CEN/TF

Date: 2005 -08

TC WI

CEN/TF

Secretariat: DS

Determination of ammonium and nitrate in soil, biowaste and sewage sludge

Einführendes Element — Haupt-Element — Ergänzendes Element

Élément introductif — Élément central — Élément complémentaire

ICS:

Descriptors: Horizontal-Inorg Dr Janssen and R.Koopmann

Document type: European Standard

Document subtype:
Document stage: Working Document
Document language: E
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Foreword
This document is a working document.

This document TF WI has been prepared by CEN/BT/Task Force 151 – Horizontal Standards in the Field of
Sludge, Biowaste and Soil, the secretariat of which is held by Danish Standards.

This document has been prepared under a mandate given to CEN by the European Commission and the
European Free Trade Association, and supports essential requirements of EU Directive(s).

For relationship with EU Directive(s), see informative Annex A, B, C or D, which is an integral part of this
document.

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This standard is applicable and validated for several types of matrices. The table below indicates which ones.

Material Validated for Document

(type of sample, e.g. municipal

sludge, compost)

Sludge Domestic sludge Horizontal Report Desk Study 16

Industrial sludge PrEN 14671

Soil Different soil types ISO 14255 ; ISO 14256

Horizontal Desk Study 16

Soil improvers Fertilized peat Not validated yet

Sediment Not validated yet Not validated yet

Waste Bark humus Horizontal Desk Study 16

Composted biowaste

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Contents Page

Foreword.............................................................................................................................................................................2
Introduction........................................................................................................................................................................5
1 Scope.....................................................................................................................................................................6
2 Normative references ........................................................................................................................................6
3 Terms and definitions .......................................................................................................................................7
4 Safety remarks ....................................................................................................................................................7
5 Principle................................................................................................................................................................7
6 Interferences and sources of errors..............................................................................................................7
7 Reagents...............................................................................................................................................................8
8 Apparatus
8.1 Analytical Balance..............................................................................................................................................8
8.2 Extraction Bottles...............................................................................................................................................8
8.3 Shaking apparatus ............................................................................................................................................ 8
8.4 Filter paper ...........................................................................................................................................................8
9 Sampling and sample pre-treatment .............................................................................................................9
9.1 Sampling...............................................................................................................................................................9
9.2 Sample pre-treatment........................................................................................................................................9
10 Procedure .............................................................................................................................................................9
10.1 Extraction .............................................................................................................................................................9
10.2 Filtration................................................................................................................................................................9
10.3 Analysis of natural samples ............................................................................................................................9
10.4 Calibration..........................................................................................................................................................10
10.5 Blank determination ........................................................................................................................................10
10.6 Dublicate determination .................................................................................................................................10
11 Expression of results ......................................................................................................................................10
11.1 Method of calculation......................................................................................................................................10
11.2 Expression of results ......................................................................................................................................10
12 Test report..........................................................................................................................................................11
13 Performance characteristics ................................................................................................................... 11-15
Annex A (informative) Validation of methods ..........................................................................................................16
Annex B (informative) The modular horizontal system.........................................................................................17
Annex C (informative) Information on WP xx and the project Horizontal.........................................................18
Bibliography.....................................................................................................................................................................19

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Introduction
This document is developed in the project 'Horizontal'. It is the result of a desk study "DS 16: Determination of
total phosphorus, total nitrogen and nitrogen fractions" and aims at evaluation of the latest developments in
assessing total nitrogen in sludge, soil, treated biowaste and neighbouring fields. After discussion with all
parties concerned in CEN and selection of a number of test methods described in this study the standard has
been developed further as an modular horizontal method and has been validated within in the project
'Horizontal' .

A horizontal modular approach is being investigated and developed in the project 'Horizontal'. 'Horizontal'
means that the methods can be used for a wide range of materials and products with certain properties.
'Modular' means that a test standard developed in this approach concerns a specific step in a test procedure
and not the whole test procedure (from sampling to analyses).

