Ich q2r2 Guideline Validation Analytical Procedures Step 5 Revision 1 en

14 December 2023
EMA/CHMP/ICH/82072/2006
Committee for Medicinal Products for Human Use
ICH Q2(R2) Guideline on validation of analytical

procedures
Step 5
Transmission to CHMP 8 March 2022
Adoption by CHMP 24 March 2022
Release for public consultation 31 March 2022
Deadline for comments 31 July 2022
Final adoption by CHMP 14 December 2023
Date for coming into effect 14 June 2024
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Document History
Code History Date
Q2 Approval by the Steering Committee under Step 2 and 26 October 1993

release for public consultation.
Q2A Approval by the Steering Committee under Step 4 and 27 October 1994
recommendation for adoption to the three ICH regulatory
bodies.
Q2B Approval by the Steering Committee under Step 2 and 29 November 1995
release for public consultation.
Q2B Approval by the Steering Committee under Step 4 and 6 November 1996
recommendation for adoption to the three ICH regulatory
bodies.
Q2(R1) The parent guideline is now renamed Q2(R1) as the November 2005
guideline Q2B on methodology has been incorporated to
the parent guideline. The new title is “Validation of
Analytical Procedures: Text and Methodology”.
Q2(R2) Complete revision of guideline to include more recent 24 March 2022

application of analytical procedures and to align content
with Q14.
Endorsement by the Members of the ICH Assembly under

Step 2 and release for public consultation.
Q2(R2) Adoption by the Regulatory Members of the ICH Assembly 1 November 2023
under Step 4.
Q2(R2) Error Correction to Table 5: Dissolution with HPLC as 30 November 2023

product performance test for an immediate release dosage
form Reportable Range Linearity formulae on page 25;
Correction to Tables 6-11 on pages 26-32.
EMA/CHMP/ICH/82072/2006 Page 2/33

ICH Q2(R2) Guideline on validation of analytical
procedures
Table of contents
1. Introduction ........................................................................................................ 4
1.1 Objective ............................................................................................................... 4
1.2 Scope.................................................................................................................... 4
2. General considerations for analytical procedure validation ............................... 4
2.1 Analytical procedure validation study ........................................................................ 5
2.2 Validation during the lifecycle of an analytical procedure.............................................. 7
2.3 Reportable range .................................................................................................... 8
2.4 Demonstration of stability-indicating properties .......................................................... 8
2.5 Considerations for multivariate analytical procedures .................................................. 8
3. Validation tests, methodology and evaluation ................................................... 10
3.1 Specificity/ selectivity............................................................................................ 10
3.1.1 General considerations ................................................................................ 10
3.1.2 Recommended data .................................................................................... 10
3.2 Range ................................................................................................................. 11
3.2.1 General considerations ................................................................................ 11
3.2.2 Response................................................................................................... 12
3.3 Accuracy and precision .......................................................................................... 14
3.3.1 Accuracy ................................................................................................... 14
3.3.2 Precision ................................................................................................... 15
3.3.3 Combined approaches for accuracy and precision ........................................... 16
3.4 Robustness .......................................................................................................... 16
4. Glossary ............................................................................................................ 16
5. References......................................................................................................... 20
ANNEX 1: Selection of validation tests ........................................................................ 21
ANNEX 2: Illustrative examples for analytical techniques ........................................... 22

1. Introduction
1.1 Objective
This guideline presents elements for consideration during the validation of analytical procedures
included as part of registration applications. Analytical procedure validation forms a part of the
analytical procedure lifecycle, as described within ICH Q14 Analytical Procedure Development. ICH
Q2(R2) provides guidance on selection and evaluation of the various validation tests for analytical
procedures. This guideline includes a collection of terms and their definitions, which are meant to
bridge the differences that often exist between various compendia and documents of the ICH member
regulatory authorities.
The objective of validation of an analytical procedure is to demonstrate that the analytical procedure is
fit for the intended purpose. Further general guidance is provided on validation studies for analytical
procedures.
1.2 Scope
This guideline applies to analytical procedures used for release and stability testing of commercial drug
substances and products, hereafter referred to as ‘products’. The guideline can also be applied to other
analytical procedures used as part of the control strategy (ICH Q10 Pharmaceutical Quality System)
following a risk-based approach. The scientific principles described in this guideline can be applied in a
phase-appropriate manner to analytical procedures used during clinical development.
The guideline is directed to common uses of analytical procedures, such as assay, potency, purity,
impurity (quantitative or limit test), identity or other quantitative or qualitative measurements.
2. General considerations for analytical procedure

validation
This guideline indicates the data which should be presented in a regulatory submission. Analytical
procedure validation data should be submitted in the corresponding sections of the application (ICH
M4Q The Common Technical Document For The Registration Of Pharmaceuticals For Human Use).
Relevant data collected during validation (and any methodology used for calculating validation results)
should be submitted to demonstrate the suitability of the procedure for the intended purpose. Suitable
data derived from development studies (see ICH Q14) can be used as part of validation data. When an
established platform analytical procedure is used for a new purpose, validation testing can be
abbreviated, if scientifically justified.
Approaches other than those set forth in this guideline may be applicable and acceptable with
appropriate science-based justification. The applicant is responsible for designing the validation studies
and protocol most suitable for their product.
Reference materials, or other suitably characterised materials, with documented identity, purity, or any
other characteristics as necessary, should be used in the validation study.
In practice, the experimental work can be designed so that the appropriate performance characteristics
are considered simultaneously to provide sound, overall knowledge of the performance of the analytical
procedure, for instance: specificity/selectivity, accuracy, and precision over the reportable range.
As described in ICH Q14, the system suitability test (SST) is an integral part of analytical procedures
and is generally established during development as a regular check of performance. Robustness is
typically evaluated as part of development prior to the execution of the analytical procedure validation

study (ICH Q14). Finally, the analytical procedure validation strategy is developed based on knowledge
of the analytical procedure and the intended purpose. This includes the required analytical procedure
performance to ensure the quality of the measured result (ICH Q14). If successfully executed, the
analytical procedure validation strategy will demonstrate that the analytical procedure is fit for the
intended purpose.
2.1 Analytical procedure validation study
A validation study is designed to provide sufficient evidence that the analytical procedure meets its
objectives. These objectives are described with a suitable set of performance characteristics and
related performance criteria, which can vary depending on the intended purpose of the analytical
procedure and the specific technology selected. Section 3 “Validation tests, methodology and
evaluation” summarises the typical methodologies and validation tests that can be used (see also
Figure 2 in Annex 1 on selection of validation tests). Specific non-binding examples for common
techniques are given in Annex 2. Table 1 (below) provides the measured quality attributes, typical
performance characteristics and related validation tests, which are further illustrated in Annex 1.
The validation study should be documented. Prior to the validation study, a validation protocol should
be generated. The protocol should contain information about the intended purpose of the analytical
procedure, the performance characteristics to be validated and the associated criteria. In cases where
prior knowledge is used (e.g., from development or from previous studies), appropriate justification
should be provided. The results of the validation study should be summarised in a validation report.
The experimental design of the validation study should reflect the number of replicates used in routine
analysis to generate a reportable result. If justified, it may be acceptable to perform some validation
tests using a different number of replicates or to adjust the number of replicates in the analytical
procedure based on data generated during validation.
Figure 1 shows the inter-relationship between ICH Q2 and ICH Q14, and how knowledge generated
during analytical procedure development as described in ICH Q14 aids the design of a validation study.

