BIOL 4233 Field Biology
BIOL 5533 Ecological Methods
Lab Protocol Outlines for Experiment 1
LOW-RANGE NITRITE TEST KIT
Hach No. 21820-00
Model No. NI-15
Range: 0-0.5 mg/L
Supplies and reagents
Two color viewing tubes
NitriVer3 reagent powder pillows
Color comparator for NO2
Low-range Nitrite (NO2)
1.
Rinse a color viewing tube several times with the water to be tested. Fill
the tube to the 5-mL mark (first line on the tube).
2.
Use clippers to open one (1) NitriVer3 nitrite reagent powder pillow for
a 5 mL sample. Add the contents of the pillow to the viewing tube. Stopper
the tube and shake for one full minute. [The test results will not be affected
if a small amount of powder does not dissolve.] A pink color will develop if
nitrite is present in the sample. Allow the sample to stand undisturbed for
10 minutes for full color development, but do not wait more than 15 minutes
before proceeding with Steps 3 and 4.
3.
Insert the tube of prepared sample into the right top opening of the color
comparator. Fill the other viewing tube to the 5-mL mark with an untreated
water sample. Insert this tube into the left top opening of the comparator.
4.
Hold the comparator up to a light source, such as a window or the sky or a
lamp, and view through the openings in the front. Rotate the disc to obtain
a color match. Read the mg/L nitrite nitrogen (N) through the scale
window. Results can be expressed as mg/L nitrite (NO2) by multiplying the
final reading by 3.33.
AMMONIA TEST KIT
Hach No. 2241-00
Model NI-8
Range: 0-3 mg/L as ammonia nitrogen
Supplies and reagents
Glass sample tubes
Nessler reagent
Color comparator for NH3
Ammonia (NH3)
1.
Fill the two glass sample tubes to the 5-mL mark (first line on the tubes)
with the water to be tested.
2.
Add three (3) drops of Nessler reagent to one of the tubes, and swirl to
mix.
3.
Allow at least 10, but not more than 25, minutes for color development. If
ammonia nitrogen is present in the sample, a yellow color will develop.
4.
Insert the tube of prepared sample into the right top opening (inside
opening) of the color comparator.
5.
Insert the glass sample tube containing the untreated sample into the left
top opening (outside opening) of the comparator.
5.
Hold the comparator up to a light source such as the sky, a window or lamp
and view through the two openings in the front. Rotate the disc to obtain a
color match. Read the mg/L ammonia nitrogen (N) through the scale
window. Results can be expressed as mg/L ammonia (NH3) by multiplying
the final reading by 1.2.
NITRATE TEST KIT
Hach No. 14161-00
Model NI-14
Range: 0-10 mg/L
Supplies and reagents
Color viewing tubes
Color comparator for NO3
NitraVer6 nitrate reagent powder pillows
NitriVer3 nitrite reagent powder pillows
Low-range Nitrate (NO3) (0-1 mg/L)
1.
Fill one of the color viewing tubes to the lowest mark with the sample to be
tested. Stopper the tube and shake vigorously. Empty the tube and repeat
this procedure.
2.
Fill the color viewing tube to the lowest mark with the sample.
3.
Use clippers to open one NitraVer6 nitrate reagent powder pillow. Add
the contents of the pillow to the sample to be tested. Stopper the tube and
shake for three minutes. Allow the sample to stand undisturbed for an
additional 30 seconds. Unoxidized particles of cadmium metal will remain
in the sample and settle to the bottom of the viewing tube.
4.
Pour the prepared sample into a second color viewing tube carefully so that
the cadmium particles remain in the first tube.
5.
Use clippers to open one NitriVer3 nitrite reagent powder pillow. Add
the contents of the pillow to the sample. Stopper the tube and shake for 30
6.
7.
8.
seconds. A red color will develop if nitrate is present. Allow at least 10
minutes, but not more than 20 minutes, before completing Steps 6-8.
