Method validation for drug substances and drug product _remodified_2014

Validation of Validation of

Analytical Method Analytical Method

for Drug Substances for Drug Substances

& Drug Products& Drug ProductsDr. R.Badmanaban., M.Pharm., M.D(A.M).,PhD.,

Associate Professor, Head of Dept - Pharmacognosy Shri Sarvajanik Pharmacy college - Mehsana-384001

www.sspcmsn.org Email: [email protected]

Definition:

Method validation is the process of proving that ananalytical method is acceptable for its intended purposes.

METHOD VALIDATION = ERROR ASSESSMENT

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

Analytical Method Validation

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

International regulatory bodies and their guidelines on different aspects of QA

Chromatography todayChromatography todayMore than sixty variants of the technique have been developed.HPLC, GC, SFC, and CE are the most frequently used.

HPLC: almost universal wide range of equipment and columns is commercially available well-understood separation mechanisms sensitive, specific, selective, precise and robust, Rugged,

accurate. easy to maintain instrumentation flexible in optimizing separations More efficient than some of the separation techniques

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Shri Sarvajanik Pharmacy college

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Liquid Chromatography (HPLC) instrumentLiquid Chromatography (HPLC) instrument

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The Chromatographic The Chromatographic Process – Theoretical Process – Theoretical ConsiderationsConsiderations

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Elution in Column ChromatographyElution in Column Chromatography

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MobileMobilephasephase

StationaryStationaryphasephase

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Intermolecular InteractionsIntermolecular Interactions

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ChromatogramChromatogram

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Retention RelationshipsRetention Relationships

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Validation (4M)

Man

Machine

Material

Method

Why Method Validation is Important

?Analytical Method

Validation

Why Method Validation is Important?

1. Develops confidence in using the method & Proof that

Method is suitable for its intended purpose,

The purpose of analytical measurement is to get consistent,

reliable and accurate data.

Incorrect measurement results can lead to tremendous

costs.

2.2. Regulatory requirementRegulatory requirement, Equal importance for those

working in a regulated and in an accredited environment.

U.S. FDA, ISO etc.

When to be validated

?


When to be validated?

Partial validation after development of method.

Complete validation after manufacturing formula is finalized.

Which methods are to be validated ?

Compendia: Pharmacopoeia method

Verification of suitability of method

Non compendia methods: Laboratory developed methods.

Pharmacopoeias methods used outside its scope.

Validation: Prior Considerations

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 and approved standard test procedure and proper approved protocol with pre-established acceptance criteria

Validation Activity Including the Complete Analytical

Procedure

Sampling

Sample Preparation

Analysis

Data Evaluation Reporting

Validation Step

Define the application, purpose and scope of

the method.

Analytes? Concentration?

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 Procedures for executing the method in

routine analysis.

Document validation experiments and results in the

validation report.

Validation Step

Verification vs. Validation

Compendial vs. Non-compendial Methods

Compendial methods-Verification

Regulatory analytical procedure in USP/NF

Non- compendial methods-Validation

Alternative analytical procedure proposed by the

applicant for use instead of the regulatory analytical

procedure

MethodValidation

MethodTransfer

MethodDevelopment

Approved

BACKGROUND-LAB METHOD FLOWBACKGROUND-LAB METHOD FLOW

ICH/USP Validation Requirements

Precision

Repeatability

Intermediate Precision

Reproducibility

Limit of Detection

Limit of Quantitation

Robustness

Specificity

System

suitability

Linearity

Range

Accuracy

Validation Parameters

ImpuritiesSpecificityLinearity and RangeAccuracy PrecisionRobustnessLOD & LOQs

Dissolution

SpecificityLinearity and RangeAccuracy PrecisionRobustness

Assay / CU

SpecificityLinearity and RangeAccuracy PrecisionRobustness

Specificity/Selectivity

Ability of an

analytical method

to measure the

analyte free from

interference due

to other

components.

Selectivity

Bias

Specificity: ICH/USPThe ability to measure accurately

and specifically the analyte in the

presence of components that may

be expected to be present in the

matrix

The degree of interference

Active Ingredients

Excipients

Impurities (synthetic

precursors, enantiomers)

Degradation Products

Placebo Ingredients


Continue....

. Combination of 2 or more analytical procedures may be required to

achieve necessary level of discrimination

. Stability indicating analytical methods should always be specific.

. Analysts should ascertain whether the peaks within a sample

chromatogram are pure or consist of more than one compound.

Therefore should know how many compounds are in the sample or

use procedures to detect peak purity


Specificity: Impurities Assay

Chromatographic MethodsDemonstrate Resolution

Impurities/Degradants Available Spike with impurities/degradants

Show resolution and a lack of interference

Impurities/Degradants Not AvailableStress Samples

For assay, Stressed and Unstressed Samples should be compared.

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SelectivityAbility 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

Selectivity: 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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2009


Linearity

Ability of an assay to elicit

a direct and proportional

response to changes in

analyte concentration.

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2009

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 >

0.999

Requires a minimum of 6 concentration levels

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2009

RangeThe interval between the upper and

lower concentrations of analyte in

the sample that have been

demonstrate to have a suitable level

of precision, accuracy, and linearity.

Range

Normally derived from Linearity studies.

Established by confirming that the

method provides acceptable degree of

linearity, accuracy, and precision.

Specific range dependent upon intended

application of the procedure.

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

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Accuracy

Closeness of the test

results obtained by

the method to the

true value.

