international analytical methodology

Quality by Design Highlights

Analytical Method Guidelines, Transfer, System Suitability, Traceability, and Variability Minimization

By

Satendra Kumar Vishwakarma, PhD

Declaration on Outlines

The materials/ contents presented in the following slides , collected

from various sources, reflect general information and minimum

requirements for conducting analytical activities in a qualified

laboratory system for regulatory submissions.

General Outlines are not complete Reference Guidelines

The readers are further requested to consult First Corporate Method

Guidelines and then other appropriate Regulatory Web Sites for

additional details and verification of outline contents.

Ж♦Ж♦Ж♦Ж Thank You Ж♦Ж♦Ж♦Ж

Outlines of Presentation

R&D Analytical CMC (Activity Classification and Responsibilities)

Departmental Analytical Suitability

Origin of Assay Method and Classifications

Definition and Determination of Selectivity and Specificity

International Validation Terms and Terminology

Data Elements for Verification

Methods for Cleaning Validation

Chromatographic Adjustments and Changes in HPLC

Technical Method Transfer Criteria & Concept

International System Suitability Requirements

Impurities/ Degradants in Drug Products/ Drug Substances

Variability and Minimization by Quality by Design in Laboratory

Quick Web Link References

Compliant Analytical CMC Activity

API, Excipient

Evaluation &

Control

Pre-/ Post-

Formulation

Support

Compatibility

Studies

Support

In-Process

Studies

Support

Specification

Safety

Evaluation

Analytical Procedure &

Method Development

API, Product, & Packaging: Safety, Quality & Performance Testing Chemistry of Drug Molecule: Characterization of Reference & Raw Material and Control*

Chemistry of Excipients: Characterization and Variation Control* for intended functions

Chemistry of Drug Interaction: Preformulation/ Formulation Compatibility

Drug Analysis: Method Development, Validation, Verification, and Transfer

Stress Degradation and Stability Studies

Identification/ Quantitation of Known and Unknown Impurities

Acceptance Criteria/ Specification Development

Method Life Cycle Management

Bioequivalence: Verification of Product Quality and Integrity Studies

Evaluation: Packaging, Drug Substances/ Drug Product Intrinsic Stability Testing

Process Verification: Process Validation Batches Data Development

Regulatory and Guidance: Compliance Data/ Reports and Maintenance

* Worldwide Historical data on pharma products indicates that poor characterization and control

on raw materials accounts to Long- term Manufacturing and Product Performance Issues.

QA &

Regulatory

Compliance

The Old Way of Product / Substance Analysis – Lab:

Off-line: Current Laboratory practices are destructive and non-destructive (regulatory

disadvantage).

The New Way of Product / Substance Analysis – PAT:

At-line:In production area, analysis performed during production close to the

manufacturing process.

On-line: Analysis on diverted stream connected to process.

In-line: Process stream disturbed.

Non-invasive: Sensor not in contact with material, process not disturbed (regulatory

advantages).

Goals of both (Lab and PAT Way): Provide information about the sample of interest.

Perspective different:

Lab Way: Time dependence not critical for technical reasons.

PAT Way: Typically used to control processes through feedback in real time or to diagnose

and explain process anomalies.

This dictates differences between the Two Ways with regard to Sampling and Rate of Data Collection

State of Process Product Analysis

Process Product QualityProcess Analysis – Sources of Error and Variability

PAT

Current Quality By Lab Future Quality By Design

Off-Line At-Line / By-Line In-Line / On-line

Product Oriented Process Oriented

Drug Release, Impurities Blending, Homogeneity, Filling, Granulation, Drying, Coating, Screening

Manual Sampling

Transportation Sample Preparation

Slow Response

Quality Not Controlled

High Manufacturing Cost

Manual Sampling

Sample Preparation / Minimum Preparation

Rapid Response

Quality Controlled

Low Product Cost

Sample At Interface

Direct Analysis

Immediate Response

Rapid Quality Controlled

Low Manufacturing Cost

Excipients Choice in

Dosage Forms

Physicochemical

Properties of Drug

Polymorphic / Forms

Hydrates

Heat & moisture sensitive

Poorly Soluble

Poorly absorbable

Poorly Stable in vivo

Physicochemical

Properties of Excipients

Physically Stable

(Polymorphic / Forms

Hydrates)

Hygroscopic

Chemically Stable

Compatible with drug

Rheology Flow

Route of Administration

Oral

Injection

Pulmonary

Transdermal

Buccal

Rectal/Vaginal

Desired Release

Characteristics

Immediate release

Sustained Release

Modified Release

e.g. enteric

Delivered Dose

of Drug

High Dose

Low Dose

Manufacturing

Process Requirement

Direct compression

Wet Granulation

Fluid Bed Coating/

Granulation

Spray Drying

Other novel

processes

Direct Liquid

Formulation

Filtration & Fillings

Excipients in Product QualitySelection and Characterization

Dosage Form Development Chart

Active Drug

Suspension

Solution

Intrinsic

Dissolution

Dissociation

Constant pKa

Intrinsic

Solubility

Suppositories Topicals

pH Effect Co-solvents

Capsule

Salts Saturated

Solubility IV Injection

Solution

Other

Delivery

System

Other

Dosage Forms

Tablets

Stability

Tonic

Adjustment

Tonic

Adjustment

Excipient Compatibility

PEG 400 + 5%

H2O + Glycerin

pH?

