BS EN ISO 14238:2013

BSI Standards Publication

Soil quality — Biological

methods — Determination of
nitrogen mineralization and
nitrification in soils and the
influence of chemicals on these
processes (ISO 14238:2012)
BS EN ISO 14238:2013 BRITISH STANDARD

National foreword
This British Standard is the UK implementation of EN ISO
14238:2013. It is identical to ISO 14238:2012. It supersedes BS
7755-4.4.3:1997 which is withdrawn.
The UK participation in its preparation was entrusted to Technical
Committee EH/4, Soil quality.
A list of organizations represented on this committee can be
obtained on request to its secretary.
This publication does not purport to include all the necessary
provisions of a contract. Users are responsible for its correct
application.
© The British Standards Institution 2013. Published by BSI Standards
Limited 2013
ISBN 978 0 580 82333 6
ICS 13.080.30
Compliance with a British Standard cannot confer immunity from
legal obligations.
This British Standard was published under the authority of the
Standards Policy and Strategy Committee on 31 December 2013.
Amendments issued since publication
Date Text affected
EUROPEAN STANDARD EN ISO 14238
NORME EUROPÉENNE
EUROPÄISCHE NORM December 2013

ICS 13.080.30

English Version

Soil quality - Biological methods - Determination of nitrogen

mineralization and nitrification in soils and the influence of
chemicals on these processes (ISO 14238:2012)

Qualité du sol - Méthodes biologiques - Détermination de la Bodenbeschaffenheit - Biologische Verfahren - Bestimmung

minéralisation de l'azote et de la nitrification dans les sols, der Stickstoffmineralisierung und -nitrifizierung in Böden
et de l'influence des produits chimiques sur ces processus und der Einflüsse von Chemikalien auf diese Prozesse (ISO
(ISO 14238:2012) 14238:2012)

This European Standard was approved by CEN on 10 November 2013.

CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this European
Standard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such national
standards may be obtained on application to the CEN-CENELEC Management Centre or to any CEN member.

This European Standard exists in three official versions (English, French, German). A version in any other language made by translation
under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC Management Centre has the same
status as the official versions.

CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,
Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United
Kingdom.

EUROPEAN COMMITTEE FOR STANDARDIZATION

COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG

CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels

© 2013 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 14238:2013 E
worldwide for CEN national Members.
BS EN ISO 14238:2013
EN ISO 14238:2013 (E)

Foreword

The text of ISO 14238:2012 has been prepared by Technical Committee ISO/TC 190 “Soil quality” of the
International Organisation for Standardization (ISO) and has been taken over as EN ISO 14238:2013 by
Technical Committee CEN/TC 345 “Characterization of soils” the secretariat of which is held by NEN.

This European Standard shall be given the status of a national standard, either by publication of an identical
text or by endorsement, at the latest by June 2014, and conflicting national standards shall be withdrawn at
the latest by June 2014.

Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. CEN [and/or CENELEC] shall not be held responsible for identifying any or all such patent rights.

According to the CEN-CENELEC Internal Regulations, the national standards organizations of the following
countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech
Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece,
Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal,
Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom.

Endorsement notice

The text of ISO 14238:2012 has been approved by CEN as EN ISO 14238:2013 without any modification.

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ISO 14238:2012(E)

Contents Page

Foreword............................................................................................................................................................................. iv
Introduction......................................................................................................................................................................... v
1 Scope....................................................................................................................................................................... 1
2 Normative references.......................................................................................................................................... 1
3 Terms and definitions.......................................................................................................................................... 1
4 Principle.................................................................................................................................................................. 2
5 Materials.................................................................................................................................................................. 2
5.1 Soils......................................................................................................................................................................... 2
5.2 Reagents and materials...................................................................................................................................... 3
5.3 Test substance...................................................................................................................................................... 3
6 Apparatus............................................................................................................................................................... 4
7 Procedures............................................................................................................................................................. 4
7.1 Experimental options.......................................................................................................................................... 4
7.2 Treatment of soils................................................................................................................................................. 4
7.3 Incubation of soils................................................................................................................................................ 5
7.4 Sampling of soils for testing............................................................................................................................. 6
7.5 Extraction of soils................................................................................................................................................ 6
7.6 Analyses................................................................................................................................................................. 6
8 Expression of results.......................................................................................................................................... 6
8.1 Basic mineralization test.................................................................................................................................... 6
8.2 Toxicity testing...................................................................................................................................................... 7
9 Test report.............................................................................................................................................................. 8
Annex A (informative) Determination of water-holding capacity of soil............................................................. 10
Bibliography...................................................................................................................................................................... 12

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Foreword

ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies
(ISO member bodies). The work of preparing International Standards is normally carried out through ISO
technical committees. Each member body interested in a subject for which a technical committee has been
established has the right to be represented on that committee. International organizations, governmental and
non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the International
Electrotechnical Commission (IEC) on all matters of electrotechnical standardization.