Table 1: Typical performance characteristics and related validation tests for measured quality
attributes
Measured Quality IDENTITY IMPURITY (PURITY) ASSAY

Attribute
Other quantitative Content or potency
measurements (1)
Quantitative Limit Test

Other quantitative
Test
Analytical measurements (1)
Procedure
Performance
Characteristics to be
Demonstrated (2)
Specificity (3)
Specificity Test + + + +
Range
Response - + - +
(Calibration Model)
Lower Range Limit - QL† DL -
Accuracy (4)
Accuracy Test - + - +
Precision (4)
Repeatability Test - + - +
Intermediate - + (5) - + (5)

Precision Test
- signifies that this test is not normally conducted
+ signifies that this test is normally conducted
† in some complex cases DL may also be evaluated
QL, DL: quantitation limit, detection limit
(1) other quantitative measurements can follow the scheme for impurity, if the range limit is close to the DL/QL;
other quantitative measurements can follow the scheme for assay (content or potency), if the range limit is not
close to the DL/QL

(2) some performance characteristics can be substituted with technology-inherent justification in the case of certain
analytical procedures for physicochemical properties
(3) lack of specificity of one analytical procedure should be compensated by one or more other supporting analytical
procedures, unless appropriately justified
(4) alternatively, a combined approach can be used to evaluate accuracy and precision
(5) where reproducibility has been performed and intermediate precision can be derived from the reproducibility
data set, an independent study for intermediate precision is not required
Figure 1: Validation study design and evaluation
2.2 Validation during the lifecycle of an analytical procedure
Changes may be required during the lifecycle of a validated analytical procedure. In such cases, partial
or full revalidation may be required. Science and risk-based principles can be used to justify whether
or not a given performance characteristic needs revalidation. The extent of revalidation depends on the
performance characteristics impacted by the change.
Transfer of a validated analytical procedure should be considered in the context of analytical lifecycle
changes in line with ICH Q14. When transferring analytical procedures to a different laboratory, a
partial or full revalidation of the analytical procedure performance characteristics and/or comparative
analysis of representative samples should be performed. Justification for not performing additional
transfer experiments should be provided if appropriate.

Co-validation can be used to demonstrate that the analytical procedure meets predefined performance
criteria by using data generated at multiple sites and could also satisfy the requirements of analytical
procedure transfer at the participating sites.
2.3 Reportable range
The required reportable range is typically derived from the specification and depends on the intended
use of the procedure. The reportable range is confirmed by demonstrating that the analytical
procedure provides results with acceptable response, accuracy and precision. The reportable range
should be inclusive of the upper and lower specification or reporting limits, as applicable.
Table 2 exemplifies recommended reportable ranges for common uses of analytical procedures; other
ranges may be acceptable if justified. In some cases, e.g., at low amounts, wider upper ranges may be
more practical.
2.4 Demonstration of stability-indicating properties
A validated quantitative analytical procedure that can detect changes in relevant quality attributes of a
product during storage is considered to be stability-indicating. To demonstrate specificity/selectivity of
a stability-indicating test, samples containing relevant degradation products should be included in the
study. These can include: samples spiked with target analytes and known interferences; samples that
have been exposed to various physical and chemical stress conditions; and actual product samples that
are either aged or have been stored under stressed conditions.
2.5 Considerations for multivariate analytical procedures
For multivariate analytical procedures, results are determined through a multivariate calibration model
utilising more than one input variable (e.g., a spectrum with many wavelength variables). The
multivariate calibration model relates the input data to a value for the property of interest (i.e., the
model output).
Successful validation of a multivariate procedure should consider calibration, internal testing and
validation.
Typically, development and validation are performed in two phases.
 In the first phase, model development consists of calibration and internal testing. Calibration data
are used to create the calibration model. Test data are used for internal testing and optimisation of
the model. The test data could be a separate set of data or part of the calibration set used in a
rotational manner. This internal test step is used to obtain an estimate of the model performance
and to fine-tune an algorithm’s parameters (e.g., the number of latent variables for partial least
squares (PLS)) to select the most suitable model within a given set of data. For more details, see
ICH Q14.
 In the second phase, model validation, a validation set with independent samples is used for
validation of the model. For identification libraries, validation involves analysing samples (i.e.,
challenge samples) not represented in the library to demonstrate the discriminative ability of the
library model.

Table 2: Examples of reportable ranges for common uses of analytical procedures
Use of analytical Low end of reportable range High end of reportable range
procedure
Assay of a product (1) 80% of declared content or 80% 120% of declared content or
of lower specification acceptance 120% of the upper specification
criterion acceptance criterion
Potency Lowest specification acceptance Highest specification acceptance

criterion -20% criterion +20%
Content uniformity 70% of declared content 130% of declared content
Dissolution:
Immediate release
Q - 45% of the lowest strength
one point specification
Lower limit of reportable range 130% of declared content of the

(as justified by the specification) highest strength
multiple point specification
or QL, as appropriate.
Lower limit of reportable range

Modified release (as justified by the specification)
or QL, as appropriate.
Impurity (1) Reporting threshold 120% of specification acceptance

criterion
Purity (as area %) 80% of lower specification Upper specification acceptance

acceptance criterion criterion or 100%
(1) Where assay and impurity are performed as a single test and only one standard is used, linearity should be
demonstrated for both the reporting level of the impurities and up to 120% of the specification acceptance
criterion for assay.
Samples used for the validation of quantitative or qualitative multivariate procedures require values or
categories assigned to each sample, typically obtained by a reference analytical procedure, i.e., a
validated or pharmacopoeial procedure.
When a reference analytical procedure is used, its performance should equal or exceed the expected
performance of the multivariate analytical procedure. Analysis by the reference analytical procedure
and multivariate data collection should be performed on the same samples (whenever possible) within
a reasonable period of time to assure sample and measurement stability. In some cases, a correlation
or conversion may be needed to provide the same unit of measure. Any assumptions or calculations
should be described.

