Pharma 462 Validation

PHARM 462

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European and International regulatory bodies and their guidelines on different aspects of QA

Body Full name Guidance on

Eurachem Focus for Analytical Chemistry in Europe Method validation

CITAC Cooperation of International Traceability in Analytical Chemistry

Proficiency testingQuality Assurance

EA European Cooperation for Accreditation Accreditation

CEN European Committee for Normalization Standardization

IUPAC International Union of Pure & Applied Chem. Method validation

ISO International Standardization Organisation Standardisation

AOAC

ILAC

Association of Official Analytical Chemists

International Laboratory Accreditation Cooperat.

Internal qual. ControlProficiency testingAccreditation

FDA US Food and Drug Administration Method validation

USP United States Pharmacopoeia Method validation

ICH International Conference on Harmonization Method validation

2 2009

Method Validation

Validation of analytical procedures is the process of determining the

suitability of a given methodology for providing useful

analytical data.

J. Guerra, Pharm. Tech. March 1986

Validation is the formal and systematic proof that a method compiles

with the requirements for testing a product when

observing a defined procedures.

G. Maldener, Chromatographia, July 1989

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Method validation is the process of demonstrating that analytical

procedures are suitable for their intended use and that they support

the identity, strength, quality, purity and potency of the

drug substances and drug products

Method validation is primarily concerned with:

identification of the sources of potential errors

quantification of the potential errors in the method

An method validation describes in mathematical and quantifiable

terms the performance characteristics of an assay

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Examples of Methods That Require

Validation Documentation

Chromatographic Methods - HPLC, GC, TLC, GC/MS, etc.

Pharmaceutical Analysis - In support of CMC.

Bioanalytical Analysis - In support of PK/PD/Clinical Studies.

Spectrophotometric Methods – UV/VIS, IR, NIR, AA, NMR,

XRD,MS

Capillary Electrophoresis Methods - Zone, Isoelectric Focusing

Particle Size Analysis Methods - Laser, Microscopic, Sieving, SEC,

etc.

Automated Analytical Methods - Robots, Automated Analysis.

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Considerations Prior to Method Validation

Suitability of Instrument Status of Qualification and Calibration

Suitability of Materials Status of Reference Standards, Reagents, Placebo Lots

Suitability of Analyst Status of Training and Qualification Records

Suitability of Documentation Written analytical procedure and proper approved protocol

with pre-established acceptance criteria

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Validation Step Define the application, purpose and scope of the method.

Analytes? Concentration? Sample matrices?

Develop a analytical method.

Develop a validation protocol.

Qualification of instrument.

Qualify/train operator

Qualification of material.

Perform pre-validation experiments.

Adjust method parameters and/or acceptance criteria if necessary.

Perform full validation experiments.

Develop SOP for executing the method in routine analysis.

Document validation experiments and results in the validation report.

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Purpose of Method Validation

Identification of Sources and Quantitation of Potential errors

Determination if Method is Acceptable for Intended Use

Establish Proof that a Method Can be Used for Decision Making

Satisfy FDA Requirements

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What is not Analytical Method

Validation?

Calibration

The Process of Performing Tests on Individual System

Components to Ensure Proper function

For example) HPLC Detector calibration

Wavelength Accuracy/ Linear Range/ Noise Level/ Drift

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System Suitability

Test to verify the proper functioning of the operating

system, i.e., the electronics, the equipment, the specimens

and the analytical operations.

Minimum Resolution of 3.0 between the analyte peak and

internal standard peaks

Relative Standard Deviation of replicate standard injections

of not more than 2.0%

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System Suitability

Sample

Validation

MethodAnalyst

Calibration

Pump

Detector

Injector

Data System

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Method Life Cycle

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Validation

Development Optimization

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Verification vs. Validation

Compendial vs. Non-compendial Methods

Compendial methods-Verification

Non-compendial methods-Validation requirement

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Compendial Analytical Procedures

The Analytical procedures in the USP 25/NF 20 are legally recognized under

section 501(b) of the Federal Food, Drug and Cosmetic Act as the regulatory

analytical procedures for the compendial items. The suitability of these

procedures must be verified under actual conditions of use. When using USP

25/NF 20 analytical procedures, the guidance recommends that information be

provided for the following

characteristics:

