This booklet provides step-by-step detailed instructions for the Code 3633-05 test
kits. It is important to review these instructions thoroughly
perform the tests by the short-form instructions contained in the case lid.
To order individual reagents or test kit components, use the specifi ed code number.
before attempting to
Testing Hints / Reagent Care ........................................................................ 4
1. Clean glassware is a must for accurate results.
Thoroughly rinse test tubes before and after
each test. Caps and stoppers should also be
cleaned after each use.
2. Use test tube caps, not your fi ngers, to cover test
tubes and fl asks during shaking or mixing.
3. When adding sample to calibrated test tube, be
sure vial is fi lled to the appropriate mark. The
bottom of the liquid (meniscus) should be level
with the desired mark. (Fig. 1)
4. When dispensing reagents from bottles fi lled
with dropper plug and cap, be sure to hold
bottle vertically and gently squeeze to dispense
the appropriate number of uniform drops.
(Fig. 2)
5. For those reagents to be added with the screwcap
pipet assemblies
on bottle and replace with the screwcap pipet.
NOTE: Place the polyseal caps back on the
reagent bottles for longer periods of storage. Be
sure that both pipet assemblies and polyseal caps
are thoroughly cleaned before placing on bottles
to avoid contamination.
6. When dispensing reagents from pipets, hold
pipet vertically to assure uniform drop size. This
is extremely important when performing drop
count titrations. (Fig. 3)
7. To fi ll pipets, squeeze rubber bulb and immerse
into reagent. Release bulb to fi ll. (Fig. 4)
8. To accurately dispense powdered reagents with
spoon, tap spoon on edge of reagent container
to remove excess reagent. (Fig. 5)
9. When performing tests that include Octa-Slide
Comparators, the comparator should be
positioned between the operator and non-direct
sunlight. This allows the light to enter through
the
This test kit uses two basic analytical procedures common to fi eld test kits. A brief
explanation of each follows:
COLORIMETRIC: OCTA-SLIDE 2 VIEWER
In a visual colorimetric test, a sample is treated with reagent(s) to produce a color
reaction, generally in proportion to the amount of test factor present. The sample color
is then compared against color standards representing known concentrations of the
factor being tested over a specifi c range.
1
123
4Note:
4
Insert OctaSlide 2
into the OctaSlide 2
(1101).
Match sample color
to color standard.
Record results.
Bar
Viewer
23
Insert test tube
containing reacted
sample into the
Octa-Slide 2
Viewer (1101).
Hold the Octa-Slide 2
Viewer so that nondirect light enters
through the
back of
the viewer.
Note:
If sample color is between two standards, the midpoint
is taken as the result.
If the sample is darker than the highest standard, a
dilution may be performed on a fresh sample, and the
test repeated to bring the concentration within range.
DILUTIONS
The calibrated test tubes (0106) included in this kit may be used to perform dilutions
for the Ammonia Nitrogen and Nitrite Nitrogen tests. Distilled or deionized water is
needed to perform dilutions.
The following table provides a quick reference guide for dilutions of various
proportions. Once the dilution is prepared, use this diluted sample to perform the test,
and multiply the result by the dilution factor to obtain the actual concentration.
Sample SizeDistilled Water to Bring to 10 mLDilution Factor
In a titrimetric method, titrating solution (or titrant) is added to a treated sample
until a color change occurs. The volume of titrant required to reach this endpoint is
proportional to the concentration of the factor being tested. Direct Reading Titrators
provide results directly in the appropriate concentration for the test – no counting of
drops, no calculations.
1.0
0.9
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
PlungerBarrelAdapter Tip
The Titrator consists of a plastic barrel, a plastic plunger, and a plastic adapter tip. The
adapter tip reduces the size of the drops that are dispensed, increasing the precision of
the test results. DO NOT remove the plunger or adapter tip from the Titrator.
1
12 3
Fill the test
tube to the
appropriate
line with
sample water.
45
To fi ll Titrator, slowly
4
withdraw the plunger
until the bottom of the
plunger is
opposite
the zero
mark on
the scale.
7
6
Insert the Titrator
into the center hole
of the test tube
cap. While gently
swirling tube, add
titrating solution
one drop at a time
until the desired
color changes occur.
Follow individual
test instructions.
23
Add reagents
as specifi ed
in the instructions
for the individual
test method. Cap
with the special test
tube cap. Mix by
swirling gently.
