YSI TruLine Ammonia ISE -

Proper Calibration and Measurement Technique

YSI Laboratory Ion Selective Electrodes - Ammonia
Technical Note T623

The measurement of ammonia is critically important in

applications such as wastewater, where treatment plants are
required to provide a laboratory measurement of ammonia
concentration. The ammonia gas-sensing ion selective
electrode (ISE) is an EPA approved method of determining
ammonia concentration for compliance reporting.

AMMONIA ISE MEASUREMENT PRINCIPLE

An ammonia gas-sensing ISE (e.g. YSI TruLine Ammonia
ISE) has two main structures - a pH electrode and a gas
permeable membrane module. A unique ionic strength
adjustor (ISA) is used with the ammonia ISE that buffers the
sample to a pH greater than 11, ultimately causing ammonia
in the sample to become gaseous.
Ammonia is commonly measured in wastewater applications.
The pH electrode fits inside the membrane module and
the sensing end of the electrode is immersed in internal SETTING UP THE AMMONIA ISE
fill solution. Ammonia gas in the sample will pass through
To set up the ammonia ISE, first remove the soaking cap
the permeable membrane, resulting in a pH shift within
from the pH electrode and set aside for use when storing
the internal fill solution. This pH shift is detected by the pH
the electrode long-term. This soaking cap contains pH 4
electrode and can be correlated to the concentration of
buffer and some potassium chloride (KCl).
ammonia in the sample based on the result of calibration.

Carefully fill the membrane module with 15 to 20 drops of

fill solution, then tap the membrane module to ensure no
bubbles are present behind the membrane.

Slide the membrane module over the pH electrode until the

top of the module and the top of the electrode body line up.
Fit the cap over the top of the module and carefully screw
tight. Be careful to not rupture the membrane.

Condition the assembled ammonia ISE by soaking in a

standard solution for at least 15 minutes prior to use. If
completing a two point calibration (e.g. 1 and 10 mg/L) use
the low standard (e.g. 1 mg/L) solution for soaking.

Note: Although not required, it is best to allow the inner pH

electrode to condition within the filled membrane module
for at least 2 hours prior to use. The assembled electrode can
be placed in a low to mid-range standard during this time.
The YSI TruLine Ammonia ISE. A pH electrode is placed inside an outer -continued-
membrane module that is only permeable to ammonia gas.
CALIBRATING THE AMMONIA ISE Step #3: Prepare standards
Determination of effective ion concentration with ion Standards should bracket the expected sample range. For
selective electrodes is very technique sensitive. Although example, if your expected sample range is 5 mg/L, at least
the ammonia ISE is prone to fewer interferences than other one standard must have a lower concentration and one must
ISEs, a great deal of care must be taken when calibrating in have a higher concentration.
order to obtain accurate and repeatable results.
There should at least be a tenfold (i.e. decade) difference in
Step #1: Connect the ammonia ISE concentration between the high and low standards (e.g. 10
Connect the ammonia electrode to a meter that has a BNC mg/L and 100 mg/L, not 10 mg/L and 50 mg/L).
input and can directly display ion concentration (e.g. display
in mg/L), such as the YSI TruLab 1320, MultiLab 4010-2, or At least 2 standards must be used, but the YSI TruLab 1320
the MultiLab 4010-3. Ensure the instrument has been set-up and MultiLab 4010-2/3 can accept up to 7 calibration points.
to measure ammonia.
If your standards span more than one decade (e.g. 1 mg/L
and 100 mg/L), it is best to prepare at least one mid-range
standard (e.g. 10 mg/L).

Each standard should have a volume of 100 mL and should

be placed in a 150 mL glass beaker.

Standards should be fresh and prepared very carefully. It

is best to use a pipette when measuring small volumes of
stock when preparing diluted standards.

Step #4: Place electrode in solution and stir

Place the electrode in the lowest concentration standard
The MultiLab 4010-3 can be used to measure and stir at a constant rate using a stir bar and stir plate.
dissolved oxygen, pH, and ammonia. The stirring speed should be limited to minimize loss of
ammonia gas. Use the same stirring rate when calibrating
Step #2: Connect a temperature sensor and measuring samples.
Connect a temperature sensor to the instrument, as
measuring effective ion concentration is dependent If you do not have a stir plate, it is best to swirl the solution
upon temperature. The standard solutions should have a or use a stir rod once ISA is added.
temperature as close as possible to the expected sample.
Step #5: Add ISA and begin calibration
The YSI ammonia ISE does not have an integrated Add 2 mL of ammonia ISA to the standard. After adding
temperature sensor, but external temperature sensors are ISA, the solution should have a pale blue color, indicating
available (YSI ScienceLine 135 and 136). the pH is > 11.

