EPA-600/4-75-015
November 1975
Environmental Monitoring Series
NATIONAL EUTROPHICATION SURVEY
Data Acquisition and
Laboratory Analysis System for Lake Samples
Environmental Monitoring and Support Laboratory
Office of Research and Development
U.S. Environmental Protection Agency
Las Vegas, Nevada 89114
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RESEARCH REPORTING SERIES
Research reports of the Office of Research and Development,
U.S. Environmental Protection Agency, have been grouped into
five series. These five broad categories were established to
facilitate further development and application of environmental
technology. Elimination of traditional grouping was consciously
planned to foster technology transfer and a maximum interface in
related fields. The five series are:
1. Environmental Health Effects Research
2. Environmental Protection Technology
3. Ecological Research
4. Environmental Monitoring
5. Socioeconomic Environmental Studies
This report has been assigned to the ENVIRONMENTAL MONITORING
series. This series describes research conducted to develop new
or improved methods and instrumentation for the identification
and quantification of environmental pollutants at the lowest
conceivably significant concentrations. It also includes
studies to determine the ambient concentrations of pollutants
in the environment and/or the variance of pollutants as a
function of time or meteorological factors.
This document is available to the public through the National
Technical Information Service, Springfield, Virginia 22161.
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EPA-600/4-75-015
November 1975
NATIONAL EUTROPHICATION SURVEY
Data Acquisition And Laboratory Analysis System For Lake Samples
by
J. W. Mullins, R. N. Snelling, D. D. Moden and R. G. Seals
Monitoring Systems Research and Development Division
Environmental Monitoring and Support Laboratory
Las Vegas, Nevada 89114
ROAP 25ALD
Program Element No. 1BA029
U.S. ENVIRONMENTAL PROTECTION AGENCY
OFFICE OF RESEARCH AND DEVELOPMENT
ENVIRONMENTAL MONITORING AND SUPPORT LABORATORY
LAS VEGAS, NEVADA 89114
-------
DISCLAIMER
This report has been reviewed by the Environmental Monitoring and Support
Laboratory, U.S. Environmental Protection Agency, and approved for publication.
Mention of trade names or commercial products does not constitute endorsement
or recommendation for use.
Effective June 29, 1975, the National Environmental Research Center-Las
Vegas (NERC-LV) was designated the Environmental Monitoring and Support Labo-
ratory-Las Vegas (EMSL-LV). This Laboratory is one of three Environmental
Monitoring and Support Laboratories of the Office of Monitoring and Technical
Support in the U.S. Environmental Protection Agency's Office of Research and
Development.
-------
TABLE OF CONTENTS
Page
LIST OF FIGURES iv
INTRODUCTION 1
GENERAL SYSTEM DESCRIPTION 1
SAMPLE CONTROL 1
LABORATORY ANALYSIS 2
DATA MANAGEMENT 2
APPENDIX A. DATA FORMS 4
APPENDIX B. AUTOANALYZER PROCEDURES 7
TECHNICON TRIPLE CHANNEL AAI'I AUTOANALYZER 7
Reagents for Nitrate Plus Nitrite in Water 9
Reagents for Methyl Orange Alkalinity in Water 12
AUTOMATED PHOSPHORUS ANALYSIS TECHNICON AAII AUTOANALYZER 14
Reagents for Total Phosphorus 15
TOTAL KJELDAHL NITROGEN TECHNICON AAII AUTOANALYZER 18
APPENDIX C. STORET PARAMETER CODES 21
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LIST OF FIGURES
Page
Figure 1. Data acquisition and analysis processing functions 2
Figure 2. National Eutrophication Survey - Data Flow 3
Figure A-l. Field data form 4
Figure A-2. Station description coding record 5
Figure A-3. Sample control card 6
Figure B-l. Nitrate and nitrite in water and waste water 10
Figure B-2. Methyl orange alkalinity 11
Figure B-3. Low level ammonia in fresh and estuarine water 13
Figure B-4. Orthophosphate 17
Figure B-5. Total nitrogen (Kjeldahl) 20
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INTRODUCTION
On December 15, 1971, the U.S. Environmental Protection Agency (EPA) es-
tablished the National Eutrophication Survey (NES) to identify and evaluate
water bodies in the continental United States which have actual or potential
eutrophication problems resulting from phosphorus effluent from municipal sew-
age treatment plants. The specific objective of the survey is to determine if
phosphorus removal from municipal wastes would be expected to benefit the
trophic state of the receiving waters.
Part of the survey consists of characterizing the present trophic state
of the lakes of interest. In situ measurements of depth, temperature, dis-
solved oxygen, conductivity, turbidity, and pH are made at selected points on
each water body. A Secchi disk reading is also obtained.
Discrete samples are obtained at different depths at each sampling sta-
tion. A portion of each sample is filtered in the field, producing a filtered
and an unfiltered portion for each sample. These samples are preserved with
mercuric chloride and shipped to the Environmental Monitoring and Support
Laboratory in Las Vegas, Nevada, (EMSL-LV). There they are analyzed for total
phosphate, dissolved phosphate, ammonia nitrogen, nitrite plus nitrate nitro-
gen, Kjeldahl nitrogen, and alkalinity. A special sample is also obtained
for chlorophyll-a analysis which is performed in the field. Other special sam-
ples are obtained for phytoplankton counts which are performed under contract.