International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2.

The main task of technical committees is to prepare International Standards. Draft International Standards
adopted by the technical committees are circulated to the member bodies for voting. Publication as an
International Standard requires approval by at least 75 % of the member bodies casting a vote.

Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. ISO shall not be held responsible for identifying any or all such patent rights.

ISO 14238 was prepared by Technical Committee ISO/TC 190, Soil quality, Subcommittee SC 4, Biological methods.

This second edition cancels and replaces the first edition (ISO 14238:1997), which has been technically revised.

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Introduction

Soil consists of both living and non-living components which exist in a complex and heterogeneous environment.
Microorganisms in the soil are mainly responsible for cycling of some nutrients and thus play an essential role
in the maintenance of soil fertility. One of the most important microbial processes in soil is the mineralization
of nitrogen contained in organic forms to ammonium (ammonification) and thereafter to nitrite and nitrate
(nitrification). Clearly, any long-term interference with this process could influence soil fertility.

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BS EN ISO 14238:2013
 BS EN ISO 14238:2013
INTERNATIONAL STANDARD ISO 14238:2012(E)

Soil quality — Biological methods — Determination of nitrogen

mineralization and nitrification in soils and the influence of
chemicals on these processes

1 Scope
This International Standard specifies laboratory procedures for measuring the mineralization and nitrification
of nitrogen by the soil microbiota.

For investigations on the evaluation of soil quality or on effects of contamination, a procedure is given to
measure the rates and extent of N-mineralization in soil or soils of known or unknown quality.

For investigation of the potential toxicity of chemicals to N-mineralization in soils, a simple procedure is given
which allows the impact of single chemicals to be assessed and provides a basis for comparison of the toxicities
of different chemicals.

2 Normative references
The following referenced documents are indispensable for the application of this document. For dated
references, only the edition cited applies. For undated references, the latest edition of the referenced document
(including any amendments) applies.

ISO 10381-6, Soil quality — Sampling — Part 6: Guidance on the collection, handling and storage of soil under
aerobic conditions for the assessment of microbiological processes, biomass and diversity in the laboratory

ISO 10390, Soil quality — Determination of pH

ISO 10694, Soil quality — Determination of organic and total carbon after dry combustion (elementary analysis)

ISO 11260, Soil quality — Determination of effective cation exchange capacity and base saturation level using
barium chloride solution

ISO 11261, Soil quality — Determination of total nitrogen — Modified Kjedahl method

ISO 11274, Soil quality — Determination of the water-retention characteristic — Laboratory methods

ISO 11277, Soil quality — Determination of particle size distribution in mineral soil material — Method by
sieving and sedimentation

ISO 11465, Soil quality — Determination of dry matter and water content on mass basis — Gravimetric method

3 Terms and definitions

For the purposes of this document, the following terms and definitions apply.

3.1
nitrogen mineralization
N-mineralization
microbial degradation of an organic substance containing nitrogen, via the processes of ammonification and
nitrification, to the respective inorganic endproducts, specifically ammonium and nitrate

3.2
ammonification
microbial degradation of organic nitrogen to ammonium

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3.3
nitrification
microbial oxidation of ammonium to nitrite and thereafter to nitrate

3.4
inhibitory dose
IDx
amount of a chemical added to soil that effectively inhibits N-mineralization by a stated percentage, after a
given time, in comparison to an untreated control

EXAMPLE ID25 and ID50 indicate 25 % and 50 % inhibition of N-mineralization, respectively.

4 Principle
The rates or extent of N-mineralization in aerobic soils are determined by measuring the concentrations of
ammonium, nitrite, and nitrate released during mineralization of nitrogen contained in the soil organic matter or
during mineralization of an added nitrogenous organic compound.

The influence of chemicals on N-mineralization is determined by amending soil with a readily degradable source
of organic nitrogen, and measuring the percentage inhibition of product formation in test portions treated with
different quantities of a chemical as compared to an untreated control.