Analytical Quality-by-DesignMolecular Properties and Impurities Identification

Method Development

•Spectroscopy, Hyphenated, Orthogonal Techniques (Process Tools) in DS/DP Impurities Identification.

•Molecule Properties Identification and Characterization like Polymorphs and Polarity Screening by HPLC.

•Application of Multiple or Suitable Detectors.

•Selectivity/ Identification and Application of On-line Analyzers.

•Application of Target Methods for Particular Impurities or Enantiomers.

Formulation Development

• Physiochemical Properties Identification of Formulation Components.

• Phases Interaction and Evaluation at Rapid/ Accelerated Environmental Conditions.

• Application of on-line Laboratory based Analyses and Chemometrics Knowledge.

• Finger Print Mapping of Formulation Components.

•Re-optimization of Methods against Re-optimized Formulations.

Process

Understanding

•Monitor the identified Process Variable Parameters.

•Control the Impurities by Controlling the Variable Parameters.

•Direct Application of on-line Laboratory based Knowledge.

Control

Methods

• Integral to Product Specifications.

• Integral to Process Controls.

•Optimization for Ruggedness.

Analytical Quality-by-DesignFT-IR and HPLC Comparison

Features FT- IR HPLC

Sample Preparation Non-Destructive Pipetting, Crushing &

Dissolving

Time per Analysis Typically 30-90 seconds Typically 20-40 minutes

Solvent Required No Yes

Chemical Information Active Concentration/

Uniformity

Active Concentration/

Uniformity

Physical information Yes No

Surface Analysis Yes in reflectance mode No

Moisture Content Yes No

Low Concentration Impurities No Yes

QbR/ Regulatory Compliance Yes Yes/ No

Smart Analytical NIR Spectral Range

Cosmic Gamma

X-r

ay U

V IR Micro UHF

Sh

ort

Me

diu

m

Lo

ng

Ultra violetInfrared

Near Mid Far

1 400 750 2500 16000 1000000 nm

Radio

Vis

Scan NIR spectrum range ≈ 780nm - 2526nm (12820-3959cm‾¹) non-invasively and non- intrusively. Scan in the

range of 1100nm – 2300nm @ 2nm intervals. Spectra generated is average of 100 NIR scans

Elemental Appearance Constituent Assay Functional Group Analysis

Departmental Analytical Suitability

Suitability of Analytical Instrument & Support SystemCurrent status of Qualification, Calibration, and approved documentation.

Suitability of Required MaterialsQualified reference standards, APIs, reagents, matrix/ placebo.

Suitability of Analytical ChemistApproved status of qualification, training, and GLP-GMP experience.

Suitability of Method Validation DocumentationA well written analytical procedure and approved protocol with pre-established

acceptance criteria.

Suitability of Method/ Method Transfer DocumentationA written protocol, defined responsibilities, identified & approved analytical

procedure, statistical analysis, pre-defined acceptance criteria for each

performance parameters, time frame with solutions stability time and

suitability parameters.

Suitability of Data, Method Audit, and Deficiency DocumentationChemist authorization, data audit trail & control, method audit trail, deviation/

Investigation, change control and records management.

Regulatory Conditions form Method Development to Method Applicability

Assay Method Classifications

Classification of Analytical Assay Methods & Methods to be Validated

Method Class Definition by Performance Characteristics

Class 1

(Identity)

Analytical methods for Quantification (Identification) of major

components in drug substances (active ingredients), drug

products (finished products), support vehicles (preservatives

and excipients).

Class 2

(Detect &

Quantitate)

Analytical methods for Determination (Identification,

Quantification , Limit ) of known and unknown impurities in

bulk drug substances (active ingredients), drug products

(finished products), support vehicles (preservatives and

excipients).

Class 3

(Concentration)

Analytical methods for Determination of Performance

Characteristics – Drug Release ( e.g. Dissolution of Drug

Products).

Class 4

(Characteristics)

Identification tests – Physical, Chemical limit tests, FTIR, Chiral

Test, & Spectroscopy.

