experiments.
INTRODUCTION:
n Atomic Absorption Spectroscopy is a very common

technique for detecting metals and metalloids in
samples.
n It is very reliable and simple to use.
n It can analyze over 62 elements.
n It also measures the concentration of metals in the

sample.
HISTORY:
The first atomic absorption spectrometer was built by

CSIRO scientist Alan Walsh in 1954. Shown in the
picture Alan Walsh(left), with a spectrometer.
Elements detectable by atomic absorption are highlighted in pink in this
periodic table
PRINCIPLE:
n The technique uses basically the principle that free

atoms (gas) generated in an atomizer can absorb
radiation at specific frequency.
n Atomic-absorption spectroscopy quantifies the

absorption of ground state atoms in the gaseous state .
n The atoms absorb ultraviolet or visible light and make

transitions to higher electronic energy levels. The
analyte concentration is determined from the amount of
absorption.
- Concentration measurements are usually determined from a
working curve after calibrating the instrument with
standards of known concentration.
- Atomic absorption is a very common technique for

detecting metals and metalloids in environmental samples .
THEORY:
Hollow Cathode
Lamp
Detector Nebulizer
Atomic
Absorption
spectrometer
Monochromator Atomizer
Schematic diagram of AAS:
LIGHT SOURCE:
n Hollow Cathode Lamp are the most common radiation

source in AAS.
n It contains a tungsten anode and a hollow cylindrical

cathode made of the element to be determined.
n These are sealed in a glass tube filled with an inert gas

(neon or argon ) .
n Each element has its own unique lamp which must be

used for that analysis .
Hollow Cathode Lamp:
Quartz window
Pyrex body
cathode
Anode
Cathode
NEBULIZER:
n suck up liquid samples at controlled rate.
n create a fine aerosol spray for introduction into flame.
n Mix the aerosol and fuel and oxidant thoroughly

for introduction into flame.
Atomizer
n Elements to be analyzed needs to be in atomic sate.
n Atomization is separation of particles into individual

molecules and breaking molecules into atoms. This is
done by exposing the analyte to high temperatures in a
flame or graphite furnace .
ATOMIZERS:
ATOMIZER
FLAME GRAPHITE TUBE

ATOMIZERS ATOMIZERS
FLAME ATOMIZER:
n To create flame, we need to mix an oxidant gas and a

fuel gas.
n in most of the cases air-acetylene flame or nitrous oxide-

acetylene flame is used.
n liquid or dissolved samples are typically used with flame

atomizer.
GRAPHITE TUBE ATOMIZER:
n uses a graphite coated furnace to vaporize the sample.
n ln GFAAS sample, samples are deposited in a small

graphite coated tube which can then be heated to
vaporize and atomize the analyte.
n The graphite tubes are heated using a high current

power supply.
MONOCHROMATOR:
n This is a very important part in an AA spectrometer. It is

used to separate out all of the thousands of lines.
n A monochromator is used to select the specific

wavelength of light which is absorbed by the sample,
and to exclude other wavelengths.
n The selection of the specific light allows the

determination of the selected element in the presence of
others.
DETECTOR:
n The light selected by the monochromator is directed

onto a detector that is typically a photomultiplier tube ,
whose function is to convert the light signal into an
electrical signal proportional to the light intensity.
n The processing of electrical signal is fulfilled by a signal

amplifier . The signal could be displayed for readout , or
further fed into a data station for printout by the
requested format.
Calibration Curve
n A calibration curve is used to determine the unknown
concentration of an element in a solution. The
instrument is calibrated using several solutions of known
concentrations. The absorbance of each known solution
is measured and then a calibration curve of
concentration vs absorbance is plotted.
n The sample solution is fed into the instrument, and the

absorbance of the element in this solution is measured
.The unknown concentration of the element is then
calculated from the calibration curve
APPLICATIONS:
Determination of even small amounts of metals (lead,

mercury, calcium, magnesium, etc) as follows:
n Environmental studies: drinking water, ocean water, soil.
n Food industry.
n Pharmaceutical industry.
Guidelines for
Trouble Shooting
and Maintenance of
AA Systems
AA Maintenance & Trouble Shooting Feb. 2014

