NORDIC-BALTIC METHOD

COMMITTEE ON No. XXX

FOOD ANALYSIS Ed. X, 20XX

Title of the method (Calibri Bold 16)

After the title, the first page should include a short description of the method,
e.g, the same text as published about the method on the web page (Calibri 14).
The title should be precise and concise, starting with the analytes/agents, the principle, and
matrices. Examples:

Bacterial count. Determination by direct epifluorescent filter technique (DEFT) in raw minced meat.

Tin (Sn). Determination in foods by inductively coupled plasma mass spectrometry (ICPMS).

NMKL
c/o Institute of Marine Research,
P.O. Box 1870, Nordnes,
NO-5817 Bergen
Norway

NMKL methods and procedures are available for purchase at www.nmkl.org.

NMKL invites all readers and users of the methods and procedures to submit comments and
considerations on its contents to the NMKL secretariat at post@nmkl.org.

©NMKL. The NMKL publications (methods and procedures) are copyright-protected by NMKL.
Neither the publications nor any extract from them may be reproduced, stored in a retrieval system
or transmitted to others in any form or by any means, electronic, photocopying, recording or
otherwise, without prior written permission being secured. Reproduction may be subjected to royalty
payments. Violators may be prosecuted.

This NMKL method has been validated in an interlaboratory study.

If the study has been conducted by another organisation than NMKL that
should be stated here. The name(s) of the study organiser(s) and institute
should be stated under Referee (section 14). (Calibri 14)
This NMKL method has not been validated in an interlaboratory study.
If an interlaboratory study has not been carried out, this section should state
that and which validation that has been elaborated. (Calibri 14)

NMKL Method No. xxx, Year 20xx, Ed. xx Page 1 /

1. INTRODUCTION

(The text is to be written in Calibri font size 11)

The introduction is used to present additional information, such as comments concerning the
technical content of the method or the reasons for its preparation. Background information should
be included in this section.

2. ADDITIONS

Describe any major changes made in the method compared to the previous edition of the method.
Furthermore, other information, which does not fit within any of the other sections, can be
provided.

3. SCOPE AND FIELD OF APPLICATION

This section shall state:

 concisely what is analysed

 products/matrices to which the method applies
 possible limitations
 detection limit and/or quantification limit
 additional, useful information which could not be included in the title
 possible interferences

It shall also contain sufficient information to enable the user to judge quickly whether the method is
applicable to the products being considered, or whether limitations exist.

It is sometimes necessary to provide several procedure descriptions for the determination of a given
entity, depending, for example, on the composition of the product or on its differing contents for
that entity, or the accuracy required, each procedure having its own scope and field of application. A
clear distinction shall be drawn between the individual scopes of the procedures laid down if the
method includes several procedures.

4. DEFINITION

4. ALL CAPS, FROM THE EXTENDED FONT MENU (CTRL+D) (CALIBRI 12)
Text (Calibri 11)
4.1. Bold (Calibri 12)
Text (Calibri 11)
4.1.1. Bold (Calibri 11)
Text (Calibri 11)
This section shall give any definitions of terms used in the text that may be necessary for its
complete understanding. It is recommended that reference be made to already existing definitions
and/or terminology wherever possible. If numbering is required, please use a list or multilevel list
following the formatting in the example above.

5. REFERENCES

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Refer to the documents, indicating either the most recent edition or year or as undated; only stating
the number. If the reference is specified without the year of publication, it might be noted that it
refers to the latest version of the document, inc. possibly additions.

Regarding sampling, refer to NMKL Procedure No 12: Guide on sampling for analysis of foods.

The following documents should often be referred to in microbiological methods:

- NMKL Procedure No. 23: Guide on quality assurance in microbiological laboratories

- NMKL Method No. 91: Preparation of the test sample and initial suspension of food and
animal feeding stuffs for quantitative microbiological examination

6. PRINCIPLE

This section indicates the essential steps in the method used, the basic principles and the properties
of which use is made and, if appropriate, the reasons justifying the choice of certain procedures.