Stages of Continuous Verification & Validation of Pharmaceutical HPLC Method(from Drug Discovery to Early Drug Development to IND to NDA)

Analytical Validation

Parameters (v)

Research & Method Development (RMD) Technology

Early Drug

Development

Formulation Development (FD) Mfg Method

Transfer

& SubmissionPre-FD-Phase-1 FD-Phase-2 Final FD-Phase-3

RMD-1 RMD-2 RMD-3 RMD-4 RMD-5

Accuracy … v v v v

Precision

Injection Repeatability v v v v v

Impurity Reproducibility … … v v v

Assay Intermediate … v v v v

Specificity/ Forced Deg … v v v v

Detection Limit … … v v v

Quantitation Limit … v v v v

Linearity v v v v v

Analytical Range v v v v v

Solution Stability … … v v v

Robustness … … v v v

Evolution of Regulatory Method

Validation/ Verification Terminology

Assay Parameters Basic Description of Analytical Performance Parameters

Accuracy Evaluate the closeness of “Measured” value & „True‟ value.

Imprecision Evaluate the variability in replicate measurements (intra- and

inter-assay).

Specificity

(Selectivity)*

Evaluate the ability of the method to measure all other

components without reacting with other related substances.

Analytical Range Establish analytical range of Active and Impurities over which the

method shows acceptable performance.

Reportable Range Establish range of reportable results over which the method

is validated (may exceed analytical range when samples are

diluted or concentrated).

Sample Stability Evaluate stability of reference standard, impurity solution, and

sample matrix under conditions that mimic the study conditions.

Quality of Standards Evaluate the quality of calibrators/ standards.

Calibration/ Standard

Curve

Evaluate relationship between known quantities (concentration) of

reference standards in artificial or true matrix and measured

instrument response.

* See next slide for Definition and Significance of Specificity

Specificity (Selectivity)**

The analytical performance and ability of a method to measure accurately

and specifically the analyte in the presence of complex components (Active

Ingredients, Degradation Products, Placebo Ingredients, Impurities) that may

cause a degree of interference. This requires separation and characterization.

Separation Resolution (Determination of separation between peaks), Plate Count

(Determination of a systems efficiency), Tailing Factor (Calculation reference

peak shape).

Characterization (PDA/DAD) Peak Purity Test (angle and plots) is necessary to demonstrate that the

analyte chromatographic peak is not attributed to more than one components.

** See slide on “Stress or Forced Degradation” & its relation with Method Specificity

Definition of Specificity/ Selectivity

Determination of Specificity/ Selectivity

Qualitative Identification Tests

Demonstrate ability to select between compounds of closely related structure

and confirm positive and negative results.

Assay

Demonstrate that the results are unaffected by spiked impurities or excipients.

Impurities If the Impurities are Available

Spike the drug product/ drug substance with impurities and demonstrate

appropriate accuracy and precision. This demonstrate that assay is

unaffected by the presence of spiked materials (impurities and/or excipients).

Compare the results on unspiked assay samples.

If the Impurities are Not Available**

Compare test results to a second well-characterized procedure. For Assay,

compare the two results. For Impurity Tests, compare impurity profiles Peak

Purity Test ( by diode array detector or by mass spectrometer). Compare the

results obtained under stress (forced degradation) conditions samples.

** See slide on “Stress or Forced Degradation” & its relation with Method Specificity

Methods Under Guidelines

Validation of New Test Methods (Also known as Complete Method Validation)

All New Non-compendial or Alternative Test methods will be evaluated as per ICH/FDA

Critical Raw Material and Components Tests

Process Validation and In-Process Tests

All GMP Lot Release Tests

1. Potency and Purity

2. Stability Assays for Approved Products

3. Other Safety Tests including Sterility and Pyrogenicity

Verification of Pharmacopeia Methods (Also known as Partial Method Validation)

All USP/EP Compendial Tests

In-house Test Methods already used for Approved Products and Not further Modified

System Suitability must be Demonstrated

1. Ensure Test System Working Properly when Run Performed

2. Needs Equipment Qualification and Periodic Calibration

3. Needs Demonstration of Analyst‟s Competence

4. Needs Reliable Reference Standards and Reagents

5. Test perform for specificity, intermediate Precision and sample solution stability

Qualification of Test Methods

Comparability and Characterization Tests

Stability Tests, Pre-NDA

In-Process and Final Quality Tests during Development

Some Clinical Test depending on Clinical Phase

International Analytical Methods

Analytical Methods and their Analytical Performance Characteristics

Validation Parameters FDA ICH USP EP JP

Accuracy (Trueness) Yes Yes Yes * Yes

Precision (Repeatability / Intra-Assay)

Precision (Reproducibility / Labs)

