Application Note

AN 5391
Rev. 2

Kjeltec™ 8100, 8200 and 8400

Determination of Total Kjeldahl Nitrogen (TKN) in Waste
Water Using Kjeltec 8000 Series According to ISO 5663

TKN (Total Kjeldahl Nitrogen) is the parameter

which usually determined in waste water. It's a
key factor regarding the water application area.
The method has detection limit as 0.5 mg N/l.

Application Note 5391 / Rev. 2 Part No. 6007 4158 1(7)

1 Introduction
Kjeldahl Nitrogen is digested by Sulfuric Acid and converted into Ammonium ions. The ammonia
is then liberated by alkali steam distillation and is fixed by boric acid solution. The Ammonium -
Boric acid receiver solution formed is determined by titration with standardized Hydrochloric Acid.

2 Scope and Field of Application

Sample types:
• Liquid samples: water, waste water
Detection limit: 0.5 mg N/l

3 Principle
Mineralization of the sample to form ammonium sulfate, from which ammonia is liberated and
distilled for subsequent determination by titration.
Conversion of the nitrogen compounds responding to the test to ammonium sulfate by
mineralization of the samples with sulfuric acid, containing a high concentration of potassium
sulfate in order to raise the boiling point of the mixture, in the presence of selenium which acts as a
catalyst.
Liberation of ammonia forms the ammonium sulfate by the addition of alkali and distillation into
boric acid/indicator solution.
Determination of ammonium ion in the distillate by titration with standard acid.

4 Safety Precautions
The use of this application note involves the use of hazardous reagents, operations and equipment.
This application note does not address all the safety risks associated with its use.
Wear gloves (PPE’s) and eye protection while handling sulfuric acid, Hydrochloric Acid, Boric
Acid, and Sodium Hydroxide.
Follow safety instruction of instrument user manuals and Material Safety Data Sheets.

8000190a

Application Note 5391 / Rev. 2 Part No. 6007 4158 2(7)

5 Apparatus
5.1 Foss Kjeltec systems: Kjeltec 8100 Distillation unit,
Kjeltec 8200 Auto distillation unit or Kjeltec 8400
Analyser

5.2 Tecator Line Digestor 2520/2508 or Labtec Line

DT2520/2508 Digestor

5.3 Tecator Scrubber or Labtec Scrubber (Optional)

5.4 250 ml digestion tube

5.5 25 ml burette (or equivalent)

5.6 250 ml Erlenmeyer flask

5.7 Analytical balance

5.8 Pipette-50 ml Capacity

5.9 Boiling rods

6 Reagents
During the analysis, use only reagents of recognized analytical grade.

6.1 98 % Sulfuric acid. The highest purity should always

be used.

6.2 Kjeltabs Se/3.5 (or 7g K2SO4 + 7mg Se)

6.3 40% w/v Sodium Hydroxide

6.4 Titrant
0.005 N or 0.01 N or 0.1 N Hydrochloric Acid (standardized)
Note:
The normality of the titrant is required to 4 decimal places.

6.5 Receiver Solution

• Boric acid 1% with bromocresol green/methyl red indicator solution - for auto titration in
Kjeltec 8400
• Boric acid 4% with bromocresol green/methyl red indicator solution - for manual titration

Application Note 5391 / Rev. 2 Part No. 6007 4158 3(7)

In order to obtain accurate results the receiver solution is adjusted so that a small (0.05 - 0.15 ml)
positive blank is obtained when running a blank sample.

6.5.1 Indicators
Prepare 100 ml of bromocresol green solution (100 mg in 100 ml 95% methanol or ethanol) and 70
ml of methyl red solution (100 mg in 100 ml 95% methanol or ethanol).

6.5.2 Preparation
The 1% boric acid receiver solution is prepared by dissolving 100g boric acid in 5-6 l very hot
deionised water. Mix well until boric acid crystals are completely dissolved. Then add more hot
deionised water to a volume of 9l, mixing well. Cool the solution to room temperature and add the
two indicators as prepared above. Dilute the solution to 10 l with deionised water and mix well.
The 4% boric acid receiver solution is prepared by dissolving 400g boric acid in 5-6 l very hot
deionised water. Mix well until boric acid crystals are completely dissolved. Then add more hot
deionised water to a volume of 9l, mixing well. Cool the solution to room temperature and add the
two indicators as prepared above. Dilute the solution to 10 l with deionised water and mix well.