Insert the tube of prepared sample into the right (inside) top opening of the
color comparator.
Rinse the unoxidized cadmium metal from the color viewing tube used in
Step 2. Fill to the mark with the original water sample and place in the left
(outside) top opening of the comparator.
Hold the comparator up to light source such as the sky, a window or lamp
and view through the openings in front. Rotate the disc to obtain a color
match. Read the mg/L nitrate nitrogen through the scale window. To
obtain the results as mg/L nitrate (NO3), multiply the reading on the scale
by 4.4.
High-range Nitrate (NO3) (0-10 mg/L) [run this test ONLY if the first test
yields no results]
1.
Fill one of the color viewing tubes to the mark with deionized water.
Stopper the tube and shake vigorously. Empty the tube and repeat this
procedure.
2.
Rinse the plastic dropper with the sample to be tested, then fill to the 0.5mL mark. Add the contents of the dropper to the rinsed color viewing tube.
3.
Add deionized water to the mark.
4.
Use clippers to open one NitraVer6 nitrate reagent powder pillow. Add the
contents of the pillow to the sample to be tested. Stopper the tube and
shake for three minutes. Allow the sample to stand undisturbed for an
additional 30 seconds. Unoxidized particles of cadmium metal will remain
in the sample and settle to the bottom of the viewing tube.
5.
Pour the prepared sample into a second color viewing tube carefully so that
the cadmium particles remain in the first tube.
6.
Use clippers to open one NitriVer3 nitrite reagent powder pillow. Add the
contents of the pillow to the sample. Stopper the tube and shake for 30
seconds. A red color will develop if nitrate is present. Allow at least 10
minutes, but not more than 20 minutes, before completing Steps 7-9.
7.
Insert the tube of prepared sample into the right (inside) top opening of the
color comparator.
8.
Rinse the unoxidized cadmium metal from the color viewing tube used in
Step 2. Fill to the mark with the original water sample and place in the left
(outside) top opening of the comparator.
9.
Hold the comparator up to light source such as the sky, a window or lamp
and view through the openings in front. Rotate the disc to obtain a color
match. Read the mg/L nitrate nitrogen through the scale window. To
obtain the results as mg/L nitrate (NO3), multiply the reading on the scale
by 4.4.
PHOSPHATE (ORTHOPHOSPHATE) TEST KIT
Hach No. 2250-01
Model No. PO-24
Range: 0-50 mg/L
Supplies and reagents
Square mixing bottle
Two color viewing tubes
PhosVer3 reagent powder pillows
Color comparator for PO4
Low-range Orthophosphate (PO4): 0-1 mg/L
1.
Fill the square mixing bottle to the 20-mL mark with the water to be tested.
2.
Open one (1) PhosVer3 phosphate reagent powder pillow. Add the
contents of the pillow to the mixing bottle and swirl. Allow at least two but
not more than 10 minutes for color development. If phosphate is present, a
blue violet color will develop.
3.
Insert the lengthwise viewing adapter into the color comparator.
4.
Fill one sample viewing tube to about one inch below the top of the tube.
Place this tube into the comparator in the inside tube slot.
5.
Fill the other sample tube with untreated sample water and place into the
comparator in the outside tube slot.
6.
Hold the comparator up to a light source, such as a window or the sky or a
lamp, and view through the openings in the front. Rotate the disc to obtain
a color match. Read the mg/L phosphate through the scale window.
7.
Divide the reading from the scale window by 50 to obtain the mg/L
phosphate (PO4).
Medium-range Orthophosphate (PO4): 0-5 mg/L [Run this test ONLY if the
result of the first test is over-range]
1.
Perform Steps 1 and 2 of the low-range procedure above.
2.
Fill one of the color viewing tubes to the lowest mark with the prepared
sample (approximately 5 mL). Insert this tube into the right top opening
(inside slot) or the color comparator).
3.