All data are entered in the Water Quality Control Information System (STORET)
for future retrieval and evaluation.
GENERAL SYSTEM DESCRIPTION
Operationally, the processing of samples and data is complex. Several
processes occur simultaneously and are sometimes mutually dependent. Success-
ful processing of samples requires procedures that extend across operational or
organizational functions. Therefore, description of the system is based on
processing functions rather than operational units. The functional operations
are developed as follows:
1. Sample Control Function
2. Laboratory Analysis Function
3. Data Management Function
SAMPLE CONTROL
Sample control personnel record, prepare, and distribute for appropriate
analysis all samples received. Each incoming sample is identified by a unique
laboratory number assigned by the field personnel. Filtered and unfiltered
1
-------
portions are identified by color code. For new sampling stations, location
description information is transcribed from the field data form to a station
description coding record.
Longitude and latitude are determined from U.S. Geological Survey quadran-
gle maps and demographic codes are determined. This coding record is forwarded
for keypunching and entry of a STORET station description record. A four-part
sample control form is generated in Sample Control and accompanies the samples
for analysis. A copy of the field data form, station description coding form
and sample control card is presented in Appendix A.
LABORATORY ANALYSIS
Samples are analyzed using three Technicon Industrial System Autoanalyzers
One system is utilized for Kjeldahl nitrogen on the unfiltered portion of the
sample, one for dissolved and total phosphorus using both portions of the sam-
ple, and one for dissolved ammonia nitrogen, nitrite plus nitrate nitrogen,
and alkalinity. Specific procedures for these analyses are presented in
Appendix B.
DATA MANAGEMENT
The STORET system is utilized for the storage and retrieval of NES data.
No attempt is made here to describe the STORET system. Input format is de-
scribed on the sample control form, station description coding form, and the
field data form. A list of STORET parameter codes is shown in Appendix C as
is a sample of STORET output report. The integration of these elements is
shown in detail in Figure 2.
OUTPUT
Informa-
tion
Reports
Hgure 1. Data acquisition and analysis processing functions
2
-------
iidd Coord., /
Loc. Codes,
etc. /
Figure 2. National Eutrophication Survey - Data Flow
3
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APPENDIX A. DATA FORMS
NATIONAL EUTROPHICATION STUDY
STATION DESCRIPTION (MUST BE COMPLETED FOR NEW STATIONS)
NEW STATION I I
I I I STATE.
4 9
TOTAL DEPTH OF WATER (FT) 1 1 1 j
30
r-ni IMTV
1 AKF MAMF
WORD DESCRIPTION
DATA COOING RECORD
CARD
">
L
O
Zf
T
O
N
'
24 25
V
R
M
O
DY
H
R
M
N
.U.I. ...1.
TOTAL
DEPTH.,
FT
CHLOR-A
y e/L
SECCHI^
IN
1
2
3
4
5
6
7
8
9
10
11
12
13
DEPTH,
FT
X X X X
0
SAMPI.F,
a
xxxxxx
AIR READING
LIGHT,
%
XXX . X
COND ,
pMHO
XXX X
TURB ,
°/oTRANS
xxxx
TEMP ,
"C
X X.X
DO ,
MG/L
XX. X
PH ,
STD
UNITS
X X.X
,
(
,
/
,
t
,
/
,
,
f
(
r
,
,
1
BOD,
4
COMMENTS
ADDITIONAL SAMPLES TAKEN
SEDIMENT CORE | |
PAP 5 GAL CONTAINER I I
OTHER (SPECIFY)
I 1
QUAL CONTROL REPLICATES I |
INITIALS
UMNOl
I 1 TECH
n HELICOPTER
NO
Figure A-l.
Field data form
4
-------
STATION DESCRIPTION CODING RECORD
U.S. ENVIRONMENTAL PROTECTION AGENCY
NERC - LAS VEGAS
NATIONAL EUTROPHICATION STUDY
STORET STATION CODE
1 1 1
4 9
J STATE
COUNTY
CITY
LATITUDE 1 1 1 1 1 1 1 1 i J LONSITUDE 1 L_l
TOTAL DEPTH OF WATER (FT) 1 1 1 i 1 ° 1 MAP
30 80
STATE NAME
LAKE NAME
WORD DESCRPTION
7
10
20
III II III 1
<] »
III II III 1
I
L_I^ L_I_:_
L*-
V
I
n
J
^
1 1 | N| S S
J
=RECISION CODE L—L-ll
ii a
LJJ
10
i i ii i i i i i 4i
.
JO
40
K)
60
70
ty
05
15
25
35
45
55
65
75
85
95
J5
Figure A-2. Station description coding record
-------
1
E i
c
£
o
*
o
o
s
QJ
£
CL Q.
5
SAMPLE IDENTIFICATION
CARD
' DJ
9
>
9
LOCATION
1 1 1
'
,
COLLECTION
YR
MO
1
DY
|
HR
1
MN .