6.5.3 Adjustment
Adjustment of the boric acid 1 or 4% is made by the following procedure:
1. Transfer 25 ml boric acid solution to a receiver flask and add 100 ml of distilled water. If the
solution in the flask is still red, titrate with 0.1 M sodium hydroxide solution until a neutral
grey colour is obtained. Calculate the amount of sodium hydroxide solution necessary to adjust
the boric acid solution in the 10 l flask with the formula:
ml 0.1 M alkali = ml titrant x 400 or 1.0 M alkali = ml titrant x 40
2. Add the calculated amount of NaOH solution to the boric acid solution. Mix. When using 0.2 N
HCl titrant acid the addition of 30 ml 0.1 M NaOH usually gives a good adjustment , when
using 0.1 N HCl titrant acid the addition of 15 ml 0.1 M NaOH. The lower concentration of
titrant acid used the less addition (adjustment) with NaOH needs to be done.
3. To check proceed as follows using 25 ml of the boric acid solution. Run a blank. If the value of
this blank is high (0.5 ml of 0.2 M HCl) the boric acid is incorrectly adjusted. This might create
irregular blanks. For correction add HCl directly into the boric acid tank, mix it carefully and
repeat until a reading of 0.05-0.15 ml titrant for blank values is obtained. If a positive blank is
not achieved, add further small quantities of NaOH and repeat the check until a satisfactory
value is achieved.
Note: The addition of alkali is to achieve a positive blank value. This should,
however, be kept between 0.05-0.15 ml titrant, to obtain good repeatability
when testing blanks.

7 Analytical Details
7.1 Titrant
The normality of the titrant is required to 4 decimal places. Perform a reagent blank before each
batch of samples.

Application Note 5391 / Rev. 2 Part No. 6007 4158 4(7)

7.2 Instrument Settings
For details see the user manual.

Model Dilution 40% NaOH Delay SAfE Distil Distillation % Steam

end
condition

8100 70 ml 50 ml 12 s 5s 5 min time 100

8200 70 ml 50 ml 12 s 5s 5 min time 100

8400 70 ml 50 ml 12 s 5s 150 mL auto 100

8 Procedure
Use 250 ml digestion tube with one boiling rod.
Perform at least two reagent blank before each batch of samples

8.1 Sample Preparation

Samples should be collected in Polyethylene or glass bottles. They should be analysed as soon as
possible, or else stored between 2-5°C until analysed. Acidification with Sulphuric Acid to pH <2
may also be used as an aid to preservation, provided that possible contamination of the acidified
sample by adsorption of any atmospheric ammonia is avoided.
If the approximate Nitrogen concentration of the sample is known, the test portion volume can be
selected according to:

Nitrogen concentration / mg/l Volume used / ml

up to 20 100

20 - 50 50

50 - 100 25
Table 1 Sample Amount for Different Concentrations
Note:
Interference may occur if the Nitrate concentration in the sample is 10 times
or higher than the TKN level. Use an anion exchange resin (chloride form) to
remove the Nitrate prior to the TKN analysis.

8.2 Digestion
Add 2 Kjeltabs Se/3.5 and 12 ml Sulfuric Acid. Put boiling rod with open end downwards in place,
position the exhaust and turn on the aspirator or scrubber. Digest for 70 minutes in a preheated
block digester at 420°C. Depending on sample content, foaming can occur. If this is the case, the
samples have to be heated slowly.
Using large sample volumes it is advisable to start digestion at ~ 250°C and slowly raise
temperature to 420°C. Remove rack with exhaust and leave to cool for at least 15 minutes.

Application Note 5391 / Rev. 2 Part No. 6007 4158 5(7)

8.3 Distillation and Titration
In Analyzer, part of this is performed automatically.

1. Dilute sample with 70 ml H2 O. In Auto Distillation Unit and Analyzer, part of this is
performed automatically
2. Add 30 ml of receiver solution to the receiver flask.
3. Add 50 ml 40% Sodium Hydroxide solution to the tube.
4. Set steam pressure to 100 %.
5. Allow reaction to settle (delay or safe)
6. Distil for the prescribed time/volume (see above).
7. Remove sample from the distillation unit and titrate distillate with standardized HCl.
Note:
For levels below 10 mg/l the Boric acid should be omitted from the receiver
solution.

9 Expression of Results
9.1 Calculation
(T − B) × N × 14.007 × 1000
Results mg N/l =
Volume (ml) of sample

T = Volume of titrant for sample (ml)

B = Volume of titrant for blank (ml)
N = Normality of titrant

9.2 Method Performance Data

As waste water is not used, ammonium sulfate solution is used instead. Four kinds of solutions with
different concentration are used. The results are shown in Table 2.
Kjeltec 8460, Recovery: >99.5 %

Sample Type Solution 1 Solution 2 Solution 3 Solution 4

Concentration (mg TKN/l) 11.14 34.48 131.58 493.51

11.25 33.87 133.69 482.67
11.45 35.21 138.92 492.31

Mean concentration (mg 11.28 34.52 134.73 487.79

TKN/l)
%RSD 1.39 1.94 2.80 1.22
Table 2 Results of samples containing TKN and relative standard deviations of repeatability
(%RSD)

Application Note 5391 / Rev. 2 Part No. 6007 4158 6(7)

10 References
• ISO 5663 Water quality - Determination of Kjeldahl nitrogen - Method after mineralization
with selenium

11 Revision History
Rev. Date of Issue Revised Material Approved by

1 2016-02-26 New BSun

2 2016-08-19 Debug JICH

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Application Note 5391 / Rev. 2 Part No. 6007 4158 7(7)