A STUDY OF PRECISION AND ACCURACY OF
LABORATORIES. AND METHODS OF ANALYSIS
OF AMMONIA NITROGEN IN LAKE MICHIGAN WATERS
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TABLE OF CONTENTS
Page
LIST OF TABLES iii
LIST OF FIGURES iv
I. INTRODUCTION 1
II. PROCEDURES. CHRONOLOGICAL DEVELOPMENT " 2
III. FINDINGS AND DISCUSSION 7
17. CONCLUSIONS AND REGOMMFNDATIONS 10
BIBLIOGRAPHY 18
APPENDIX . • 19
Procedural Details 20
Key to Symbols and Abbreviations in the Tabulated Data 25
ii
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LIST OF TABLES
No. Page
la, Ib Ammonia Nitrogen Determinations by Distillation 26
Average Results (mg/l)
2 Ammonia Nitrogen Determinations by AutoAnalyzer 28
Average Results (mg/l)
3 Average Recoveries of Ammonia Nitrogen by Distillation 29
and AutoAnalyzer (mg/l)
lia, Ijb, Ammonia Nitrogen Determinations by Distillation 30
Uc, Ud Original Data (mg/l)
5 Ammonia Nitrogen Determinations by AutoAnalyzer 3h
Original Data (mg/l)
6 Ammonia Nitrogen Recoveries by Distillation and
Auto Analyzer, Original Data (mg/l) 35
iii
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LIST OF FIGURES
No. Title Page
1 Percent Distribution of all Ammonia Nitrogen
Determinations, 0.00 and 0.02 mg/1 11
2 Percent Distribution of all Ammonia Nitrogen
Determinations, O.OU, 0.05, and 0.06 mg/1 12
3 Percent Distribution of all Ammonia Nitrogen
Determinations, 0.11, 0.12, and 0,15 mg/1 13
U Percent Distribution of all Ammonia Nitrogen
Determinations, 0.30 and 0.38 mg/1 lU
5 Percent Distribution of all Ammonia Nitrogen
Determinations, 0.72, 0.75, and 0.97 mg/1 IS
6 Ammonia Nitrogen Recoveries from Spiked Lake Samples
by Distillation and AutoAnalyzer (0.5 and 0.10 ng/1) 16
7 Ammonia Nitrogen Recoveries fron River Sanples
by Distillation and AutoAnalyzer (1.0 mg/1) 17
8 AutoAnalyzer Methodology. Amrtonia Nitrogen Flow Diagram 2U
9 Probability Curve. Ammonia Nitrogen Determination
by Distillation - O.OU mg/1 Standard Sample, April 26,
1966 - 36
10 Probability Curve. Ammonia Nitrogen Determination
by AutoAnalyzer - O.Oh mg/1 Standard Sample, April 26,
1966 ' 37
11 Probability Curve. Recovery of Ammonia Nitrogen,
Determination by Distillation - 0.05 mg/1, April 26,
1966 38
12 Probability Curve. Recoverr of Ammonia Nitrogen,
Determination by AutoAnalvzer - 0.05 i^g/l, April 26,
1966 39
13 Probability Curve. Recovery of Ammonia Nitrogen,
Determination by Distillation - 1.0 mg/1, March 23,
1966 liO
lU Probability Curve. Recovery of Ammonia Nitrogen,
Determination by AutoAnalyzer - 1.0 mg/1, March 23,
1966 111
iv
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A STUDY OF PRECISION AND ACCURACY OF LABORATORIES AND
METHODS OF ANALYSIS OF AMMONIA NITROGEN IN LAKE MICHIGAN WATERS
A Report to the Technical Committee of the Calumet Area-
Lake Michigan Enforcement Conference
by
The Laboratory Directors of the Calumet Area-Lake Michigan
Enforcement Conference
I. INTRODUCTION
This is a report of comparison studies performed by cooperating
laboratories concerned with analysis of Lake Michigan waters and stream
waters in the Calumet Area of Indiana and Illinois.