1
,
DEPTH
|
,
SAMPLE NUMBER
ANALYSIS RESULTS
00665
PHOS
\
MG/L
X X
. X
X
X
1 1
f
00671
PHOS -
MG/L
X X . X
Mil
)
X
1
X
,
0061
NH3 -
X
0
N
MG/L
X .
1 1
X X
I
,
00630
NO2+NO3
MG/L
X
X .
1 1
X
1
X
,
00410
TALK
MG/L
XXX
1
,
00625
TKN
MG L
X X . X
1 1 1
,
COMMENTS
SEQ
©M
Figure A-3. Sample control card
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APPENDIX B. AUTOANALYZER PROCEDURES
TECHNICON TRIPLE CHANNEL AAII AUTOANALYZER
General
Channel A -- In the nitrate plus nitrite procedure, the nitrates are reduced to
nitrites using a copper-cadmium reductor column. The nitrites are reacted with
sulfanil amide to form the diazo compound. The diazo compound is coupled under
acidic conditions with N-1-napthylethylenediamine dihydrochloride to form a
reddish purple azo dye. The azo dye intensity, which is proportional to the
nitrate concentration, is then measured.
Channel B -- The method for total alkalinity uses methyl orange as an indicator
because its pH range is in the same range as the equivalence point for total
alkalinity. The color transition is extended by the use of a buffer and
alkalinity is read as being proportional to the loss of color.
Channel C -- The method for ammonia uses the Berthelot reaction in which the
intensity of an indophenol blue color formed by the reaction of ammonia with al-
kaline phenol hypochlorite, is measured. Sodium nitroprusside is used to
intensify the blue color.
Operating Notes
Channel Parameter Range of Operation
A nitrate + nitrite 0.02 - 1.0 mg/liter N
B alkalinity 10 - 200 mg/liter CaC03
C ammonia 0.02 - 1.0 mg/liter N
50 samples/1 hour 3:1 sample/wash ratio
Standards -- Prepare standards fresh when need is indicated;, keep refrigerated
when not in use.
Set 1 - Nitrate plus carbonate plus small amount of ammonia standard
(full-scale standard only) to allow channel C printer to be
started at the proper time
Set 2 - Ammonia
Spiking solution -- The spiking solution will give a theoretical increase of
0.1 mg/liter for nitrate and ammonia and 20 mg/liter for alkalinity when added
at a concentration of 0.1 ml per 10 ml of sample. This solution is prepared
weekly and kept refrigerated.
7
-------
Reagents -- The color reagent for nitrate should be clear and colorless;
refrigerate it. While washing the system, isolate the ammonium chloride rea-
gent line to avoid contamination of ammonia system. Alkalinity requires a
single reagent - methyl orange. On aging, the Brij tends to deteriorate and
must be replenished when debubbling becomes erratic.
Method control measures -- Condition of cadmium reductor column is checked
daily by running a freshly prepared 0.5-mg/liter nitrite standard. Peak height
should be 50% (replace column if peak height exceeds 65% when instrument is
calibrated to read 100% with the regular standard containing 1.0-mg/liter
nitrate.
Condition of ammonia standard is checked daily by running a freshly pre-
pared 0.5-mg/liter ammonia standard. Peak height should be 50% when instrument
is calibrated in the regular manner.
A log is kept of the "Standard Calibration Potentiometer" settings for each
day. When there is a significant change in this setting the cause should be
determined. General trends in this setting are also useful for diagnosis of
problems.
A maintenance log is kept along with the "Standard Calibration" settings
to enable correlation of these two factors. Items to be recorded are reagent
changes, standard changes, lamp replacement, etc.
Quality control measures -- A record is kept on results for replicate and spiked
samples. This record is made available to the Quality Assurance Branch. Every
twentieth sample is run in duplicate and is also spiked and the spike is run in
duplicate.
Routine operation -- The first run of each day consists of a series of ammonia,
nitrate, and alkalinity standards. The 0.5-mg/liter ammonia and 0.5-mg/liter
nitrite standards are also included in this run. The full-scale standards are
repeated at the end of the run. The balance of the tray is filled in with
regular samples.
All subsequent runs begin with a 100% standard for nitrite plus nitrate
and alkalinity followed by a 100% standard for ammonia. The standards are fol-
lowed by two blanks of deionized water, then a series of regular samples. The
run is terminated with both 100% standards and then allowed to print three times
during the final wash to establish a stable baseline. Baseline and standard
calibration settings are not to be changed after the beginning of the run.
Data handling -- Recorder charts for each run are examined for proper peak
shape, washout, and location of printout point. Digital printer tapes are ex-
amined to assure that the proper result is being printed for the corresponding
peak. If standards, replicates, and spikes appear to be satisfactory the re-
sults are transcribed onto sheets to allow results for each individual lake to
be compared.
After all samples for a particular lake are completed and any questionable
results have been rechecked, the results are entered on data cards. The com-
pleted cards are returned to Sample Control for processing before entry into the
STORET system.
-------
General notes -- The cadmium redactor column is removed after flushing follow-
ing the last run and kept inverted in a beaker of water.
The cadmium column is reinstalled after the pump is started and all extran-
eous air bubbles are out of the system. A bypass is installed when the column
is removed.