If small air
appear in
them by
the barrel
the
titration solution back
into
the reagent container.
bubbles
the barrel, expel
partially fi lling
and pumping
Depress the
Titrator plunger
to expel air. Insert
Titrator into the
plastic fi tting of
the titrating
solution bottle
and invert.
6
Turn the bottle
5
right-side-up
and remove the
Titrator.
Repeat until bubble
disappears.
89
7
8
When testing is
complete, discard
titrating solution in
Read the test
result directly
from the scale
where the large
ring on the Titrator
meets the Titrator barrel.
Titrator. Do not return
the titrant to the
reagent bottle. Rinse
Titrator and titration
tube thoroughly. Do
not remove the plunger
or the adapter tip from
the Titrator.
Proper control of water quality is an essential part of successful aquaculture operation.
Immediate test results provided by on-site water analysis equipment can confi rm a healthy
environment, or give early warning signals for required treatment.
1. Develop a routine testing schedule.
2. Keep records! Historical data is extremely important if treatments are required. Note
environmental conditions, fi sh activity, feeding habits, etc.
3. Observe fi sh to note any particular behavior or feeding rates, as this may be a sign
of stress.
4. Stable characteristics, such as alkalinity and hardness, do not have to be tested as
frequently as ones that fl uctuate, such as ammonia nitrogen, nitrite nitrogen, pH,
dissolved oxygen and temperature. Keep in mind that these factors fl uctuate throughout
the day and in some cases are interdependent.
5. Be alert to sudden changes in one factor, as it may be a clue to perform further analysis.
Test Tube, 5-10-12.9-15-20-25 mL, glass, w/cap0608
Direct Reading Titrator, 0-200 Range0382
*WARNING: Reagents marked with an * are considered to be potential health hazards.
See page 6 for more information.
The Direct Reading Titrator is calibrated in terms of total alkalinity expressed as parts
per million (ppm) Calcium Carbonate (CaCO3). Each minor division on the Titrator
scale equals 4 ppm CaCO3.
ALKALINITY TEST PROCEDURE
1
1
Fill the test
tube (0608)
to the 5 mL
line with the
sample water.
4
45
Fill Direct
Reading
Titrator
(0382) with
*Alkalinity
Titration
Reagent B
(4493DR).
7
7
Read the test result
directly from the
scale where the large
ring on the Titrator
meets the Titrator barrel.
Record Total Alkalinity as ppm
Calcium Carbonate (CaCO3).
23
23
Add 4 drops
of BCG-MR
Indicator
(2311-PG).
5
Insert the
Titrator into
the center
hole of the
test tube cap.
8
Note:
If the plunger tip reaches the bottom
line on the scale (200 ppm) before the
endpoint color change occurs, refi ll
the Titrator and continue the titration.
When recording the test result, be sure
to include the value of the original
amount of reagent dispensed (200 ppm).
Cap and
mix.
Solution
will turn
blue-green.
6
6
While gently
swirling the tube,
slowly press the
plunger to titrate
until blue-green
color changes
to pink.
Ammonia in water occurs in two forms: toxic unionized ammonia (NH3) and the
relatively non-toxic form, ammonium ion (NH
forms as ammonia-nitrogen (NH3-N) to give the total ammonia-nitrogen concentration
in water. The actual proportion of each compound depends on temperature, salinity,
and pH. A greater concentration of unionized ammonia is present when the pH value
and salinity increase.
1. Consult the table below to fi nd the percentage that corresponds to the temperature,
pH and salinity of the sample.
2. To express the test result as ppm Unionized Ammonia Nitrogen (NH3-N), multiply
the total ammonia-nitrogen test result by the percentage from the table.
3. To express the test result as ppm Ionized Ammonia Nitrogen (NH4-N), subtract the
unionized ammonia nitrogen, determined in Step 2, from the total ammonia-nitrogen.
Percentage of Free Ammonia as (NH3) in Freshwater1 (FW)
and Seawater2 (SW) at varying pH and temperature.
10°C15°C20°C25°C
pHFW1SW2FWSWFWSWFWSW
7.00.190.270.400.55
7.10.230.340.500.70
7.20.290.430.630.88
7.30.370.540.791.10
7.40.470.680.991.38
7.50.590.4590.850.6651.240.9631.731.39
7.60.740.5771.070.8361.561.212.171.75
7.70.920.7261.351.051.961.522.722.19
7.81.160.9121.691.322.451.903.392.74
7.91.461.152.121.663.062.394.243.43
8.01.831.442.652.073.832.985.284.28
8.12.291.803.322.604.773.736.555.32
8.22.862.264.143.255.944.658.116.61
8.33.582.835.164.067.365.7810.008.18
8.44.463.546.415.059.097.1712.2710.10
8.55.554.417.986.2811.188.8714.9712.40
1
Freshwater data from Trussel (1972).