A series of comparison studies is to be made for determinations of
ammonia nitrogen, phosphate, phenol, cyanide, and threshold odor. Part I
presents the findings from a series of five comparison studies on ammonia
nitrogen completed between September 9, 1965 and April 26, 1966.
•
The objectives of the comparison studies are:
1, To determine the reliability (precision and accuracy) of the
analytical procedures as normally used in the desired concentra-
tion range.
2. To determine if there is a change in the accuracy of these
procedures arising from departures from prescribed analytical
routine.
3. To determine the degree of variation between the participating
laboratories.
U. To find the sensitivity of the methods as used.
5. To recommend necessary changes in procedures.
The laboratories participating in the ammonia nitrogen study were:
1. American Oil Company
2. Cities Service Oil Company
3. City of Chicago, Bureau of Water, Dept. of Water and Sewers
li. Garv-Kobart Water Corporation
5. Great Lakes-Illinois River Basins Project
6, Indiana State Board of Health
7. Inland Steel Company
8. Lake Huron Program Office, GLIRBP
• 9. Lake Ontario Program Office, GLIRBP
10. Metropolitan Sanitary District of Greater Chicago
11o Robert A. Taft Sanitary Engineering Center
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II. PROCEDURES. CHRONOLOGICAL DEVELOPMENT
This section of our report presents a brief chronological commentary
on the conditions, implied and explicit, which obtained in the various
Test Series. It is intended to provide background for assessment of the
results obtained in individual Series and also for comparison cf any or
all Series. All results, excepting those of Series I, are summarized in
Tables 1 to 6 in the Appendix.
SERIES I. September 9, 1965
Number of samples analyzed: Four: Raw Lake T-,'ater, preserved and
unpreserved, from Gary-Hobart and from Chicago South District
Filtration Plant.
Conditions: Samples stored at room temperature 2lt hours,
Participating laboratories:
1. Citv of Chicago, Pureau of '.rater, Dept. cf "rater and Sewers
2. Gary-Hobart "ater Corporation
3. Great Lakes-Illimis P-ivar Pas ins Project
lu Metropolitan Sanitary District cf Greater Chicago
There was no
As it turned out, of the four participating
same method. The results, as reported, were widely divergent. Average
values from the four participating laboratories are shown below in mg/1:
Chicago— SD?p Gary—He tart
Unpreserved
0.021
0.160
0.05
0.01
Preserved
0.082
C.200
0.09
0.02
-npreservea
O.C26
0.150
°.C6
OoCl
Preserved
0.110
0.260
O.C6
0.02
These results revealed that the laboratories were consistent -.;ithin
their own methods, but no two laboratories agreed as to the concentration
of ammonia found.
Because of the wide disagreement, the Technical Committee, Calumet
Area-Lake Michigan "nfcrcenent Conference, requester" th"t comr^rison ~LuJies
be made bv laboratories concerned with Calumet Area surveillance. The
GLIRBP liberator*- prepared a working draft cf the proposed comparison studies
These were distributed to the Laboratory Directors for review.
September 30, 196g
A meeting of the Calumet Area Laboratory Directors, under the chair-
manship of r.T, Clifford Paisley, Jr0j) was held to discuss and revise the
proposed procedures for future comparison studies. The -.embers cf the
Technical Committee of the Calumet Area-Lake Michigan Fjiforcement Conference
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met with the Laboratory Directors during the first hour to review the
problem and. offer their suggestions.
The Laboratcr7 Directors agreed that Series II should consist of
amonia nitrogen, phosphate, phenols, cyanide, and odor standards and
samples.
SERIFS II. October 13, 1965
Number of sarples for ammonia nitrogen: Nine (5 synthetic)
(h raw water)
Additionally: phosphates - 10 samples
phenols -5 "
cyanide - 8 "
odor -5 "
Participating laboratories:
1. American Oil Co.
2. City of Chicago, bureau of "later, Dept. of "later and Sevrers
3. C-ary-Mcbart "Tatar Corporation
li. Great Lakes-Illinois River Basins Project
5. Indiana State Board of Health
6. Inland Steel Cor.oany
/ • -'
Analysis date: raw water 10/13/65; synthetic lC/lh/65.