The use of modular horizontal standards implies the drawing of test schemes as well. Before executing a test
on a certain material or product to determine certain characteristics it is necessary to draw up a protocol in
which the adequate modules are selected and together form the basis for the test procedure.

The other horizontal modules that will be available in due time are to be found in the informative annex [xxx]
which contains a brief overview of the modules that will be worked out in the project 'Horizontal.'

The texts of the chapters 1 to 12 are normative; annexes are normative or informative, as stated in the top
lines of the annexes.

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1 Scope
This standard describes pretreatment and extraction methods for the nitrogen fractions ammonia and nitrate in
soil, sludge, biowaste and related waste. The extraction method is suitable for moist and air dried samples.
The determination of the nitrogen fractions can be done manuelly or by automated methods.

2 Normative references
This European Standard incorporates by dated or undated reference, provisions from other publications.
These normative references are cited at appropriate places in the text and the publications are listed
hereafter. For dated references, subsequent amendments to or revisions of any of these publications apply to
this European Standard only when incorporated in it by amendment or revision. For undated references the
latest edition of the publication referred to applies (including amendments).

ISO 11464 Soil quality – Pretreatment of samples for physico-chemical analysis

ISO 11465 Soil quality – Determination of dry matter and water content on a mass basis – gravimetric method

EN 12880 Characterisation of sludge – Determination of dry residue and water content

CEN/TC 292 WI 29292030 Characterisation of waste – Preparation of test portions from the laboratory sample

prEN 14671 Characterisation of sludges – Pretreatment for the determination of extractable ammonia using

2 mol/l potassíum chloride

ISO 14256-1 Soil quality – Determination of nitrate, nitrite and ammonium in field-moist soils by extraction with
potassium chloride solution – part 1: manual method

ISO 14256-2 Soil quality – Determination of nitrate, nitrite and ammonium in field-moist soils by extraction with
potassium chloride solution – part 2: automated method

E DIN 19746 Soil quality – Determination of mineral nitrogen (nitrate and ammonium) in soil profiles (Nmin
laboratory method)

EN 11732 Water quality – Determination of ammonium nitrogen by flow analysis (CFA and FIA) and
spectrometric detection
ISO 5664 Water quality - Determination of ammonium: distillation and titration method

EN ISO 13395 Water quality – Determination of nitrite nitrogen, nitrate nitrogen and the sum of both by flow
analysis (CFA and FIA) and spectrometric detection

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3 Terms and definitions

For the purpose of this European Standard, the following definition applies:

3.1

Nitrogen fractions

Amount of ammonium-nitrogen and nitrate-nitrogen that is released after single or repeated 1 M KCl
extraction of the sample.

3.2

Dry residue

Dry mass portion of the sample obtained after the specified drying process. It is expressed as percent (EN
12880:2000)

4 Safety remarks
Waste and sludge samples may contain hazardous and inflammable substances. They may contain
pathogens and be liable to biological action. Consequently it is recommended that these samples should be
handled with special care. National regulations should be followed with respect to microbiological hazards with
this method.

5 Principle
An aliquot of the homogenised moist material is shaken for one hour with 1 mol/l potassium chloride solution
at room temperature. The ratio of extractant to material is ten to one. Repeated extractions are recommended
for some materials (9.1) The extraction solution is filtered and the nitrogen fractions are analysed by flow
injection analysis (FIA, EN 11732, ISO 7150-2) or continous flow analysis (CFA, EN 14256-2, EN 11732, ISO
7150-2) or by manual methods as distillation and titration (ammonia, ISO 5664) or spectrophotometric method
(ammonia, nitrate, nitrite, EN 14256-1, ).

6 Interferences and sources of errors

The deep frozen homogenised test sample is directly transferred to the extraction bottel and the potassium
chloride solution if change in the content of the nitrogen fractions is to be suggested. Drying of the material ,
even rapid microwave drying will result in change of content especially of ammonia. Drying is not subject
within this standard. Data for dried samples have been integrated for the process of validation and exchange
of samples.