3. Validation tests, methodology and evaluation
In the following chapters, experimental methodologies to evaluate the performance of an analytical
procedure are described. These methodologies are grouped according to main performance
characteristics dictated by the analytical procedure design. It is acknowledged that information about
multiple performance characteristics may be derived from the same dataset. Different approaches may
be used to demonstrate that the analytical procedure meets the objectives and related performance
criteria, if justified.
3.1 Specificity/ selectivity
3.1.1 General considerations
The specificity or selectivity of an analytical procedure can be demonstrated through absence of

interference or comparison of results to an orthogonal procedure. In some cases, specificity/selectivity
may be inherently given by the underlying scientific principles of the analytical procedure. Some
experiments can be combined with accuracy studies.
Selectivity could be demonstrated when the analytical procedure is not specific. However, the test for
an analyte to be identified or quantitated in the presence of potential interference should minimise that
interference and demonstrate that the analytical procedure is fit for the intended purpose.
Where one analytical procedure does not provide sufficient discrimination, a combination of two or
more procedures is recommended to achieve the necessary specificity/selectivity.
3.1.1.1 Absence of interference

Specificity/selectivity can be shown by demonstrating that the identification and/or quantitation of an
analyte is not impacted by the presence of other substances (e.g., impurities, degradation products,
related substances, matrices, or other components likely to be present).
3.1.1.2 Orthogonal procedure comparison

Specificity/selectivity can be verified by demonstrating that the measured result of an analyte is
comparable to the measured result of a second, well characterised analytical procedure that ideally
applies a different measurement principle.
3.1.1.3 Technology inherent justification

In some cases where the specificity of the analytical technology can be ensured and predicted by
technical parameters (e.g., resolution of isotopes in mass spectrometry, chemical shifts in NMR
spectroscopy), additional experimental studies may not be required, if justified.
3.1.2 Recommended data
3.1.2.1 Identification
For identification tests, a critical aspect is to demonstrate the capability to identify the analyte of
interest based on unique aspects of its molecular structure and/or other specific properties. The
capability of an analytical procedure to identify an analyte can be confirmed by obtaining positive
results comparable to a reference material using samples containing the analyte, along with negative
results from samples which do not contain the analyte. In addition, the identification test should be
applied to materials structurally similar to or closely related to the analyte to confirm that a positive
result is not obtained. The choice of such potentially interfering materials should be based on scientific
judgement with a consideration of interferences that could occur.

3.1.2.2 Assay, purity and impurity test(s)
The specificity/selectivity of an analytical procedure should be demonstrated to fulfil the accuracy
requirements for the content or potency of an analyte in the sample.
Representative data (e.g., chromatograms, electropherograms, spectra, biological response) should be

used to demonstrate specificity and relevant components should be labelled, if appropriate.
For separation techniques, suitable discrimination should be investigated at an appropriate level (e.g.,
for critical separations in chromatography, specificity can be demonstrated by the resolution of the two
components which elute closest to each other). Alternatively, spectra of different components could be
compared to assess the possibility of interference.
For non-separation techniques (e.g., bioassay, ELISA, qPCR), specificity can be demonstrated through
the use of reference materials or other suitably characterised materials to confirm the absence of
interference in relation to the analyte. In cases where the analyte is a process-related impurity,
specificity (non-interference) must also be confirmed against the product.
In case a single procedure is not considered specific or sufficiently selective, an additional procedure
should be used to ensure adequate discrimination. For example, where a titration is used to assay a
drug substance for release, the combination of the assay and a suitable test for impurities may be
used.
Impurities or related substances are available or can be intentionally created:
For assay or potency, discrimination of the analyte in the presence of impurities and/or excipients
should be demonstrated. Practically, this can be performed by spiking product with appropriate
amounts of impurities and consequently demonstrating that the assay result is unaffected by the
presence of these materials (e.g., by comparison with the assay result obtained on unmanipulated
samples). Alternatively, samples containing appropriate amounts of impurities could be generated
through deliberate stressing of product materials.
For a purity or impurity test, discrimination can be established by stressing or spiking product to
achieve appropriate levels of impurities or related substances and demonstrating the absence of
interference.
Impurities or related substances are not available:
If impurities, related substances or degradation products cannot be prepared or isolated, specificity can
be demonstrated by comparing the test results of samples containing typical impurities, related
substances or degradation products with an orthogonal procedure. The approach taken should be
justified.
3.2 Range
3.2.1 General considerations
The range of an analytical procedure is the interval between the lowest and the highest results in
which the analytical procedure has a suitable level of response, accuracy and precision. The range can
be validated through the direct assessment of reportable results (to generate a reportable range) using
an appropriate calibration model (i.e., linear, non-linear, multivariate). In some cases, the reportable
range can be determined using one or more appropriate working ranges, depending on the sample
preparation (e.g., dilutions) and the analytical procedure selected.
Typically, a working range corresponds to the lowest and the highest sample concentrations or purity
levels presented to the analytical instrument for which the analytical procedure provides reliable

results. Mathematical calculations are typically required to generate reportable results. Reportable
range and working range could be identical.
In cases where materials of sufficient purity (or containing sufficient amounts of impurities) to validate
the full range (e.g., 100% purity) cannot be generated, extrapolation of the reportable range may be
appropriate and should be justified.
3.2.2 Response
3.2.2.1 Linear response

A linear relationship between analyte concentration and response should be evaluated across the range
of the analytical procedure to confirm the suitability of the procedure for the intended purpose. The
response can be demonstrated directly on the product or suitable reference materials, separate
weighings of analyte, or predefined mixtures of the components (e.g., by dilution of a solution of
known content), using the proposed procedure.
Linearity can be evaluated with a plot of signals as a function of analyte concentration or content, and
should demonstrate the analytical procedure capability across a given range to obtain values that are
proportional to the true (known or theoretical) sample values. Test results should be evaluated by an
appropriate statistical method (e.g., by calculation of a regression line by the method of least squares).
Data derived from the regression line may help to provide mathematical estimates of the linearity. A
plot of the data, the correlation coefficient or coefficient of determination, y-intercept and slope of the
regression line should be provided. An analysis of the deviation of the actual data points from the
regression line is helpful for evaluating linearity (e.g., for a linear response, the impact of any non-
random pattern in the residuals plot from the regression analysis should be assessed).
To assess linearity during validation, a minimum of five concentrations appropriately distributed across
the range is recommended.
The measured data can be mathematically transformed if necessary (e.g., through the use of a log
function).
Other approaches to the assessment of linearity should be justified.
3.2.2.2 Non-linear response

Some analytical procedures may show non-linear responses. In these cases, a model or function which
can describe the relationship between the activity/concentration present and the response of the
analytical procedure is necessary. The suitability of the model should be assessed by means of non-
linear regression analysis (e.g., coefficient of determination).
For example, immunoassays or cell-based assays may show an S-shaped response. S-shaped test
curves occur when the range of concentrations is wide enough that responses are constrained by upper
and lower asymptotes. Common models used in this case are four- or five-parameter logistic functions,
though other acceptable models exist.
For these analytical procedures, the evaluation of linearity is separate from consideration of the shape
of the concentration-response curve. Thus, linearity of the concentration-response relationship is not
required. Instead, analytical procedure performance should be evaluated across a given range to
obtain values that are proportional to the true (known or theoretical) sample values.
3.2.2.3 Multivariate calibration