2
Seawater values from Bower and Bidwell (1978). Salinity for Seawater values = 34 ppt at an
ionic strength of 0.701 m.
FOR EXAMPLE:
A fresh water sample at 20°C has a pH of 8.5 and the test result is 1.0 ppm as total
Ammonia-Nitrogen.
1. The percentage from the table is 11.18% (or 0.1118).
2. 1 ppm total Ammonia-Nitrogen x 0.1118 = 0.1118 ppm Unionized AmmoniaNitrogen
Test Tube, 5-10-12.9-15-20-25 mL, glass, w/cap0608
*WARNING: Reagents marked with an * are considered to be potential health hazards.
See page 6 for more information.
CARBON DIOXIDE TEST PROCEDURE
1
1
Fill the test tube
(0608) to the
20 mL line
with the
sample water.
3
45
Fill Direct Reading
Titrator (0380) with
Carbon Dioxide
Reagent B
(4253DR).
6
7
Read the test result
directly from the
scale where the large
ring
meets the Titrator
barrel. Record as
Carbon Dioxide.
water
sample
on the Titrator
23
Note:
For best results, test
a freshly obtained
sample, and avoid
splashing or prolonged
contact with air.
4
Insert the
Titrator into
the center
hole of the
test tube cap.
Note:
Result:
40 ppm
The Titrator is calibrated in terms
of carbon dioxide expressed as
ppm Free CO2. Each minor division
on the Titrator scale equals 1.0
ppm CO2.
2
Add 2 drops of
*Phenolphthalein
Indicator, 1%
(2246). If sample
turns red, no free
carbon dioxide is
present. If sample
is cololess,
proceed to Step 3.
5
6
While gently
swirling the
tube, slowly
press the
plunger to
titrate until a
faint pink color
develops and
persists for 30
seconds.
Test Tube, 5-10-12.9-15-20-25 mL, glass, w/cap0608
*WARNING: Reagents marked with an * are considered to be potential health hazards.
See page 6 for more information.
The Titrator is calibrated in terms of chloride expressed as ppm Cl_. Each minor division
on the Titrator scale equals 4.0 ppm Cl_.
HIGH CHLORIDE AND SALINITY READINGS
For high chloride and salinity readings the sample must be carefully diluted to bring it
within a practical range for titration. Dilutions of 1 to 20 or 1 to 100 are recommended.
(For example: 1 mL of sample water is diluted to a total of 20 mL with distilled water.
This is a 1 to 20 dilution.) Titration tube is then fi lled to 15 mL line with diluted
sample, and the titration is performed as described. The Titrator reading is multiplied
by the appropriate conversion factor given below for parts per million (ppm), parts per
thousand (ppt), or percent (%) Chloride.
1 to 20 DILUTION
ppm chloride=Titrator Reading x 20
ppt chloride=Titrator Reading x 0.02
% chloride=Titrator Reading x 0.002
1 to 100 DILUTION
ppm chloride=Titrator Reading x 100
ppt chloride=Titrator Reading x 0.1
% chloride=Titrator Reading x 0.01
To convert parts per thousand (ppt) Chloride to parts per thousand (ppt) Salinity use
the following formula:
Fill the test tube
(0608) to the
15 mL line
with the
sample water.
4
Cap and swirl
to mix.
Solution will
turn yellow.
78
While gently
swirling the
tube, slowly
press the
plunger to
titrate until
yellow color
changes from
yellow to orange
or orange-red.
Add one drop of
*Phenolphthalein
Indicator, 1%
(2246). If sample
turns pink, add
*Sulfuric Acid,
0.5N (6090) one
drop at a time,
mixing after each
drop, until pink
color disappears.
5
Fill Direct
Reading
Titrator (0382)
with *Chloride
Reagent #2
(4505DR).
Read the test result directly
from the
ring
Titrator barrel. Record as ppm
Chloride.
scale where the large
on the Titrator meets the
Add 3 drops
of *Cloride
Reagent #1
(4504).
6
Insert the
Titrator into
the center
hole of the
test tube cap.
Note:
If the plunger tip reaches the bottom line on the scale (200 ppm) before
the endpoint color change occurs, refi ll the Titrator and continue the
titration. When recording the test results be sure to include the value of the
original amount
Test Tube, 5-10-12.9-15-20-25 mL, glass, w/cap0608
Pipet, plain, plastic, w/cap0392
Water Sampling Bottle, 60 mL, glass0688-DO
*WARNING: Reagents marked with an * are considered to be potential health hazards.