Fill the other tube to the lowest mark with untreated sample (approximately
5 mL). Insert this tube into the left top opening (outside slot) of the color
comparator.
4.
Hold the comparator up to a light source, such as a window or the sky or a
lamp, and view through the openings in the front. Rotate the disc to obtain
a color match. Read the mg/L phosphate through the scale window.
5.
Divide the reading from the scale window by 10 to obtain the mg/L
phosphate (PO4).
High-range Orthophosphate (PO4): 0-50 mg/L [Run this test ONLY if the
result of the second test is over-range]
1.
Rinse the square mixing bottle with demineralized water.
2.
Add 2.0 mL of the water to be tested by twice filling the dropper to the 1mL
mark with the sample and discharging the contents into the mixing bottle.
3.
Add demineralized water to the mixing bottle to the 20-mL mark. Swirl to
mix.
4.
Open one (1) PhosVer3 phosphate reagent powder pillow. Add the
contents of the pillow to the mixing bottle and swirl. Allow at least two but
not more than 10 minutes for development of a blue violet color.
5.
Follow Steps 2 and 3 of the medium-range procedure.
6.
Hold the comparator up to a light source, such as a window or the sky or a
lamp, and view through the openings in the front. Rotate the disc to obtain
a color match. Read the mg/L phosphate (PO4) through the scale window.
HACH Company sension5 Conductivity Meter
Conductivity (S/cm or mS/cm)
Total Dissolved Solids (TDS) (mg/L)
Salinity (o/oo)
1.
Press “I/exit” to turn on the meter. (After it auto calibrates, the meter will
display the last value it read.)
2.
Place the probe into the sample water, being careful not to submerge the
taped section.
3.
Press “cond”, then “read”. Wait until the meter beeps and the lock
symbol comes on, then read the sample conductivity. The units will be in
S/cm or mS/cm, depending on the conductivity of the sample. Be sure to
note which units are displayed.
4.
Press “TDS”, then “read”. Wait until the meter beeps and the lock symbol
comes on, then read the sample TDS (total dissolved solids). The units will
be in mg/L.
5.
Press “sal”, then “read”. Wait until the meter beeps and the lock symbol
comes on, then read the sample salinity. The units will be in parts per
thousand (o/oo).
6.
Remove the probe from the sample. Press “I/exit” to turn the meter off.
TURBIDITY
Absorptometric Method
Hach Method 8237
Range: 0-1000 FAU
Reagents and Supplies
DR/890 colorimeter
25-mL sample cells
Procedure
1.
Enter the stored program number for turbidity.
a.
Press PRGM.
b.
Press 95 ENTER.
2.
Fill a sample cell with 10 ml of deionized water (the blank).
3.
Place the blank into the cell holder. Tightly cover the sample cell with the
instrument cap.
4.
Press ZERO. The display will show 0 FAU.
5.
Mix the sample water well. Fill another sample cell with 10 ml of the
sample water.
6.
Place the sample cell into the cell holder. Tightly cover the sample cell with
the instrument cap.
7.
Press READ. The result in Formazin Attenuation Units (FAU) will be
displayed.
APPARENT COLOR
APHA Platinum-Cobalt Standard Method
Hach Method 8025
Range: 0-500 Platinum-Cobalt color units
Reagents and Supplies
DR/890 colorimeter
25-mL sample cells
Procedure
1.
Enter the stored program number for turbidity.
a.
Press PRGM.
b.
Press 19 ENTER.
2.
Fill a sample cell with 10 ml of deionized water (the blank).
3.
Place the blank into the cell holder. Tightly cover the sample cell with the
instrument cap.
4.
Press ZERO. The display will show 0 PtCo.
5.
Mix the sample water well. Fill another sample cell with 10 ml of the
sample water.
6.
Place the sample cell into the cell holder. Tightly cover the sample cell with
the instrument cap.
7.
Press READ. The result in Platinum-Cobalt color units (PtCo) will be
displayed.