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7 Reagents
All reagents have to be of recognized analytical grade

7.1 Potassium chloride c(KCl) = 1 mol/l. Dissolve 373 g of potassium chloride, dried at 105 ° C, in
approximately 3 litres of water and dilute to 5 litres with water.

8 Apparatus
Usual laboratory equipment is needed

8.1 Analytical balance with an accuracy of 0,05 g

8.2 Wide necked glass or plastic bottles with secure stopper or caps, nominal volume 250 or 500 ml or
other. The material must not adsorb ammonia, nitrate or nitrite or contaminated with this species.

8.3 Shaking apparatus; End-over-end shaker, frequency 30 min–1 to 40 min–1

8.4 Filter paper, free of nitrogen fractions

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9 Sampling and sample pre-treatment

9.3 Sampling

Sampling should be carried out in accordance with EN yyyy:2003 (Horizontal standard module(s) for sampling
of sludge, soil and waste).

Samples should be stored in suitable containers with an appropriate closure material such as PTFE.

9.4 Sample pre-treatment

All samples shall be pretreated according to the special standard in the fields of soil, sludge, biowaste and
related waste. The samples shall be analysed as soon as possible. The samples can change composition
through biological and/or chemical activity. The samples shall be protected from being warmed up during the
sampling procedure. The transportation to the laboratory shall be organised in such a way that no warming up
occurs. Transportation in a cool box is recommended. If the samples are analysed within three days it is
enough to store them at 4 °C, otherwise they should be stored at -20 °C (deep-frozen), which enables
storing for several weeks, without any significant change in the content of mineral nitrogen. It is advantageous
to homogenise the moist sample, to divide it into the test sample before storing them at -20 °C .

When the content of mineral nitrogen is determined in deep-frozen samples, the temperature and the duration
of the thawing process have to be controlled. The samples can be thawed at room temperature, if they are
homogenised and extracted within 4 h after beginning of thawing. Thawing at 4 °C is also possible, but the
thawing period should not exceed 48 h.

10 Procedure
10.1 Extraction

Transfer a known weight of the homogenised test sample (equal to 1,0 to 5,0 g dry mass) to an extraction
bottel (5.3), add potassium chloride solution (4.2) in the ratio test sample (dry mass) to extraction solution of
one to ten (m/V). Replace the bottel cap and place the extraction bottle to the shaking apparatus (5.4). Shake
the extraction bottel for 1 hour at room temperature. A minimum of one repetition of extraction after filtration is
necessary for dry soil samples < 250 µm and fresh or dry biowaste and sewage sludge samples.The amount
of test sample is related to the homogenising procedure. Take care, that the test sample is a homogeneous
part of the collected sample and the laboratory sample.

10.2 Filtration

Filter the extraction solution through the filter . Collect the filtrate for determination of the nitrogen fractions in a
volumetric flask and fill up to volume with potassium chloride solution. Centrifugation is recommended for
samples , which are subjected to repeated extractions. The analysis of the nitrogen fractions shall be done as
soon as possible. For the high concentration of potassium chloride avoids biological activity, the extracts can
be stored at 4 °C for 30 days.

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10.3 Measuring

Analysis is done according to EN ISO 11732, ISO 14256-2, ISO 14256-1, ISO 5664, EN ISO 14911. State of
the art is to use the flow injection analysis (FIA, reference method) or the continuous flow analysis (CFA,
reference method) , their desciption is presented in the standard EN ISO 11732 and ISO 14256-2

10.4 Calibration

Calibration of the analytical part is done according to the descriptions given in standards in 10.3. using
ammonium and nitrate in inorganic salts, e.g. ammoniumchloride and potassiumnitrate.