*
Conditions: Collection, preservation, and. distribution procedures
;re standardised and time for analyses S'Tichrcnized. Porr.s "era c'is-
'_-^u_5i, ~ cr r~"cr^_n.^ r£~u._-s Ln "ornTj_2.o^."uej 'T-9re\'^r "or.ic z.ca j_e»
to be duly reccixled on th -r
Conditions ':ere stioulated for Distillation ~rccedure a3 f.:llovc:
with emphasis on the following points;
"1. "he distillation apparatus should be stear.ied out by nlaoing 5CC i-l
of distilled rater and 1C ml of phosphate buffer solution in the flaskj
stean out un~il the distillate is free of ammonia.
2. Preserved sanples and standards should be adjusted to a r" of
seven, using a oH r.eter orior to adding the buffer solution.
3. -Tse a 5CO ml sample or standard and distill over aprroxinately
250 ml, collecting the distillate in 00002^ sulfuric acid. The distillate
is then concentrated by evaporation on a hot plate. The resulting solution
should be mads uo to 100 ml and nesslerized. This gives a concentration
factor of 5. Standards h and 5 need not be concentrated b^ evaporation.
L. The method use:' to read the samole vrill be indicated on the report.'
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The Sodium Phenate method was also outlined for the Technicon
AutoAnalyzer.(2) However, the final paragraph of these instructions
read as follows: "Any method or methods that are being used by any of
the laboratories may be employed in these analyses. The method used
should be spelled out for the purpose of interpreting the results."
At the November 29 meeting of the Laboratory Directors, the follow-
ing points were made in discussing the results of the last comparison,
Series II. Considerable variation existed among the laboratories both in
their procedure and in the results reported. Each laboratory, however,
displayed good reproducibility within itself. Values obtained by the
Technicon method were judged best, followed in order by distillation and
direct nesslerization. The following excerpts from the November 29 meeting
are pertinent at this point:
"1. It appears that some of the laboratories were not experienced at
running analyses in the low range (0.00-0,06 mg/l) desired and/or
by the prescribed methods. It also appears that too much pres-
sure was placed on the laboratories to accomplish a large amount
of work in a short period of tine. It was recommended that the
study be conducted again, at a more relaxed pace, and with a
fixed number of samples and standards for each run.
2. An indispensable condition for conducting a successful inter-
laboratory "study is skill and familiarity with the method on the
part of all participants. The methods used, when applied to the
low concentrations found in lake water, i.e. at or near the
maximum sensitivity of the method, cannot be used successfully
without a great deal of skill and familiarity with the method on
the part of the individual analyst. This was not the case in
this series of comparison studies.
3. Identical copies of a detailed description of all steps to be
followed in this test procedure for comparison studies should
be provided to each participant.
ii. One standard sample, plus a sample of Gary raw water and
Chicago raw water, is enough for each laboratory- to check at
one time.
5. Although one laboratory used research qualitv instrumentation,
the Spectronic 20 is considered to be good enough to give the
desired results."
Under the chairmanship of Mr. LeRoy Scarce, the Laboratory Directors
met February 17, 1966. Here it was decided that future studies would be
limited to one parameter at a time. Ammonia nitrogen was selected for the
next test series, using standard samples only,,
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SERIES III. March It, 1966
Number of samples analyzed: Six (synthetic)
Participating laboratories:
!• American Cil Company
2. Cities Service Cil Company
3. City of Chicago, Bureau of Water, Dept. of Water and Sewers
U. Garv-Hobart Water Corporation
5. Great Lakes-Illinois River Basins Project
6. Indiana State Board of Health
7. Inland Steel Company
8. Lake Huron Program Office, GLIHBP
9. Metropolitan Sanitary District of Greater Chicago
Analysis date: March I;, 1966
Conditions: The following cciditicns were stipulated bv advance
notice tc the participating laboratories.
w
Distillation Ilethod: Standard Methods, 12th Ed., pp 186-193, with
emphasis on the follo-:ing points:
''Steaming cut apparatus to ensure amr.onia-free conditions -
same as in previous series.
