O&A HYDROCHEMISTRY
Document:
WI_Nut_007
Version:
1.1
Date:
04/05/2017
AA3 Protocol for Running Samples
AA3 protocol for running samples
Start of voyage on the processing computer
 Create Folder in Hydrochem directory V:current\hydrochem\in2015_v01_AA3_files
Start of voyage on the AA3 Acquisition computer
 Select Configure, then “configure software”

Choose V:\current\hydrochem\in2015_v01_AA3_files
This will ensure a copy is saved to the current directory for the processing computers. The
files are also stored on the acquisition computer.
1
Also ensure this other settings are correct:
2
3
4
5
6
Setting up Master analysis and folder for Voyage
Select setup analysis
Select New analysis
Select OK
7
Type in voyage name, this becomes the
folder name. e.g.in2015_v01
Add in description e.g. 5
nutrients
Enter sample and wash
time; 80:40
8
Select
units;
µmol/L
Select
Slope only
Enter in
concentration of
stds and select 4
decimal places.
Type in each nutrient analysis name under each
channel, exactly like this; 1=SILICATE,
2=PHOSPHATE, 3=NITRITE, 4=NOx, 5=AMMONIA.
Otherwise Hypro will not recognise.
Select Peak Parameters
9
Under Peak
parameters,
Select Before primer
rise acceptance
If a noisy channel, e.g.NO2
increase the noise tolerance.
10
Ensure there is not a .
after the Cal – Hypro
will not recognise.
Select Tray Protocol. Can either
type this in or select “Load Tray”
and load an existing tray setup.
When entering or deleting, make
sure “Fix” is selected.
11
Enter Drift
Sample
Check before
drift
Enter LNSW
before
baseline
Enter Baseline
Before End. Hypro
uses this Baseline.
When finished tray protocol & all
other pages, select OK
Before starting analysis you must do the following, select the sample tray configuration. Usually
24 for the first tray and 60 for the second tray. Also make sure you use “Calibrants above 900” is
selected. If using “Custom Rack” make sure to select.
Also select wash pump ON.
Select OK on all pages as you close them down.
12
13
To begin analysis
1. Switch on everything before selecting
2. Move air-line tubing either back or forwards
3. Rinse and fill MQ H2O container.
Select charting
4. Clamp down platen and peristaltic pump on sampler.
5. Let MQ H2O run through system for at least ½ hour
6. “Set Light Power” lamp should be 11.89 V, right hand click on mouse.
7. Set baseline at 5%
14
8. Switch in reagents and let run for at least 15 minutes before starting analysis (Note: if running
NO3 let NH4Cl buffer run for a couple of minutes before switching in Cd column). Make sure
baseline is stable.
9. Note % reagent baseline, then reset to 5% - record in AA3 Run analysis sheet.
10. Select “Set Up” and then “Analysis/Run” – double click.
11. Add in sample ID’s into tray protocol.
Note: Ensure names in Tray Protocol are in conventional naming format for hypro. For instance Do
not use full stops (beware AACE adds a full stop after Cal-please remove this), ASW, or #. Any
unusual names add TEST in front of the name, e.g. Test12345 or TestSpecialSample.
12. Change the filename to the following format 150302A-in2015_v01nut001. The 001 is
consecutive number and is the run number.
13. If you are using a new Cd column activate it before starting the analysis using 250 ul of stock
NO3 std in 30 ml LNSW.
14. Set Gain by sampling highest Cals, if running SiO4 140 µmol/L then set Silicate with Cal 6.
15
Note: setting the gain is only required the first time the analysis is run using the master
program. After this the gain settings will always be the same. It is a good idea to still check
them. That the high cal is still 90% . If the gain is different manually set the gain back to 10
which sets the AUFS to 1.000, then run the high cals through and click on set gain.
14. Set Gain by sampling highest Cals, if running SiO4 140 µmol/L then set Silicate with Cal 6.
15. Wait for baseline to stabilise and return to 5%, if not reset.
16. Visually check bubble pattern is good, then you are ready to start.
Select
Stop
Select Run & choose the run file
you have just made e.g.
150302A-in2015_v01nut001
17. Watch to ensure the baseline is found, if not found after significant amount of time then click
on the
force baseline icon.
18. Ensure the Primer is found. If it is not found stop analysis.
19. At the end of run watch to see if final baseline is found, if not then press
the force
baseline.
20. Switch off Cd column and then all reagents to water, rinse for 5 minutes then place all
reagents straws into 10% Hypochlorite solution allow this to run for 10 minutes and then place all
reagents straw back into MQ H2O and rinse for 30 minutes.
21. Unclamp platen and remove. Unclamp peristaltic pump on sampler. Loosen pump tubing on
manifold.
16
All analyses are run without smoothing, baseline, carry-over & drift corrections. These are
performed in Hypro.
The AA3 is now run without having the “Auto stop and Save if Primer not found” box checked
within the Configure Software page. As this can cause the run to stop prematurely.
on manifold.
Processing the analysis within SEAL
Go to Retrieve and
select view chart
Select the file you just
ran.

All Processing now occurs in Hypro.

However just check over trace within AACE to see if there are any really bad peaks that
may need #.
17
Press the
to export the file, choose ASCII file.
Under Format heading ensure that “Sylk” format is selected.
18

Under Export Data ensure the following boxes are checked;
Click Export then Save &
Close
 The exported files will have the following format;
150329A_in2015_v01nut001.sylk and 150329A_in2015_v01nut001.chd
 They will be found in the following directory
C:\Program Files\SEAL Analytical\AACE 6.10\Data
 Copy the files into the following directory V:current\hydrochem\hypro\Nutrients


Rename the files by removing the 150329A_, so that they have the following filenames;
in2015_v01nut001.sylk and in2015_v01nut001.chd (std format inYYYY_v##nut###).
Files can now be processed in Hypro.
Before processing sylk and chd files within Hypro fill out the AA3 Processing worksheet and #
any bad peaks (place # in front of number in the peak start column), remove any full stops,
ensure conventional naming format for sample ID’s or put Test in front of any unconventional
names all within the sylk file. Hypro also now allows you to change peaks within the Analysis
Trace to BAD (operator) they will turn red.
19

Settings for windows in Hypro; Phosphate 50-100%, Nitrate 65-105% and Silicate 50-105%.
However you will select the green “peak window” by clicking on the start and end position
you want.
Peak Window


Check all data and record all QC information in the AA3 processing work sheet.
Side Note: If Want to view AACE Files on Processing Computer and create backup on
network
On the Processing computer go to Program Files and Select SEAL6.10, paste a copy of the
folder “in2015_v01” which contains the Master file (anl) into the Data folder.

The folder “in2015_v01” is copied from the C:\Program Files\SEAL Analytical\AACE
6.10\Data on the aquistion computer.

Go to V:current\hydrochem\in2015_v01_AA3_files

Select the files from the run you just analysed, there is usually 10 files that look like the
following;
20

Copy these files into C:\Program files\Seal Analytical\AACE 6.10\Data\in2015_v01
21
Version information
Version # Date
IDENT Changes made
Status
IDENT
Approved
sch531
Approved
she384
Click for date Ident
Status
Ident
Click for date Ident
Status
Ident
Click for date Ident
Status
Ident
Click for date Ident
Status
Ident
Click for date Ident
Status
Ident
01.0
20/10/2015
ree13f
Document created
01.1
5/05/2017
she384 Addition of version control and theme
22