The text should be in accordance with the text of section 3, “Definitions” in terms of the properties
tested.

Chemical reactions can be included in this section if they are considered necessary for the
comprehension of the text or the calculations. These reactions shall, if appropriate, be expressed in
ionic form. When titrations are involved, the reactions are particularly useful in indicating the
number of equivalents in each mole of reactant.

7. SAFETY PRECAUTION

This section specifies general safety instructions.

8. REAGENTS AND PREPARATION OF SOLVENTS/ CULTURE MEDIA AND REAGENTS

The title of this section and subsections are selected based on what is appropriate for the respective
method.

FOR CHEMISTRY / SENSORY METHODS:

Water shall meet the requirements specified in NMKL Procedure No 23. In the method description
use the term "distilled or equivalent water”.

8.1 List of chemicals

The section shall begin, if applicable, with the following sentence or a suitably modified version of it:
“Use reagents of recognized analytical grade and distilled or demineralized water or water of
equivalent purity.”

This section shall list all the reagents and materials used during the test, together with their essential
characteristics (concentration, density, etc.), and shall specify, if necessary, their degree of purity. If
they exist, Chemical Abstract Service Registry numbers (CAS numbers) should preferably be given.
The list shall not include products used only in the preparation of a reagent unless the products are
used several times.

NMKL Method No. xxx, Year 20xx, Ed. xx Page 3 /

The reagents and materials shall be identified by a (sequential) reference number. These numbers
are referred to (in parentheses) in the procedure description. It is not necessary, however, to repeat
the reference number every time if no ambiguity is introduced.

If necessary, the precautions to be taken for storing the reagents, and the time for which they may
be stored, shall be specified.

It is desirable to list the substances/reagents in the following order:

 products used in their commercially available form

 aqueous solutions
 solutions of defined concentration
 standard reference solution (in mol/L)
 standard solution (in g/L(kg) or a submultiple thereof).

The expression “V1 +V2” means that the volume V1 of the solution in question is/has been added to
the volume V2 of the solvent.

Expressions such as “V1:V2” or “V1/V2” which have different meanings in different countries shall
not be used.

8.2 Preparation of solvents

8.3 Standards

8.4 Standard solutions

FOR MICROBIOLOGY METHODS:

Water shall meet the requirements specified in NMKL Procedure No 23. In the method description
use the term "distilled or equivalent water”.

The possibility of using commercial substrates include, for example by reading "Commercial
dehydrated culture media / materials are available and can be advantageously used. Using these,
follow the manufacturer's instructions. "

Where relevant include safety precautions in connection with the preparation.

Sterilization conditions should be stated.

8.1 Diluent
Diluent and its composition must always be stated.

pH tolerances of diluent and substrates must be consistent with NMKL Procedure No. 23, i.e. ± 0.2,
unless there is scientific justification for more stringent requirements.

The pH of the diluent and the substrates relates to the substrate ready for use, measured at 20 to 25
º C. Include in the description: "After sterilization (heat treatment), the pH of the ready for use
medium (dilution) should be x.y ± 0.2 measured at 20 to 25 ºC." The time of the pH measurement is
not conclusive and shall not be specified in the method.

NMKL Method No. xxx, Year 20xx, Ed. xx Page 4 /

The choice of diluent should be in accordance with NMKL Method No. 91, unless there are technical
reasons for another composition.

8.2 Liquid enrichment broths, substrates and reagents for confirmation

8.3.1 Name of broth, substrate or reagent

The international name shall appear in the English text.

In the case of substrates with mixtures, the first sub-paragraph describes the composition and
preparation of the basis substrate and the following sub point(s) describes the composition and
preparation of the mixture(s).

All the ingredients of substrates are to be included, including the chemical formula, and any crystal
water if relevant.