Specificity of the procedure

Stability of the sample solution

Intermediate precision 142009

Published Validation Guidelines

1978 Current Good Manufacturing Practices (cGMPs)

1987 FDA Validation Guideline

1989 Supplement 9 to USP XXI

1994 CDER Reviewer Guidance:

Validation of Chromatographic Method

1995 ICH Validation Definitions:

Q2A, Text on Validation of Analytical procedures

1997 ICH Validation Methodology:

Q2B, Validation of Analytical Procedures: Methodology

1999 Supplement 10 to USP 23 <1225>: Validation of Compendial Methods

1999 CDER “Bioanalytical Method Validation for Human Studies”

2000 CDER Draft “Analytical Procedures and Method Validation”152009

Regulatory and Compliance Requirements Review

Validation of an analytical method is the

process by which it is established, by

laboratory studies, that the performance

characteristics of the method meet the

requirements for the intended analytical applications

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USP 23 General Information <1225>

2009

The accuracy, sensitivity, specificity, and

reproducibility of test methods employed by the firm

shall be established and documented. Such validation

and documentation may be accomplished in

accordance with 211.194(a)(2).

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21 CFR PART 211 - CURRENT GOOD

MANUFACTURING PRACTICE FOR FINISHED

PHARMACEUTICALS

Subpart I-Laboratory Controls

211.165 Testing and release for distribution (e)

2009

The objective of validation of an analytical

procedure is to demonstrate that it is suitable

for its intended purpose

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ICH Guideline for Industry Q2A, Text on Validation of Analytical ProceduresMarch 1995

2009

In practice, it is usually possible to design the experimental work such that the appropriate validation characteristics can be considered simultaneously to provide a sound, overall knowledge of the capabilities of the analytical procedure, for instance: Specificity, Linearity, Range, Accuracy, and

Precision.

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ICH Guideline for Industry Q2B, Validation of Analytical Procedures: Methodology

2009

Today’s Validation Requirements

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ICH/USP

GMPs(legal) FDA

2009

ICH/USP Validation Requirements & Parameters

Specificity

Linearity

Range

Accuracy

Precision Repeatability Intermediate Precision Reproducibility

Limit of Detection

Limit of Quantitation21

ICH

SpecificitySpecificity Linearity and RangeLinearity and Range AccuracyAccuracy Precision Precision Limit of DetectionLimit of Detection Limit of QuantitationLimit of Quantitation RuggednessRuggedness RobustnessRobustness

USP

2009

USP Data Elements Required For Assay Validation

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Analytical PerformanceParameter

Assay Category

1

Assay Category 2Assay

Category 3

Quantitative

Limit Tests

Accuracy Yes Yes * *

Precision Yes Yes No Yes

Specificity Yes Yes Yes *

LOD No No Yes *

LOQ No Yes No *

Linearity Yes Yes No *

Range Yes Yes * *

Ruggedness Yes Yes Yes Yes

* May be required, depending on the nature of the specific test.2009

USP Categories

Category 1: Quantitation of major components or

active ingredients

Category 2: Determination of impurities or

degradation products

Category 3: Determination of performance

characteristics

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ICH Validation Characteristics vs. Type of Analytical Procedure

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Type of Analytical Procedure

Identification

Impurity testing

AssayQuantitative

Limit Tests

Accuracy No Yes No Yes

Precision

Repeatability

No Yes No Yes

Interm. Prec.

No Yes No Yes

Specificity Yes Yes Yes Yes

LOD No No Yes No

LOQ No Yes No No

Linearity No Yes No Yes

Range No Yes No Yes

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Specificity/Selectivity Ability of an analytical method to measure the analyte free from

interference due to other components.

