Scribd
Upload
39 views

2007 07 10 Methods Method 351 1 PDF

This document provides a method for determining total Kjeldahl nitrogen in surface and saline waters using an autoanalyzer. The sample is digested with sulfuric acid and catalyst to convert organic nitrogen to ammonium sulfate. It is then neutralized and treated with reagents to form an indophenol blue color, which is measured at 630nm. Accuracy was within 25% for samples containing 1.89-5.81 mg/L nitrogen. Precision was within 1.85 mg/L. The applicable range is 0.05-2.0 mg/L nitrogen and approximately 20 samples per hour can be analyzed.

Uploaded by

linhpic99
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd
39 views

2007 07 10 Methods Method 351 1 PDF

This document provides a method for determining total Kjeldahl nitrogen in surface and saline waters using an autoanalyzer. The sample is digested with sulfuric acid and catalyst to convert organic nitrogen to ammonium sulfate. It is then neutralized and treated with reagents to form an indophenol blue color, which is measured at 630nm. Accuracy was within 25% for samples containing 1.89-5.81 mg/L nitrogen. Precision was within 1.85 mg/L. The applicable range is 0.05-2.0 mg/L nitrogen and approximately 20 samples per hour can be analyzed.

Uploaded by

linhpic99
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd
You are on page 1/ 7

METHOD #: 351.

Approved for NPDES, CWA (Ed. Rev. 1974, 1978)

TITLE:

Nitrogen, Kjeldahl, Total (Colorimetric,


Automated Phenate)

ANALYTE:

CAS # N Nitrogen 7727-37-9

INSTRUMENTATION:

Autoanalyzer

STORET No.

00625

1.0

Scope and Application


1.1

2.0

Summary of Method
2.1

3.0

3.2

Total Kjeldahl nitrogen is defined as the sum of free-ammonia and of organic


nitrogen compounds which are converted to (NH4)2SO4 under the conditions of
digestion which are specified below.
Organic Kjeldahl nitrogen is defined as the difference obtained by subtracting
the free- ammonia value from the total Kjeldahl nitrogen value. Also, organic
Kjeldahl nitrogen may be determined directly byremoval of ammonia before
digestion.

Sample Handling and Preservation


4.1

5.0

The sample is automatically digested with a sulfuric acid solution containing


potassium sulfate and mercuric sulfate as a catalyst to convert organic nitrogen
to ammonium sulfate. The solution is then automatically neutralized with
sodium hydroxide solution and treated with alkaline phenol reagent and
sodium hypochlorite reagent. This treatment forms a blue color designated as
indophenol. Sodium nitroprusside, which increases the intensity of the color, is
added to obtain necessary sensitivity for measurement of low level nitrogen.

Definitions
3.1

4.0

This automated method may be used to determine Kjeldahl nitrogen in surface


and saline waters. The applicable range is 0.05 to 2.0 mg N/L. Approximately
20 samples per hour can be analyzed.

Samples may be preserved by addition of 2 mL of conc. H2SO4 per liter and


refrigeration at 4C. Even when preserved in this manner, conversion of
organic nitrogen to ammonia may occur. Therefore, samples should be
analyzed as soon as possible.

Interferences
5.1

Iron and chromium ions tend to catalyze while copper ions tend to inhibit the
indophenol color reaction.

6.0

Apparatus
6.1

7.0

Technicon AutoAnalyzer consisting of:


6.1.1 Sampler II, equipped with continuous mixer.
6.1.2 Two proportioning pumps.
6.1.3 Manifold I.
6.1.4 Manifold II.
6.1.5 Continuous digester.
6.1.6 Planetary pump.
6.1.7 Five-gallon Carboy fume-trap.
6.1.8 80C Heating bath.
6.1.9 Colorimeter equipped with 50 mm tubular flow cell and 630 nm filters.
6.1.10 Recorder equipped with range expander.
6.1.11 Vacuum pump.