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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2009

Accuracy Data Set (1 of 3)

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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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Accuracy Vs Precision

Inaccurate &imprecise

Inaccurate butprecise

Accurate butimprecise

Precision… Considered at 3 Levels

Repeatability

Intermediate

Precision

Reproducibility

Repeatabilit

y

Express the precision under the same

operating conditions over a short interval

of time.

Also referred to as Intra-assay precision


Express within-laboratory

variations.

Expressed in terms of

standard deviation, relative

standard deviation

(coefficient of variation) and

confidence interval.

Known as part of

Ruggedness in USP

(Different Analysts, Different

Laboratories, Different

Instruments, Different

Reagents, Different Days)

Depends on the

circumstances under

which the procedure

is intended to be

used.

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

Definition: Ability to reproduce data

within the predefined precision

Determination: SD, RSD and confidence

interval

Repeatability test at two different labs.

Reproducibility

Reproducibility Study

Lab 1 Lab 2 Lab 3

Day 1 Day 2 Day 1 Day 2 Day 1 Day 2

Analyst1

Analyst2

Analyst1

Analyst 2

Analyst 1

Analyst 2

3 Preps

3 Preps 3 Preps 3 Preps

3 Preps 3 Preps

Lowest amount of analyte

in a sample that can be

detected but not

necessarily quantitated.

Estimated by Signal to

Noise Ratio of 3:1.

Detection Limit (DL)

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.

Quantitation Limit (QL)

LOD, LOQ and SNR

Limit of Quantitation (LOQ)

Limit of Detection (LOD)

Signal to Noise Ratio (SNR)

noise

Peak ALOD

Peak BLOQ

Baseline

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

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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

2009


Definition: Capacity to remain

unaffected by small & deliberate

variations in method parameters

Determination: Comparison results

under differing conditions with precision

under normal conditions

Variations may include: stability of

analytical solution, variation of pH in

a mobile phase, different column

(lot/supplier), temperature, flow rate.

Robustnes

s

Robustness Variations

All Assays

HPLC Assays

GC Assays

-Sample Prep Manipulation

-Extraction Time

-Mobile Phase Composition

-Different Columns

-Temperature

-Flow Rate

-Different Columns

-Temperature

-Flow Rate

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 %RSD

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2009

Solutes may readily decompose prior to chromatographicinvestigations e.g. during sample preparation, extraction,cleanup, phase transfer or storage of prepared vials(refrigerators or automatic sampler).Method development should investigate the stability of theAnalytes AND standards.

Stability of analytical solution

Stability of analytical solution

Solution stability •Stability of the samples being analyzed in a sample

solution. e.g. 1 – 48 hours using a single solution.•should be determined by replicate analysis of the

samplesolution.

The checking of a system, before or during analysis of unknowns, to ensure system performance.

“No sample analysis is acceptable unless the requirements for system suitability have been met.” (USP Chapter 621)

Plate Count, Tailing, Resolution

Determination of reproducibility (%RSD)

For %RSD < 2.0%, Five replicates

System Suitability "Sample“ - A mixture of main components andexpected by-products utilized to determine system suitability

“Whenever There is a Significant change in Equipment or ReagentsSystem Suitability Testing Should be Performed” (USP Chapter 621)

SYSTEM SUITABILITYSYSTEM SUITABILITY

Confuse of Precision Terms

Repeatabili

ty

Intermediat

e Precision

Reproducibili

ty

Ruggedness

Robustness

Precision Terms

Instrument Precision

Repeatability


Reproducibility

Ruggedness

Robustness

- 6 Standard

Injections

- One Analysis (6

preps)

- Two Analyses

- Two different Lab.

- Many Variables

- Intentional

Changes

Change in the analytical procedure, drug substance,

drug product, the changes, may necessitate

revalidation of the analytical procedures.

“The degree of revalidation depends on the nature of

the change.”

“FDA intends to provide guidance in the future on post-

approval changes in analytical procedures.”Revalidation should accompanyformulation changes (new samples with new compounds or new matrices)manufacturing batch changesnew analysts with different skills,new instruments with different characteristics,new location with different environmental conditions,new chemicals and/or reference standards andmodification of analytical parameters.

Revalidation

Validation ReportValidation Report

Objective and scope of the method (applicability, type).Summary of methodology.

Type of compounds and matrix.All chemicals, reagents, reference standards, QC samples with purity, grade, their source or detailed instructions on their preparation. Procedures for quality checks of standards and chemicals used.Method parameters.Critical parameters taken from robustness testing.Listing of equipment and its functional and performance requirements, e.g., cell dimensions, baseline noise and column temperature range.Detailed conditions on conduct of experiments, including sample preparation Statistical procedures and representative calculations. Procedures for QC in routine analyses, e.g., system suitability tests. Representative plots, e.g., chromatograms, spectra and calibration curves. Method acceptance limit performance data and expected uncertainty of measurement results.Criteria for revalidation.The person's) who developed and validated the method.References (if any).


How do we Know the expectations of the FDA?

FDA Form 483

FDA Warning Letters

Personal Experiences

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2009

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.

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2009

FDA Warning Letter

On addition to an example of modifying both compendia methods and customer supplied methods, we also observed the use of invalidated 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 2009)67

2009

General requirementsGeneral requirements

Qualified and calibrated instruments

Documented methods

Reliable reference standards

Qualified analysts

Sample integrity

Change control (e.g., synthesis,)

Analytical methods should be used within GMP and GLP environments, and must be developed using the protocols and acceptance criteria set out in the ICH guidelines Q2 (R1)


Related Site

www.fda.gov

www.fda.gov/cder/

www.waters.com

www.usp.org

www.ich.org

www.aoac.org

www.pharmweb.net

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2009

Thank you

Documents

Method validation for drug substances and drug product _remodified_2014