Nitrogen, Simplified TKN (s-TKN™) DOC316.53.

01258

s-TKN™ Method Method 102421, 2

®
0 to 16 mg/L TKN TNTplus 880
Scope and application: For water and wastewater. Digestion is required.
1 Revision 1.2, March 2022
2 This method is USEPA equivalent under 40 CFR 136.6 to EPA Method 351.2 and Hach Method 10242, Revision 1.1, January 10, 2013 as
published at 40 CFR part 136.

Test preparation

Instrument-specific information
Table 1 shows all of the instruments that have the program for this test. The table also
shows the adapter and light shield requirements for the applicable instruments that can
use TNTplus vials.
To use the table, select an instrument, then read across to find the applicable information
for this test.
Table 1 Instrument-specific information for TNTplus vials
Instrument Adapters Light shield
DR6000, DR5000 — —
DR3900 — LZV849
DR3800, DR2800 — LZV646
DR1900 9609900 or 9609800 (A) —

Before starting
DR3900, DR3800, DR2800: Install the light shield in Cell Compartment #2 before this test is started.
Review the safety information and the expiration date on the package.
The sample temperature must be 15–25 °C (59–77 °F) for accurate results.
Set the reactor temperature to 120 °C (248 °F). Keep the reactor tube in the reactor for 30 minutes for digestion. Lower
temperatures or shorter digestion time will give incorrect results.
The recommended temperature for reagent storage is 15–25 °C (59–77 °F).
Samples must be preserved with acid as specified in Sample collection and storage on page 2. Make sure to adjust the
pH and temperature before the analysis.
The sample pH must be 3–10 for accurate results.
A small amount of turbidity does not interfere. If there is a large amount of turbidity, let the turbidity settle or filter samples
through a 0.45-µm filter.
Sodium Hydroxide Solution A and Oxidant Tablet B—Make sure to close each bottle immediately after each use.
The 20-mm reaction tube can be used for 7 tests. After each use, clean the tube thoroughly with a brush and water, then
rinse well with high-quality distilled water and let dry.
The nitrite concentration can be determined with nitrite reagents on samples that have not been preserved. Identify the nitrite
concentration (TNT839/TNT840) and subtracted the value from the s-TKN result.
The total nitrogen concentration must be between 1 and 16 mg/L N. The combined nitrate/nitrite concentration must be
between 0.23 and 13.5 mg/L N. Dilute the sample if necessary.
DR1900: Go to All Programs>LCK or TNTplus Methods>Options to select the TNTplus number for the test. Other
instruments automatically select the method from the barcode on the vial.

1
Review the Safety Data Sheets (MSDS/SDS) for the chemicals that are used. Use the recommended personal protective
equipment.
Dispose of reacted solutions according to local, state and federal regulations. Refer to the Safety Data Sheets for disposal
information for unused reagents. Refer to the environmental, health and safety staff for your facility and/or local regulatory
agencies for further disposal information.

Items to collect
Description Quantity
®
s-TKN TNTplus Reagent Set 1
DRB200 reactor with 20-mm wells 1
Pipet, adjustable volume, 1.0–5.0 mL 1
Pipet, adjustable volume, 0.1–1.0 mL 1
Pipet tips 1
Test tube rack 1

Refer to Consumables and replacement items on page 5 for order information.

Sample collection and storage

• Collect samples in clean glass or plastic bottles.
• Adjust the sample pH to less than 2 with concentrated sulfuric acid (approximately
2 mL per liter).
• Keep the preserved samples at or below 6 °C (43 °F) for a maximum of 28 days.
• Let the sample temperature increase to room temperature before analysis.
• Before analysis, adjust the pH to 7 with 5 N sodium hydroxide solution.
• Correct the test result for the dilution caused by the volume additions.

Test procedure

1. Set the DRB200 reactor 2. Add 1.3 mL of sample, 3. Insert the reaction tube in 4. Keep the reaction tube in
power to on. Set the 1.3 mL of Solution A and 1 the preheated DRB200 the reactor for 30 minutes.
temperature to 120 °C Reagent B tablet in quick reactor. Close the lid.
(248 °F). succession to a dry 20‑mm
reaction tube.
Close the reaction tube
immediately. Do not invert.

2 Nitrogen, Simplified TKN TNTplus Method (16 mg/L)

5. When the timer expires, 6. Invert the reaction tube 7. Use a pipet to slowly add 8. Use a pipet to slowly add
carefully remove the 2-3 times. 0.5 mL of the digested 0.2 mL of Solution D to test
reaction tube from the sample from the 20‑mm vial 1 (red label).
reactor. Let the temperature reaction tube into a test vial
of the reaction tube 1 (red label).
decrease to room
temperature.

9. Quickly tighten the cap 10. Use a pipet to slowly 11. Use a pipet to slowly 12. Quickly tighten the cap
on the vial and invert until add 1.0 mL of undigested add 0.2 mL of Solution D to on the vial and invert until
completely mixed. sample to a test vial 2 test vial 2 (green label). completely mixed.
Immediately continue to the (green label).
next step.