The 90° heating bath on the ammonia manifold is unplugged after the last
run. Allow warm-up time of 30 minutes before running samples.
Water supply to wash reservoir is shut off after the instrument is shut
down. The Ascarite gas trap is to be kept in place on the water supply vessel
at all times to prevent contamination by ammonia and carbon dioxide.
Sensitivity -- For nitrate plus nitrite the minimum detectable concentration is
0.002 mg/liter. The standard deviation is 0.002 mg/liter below 0.05 mg/liter
and 4% above 0.05 mg/liter.
For alkalinity the minimum detectable concentration is 10 mg/liter. The
standard deviation is 10% from 10 to 50 mg/liter, 5% from 50 to 100 mg/liter,
and 2.5% above 100 mg/liter.
For ammonia the minimum detectable concentration is 0.002 mg/liter. The
standard deviation is 0.002 mg/liter below 0.10 mg/liter and 2% above
0.10 mg/liter.
Reagents For Nitrate Plus Nitrite In Water
Ammonium chloride --
Ammonium chloride (NH^Cl): 10 g
Alkaline water: dilute to 1000 ml
Brij-35* (30% wt/vol solution): 0.5ml
Dissolve 10 g of ammonium chloride in alkaline water and dilute to 1 liter.
Add 0.5 ml of Brij-35 per liter. Alkaline water is prepared by adding sufficient
ammonium hydroxide to distilled water to attain a pH of 8.5.
Color reagent —
Sulfanilamide (C6H8N202S): 20 g
Concentrated phosphoric acid (H3POiJ: 200 ml
N-1-napthylethylenediamine dihydrochloride (C12HlttN2-2HCl): 1.0 g
Distilled, deionized water: dilute to 2000 ml
Brij-35* (30% wt/vol solution): 1.0 ml
To prepare this color reagent, add 200 ml of concentrated phosphoric acid
and 20 g of sulfanilamide to approximately 1500 ml of distilled, deionized
* Manufactured by Technicon Industrial Systems, Tarrytown, New York 10591
9
-------
NITRATE AND NITRITE IN WATER AND WASTE WATER AAIi
(RANGE: 0-1.0 ppm N)
MANIFOLD No. 116-D049-01*
.034 POLYETHYLENE
RECORDER
KASTE
f
I
^^
COLORIMETER
520nm
15mm F/C
1 99 8023 06
2
1
0
•ft, in f/r DUMP TURF
I 5 Turns
A2 i 1700103
REDUCTOR
TUBE
1890000
116 0
) Turns
570370 1]604
JoOefi________
Was
4B9 n BLK/BLK (0.32) AIR
AMMONIUM CHLORIDE
/"«ii itL/YcL (l.ZO)
-~v/ •""" ""
O BLK/BLK (0.32) SAMPLE
1 ^ BLK/BLK (0.32) AIR
89
le
1
J
LUIUK KtAOtRl
^ BLK/BLK (0.32)
•*•
-------
RECORDER
METHYL ORANGE ALKALINITY
(RANGE: 0-200 ppm CaCO3)
MANIFOLD No. 116-D078*
Waste
O
o o
O O
O Ol
LINEARIZER
TO SAMPLER IV
WASH RECEPTACLE
20 Turns
157-0370
0000
To F/C
Pump Tube
COLORIMETER
550nm
15mm * 2.0mm ID F/C
199-B023-06
Waste
GRN/GRN (2.00) WATER
BLK/BLK (0.32) AIR
RED/RED (0.80)
METHYL ORANGE REAGENT
BLK/BLK (0.32) SAMPLE
WHT/WHT (0.60) From F/C
SAMPLER IV
50/Hr
3:1
I Linearizer can be incorporated in this system.
NOTE: Figures in parentheses represent flow rates in ml/min.
TECHNICON INDUSTRIAL SYSTEMS/TARRYTOWN, NEW YORK 10591
Figure B-2. Methyl orange alkalinity
-------
water. Dissolve completely (heat if necessary).
diamine dihydrochloride and dissolve. Dilute to
ionized water. Add 1.0 ml of Brij-35. Place in
refrigerate.
Add 1.0 g N-1-napthylethylene-
2 liters with distilled, de-
brown plastic bottle and
Reagents For Methyl Orange Alkalinity In Water
Buffer, pH 3.1 —
Potassium acid phthalate (C
0.1N_ hydrochloric acid (HC
Distilled, deionized water
3H5KOiJ: 10.2
): 175 ml
Dissolve 10.2 g of potassium acid phthalate in 500 ml of distilled water.
Add 175 ml of 0.IN. hydrochloric acid (8.3 ml hydrochloric acid, sp. gr. 1.19,
per liter of distilled, deionized water) and dilute to 1 liter with distilled,
deionized water. The pH of this solution should be 3.1. If necessary, adjust
with 0.1N_ hydrochloric acid.
Methyl orange (0.05%) --
Methyl orange (C^H^N
Distilled, deionized water
0.5 g
Dissolve 0.5 g methyl orange in 500 ml of distilled, deionized water and
dilute to 1 liter. Filter this solution through glass wool if necessary.
