APPLICATION NOTE
E-K-006-2023/A1
TKN Determination in Water and
Wastewater
Reference: Internal method based on AOAC 973.48 Nitrogen (Total) in Water; EPA 351.3; Standard Method EN
25663, ISO 5663-1984
Tested with VELP Scientifica DKL 20 Automatic Kjeldahl Digestion Unit (Code S30100210), KS 1000 Scrubber
(code F307A0660) and UDK 169 Automatic Kjeldahl Nitrogen Protein Analyzer (code F30200165)
NITROGEN DETERMINATION IN WATER
KJELDAHL METHOD
Introduction
Nitrogen-containing compounds act as nutrients in streams, rivers, and reservoirs. The major routes of entry of nitrogen
into bodies of water are municipal and industrial wastewater, septic tanks, feed lot discharges, animal wastes (including
birds and fish), runoff from fertilized agricultural fields and lawns and discharges from car exhausts. Bacteria in water
quickly convert nitrites [NO 2- ] to nitrates [NO 3- ] and this process uses up oxygen. High nitrates in drinking water can cause
digestive disturbances in people.
From an analytical point of view, the properties of water, wastewater and sludge samples can vary considerably depending
upon their source; in unpolluted waters, low nitrogen levels make it necessary to use large sample volumes, whereas high
concentrations of surface-active agents in some wastewaters can also cause foaming during the initial step of the digestion
so the volume required by the analysis is lower.
Total Kjeldahl nitrogen determination in water, wastewater and sludge according to the Kjeldahl Method
The modern Kjeldahl method consists of a procedure of catalytically supported mineralization of organic material in a
boiling mixture of sulfuric acid and sulfate salt at high temperatures until SO 3 fumes are given off and the solution turns
limpid. During the process, the organically bonded nitrogen is converted into ammonium sulfate.
Alkalizing the digested solution liberates ammonia which is quantitatively steam distilled and determined by titration . This
method covers the determination of total Kjeldahl nitrogen in drinking, surface and saline waters, domestic and industrial
wastes. The procedure converts nitrogen components of biological origin such as amino acids, proteins and peptides to
ammonia, but may not convert the nitrogenous compounds of some industrial wastes such as amines, nitro compounds,
hydrazones, oximes, semicarbazones and some refractory tertiary amines.
For all the nitrogen concentrations in water samples, VELP suggests setting the following method: 60 minutes at 420°C.
The below table shows the indicative sample volume depending on the nitrogen concentration.
Nitrogen concentration (mgN / l) Sample volume (ml)
< 10 75
10-50 50
> 50 25
Sample
Urea diluted solutions with different concentrations of nitrogen: 5, 50 and 100ppm
Sample Digestion
• Stir the sample into a beaker using a VELP magnetic stirrer at 700 rpm;
• Put the proper amount of sample into a 250 ml test tube, by using a pipette.
• For each sample, add in the test tube:
o 2 catalyst tablets KJjtabs VCM (code A00000274; 3.5 g K2SO4, 0.1 g CuSO4·5H20 Missouri)
o Mineralization rod (Code A00000431 pack. 10 pcs) is placed in each test tube with the cava side down,
in order to avoid the formation of bubbles, foam, violent shocks and acid splashes.
o 8 ml concentrated sulphuric acid (96-98%)
• Prepare some blanks with all chemicals except the sample and mineralization rod;
• Connect the Digestion Unit to KS 1000 Scrubber (code F307A0660) to neutralize the acid fumes created during
the digestion phase. For samples with low nitrogen content (< 50 ppm), it is recommended the use of the accessory
code A00000477 Condensation kit for water samples. The accessory is strictly required when the tot al sample
volume in the digestion unit is more than 500 ml.
Example:
Digestion Unit Sample volume for each test tube
DK 6 > 80 ml
DK - DKL 8 > 60 ml
DKL 12 > 40 ml
Copyright © 2023 VELP Scientifica. All rights reserved.
No part of this publication may be reproduced or transmitted in any form or for any purpose without the express permission of VELP .
VELP Scientifica, Italy Tel: +39 039 628 811 Fax: +39 039 628 8120 www.velp.com
NITROGEN DETERMINATION IN WATER
KJELDAHL METHOD
Distillation and Titration
Let the test tubes cool down to 50-60 °C.
Condition the UDK 169 unit by performing the Automatic Check-up and Wash-down in the Menu-System.
Distil the samples by selecting the method “27 – Total nitrogen in water” in the operating manual. Using 0.1N Titrant
solution.
Typical Results on Urea Solutions
The results are automatically calculated by UDK 169 as percentage of nitrogen.
Sample volume (ml) mg N ppm
75 0.39 5.17
75 0.40 5.29
75 0.38 5.12
Average ± SD% 0.39 ± 0.01 5.20 ± 0.09
Nitrogen Expected value: 5ppm
Sample volume (ml) mg N ppm
50 2.49 48.72
50 2.49 49.66
50 2.50 50.02
Average ± SD% 2.49 ± 0.01 49.867 ± 0.15
Nitrogen Expected value: 50 ppm
Sample volume (ml) mg N ppm
25 2.50 99.95
25 2.51 100.37
25 2.51 100.26
Average ± SD% 2.51 ± 0.01 100.19 ± 0.22
Nitrogen Expected value: 100 ppm
Copyright © 2023 VELP Scientifica. All rights reserved.
No part of this publication may be reproduced or transmitted in any form or for any purpose without the express permission of VELP .
VELP Scientifica, Italy Tel: +39 039 628 811 Fax: +39 039 628 8120 www.velp.com
NITROGEN DETERMINATION IN WATER
KJELDAHL METHOD
Conclusions
The determination of TKN in water, and wastewater using VELP DKL 20 and UDK 169 gives reliable and reproducible
results in accordance with the expected range.
Benefits of the Kjeldahl method by using DKL 20 and UDK 169:
• High level of precision and reproducibility
• Maximum productivity
• Worldwide official method
• Reliable and easy method
• Time saving
• Moderate running costs
Connecting the UDK 169 to the VELP Ermes Cloud platform it IS possible to easily monitor and control the distillation
step of the analysis in real-time via PC, smartphone or tablet, improving your laboratory experience.
Moreover, the UDK 169 can be used in combination with the AutoKjel autosampler, for a highly productive system, since
it is capable of autonomously processing up to 24 samples with standard test tubes (250 ml) or 21 Jumbo test tubes (400
ml).
Copyright © 2023 VELP Scientifica. All rights reserved.
No part of this publication may be reproduced or transmitted in any form or for any purpose without the express permission of VELP .
VELP Scientifica, Italy Tel: +39 039 628 811 Fax: +39 039 628 8120 www.velp.com