10.5 Blank determination

Carry out at least two blank determinations in each series and use the average blank value for subsequent
calculations. Blank determinations are carried out by using 1M KCl without sample addition throughout the
whole procedure.

10.6 Duplicate determination

Analyse two individual test samples of each homogenised sample submitted for analysis. Establish a control
limit for the difference between results for the two sub-samples based for example on precision data in Annex
A or on laboratory precision data.

11 Expression of results

11.3 Method of calculation

The calculation is described in the mentioned standards.

11.2 Expression of results

The results of extractable ammonium, nitrate or nitrite are expressed in g/kg dry mass. The dry mass is
determined according to the related standard.

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12 Test report
The test report shall contain the following information:

a) a reference to this European Standard including its date of publication;

b) precise identification of the sample;

c) expression of results, according to 11.2 ;

d) any deviation from this standard, and any facts which may have influenced the result. Where the test is
not carried out in accordance with this standard, reference may only be made to EN xxxx:2003 in the
report in case all deviations from the procedures prescribed in this standard are indicated in the report
stating the reason for deviation.

13 Performance characteristics
Performance data in terms of repeatability and reproducibility .have been determined during desk study 16
using statistical data from 6 repeated measurements of one sample analysed the same day.

Repeatability is expressed by standard deviation in %N and as relative standard deviation s rel

Sample NH4-N (mg/kg) S rel % NO3-N (mg/kg) S rel %

Soil K1 0,23 17 < LOQ < LOQ

Soil K3 0,11 18 0,11 18

Biowaste KG 154 1,4 31 2

Biowaste KF 85 13 < LOQ < LOQ

Sludge K19 685 2 < LOQ < LOQ

Sludge K20 697 2 < LOQ < LOQ

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Variation of Extraction conditions – Repetition of extraction

Fresh Sample: Biowaste CW KG + addition of KNO3 and NH4Cl

KCl Ratio: sample / Shaking time NH4-N

extractant
( hours) % Recovery

2M 1:5 1x1h 71 +/- 6

2M 1 : 10 1x1h 85 +/- 7

2M 1.5 2x1h 87 +/- 7

2M 1 . 10 2x1h 92 +/- 7

1M 1:5 1 x 1h 66 +/- 6

1M 1 : 10 1 x 1h 80 +/- 7

1M 1 : 10 2 x 1h 81 +/- 7

1M 1 . 10 1 x 2h 76 +/- 6

1M 1 : 10 1 x 1h + 16h ** 60 +/- 6
stand at 20°C

Recovery for nitrate was > 87 % in all samples investigated

Recovery was estimated by addition of 50 mg/l NH4-N and NO3-N to the extraction solution

** The filtrate was left at room temperature over night to test the stability of filtrates

Each data point represents the mean of 2-6 extractions

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Analysis of dry samples

Dry sample: soil SO K3

KCl Ratio: sample / Shaking time NH4-N NO3-N

extractant
( hours) (mg/kg) (mg/kg)

1M 1 : 10 1x1h 0,20 0,32

1M 1 : 10 2x1h 0,27 0,32

1M 1 : 10 3x1h 0,27 0,32

Dry sample: soil WEPAL standard (< 2 mm)

KCl Ratio: sample / Shaking time NH4-N NO3-N

extractant
( hours) (mg/kg) (mg/kg)

1M 1 : 10 1x1h 2,25 2,37

1M 1 : 10 2x1h 2,50 2,49

1M 1 : 10 3x1h 2,57 2,52

Dry sample: soil SO 9, < 125 µm

KCl Ratio: sample / Shaking time NH4-N NO3-N

extractant
( hours) (mg/kg) (mg/kg)

1M 1 : 10 1x1h 5,64 3,82

1M 1 : 10 2x1h 6,58 4,15

1M 1 : 10 3x1h 6,85 4,30

Each data point represents the mean of 2-4 extractions; S rel: < 2%

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Stability of filtrates in 1M KCl