Selectivity describes the ability of an analytical method to differentiate

various substances in a sample

Original term used in USP

Also Preferred by IUPAC and AOAC

Also used to characterize chromatographic columns

Degree of Bias (Used in USP)

The difference in assay results between the two groups

- the sample containing added impurities, degradation products, related chemical

compounds, placebo ingredients

- the sample without added substances 252009

Specificity: Impurities Assay

Chromatographic Methods Demonstrate Resolution

Impurities/Degradants Available Spike with impurities/degradants

Show resolution and a lack of interference

Impurities/Degradants Not Available Stress Samples

For assay, Stressed and Unstressed Samples should be

compared.

For impurity test, impurity profiles should be compared.

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Forced Degradation Studies

Temperature (50-60 )℃

Humidity (70-80%)

Acid Hydrolysis (0.1 N HCl)

Base Hydrolysis (0.1 N NaOH)

Oxidation (3-30%)

Light (UV/Vis/Fl)

Intent is to create 10 to 30 % Degradation

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Linearity

Ability of an assay to

elicit a direct and

proportional response

to changes in analyte

concentration.

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Linearity Should be Evaluated

By Visual Inspection of plot of signals vs. analyte

concentration

By Appropriate statistical methods

Linear Regression (y = mx + b)

Correlation Coefficient, y-intercept (b), slope (m)

Acceptance criteria: Linear regression r2 > 0.95

Requires a minimum of 5 concentration levels

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Range Acceptable range having linearity, accuracy, precision.

For Drug Substance & Drug product Assay

80 to 120% of test Concentration

For Content Uniformity Assay

70 to 130% of test Concentration

For Dissolution Test Method

+/- 20% over entire Specification Range

For Impurity Assays

From Reporting Level to 120% of Impurity Specification for Impurity

Assays

From Reporting Level to 120% of Assay Specification for Impurity/Assay

Methods302009

Accuracy

Closeness of the test

results obtained by the

method to the true value.

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Accuracy Should be established across specified range of

analytical procedure.

Should be assessed using a minimum of 3 concentration

levels, each in triplicate (total of 9 determinations)

Should be reported as:

Percent recovery of known amount added or

The difference between the mean assay result and the accepted

value

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Accuracy Data Set (1 of 3)

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Amount Added (mg)

AmountFound (mg)

Percent Recovery

0.0 0.0 ---

50.2 50.4 100.5

79.6 80.1 100.6

99.9 100.7 100.8

120.2 119.8 99.7

150.4 149.7 99.5

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Precision

The closeness of agreement (degree of

scatter) between a series of

measurements obtained from

multiple samplings of the same

homogeneous sample.

Should be investigated using

homogeneous, authentic samples.

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Precision… Considered at 3 Levels

Repeatability

Intermediate Precision

Reproducibility

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Repeatability

Express the precision

under the same

operating conditions

over a short interval of

time.

Also referred to as

Intra-assay precision

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Should be

assessed using

minimum of 9

determinations

(3

concentrations/ 3

replicates) or or

Minimum of 6

determinations at

the 100% level.2009

Intermediate Precision

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Express within-

laboratory variations.

Expressed in terms of

standard deviation,

relative standard

deviation (coefficient

of variation) and

confidence interval.

Depends on the

circumstances under

which the procedure

is intended to be

used.

Studies should

include varying days,

analysts, equipment,

etc.2009

Repeatability & Intermediate Precision

Day 1 Day 2100.6 99.5

100.8 99.9

100.1 98.9

100.3 99.2

100.5 99.7

100.4 99.6

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GrandMean = 100.0RSD = 0.59%

Mean = 100.5RSD = 0.24%

Mean = 99.5RSD = 0.36%

2009

Reproducibility

Definition: Ability reproduce data

within the predefined precision

Determination: SD, RSD and

confidence interval

Repeatability test at two different

labs.

Note: Data not required for BLA/NDA

Lab 1 Lab 2 Lab 3

Day 1

Day 2

Day 1

Day 2

Day 1

Day 2

Man 1

Man 2

Man 1

Man 2

Man 1

Man 2

3 Prep

3 Prep

3 Prep

3 Prep

3 Prep

3 Prep

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Detection Limit (LOD)/ Quantitation Limit (LOQ)

LOD

Lowest amount of analyte in a

sample that can be detected

but not necessarily

quantitated.

Estimated by Signal to Noise

Ratio of 3:1.

