“How To”
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Copyright © 2010 SEAL Analytical
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SEAL Analytical - Mequon Technology Center 10520-C Baehr Road Mequon, WI 53092 262-241-7900 www.Seal-us.com
Copyright © 2010 SEAL Analytical
Reproduction or distribution of this document is expressly forbidden without written permission from SEAL Analytical
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SEAL “How To”
Daily Startup
5
Bubble Pattern
6
Wash Solution Baseline Troubleshooting
7
Calculate Reagent Absorbance
8
Calculate Sensitivity
9
Sensitivity / Noise Troubleshooting
10
Colorimeter Check
11
Cadmium Column Preparation
12 - 13
Checking Intersample Air bubble
14
Building a diagnostic package
15
Opening a diagnostic package
16
SEAL Analytical - Mequon Technology Center 10520-C Baehr Road Mequon, WI 53092 262-241-7900 www.Seal-us.com
Copyright © 2010 SEAL Analytical
Reproduction or distribution of this document is expressly forbidden without written permission from SEAL Analytical
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SEAL Analytical - Mequon Technology Center 10520-C Baehr Road Mequon, WI 53092 262-241-7900 www.Seal-us.com
Copyright © 2010 SEAL Analytical
Reproduction or distribution of this document is expressly forbidden without written permission from SEAL Analytical
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SEAL “How To”
CSM-AA3AACE rev 1 01/11
Daily Startup
• Turn on System and Open Software
• Start Charting
HYDRAULIC CHECK – Wash Solution
• Pump system wash solution for 10-15 minutes to allow the system to stabilize.
• Note: Not All Reagent Lines Pump Wash Solution. Check your methods for
exceptions. Typical exceptions: Sampler washpot lines and UV Digester lines
DO NOT contain surfactant.
• Visually check the bubble pattern – refer to Bubble Pattern “How To”
• Bubbles must be the same size, same shape and same distance apart.
• Manifold
• Flowcell
• Waste Lines
• Set Base (Right-click on the chart screen and select “set base”).
• Observe Baseline for several minutes – the baseline should be stable – If
noisy, refer to Wash Solution Baseline “How To”
REAGENT CHECK
• Place reagent lines into the appropriate solutions, pump for 10-15 minutes on
normal speed to allow the system to stabilize.
• Visually check for the correct bubble pattern
• Verify the Gain is set to the normal value for the method.
• Set Base (Right-click on the chart screen and select “set base”).
• Verify that the baseline is stable for several minutes – the baseline should
be stable – refer to Reagent Absorbance “How To”
CHEMISTRY SENSITIVITY
• If you have re-made reagents, stock standards, or changed the pump tubes; the
sensitivity may have changed. Please check your sensitivity – refer to Calculate
Sensitivity “How-To”
• To set Gain, sample the highest calibrant for 2 minutes.
• Double-click on the sampler icon, enter the cup where the high standard is
located, then select “Sample”.
• After two minutes, select “Wash”.
• Primer peak will take between 6-10 minutes to appear.
• When the primer peak reaches plateau, right-click and select “set gain”. This
will adjust the gain so that the peak height will approximately read 90% of the
chart screen.
• Set Base, verify baseline and bubble pattern are stable, and begin analysis.
SEAL Analytical - Mequon Technology Center 10520-C Baehr Road Mequon, WI 53092 262-241-7900 www.Seal-us.com
Copyright © 2010 SEAL Analytical
Reproduction or distribution of this document is expressly forbidden without written permission from SEAL Analytical
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SEAL “How To”
Bubble Pattern
• Bubble shape – Bubbles in Tygon tubing must be round at the front and back. If the
back of the bubble looks straight, the tubing has not be properly wetted and more
surfactant is needed.
Bubble "dragging“ needs
more surfactant
Properly
Wetted
Bubble looks
rounded at front
and back
Bubble looks concave
or square at the back.
Flow direction
• Observe the addition of fluids at each injection point. The addition must be
smooth and continuous with out hesitation or surging.