Reagents
7.1

7.2

7.3
7.4

7.5
7.6

7.7

7.8

Distilled water: Special precaution must be taken to insure that distilled water
is free of ammonia. Such water is prepared by passage of distilled water
through an ion exchange column comprised of a mixture of both strongly
acidic cation and strongly basic anion exchange resins. Furthermore, since
organic contamination may interfere with this analysis, use of the resin Dowex
XE-75 or equivalent which also tends to remove organic impurities is advised.
The regeneration of the ion exchange column should be carried out according
to the instruction of the manufacturer.
NOTE 1:
All solutions must be made using ammonia-free water.
Sulfuric acid: As it readily absorbs ammonia, special precaution must also be
taken with respect to its use. Do not store bottles reserved for this
determination in areas of potential ammonia contamination.
EDTA (2% solution): Dissolve 20 g disodium ethylenediamine tetraacetate in 1
liter of distilled water. Adjust pH to 10.5-11 with NaOH (7.4).
Sodium hydroxide (30% solution): Dissolve 300 g NaOH in 1 liter of distilled
water.
NOTE 2: The 30% sodium hydroxide should be sufficient to neutralize the
digestate. In rare cases it may be necessary to increase the concentration of
sodium hydroxide in this solution to insure neutralization of the digested
sample in the manifold at the water jacketed mixing coil.
Sodium nitroprusside, (0.05% solution): Dissolve 0.5 g Na2Fe(CN)5NOC2H2O in
1 liter distilled water.
Alkaline phenol reagent: Pour 550 mL liquid phenol (88-90%) slowly with
mixing into 1 liter of 40% (400 g per liter) NaOH. Cool and dilute to 2 liters
with distilled water.
Sodium hypochlorite (1% solution): Dilute commercial "Clorox"-200 mL to 1
liter with distilled water.. Available chlorine level should be approximately 1%.
Due to the instability of this product, storage over an extended period should
be avoided.
Digestant mixture: Place 2 g red HgO in a 2 liter container. Slowly add, with
stirring, 300 mL of acid water (100 mL H2SO4+ 200 mL H2 O) and stir until cool.
Add 100 mL 10% (10 g per 100 ml) K2SO4. Dilute to 2 liters with conc. sulfuric
acid (approximately 500 mL at a time, allowing time for cooling). Allow 4

7.9

7.10
7.11

hours for the precipitate to settle or filter through glass fiber filter.
Stock solution: Dissolve 4.7193 g of pre-dried (1 hour at 105C) ammonium
sulfate in distilled water and dilute to 1.0 liter in a volumetric flask. 1.0 mL =
1.0 mg N.
Standard solution: Dilute 10.0 mL of stock solution (7.9) to 1000 mL. 1.0 mL =
0.01 mg N.
Using the standard solution (7.10), prepare the following standards in 100 mL
volumetric flasks:
Conc., mg N/L
0.00
0.05
0.10
0.20
0.40
0.60
0.80
1.00
1.50
2.00

8.0

mL Standard Solution/100 mL
0.0
0.5
1.0
2.0
4.0
6.0
8.0
10.0
15.0
20.0

Procedure
8.1

8.2

8.3
8.4

Set up manifolds as shown in Figures 1, 2, and 3.


8.1.1 In the operation of manifold No. 1, the control of four key factors is
required to enable manifold No. 2 to receive the mandatory
representative feed. First, the digestant flowing into the pulse chamber
(PC-1) must be bubble free; otherwise, air will accumulate in A-7, thus
altering the ratio of sample to digestant in digester. Second, in
maintaining even flow from the digester helix, the peristaltic pump
must be adjusted to cope with differences in density of the digestate
and the wash water. Third, the sample pick-up rate from the helix must
be precisely adjusted to insure that the entire sample is aspirated into
the mixing chamber. And finally, the contents of the "Mixing Chamber"
must be kept homogeneous by the proper adjustment of the air
bubbling rate.
8.1.2 In the operation of manifold No. 2, it is important in the neutralization
of the digested sample to adjust the concentration of the NaOH so that
the waste from the C-3 debubbler is slightly acid to Hydrion B paper.
8.1.3 The digester temperature is 390C for the first stage and 360C for the
second and third stages.
Allow both colorimeter and recorder to warm up for 30 minutes. Run a
baseline with all reagents, feeding distilled water through the sample line.
Adjust dark current and operative opening on colorimeter to obtain stable
baseline.
Set sampling rate of Sampler II at 20 samples per hour, using a sample to
wash ratio of 1 to 2 (1 minute sample, 2 minute wash).
Arrange various standards in sampler cups in order of increasing
concentration. Complete loading of sampler tray with unknown samples.

8.5
9.0

Calculation
9.1

9.2

10.0

Switch sample line from distilled water to sampler and begin analysis.

Prepare standard curve by plotting peak heights of processed standards


against concentration values. Compute concentration of samples by comparing
sample peak heights with standard curve.
Any sample that has a computed concentration that is less than 10% of the
sample run immediately prior to it must be rerun.

Precision and Accuracy


10.1

Six laboratories analyzed four natural water samples containing exact


increments of organic nitrogen compounds, with the following results:

Increment as
Kjeldahl-Nitrogen
mg N/liter
1.89
2.18
5.09
5.81

Precision as
Standard Deviation
Kjeldahl-N mg N/liter
0.54
0.61
1.25
1.85

Accuracy as
Bias,
%
-24.6
-28.3
-23.8
-21.9

Bias,
mg N/liter
-0.46
-0.62
-1.21
-1.27

Bibliography
1.

2.

3.

Kammerer, P. A., Rodel, M. G., Hughes, R. A., and Lee, G. F., "Low Level Kjeldahl
Nitrogen Determination on the Technicon AutoAnalyzer". Environmental Science and
Technology, 1, 340 (1967).
McDaniel, W. H., Hemphill, R. N., Donaldson, W. T., "Automatic Determination of
Total Kjeldahl Nitrogen in Estuarine Waters". Presented at Technicon Symposium on
Automation in AnalyticalChemistry, New York, October 3,1967.
B. O'Connor, Dobbs, Villiers, and Dean, "Laboratory Distillation of Municipal Waste
Emuents". JWPCF 39, R 25, 1967.

You might also like