Adjustment to pH 6.6 -crior to adring buffer - previous .series
called for pH 7.0.
Collection of distillate in 0.02!' sulfuric acid - previous
series called for C.00211 sulfuric acid.
Concentration of distillate to 50 ml instead cf ICO ml - as
required in previous series."
Procedure for AutoAr.alyzer remair.ed unchanged frc.~ last Series.
March 1?, 1966
Calumet Area Labcratorv Directors met for the purpose cf reviewing
the results obtained from the Karch It Test Series III. The results from
Series III showed a substantial improvement over Series II. From a review
of these results, it was agreed t,hat an additional study should be made;
it was requested and agreed that a prescribed procedure for the ammonia
nitrogen test be provided for use in future comparison tests. This was
done as requested, with particular emphasis on distillation of standards
in preparing calibration curve and volume c^ distillate„
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SERIES IV. March 23, 1966
Number of samples: Six
Participating laboratories:
1. American Oil Company
2. Cities cervice Oil Company
3. City of Chicago, Bureau of Water, r
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At the meeting of the Calunet Area Laboratory Directors May 19, 1966,
for the purpose of reviewing recent comparison Series V for ammonia nitrogen,
the Laboratory Directors unanimously decided that further testing would be
of little value. Series V showed that when all laboratories followed the
same procedure, most of the laboratories obtained an accuracy of +_ 0.02
mg/1 or better, in analyzing standard samples containing 0.00 and O.Oii mg/1
ammonia nitrogen.
The subject of direct nesslerization was again brought up and the
opinion was expressed that this method should not be entirely excluded from
the future thinking of the members of this group.
Problems which existed throughout the Series were as follows:
From the beginning of the Series there was a question of ammonia-free
water and ammonia-free reagents (buffer and sulfuric acid). Ordinary
distilled water was found to contain significant amounts of ammonia.
Redistilled water, upon concentration, still contained some ammonia. The
best procedure for obtaining armionia-free water was found to be the addition
of sulfuric acid to the redistillation, if a carbon or ion-exchange column
couldn't be attached to the still. Once the ammonia-free water was prepared,
keeping it free from ammonia contamination was an important factor.
Clean glassware was imperative. Cleaning glassware with chromic acid
solution and then rinsing with ammonia-free water was found to be the best
preparation for glassware.
It was suggested that rubber stoppers be replaced with.special neoprene
stoppers since ammonia does adsorb to rubber.
This section on procedures indicates that with conformity of procedure,
reliable results can be obtained.
The results of the above comparison series are discussed and summarized
in detail in the next section of this report.
III. FINDINGS AND DISCUSSION
Distillation Method
As indicated in the preceding section, progressive improvement in
agreement of results from the individual laboratories was realized in
application of the distillation method. Uniform test procedures were not
followed by all laboratories for the first three series and results were
erratic„ After uniform procedures were followed, the results were more
consistent and gave better indications of the accuracy that can be achieved
when the recommended procedure is carefully followed.
Tables la and Ib present individual laboratory average results
chronologically and anonymously. When analyzing samples containing 0.00 mg/1
of amnonia nitrogen, an accuracy of + O.OU mg/1 (one standard deviation) was
obtained in Series II, whereas in a repeat analysis in the last series, an
accuracy of + 0.02 mg/1 was realized. The same improvement was noted when
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analyzing standard samples containing concentrations in the range of 0.02
to 0.05 mg/1. In general, the average values indicate a slight tendency
to be on the high side in these determinations.
Even closer agreement was demonstrated when analysing Lake Michigan
samples from the City of Chicago and Gary-Kobart water intakes, where a
precision of +_ O.OU mg/1 was obtained in October 1965 (Series II), and in
April 1966 the" precision had improved to _+ 0.01 mg/1.
Recoveries of known amounts of ammonia nitrogen by the distillation
process are shown in Table 3. The precision and accuracy were, in
this instance, the same and was + 0.02 mg/1 for recoveries of 0.10 and
0.0^ mg/1 amr.onia from Lake Michigan water. The precision and accuracy for
recovery of 1.0 mg/1 ammonia nitrogen from river water (collected from the
Indiana Harbor Canal) was +0.29 rag/1.