See page 6 for more information.
The Titrator is calibrated in terms of Dissolved Oxygen expressed as ppm Dissolved
Oxygen. Each minor division on the Titrator scale equals 0.2 ppm Dissolved Oxygen.
NOTE: Be careful not to introduce air into the sample while adding
the reagents.
1
Remove the
cap from the
bottle.
3
Cap the bottle and mix by
inverting several times.
A precipitate will form.
5
Add 8 drops of
*Sulfuric Acid, 1:1
(6141WT).
2
Immediately add 8
drops of *Manganous
Sulfate Solution (4167)
and add 8 drops of
*Alkaline Potassium
Iodide Azide (7166).
4
Allow the precipitate
to settle below the
shoulder of the bottle.
6
Cap and gently invert the bottle
to mix the contents until the
precipitate and the reagent have
totally dissolved. The solution
will be clear
yellow to
orange if the
sample contains
dissolved oxygen.
NOTE: At this point the sample has been “fi xed” and contact between the
sample and the atmosphere will not aff ect the test result. Samples may be
held at this point and titrated later.
Fill the titration tube
(0608) to the 20 mL line
with the fi xed sample.
Cap the tube.
3
Insert the Titrator into
the plug in the top of
the Sodium Thiosulfate,
0.025N (4169) titrating
solution.
0
.
1
0
0
.
2
.
3
0
0
.
4
5
0
.
0
.
6
.
7
0
8
0
.
.
9
0
0
1
.
2
0
0
0.1
0.1
Depress plunger of
0.2
0.2
0.3
0.3
0.4
0.4
the Titrator (0377).
0.5
0.5
0.6
0.6
0.7
0.7
0.8
0.8
0.9
0.9
1.0
1.0
4
Invert the bottle and
slowly withdraw the
plunger until the large
ring on the plunger is
opposite the zero (0)
line on the scale.
NOTE: If small air bubbles appear in the titrator barrel, expel them by
partially fi lling the barrel and pumping the titration solution back into the
reagent container. Repeat until bubble disappears.
1.0
0.9
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
20
5
Turn the bottle
upright and remove
the Titrator.
Note:
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
If the sample is a
very pale yellow,
go to Step 9.
Insert the tip of
the Titraror into
the opening of the
titration tube cap.
7
Slowly depress the
plunger to dispense the
titrating solution until
the yellow-brown color
changes to a very pale
yellow. Gently swirl the
tube during the titration
to mix the contents.
8
Carefully remove
the Titrator and
cap. Do not disturb
the Titrator plunger.
10
Cap the titration
tube. Insert the tip
of the Titrator into
the opening of the
titration tube cap.
Read the test result directly from the scale
12
where the large ring on the Titrator meets
the Titrator barrel. Record as ppm Dissolved
Oxygen. Each minor division on the Titrator
scale equals 0.2 ppm.
9
Add 8 drops of Starch
Indicator Solution
(4170WT). The sample
should turn blue.
11
Continue titrating until the blue
color disappears and the solution
becomes colorless.
NOTE: If the plunger ring
reaches the bottom line on
the scale (10 ppm) before the
endpoint color change occurs,
refi ll the Titrator and continue
the titration. Include the value
of the original
reagent dispensed
when recording the test result.
amount of
(10 ppm)
NOTE: When testing is complete, discard the
titrating solution in the Titrator. Rinse Titrator
and titration tube thoroughly. DO NOT remove
plunger or adapter tip.
*WARNING: Reagents marked with an * are considered to be potential health hazards.
See page 6 for more information.
The Titrator is calibrated in terms of Total Hardness expressed as parts per million (ppm)
Calcium Carbonate (CaCO3). Each minor division on the Titrator scale equals
4 ppm CaCO3.
ANALYSIS OF HARDNESS IN SALT WATER
When sea and estuarine waters containing very high levels of mineral salts are to be tested,
the sample must be diluted before titration. This test set is supplied with a calibrated pipet
for perforning the simple, convenient dilution described below:
1
Use the 0.5 mL
pipet (0353) to
add 0.5 mL of the
salt water to the
test tube (0608).
2
Fill the test tube
to the 12.9 mL
line with distilled
water (a 1 to
25.8 dilution).
3
Follow Steps 2 through
9 under Hardness Test
procedure. Multiply
reading by 25.8. Record
as ppm Total Hardness
CaCO3.