Algorithms used for construction of multivariate calibration models can be linear or non-linear, as long
as the model is appropriate for establishing the relationship between the signal and the quality
attribute of interest. The accuracy of a multivariate procedure is dependent on multiple factors, such as

the distribution of calibration samples across the calibration range and the reference analytical
procedure error.
In multivariate analysis, the measured data are commonly pre-treated through derivatives or
normalisation.
Linearity assessment, apart from comparison of reference and predicted results, should include
information on how the analytical procedure error (residuals) changes across the calibration range.
Graphical plots can be used to assess the residuals of the model prediction across the working range.
3.2.3 Validation of lower range limits
If the quality attribute to be measured requires the range of an analytical procedure to be close to the
lower range limits of the procedure, detection limit (DL) and quantitation limit (QL), can be estimated
using the following approaches.
3.2.3.1 Based on visual evaluation

Visual evaluation can be used for both non-instrumental and instrumental procedures.
The limit is determined by the analysis of samples with known concentrations and by establishing the
minimum level at which the analyte can be reliably resolved and detected or quantitated.
3.2.3.2 Based on signal-to-noise

This approach is relevant for analytical procedures which exhibit baseline noise. Determination of the
signal-to-noise ratio is performed by comparing measured signals from samples with known low
concentrations of analyte with those of blank samples. Alternatively, signals in an appropriate baseline
region can be used instead of blank samples. The DL or QL are the minimum concentrations at which
the analyte can be reliably detected or quantitated, respectively. A signal-to-noise ratio of 3:1 is
generally considered acceptable for estimating the DL. For QL, a ratio of at least 10:1 is considered
acceptable.
The signal-to-noise ratio should be determined within a defined region and, if possible, situated equally
around the place where the peak of interest would be found.
3.2.3.3 Based on the standard deviation of a linear response and a slope

The detection limit (DL) can be expressed as:
3.3𝜎
𝐷𝐿 =
𝑆
while the quantitation limit (QL) can be expressed as:
10𝜎
𝑄𝐿 =
𝑆
where σ = the standard deviation of the response
S = the slope of the calibration curve
The slope S can be estimated from the regression line of the analyte. The estimate of σ can be carried
out in a variety of ways, for example:
Based on the Standard Deviation of the Blank
Measurement of the magnitude of background response is performed by analysing an appropriate

number of blank samples and calculating the standard deviation of the responses.

Based on the Calibration Curve
A specific calibration curve should be evaluated using samples containing an analyte in the range of
the DL and QL. The residual standard deviation of a regression line (i.e., root mean square
error/deviation) or the standard deviation of y-intercepts of the regression lines can be used as the
standard deviation.
3.2.3.4 Based on accuracy and precision at lower range limits

Instead of using estimated values as described in the previous approaches, the QL can be directly
validated by accuracy and precision measurements.
3.2.3.5 Recommended data

The DL and the approach used for its determination should be presented. If the DL is determined
based on visual evaluation or based on signal-to-noise ratio, the presentation of the relevant data is
considered an acceptable justification.
In cases where an estimated value for the DL is obtained by calculation or extrapolation, this estimate
can subsequently be validated by the independent analysis of a suitable number of samples known to
be near or prepared at the DL.
The QL and the approach used for its determination should also be presented.
If the QL was estimated, the limit should be subsequently validated by the analysis of a suitable
number of samples known to be near or at the QL. In cases where the QL is well below (e.g.,
approximately 10 times lower than) the reporting limit, this confirmatory validation can be omitted
with justification.
For impurity tests, the QL for the analytical procedure should be equal to or below the reporting
threshold.
3.3 Accuracy and precision
Accuracy and precision can be evaluated independently, each with a predefined acceptance criterion.
Alternatively, accuracy and precision can be evaluated in combination.
3.3.1 Accuracy
Accuracy should be established across the reportable range of an analytical procedure and is typically
demonstrated through comparison of the measured results with expected values. Accuracy should be
demonstrated under regular test conditions of the analytical procedure (e.g., in the presence of sample
matrix and using described sample preparation steps).
Accuracy is typically verified through one of the studies described below. In certain cases, accuracy can
be inferred once precision, response within the range and specificity have been established.
3.3.1.1 Reference material comparison

The analytical procedure is applied to an analyte of known purity (e.g., a reference material, a well
characterised impurity or a related substance) and the measured versus theoretically expected results
are evaluated.
3.3.1.2 Spiking study

The analytical procedure is applied to a matrix of all components except the analyte where a known
amount of the analyte of interest has been added. In cases where all the expected components are
impossible to reproduce, the analyte can be added to or enriched in the test sample. The results from
measurements on unspiked and spiked/enriched samples are evaluated.

3.3.1.3 Orthogonal procedure comparison
The results of the proposed analytical procedure are compared with those of an orthogonal procedure.
The accuracy of the orthogonal procedure should be reported. Orthogonal procedures can be used with
quantitative impurity measurements to verify primary measurement values in cases where obtaining
samples of all relevant components needed to mimic the matrix for spiking studies is not possible.

Accuracy should be assessed using an appropriate number of determinations and concentration levels
covering the reportable range (e.g., 3 concentrations/3 replicates each of the full analytical procedure).
Accuracy should be reported as the mean percent recovery of a known added amount of analyte in the
sample or as the difference between the mean and the accepted true value, together with an
appropriate 100(1-α) % confidence interval (or justified alternative statistical interval). The observed
interval should be compatible with the corresponding accuracy acceptance criteria, unless otherwise
justified.
For impurity tests, the approach for the determination of individual or total impurities should be
described (e.g., weight/weight or area percent with respect to the major analyte).
For quantitative applications of multivariate analytical procedures, appropriate metrics, e.g., root
mean-squared error of prediction (RMSEP), should be used. If RMSEP is found to be comparable to
acceptable root mean-squared error of calibration (RMSEC) then this indicates that the model is
sufficiently accurate when tested with an independent test set. Qualitative applications such as
classification, misclassification rate or positive prediction rate can be used to characterise accuracy.
3.3.2 Precision
Validation of tests for assay and for quantitative determination of impurity (purity) includes an
investigation of precision.
Precision should be investigated using authentic homogeneous samples or, if unavailable, artificially
prepared samples (e.g., spiked matrix mixtures or samples enriched with relevant amounts of the
analyte in question).
3.3.2.1 Repeatability
Repeatability should be assessed using:
a) a minimum of 9 determinations covering the reportable range for the procedure (e.g., 3
concentrations/3 replicates each)
or
b) a minimum of 6 determinations at 100% of the test concentration.
3.3.2.2 Intermediate precision

The extent to which intermediate precision should be established depends on the circumstances under
which the procedure is intended to be used. The applicant should establish the effects of random
events on the precision of the analytical procedure. Typical variations to be studied include different
days, environmental conditions, analysts and equipment, as relevant. Ideally, the variations tested
should be based on and justified by using analytical procedure understanding from development and
risk assessment (ICH Q14). Studying these effects individually is not necessary. The use of design of
experiments studies is encouraged.