AutcAnalyzer Method
From the inception of the studies, the AutoAnalyzer method provided
results demonstrating precision and accuracy superior to those cf the
distillation method. These results are shown in Tables 2, 5, and 6.
Precision and accuracv, ir terms of one standard deviation for standard
and lake samples,' was most often + 0.01 mg/!0"or the recoveries of the
known amounts 'C.Cp, 0.10, and 1.0 ng/l) added to lake and river water,
the accuracy was +_ 0.01, + 0.01, and +_ 0.09 mg/1, respectively.
Data Evaluation
Figures 1 through 7 summarize the results from both methods in terms
of deviation from the known amounts of ammonia nitrogen. The range of
deviation from known amount and distribution cf individual values are
presented as percent of all determinations accruing, from all participating
laboratories. The reliabilitv of the method as. used in the various
laboratories mav, in this manner, be estimated. Approximately 32 to 96'-?
of all the distillation determinations for standard samples ranging from
0.00-0.06 mg/1 are within +_ 0.02 mg/1 (one1 standard deviation). It is to
be noted that a significant percentage of AutoAnalvner determin?ticno shewed
no deviation at all from the known amounts '(60 to 90^).
Forty-nine to 5?3 of all distillation results were "ithin +_ C.02 mg/1
for standard samples in the range 0.11-C.l!? mg/1 ammonia nitrogen; 22-SI/'
were within +_ O.OU for standard samples in the range 0030-0.38 mg/1 ammonia
nitrogen; and u5-6l^ were within +_ 0008 mg/1 for standard sarrples contain-
ing 0.?2-0o97 mg/1 ammonia nitrogen.
The findings are presented in further graphic detail in Figures 9
through III, located in the Appendix in the form of probability curves fcr
each sample in the entire study.
Laboratory Evaluation
Since the data are herein presented anonymously, a direct comparison
of laboratory performance in using the distillation method is net possible.
However, all laboratories, of course, did not show equal performance. As
indicated in the Procedure Section, consistency of relative achievement was
present at the beginning of these comparisons. Most of the laboratories
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did demonstrate^that, with enough effort and special precautions, the
data so generated might be considered as interchangeable if a standard
deviation of +_ 0.02 mg/1 is allowable.
A statement by Mr. Dwight G. Ballinger, consultant to the Laboratory
Directors, is quoted below:
"Adequate data is lacking on the precision and accuracy of the
ammonia nitrogen determination at concentrations below 0.1 mg/1.
Standard Methods, Twelfth ^dition (196;?), indicates a repro-
ducibility of 5% for the distillation procedure, but this
figure was obtained at concentrations found in polluted waters.
In a recent study, Dr. David .Jenkins, University of California,
investigated ammonia nitrogen concentrations of approximately
0.03 mg/1 in San Francisco Bay waters. Dr. Jenkins found that
the standard deviation of the distillation-Nessler procedure,
using 10 replicate determinations of the same sample, was
+_ 0,005 mg/1. Experience in the Analytical Reference Service
of the Taft Center indicates that a comparison of results
between laboratories yields a deviation at least three times
greater than the deviation within the single laboratory. Thus,
an extrapolation of the Jenkins data suggests an interlaborator-r
comparison of approximately +_ 0.02 mg/1 at the 0.03 mg/1 level.
It should be noted that the University of California studies
were conducted with 10 replicate determinations, in a very
carefully controlled test. The results obtained are probably
superior to those normally found in routine survey ",:crk.
On the basis of the available information on the precision snd
accuracy of the ammonia nitrogen ceterriination, it is mv
opinion t:~a~ the values rfnortec bv the average laboratcrv may
be expected to deviate from the true concert ration ''.- a^ least
+ C.02 mg/1 at ammonia nitrogen levels below C.C5 mg/1. How-
ever, in order to ensure the best possible results, only tho:?e
laboratories demonstrating abilitv to meet this precision level
should be included in a nutrient monitoring operation."
