SIST-TS CEN/TS 16945:2016

SLOVENSKI STANDARD
SIST-TS CEN/TS 16945:2016
01-julij-2016
0ROHNXODUQHGLDJQRVWLþQHSUHLVNDYHLQYLWUR6SHFLILNDFLMH]DSUHGSUHLVNRYDOQH
SURFHVHPHWDERORPLNHYXULQXVHUXPXLQSOD]PLYHQVNHNUYL
Molecular in vitro diagnostic examinations - Specifications for pre-examination processes
for metabolomics in urine, venous blood serum and plasma
Molekularanalytische in-vitro-diagnostische Verfahren - Spezifikationen für
präanalytische Prozesse für Metabolomuntersuchungen in Urin, venösem Blutserum und
-plasma
Ta slovenski standard je istoveten z: CEN/TS 16945:2016
ICS:
11.100.10 'LDJQRVWLþQLSUHVNXVQL In vitro diagnostic test
VLVWHPLLQYLWUR systems
SIST-TS CEN/TS 16945:2016 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

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SIST-TS CEN/TS 16945:2016

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SIST-TS CEN/TS 16945:2016


CEN/TS 16945
TECHNICAL SPECIFICATION

SPÉCIFICATION TECHNIQUE

May 2016
TECHNISCHE SPEZIFIKATION
ICS 11.100.10
English Version

Molecular in vitro diagnostic examinations - Specifications
for pre-examination processes for metabolomics in urine,
venous blood serum and plasma
Tests de diagnostic moléculaire in vitro - Spécifications Molekularanalytische in-vitro-diagnostische Verfahren
relatives aux processus préanalytiques pour l'analyse - Spezifikationen für präanalytische Prozesse für
du métabolome dans l'urine et le sang veineux (sérum Metabolomuntersuchungen in Urin, venöses Blutserum
et plasma) und -plasma
This Technical Specification (CEN/TS) was approved by CEN on 22 March 2016 for provisional application.

The period of validity of this CEN/TS is limited initially to three years. After two years the members of CEN will be requested to
submit their comments, particularly on the question whether the CEN/TS can be converted into a European Standard.

CEN members are required to announce the existence of this CEN/TS in the same way as for an EN and to make the CEN/TS
available promptly at national level in an appropriate form. It is permissible to keep conflicting national standards in force (in
parallel to the CEN/TS) until the final decision about the possible conversion of the CEN/TS into an EN is reached.

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
© 2016 CEN All rights of exploitation in any form and by any means reserved Ref. No. CEN/TS 16945:2016 E
worldwide for CEN national Members.

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Contents Page
European foreword . 3
Introduction . 4
1 Scope . 5
2 Normative references . 5
3 Terms and definitions . 5
4 General Considerations. 7
5 Urine . 8
5.1 Outside the laboratory . 8
5.1.1 Urine collection manual . 8
5.1.2 Transport requirements. 9
5.2 Inside the laboratory . 9
5.2.1 Specimen reception . 9
5.2.2 Storage requirements . 9
5.2.3 Urine sample processing . 10
5.2.4 Long-term storage requirements for urine samples . 10
5.2.5 Urine thawing . 10
6 Blood . 10
6.1 Outside the laboratory . 10
6.1.1 Primary blood collection manual . 10
6.1.2 Transport of pre-processed specimens to laboratory . 12
6.2 Inside the laboratory . 12
6.2.1 Specimen reception . 12
6.2.2 Sample processing . 12
6.2.3 Transport of processed samples to a laboratory for metabolomics analysis or
transport to a biobank . 12
6.2.4 Long-term storage requirements . 13
6.2.5 Serum and plasma thawing and use . 13
1
Annex A (informative) Long-term stability of urine and serum H NMR metabolic profiles . 14
A.1 General . 14
1
A.2 Urine H NMR measurement result . 14
1
A.3 Serum H NMR measurement result . 16
A.4 NMR methods for urine and serum . 16
Bibliography . 18