3.3.2.3 Reproducibility
Reproducibility is assessed by means of an inter-laboratory trial. Investigation of reproducibility is
usually not required for regulatory submission but should be considered in cases of standardisation of
an analytical procedure, for instance, for inclusion of procedures in pharmacopoeias and in cases where
analytical procedures are conducted at multiple sites.

The standard deviation, relative standard deviation (coefficient of variation), and an appropriate
100(1-α) % confidence interval (or justified alternative statistical interval) should be reported. The
observed interval should be compatible with the corresponding precision acceptance criteria, unless
otherwise justified.
Additionally, for multivariate analytical procedures, the routine metrics of RMSEP encompass accuracy
and precision.
3.3.3 Combined approaches for accuracy and precision
An alternative to separate evaluation of accuracy and precision is to consider their total impact by
assessing against a combined performance criterion.
Data generated during development may help determine the best approach and refine appropriate
performance criteria to which combined accuracy and precision are compared.
Combined accuracy and precision can be evaluated by use of a prediction interval, a tolerance interval
or a confidence interval. Other approaches may be acceptable if justified.

If a combined performance criterion is chosen, results should be reported as a combined value to
provide appropriate overall knowledge of the suitability of the analytical procedure. If relevant to
justify the suitability of the analytical procedure, the individual results for accuracy and precision
should be provided as supplemental information. The approach used should be described.
3.4 Robustness
The evaluation of the analytical procedure’s suitability within the intended operational environment
should be considered during the development phase and depends on the type of procedure under
study. Robustness testing should show the reliability of an analytical procedure in response to
deliberate variations in analytical procedure parameters, as well as the stability of the sample
preparations and reagents for the duration of the procedure, if appropriate. The robustness evaluation
can be submitted as part of development data for an analytical procedure on a case-by-case basis or
should be made available upon request.
For further details, see ICH Q14.
4. Glossary
Accuracy
The accuracy of an analytical procedure expresses the closeness of agreement between the value
which is accepted either as a conventional true value or as an accepted reference value and the value
or set of values measured. (ICH Q2)

Analytical procedure
The analytical procedure refers to the way of performing the analysis. The analytical procedure should
describe in sufficient detail the steps necessary to perform each analytical test. (ICH Q2)
Analytical procedure parameter
Any analytical factor (including reagent quality) or analytical procedure operational condition that can
be varied continuously (e.g., flow rate) or specified at controllable, unique levels. (ICH Q14)
Analytical procedure validation strategy
An analytical procedure validation strategy describes the selection of analytical procedure performance
characteristics for validation. In the strategy, data gathered during development studies and system
suitability tests (SSTs) can be applied to validation and an appropriate set of validation tests can be
predefined. (ICH Q14)
Calibration model
A model based on analytical measurements of known samples that relates the input data to a value for
the property of interest (i.e., the model output). (ICH Q2)
Control strategy
A planned set of controls, derived from current product and process understanding, that assures
process performance and product quality. The controls can include parameters and attributes related
to drug substance and drug product materials and components, facility and equipment operating
conditions, in-process controls, finished product specifications, and the associated methods and
frequency of monitoring and control. (ICH Q10)
Co-validation
Demonstration that the analytical procedure meets its predefined performance criteria when used at
different laboratories for the same intended purpose. Co-validation can involve all (full revalidation) or
a subset (partial revalidation) of performance characteristics potentially impacted by the change in
laboratories. (ICH Q2)
Detection limit (DL)
The detection limit is the lowest amount of an analyte in a sample which can be detected but not
necessarily quantitated as an exact value. (ICH Q2)
Determination
The reported value(s) from single or replicate measurements of a single sample preparation as per the
validation protocol. (ICH Q2)
Intermediate precision
Intermediate precision expresses intra-laboratory variations. Factors to be considered should include

potential sources of variability, for example, different days, different environmental conditions,
different analysts and different equipment. (ICH Q2)
Performance characteristic
A technology independent description of a characteristic that ensures the quality of the measured
result. Typically, accuracy, precision, specificity/selectivity and range may be considered. Previous ICH
Q2 versions referred to this as Validation characteristic. (ICH Q2)

Performance criterion
An acceptance criterion describing a numerical range, limit or desired state to ensure the quality of the
measured result for a given performance characteristic. (ICH Q14)
Platform analytical procedure
An analytical procedure that is suitable to test quality attributes of different products without
significant change to its operational conditions, system suitability and reporting structure. This type of
analytical procedure can be used to analyse molecules that are sufficiently alike with respect to the
attributes that the platform analytical procedure is intended to measure. (ICH Q2)
Precision
The precision of an analytical procedure expresses the closeness of agreement (degree of scatter)
between a series of measurements obtained from multiple samplings of the same homogeneous
sample under the prescribed conditions. Precision can be considered at three levels: repeatability,
intermediate precision and reproducibility.
The precision of an analytical procedure is usually expressed as the variance, standard deviation or
coefficient of variation of a series of measurements. (ICH Q2)
Quantitation limit (QL)
The quantitation limit is the lowest amount of analyte in a sample which can be quantitatively
determined with suitable precision and accuracy. The quantitation limit is a parameter used for
quantitative assays for low levels of compounds in sample matrices, and, particularly, is used for the
determination of impurities and/or degradation products. (ICH Q2)
Range
The range of an analytical procedure is the interval between the lowest and the highest results in
which the analytical procedure has a suitable level of precision, accuracy and response. (ICH Q2)
Reportable range
The reportable range of an analytical procedure includes all values from the lowest to the highest
reportable result for which there is a suitable level of precision and accuracy. Typically, the
reportable range is given in the same unit as the specification acceptance criterion. (ICH Q2)
Working range
A working range corresponds to the lowest and the highest level of the quality attribute to be
measured (e.g., content or purity) as presented to the analytical instrument and for which the
analytical procedure provides reliable results. (ICH Q2)
Reference material
A suitably characterised material, sufficiently homogeneous and stable with regard to one or more
defined attributes, which has been established to be fit for the intended purpose. Reference materials
may include national/international reference standards, pharmacopoeial reference standards, or in-
house primary/secondary reference materials. (ICH Q2)
Repeatability
Repeatability expresses the precision under the same operating conditions over a short interval of
time. Repeatability is also termed intra-assay precision. (ICH Q2)