Working buffer indicator --
Methyl orange (0.05%): 85 ml
Buffer, pH 3.1: 250 ml
Brij-35 (30% wt/vol solution)
Distilled, deionized water
0.5 ml
Add 85 ml of 0.5% methyl orange solution to 250 ml of buffer and dilute to
1 liter with distilled, deionized water. Add 0.5 ml Brij-35 per liter.
Ammonia Reagents
Complexing reagent --
Potassium sodium tartrate (KNaC,4Htt06-2H2
Sodium citrate (HOC(COONa)(CH2COONa)2'
Distilled water
0):
2H20):
33 g
24 g
Dissolve 33 g of potassium sodium tartrate and 24 g of sodium citrate in
950 ml of distilled water. Adjust the pH of this solution to 5.0 with concen-
trated suIfuric acid. Dilute to 1 liter with distilled water. Add 0.5 ml of
Brij-35.
12
-------
LOW LEVEL AMMONIA IN FRESH AND ESTUARINE WATER
(RANGE 0-0.5 ppm N)
MANIFOLD No. 116-D082*
ONE CHANNEL
DIGITAL PRINTER
WASTE
RECORDER
COLORIMETER
660nrr
50mm
I.Smir
199 8023-01
50°C
157 B273 32'
TO SAMPLER IV
WASH RECEPTACLE
GRN/GRN (2.00) WATER
20 Turns
157 B089
20 Turns
157 B089
BLK/BLK (0.32) AIR
^
1
IER
nm
nm
mm
01
1 00000 1 00
1 '
^ TO F/C
~~^ PUMP TUBE
F/C
.D.
DO 0000 1 ,
1 1 16 0489
L (
/
/
/
O
RED/RED (0.80) COHPLEXING REAGENT
ORN/ORN (0.42) SAMPLE
o
o
o
WASTE ** O
ORN/ORN (0.42) ALKALINE PHENOL
BLK/BLK (0.32) SODIUM HYPOCHLORITE
ORN/ORN (0.42) SODIUM NITROPRUSSIDE
GRN/GRN (2.00) FROM F/C
iy
3:1
Proportional control needed for healing bath.
NOTE: Figures in parentheses represents (low rales in ml/min.
•TECHNICON INDUSTRIAL SYSTEMS/TARRYTOWN. NEW YORK 10591
Figure B-3. Low level ammonia in fresh and estuarine water
-------
Alkaline phenol --
Phenol (C6H5OH): 83 g
Sodium hydroxide, 20% w/v (NaOH): 180 ml
Distilled water
Using a 1-liter Erlenmeyer flask, dissolve 83 g of phenol in 50 ml of
distilled water. Cautiously add, while cooling under tap water, in small
increments with agitation, 180 ml of 20% NaOH. Dilute to 1 liter with dis-
tilled water.
Sodium hypochlorite (stock) -- Any good commercially available household bleach
having 5.25% available chlorine may be used.
Sodium hypochlorite (working) -- Dilute 200 ml of stock sodium hypochlorite to
1 1iter with water.
Sodium nitroprusside --
Sodium nitroprusside (Na2Fe(CN)5NO-2H20): 0.5 g
Distilled water
AUTOMATED PHOSPHORUS ANALYSIS-TECHNICON AAII AUTOANALYZER
The total phosphorus method utilizes an initial persulfate digestion in a
pressure cooker, followed by the formation and reading of intensity of the
blue-colored complex formed when ascorbic acid reacts with an antimony-phospho-
molybdate complex. The color is proportional to the phosphorus concentration.
Operating Notes
General information -- The manifold is set up for orthophosphate; 0-0.5 mg/liter
phosphorus; 60 samples per hour; 6:1 sample to wash ratio. All samples and
standards are autoclaved with persulfate digestion reagent prior to analysis.
Results are reported as dissolved phosphorus for filtered samples and as total
phosphorus for unfiltered samples.
Standards -- The standards are preserved with mercuric chloride at a concentra-
tion of 40 mg per liter, stored at room temperatures and prepared fresh from
stock solution monthly.
Spiking solution -- The spiking solution is calculated to give an increase of
0.1 mg/liter phosphorus when added at a concentration of 50 A to 10 ml of
sample. This solution is added with a repeating micro!iter dispenser.
Reagents -- The digestion reagent is prepared fresh daily. The molybdate
working reagent is usable for up to 48 hours. The NaCl-Levor IV* (diluent
water) reagent is stable for extended periods.
*Manufactured by Technicon Industrial Systems, Tarrytown, New York 10591
14
-------
Method control and quality control -- A record is kept of the "Standard Cali-
bration Potentiometer" settings for each day. When there is a significant
change in this setting the cause should be determined. General trends in this
setting are also useful for diagnosis of problems.
A maintenance log is kept along with the "Standard Calibration" settings
to enable correlation of these two factors. Items to be recorded are reagent
changes, standard changes, lamp replacement and other routine maintenance.
A record is kept of results for replicate and spiked samples. This
record is made available to the Quality Assurance Branch.
Every twentieth sample is run in duplicate and also spiked, and the spike
run in duplicate.
