PerkinElmer AAnalyst 300

Location: Friedland 202

PerkinElmerAA300.jpg:

How It Works and What It Does

The AA-300 is used to determine the unknown concentration of an individual element by measuring the amount of light the unknown
concentration absorbs. Instrument readouts are calibrated to display concentration directly. The basic instrumentation requires a primary light
source, an atom source and a detector. The light source is a hollow cathode lamp (HCL) or electrode discharge lamp (EDL), which emits a
spectrum specific to the element from which it is made. The light beam passes through either an air-acetylene or nitrous oxide-acetylene flame,
which has a high temperature and specific flow rate. The atom source introduces the sample into the flame as an aerosol and the heat produces
free analyte atoms. Analyte atoms in the flame that correspond to the particular lamp being used absorb energy from the light source. A detector,
usually a photomultiplier tube, measures the light intensity exiting the flame. The electronics convert the amount of light absorbed to the actual
sample concentration.

The AA-300 is equipped with an automatic six-lamp turret and is programmed to perform sequential multielement analysis. The AA-300 is
equipped with lamps for the following elements: Ag, Al, As, Ba, Ca, Cd, Cr, Cu, Fe, K, Na, Mg, Ni, Pb and Zn The Iron(Fe) and Copper(Cu) lamps
are the most commonly used.

Directions for Use

Instructions for Use

by Golda Yelaun

Instruction for AA – Flame Analyst-300.

1. Check the location of exhaust vent (should be located at least 4 meters from the Flame).
2. Check the pressure of the AA Gas Purifier assembly. Make sure the output pressure is between 51 and 65 psig. ( on the wall , behind the
instrument).

3.
 3. Turn on the proper gas supplies and set pressures, recommended in the handbook.

Acetylene 85-100 kPA (12-14 psig)

Air (dry grade) 350-450 kPA (50-65 psig)

Nitrous oxide 345-520 kpa (50-75 psig)

4. Switch on the computer and start the Windows.

5. Switch on the spectrometer and accessories.
6. Start the AAWinLab. To remove the initial screen, click on Menus and Toolbar.
7. In the File menu, click on Open – Method… In the Open Method dialog, select the method you want to open, then click on OK.
8. Connect the lamp to appropriate socket.
9. On the tool bar click on Lamps.
10. Click on the appropriate Lamp to switch it ON.
11. Wait for couple minutes to warm up, click on Midscale to set the current.
12. Close the Align Lamps window.
13. Before you ignite the flame adjust the burner Height.
a. in the Tools menu, click on Continuous Graphics.
b. Open the flame compartment door.
c. Place a small piece of white cardboard over the burner, facing the light beam. Turn the horizontal adjustment knob until the light
beam is centered over the burner slot.
d. Lower the burner with vertical adjustment knob so that the burner head well below the light beam.
e. Click on Auto zero.
14. Check if the drain system is installed and operating correctly.
15. On the Toolbar click on Flame.
16. Aspirate the blank solution(distilled water). When the signal is steady, click on Autozero in Continuous Graphics Window.
17. On the toolbar click on Manual and Results.
a. in the Manual Control Window click on Analyze Blank. (you need to place the tubing in the solution before you click on Analyze)
b. Analyze Standards.
c. Analyze Sample.

Shutting down the Instrument after Flame Analysis

by Golda Yelaun

1. With the Flame still burning aspirate deionized water for five minutes.
2. In the Flame Control window, click on Flame off.
3. Turn off the burner gas supplies to the spectrometer.
4. Exit the software, including the main application and the Windows software.
5. Switch off the Spectrometer.
6. Switch off the computer.
7. Empty the burner drain vessel. Dispose the waste solutions according the Safety Regulations.

Instrument Calibration

1. Curve correction: Quantitative measurements in Atomic Absorption is based on Beer’s Law, which states that concentration is
proportional to absorbance: C = kA. The Perkin-Elmer AA-300 allows either linear or non-linear calibration using multiple calibration
standards.
2. Selecting the number and concentration of standards: Selecting the number and the concentration of calibration standards is very
important. If the analyte concentration of all samples to be analyzed falls within the linear range, one calibration standard should be used.
The top of the linear range for the most elements is between 0.20 and 0.30 absorbance units. If the analyte concentration in the samples
is expected to exceed the linear range, either two or three calibration standards should be used.
3. How to select calibration standards properly:
a. Make up three standards: The first one (S1) should be on the top of the linear range. The concentration of the second standard
(S2) should be 3X the concentration of the first. The concentration of the third standard (S3) should be 6X the concentration of
the first standard (S1) (6:3:1 rule).
b. Preparing the calibration standards: Calibration standards are prepared from certified stock Standards for calibration. A variety of
stock standards are usually provided at concentration level at 1000mg/l, and are available directly from Perkin-Elmer as well as
from other commercial sources.
4. Creating a calibration curve:
a. Set up the spectrometer in the concentration mode. Auto zero on Blank.
b. In the Manual Analysis Control window click Analyze Blank and then Analyze Standards.
c. Click on Analyze Sample.
d. Check the calibration curve. If the calibration curve in the linear range and some standards appear to be off it, check them and
 d.