5 Materials

5.1 Soils

5.1.1 Selection of soils

5.1.1.1 Basic mineralization test

For basic tests used for comparing the N-mineralization capacities of different soils or for comparing
N-mineralization in one soil collected at different times of the year, ensure that the choice of soil(s) is consistent
with the purpose of the determination.

5.1.1.2 Toxicity testing

To determine the influence of chemicals on N-mineralization, use a soil with low content (as a mass fraction) of
organic carbon (0,5 % to 1,5 %) and low content of clay.

NOTE Such soil represents a worst-case situation, since adsorption is minimum and availability of the chemical to the
microbiota is maximum. For routine testing, soils with a pH(KCl) less than 5 are not satisfactory, as the rate of nitrification is
likely to be too low to permit a valid assessment of the effects of the chemical on the process. Sandy loam soils and loamy
sand soils are preferred.

5.1.2 Collection, handling and storage of soils

For all tests, the recommendations in ISO 10381-6 for collection, handling and storage of soil shall be followed.

The following information shall be documented:

— date of collection;

— date(s) used in experiments;

— storage conditions, including temperature, moisture content;

— length of storage.

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5.1.3 Characterization of soils

To facilitate interpretation of data and for comparative purposes, the following characteristics shall be determined:

a) physical properties:

— particle size distribution measured in accordance with ISO 11277,

— water content in accordance with ISO 11465,

— water retention characteristic in accordance with ISO 11274 and/or water-holding capacity in accordance
with Annex A;

b) chemical properties:

— pH of soil in accordance with ISO 10390,

— cation exchange capacity in accordance with ISO 11260,

— organic matter content in accordance with ISO 10694,

— total nitrogen content in accordance with ISO 11261.

5.2 Reagents and materials

5.2.1 Quartz sand, fine and clean, of particle size 0,05 mm to 0,2 mm.

5.2.2 Potassium chloride, solution c(KCI) = 1 mol/l

5.2.3 Nitrogeneous substrate, at a concentration of nitrogen in soil of ∼100 mg/kg.

For example:

— lucerne meal, with a C-to-N mass ratio of approximately 16 to 1;

— horn meal, with a C-to-N mass ratio of approximately 16 to 1;

— any other appropriate finely ground organic nitrogen source.

Mineralization of nitrogen may also be measured from the organic matter of the soil. In this case, soil is not
amended with an organic nitrogen source.

For tests in which nitrification alone is of interest, ammonium [as (NH4)2SO4] is an appropriate nitrogen source.

5.3 Test substance

A test substance is only needed when the purpose of the investigation is to determine whether the substance
can influence N-mineralization. Test substances shall be the purest that are commercially available. In many
circumstances, it may be appropriate to test technical-grade or commercial-grade chemicals or mixtures.

NOTE If carriers or formulation ingredients are mixed with the test substance, their influence on N-mineralization (if
any) should be taken into account.

In standard experiments with known test substances, the following data (if applicable) shall be given:

— name (IUPAC);

— structure;

— Chemical Abstracts Service number;

— relative molecular mass;

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ISO 14238:2012(E)

— purity;

— stability in water;

— solubility in organic solvents;

— vapour pressure;

— octanol/water partition coefficient (POW);

— common logarithm of the acid dissociation constant (pKa);

— absorption coefficient (Koc).

6 Apparatus
Usual laboratory apparatus and in particular the following.

6.1 Mechanical shaker.

6.2 Centrifuge or filter paper (nitrate- and ammonium-free).

6.3 Instruments for measurement of concentrations of ammonium, nitrate, and nitrite in soil extracts.

7 Procedures

7.1 Experimental options

7.1.1 Basic mineralization test

To compare N-mineralization capacities in different soils or N-mineralization in one soil collected at different
times of the year, ensure that the design of the experiment and the analyses performed are consistent with the
goals of the experiment.

7.1.2 Toxicity testing

To determine the influence of chemicals on N-mineralization, treat a single microbiologically active soil with at least
five concentrations of the test substance. For convenience, limit the analyses to measurement of the quantities of
nitrate formed in treated and control samples after 0 d and 28 d incubation. Using this simple test design, dose–
response relationships can be established. In some cases, e.g. where soil concentrations are known or can be
predicted by rough screening (as for pesticides), it is possible that dose–response information is not needed,
and an untreated sample and one appropriate concentration of test chemical is sufficient. If the influence of the
chemical is not known, it is recommended that a range-finding test be performed before the final test.