Too Small
Optimum: length = 1.5 x Diameter
Too Large
If the Bubble Pattern is irregular, check:
• Correct wetting agent is used – per the method
• Correct wetting of the Tygon tubing (from the bubble shape). If the back of the
bubble looks straight, the tubing is not properly wetted and more surfactant is needed
in the system wash solution.
• Correct air injection. Air bubble should be inserted every 2 seconds at each injection
fitting. – Check air valve tubing and function of air valve.
• Regular flow through reagent and sample lines. Irregular flow could indicate a
complete or partial clog in reagent lines / connectors. Replace line and pump tube.
• Correct Pump Tubing and Transmission tubing sizes and connections per method
documentation.
• 116-0528-01 std tubing for lines ≥ 0.60 ml/min
• 116-0536-07 for lines < 0.60 ml/min
• 116-0536-04 for lines ≤ 0.10 ml/min
• Waste line is NOT submerged and drains at bench level
• Waste line length less than 30 cm (12 in)
SEAL Analytical - Mequon Technology Center 10520-C Baehr Road Mequon, WI 53092 262-241-7900 www.Seal-us.com
Copyright © 2010 SEAL Analytical
Reproduction or distribution of this document is expressly forbidden without written permission from SEAL Analytical
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SEAL “How To”
Wash Solution Baseline Troubleshooting
Wash Solution Baseline is unstable or noisy, check:
Regular noise:
• Note the frequency and try to relate to pump components or heat bath
cycling.
Irregular noise:
• Check to see if the flowcell is dirty.
• In AACE 6.04 and higher, the sample : reference ratio on the channel control
window should be between 0.25 and 0.75.
NOTE: If you recently replaced pump tubes, it may take up to 1 hour to achieve a
stable baseline.
Noise peaks – possible causes:
• Electrical. Verify lamp is on – replace if needed.
• Air bubbles. Verify correct bubble pattern into/out of flowcell
Drift:
• With a clean system, the water baseline drift should be less than 1% per hour
at Gain 100 (0.1 AUFS).
• Upward drift - usually indicates a colorimeter fault
• Downward drift - often indicates that a chemical deposit is being washed off
the flowcell. Clean flowcell.
SEAL Analytical - Mequon Technology Center 10520-C Baehr Road Mequon, WI 53092 262-241-7900 www.Seal-us.com
Copyright © 2010 SEAL Analytical
Reproduction or distribution of this document is expressly forbidden without written permission from SEAL Analytical
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SEAL “How To”
Calculate Reagent Absorbance
If the reagent baseline is not stable or is drifting, check the reagent absorbance.
Reagent Absorbance should also be checked when fresh reagents are prepared,
pump tubes were changed or when results are in doubt.
1. Pump wash solution through the system for 5 to 10 minutes.
2. Set Gain to 10. (While charting, double-click on the Channel icon in
the system window and Manually enter the gain as 10.) Do NOT use “Set Gain”.
Note: A Gain setting of 10 sets the full-scale of the chart screen (0% - 100%) to
an absorbance of 1.0.
3. Set baseline (Right-click in the chart window and select “set base”)
• Note the baseline value in chart %. This is your water baseline. (Usually
5.0%)
4. Change over to reagents.
• Once the reagents have reached the detector and the baseline and bubble
pattern have stabilized, note new chart %. This is your reagent baseline.
5. Calculate the reagent absorbance
• Subtract the water baseline percentage from the reagent baseline
percentage and then divide by 100.
Example: water baseline 5%, reagent baseline 15%. 15% - 5% = 10%.
10/100 = 0.10. This is your reagent absorbance (AU).
6. Compare this reagent absorbance to value on method sheet.
• If more than 20% higher, reagents, sampler wash solution or wash pot may
be contaminated.
7. Log the reagent absorbance each time it is checked.
8. Set baseline again before analysis.
Note: Noise-free baseline and low detection limit cannot be achieved when reagent
absorbance is too high.
• For NH3 methods – incorrect pH can cause noise and drifting baseline also. Check
reaction pH in addition to checking reagent absorbance. After any pH adjustment,
recheck reagent absorbance.