Reportable result
The result as generated by the analytical procedure after calculation or processing and applying the
described sample replication. (ICH Q2)
Reproducibility
Reproducibility expresses the precision between laboratories (e.g., inter-laboratory studies, usually
applied to standardisation of methodology). (ICH Q2)
Response
The response of an analytical procedure is its ability (within a given range) to obtain a signal which is
effectively related to the concentration (amount) or activity of analyte in the sample by some known
mathematical function. (ICH Q2)
Revalidation
Demonstration that an analytical procedure is still fit for the intended purpose after a change to the
product, process or the analytical procedure itself. Revalidation can involve all (full revalidation) or a
subset (partial revalidation) of performance characteristics. (ICH Q2)
Robustness
The robustness of an analytical procedure is a measure of its capacity to meet the expected
performance criteria during normal use. Robustness is tested by deliberate variations of analytical
procedure parameters. (ICH Q14)
Specificity/ selectivity
Specificity and selectivity are both terms to describe the extent to which other substances interfere
with the determination of an analyte according to a given analytical procedure. Specificity is typically
used to describe the ultimate state, measuring unequivocally a desired analyte. Selectivity is a relative
term to describe the extent to which particular analytes in mixtures or matrices can be measured
without interferences from other components with similar behaviour. (ICH Q2)
System suitability test (SST)
System suitability tests are developed and used to verify that the measurement system and the
analytical operations associated with the analytical procedure are fit for the intended purpose and
increase the detectability of unacceptable performance. (ICH Q14)
Validation study
An evaluation of prior knowledge, data or deliberate experiments (i.e., validation tests) to determine
the suitability of an analytical procedure for the intended purpose. (ICH Q2)
Validation test
Validation tests are deliberate experiments designed to authenticate the suitability of an analytical
procedure for the intended purpose. (ICH Q2)
Multivariate glossary
Calibration set
A set of data with matched known characteristics and measured analytical results. (ICH Q14)

Independent sample
Independent samples are samples not included in the calibration set of a multivariate model.
Independent samples can come from the same batch from which calibration samples are selected.
(ICH Q2)
Internal testing
Internal testing is a process of checking if unique samples processed by the model yield the correct
predictions (qualitative or quantitative).
Internal testing serves as means to establish the optimal number of latent variables, estimate the
standard error and detect potential outliers. (ICH Q2)
Latent variables
Mathematically derived variables that are directly related to measured variables and are used in further
processing. (ICH Q2)
Model validation
The process of determining the suitability of a model by challenging it with independent test data and
comparing the results against predetermined performance criteria. (ICH Q2)
Multivariate analytical procedure
An analytical procedure where a result is determined through a multivariate calibration model utilising
more than one input variable. (ICH Q2)
Reference analytical procedure
A separate analytical procedure used to obtain the reference values of the calibration and validation
samples for a multivariate analytical procedure. (ICH Q2)
Validation set
A set of data used to give an independent assessment of the performance of the calibration model.
(ICH Q2)
5. References
ICH Q10 Pharmaceutical quality system
ICH Q14 Analytical procedure development
ICH M4Q The common technical document for the registration of pharmaceuticals for human use

ANNEX 1: SELECTION OF VALIDATION TESTS
Figure 2: Examples of relevant validation tests based on the objective of the analytical procedure

Annex 2: Illustrative examples for analytical techniques
The tables presented in this annex are examples of approaches to analytical procedure validation for a
selection of technologies. The technologies and approaches presented have been constructed to
illustrate potential applications of the principles contained within this guideline and are not exhaustive.
The examples are not intended to be mandatory, and alternative approaches (fulfilling the intent of the
guideline) may also be acceptable.
Table 3: Examples for quantitative separation techniques
Technique Separation techniques (e.g., Separation techniques with relative

HPLC, GC, CE) for impurities or area quantitation (e.g., product-
assay related substances such as charge
variants)
Performance Validation study methodology

characteristic
Specificity/ Absence of relevant interference: Absence of relevant interference:

Selectivity
With product, buffer, or appropriate With product, buffer, or appropriate
matrix, and between individual peaks matrix, and between individual peaks of
of interest interest
Spiking with known impurities/ Demonstration of stability-indicating

excipients properties through appropriate forced
degradation samples if necessary
or
By comparison of impurity profiles by

an orthogonal analytical procedure
Demonstration of stability-indicating
properties through appropriate forced
degradation samples, if necessary
Precision Repeatability:
Replicate measurements with 3 times 3 levels across the reportable range or 6

times at 100% level, considering peak(s) of interest
Intermediate precision:
e.g., different days, environmental conditions, analysts, equipment
Accuracy For Assay: Comparison with an orthogonal procedure

and/or suitably characterised material
Comparison with suitably
(e.g., reference material)
characterised material (e.g.,
reference material) or
or Accuracy can be inferred once precision,

linearity and specificity have been
Comparison with an orthogonal
established
procedure

Technique Separation techniques (e.g., Separation techniques with relative
HPLC, GC, CE) for impurities or area quantitation (e.g., product-
assay related substances such as charge
variants)

characteristic
For impurities or related substances: or
Spiking studies with impurities Spiking studies with forced degradation

samples and/or suitably characterised
or
material
Comparison of impurity profiles with
an orthogonal procedure
Reportable Range Validation of calibration model across Validation of calibration model across the
the range: range:
Linearity: Dilution of the analytes of Linearity: Between measured (observed)

interest over the expected procedure relative result versus theoretically
range, at least 5 points expected relative result across
specification range(s), e.g., by spiking or
Validation of lower range limits (for
degrading material
purity only): QL, DL through a
selected methodology (e.g., signal- Validation of lower range limits: QL (and
to-noise determination) DL) through a selected methodology (e.g.,
signal-to-noise determination)
Robustness and Deliberate variation of relevant parameters, e.g.,

other considerations
Sample preparation: extraction volume, extraction time, temperature, dilution
(performed as part
of analytical Separation parameters: column/capillary lot, mobile phase/buffer composition
procedure and pH, column/capillary temperature, flow rate, detection wavelength
development as per
Stability of sample and reference material preparations
ICH Q14)
Relative Response Factors
If the analyte has a different response from the reference material (e.g., a
different specific UV absorbance), relative response factors should be calculated
using the appropriate ratio of responses. This evaluation may be performed
during validation or development, and should use the finalised analytical
procedure conditions and be appropriately documented
If the relative response factor is outside the range 0.8-1.2, then a correction
factor should be applied. If an impurity/degradation product is overestimated, it
may be acceptable not to use a correction factor

Table 4: Example for elemental impurities by ICP-OES or ICP-MS
Technique Elemental Impurities by ICP-OES or ICP-MS

characteristic
Specificity/ Spiking of elements into matrix and demonstration of sufficient non-interference

Selectivity and confirmation of accuracy with the presence of components (e.g., carrier gas,
impurities, matrix)
or
Justification through technology/prior knowledge (e.g., specificity of technology

for certain isotopes)
Replicate measurements with 3 times 3 levels across the reportable range or 6

times at 100% level, considering signals of interest
e.g., different days, environmental conditions, analysts, equipment
Accuracy Spiking studies with impurities
or
Comparison of impurity profiles with an orthogonal procedure
Reportable Range Validation of working range:
Linearity: Dilution of the analytes of interest over the expected working range, at
least 5 points, can be combined with multi-level accuracy experiment
Validation of lower range: QL, DL through a selected methodology
Robustness and Deliberate variation of parameters and stability of test conditions, e.g.,
Sample digestion technique and preparation, nebulizer and sheath flow settings,
(performed as part
plasma settings
of analytical
procedure
development as per
ICH Q14)