7.2 Treatment of soils

7.2.1 Basic mineralization test

Choose a substrate from the list given in 5.2.3, although the final choice of specific organic substrate used
depends on the purpose of the test. Mix the organic material chosen thoroughly and homogeneously into the
soil. If mineralization of nitrogen from the soil organic matter is being investigated, a nitrogeneous substrate
need not be added.

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7.2.2 Toxicity testing

To determine the influence of chemicals on N-mineralization, use any of the nitrogenous substrates given in 5.2.3.

NOTE Compounds with low C-to-N ratios (i.e. not much higher than 16 to 1) are probably the best choice as little of
the nitrogen released during mineralization is immobilized by the soil microbiota.

Mix the chosen nitrogenous substrate (5.2.3) thoroughly and homogeneously into the soil. Then, divide the soil
into six test portions of equal mass. Mix five of these test portions with different concentrations of the substance
to be tested. Sufficient soil should be prepared so that the soil can later be split into at least three replicates
for each dose. Mix the remaining test portion, but do not add any test chemical (if a carrier is used, mix only
into the soil). The chemical-free test portion serves as the untreated control. If possible, select a concentration
series which allows ID25 or ID50 values to be estimated.

Apply the test substance using an appropriate carrier, for example:

a) in water, depending upon water solubility of the compound;

b) on a solid, e.g. mixed with quartz sand (5.2.1) or with a portion of the soil under investigation.

With many organic chemicals, the soil or sand used as a carrier can be coated with the test chemical by dissolving
it in a solvent. In such cases, the solvent should be removed by evaporation before mixing with the soil.

When water is used to apply the test substance, care should be taken that the water content (as a mass
fraction) does not exceed 60 % of the water-holding capacity or a water pressure of approximately 0,02 MPa.

7.3 Incubation of soils

For N-mineralization investigations, incubate the soils in either of these two ways:

a) as bulk samples of each variant (e.g. soils of different quality or different levels of contamination) or treatment;

b) as a series of individual test portions of each variant or treatment.

When variants are incubated as bulk samples, prepare large quantities of soil and take test portions (e.g. 10 g
to 100 g) during the experiment, as needed. Here, the amounts of soil prepared are determined by the sizes of
the samples taken, the number of replicates used (at least three), and the duration of the experiment. Mix soils
incubated in bulk thoroughly before taking test portions. With large samples, spread the soil out to a depth of
not more than 3 cm to facilitate oxygen transfer. Also mix the soil sample on a weekly basis.

When variants are held as a series of individual test portions, divide each variant into a series of equal test
portions and sacrifice these test portions as needed. In studies with more than one sampling interval, prepare
sufficient test portions to account for all replicates and sampling times.

NOTE 1 The choice of temperatures, water content of the soil and light conditions during incubation depends on the
purpose of the experiment.

For tests to determine the influence of chemicals on N-mineralization, maintain soils at (20 ± 2) °C and a pore
water pressure of approximately 0,02 MPa to the nearest 5 % [(40 ± 5) % to (60 ± 5) % of the maximum water-
holding capacity] in the dark.

NOTE 2 A temperature of (20 ± 2) °C has been chosen as a standard for comparative purposes and because it gives
relatively rapid results. Temperatures outside this range can be used if they are more appropriate (e.g. because of local
conditions or lack of cooling equipment).

In all experiments, vessels holding soils shall allow free exchange of gases. This helps prevent the development
of anaerobic sites which could cause nitrogen loss through denitrification. Minimize water losses from the soil
by incubating soils in covered vessels. Determine the moisture content of the soil at regular intervals, and
replace losses with deionized water.

NOTE 3 Deionized water can be applied to the surface of the samples as a fine spray.

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When the mineralization potentials of different soils are to be compared, the water pressure of the soils shall
be kept as similar as possible, e.g. by adjusting it to the same level of water-holding capacity (40 % to 60 %).

7.4 Sampling of soils for testing

7.4.1 Basic mineralization test

The number of samples and frequency of sampling depend on the purpose of the experiments, but shall
be sufficient to allow for accurate measurement of inorganic nitrogen concentrations. For incubation with an
organic substrate, an incubation time of 28 d is recommended. For incubation without substrate, an incubation
time of 48 d may be necessary. It is possible to carry out intermediate measurements, e.g. after 7 d and 14 d.

