ICH Q2R1 guideline of analytical method development and validation

1. Q2R1
BY- ASST. PROF. SANJAYKUMAR UCHIBAGLE,
GHRU, SAIKHEDA.

2. VALIDATION OF ANALYTICAL
PROCEDURES: TEXT AND
METHODOLOGY
Q2(R1)

3. INTERNATIONAL CONFERENCE ON HARMONISATION
OF TECHNICAL REQUIREMENTS FOR REGISTRATION
OF PHARMACEUTICALS FOR HUMAN USE

4. LIST OF ICH GUIDELINE
Q1A (R2) – Stability Testing of New Drug Substances and Products
Q2 (R1) – Validation of Analytical Procedures : Text and Methodology
Q3A (R2) – Impurities in New Drug Substances
Q4 – Pharmacopoeias
Q5B – Quality of Biotechnology Products
Q6A – Specifications : Test Procedure and Acceptance Criteria for New Drug
Substances and New Drug Products: Chemical Substances
Q7 – Good Manufacturing Guide for Active Pharmaceutical Ingredients
Q8(R2) – Pharmaceutical Development
Q9 – Quality Risk Management
Q10 – Pharmaceutical Quality System
Q11 – Development and Manufacture of Drug Substances (Chemical
Entities Biotechnological/Biological Entities)
Q12 – Life Cycle Management
Q13 -EWG Continuous Manufacturing of Drug Substances and Drug Products
Q2(R2)/Q14 – EWG Analytical Procedure Development and Revision of Q2 (R1)
Analytical Validation

5. ICH Q1 UIDELINE
 Q1A (R2) – Stability Testing of New Drug Substances and Products.
 Q1 B – Stability Testing : Photo Stability Testing of New Drug
Substances and Products.
 Q1C – Stability Testing for New Dosage Forms.
 Q1D – Bracketing and Matrixing Designs for Stability Testing of New
Drug Substances and Products.
 Q1E – Evaluation of Stability Data.
 Q1F – Stability Data Package for Registration Application in Climatic
Zones III and IV.

6.  Q2 (R1) – Validation of Analytical Procedures : Text and
Methodology
 Q3A (R2) – Impurities in New Drug Substances
 Q3B (R2) – Impurities in New Drug Products
 Q3C (R5) – Impurities : Guideline for Residual Solvents
 Q3D – Impurities : Guideline for Elemental Impurities

7.  Q4A – Pharmacopoeial Harmonisation
 Q4B – Evaluation and Recommendation of Pharmacopoeial
Text for use in the ICH Regions
 Q4B Annex 1(R1) – Residue on Ignition /Sulphated Ash
General Chapter
 Q4B Annex 2(R1) – Test for Extractable Volume of Parenteral
Preparation General Chapter

8.  Q5A(R1) – Viral Safety Evaluation of Biotechnology Products
Derived from Cell Lines of Human or Animal Origin
 Q5B – Quality of Biotechnology Products
 Q5C – Quality of Biotechnology Products :Quality of
Biotechnological
 Q5D – Derivation and Characterisation of Cell Substrates used
for Production of Biotechnological/Biological Products
 Q5E – Comparability of Biotechnological/Biological Products
Subject to Changes in their Manufacturing Process

9.  Q6A – Specifications : Test Procedure and Acceptance
Criteria for New Drug Substances and New Drug Products:
Chemical Substances.
 Q6B – Specifications : Test Procedure and Acceptance
Criteria for Biotechnological/Biological.
 Q7 – Good Manufacturing Guide for Active Pharmaceutical
Ingredients
 Q8(R2) – Pharmaceutical Development
 Q9 – Quality Risk Management

10.  Q10 – Pharmaceutical Quality System
 Q11 – Development and Manufacture of Drug Substances
(Chemical Entities Biotechnological/Biological Entities)
 Q12 – Life Cycle Management
 Q13 -EWG Continuous Manufacturing of Drug Substances
and Drug Products
 Q2(R2)/Q14 – EWG Analytical Procedure Development and
Revision of Q2 (R1) Analytical Validation

11. STEPES INVOLVED IN THE ICH Q2R1 GUIDELINE
1. SPECIFICITY
2. ACCURACY
3. PRECISION
4. DETECTION LIMIT
5. QUANTITATION LIMIT
6. LINEARITY
7. RANGE
8. ROBUSTNESS
9. SYSTEM SUITABILITY TESTING

12. 1. SPECIFICITY
 Specificity is the ability of a method to detect and differentiate the analyte from other substances,
including its related substances.
 e.g., substances that are structurally similar to the analyte, metabolites, isomers, impurities, degradation
products formed during sample preparation.
 Identification -tests should be able to discriminate between compounds of closely related structures
which are likely to be present.
 Assay and Impurity Test(s)-For critical separations, specificity can be demonstrated by the resolution of
the two components which elute closest to each other.
 Impurities are available-This can be done by spiking pure substances with appropriate levels of
impurities and/or excipients and demonstrating that the assay result is unaffected by the presence of
these materials.
 Impurities are not available- by comparing the test results of samples containing impurities or
degradation products to a second well-characterized procedure e.g.: Pharmacopoeial method or other
validated analytical procedure.

13. 2. ACCURACY (Trueness)
The accuracy of an analytical procedure expresses the closeness of agreement
between the value which is accepted either as a conventional true value or an
accepted reference value and the value found.
DRUG SUBSTANCES & DRUG PRODUCTS.
 Application of an analytical procedure to an analyte of known purity (e.g.
reference material).
 Comparison of the results of the proposed analytical procedure with those of a
second well-characterized procedure, the accuracy of which is stated and/or
defined (independent procedure).
Impurities (Quantitation)
In cases where it is impossible to obtain samples of certain impurities and/or
degradation products, it is considered acceptable to compare results obtained by
an independent procedure.