Standard reference samples of known concentration are occasionally run to
check overall performance of the system.
Routine operation -- The first run of each day begins and ends with a series
of standards and blanks. The balance of the tray is filled with regular
samples.
All subsequent runs begin with a 100% standard and a blank followed with
regular samples. All runs end with a 100% standard and are allowed to print
three times in the final wash to establish a baseline.
Data handling -- Recorder charts are examined for peak shape, washout, and
location of printout point. Printer tapes are examined to assure that the
proper result is being printed for the corresponding peak.
If standards, replicates, and spikes appear to be satisfactory the results
are transcribed onto sheets to allow the results for each lake to be compared.
After all samples for a particular lake are completed and any question-
able results have been rechecked, the results are entered on data cards which
are sent to Sample Control for entry into the STORET system.
Phosphorus digestion procedure -- Ten ml of sample or standard and 0.5 ml of
digestion solution are pipetted into a 16-mm by 125-mm screw cap culture tube.
The samples are placed in an autoclave and digested at 250° F for thirty
minutes. At the end of thirty minutes the autoclave is turned off and allowed
to return to room temperature. The samples at this point are ready for phos-
phorus analysis. The culture tubes are added directly to the sample changer
carousels. The sampling arm is inverted on the sampler to allow clearance.
Reagents For Phosphorus In Hater
Sulfuric acid 3.5 N_ --
Sulfuric acid (h^SOtJ, conc.(sp. gr. 1.84): 179 g
Distilled, deionized water: dilute to 1000 ml
15
-------
Add 179 g of concentrated sulfuric acid to 800 ml of distilled, deionized
water while stirring slowly. Cool the solution and dilute to 1 liter with
distilled, deionized water.
Ammonium molybdate --
Ammonium molybdate ((NHjgMoyO^^O): 27 g
Distilled, deionized water: dilute to 1000 ml
Place 27 g of ammonium molybdate in a glass, 1-liter volumetric flask and
dilute to 1 liter with distilled, deionized water. Visually inspect the solu-
tion for any trace of molybdate's blue coloration. If any blue color is evi-
dent, check the water and glassware for phosphate contamination and make up
new reagent solution.
Ascorbic acid --
Ascorbic acid (C^Og): 18 g
Distilled, deionized water
Dissolve 18 g of ascorbic acid in 800 ml of distilled, deionized water.
Dilute to 1 liter with distilled, deionized water. Refrigerate in brown
plastic reagent bottle.
Antimony potassium tartrate --
Antimony potassium tartrate (KOOCCHOHCHOHCOO(SbO)-%H20): 2.0 g
Distilled, deionized water
Dissolve 2.0 g of antimony potassium tartrate in 800 ml of distilled, de-
ionized water. Dilute to 1 liter with distilled, deionized water.
Diluent water -- Add 5.0 g of sodium chloride to a 1-liter volumetric flask.
Dilute to 1 liter with distilled, deionized water. Add 2.0 ml of Levor IV.
Combined working reagent --
Sulfuric acid (H2S04)5 3.5N; 100ml
Ammonium molybdate ((NHif)6Mo702tt'4H20), 27 g/liter: 30 ml
Ascorbic acid (C6H806), 18 g/liter: 60 ml
Antimony potassium tartrate
(KOOCCHOHCHOHCOO(SbO)-i2H20), 2.0 g/liter: 10 ml
Digestion solution --
Sulfuric acid (H2SOlf), cone., (sp. gr. 1.84): 36.4 g
Ammonium persulfate ((NHtt)2S208): 32.0 g
Distilled, deionized water
Add 36.4 g of concentrated sulfuric acid to 125 ml of distilled, deion-
ized water with stirring. Permit the solution to cool, add 32.0 g of ammonium
persulfate and dilute to 200 ml with distilled, deionized water.
16
-------
ORTHOPHOSPHATE
SAMPLER
1
!^^
i
1 n
u
PC-1
, fc T. f it Wgtu Pi no Tube
5 Turns 37-5° C 20 Turns 20 T
ooo I oooo | oooo oo
7.7ml
-^ To F/C Pump Tube
^
^
•^
WASTE p^ORG/WHT
urns ^^
QO ^YEL/Ytt (1.20) SAMPLE
1
1 f^ BLK/BLK (0.32) AIR
^\J
Q ORG/GRN (0.10) DILUENT WATER
^ BLK/BLK (0.32) WORKING REAGENT
From F/C -. YEL/YEL (1.20)
From F/C p^WASTE RED/RED(0.80)
C
°^/
60/Hr 6:1
^- Debubbler
Figure B-4. Orthophosphate
-------
TOTAL KJELDAHl NITROGEN-TECHNICON AAII AUTOANALYZER
The total Kjeldahl method consists of digesting the organic material with
acid in the continuous digestor, thereby forming ammonium sulfate. The ammonia
is then determined by measuring the color of the indophenol blue complex when
ammonia reacts with alkaline phenol hypochlorite. Sodium nitroprusside is
added to intensify the color which is proportional to the total organic nitro-
gen present in the sample.