If it is essential for the quality of the substrate, the ingredients must be stated with the appropriate
number of decimals, also for the whole gram and ml.

The water in substrates is given as the amount of water to be added. Where possible the
composition of the substrate is given for the addition of 1000 ml of water. It should be clear stated if
addition of water for dilutions, substrates are to a certain quantity, e.g. 1000 ml, or if an exact
quantity is to be added (e.g. add 1000 ml).

8.3 Solid plating and confirmation media and reagents

8.3.1 Name of medium or reagent

The thickness of the plates or the amount of the substrates should be indicated, for example by the
following text: "Pour the melted substrate in Petri dishes to a thickness of 3 mm. For a Petri dish
with a diameter of 90 mm usually 18-20 ml agar required, or as specified in the method description.
If the plates should be incubated beyond 48 hours or at temperatures above 40 °C, it is necessary
with more agar."

Requirements for equivalent substrates

The general requirement of equivalent substrates is that they contain the same ingredients, both
qualitatively and quantitatively as given in the method, and that physical properties such as pH are
also identical.

A determination on whether substrates of different composition and/or other characteristics are

equal must depend on a concrete assessment in each case.

9. APPARATUS AND EQUIPMENT

This section shall list the names and significant characteristics of all the apparatus and equipment,
other than ordinary laboratory apparatus, to be used.

The items of apparatus shall be identified by a (sequential) reference number and referred to in the
procedure description. It is not necessary, however, to repeat this reference number every time if no
ambiguity is introduced.

10. SAMPLING

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Refer as appropriate to specific sampling methods or to NMKL Procedure No. 12

The following terminology should be used.

 Laboratory sample
 Test sample
 Test portion

11. PROCEDURE

The contents in this section will differ depending on whether this is a microbiology method, a
chemistry method or a sensory method and is outlined accordingly. The section may be divided into
subsections as required, exemplified below.

FOR CHEMISTRY METHODS:

The procedure for the method developed according to NMKL protocols 1, 5 and 6 should be
described (https://www.nmkl.org/resources/method-development/):

 NMKL Protocol No. 1: Referentvejledning for det kemiske område – Udarbejdelse af

analysemetoder indenfor NMKL (Danish only)
 NMKL Protocol No. 5: Analytical Quality Control – Guidelines for the publication of analytical
results of chemical analyses in foodstuffs. This protocol gives a guideline for publications on
what that should to be included and reviewed when elaborating, review or issuing articles
where chemical results are included.
 NMKL Protocol No. 6: Requirements for validation of chemical NMKL methods that are not
to be validated collaboratively

11.1 Pre-treatment
This subsection shall give all the information necessary for the preparation of the test portion from
the test sample or from the laboratory sample. It shall state the mass or volume of the test portion,
the accuracy with which this must be measured and, if necessary, any other relevant characteristics.

11.2 “Name of the procedure to be performed”

11.2.1 Name of specific sub-procedure or technique

11.3 …

FOR SENSORY METHODS:

11.1 Pre-treatment
This subsection shall give all the information necessary for the preparation of the test portion from
the test sample or from the laboratory sample. It shall state the mass or volume of the test portion,
the accuracy with which this must be measured and, if necessary, any other relevant characteristics.

11.2 “Name of the procedure to be performed”

NMKL Method No. xxx, Year 20xx, Ed. xx Page 6 /

11.2.1 Name of specific sub-procedure or technique

11.3 …

FOR MICROBIOLOGY METHODS

The procedure for the method developed according to NMKL protocols 3 and 7 should be described
(https://www.nmkl.org/resources/method-development/):

 NMKL Protocol No. 3: Guide for referees within microbiology. Elaboration of analytical
methods in NMKL
 NMKL Protocol No. 7: Requirement to internal (inhouse/single laboratory) validation of
presumptive NMKL microbiological methods

For quantitative methods:

11.1 Pre-treatment and dilution

This subsection shall give all the information necessary for the preparation of the test portion from
the test sample or from the laboratory sample. It shall state the mass or volume of the test portion,
the accuracy with which this must be measured and, if necessary, any other relevant characteristics.