In conclusion, this section on procedure shows that good results car. be
attained in the distillation method for ammonia nitrogen, especially at the
low levels encountered in Lake Michigan water, provided that: a great amount
of care in preparation and skill in technique are exercised, and that there
is strict adherence to the method outlined in Standard Methods with the
modifications as described in the Appendix of this report.
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10
IV. CONCLUSIONS-AND RECOMMENDATIONS
1« This study indicated that an accuracy of +_ 0,02 mg/1, expressed
as one standard deviation, can be achieved when analyzing Lake Michigan open
water, inshore, and harbor samples by distillation. TJhen samples from the
above waters are analyzed employing the AutoAnalvzer, an accuracy of +_ 0001
mg/1 can be achieved0 The precision that can be obtained by distillation is
+ 0.01 mg/1 (one standard deviation), and when employing the AutoAnalvzer,
the precision is better than this'value.
2. The above distillation results have been obtained by modifying the
method for ammonia nitrogen, as given in Standard Methods, 12th Edition, to
comply strictly with the procedure as presented in the Appendix of this
report.
3. The AutoAnalvzer procedure was demonstrated tc be superirr (more
accurate with greater precision) to the distillation procedure for the
relatively uncontaminatsd lake waters.
U. Of the 11 participating laboratories, the majority was able to
achieve the above accuracy using the distillation procedure. These results
were not achieved-on a routine basis, but un-'er conditions requiring extra-
ordinary care and precautions. Laboratories exposed to extreme air pollution
problems, and those sriall laboratories where all analyses must be performed
in one room, will most likely not be able to obtain these accuracies at the
concentrations under investigation.
5. The limit of detectability using the modified distillation procedure,
as presented herein for the detection of ar.r.or.ia nitrogen in Lake Michigan
waters, appears to be O.C2 mg/1. With the AutoAnalyzer, the" limit appears
to be 0.01 mg/1. It is possible that the sensitivity of the distillation
procedure can be improved, "cwever, such a ncssibility can be verified only
through further applied research.
6. Onlv th:-se results from laboratories shrwing the continuing capability to
analyse at an accuracy of + 0.02 mg/1, or better, should be acce'ct^d in a
monitoring operation "/hen analyzing for ammonia nitrogen levels below J0C6
mg/1.
7. Historic data reporting ammonia nitrogen levels below 001 mg/1
should be evaluated with extreme caution. Ihere appears to be no practical
way to develop a correction factor to apolv to the latter data which would
provide acceptable correlations in the lower concentration ranges.
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18
BIBLIOGRAPHY
1. Standard Methods for the Examination of Water and Wastewater,
12th Edition, American Public Health Association, Inc.,
New York, N.T., 1965.
2. O'Brien, James E. and Fiore, Janece. "Ammonia Determination
b7 Automatic Analysis." Wastes Engineering, July 1962.
3. Jenkins, David. "A Study of Methods Suitable for the Analysis
and Preservation of Nitrogen Forms in an Estaurine Environ-
ment." SERL Report No. 65-13. U.S. Dept. of Health, Education,
and Welfare, Water Pollution Control Administration, San
Francisco, California, August 1965.
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APPENDIX
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20
ANALYTICAL- PROCEDURAL DETAILS FOR THE ANALYSIS OF AMMONIA NITROGEN
The sodium phenate method, using the AutoAnalyzer, is recommended by- the
authors of this report for the analysis of ammonia nitrogen, as are the
methods found in the 12th Edition of Standard Methods_for the Examination
of Water and Vastewater, in particular the distillation-nesslerization
method (pages 186 through 19li), with the modifications presented herein.
1. AutoAnalvzer Procedure - Ammonia Nitrogen
Sodium Phenate Method:
Reagents:
1. Alkaline phenol solution: Dissolve 83 g phenol in 200 ml of
distilled water at room temperature and add 180 ml of 5>-N-
sodium hydroxide solution to the phenol solution. Make up
the volume to 10CO ml with distilled water and store in an
amber bottle; keep in refrigerator when not in use» If the
sodium phenate turns dark, discard it.
