A011 Guidelines for checking and validating test and calibration methods according to ISO/IEC 17025

01.09.2008 Version 04 Page 1 de 7

A011 Guidelines for checking and validating test and calibration methods according to ISO/IEC 17025

Modifications: document completely updated

South Lane Tower I 1, avenue du Swing L-4367 Belvaux Tl.: (+352) 2477 4360 Fax: (+352) 2479 4360 olas@ilnas.etat.lu www.ilnas.lu

Checked by Jean-Marie Reiff

Approved by Dominique Ferrand

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A011 Guidelines for checking and validating test and calibration methods according to ISO/IEC 17025
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CONTENTS 1. Aims 2. Methodology 3. Summary of requirements 4. Definitions 5. Quantitative methods

5.1 Checking a standard method 5.2 Method validation

6. Qualitative methods
6.1 Checking a standard method 6.2 Method validation

7. Methods validated by an independent body 8. Documentation

8.1 Methods 8.2 Validation files

9. Bibliography

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A011 Guidelines for checking and validating test and calibration methods according to ISO/IEC 17025
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1. Aim
These guidelines set out the items that audited laboratories must provide as part of an accreditation audit to meet the requirements of standard ISO/IEC 17025 relating to the checking and validation of test and calibration methods submitted for accreditation. When laboratories make use of test and calibration methods developed in-house, their validation is an essential step. To strengthen the implementation of this practice, OLAS accreditation is only granted on condition that the validation stage has been completed at the time of the audit for obtaining accreditation.

2. Methodology
These guidelines have been compiled on the basis of international or national standards and of various guidelines drawn up by accreditation bodies, members of the EA, a list of which is attached to the bibliography. A technical training day was organised on the 1st September 2005 at the premises of OLAS, which was attended by the heads of 10 accredited Luxemburg laboratories or accreditation candidates. This day was run by 3 experts and made it possible to gather the opinions of chief technicians and throw light on certain queries concerning method validation. Following this technical training day OLAS was asked to draw up these guidelines for which a plan was circulated amongst the participants in December 2005. This guide was then disseminated by including the comments and suggestions accepted by OLAS.

3. Summary of requirements
Standard ISO/IEC 17025 stipulates that: The laboratory must use test and/or calibration methods which meet the clients needs and which are suitable to the tests and/or calibrations carried out, preferably using methods published as international, regional or national standards. The laboratory may equally employ methods developed in-house or methods it has adopted provided these are suitable for the expected purpose and that they have been validated. Laboratories must validate non-standard methods, methods designed and developed inhouse, methods used outside their expected application, including the extension and modification of standard methods, to confirm that the methods are suitable for the expected purpose. Validation must be as comprehensive as necessary to meet the needs of the application or the given field of application. The laboratory must record results obtained, the validation operating procedure, as well as a declaration of the methods suitability to the expected use. The laboratory must confirm that it can correctly apply the standard methods before implementing them for tests and calibrations. From these ISO/IEC 17025 requirements we may deduce that a standard method used by the laboratory must be checked to confirm its correct application by the laboratory and, if it is not a standard method, it must be validated. It is the responsibility of the technical auditors to define within their area of competence whether the method is standard or not. Indeed, these (methods) may also be defined in regulations, or even in international and scientific journals, which can give them the status of reference standard. Equally the laboratory must document possible deviations in application from the standard method and prove that these deviations do not alter method performance. Where this is not the case, the method will not be considered standard.

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4. Definitions
Please find below a glossary of definitions relating to the subject: Accuracy Closeness of agreement between the mean value obtained from a series of test results and the value of an accepted reference. (ISO 5725-1:1994). Precision Closeness of agreement between independent test results obtained under stipulated conditions (ISO 5725-1:1994). Repeatability Precision under conditions of repeatability (ISO 5725-1:1994). Conditions of repeatability Conditions under which independent test results are obtained by the same method, on identical test samples in the same laboratory, by the same operator, using the same equipment, within a short period of time (ISO 5725-1:1994). Reproducibility Precision under conditions of reproducibility (ISO 5725-1:1994). Conditions of reproducibility Conditions under which test results are obtained by the same method on identical test samples in different laboratories, by different operators, using different equipment (ISO 5725-1:1994). When reproducibility is limited to the laboratory, it is known as inter and intra laboratory reproducibility. Linearity The ability of an analytical method to give the laboratory (within a defined range) an actual value or results proportional to the dose of analyte being measured in the sample (AFNOR XP T90-210 December 1999). Limits of linearity The limits of linearity are the limiting experimental values between which a linear calibration curve may be drawn with a known confidence level (AFNOR XP T90-210 December 1999). The range between the limits of linearity is the linear field. Specificity The degree to which an analytical method is exclusively suited to determine the amount of analyte under consideration ensuring that the response measured only derives from the analyte (AFNOR XP T90-210 December 1999). Blank Test done in the absence of a matrix (reactive blank) or on a matrix which does not contain any analyte (matrix blank) (AFNOR XP T90-210 December 1999). Limit of detection The smallest amount of the analyte to be tested in a sample, which may be detected and distinguished from the blank value (for a given probability) but may not necessarily be quantified (AFNOR XP T90210 December 1999). Limit of quantification The smallest amount of the analyte to be tested in a sample which may be determined quantitatively under the experimental conditions described in the method with a specified variation (AFNOR XP T90210 December 1999).
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A011 Guidelines for checking and validating test and calibration methods according to ISO/IEC 17025
01.09.2008 False positive (qualitative method) Positive result when the reference method gives a negative result. False negative (qualitative method) Negative result when the reference method gives a positive result. Standard method check A check by the laboratory of its ability to implement a standard method. Method validation Validation is the confirmation that the specific requirements of an expected application are met through investigation and through providing objective evidence (ISO/IEC 17025:2005). Method validation is generally internal, that is to say, carried out within the laboratory. Nevertheless, a move towards external method validation, that is to say, carried out by a qualified third party is observed. Version 04 Page 5 de 7

5. Quantitative methods
5.1 Standard method check
This involves determining the characteristics of a standard method as applied within the laboratory and checking, if needs be, that these characteristics are consistent with those defined by the standard method or with those determined by the Chief Laboratory Technician. The characteristics to be checked are:

repeatability, internal reproducibility, linear range (where applicable), limit of detection (where applicable), limit of quantification (where applicable).