Common AA Problems Reported by Customers
Sensitivity:
• Sensitivity is worse than it used to be
• I have a new application and I can’t get the sensitivity I need
• How come I can’t get the instrument to meet published detection limits?
Precision
• Sensitivity is acceptable but precision is terrible
High noise
• Can get the “right answers”, but very noisy signal – this is also giving bad precision.
Accuracy
• Instrument does not give the “right” results.
Poor Sample Throughput

• The instrument throughput needs to improve
• Burner blocks too quickly
2
Causes of Poor Flame AA Sensitivity
Flame Blocked nebulizer

atomization Dirty burner (reducing pathlength)
system
Broken impact bead
Poor optimization
Optimization Optics setting – using right wavelength/slit?
Wrong burner type
Interferences
High blank level
Standard
(& sample) Standards prepared correctly?
preparation
Samples prepared correctly? – ionization suppressant
Low acetylene gas pressure – acetone carryover
3
Causes of Poor Flame AA Precision
Dirty burner (“ragged”
Flame
atomization
flame) Impact bead
system
adjustment Impact bead
condition
Not fitting mixing
Optimization
paddles Burner
alignment
Poor optimization
Standard
(& sample) High nebulizer flow rate
preparation
Gas purity
Wash-out (memory effects) 4

Causes of High Noise in Flame AA
Dirty burner (“ragged” flame)
Flame
atomization Impact bead adjustment
system Impact bead condition
Not fitting mixing paddles
Burner alignment
Optimization Poor optimization – especially the HC lamp
Wrong lamp operating current
High nebulizer flow rate
Dirty optics
Gas purity
Standard
(& sample) Incomplete digestion – particles in solution
preparation
5
Flame Atomization System Tips
Do: Don’t:
Check optimization each analysis
Assume system is still optimized
Check/monitor the nebulizer uptake
Assume nebulizer flow rate is the same
Check/adjust the impact bead
Check the blank reading
Rinse between samples & at the end of Use a simple water blank
the run
- Rinse should match sample matrix
Clean the burner/spray chamber Wait until you have blockage before
regularly cleaning
- Inspect condition of the impact bead
Follow analytical recommendations in
“cookbook”
6
Burner Alignment Tips
Burner
• Burner position must be optimized
(vertical, horizontal & rotational
positioning)
• Use “target area” on burner alignment

card to ensure light beam runs
parallel to burner slot
Optimum viewing height for Ca

• Adjust burner height while aspirating a
standard and optimize for maximum
signal
- Optimum height varies depending on flame
chemistry
• Burner type changes path length

(for air/acetylene elements)
7
Optimization of the Nitrous Oxide/Acetylene Flame
Lean Stoichiometric Rich

Reduce acetylene flow Add extra acetylene
8
Cleaning the Atomization System
Cleaning the burner:

• Polish with metal polish like “Brasso”
- Watch the video (#4)
Cleaning the spray chamber:

• Wash in detergent solution
• Check the impact bead condition & setting
- Replace if badly pitted
Removing nebulizer blockage:

• Disassemble completely and wash in detergent
• Reassemble and test
9
Impact Bead Setting
Loosen bead securing screw

Turn external adjuster fully
clockwise
Set the bead position
• 2 critical settings
- Bead should be centred in front of
venturi
- Distance between bead & venturi
should be ~0.1mm (thickness of
sheet of paper)
Tighten adjuster to lock bead in

place
10
Flame Atomization – Rec. Settings
For most flame AA applications:
• Fit the mixing paddles
• Optimize the impact bead for best sensitivity
• Use narrow bore capillary tubing
For highest flame AA sensitivity:

• Remove mixing paddles
• Adjust impact bead further away from the nebulizer
• Use wide bore capillary tubing (highest uptake rate)
For higher TDS samples:

• Fit the mixing paddles
• Adjust impact bead ½ to 1 turn clockwise (towards nebulizer) from optimum sensitivity position
• Use wide bore capillary tubing (to reduce chance of blockage)
11
Recommended Procedures at End of the day
1. Aspirate distilled water for a few minutes before shutting off flame
2. Allow burner to cool
3. Remove burner and clean by running water through it
4. Dry burner by shaking
5. Pour 500 mL of water into spray chamber, through burner socket
6. Replace burner
7. Empty waste vessel
12
Flame AA Performance – Benefits
Flexibility, Ease of Use & Superior Flame Performance
Tunable performance means…