repeat the calibration.

Example: Creating a Calibration Curve for Fe (Iron)

by Golda Yelaun

1. Creating a Calibration Curve

1. Preparing the calibration standards. Calibration standards are prepared from stock solution 1000 PPM Fe (Atomic Absorption standard).
We prepared three calibrations standards: 2ppm, 5ppm, 10ppm.
2. In the Manual Analysis Control window we have to click Analyze Blank, then Analyze Standard.Since the Method is already stored all
three standards (2,5,10 ppm) will be analyzed.
3. The instrument will create the plot Absorbance versus Fe-concentration automatically.
4. The calibration curves have to be linear. If any of the standards appear to be off the calibration curve, we have to check the standards
and repeat the calibration.

2. Analyzing the samples.

Before analyzing prepared samples we have to put QC sample and Blank for checking the instrument.

1. Analyzing Reagent Blank (absolute value have to be in the Range 0.2 mg/l)
2. Analyzing QC standard- 5ppm Fe (have to be closed to 5mg/l)
3. Analyzing prepared samples.(Concentration of the samples have to be in the Linear Dinamic Range – less than 10 ppm Fe)
4. To the end of the Run (around 10 samples) to check again Reagent Blank and 5ppm Fe.

Tips For Use

1. Manuals for the AA are either in the cabinet drawers under the instrument or in Golda Yelaun's office.
2. A good description of using the AA is available in the General Chemistry Lab Manual, updated and released every year.
3. The AA is calibrated every day, since there are different standards for different elements.
4. Note!: If the AA generates an error message "Can't ignite the flame", check that the gas is not shut off on the wall behind the AA.
5. Note!: The numerical readings generated by the AA should be close together. If the numbers jump around, that indicates that the AA
should be cleaned. This might be needed after using a sample that hadn't been filtered well.
6. If the absorbance signal is low, there may be a problem with the lamp.
7. Note:!: If the AA is generating unexpected readings, check that the trap flask under the machine is not too full. If it is too full, fumes from
the material in the flask can alter the signal. Check the manual for cleaning instructions.
8. If the AA is still generating unexpected readings, run 1-1.5 liter distilled water through the trap flask tube to clean it out.
9. Also check the sample tube for blockages.
10. The compressed air must be dry grade. Humidity in the compressed air will damage the AA.
11. If the pressure level in the compressed air tank is 200 psi or less, change the tank, since it is almost emtpy.
12. If the pressure level in the acetylene tank is 10 psi or less, change the tank, since it is almost empty.
13. The AA software on the PC has the recommended conditions for burning. The different burners are in the cabinet drawers under the AA.
14. Each lamp has a readout showing usage and condition. Check this to insure that the lamp is in good working condition.
15. Some troubleshooting tips from Perkin Elmer:
a. If the results are low:
i. standard solutions may have be improperly made
ii. blank solution maybe contaminated
iii. weak lamp
b. If the results are “Noisy”:
i. burner may be contaminated. Clean the burner head and burner chamber.
ii. sample contaminated.
c. Flame doesn’t ignite :
i. Check fuel and oxidant is maintained at proper pressure
ii. make sure the drain interlock is connected
iii. the level of liquid in the drain vessel too low: pour 200-300ml of deionized water into the drain hose.
iv. the level of liquid in the drain vessel too high: empty the drain vessel
d. Lamp problems:
i. lamp will not light: lamp not plugged in; lamp is not aligned properly; lamp is old or damaged – try another lamp.
16. Contact Golda Yelaun with any questions.
Contact Point Information
1. Contact Prof. Dolnik or Golda Yelaun for more information. Click here for departmental directory

Main Users of Instrument

1. The main users are General and Honors General Chemistry (Chem 18 and 19). Occasionally other advanced chemistry courses use it
too.

Web Resources
Misc AA300 PerkinElmer info