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European foreword
This document (CEN/TS 16945:2016) has been prepared by Technical Committee CEN/TC 140 “In vitro
diagnostic and medical devices”, the secretariat of which is held by DIN.
Attention is drawn to the possibility that some of the elements of this document may be the subject of
patent rights. CEN 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 announce this Technical Specification: 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.
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Introduction
Molecular in vitro diagnostics has enabled a significant progress in medicine. Further progress is
expected by new technologies analysing signatures of nucleic acids, proteins, and metabolites in human
tissues and body fluids. However, the profiles of these molecules can change drastically during primary
sample collection, transport, storage, and processing thus introducing biases and making the outcome
from diagnostics or research unreliable or even impossible because the subsequent analytical assay will
not determine the situation in the patient but an artificial profile generated during the pre-examination
process. Therefore, a standardization of the entire process from sample collection to metabolomics
analysis is needed. Studies have been undertaken to determine the important influencing factors. This
Technical Specification draws upon such work to codify and standardize the steps for urine, serum and
plasma metabolomics analysis in what is referred to as the preanalytical phase.
Metabolomics, the global profiling of metabolites (namely molecules with a molecular weight
MW ≤ 2 000 Da [3]) in biological samples, is the determination of the dynamic multi-parametric
metabolic response of living systems to pathophysiological stimuli and/or genetic modification.
Metabolomics studies, which can be semiquantitative or quantitative, help in identifying metabolic
profiles that are characteristic for given pathological conditions, for disease prognosis, for the
evaluation of the individual response to medical intervention and pharmaceutical treatments.
Metabolites are physically and chemically different, and include e.g. sugars, acids, bases, and lipids [3].
This diversity of metabolites and the dynamic range of their concentration in biological samples
complicate the separation and detection methods and make it impossible to identify all the metabolites
in a single experiment. However, new high-throughput technologies based on NMR (nuclear magnetic
resonance) spectroscopy and MS (mass spectrometry) hold great potential due to their ability to look at
large parts of the whole metabolome, although with different sensitivity. These two main analytical
platforms are now well standardized. Equally well established are the statistical approaches needed to
extract information from the huge amount of data resulting from metabolomic analysis.
The metabolic profiles are very sensitive to preanalytical variations that can result from enzymatic
activity in the samples and chemical reactions (e.g. oxidation, [4], [5]). This Technical Specification
series provides guidelines arising from systematic studies conducted on the most commonly employed
biofluids: urine and blood derivatives, serum and plasma.
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1 Scope
This Technical Specification covers the preanalytical phase and recommends the handling,
documentation and processing of urine, venous blood plasma and serum intended for metabolomics
analysis. This Technical Specification is applicable to metabolomics examinations and is of importance
to biomedical laboratories, customers of laboratories, in vitro diagnostics developers and
manufacturers, institutions and companies performing biomedical research, biobanks, and regulatory
authorities.
The adoption of the described procedures for the preanalytical phase make it possible to compare and
evaluate the results obtained from metabolic profiling analysis.
2 Normative references
The following documents, in whole or in part, are normatively referenced in this document and are
indispensable for its application. For dated references, only the edition cited applies. For undated
references, the latest edition of the referenced document (including any amendments) applies.
EN ISO 15189:2012, Medical laboratories - Requirements for quality and competence (ISO 15189:2012,
Corrected version 2014-08-15)
ISO 15190, Medical laboratories — Requirements for safety
3 Terms and definitions
For the purposes of this document, the terms and definitions given in EN ISO 15189:2012 and the
following apply.
3.1
analytical phase
processes that start with the isolated analyte and include all kind of parameter testing or chemical
manipulation for quantitative or qualitative analysis
Note 1 to entry: For metabolomic analysis, analyte isolation is not necessarily required.
3.2
biofluid
biological fluid which can be excreted (such as urine or sweat), secreted (such as breast milk, saliva or
bile), obtained with a needle (such as blood or cerebrospinal fluid), or produced as a result of a
pathological process (such as blister or cyst fluid)
3.3
fasting
abstinence from any solid or liquid food excluding water
3.4
mass spectrometry
MS
method used to analyse chemical compounds on the basis of their mass to charge ratio
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3.5
metabolic profiling
use of analytical platforms to simultaneously measure the ensemble of metabolites that are accessible
to the employed (or selected) technique
EXAMPLE Examples for such techniques are NMR and MS.
3.6
metabolites
small molecules (≤ 2000 Da) that are intermediates and/or products of metabolism
Note 1 to entry: For further information see [3].
3.7
metabolome
complete set of metabolites to be found within an organism or a biological sample
Note 1 to entry: For further information see [3].
3.8
metabolomics
scientific study of the whole metabolome present within a biological sample (e.g., organism, cell, tissue
or biofluids) under a given set of conditions
3.9
MS-based metabolomics
use of mass spectrometry to measure metabolites in biological samples
3.10
Nuclear magnetic resonance spectroscopy
NMR