Participation in inter laboratory tests, if they exist, is vital to ensure mastery of the method over time.

5.2 Method validation

Validation of a non-standard method must include the check-list items mentioned above to which are added determination of method specificity, and its accuracy by comparison to a reference method or to certified reference materials or solutions. The aims of method performance must also be determined beforehand by the Chief Laboratory Technician and checked during validation. Therefore the characteristics to be checked are: repeatability, internal reproducibility, linear range, limit of detection (where applicable), limit of quantification (where applicable). specificity, accuracy. Participation in inter laboratory tests, if they exist, is vital to ensure mastery of the method over time.

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A011 Guidelines for checking and validating test and calibration methods according to ISO/IEC 17025
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6. Qualitative methods
6.1 Standard method check
In the case of qualitative methods, the laboratory must, more so than in any other case, use standard methods because validation is complicated by the scarce availability of samples of known analyte concentration. Method checking relies on confidence in staff proficiency and follow-through as well as the results of routine participation in inter laboratory test programmes.

6.2 Method validation

If the laboratory uses non-standard methods they must, in addition, check the robustness of their method by determining its specificity, its detection limit (sensitivity) and its rate of false negatives and positives. That is to say the following parameters: specificity, its detection limit (sensitivity), false positives, false negatives, relative accuracy (if needs be). This involves determining the characteristics of the method such as it is used within the laboratory to check, if needs be, that these characteristics are consistent with those defined by the standard method or with those determined beforehand by the Chief Laboratory Technician or the client. They may rely on a reference, which if possible, should be independent of the equipment and reagent supplier, and must do their best to proceed with on-site experimental controls. Participating in inter laboratory testing and monitoring staff proficiency are of course vital.

7. Methods validated by an independent body

The laboratory may use methods validated by an independent body (where possible accredited), according to the requirements of ISO 16140 standard or an equivalent protocol. These methods are thus categorised as standard methods with regard to the requirements of ISO/IEC 17025 standard

8. Documentation
8.1 Methods
Methods that are implemented must be documented. Items of information that should be provided in the relevant document are: a) name and number of norms, standards, internal methods cited in reference, as well as the year of their last revision; b) area of application: products, types of samples, properties measured, range and limits of measurements; c) documentary references: list of supplementary documents which the method requires for its application; d) terminology: definition of specific terms, explanation of symbols and abbreviations used in the text; e) principles of the test: abstract or summary of the method;

Degree of conformity between the result obtained with the reference method and the result obtained with the alternative method on identical samples. The updated version of this document is available on www.ilnas.lu. The printed versions are not managed.

A011 Guidelines for checking and validating test and calibration methods according to ISO/IEC 17025
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apparatus: necessary equipment and instrument specifications;

g) reagents and other consumable products, where applicable; h) samples: number and amount of sample or specimen required for the tests and the preparation method; i) j) preparation: calibration or check, instrument packaging, and possible environmental requirements; operating method: procedure for carrying out the test;

k) calculation and interpretation of results: to include, where possible, a means of calculating the confidence limits of the results; l) method performance characteristics determined during the monitoring or validation phase;

m) quality control provisions.

8.2 Validation files

Items which lead to method checking or validation must be kept together in a file available to the technical auditors who may request it before the laboratory audit or examine it during the audit.

9. Bibliography
1. ISO/IEC 17025:2005: General recommendations regarding the proficiency of test and calibration laboratories. 2. ISO 5725: Accuracy (accuracy and precision) of measurement methods and results Part-1:1994(E): General principles and definitions Part-2:1994(E): Basic method for the determination of the repeatability and reproducibility of a standard measurement method. Part-4:1994(E): Basic method for the determination of the accuracy of a standard measurement method 3. ISO 11843:1997: Capability of detection Part 1: Terms and definitions, Part 2: Methodology in linear calibration. 4. ISO 3534-1:1993 (E/F): Statistics Vocabulary and symbols Part 1: Probabilities and general statistical terms 5. ENV ISO 13843: Water quality Guidelines for the validation of microbiology methods (ISO/TR 13843:2000). 6. Thompson, M., Ellison, S.L.R. and Wood, R.: Harmonised Guidelines for Single-Laboratory Validation of Methods of Analysis, Pure Appl. Chem., 74, 835-855 (2002). 7. Kromidas, S.: Handbook of Validation in Analysis, Verlag Wiley-VCH, Weinheim, ISBN 3-52729811-8 (2000). 8. Committee decision of 12th August 2002 regarding the methods of application of Council directive 96/23/EC regarding the performance of analytical methods and the interpretation of results. Official Journal of the European Community L221: 8-36 (2002). 9. XP T90-210 December 1999: Protocol for evaluating an alternative method of physico-chemical quantitative analysis compared to a reference method (AFNOR). 10. DIN NAW IW1, Strategies in Water Analysis, Routine Validation and Quality Assurance, Entwurf (1997). 11. Guidelines for the validation of chemico-physical test methods and assessment of uncertainty in measurement: SAS (Swiss Accreditation Society). Document 326f edition December 2004. 12. Guidelines for method validation in medical biology: COFRAC (French Committee for Accreditation). Document LAB GTA 04 June 2004.
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