Table 5: Example for dissolution with HPLC as product performance test for an immediate release
dosage form
Technique Dissolution with HPLC as product performance test for an immediate

release dosage form
Performance Demonstration of performance of Validation testing methodology

characteristic dissolution step
Typically demonstrated with final
Typically demonstrated with procedure
development data
Specificity/ Discriminatory power: Absence of interference:

Selectivity
Demonstration of the discriminatory Demonstration of non-interference with
power to differentiate between excipients and dissolution media likely
batches manufactured with different to impact the quantitation of the main
critical process parameters and/or analyte
critical material attributes which may
have an impact on the bioavailability
(performed as part of development of
dissolution step)
Precision Repeatability and intermediate Repeatability and intermediate

precision: precision:
Understanding of variability by Demonstration with a homogeneous

performing, e.g., vessel-to-vessel sample from one dissolved tablet, e.g.,
repeatability studies or intermediate several samples drawn from the same
precision studies (operators, vessel, after analyte in sample has been
equipment) fully dissolved
Note: The study provides a combined

assessment of variability of product
quality and product dissolution
performance in addition to the
variability of the quantitative
procedure
Accuracy (Not applicable for dissolution step) Spiking study:
Add known amounts of the reference

material to the dissolution vessel
containing excipient mixture in
dissolution media and calculate recovery
within defined working range

Technique Dissolution with HPLC as product performance test for an immediate
release dosage form
Performance Demonstration of performance of Validation testing methodology

characteristic dissolution step
Typically demonstrated with final
Typically demonstrated with procedure
development data
Reportable Range (Not applicable for dissolution step) Validation of calibration model across
the range
Linearity:
Demonstrate linearity from sample

concentrations (as presented to
quantitative measurement) in the range
of Q - 45% of the lowest strength up to
130% of the highest strength, for one
point specification, and in the range of
QL up to 130% of the highest strength,
for multiple point specification
If lower concentration ranges are

expected to be close to QL:
Validation of lower range limits, see

separation techniques
Robustness and Justification of the selection of the Deliberate variation of parameters of

other dissolution procedure parameters, the quantitative procedure, see
considerations e.g., medium buffer composition, separation technique
(performed as surfactant concentration, use of
part of analytical sinkers, pH, deaeration, volume,
procedure agitation rate, sampling time
development as
per ICH Q14)

Table 6: Example for quantitative 1H-NMR for the assay of a drug substance
Technique Quantitative 1H-NMR (internal standard method) for the assay of a

drug substance

characteristic
Specificity/ Absence of interference:

Selectivity
Select a signal which is representative for the analyte and does not show
interference with potential baseline artefacts, residual water or solvent
signals, related structure impurities or other impurities, internal standards,
non-target major component or potential isomers/forms
Replicate measurements of at least 6 independent preparations at 100% level
Intermediate Precision:
Not necessary to be conducted on target analyte (justified by technology

principle, as typically verified through instrument calibration with a standard
sample)
Accuracy Reference material comparison:
Confirm with sample of known purity
Reportable Range Validation tests are typically not necessary because the integral areas are
usually directly proportional to the amount (mole) of reference material and
analyte (technology inherent justification)
Robustness and Deliberate variation of parameters, e.g.,

other
Temperature, concentration, field (shim), tuning and matching of the NMR
considerations
probe, solution stability
(performed as part
of analytical
procedure
development as per
ICH Q14)

Table 7: Example for biological assays
Technique Binding assay (e.g., ELISA, SPR) or cell-based assay for determination
of potency relative to a reference

characteristic

Selectivity
Dose-response curve fulfils the response criteria demonstrating the similarity
of the analyte and reference material, as well as non-interfering signal from
the matrix (for binding assay), or no dose-response from the cell line alone
(for cell-based assay)
Demonstration of stability-indicating properties through appropriate forced

degradation samples if necessary
Repeated sample analysis on a single day or within a short interval of time

covering the reportable range of the analytical procedure (at least 3 replicates
over at least 5 levels)
Intermediate Precision: Different analysts, multiple independent preparations

over multiple days at multiple potency levels through the analytical
procedure's reportable range, inclusive of normal laboratory variation
Accuracy Reference material comparison:
Assess recovery versus theoretical activity for multiple (at least 3)

independent preparations at multiple (at least 5) levels through the analytical
procedure's reportable range
Reportable Range Validation of range, including lower and higher range limits:
The lowest to highest relative potency levels that meet accuracy, precision,
and response criteria, determined over at least 5 potency levels.

other
Plate type, buffer components, incubation times, incubation conditions,
considerations
instruments, reaction times, reagent lots including controls
(performed as part
of analytical For binding assay procedures: coating proteins, capture/detection antibody
procedure
For cell-based assay procedures: cell density, effector/target cell ratio, cell
development as per
generation number
ICH Q14)

Table 8: Example for quantitative PCR
Technique Quantitative PCR (quantitative analysis of impurities in drug

substances or products)

characteristic
Specificity/ Selectivity Orthogonal Procedure Comparison:
Test reaction specificity by gel electrophoresis, melting profile or DNA

sequencing
Absence of interference:
Positive template, no-reverse transcription control for RT-qPCR and no

template control. Test primer and probe target specificity against gene bank
with sequence similarity search program (e.g., nucleotide BLAST). Evaluate
the slope of standard curve for efficiency
Independent preparations of 5 positive control levels evenly distributed

along the standard curve and assayed in triplicate within a single assay
assessment. The results can be compared using coefficient of variation (CV)
At least three replicates per run at each positive control level in at least 6
runs over two or more days
Accuracy Spiking Study:
Test (e.g., n=6) replicates at 3 to 5 template spike levels from the standard
curve concentrations
Efficiency/consistency of RNA/DNA extraction method should be accounted

for
Reportable Range Linearity:
Working range should cover at least 5 to 6 log to the base 10 concentration

values. Correlation coefficients or standard deviations should be calculated
through the entire dynamic range
Validation of lower working range limits based on the calibration curve:
DL defined by template spiking in samples or from standard curves. DL is

lowest point meeting the response curve parameters
QL demonstrated through showing sufficient recovery and acceptable CVs

from the accuracy experiment
Robustness and other Deliberate variation of parameters, e.g.,

considerations (performed
Equipment, master mix composition (concentrations of salts, dNTPs,
as part of analytical
adjuvants), master mix lots, reaction volume, probe and primer
procedure development as
concentrations, thermal cycling parameters
per ICH Q14)