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LOQ

Lowest amount of

analyte in a sample

that can be quantified

with suitable accuracy

and precision.

Estimated by Signal to

Noise Ratio of 10:1. 2009

1. Based in Visual Evaluations

- Used for non-instrumental methods

2. Based on Signal-to Noise-Ratio

- 3:1 for Detection Limit

- 10:1 for Quantitation Limit

3. Based on Standard Deviation of the Response and

the Slope

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LOD and LOQ Estimated by

2009

S = slope of calibration curve

s = standard deviation of blank readings or

standard deviation of regression line

Validated by assaying samples at DL or QL 42

DL DL ==

3.3s3.3sQL =QL =

10s10s

SS SS

LOD and LOQ Estimated by

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Ybl

LOD LOQ

Statistical estimate of LOD & LOQ

LOD = 3.3 Sbl / b LOQ = 10 Sbl / b

Y = b X + a

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Definition: Capacity to remain unaffected by small but deliberate

variations in method parameters

Determination: Comparison results under differing conditions

with precision under normal conditions

Examples of typical variations in LC

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

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Robustnes

s

2009

Ruggedness

Degree of reproducibility of test results

under a variety of conditions

Different Laboratories

Different Analysts

Different Instruments

Different Reagents

Different Days

Etc.

Expressed as %RSD452009

ICH/USP System Suitability

ICH

Definition: evaluation of equipment, electronic,

analytical operations and samples as a whole

Determination: repeatability, tailing factor (T), capacity

factor (k’), resolution (R), and theoretical Plates (N)

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USP 23 <621> System Suitability Requirements

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Parameters

Recommendations

K’ In general k’ ≥ 2.0

R

R > 2, between the peak of interest and the closest potential interferent (degradant, internal STD, impurity, excipient, etc…..)

T T ≤ 2

N In general N > 2000

Repeatability

RSD ≤ 2.0% (n ≥ 5)

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Re-validation When

Method parameters have been changed The scope of the method has been changed Synthetic methods have been changed Impurity profile has been changed

What Preferably everything. Exceptions should be

scientifically justified

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How do we Know the expectations of the FDA?

FDA Form 483

FDA Warning Letters

Personal Experiences

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483 Observations There was inadequate method validation specificity

data to demonstrate that each method was capable of

distinguishing the active ingredient from its impurities

and degradation products.

Specificity studies did not include the minimum stress

conditions of acid and base hydrolysis, oxidation,

thermal degradation and photolysis, degradation

schematic for the active ingredient that identifies the

major degradation products

was not included for each product.502009

FDA Waning Letter

On addition to the example of modifying both compendial methods and customer supplied methods, we also observed the use of unvalidated in-house methods as well as unvalidated

modifications to in-house methods.A statement indicating that the method has not been

validated in the particular formulation was included in the certificate of analysis for…use of this statement does not absolve…from using valid, accurate, and

reproducible methods. (June 2000)

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FDA Systems Based Inspection:Laboratory System

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Method Validation

13%Training/Qual.

4%

Stability Program 21%

InadequateRecords

27%

Controls. General35%

Feb – July 2002: 212 Inspections (US)

* Reference: Albinus D’ Sa, FDA, CDER Office of Compliance, from AAPS, Nov. 2002 presentation.2009

ICH Update:

2009 53

A Unique Approach

International Conference on Harmonisation (ICH) was created in 1990

Agreement between the EU, Japan and the USA to harmonize different regional requirements for registration of pharmaceutical drug products

Unique because joint effort by regulators and associated pharmaceutical industry trade associations

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ICH Objectives

Identification and elimination of the need to duplicate studies to meet different regulatory requirements

More efficient use of resources in the R&D process, as a consequence

Quicker access for patients to safe and effective new medicines

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Working Groups

SAFETY EFFICACY

QUALITY MULTIDISCIPLINARY

STEERING COMMITTEEEndorses topics, guidelines and monitors progress

2009 56

Related Site

www.fda.gov

www.fda.gov/cder/

www.waters.com

www.usp.org

www.ich.org

www.aoac.org

www.pharmweb.net

572009