7.4.2 Toxicity test

To determine the influence of chemicals on N-mineralization, sample the soils directly after treatment (see 7.2)
(0 d) and after 28 d incubation (see 7.3). In most microbiologically intact, organic nitrogen-amended, untreated
soils, the maximum rate of mineralization is reached within 28 d. Thus, sampling at 28 d usually allows valid
estimations of ID25 or ID50 values. It is possible to carry out intermediate measurements, e.g. after 7 d and 14 d.

7.5 Extraction of soils

Extract ammonium, nitrite, and nitrate from soil samples directly after treatment (see 7.4) by shaking samples
with potassium chloride solution (5.2.2) (5 ml potassium chloride per 1 g dry mass equivalent of soil) at 150 r/min
for 60 min. To optimize extraction, do not fill containers holding soil and potassium chloride solution more than
half full. Remove fine particles of soil from the extracts by filtration through a filter paper or centrifugation using
a centrifuge (6.2). If the measurements are not performed extemporaneously, store particle-free extracts at
(-20 ± 5) °C for up to six months. Where novel analytical methods are used, ensure that interference from
extractants is allowed for.

Test substances that contain high quantities of mineralizable nitrogen can contribute to the quantities of
ammonium, nitrite or nitrate formed. Where high concentrations of such substances are required for toxicity
testing, this should be taken into consideration.

7.6 Analyses
Carry out quantitative analyses of ammonium-N, nitrate-N and nitrite-N.

8 Expression of results

8.1 Basic mineralization test

For basic tests, consider preparing mineralization rate curves using values for the individual nitrogen ions. For
this purpose, to allow nitrogen balances to be established, convert the quantities expressed in milligrams per
kilogram of ammonium ( NH +4 ), nitrite ( NO 2− ), and nitrate ( NO 3− ), determined analytically (7.6), to ammonium-N
( NH +4 −N), nitrite-N ( NO 2− −N), and nitrate-N ( NO 3− −N).

In other cases, especially for comparing mineralization rates in different soils, express N-mineralization values
at each sampling interval as a single value called “Nmin”.

To determine Nmin, use the following equation:

(
 ) ( ) ( ) (
  ) ( ) (
Nmin =  NH +4 − N T + NO 2− − N T + NO 3− − N T  −  NH +4 − N S + NO 2− − N S + NO 3− − N S 
 )
where

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( NH +
4 − NT ) is the concentration of ammonium-N, in milligrams of nitrogen per kilogram of dry soil, at
the time of sampling;

( NO −
2 − NT ) is the concentration of nitrite-N, in milligrams of nitrogen per kilogram of dry soil, at the
time of sampling;

( NO −
3 − NT ) is the concentration of nitrate-N, in milligrams of nitrogen per kilogram of dry soil, at the
time of sampling;

( NH +
4 − NS ) is the concentration of ammonium-N, in milligrams of nitrogen per kilogram of dry soil, at
the start of incubation;

( NO −
2 − NS ) is the concentration of nitrite-N, in milligrams of nitrogen per kilogram of dry soil, at the
start of incubation;

( NO −
3 − NS ) is the concentration of nitrate-N, in milligrams of nitrogen per kilogram of dry soil, at the
start of incubation.

8.2 Toxicity testing

To determine the effects of different concentrations of test substance on N-mineralization to nitrate, compare
the rate of formation of nitrate (i.e. milligrams of NO 3− per kilogram of dry mass of soil per day) found in the
treated samples after 28 d incubation to that found in the untreated control. For the calculation of the rate of
nitrate formation, subtract the mean concentration of nitrate (i.e. mg NO 3− per kg of dry mass of soil) at the start
of the test (day 0) from the mean concentration at the end of the test, and divide the value obtained by 28. If
intermediate measurements have been carried out (e.g. after 7 d and 14 d), the rates during the periods between
these measurements (e.g. 0 d to 7 d, 7 d to 14 d and 14 d to 28 d) should also be calculated. Since nitrogen
balances are not necessary for evaluation of toxicity, do not convert milligrams of NO 3− per kilogram of dry
mass of soil values to ( NO 3− −N) per kilogram of dry mass of soil values.

Calculate inhibition values as a percentage of the control, IDx, for each treatment level as follows:

w
NO 3− ,1
ID x = 100 − × 100
w
NO 3− ,2

where

w is the rate of formation in milligrams of NO 3− per kilogram per day in treated soil;
NO 3− ,1

w is the rate of formation in milligrams of NO 3− per kilogram per day in untreated soil.
NO 3− ,2
After making this simple calculation for each concentration of chemical tested, either

a) prepare a dose–response curve similar to that shown in Figure 1, in which the ID25 and ID50 values for a
hypothetical test substance are 1 mg of chemical per kilogram of dry mass of soil and 10 mg of chemical
per kilogram of dry mass of soil, respectively;

b) use regression analysis of the dependence between the test substance dosed and the content of NO 3− −N.