Alternatively, the “Alternative Temperature” function of the Allow the solution to stir for 1 minute and begin calibration.
TruLab and MultiLab can be used to apply a temperature Calibration is time sensitive, as ammonia in the sample is no
reading from another sensor that is connected to the longer sufficient ~4 minutes after ISA is added. Therefore,
instrument (e.g. use the temperature reading from a MultiLab ensure calibration with the standard is complete 1 to 4
IDS pH sensor). minutes after ISA is added. This timing is critical! If calibration
using the standard is unsuccessful within this time limit, a
fresh standard will need to be prepared.

Note: The MultiLab and TruLab have an Auto-Read feature

that is used during calibration. If the instrument has not
YSI ScienceLine Temperature Sensors determined the reading is stable, but you are confident the
reading is stable, the <Enter> key can be pressed to accept
the point and move on to the next.

T623 | Technical Note | Ammonia Laboratory ISE

 If the electrode will not calibrate, you have ensured the
electrode is correctly assembled, and you are confident
your procedure is correct, attempt to clean the pH electrode
and/or change the membrane module (see Maintenance
section).

Step #8: Recalibrate often

The ammonia electrode should be calibrated at the
beginning of each day.

Verify your calibration result every 2 hours by preparing a

fresh low to mid-range standard, adding ISA, and verifying
the reading. If the mV reading has changed ~3 mV compared
to the reading in that standard during calibration, you will
need to recalibrate the electrode.

TAKING A MEASUREMENT
It is critically important that samples be prepared using the
same procedure used for standards. Therefore, follow steps
#3-5 of the calibration procedure when preparing samples
and taking a measurement.

In summary, 100 mL of each sample should be collected and

2 mL of ISA should be added. The sample should be fresh
and the same stirring rate used during calibration should be
Ideal set-up for calibration. A stir plate, 150 mL glass beaker, temperature utilized. A measurement can be determined 1 to 4 minutes
sensor (e.g. from pH electrode), ISE instrument, and ISA are all used. after ISA is added. The Auto-Read function of the TruLab and
MultiLab can be used to ensure the measurement is stable.
Step #6: Calibrate with additional standards
Once the instrument has accepted the first calibration point, TROUBLESHOOTING
finish calibrating using steps #4 and #5 for the remaining The following tips may prove useful if you are experiencing
calibration points. Make sure to calibrate in order of issues with the ammonia ISE such as unacceptable
increasing concentration. calibration results or poor electrode response.

If calibrating with less than 7 standards, you can finish Refresh the internal fill solution
calibration by pressing the <M> key after calibrating with If the internal fill solution does not shift back to its original
your highest concentration standard. A complete calibration pH before another sample is taken, the next pH shift is
record will be displayed. not completely attributed to the new sample or standard.
Therefore, if you are having issues with poor slope or
If calibrating with 7 standards, the TruLab and MultiLab will drifting readings, the electrode fill solution may need to
display a complete calibration record after calibrating with be refreshed.
the last standard.
To ensure the internal fill solution pH shifts back to its original
Step #7: Evaluate electrode slope pH, you may need to:
After calibrating, evaluate the electrode slope on the
calibration record. For highest accuracy, the ammonia 1. Manually refresh the fill solution that is between the tip of
electrode slope should be between -53 mV/decade and the pH electrode and the membrane. This can be done by
-65 mV/decade. holding the electrode body sensor side down, pulling the
cable back and away from the body/membrane slightly (it
If the electrode slope is out of this range, attempt to is spring loaded to cause the glass to seat up to the inside
recalibrate. Ensure your standards have carefully been of the membrane) and then slowly releasing the cable. This
prepared, ISA was used, and calibration to each point was pulls the glass away from the membrane, causing the internal
completed 1 to 4 minutes after ISA was added. fill solution to refresh quickly in the interface area.
 ELECTRODE STORAGE
Properly storing the ammonia ISE will help ensure good
electrode response and repeatable results.