Table 9: Example for particle size measurement
Technique Particle size measurement
(dynamic light scattering; laser diffraction measurement) as a

property test

characteristic

Selectivity
Evaluate blank and sample to determine the appropriateness of the
equipment settings and sample preparation
Test at least 6 replicates using established analytical procedure parameters

at target range
Analysis performed on different days, environmental conditions, analysts,

equipment setup
Accuracy Technology inherent justification:
Confirmed by an appropriate instrument qualification
or
Orthogonal procedure comparison:
Qualitative comparison using a different technique, like optical microscopy,

to confirm results
Reportable Range Technology specific justification, e.g., particle size range covered

Evaluation of expected size ranges for the intended use of the analytical
(performed as part
procedure
of analytical
procedure Dispersion stability for liquid dispersions (stability over potential analysis
development as per time, stir rate, dispersion energy equilibration or stir time before
ICH Q14) measurement)
Dispersion stability for dry dispersions (sample amount, measurement time,

air pressure and feed rate)
Obscuration range (establish optimum percentage of laser obscuration)

Ultrasound time/percentage for sample, if applicable

Table 10: Example for NIR analytical procedure
Technique NIR analytical procedure for core tablet assay

characteristic

Selectivity
Comparison of drug substance spectrum and the loading plots of the model
Rejection of outliers (e.g., excipient, analogues) not covered by the

multivariate procedure
Repeated analysis with removal of sample from the holder between

measurements
Accuracy Comparison with an orthogonal procedure:
Demonstration across the range through comparison of the predicted and

reference values using an appropriate number of determinations and
concentration levels (e.g., 5 concentrations, 3 replicates)
Accuracy is typically reported as the standard error of prediction (SEP or

RMSEP)
Reportable Range Response:
Demonstration of the relationship between predicted and reference values
Error (accuracy) across the range:
Information on how the analytical procedure error (accuracy) changes

across the calibration range, e.g., by plotting the residuals of the model
prediction versus the actual data

Chemical and physical factors that can impact NIR spectrum and model
(performed as part
prediction should be represented in data sets. Examples include various
of analytical
sources of drug substance and excipients, water content, tablet hardness,
procedure
and orientation in the holder
development as per
ICH Q14) Note: NIR measurements are sensitive to changes in tablet composition and
properties outside variation present in the calibration set

Table 11: Example for quantitative LC/MS
Technique Quantitative LC/MS analysis of trace impurities in product

characteristic
Specificity/ Technology inherent justification:

Selectivity
Inferred through use of specific and selective MS detection (e.g., MRM
transition with specified quantitative to qualitative ion ratio, accurate m/z
value) in combination with retention time, consider potential for isotopes
or
Absence of interference from other components in sample matrix
or
Comparison of impurity profiles determined by an orthogonal analytical

procedure
Precision Repeatability
Measurement of a minimum of 3 replicates at each of at least 3 spiking levels or

a minimum of 6 replicates at 100%
Intermediate precision
Comparison of measurements of the same samples performed in the same

laboratory but under varying conditions (e.g., different LC/MS systems,
different analysts, different days)
Accuracy Spiking study:
Acceptable recovery of spiked impurity standards in sample matrix at multiple

spiking levels
or
Comparison of the results to the ‘true’ values obtained from an orthogonal

procedure
Reportable Range Validation of calibration model across the range:
Linearity: Experimental demonstration of the linear relationship between

analyte concentrations and peak responses (or the ratio of peak response if an
internal standard was used) with reference materials at 5 or more concentration
levels
Validation of lower range limits:
DL: Use the coefficient of variation (CV) of responses at the spiking level (with
6 or more repeated injections) as a measure of signal-to-noise. The CV
obtained must be less than or equal to a pre-defined acceptable value
QL: The lowest spiking level with acceptable accuracy and precision
The range extends from and is inclusive of the QL to the highest spiking level
with acceptable accuracy, precision and response

Technique Quantitative LC/MS analysis of trace impurities in product

characteristic
Robustness and other Deliberate variation of parameters, e.g.,

considerations
LC flow rate, LC injection volume, MS drying/desolvation temperature, MS gas
(performed as part of
flow, mass accuracy, MS collision energy, stability of test conditions
analytical procedure
development as per
ICH Q14)

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14 December 2023

ICH Q2(R2) Guideline on validation of analytical

Transmission to CHMP 8 March 2022

Adoption by CHMP 24 March 2022

Release for public consultation 31 March 2022

Deadline for comments 31 July 2022

Final adoption by CHMP 14 December 2023

Date for coming into effect 14 June 2024

Official address Domenico Scarlattilaan 6 ● 1083 HS Amsterdam ● The Netherlands

Code History Date

Q2 Approval by the Steering Committee under Step 2 and 26 October 1993

Q2(R2) Complete revision of guideline to include more recent 24 March 2022

Endorsement by the Members of the ICH Assembly under

Q2(R2) Error Correction to Table 5: Dissolution with HPLC as 30 November 2023

EMA/CHMP/ICH/82072/2006 Page 2/33

EMA/CHMP/ICH/82072/2006 Page 3/33

2. General considerations for analytical procedure

EMA/CHMP/ICH/82072/2006 Page 4/33

2.1 Analytical procedure validation study

EMA/CHMP/ICH/82072/2006 Page 5/33

Measured Quality IDENTITY IMPURITY (PURITY) ASSAY

Quantitative Limit Test

Lower Range Limit - QL† DL -

Intermediate - + (5) - + (5)

- signifies that this test is not normally conducted

+ signifies that this test is normally conducted

† in some complex cases DL may also be evaluated

QL, DL: quantitation limit, detection limit

EMA/CHMP/ICH/82072/2006 Page 6/33

Figure 1: Validation study design and evaluation

2.2 Validation during the lifecycle of an analytical procedure

EMA/CHMP/ICH/82072/2006 Page 7/33

2.3 Reportable range

2.4 Demonstration of stability-indicating properties

2.5 Considerations for multivariate analytical procedures

Typically, development and validation are performed in two phases.

EMA/CHMP/ICH/82072/2006 Page 8/33

Potency Lowest specification acceptance Highest specification acceptance

Content uniformity 70% of declared content 130% of declared content

Lower limit of reportable range 130% of declared content of the

Lower limit of reportable range

Impurity (1) Reporting threshold 120% of specification acceptance

Purity (as area %) 80% of lower specification Upper specification acceptance

EMA/CHMP/ICH/82072/2006 Page 9/33

3.1 Specificity/ selectivity

3.1.1 General considerations

The specificity or selectivity of an analytical procedure can be demonstrated through absence of

3.1.1.1 Absence of interference

3.1.1.2 Orthogonal procedure comparison

3.1.1.3 Technology inherent justification

3.1.2 Recommended data

EMA/CHMP/ICH/82072/2006 Page 10/33

Representative data (e.g., chromatograms, electropherograms, spectra, biological response) should be

Impurities or related substances are available or can be intentionally created:

Impurities or related substances are not available:

3.2.1 General considerations

EMA/CHMP/ICH/82072/2006 Page 11/33

3.2.2.1 Linear response

Other approaches to the assessment of linearity should be justified.