NOTE On completion of tests carried out with certain chemicals, the treated soil can contain more nitrate than
untreated control samples. This is usually due to mineralization of the nitrogen in microbial cells killed by the chemical.
Other causes can be mineralization of nitrogen from the chemical being tested or stimulation of the mineralization of soil
organic carbon.

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

a) a reference to this International Standard (ISO 14238:2012);

b) soil characteristics (see 5.1.2 and 5.1.3);

c) description of the test substance (if applicable) (see 5.3);

d) soil collection, treatment, incubation, including date collected, length of storage, date(s) used in test(s), method
of treatment (if applicable), amounts of chemical applied (if applicable), incubation conditions (see 7.1 to 7.6);

e) sampling dates, extraction procedures and dates;

f) analytical equipment and methods, detection limits, recovery efficiencies;

g) figures and/or tables of results;

h) evaluation of the results and conclusions drawn (if applicable).

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Key
Y w inhibition of nitrate formation
−
NO 3

X wt concentration of test substance

aID dose resulting in 25 % inhibition of N-mineralization
25
bID dose resulting in 50 % inhibition of N-mineralization
50

Figure 1 — Example of a dose–response curve showing concentrations

of test substance inhibiting N-mineralization to NO 3−

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Annex A
(informative)

Determination of water-holding capacity of soil

A.1 General
The method described in this annex has been found to be appropriate for laboratory samples of soils.

A.2 Apparatus and materials

Usual laboratory equipment and in particular the following.

A.2.1 Glass tube, approximately 20 mm to 50 mm in diameter and at least 100 mm in length.

A.2.2 Water bath, at room temperature.

A.2.3 Filter paper.

A.2.4 Drying oven, set to (105 ± 5) °C.

A.2.5 Balance, capable of weighing with an accuracy of ±0,1 g.

A.3 Method
Plug the bottom of the tube (A.2.1) with filter paper (A.2.3), weigh (A.2.5) the assembly, and after filling with the
soil to a depth of 5 cm to 7 cm, place the tube on a rack in a water bath (A.2.2). Gradually submerge the tube
until the water level is above the top of the soil, but below the upper lip of the tube. Leave the soil sample in the
water bath for about 3 h.

As not all water absorbed by the soil capillary can be retained, the tube containing the sample should be placed
for a period of 2 h on very wet finely ground quartz sand for draining.

Weigh the sample, dry (A.2.4) it to constant mass at 105 °C and reweigh it.

A.4 Calculation of water-holding capacity

Calculate the water-holding capacity, wH 2O,c , as a percentage, using Equation (A.1):

m s − m T − mD
wH 2O,c = ×100 (A.1)
mD

where

wH 2O,c is the water-holding capacity, expressed as a percentage of dry mass, %;

ms is the mass of water-saturated soil plus the mass of the tube plus the mass of the filter paper, in
grams;

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mT is the tare (mass of tube plus mass of filter paper), in grams;

mD is the dry mass of substrate, in grams.

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Bibliography

[1] ISO 11266, Soil quality — Guidance on laboratory testing for biodegradation of organic chemicals in
soil under aerobic conditions

[2] Andersch l., Anderson J.P.E. Influence of pesticides on nitrogen transformation in soil. Toxicol.
Environ. Chem. 1991, 30, pp. 153–158

[3] Bremmer J.M. Nitrogen availability indexes. In: Black, C.A. Methods of soil analysis — Part 2. Madison,
Wl: American Society of Agronomy, 1965, pp. 1324–1345

[4] Henrickson A., Selmer-Olsen A.R. Automatic methods for determining nitrate and nitrite in water and
soil extracts. Analyst (Lond.) 1970, 95 pp. 514–518

[5] Selmer-Olsen A.R. Determination of ammonium in soil extracts by automated indophenol method.
Analyst (Lond.) 1971, 96 pp. 565–568

[6] Stanford G., Smith S.J. Nitrogen mineralization potential of soils. Soil Sci. Soc. Am. Proc. 1972,
36, pp. 465–472

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NO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAW

British Standards Institution (BSI)

BSI is the national body responsible for preparing British Standards and other
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BSI is incorporated by Royal Charter. British Standards and other standardization
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