Start

13. Start the reaction time 14. When the timer expires, 15. DR1900 only: Select 16. Insert the test vial 1 (red
of 15 minutes. clean the vials. program 880. Refer to label) into the cell holder.
Before starting on page 1. DR1900 only: Push READ1.
Immediately continue to the
next step.

Nitrogen, Simplified TKN TNTplus Method (16 mg/L) 3

17. Insert the test vial 2
(green label) into the cell
holder. DR1900 only: Push
READ2.
Results show in mg/L Total
N, mg/L NO3–N + NO2–N
and mg/L TKN.

Interferences
High levels of oxidizable organic substances (COD) have an effect on the reagent color
and give high results. Use this test procedure for wastewater only when the COD level is
less than 500 mg/L COD. A light pink color may develop during the reaction. The pink
color does not cause interference with the analysis.
Table 2 shows that the ions were individually examined to the given concentrations and
do not cause interference. No cumulative effects or influences of other ions were found.
Table 2 Non-interfering substances
Interfering substance Interference level
Cd2+ 50 mg/L
Ca2+ 50 mg/L
Cl– 500 mg/L
Cr6+ 5 mg/L
Co2+ 10 mg/L
Cu2+ 50 mg/L
Fe2+ 10 mg/L
Fe3+ 50 mg/L
Pb2+ 50 mg/L
Ni2+ 50 mg/L
NO2– 2 mg/L
K+ 500 mg/L
Ag+ 100 mg/L
Na+ 500 mg/L
Sn2+ 50 mg/L
Zn2+ 50 mg/L

Accuracy check
Standard solution method
Use the standard solution method to validate the test procedure, the reagents and the
instrument.

4 Nitrogen, Simplified TKN TNTplus Method (16 mg/L)

 Items to collect:
• Wastewater Effluent Standard Solution, Mixed Parameter (expected result: 7.56 mg/L
Total N, 4 mg/L NO3–N + NO2–N, 3.56 mg/L TKN)

1. Use the test procedure to measure the concentration of the standard solution.
2. Compare the expected result to the actual result. The Wastewater Effluent Standard
Solution contains a component that adds 1.56 mg/L N to the Total N and TKN values.
This is in addition to the 2 mg/L NH3-N and 4 mg/L NO3-N shown on the label.

Summary of Method
Total Kjeldahl Nitrogen (TKN) is the sum of organic nitrogen and ammonia. In the
simplified TKN method, inorganic and organic nitrogen are oxidized to nitrate by digestion
with peroxodisulfate. The nitrate ions react with 2,6-dimethylphenol in a solution of
sulfuric and phosphoric acid to form a nitrophenol. Oxidized forms of nitrogen in the
original sample (nitrite + nitrate due to sample preservation) are determined in the second
test vial and then subtracted, which results in TKN.
Consumables and replacement items
Required reagents

Description Quantity/Test Unit Item no.

®
Simplified TKN (s-TKN) TNTplus reagent set 1 25/pkg TNT880

Required apparatus

Description Quantity/test Unit Item no.

DRB200 Reactor, 115 VAC option, 9 x 13 mm + 2 x 20 mm, 1 block 1 each DRB200-01

DRB200 Reactor, 230 VAC option, 9 x 13 mm + 2 x 20 mm, 1 block 1 each DRB200-05
Pipet, adjustable volume, 1.0–5.0 mL 1 each BBP065
Pipet tips, for 1.0–5.0 mL pipet 1 75/pkg BBP068
Pipet, adjustable volume, 0.1–1.0 mL 1 each BBP078
Pipet tips, for 0.1–1.0 mL pipet 2 100/pkg BBP079
Test tube rack for 20-mm vials, 20 holes 1 each 2497912
Test tube rack for 20-mm vials, 40 holes 1 each 2497902
Light shield, DR3800, DR2800, DR2700 1 each LZV646
Light shield, DR3900 1 each LZV849

Recommended standards

Description Unit Item no.

Wastewater Effluent Standard Solution, Mixed Parameter, for NH3-N, NO3-N, PO43–,
500 mL 2833249
COD, SO42–, TOC

Optional reagents and apparatus

Description Unit Item no.

Filter membrane, 0.45 micron, 25 mm 100/pkg 2514101

Filter holder, 25-mm, for Luer-type syringe each 246800
Sampling bottle with cap, low density polyethylene, 500 mL 12/pkg 2087079
Sodium Hydroxide Standard Solution, 5.0 N 1L 245053

Nitrogen, Simplified TKN TNTplus Method (16 mg/L) 5

Optional reagents and apparatus (continued)
Description Unit Item no.

Sulfuric Acid, concentrated, ACS 500 mL 97949

Syringe, 10 cc, Luer-Lock tip each 2202400
Digestion vials, 20 mm 5/pkg LZP065

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© Hach Company/Hach Lange GmbH, 2009–2010, 2014, 2017–2019, 2021–2022. All rights reserved. 04/2022, Edition 11