• If the baseline was stable with water and drifts or is noisy
with reagents, a common cause is reagent impurity causing
a deposit in the flow cell or a high reagent absorbance.
Reagent
baseline
Water
baseline
Reagent
absorbance
SEAL Analytical - Mequon Technology Center 10520-C Baehr Road Mequon, WI 53092 262-241-7900 www.Seal-us.com
Copyright © 2010 SEAL Analytical
Reproduction or distribution of this document is expressly forbidden without written permission from SEAL Analytical
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SEAL “How To”
Calculate Sensitivity
If the gain has been previously set for an analysis, and your primer is not recognized
by the run, the sensitivity may not be correct and should be checked.
This should also be performed when new reagents are prepared, pump tubes were
changed, or when results are in doubt.
1. Set the colorimeter Gain to 10.
• While charting and with reagents running through the system, double-click
on the Channel icon in the system window. Manually enter the gain as 10.
A Gain of 10 sets full-scale of the chart screen (0% - 100%) to an
absorbance of 1.0. DO NOT USE SET GAIN!
2. Set the base by right-clicking and selecting “set base”.
• Note the baseline value in %.
3. Sample the highest calibration standard for 2 minutes.
• Double-click on the sampler icon, enter the cup where the high standard is
located, then select Sample. When 2 minutes have passed, select Wash
and then close the window.
4. Note the Peak Value in %
• When the peak appears on the chart screen and has a plateau, note the
peak value in %. Then, right click on Charting screen and set Gain.
5. Calculate the sensitivity
• Subtract the reagent baseline percentage from the high standard peak
percentage and divide by 100. See Example calculation below.
• Compare the sensitivity to the value on the method sheet. Target value is
+/-15%.
• If Sensitivity is incorrect or if the plateau is noisy, refer to Sensitivity/Noise
Troubleshooting “How-To”.
Chart %
100 %
= Sensitivity 1.0 at Gain 10
80 %
Sensitivity of
0.75
5%
Example:
• Reagent baseline 5%,
• High standard signal 80%
• 80% - 5% = 75% / 100
= 0.75.
This is your Sensitivity (also
called Absorbance or Extinction)
Baseline
Time
2 min.
SEAL Analytical - Mequon Technology Center 10520-C Baehr Road Mequon, WI 53092 262-241-7900 www.Seal-us.com
Copyright © 2010 SEAL Analytical
Reproduction or distribution of this document is expressly forbidden without written permission from SEAL Analytical
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SEAL “How To”
Sensitivity / Noise Troubleshooting
SENSITIVITY TROUBLESHOOTING
Please Verify:
• Reagents are prepared fresh
• Heating bath temperature is correct and stable.
• For NO3 methods: reduction efficiency is OK (check by running a NO2
standard with the same concentration of N as the high NO3 standard).
• For NH3 methods: pH is in correct range, chlorine reagent is correctly
prepared (strength too high or too low will affect sensitivity) and fresh (solution
should be prepared daily). Stock solution is not stable and should be replaced
monthly.
NOISE TROUBLESHOOTING: the "5-minute check"
• Begin charting with reagents and check that the gain is set at the normal value.
• Set the base by right-clicking and selecting “set base”.
• Note the baseline value in %.
• Sample the highest calibration standard for 5 minutes.
•Double-click on the sampler icon, enter the cup where the high standard is
located, then select Sample. When 5 minutes have passed, select Wash
and then close the window.
• When the signal (peak) appears on the chart screen and has a plateau, check
the steady state signal for noise. If it is noisy, note whether it is random or regular.
Regular noise:
• Measure the frequency and try to relate to pump components or heat
bath cycling.
Irregular noise:
• Check correct bubble pattern and for possible precipitation.
• Check for a dip near the end of the plateau. This effect is usually caused
by the inter-sample air bubble (Inter-Sample Air Compression) – please
refer to the manual to adjust and correct.
Chart %
Steady State
5%
Baseline
5 min.