2. Sodium hypochlorite solution: $% available chlorine.
3. Sodium nitroprusside, O.CO£?: Dissolve 1 g in 100 ml of
distilled water. Dilute 5 ml of this solution to 1000 ml
with distilled water. Store in refrigerator at li°C.
U. Standards: Dissolve 3.819 g of ammonium chloride in
ammonia-free water, add 0.8 ml cone. .^SO^ and make up to
1000 ml. This solution contains 1 rag of nitrogen per ml.
Appropriate standards are prepared by diluting this stock
solution.
Procedure:
1. Set up the analytical system in accordance with the attached
flow diagram. Allow 30 minutes for instrument to warm up
and establish a base line using distilled water as the sample,
2, Filter turbid samples through a pledget of cotton or membrane
filter, discarding the first 10 ml of filtrate.
3. Arrange five standards, ranging from C-0.75 mg/1, and samples
on the turntable with a single distilled water wash between
each sample and standard.
U. There should be a set of standards placed in each row.
5. Set the cycle time at two minutes and the sample time at
one minute and 59 seconds.
6. All readings made at a magnification of 2X when the NHo-N
levels are very low, and at IX at higher levels.
-------
-------
21
Distillation Procedure - Ammonia Nitrogen
Of all the methods listed in Standard Methods, 12th Edition, the
distillation method is preferred for the determination of ammonia
nitrogen, in trace or appreciable amounts, especially when inter-
ferences of any nature are suspected. The distillates may be
titrated* or nesslerized. Nesslerization of the distillate is the
method of choice for distillates containing 1 mg or less of ammonia
nitrogen, (in the absence of interfering substances, the direct
nesslerization method may be used but this method is reserved for
samples containing ammonia nitrogen in excess of 0.2 mg/1.)
Nesslerization Method:
Apparatus:
1. All-glass distilling apparatus with 800-2000 ml carc-civr flask.
2. Spectrophotometer or filter photometer for use at 1:25r_i,
and providing a light path of 1 cm or longer.
3. Nessler tubes 50 ml tall form.
Reagents:
»
All reagents prepared in ammonia-free distilled water.
1. Ammonia-free water may be prepared by redistilling do*.iiled
water containing 1 ml of concentrated sulfuric arid ;:er liter,
or as stated in Standard Methods, p. 189.
2. Phosphate buffer solution, p. 189-190.
3. Stock ammonium solution, p. 190.
U» Standard ammonium solution, p. 190.
5. Neutralization reagent l.ON sodium hydroxide, p. 1?Q.
6. Neutralization reagent l.ON sulfuric acid, D. 150.
7. Nessler reagent, p. 190.
8. Rochelle salt solution, p. 19U.
9. Sulfuric acid 0.02N, p. 50.
10. Boric acid solution, p. 190.
Procedure:
1. Preparation of equipment: Add 500 ml of distiller -•---«,
10 ml of phosphate buffer solution, and a few boil'.r.,; .. .: s
to a 800 ml flask. Steam out the distillation appara-.u.^
until the distillate shows no trace of ammonia, p. I~~,
Sample preparation: Use 500 ml of sample. Neutral'.--
pH of about 6.6 using a pH meter for the measurement.
10 ml of the phosphate buffer, mix and check the pH ;:':
should now be 7.1i. If it is not, add another 10 ml ol
buffer solution and again check the pH, p. 190.
* If titration is used, collect in boric acid as given in
Standard Methods, p. 191, Sec. li.lt and p. LiOlj, Sec. h.?.
-------
-------
22
Distillation :
Distill over 300 ml into a £00 mi irlenmeyer flask containing
50 ml of 0.02N sulfuric acid for nesslerization (or £0 ml 2%
boric acid if titration is used). Distill at the rate of 6-10
ml/min. Lower the collected distillate free of contact with
the delivery tube, and continue distillation during the last
minute or two to cleanse the condenser and delivery tube.
Preliminary nesslerization:
Nesslerize a 50 ml portion of the distillate to determine if
the concentration of ammonia in the distillate is in the
satisfactory reading range for the colorinetric equipment used.