Microbiological methods should refer to NMKL Method No. 91.

11.2 Plating

11.3 Incubation
Temperature tolerances: ± 1.0 °C for air incubators and ± 0.2 °C for water incubators, unless there
are scientific reasons for stricter requirements. Temperature settings are given as: 37.0 ± 1.0 °C.

Generally, the following temperatures are applied: 20, 25, 30, 37, 42 and 44 °C. Other incubation
temperatures are used only if scientifically reasoned.

Common incubation times:

 24 hours ± 3 hours
 48 hours ± 4 hours
 72 hours ± 6 hours

11.3 Reading of plates

Perform the reading, calculations and the stating of the results in accordance with the NMKL
Procedure No. 23.

When applicable, state a possible colony interval for reading.

For qualitative methods the results are given as detected / not detected in the amount of the sample
analysed.

11.4 Confirmation

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Use the following statement, unless otherwise scientifically reasoned: "Test a selection of suspect/
presumptive colonies - usually five of each type - if confirmation is necessary or desirable."

For qualitative methods:

11.1 Pre-treatment/ Test portion and initial suspension

This subsection shall give all the information necessary for the preparation of the test portion from
the test sample or from the laboratory sample. It shall state the mass or volume of the test portion,
the accuracy with which this must be measured and, if necessary, any other relevant characteristics.

Microbiological methods should refer to NMKL Method No. 91.

11.2 (pre-Enrichment)

11.2 (Selective) Enrichment

11.2 Plating
This title may vary and if treatments or plating steps before this title apply, they are presented in
sequential order as separate titles.

11.3 Incubation
Temperature tolerances: ± 1.0 °C for air incubators and ± 0.2 °C for water incubators, unless there
are scientific reasons for stricter requirements. Temperature settings are given as: 37.0 ± 1.0 °C.

Generally, the following temperatures are applied: 20, 25, 30, 37, 42 and 44 °C. Other incubation
temperatures are used only if scientifically reasoned.

Common incubation times:

 24 hours ± 3 hours
 48 hours ± 4 hours
 72 hours ± 6 hours

11.4 Reading of plates

Perform the reading, calculations and the stating of the results in accordance with the NMKL
Procedure No. 23.

When applicable, state a possible colony interval for reading.

For qualitative methods the results are given as detected / not detected in the amount of the sample
analysed.

11.4 Confirmation
Use the following statement, unless otherwise scientifically reasoned: "Test a selection of suspect/
presumptive colonies - usually five of each type - if confirmation is necessary or desirable."

NMKL Method No. xxx, Year 20xx, Ed. xx Page 8 /

12. CALCULATION OF RESULTS

Include how the calculations are to be carried out, the number of significant figures, as well as the
unit(s) to be stated.

13. EXPRESSION OF THE RESULT

State how the results should be expressed, which unit etc.

14. RELIABILITY OF THE METHOD

This section specifies the year of the collaborative study, the matrixes and levels, the number of
participating laboratories and the organiser of the study.

For qualitative methods, include the specificity and the sensitivity.

For quantitative methods include the precision (repeatability and reproducibility). The results from
the study should be given in a schematic form, as an annex.

For methods not validated in interlaboratory study state: “This NMKL method has not been validated
in an interlaboratory study.“

©NMKL. NMKL is responsible for the reliability of the method and questions
should be directed to the NMKL secretariat: post@nmkl.org.

NMKL Method No. xxx, Year 20xx, Ed. xx Page 9 /

ANNEX 1
Annexes can be included at the end of the method.

ANNEX 2

©NMKL

NMKL Method No. xxx, Year 20xx, Ed. xx Page 10 /