Time
SEAL Analytical - Mequon Technology Center 10520-C Baehr Road Mequon, WI 53092 262-241-7900 www.Seal-us.com
Copyright © 2010 SEAL Analytical
Reproduction or distribution of this document is expressly forbidden without written permission from SEAL Analytical
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SEAL “How To”
Colorimeter Check
If the baseline is noisy or drifting while pumping system wash solution, please
perform the following check.
1. Switch on all colorimeters and start charting.
2. Fill the flowcell with water with a syringe.
• Disconnect the tubing outside the colorimeter inlet and attach a syringe
filled with DI water with 116-0536-18 tubing.
• Fill the flowcell and leave the syringe attached with the water still in the
flowcell.
3. Perform an Autolamp
• Right-click in the chart screen and select “autolamp”
4. Manually set the Gain to 100 and perform “set base”.
• Wait 5 minutes to allow the colorimeter to stabilize
• Note the chart reading in % and the time on the chart.
5. If, after 30 minutes the baseline is completely noise-free and the drift is less than
1%, then your colorimeter is OK and the source of the problem is probably
hydraulic in nature.
• Bad bubble pattern
• Heating bath
• Dirty flowcell
• Waste line blocked or submerged
6. If the signal is noisy or unstable, an electrical fault may be the source. Write
down the % drift after 30 minutes and colorimeter settings. Please contact
SEAL Analytical for assistance.
SEAL Analytical - Mequon Technology Center 10520-C Baehr Road Mequon, WI 53092 262-241-7900 www.Seal-us.com
Copyright © 2010 SEAL Analytical
Reproduction or distribution of this document is expressly forbidden without written permission from SEAL Analytical
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SEAL “How To”
Cadmium Column Preparation
Step 1: Activate Cadmium
D. Wash the cadmium until
no blue colour remains in
solution.
A. New or used cadmium
particles (10 grams) are
cleaned with 50 mL of 6 N
HCl for one minute.
Rinse the cadmium several
times with distilled water
then decant.
Decant the HCl and wash
the cadmium with another
50 mL of 6 N HCl for one
minute.
E. Add another 50 mL of
2% copper sulfate
pentahydrate and wash
until no blue color remains
in solution or you see
colloidal copper starting to
form.
B. Decant the HCl and
wash the cadmium several
times with distilled water.
Note the presence of small
black copper particles
suspended in the
supernatant liquid.
C. Decant the distilled
water and add 50 mL of
2% copper sulfate
pentahydrate.
F. Decant and wash
thoroughly with DI water
until the solution runs
clear.
In this case, seven
washings were necessary
to remove the suspended
particles.
SEAL Analytical - Mequon Technology Center 10520-C Baehr Road Mequon, WI 53092 262-241-7900 www.Seal-us.com
Copyright © 2010 SEAL Analytical
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SEAL “How To”
Cadmium Column Preparation
Step 2: Packing the Column
A. Close one end of the tygon tubing (18 cm, 116-0536-18) with
glass wool and N6 nipple. (Coarse glass fiber filter paper, nylon
mesh or other inert material may also be used.) To the opposite
end of the nipple, attach additional tubing used for draining.
B. To the other end of the tygon tubing column, attach a
syringe, small funnel, or plastic disposable pipette tip. Attach
this to a clamp on a ring stand (see picture to the left).
C. Using the syringe (or pipette tip) as a funnel, fill the reductor
column with ammonium chloride reagent and clamp the tubing below
the nipple to keep the column filled with ammonium chloride reagent
as shown in the picture to the left.
D. Transfer the prepared
cadmium particles to the
column. Be careful not to
expose the cadmium to air and
do not allow any air bubbles to
be trapped in the column.
E. Pack the cadmium tightly; ensure that no dead
space remains.
F. Close the filling side with glass wool and 2nd N6
nipple. Take free end of drain tubing and connect to
the 2nd N6 nipple and unclamp.
G. After attaching the column to the manifold, but prior to sample
analysis, condition the column. Place a standard of 20 mg N/L Nitrate in
a cup and sample for five minutes. Then do the same with 20 mg N/L
Nitrite for 10 minutes.