• Highest flame sensitivity:
> 0.9 Abs. for 5 mg/L Cu
• Best precision: < 0.5 % RSD
using 10 x 5 s readings
• Extended operation with
difficult samples
• No loose gas hoses and
no tools required for gas
connection
• Fast change-over to furnace
operation (manual - < 30 s)
13
Causes of Poor Furnace AA Sensitivity
Missed injection
Furnace
workhead Aged (or damaged) tube in use
Wrong electrodes fitted
Poor optimization – esp. drying conditions
Workhead incorrectly aligned
Optimization
Optics setting – using right wavelength/slit?
No modifier (or incorrect) modifier used
Use of nitrogen as inert gas
No acid in solution
Standard High blank level
(& sample)
preparation Standards prepared & stored correctly?
Samples prepared correctly – digestion?
14
Causes of Poor Furnace AA Precision
Missed injection
Bubble formation in syringe
Furnace
workhead
Dirty dispensing capillary
Using non-Agilent graphite tubes
Graphite components excessively worn – poor electrical contact
Wrong dispensing height
Optimization
Poor optimization – esp. drying conditions
Missing a cooldown step (esp. with platforms)
Gas purity
No acid or detergent in rinse (memory effects)
Standard
(& sample) No acid in solution
preparation Incomplete digestion – particles in solution
15
Causes of High Noise in Furnace AA
Furnace Dirty windows in workhead

workead
Use of platforms
High background levels
High absorbance signals
Optimization
Wrong lamp operating current
Workhead incorrectly aligned
Poor optimization – especially the HC lamp
Dirty optics
Gas purity
Standard
(& sample) Incomplete digestion – particles in solution
preparation
16
Furnace AA System Tips
Do: Don’t:
Check optimization each analysis Assume system is still optimized
Check/monitor the dispensing height Assume dispensing height is the same
Ensure the rinse solution has 10 drops Use a simple water rinse
conc. HNO3 + 5 drops Triton X-100
Remove residue from the dispensing Start analysis with a dirty capillary tip
capillary
Check/monitor the graphite tube Start analysis with a tube near the end
Check the blank reading of

it’s life
Clean the workhead regularly
- Inspect condition of the graphite components
Follow analytical recommendations in
“cookbook”
17
Furnace AA System Tips - Workhead Alignment
Furnace workhead
• Workhead position must be optimized
(want light beam to pass through centre of
graphite tube)
- Align lamp first (no workhead),
then place workhead in position and align
Sample Dispenser settings

• Carefully adjust injection depth – easy with the furnace camera
Light Beam Aligned

Through Center of
Graphite Tube
18
Furnace AA System Tips - Setting Injection Depth
Correct dispensing
Capillary tip must remain in contact
with solution during dispensing
Reduce dispensing height if sample

spreads due to low surface tension þ
Ensure there is no liquid on the
outside of the capillary after
Pipet tip too low!
dispensing
Ensure there is no liquid inside the

ý
capillary tip after dispensing
Pipet tip too high!
ý
Sample should remain as a drop in
the centre of the tube
19
Furnace AA System Tips – Tube Conditioning
Why condition the tubes before use?
• Helps remove residual contamination
• Gently “beds” a new tube in
- Important when determining concentrations near detection limit
- Also important with some complex matrices
• Critical when using modifiers
- Helps to build up coating inside the tube
- Improves efficiency of the modifier
• Improves reproducibility
Recommended process
• Use “Tube Condition” facility (or otherwise, fire tube 5-10 times using
either reagent blank or representative sample)
20
Furnace AA System Tips – Method Parameters
What to Check?
- Furnace parameters
• Set appropriate drying temperature and time (2-3 sec/uL of solution injected)
• Optimize ashing temperature using ashing study – use SRM optimization
• Ensure inner gas flow “off” just prior to atomization
Does the sample sizzle or

splatter during the dry stage?
• Listen for the

sound
• Use the mirror or
furnace video to monitor
the sample drying
21
Furnace AA System Tips – Method Optimization
• SRM “Wizard” automates furnace
optimization
• Optimizes absorbance as a function of

ashing and atomization temperature
• Automatically creates a method using

recommended conditions
• Reduces training requirements for new

users
Optimization
results for Pb
determination
using phosphate
modifier
22
Furnace AA System Tips – Reducing Sensitivity
May be required due to sensitivity of this technique:
• Switch to an alternate wavelength