The internal fill solution between the membrane and the tip of the pH sensor Short-term storage
can be refreshed by slightly pulling the cable away from the assembled Between measurements, the ammonia ISE should be stored
electrode. A spring in the electrode cap allows for this. in a low concentration standard, such as 1 or 10 mg/L, with
2. Allow more time between measurements for the pH of ISA added.
the internal fill solution to return to normal.
For overnight or weekend storage, place the assembled
3. Shaking out the old, and refilling the membrane with electrode in 1000 mg/L standard and do not add any ISA.
new internal fill solution every few days might also be useful.
These short-term storage conditions will keep the assembled
Check for a tear in the membrane electrode stable for a comparably long period of time. Do
To check for a tear in the membrane, place the fully not remove the membrane module for short term storage,
assembled electrode in pH 4 buffer. Since the membrane as this can stretch the membrane, ultimately causing the
is only gas permeable, there should be no aqueous ion electrode to be less responsive.
transfer between the inside and outside of the membrane.
Therefore, if the mV reading on the instrument display Long-term storage
changes drastically once the electrode is placed in pH 4 For long-term storage, disassemble the electrode and rinse
buffer, there is a potential leak or tear in the membrane the module and pH electrode with DI water. Carefully dry
module. all parts of the electrode. Place the pH electrode inside the
included soaker bottle with pH 4 buffer and KCl added as a
MAINTENANCE storage solution. The membrane module can be stored dry.
If the electrode has a poor response, you may attempt to
clean the pH sensor much like other pH electrodes. Avoid COMMON QUESTIONS - FAQs
rubbing or scratching the glass sensor.
#1: My electrode from another manufacturer was
It may also prove useful to condition the pH electrode by shipped dry. Is this OK?
placing it in pH 4 buffer with some KCl added. Ensure the YSI never recommends storing the electrode dry. In fact, no
reference (i.e. the white layer where the glass changes in pH electrode should ever be stored dry!
dimension) is completely immersed.
The storage solution the TruLine Ammonia ISE is shipped in
The membrane module will also need to be changed if it contains the proper balance of moisture and salts in order
is excessively dirty or stretched. These modules can last a to prevent leaching of reference electrolyte and the active
long time in clean samples. However, in dirty samples like ingredients of the glass membrane. If you are storing the
wastewater where NH3 measurements are common, they electrode long-term, be sure to place the pH electrode
may have a much shorter life. inside the included soaker bottle with pH 4 buffer and KCl
added as a storage solution. If you do not have KCl, pH 4
Membrane modules need to be replaced when the buffer will suffice.
electrode no longer calibrates or responds. The module will
eventually develop pinholes or tears (likely from mashing #2: Do I need to recalibrate after using the spring-loaded
the membrane into solids at the bottom of the beaker!), get cable to refresh the fill solution?
stretched, or the membrane will become fouled (surface
coating). The internal fill solution (IFS) is homogenous, but an
air bubble or other issues may change the tiny volume
You may attempt to clean a membrane module that has between the membrane and the tip of the pH electrode (i.e.
surface contamination by soaking it in DI water for 1 hour or the interface where the pH shift occurs). If the user is very
more, then soak in a low or mid-range standard for at least consistent in their preparation (e.g. consistent amount of
15 minutes prior to use. IFS in the interface) and use of the electrode, recalibration
is not needed. However, we recommend recalibration if the
user is relatively new to the ammonia ISE.
#3: If measuring at low concentrations, is it OK to wait #5: How often should I replace the membrane and/or
longer than 4 minutes after adding ISA to calibrate or the internal fill solution (IFS)?
measure?
There is no general guideline for this, as the replacement
It is not possible to generate more ammonia gas than what interval simply depends how often the ammonia ISE is used,
is in solution, so there is no reason to wait longer. Ammonia the samples measured (are they dirty?!), and the storage
will still become gaseous at the same rate regardless of practices used. Some customers that use the ammonia
concentration. ISE every day choose to replace the IFS once a week and
the membrane once a month, while other customers only
If the user waits longer than 4 minutes, ammonia will start change the membrane and IFS when needed.
to be depleted and the calibration/measurement result will
be negatively impacted. If experiencing issues, YSI recommends recalibrating first.
If that does not resolve the issue, change the IFS. If issues
If you are measuring in low level samples and having persist, change the membrane. The internal pH electrode
difficulty, consider using multiple samples in low level should be the last component replaced.
measurement applications for highest accuracy. In addition,
some customers have had success using Standard Method Indications that a membrane or IFS change is needed
4500-NH3 E (i.e. the known addition method) rather than include a slow electrode response, the calibration slope
Standard Method 4500-NH3 D (i.e. the direct calibration is outside the recommended range, and/or the cable pull
method) when calibrating. technique no longer helps.

They key to success with any gas-sensing ammonia electrode #6: How often should I replace the internal electrode?
is to have a good, consistent technique!
The internal pH electrode is just like any other pH electrode,
so you should expect a similar usable life. Also like other pH
#4: Is it best to insert the ammonia ISE at an angle so air
electrodes, proper storage of the pH electrode is critical!
bubbles don’t get trapped on the membrane?
A pH electrode should never be stored dry or in deionized
The consistent presence of a bubble on the surface of the (DI) water.
membrane is not a concern, as ammonia gas can easily
permeate the bubble and then the membrane. In addition, Customers that use the electrode extensively should not
inserting the ammonia ISE into the solution at an angle is have to replace their electrode any more than once a year.
not feasible on many electrode holders. Some customers report getting at least two years of life out
of their electrode. In order to maintain consistency, some
Although the consistent presence of a bubble is not a customers choose to replace their electrode once a year.
concern, bubbles sporadically moving across the membrane
should be prevented. This sporadic movement of bubbles
across the surface of the membrane should not occur as Chris Cushman is an Assistant Product Manager at YSI, a
long as the membrane is all the way in solution and the stir Xylem brand and can be reached at christopher.cushman@
plate speed is not turned up too high. xyleminc.com or at 937.767.7241, x279

YSI Inc. | 1725 Brannum Ln | Yellow Springs, OH 45387

Tel +1.937.767.7241 | 800.897.4151 | [email protected] | @YSIinc

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