method where the resonance magnetic properties of atomic nuclei are used to determine physical and
chemical properties of atoms and molecules
[SOURCE: ISO/TS 80004-6:2013, 4.26]
3.11
NMR-based metabolomics
use of NMR spectroscopy to measure metabolites in biological samples
3.12
plasma
liquid part of unclotted blood
Note 1 to entry: Plasma samples can contain anti-coagulants.
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3.13
pre-examination processes
preanalytical phase
preanalytical workflow
processes that start, in chronological order, from the clinician’s request and include the examination
request, preparation and identification of the patient, collection of the primary sample(s), temporary
storage, transportation to and within the analytical laboratory, aliquotting, retrieval, isolation of
analytes, and end when the analytical examination begins
Note 1 to entry: The preanalytical phase can include preparative processes that can influence the outcome of
the intended examination.
[SOURCE: EN ISO 15189:2012, 3.15, modified — An additional term was added and more details were
included.]
3.14
primary sample
specimen
discrete portion of a body fluid, breath, hair or tissue taken for examination, study or analysis of one or
more quantities or properties assumed to apply for the whole
[SOURCE: EN ISO 15189:2012, 3.16, modified — The term and definition are used here without the
original notes.]
3.15
room temperature
temperature which is defined as 18 °C to 25 °C for the purpose of this document
3.16
serum
liquid that can be separated from clotted blood
3.17
stability
ability of a sample material, when stored under specified conditions, to maintain a stated property value
within specified limits for a specified period of time
Note 1 to entry: The analytes for the purpose of this document are metabolites.
[SOURCE: ISO Guide 30:1992, 2.7]
4 General Considerations
For general statements on specimen collection and handling (including avoidance of cross
contaminations) see EN ISO 15189:2012, 5.2.6, 5.4.4. Consumables including kits shall be verified
before use in examination (see EN ISO 15189:2012, 5.3.2.3); EN ISO 15189:2012, 5.5.1.2 and 5.5.1.3 can
also apply.
As all steps of a diagnostic workflow can influence the final analytical performance, the entire workflow
comprising the preanalytical steps, including information on specimen stability and storage conditions,
and analytical steps should be verified and validated (see EN ISO 15189).
In the absence of suitable specimen stabilization technologies, regarding the metabolome, the specimen
collection should be carried out in hospital premises or institutions where there are immediate suitable
biofluid processing procedures available.
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Specifically for specimens intended to be analysed by metabolomics, the following steps shall be
considered:
a) the specimen collection from the patient;
b) the selection of collection containers and packages (e.g. cooling box, box for storing and
transportation);
c) the selection of stabilization procedures (e.g. any compounds added for stabilizing the specimen);
d) the recording of any additions or modifications to the specimen;
e) the recording of types and quantity and description of specimens.
Safety regulations on facilities, transport and handling shall be considered (see EN ISO 15189:2012,
5.2.3 and 5.4.5, and ISO 15190).
5 Urine
5.1 Outside the laboratory
5.1.1 Urine collection manual
5.1.1.1 Information on the primary specimen donor
The documentation should include, but is not limited to:
a) the specimen donor/patient ID, which can be in the form of a code;
b) the health status and relevant lifestyle factors of the urine donor (e.g. healthy, disease type, diet,
gender, age);
c) the information about medical treatment and special treatment prior to urine collection (e.g.
anaesthetics, medications);
d) the collection time, including information about fasting, previous activities.
See also EN ISO 15189:2012, 5.4.4.
5.1.1.2 Selection and labelling of collection containers
The laboratory shall define the container intended for urine collection.
Additives are usually not used, because they can interfere with the analytical method. If they are
required, their impact on the analytical performance and outcome shall be analysed. Additives can be
harmful (e.g. toxic or corrosive).
A sufficient minimum volume of urine should be collected according to the requirements of the
preanalytical preparation steps and the analytical test. For the labelling (specimen identification) of the
urine collection tube a routine procedure (EN ISO 15189:2012, 5.4.4.3, e)) or a procedure with
additional information (e.g. 2D-barcode) shall be used.
5.1.1.3 Urine collection and reception from the specimen donor
Instruction for the urine collection shall be given to the donor, including any safety measures
concerning additives in the collection container.
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The first midstream urine of the morning should be collected after a minimum of 8 h fasting. Drinking
can influence urine metabolite concentrations. This requires a normalization. Specify, if collected at
different times, or for 24-h collection. Any variations to standard instructions shall be validated.
NOTE This enables to perform the metabolomics analysis of urine where donors are synchronized having
similar metabolic conditions. Research or dedicated analytical tests can require different patient conditions.
Any clinical procedure affecting the specimen collection shall be documented. The total volume to be
collected shall be documented
The identity of the person receiving the specimen from the patient and the time of urine collection
according to EN ISO 15189, 5.4.4.3, f) shall be documented.
5.1.1.4 Information on the urine specimen and storage requirements at the urine collection site
As metabolic profiles can change after urine collection and can thereby affect the validity and reliability
of the analytical test result, the documentation on the primary urine specimen shall include the time
and date of urine collection.
The whole urine specimen should be kept refrigerated at 2 °C to 8 °C for a maximum of 2 h and shall not
be frozen prior to
...