- Select a less sensitive wavelength (if available)
• Reduce sample volume
• Use slower ramp rate to atomization
- Aim to broaden the peak during atomization
• Use low gas flow during the atomization step
23
Furnace AA System Tips – Factors Influencing Tube
Lifetimes
Graphite components excessively worn – poor electrical contact
No tube preconditioning (or always using Tube Clean)
Sample matrix
Inert gas used
• Argon gives longest life
• Nitrogen degrades tube faster due to oxygen presence
Temperature program used Type Zeeman Tube Lifetime
30 0.20
of chemical modifiers used 25
0.18
CCV Concentration ug/L

0.16
• Powerful oxidizing agents degrade tube faster 0.14
Absorbance
20 Firings Conc
0.12
Abs 2.453501
- Perchloric acid 15 CCV
CCV
209
CCV Concentration
212
512 25.4518 0.10
520 25.4837 0.08
0.1492
0.1495
CCV Abs orbance
- Perchlorates 10 CCV
CCV
215
218
527 25.3244
534 25.4040
0.1486
0.06
0.1490
CCV 221 542 25.7384 0.040.1508
- Sodium nitrate 5
CCV 224 549 25.4200 0.020.1491
CCV 225 552 25.6141 0.000.1499
0
CCV 228 559 25.7588 0.1507
0 1000 2000 3000 4000 5000
CCV 231 566 25.6141 0.1498
Number of Firings
CCV 234 574 25.7588 0.1506
24
Tips to Improve Standard Preparation
- How are they prepared?
• Ensure purchased standards are still within “Use By” date
• Use calibrated pipettes and class ‘A’ volumetric flasks for dilutions
- Periodically, check accuracy & reproducibility of your pipettes
• Use de-ionized water (Type I - conductivity > 18 Md/cm3)
- Lower grades may have contamination
• Use serial dilutions for preparing low concentrations from 1,000 ppm stock
- Please don’t do large dilutions (> 1:10,000) in 1 step
- What concentration are they?

• Low concentration standards have a finite life
- Prepare ppb (ug/L) concentration standards daily from high conc. stock
- Prepare low ppm (mg/L) concentration standards weekly
- How are they stored?

• Plastic vessels ensure better stability
• Stabilize with acid – low pH ensures better stability
25
Tips to Reduce Contamination
Contamination can come from anything that comes into
contact with your sample during storage, digestion
(dilution) and analysis
• Check reagent purity
- Always buy the best reagents
- Always check the certificate of analysis for
elevated levels
- Caution if buying in large quantities
• Worst case – can use contaminated acid for cleaning
• Ensure still within “use by” date
- Reseal immediately after use
• Other common contamination sources

- Reagent water
- Clean glassware?
- Airborne dust in the lab.
- Pipette tips
• Don’t insert pipette tips into your acids
• Use natural tips – colored tips may increase
contamination (Cu, Fe, Zn, Cd)
- Powdered gloves (esp. for Zn)
26
Tips to Improve Accuracy of Results
- Sample preparation
• Is the most appropriate digestion being used?
• Are all of the analytes being quantitatively (and reproducibly) extracted and
dissolved?
- Many digestions are only partial extracts – efficiency will vary with the sample
matrix
- Some volatile analytes may be “lost” during digestion
• Confirm by taking a solid certified reference material through your preparation and
analysis procedure
• Is the digest stable – or are you seeing any precipitates or a suspension?