For example, for a Spectronic 20 with a 1 inch cell, the satis-
factory range is 0.10 to 10L mg/1. If the concentration is below
0,10 mg/1, the ammonia in the distillate should be concentrated
by boiling. If the ammonia nitrogen is above 1,0 mg/1, the
titrimetric method may be employed for measuring the ammonia in
the distillate, or bv nesslerization of a suitable aliquot.
Concentration of ammonia in the distillate :
Since the distillate is in the presence of acid (C002N sulfuric
acid) it may be boiled with no loss of ammonia.
1. Add boiling chips to the flask and boil on a hot plate
to a volurr.e of about 100 or £0 ml depending upon results
from the preliminary nesslerization.
2. If the concentration of ammonia is less than 0.10 mg/1, the
sample should be concentrated by evaporation. Transfer to
50 ml nessler tubes and bring the volume up to the JO ml
mark with anncnia-free v;ater. The concentration factor in
this case is 222 or 10.
3. If the concentration of ammonia nitrogen in the distillate
is in the satisfactory range (0.10 to lob. rg/l) the
concentration factor is $00 or l»i;3«
~
To each 50 ml sample or aliquot in the nessler tubes, add
loO ml of nessler reagent and mix by inverting the tube
six times. Allow for color developnent at least 10 minutes,
but not more than 20 minutes if the ammonia nitrogen is very
low, but in any event run the samples and the standards
the same length of time. Read at Ii25 nn wavelength. The
occasional formation of cloudiness following nesslerization
may be prevented by the use of two drops of Rochelle salt
solution (Standard Methods, p. 189).
-------
-------
23
Preparation of standard curve:
*
The standard curve should be prepared under the sane conditions
as the samples. The blank (reagent blank) and the appropriate
aliquot of standards - diluted to £00 ml each - are distilled
in the same manner as the samples. The 30O ml distillate and
50 ml of 0.02 N sulfuric acid are brought up to the 500 ml
volume before talcing the 50 ml portion for nesslerization. The
phosphate buffer, Rochelle salt solution, and nessler reagent
are used as for the samples.
Sample Preparation and Collection
Several NHo-N standards were prepared by the Great Lakesrlllinois
River Basins Project and Indiana State Board of Health. Standards
were prepared from anhydrous ammonium chloride dried at 100°C and
dissolved in ammonia-free water. Ammonia-free water was prepared
by redistilling distilled water containing sulfuric acid.
Lake samples were collected from the water intakes of the Gary-
Hobart Water Corporation and Chicago's Central District Filtration
Plant. These sairples were collected in three five-gallon carboys
and then blended in a small drum or tank in o*rder to obtain a
homogeneous sample. One-half gallon samples were drawn off for
the 11 laboratories and the remainder of the sample was spiked
with the standard NHo-N solution. A second set of 11 half-gallon
samples were drawn off for the participating laboratories.
River sarrples were collected from Indiana Harbor Canal at 15>ls*
St. These samples were collected and prepared in the sane manner
as the lake samples. All samples were preserved with-0.8 nl of
cone. HSO per liter.
Ij. Analysis
The datawara analvaed according to the method of least squares to
obtain one and two standard deviations. A line of best fit was
plotted on probability paper. The slope of this line was based on
one standard deviation: the mean (or known) value, plus and mir.us
3h percent, plotted at 16 and 8Ii percent on the abscissa.
The standard deviations were plotted in relation to the known amount
of ammonia nitrogen, or in relation to the mean, in the case of lake
and river samples. The standard deviations are indicated by broken
lines and the known amounts or means are indicated by a solid line.
Probability curves like those shown in Figures 9-lii were prepared
for each individual sample.
-------
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* Data deleted; ( ) number of results deleted before
final computation
## GRW-P Gary Raw Water, Preserved
GRW-NP Gary Raw Water, Not Preserved
SDFP-EW-P South District Filtration Plant, Haw Water, Preserved
SDFP-RW-NP South District Filtration Plant, Raw Water, Not Preserved
Lake Lake Michigan water taken at C-ary-Hobart Water Corp. Raw
Water Intake
River Indiana Rarbor Canal water taken at l|?lst St0
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