Operating Notes
This method involves digestion of the unfiltered sample in a helix type
continuous digestor followed by colorimetric treatment as in the automated
ammonia procedure. The instrument is operated in the range of 0.2 to 5.0
mg/liter nitrogen at 30 samples per hour with a 4:1 sample to wash ratio.
Standards -- The standards are prepared from glutamic acid to be certain the
digestor is operating properly. 500 mg/liter nitrogen stock solution is pre-
pared by dissolving 5.255 g of glutamic acid in water, adding 1 ml of 4% HgCl2
and diluting to 1 liter.
Spiking solution -- The spiking solution is calculated to give an increase of
1.0 mg/liter when added at a concentration of 50 \ to 10 ml of sample. The
spiking solution contains 200 mg/liter nitrogen and is prepared from the
glutamic acid stock solution.
Reagents -- Alkaline phenol: 94 g liquid phenol, 180 ml 20% NaOH, 0.5 ml Brij,
dilute to 1 liter. Refrigerate when not in use.
Sodium nitroprusside: 0.5 g/liter, prepared weekly.
Sodium hypochlorite: 200 ml sodium hypochlorite*/liter.
Sodium hydroxide-tartrate: dissolve 310 g NaOH and 50 g potassium sodium
tartrate in 700 ml water, cool, and dilute to 1 liter.
Digestion reagent: dissolve 3.0 g selenium oxide in 50 ml water, add
20 ml perchloric acid, and dilute slowly to 1 liter with cone. H2S01+.
Startup procedure -- Reagent lines in water, digest line in air; platens on
pumps, pumps off; vacuum pump on, 4 inches of mercury; water aspirator on,
cooling water on; distilled water supply on; bath pumps on, start reagents,
helix rotor on; turn on digestor heaters; invert pulse chamber to fill,
bleed A-4 jacket; check manifold bath temperature 55° C; digestor temperature
- zone 1 360° C, zone 2 320° C; stabilize baseline approximately 20 minutes.
Shutdown procedure -- Heaters off, remove helix cover; reagent tubes in water
except digest and NaOH-tart.; when cooled to 150° C, digest line to air;
continue sodium hydroxide-tartrate solution 5 min., then water 10 min.; pumps
off; break vacuum in waste bottle, vacuum pump off; water aspirator off,
cooling water off; distilled water suppy off; helix rotor off, replace cover.
*Any household bleach will suffice.
18
-------
Emergency shutdown — Remove helix cover; break vacuum in waste bottle; leave
platens clamped on pumps; turn all pump and digester switches off.
Routine operation -- The first run of each day begins and ends with a series
of standards and blanks. The balance of the tray is filled with regular
samples.
All subsequent runs begin with a 100% standard and a blank followed with
regular samples. All runs end with a 100% standard and are allowed to print
three times in the final wash to establish a baseline.
Method control and quality control -- A record is kept of the "Standard Cali-
bration Potentiometer" settings for each day. When there is a significant
change in this setting the cause should be determined. General trends in this
setting are also useful for diagnosis of problems.
r
A maintenance log is kept along with the "Standard Calibration" settings
to enable correlation of these two factors. Items to be recorded are reagent
changes, standard changes, lamp replacement and other routine maintenance.
A record is kept of results for replicate and spiked samples. This record
is made available to the Quality Assurance Branch.
Every twentieth sample is run in duplicate and is also spiked and the
spike run in duplicate.
Data handling — Recorder charts are examined for peak shape, washout, and
location of printout point. Printer tapes are examined to confirm that the
proper result is being printed for the corresponding peak.
If standards, replicates, and spikes appear to be satisfactory, the
results are transcribed onto sheets to allow the results for each lake to
be compared.
After all samples for a particular lake are completed and any question-
able results have been received, the results are entered on data cards which
are sent to Sample Control for entry into the STORET system.
Sensitivity — The minimum sensitivity is 0.2 mg/liter. The standard deviation
is 0.005 mg/liter below 0.5 mg/liter and 10% above 0.5 mg/liter.
19
-------
ro
o
TOTAL NITROGEN (KJELDAHL) - AAII
(RANGE: 0-5 mg/l)
WATER
ASPIRATION
VACUUM PUMP
DIGITAL PRINTER NaOH waste
TO WASTE
OOP
PCI
' A7
•^
?
^ WATER PUR/WHT (3.90)
f XWATER PUR/WHT (3.90) )
X dig/mix PUR (1.70)*
1 Xdig/nm'PUR (1.70)* 1
-~HJ
XX SAMPLE PUR/ORN (3.40)
XN AIR WHT (0.60)
l\J
<-"=»
««
B=tJ
iS»
=«a
SAMPLER IV
-Wft^OAST I Ylf
ii; t err ^«__._
^-^
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O,
1
AIR WHT (0.