• Do you see any potential contamination from either reagents or the digestion
equipment? e.g. especially with Si, B or Ca
• Include a “Reagent Blank” with every sample batch to monitor
27
Tips for Cleaning Dirty Optics
Monitor the windows regularly

- Check lamp for fingerprints
- Check sample compartment windows for build-up
of film/chemical residue
Smudges or chemical residue reduces throughput

light and increases noise
Cleaning the windows?

- Wipe clean with an optical tissue
(as you would use to clean a camera lens) Cleaning end windows
- If necessary, use optical tissue moistened with ethanol from furnace
workhead
28
Flame AA – Recommended Maintenance Schedule
Daily:
• Check the gas delivery pressures & cylinder contents (esp. acetylene)
• Check exhaust system
• Check the nebulizer uptake rate and burner condition
• Clean burner compartment & instrument
• Empty waste container
Weekly:
• Clean burner (or earlier if required)
• Disassemble flame atomization system and clean
- Check condition of O rings and impact bead (no pitting)
Monthly:
• Check windows in sample compartment (clean if necessary)
29
Furnace AA – Rec. Maintenance Schedule
Daily:
• Check the gas delivery pressures & cylinder contents
• Check exhaust system
• Check condition of the graphite tube – replace as necessary
- When replacing the tube, check the condition of the electrodes
• Check dispensing capillary “free” and syringe
• Top up rinse reservoir as required
• Clean the workhead around the sample injection hole
• Empty waste container
Weekly:
• Check furnace workhead windows (clean if necessary)
30

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ATOMIC ABSORPTION

n Atomic Absorption Spectroscopy is a very common

n It is very reliable and simple to use.

n It can analyze over 62 elements.

n It also measures the concentration of metals in the

The first atomic absorption spectrometer was built by

n The technique uses basically the principle that free

n Atomic-absorption spectroscopy quantifies the

n The atoms absorb ultraviolet or visible light and make

- Atomic absorption is a very common technique for

n Hollow Cathode Lamp are the most common radiation

n It contains a tungsten anode and a hollow cylindrical

n These are sealed in a glass tube filled with an inert gas

n Each element has its own unique lamp which must be

n suck up liquid samples at controlled rate.

n create a fine aerosol spray for introduction into flame.

n Mix the aerosol and fuel and oxidant thoroughly

n Atomization is separation of particles into individual

FLAME GRAPHITE TUBE

n To create flame, we need to mix an oxidant gas and a

n in most of the cases air-acetylene flame or nitrous oxide-

n liquid or dissolved samples are typically used with flame

n uses a graphite coated furnace to vaporize the sample.

n ln GFAAS sample, samples are deposited in a small

n The graphite tubes are heated using a high current

n This is a very important part in an AA spectrometer. It is

n A monochromator is used to select the specific

n The selection of the specific light allows the

n The light selected by the monochromator is directed

n The processing of electrical signal is fulfilled by a signal

n The sample solution is fed into the instrument, and the

Determination of even small amounts of metals (lead,

n Environmental studies: drinking water, ocean water, soil.

AA Maintenance & Trouble Shooting Feb. 2014

Poor Sample Throughput

AA Maintenance & Trouble Shooting Feb. 2014

Flame Blocked nebulizer

AA Maintenance & Trouble Shooting Feb. 2014

Wash-out (memory effects) 4

AA Maintenance & Trouble Shooting Feb. 2014

• Use “target area” on burner alignment

Optimum viewing height for Ca

• Burner type changes path length

AA Maintenance & Trouble Shooting Feb. 2014

Lean Stoichiometric Rich

AA Maintenance & Trouble Shooting Feb. 2014

Cleaning the burner:

Cleaning the spray chamber:

Removing nebulizer blockage:

Loosen bead securing screw

Tighten adjuster to lock bead in

AA Maintenance & Trouble Shooting Feb. 2014

For highest flame AA sensitivity:

For higher TDS samples:

AA Maintenance & Trouble Shooting Feb. 2014

2. Allow burner to cool

3. Remove burner and clean by running water through it

4. Dry burner by shaking

5. Pour 500 mL of water into spray chamber, through burner socket

7. Empty waste vessel

AA Maintenance & Trouble Shooting Feb. 2014

Tunable performance means…

AA Maintenance & Trouble Shooting Feb. 2014