Test portion 2 of CW KF (1 : 10) in 1M KCl

T = 4 °C

Time NH4-N NO3-N

(days) mg/kg mg/kg

0 46,2 +/- 3,7 < LOQ

22 46,3 +/- 3,7

30 45,4 +/- 3,6

42 46,4 +/- 3,7

56 45,6 +/- 3,6

T = - 18°C

Time NH4-N NO3-N

(days) mg/kg mg/kg

0 46,2 +/- 3,7 < LOQ

22 46,4 +/- 3,7

30 46,0 +/- 3,7

42 45,3 +/- 3,6

56 43,4 +/- 3,5

LOQ = 0,5 mg/l test solution (CFA)

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Test mixture of KNO3 and NH4Cl in 1M KCl

T = 4 °C

Time NH4-N NO3-N

(days) mg/kg mg/kg

0 46,6 +/- 0,5 44,8 +/- 0,5

22 46,1 +/- 0,5 44,7 +/- 0,5

30 46,7 +/- 0,5 46,9 +/- 0,5

42 44,4 +/- 0,4 44,4 +/- 0,4

56 41,2 +/- 0,4 47,4 +/- 0,5

T = - 18 °C

Time NH4-N NO3-N

(days) mg/kg mg/kg

0 46,6 +/- 0,5 44,8 +/- 0,5

22 46,2 +/- 0,5 46,1 +/- 0,5

30 46,9 +/- 0,5 45,4 +/- 0,5

42 44,9 +/- 0,4 44,7 +/- 0,5

56 40,5 +/- 0,4 46,5 +/- 0,5

Remark: mg/kg refers to a hypothetical amount of 5g sample in 50 ml, though the test mixture was extracted
without any sample matrix. Extraction, analysis in CFA and storage was performed as described for sample
extraction and analysis.

Standard deviation was < 1 % for the analysis of both ions in artificial solution, as calculated by two
repeated measurements in CFA

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Annex A (informative)

Validation of methods

The extraction procedure is not validated. This has to be done for all materials in future. The determination of
nitrogen species is validated in EN ISO 11732.

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Annex B
(informative)

The modular horizontal system

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Annex C
(informative)

Information on WP xx and the project Horizontal

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Bibliography

Page, A.L. et al. (1982): Methods of soil analysis, Part 2, American Society of Agronomy and Soil Science of
America, Madison, WI.

Lickfett, T., Merkel, D., von Hörstein, D., Przemeck, E., Lücke, W. (1996): Konservierung von Nmin-Proben
durch Mikrowellentrocknung? Agrobiol.Res. 49, 169-178

ISO 11464 Soil quality – Pretreatment of samples for physico-chemical analysis

ISO 11465 Soil quality – Determaination of dry matter and water content on a mass basis – gravimetric
method

EN 12880 Characterisation of sludge – Determination of dry residue and watercontent

CEN/TC 22 WI 29292030 Characterisation of waste – Preparation of test portions from the laboratory sample

prEN 14671 Characterisation of sludges – Pretreatment for the determination of extractable ammonia using 2
mol/l potassíum chloride

ISO 14256-1 Soil quality – Determination of nitrate, nitrite and ammonium in field-moist soils by extraction with
potassium chloride solution – part 1: manual method

ISO 14256-2 Soil quality – Determination of nitrate, nitrite and ammonium in field-moist soils by extraction with
potassium chloride solution – part 2: automated method

E DIN 19746 Soil quality – Determination of mineral nitrogen (nitrate and ammonium) in soil profiles (Nmin
laboratory method)

EN 11732 Water quality – Determination of ammonium nitrogen by flow analysis (CFA and FIA) and
spectrometric detection
ISO 5664 Water quality - Determination of ammonium: distillation and titration method

EN ISO 13395 Water quality – Determination of nitrite nitrogen, nitrate nitrogen and the sum of both by flow
analysis (CFA and FIA) and spectrometric detection

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