PUR/WHT (3
WASTE FROM
PUMP 1
60)
.90) x 4
SAMPLER
TUBES
••M
j
RED (0.80)
^_
-«-KEl-F-TUBING-*- /*
20
QQQO
O
DILUTION WATER PUR/WHT (3.90)
DILUTION WATER PUR/WHT (3.90)
AIR BLK (0.32)
WATER WHT (0.60)
.MIXING
'CHAMBER
HEATING BATH
55°C
IVL_ I oooo I oo oooo oooo tfMHPtr
&*-
COLORIMETER
I 660nm
30mm F/C i
!R
1 I
1
WASTED
^NaOH + TARTRATE RED (0.80)
I X DILUTED SAMPLE RED (0.80)
X ALKALINE PHENOL ORN (0.42)
w
X~\SOD. HYPOCHLORITE BLK (0.3
XNITROPRUSSIDE ORN/WHT (o.^
V-/
LINEARIZER
NOTE: Figures in parentheses represent flow rates in ml/min.
.23)
TO WASTE TRAP
-O
WASTE YEL (1.20)
PUMP II
*ACID FLEX
Figure B-5. Total nitrogen (KJELDAHL)
-------
APPENDIX C. STORE! PARAMETER CODES
LAKE EUTROPHICATION STUDY
STORE! PARAMETER CODES
DATA GENERATED
Field Analysis
"
Field Contact
Sensor
11
"
II
11
"
Field Lab.
NERC-LV
PARAMETER
CODE
00008
00077
00400
00010
00300
00094
00074
00031
32217
00608
00630
00665
00671
00410
00625
COMPUTER
PRINT-OUT
ABBREVIATE
LAB IDENT 'JO.
TRANSP. SECCHI
INCHES
PH SU
WATER TEMP. CENT
DO
MG/L
CNDUCTVY
AT 25° C
MICROMHO
TURB. TRANS. %
INCDT LT
REMNING
PERCENT
CHLRPHYL-A
MG/L
NH3-N DISS
MG/L
N02 & N03
N-TOTAL
MG/L
PHOS-TOT
MG/L P
PHOS-DISS
ORTHO
MG/L
TALK
CAC03
MG/L
TKN
MG/L
DECIMAL
POINT
LOCATION
xxxxxx
xxxxxx
XXXXXX. X
XXXXXX. X
XXXXXX. X
xxxxxx
xxxxxx
xxxxx.x
xxxxx.xx
xxxxx.xx
xxxxx.xx
xxxx.xxx
xxxx.xxx
xxxxxx
xxxxx.xx
PARAMETER DESCRIPTION
Sample number
Transparency, Secchi disk
(inches)
PH (standard units)
Temp., water (° C)
Oxygen, dissolved (mg/1)
Conductivity, Field (micromhos)
Turbidity, transmissometer
(percent transmission)
Light, incident, percent
remaining at certain depth
Chlorophyl-A fluorometric
method (mg/1)
Nitrogen, ammonia
Nitrite + Nitrate
one determ (mg/1 as N)
Phosphorus, total, wet method
(mg/1 as P)
Phosphorus, diss., wet method
(mg/1 as P)
Alkalinity, total (mg/1 as
CaC03)
Kjeldahl nitrogen (mg/1)
GPO 967—356
21
-------
TECHNICAL REPORT DATA
(Please read Instructions on the reverse before completing)
1. REPORT NO.
EPA-600/4-75-015
2.
3. RECIPIENT'S ACCESSION-NO.
4. TITLE AND SUBTITLE
NATIONAL EUTROPHICATION SURVEY
Data Acquisition And Laboratory Analysis System For
Lake Samples
5. REPORT DATE
November 1975
6. PERFORMING 'ORGANIZATION CODE
7. AUTHOR(S)
8. PERFORMING ORGANIZATION REPORT NO.
J. W. Mullins, R. N. Snelling, D. D. Moden and
R- G. Seals
9. PERFORMING ORGANIZATION NAME AND ADDRESS
Environmental Monitoring and Support Laboratory
Office of Research and Development
U.S. Environmental Protection Agency
Las Vegas, Nevada 89114
10. PROGRAM ELEMENT NO.
1BA029
11. CONTRACT/GRANT NO.
12. SPONSORING AGENCY NAME AND ADDRESS
13. TYPE OF REPORT AND PERIOD COVERED
1972-1975
Same as 9.
14. SPONSORING AGENCY CODE
EPA - ORD - Health and
Ecological Effects
15. SUPPLEMENTARY NOTES
Contact J. W. Mullins, (702) 736-2969, extension 326
16. ABSTRACT
A system for data acquisition and laboratory analysis for the National Eutrophication
"Survey is presented. A description is given of the field measurement and data
recording, the sample control process, the laboratory analysis and data management.
Flow charts and data forms are given for the field and sample control functions. A
description with drawings is given of the Technicon Autoanalyzers used for the
laboratory analysis of the samples.
17.
KEY WORDS AND DOCUMENT ANALYSIS
DESCRIPTORS
b.IDENTIFIERS/OPEN ENDED TERMS
COS AT I Field/Group
Data acquisition
Chemical analysis
Water analysis
Lakes
Phosphorus
Nitrogen
Laboratory equipment
Eutrophication
Autoanalyzers
07B
08H
14B
8. DISTRIBUTION STATEMENT
RELEASE TO THE PUBLIC
19. SECURITY CLASS (This Report)
UNCLASSIFIED
21. NO. OF PAGES
26
20. SECURITY CLASS (Thispage)
UNCLASSIFIED
22. PRICE
EPA Form 2220-1 (9-73)
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