H. Total time 20 minutes. It is convenient to prepare a larger amount of
cadmium and prepare several columns at the same time. Sealed as
shown to the left and then stored under buffer solution, they are stable
indefinitely. When they are exhausted, all the cadmium (new and used)
is regenerated together.
SEAL Analytical - Mequon Technology Center 10520-C Baehr Road Mequon, WI 53092 262-241-7900 www.Seal-us.com
Copyright © 2010 SEAL Analytical
Reproduction or distribution of this document is expressly forbidden without written permission from SEAL Analytical
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SEAL “How To”
Checking Intersample Air bubble
Checking for correct operation
While the sampler is sampling normally, watch the inter-sample air bubble as it
passes through the sample tubing. It must be correctly divided at each T-piece.
To perform its function of correctly separating sample and wash, it must
completely fill each sample pump tube as it passes through. When the correct
tubing has been used and the bubble is the right size, the bubble should be
about 3 - 6 mm long in each part of the tubing. Watch to make sure that the
bubble remains in one slug and does not break up in any way.
Troubleshooting
Issue
Solution
Some peaks on some channels look too
narrow and fall off quickly; or poor
1
separation between some peaks: check
point 3.
A. Inter-sample air bubble too small on
channel.
B. Partial clog in sample probe or splitter(s).
2
Inter-sample air bubble is very long, and
breaks up into several smaller bubbles.
Transmission tubing is too narrow.
3
Air bubble fills some sample pump tubes,
but not others.
Stream splitting is not correct: check sample
splitters.
4
Air bubble does not fill any sample pump
tubes.
Inter-sample air bubble is too small.
5
Peaks show spikes at beginning.
Inter-sample air bubble is too large, or
method debubbler not working properly.
SEAL Analytical - Mequon Technology Center 10520-C Baehr Road Mequon, WI 53092 262-241-7900 www.Seal-us.com
Copyright © 2010 SEAL Analytical
Reproduction or distribution of this document is expressly forbidden without written permission from SEAL Analytical
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SEAL “How To”
Building a diagnostic package
If you need to send a diagnostic package to SEAL technical support, please
follow these directions:
•
Open AACE if not already open
•
Select Tools – then Build Diagnostic Package
•
Double click on analysis folder, so that all files open
•
Select the run that needs to be reviewed
•
Click Okay (If using AACE 6.06 see below)*
•
Go to the Start menu
•
Open up My Computer
•
Open up the C: Drive
•
Go to:
Program Files
Seal Analytical
AACE
Data
The Diagnostic package is called: Package_name of run.lzh
•
Attach the Diagnostic package to an email. If sending to SEAL US please
send it to : [email protected]
*Select where you want the file to be saved (AACE 6.06 only), retrieve it
from where it was saved and attach it to an email to: [email protected]
SEAL Analytical - Mequon Technology Center 10520-C Baehr Road Mequon, WI 53092 262-241-7900 www.Seal-us.com
Copyright © 2010 SEAL Analytical
Reproduction or distribution of this document is expressly forbidden without written permission from SEAL Analytical
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SEAL “How To”
Opening a Diagnostic Package
Downloading WinRar or 7-ZIP
•
The diagnostic package file can be opened with the program WinRar or 7ZIP
•
If you do not have either of these programs, go to www.7-zip.org
•
Chose which version is right for your computer and click Download
•
Click Run and then Run again if asked
•
Click Install and then Finished
Opening the diagnostic package
•
Save the attached file to your computer, save it in C: Drive/ Program Files/
Seal Analytical/ AACE/ Data/ Analysis folder
•
You can now go to the file in your C: Drive, Program Files, Seal
Analytical, AACE, Data, Analysis name
•
The diagnostic package will be called: Package_name of run or analysis.lzh
•
Right click on the diagnostic package, Go to 7-zip, then Extract Here
•
Close everything
•
Open AACE
•
All runs should now be accessible
SEAL Analytical - Mequon Technology Center 10520-C Baehr Road Mequon, WI 53092 262-241-7900 www.Seal-us.com
Copyright © 2010 SEAL Analytical
Reproduction or distribution of this document is expressly forbidden without written permission from SEAL Analytical
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