14. 3. PRECISION
3.1. Repeatability / Intra-assay Precision.
3.2. Intermediate precision
3.3. Reproducibility
3.1. Repeatability / Intra-assay Precision.
Repeatability expresses the precision under the same operating conditions over a short
interval of time.
3.2. Intermediate precision
Intermediate precision expresses within-laboratories variations: different days,
different analysts, different equipment, etc.
3.3. Reproducibility
Reproducibility expresses the precision between laboratories (collaborative studies,
usually applied to standardization of methodology).

15. 3.1. Repeatability / Intra-assay Precision.
Repeatability should be assessed using:
a) a minimum of 9 determinations covering the specified range for the procedure (e.g., 3
concentrations/3 replicates each); or
b) a minimum of 6 determinations at 100% of the test concentration.
3.2. Intermediate precision
Typical variations to be studied include within lab.
 Different days,
 Different analysts,
 Different equipment, etc.
It is not considered necessary to study these effects individually.

16. 3.3. Reproducibility
 Reproducibility is assessed by means of an inter-laboratory trial.
 Reproducibility should be considered in case of the standardization of an
analytical procedure.

17. 4. DETECTION LIMIT
The detection limit of an individual analytical procedure is the lowest amount of analyte in a
sample which can be detected but not necessarily quantitated as an exact value.
4.1. Based on Visual Evaluation
The detection limit is determined by the analysis of samples with known concentrations of
analyte and by establishing the minimum level at which the analyte can be reliably detected.
4.2. Based on Signal-to-Noise
This approach can only be applied to analytical procedures which exhibit baseline noise.
 signal-to-noise ratio is performed by comparing measured signals from samples with known
low concentrations of analyte
 with those of blank samples and establishing the minimum concentration at which the
analyte can be reliably detected.
A signal-to-noise ratio generally considered 3 or 2:1

18. 4.3 Based on the Standard Deviation of the Response and the Slope
The detection limit (DL) may be expressed as:
DL = 3.3 σ / S
where σ = the standard deviation of the response (variety of ways)
S = the slope of the calibration curve of analyte.
4.3.1. Based on the Standard Deviation of the Blank
Measurement of the magnitude of analytical background response is performed by
analysing an appropriate number of blank samples and calculating the standard deviation
of these responses.
4.3.2. Based on the Calibration Curve
The residual standard deviation of a regression line or the standard deviation of y-
intercepts of regression lines may be used as the standard deviation.

19. 5.QUANTITATION LIMIT
The quantitation limit of an individual analytical procedure is the lowest amount of analyte in
a sample which can be quantitatively determined with suitable precision and accuracy.
5.1. Based on Visual Evaluation
The analysis of samples with known concentrations of analyte and by establishing the
minimum level at which the analyte can be quantified with acceptable accuracy and
precision.
5.2. Based on Signal-to-Noise Approach
Applied to analytical procedures that 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 and by establishing the minimum concentration at which the analyte can be
reliably quantified. A typical signal-to-noise ratio is 10:1.

20. 5.3. Based on the Standard Deviation of the Response and the
Slope
The quantitation limit (QL) may be expressed as:
QL = 10 σ /S
where σ = the standard deviation of the response
S = the slope of the calibration curve
σ = the standard deviation of the response can be calculated by using
5.3.1. Based on Standard Deviation of the Blank
5.3.2.Based on the Calibration Curve

21. 6. ROBUSTNESS
The robustness of an analytical procedure is a measure of its capacity to
remain unaffected by small, but deliberate variations in method parameters
and provides an indication of its reliability during normal usage.
Examples of typical variations are:
- stability of analytical solutions;
- extraction time.
In the case of liquid chromatography, examples of typical
variations are:
- influence of variations of pH in a mobile phase;
- influence of variations in mobile phase composition;
- different columns (different lots and/or suppliers);
- temperature;
- flow rate.

22. 7. RANGE
Analytical procedure is the interval between the upper and lower concentration of
analyte in the sample for which it has been demonstrated that the analytical procedure
has a suitable level of precision, accuracy and linearity.
• Assay of a drug substance or finished (drug) product: 80 to 120 %
• for content uniformity, covering a minimum of 70 to 130
• for dissolution testing: +/-20 % over the specified range;
• for the determination of an impurity: from the reporting level of an impurity1 to 120%
of the specification;
• if assay and purity are performed together as one test and only a 100% standard is
used, linearity should cover the range from the reporting level of the impurities1 to
120% of the assay specification.

23. 8. LINEARITY
The linearity of an analytical procedure is its ability (within a given range) to
obtain test results which are directly proportional to the concentration
(amount) of analyte in the sample.
 Linearity should be evaluated by visual inspection of a plot of signals as a function of analyte
concentration or content.
 If there is a linear relationship, test results should be evaluated by appropriate statistical
methods, for example, by calculation of a regression line by the method of least squares.
 The correlation coefficient, y-intercept, slope of the regression line and residual sum of squares
should be submitted. A plot of the data should be included.
 In addition, an analysis of the deviation of the actual data points from the regression line may
also be helpful for evaluating linearity.

24. 9. SYSTEM SUITABILITY TESTING
 System suitability testing is an integral part of many analytical
procedures.
 the equipment, electronics, analytical operations and samples to be
analysed constitute an integral system that can be evaluated as
such.
 System suitability test parameters to be established for a particular
procedure depend on the type of procedure being validated.

25. ANY QUE?

26. Thank You!