Precision (Intermediate / Ruggedness)

Yes

Yes

Yes

Yes

Yes

Yes

Yes

Yes

Yes

*

*

*

Yes

Yes

Yes

Specificity Yes Yes Yes * Yes

Detection Limit Yes Yes Yes * Yes

Quantitation Limit (= Detection + Concentration) Yes Yes Yes * Yes

Linearity Yes Yes Yes * Yes

Analytical Range Yes Yes Yes * Yes

Reportable Range Yes Yes Yes * Yes

Robustness Yes Yes Yes * Yes

Analytical Solution Stability Yes Yes Yes * Yes

Term Precision means 4Rs - Repeatability, Reproducibility, Ruggedness and Robustness

Data Elements for Validation

Table I: Data Elements required for Validation in a given Method Class

Analytical

Performance

Characteristics

Class 1 Class 2 Class 3 Class 4

Quantitative Limit Tests

Accuracy Yes Yes MBR MBR No

Precision Yes Yes No Yes No

Specificity (Selectivity) Yes Yes Yes MBR Yes

Detection Limit No No Yes MBR No

Quantitation Limit No Yes No MBR No

Linearity Yes Yes No MBR No

Range Yes Yes MBR MBR No

Ruggedness Yes Yes Yes Yes Yes

MBR = May Be Required, depending upon nature of the specific tests

Data Elements for Verification of

DRUG SUBSTANCESTable II: Data Elements required for Verification of Drug Substances and Excipients

Analytical

Techniques



HPLC/GC Precision Precision

Specificity

Quantitation

Specificity

Detection

-

-

-

Specificity

Spectrophotometric/

Colorimetric

Precision Precision

Quantitation

Specificity

Detection

- Specificity

Titrimetric Precision Precision - - -

TLC - Specificity

Quantitation

Specificity

Detection

- Specificity

Electrophoresis - Specificity

Quantitation

Specificity

Detection

- Specificity

DRUG PRODUCTSTable III: Data Elements required for Verification of Dosage Forms (Products)

Analytical

Techniques



HPLC/GC Precision

Specificity

Linearity

Range

Precision

Specificity

Quantitation

Specificity

Detection

Precision Specificity

Spectrophotometric/

Colorimetric

Precision

Specificity

Range

Precision

Quantitation

Specificity

Detection

Precision Specificity

Titrimetric Precision

Linearity

Range

Precision - - -

TLC - Specificity

Quantitation

Specificity

Detection

- Specificity

Electrophoresis - Specificity

Quantitation

Specificity

Detection

- Specificity

Data Elements for Verification of

International Validation ParametersCommon Analytical Methods and their Assay Validation Parameters

Validation Parameters

(Analytical Characteristics)

M E T H O D S

ID Impurities Cleaning Assay Specific

TestQuantitation Limits

Accuracy (Trueness) N Y N (Y) Y Y Y1

Precision – Repeatability

Precision – Intermediate

Precision – Reproducibility

N

N

Y

Y

Y3

Y

N

N

N

Y

N

Y

Y

Y3

Y

Y1

Y1

Y1

Specificity (Selectivity/ Sensitivity) Y2 Y Y Y Y Y

Detection Limit N N (Y) Y Y N N

Quantitation Limit N Y N Y N N

Linearity N Y N Y Y N

Range N Y N (Y) Y Y N

Robustness Y Y N N (Y) Y Y1

Surface Recovery N N N Y N N

Stability Indicating N Y N N Y N

Solution Stability (Standard/Sample) N Y Y Y Y Y

Reference Standard Evaluation Y Y Y Y Y Y

N = Signifies that this characteristics is not normally evaluated. Y = Signifies that this characteristics is normally

evaluated. N(Y) = May be needed in some cases. 1 = May not be needed in some cases. 2 = Lack of specificity for

an analytical procedure may be compensated for by addition of second analytical procedure. 3 = In cases where

reproducibility has been performed, intermediate precision is not needed.

Method Re/Validation Parameters

Requirement for Revalidation of Analytical Methods

Accuracy Influence of formulation components

Precision Influence of formulation and sample preparation

Specificity Presence of new API(s) and impurities / degradants /

formulation components

LOD/LOQ Test concentrations of API(s) versus FPP)

Range* Test concentrations of API(s) versus FPP

Robustness Change of column material, column parameters, solvents

Partial/ Revalidation Required by ICH Q2A under Circumstances

Changes in the synthesis of the drug substance

Changes in the composition of the finished product

Changes in the analytical procedure or method is modified and modifications

are outside original scope of the method. (e.g. robustness)

Change in analytical instrument conditions for which the method has been

validated (e.g. Instrument with different characteristics)

* Revalidation is necessary, if ranges covered during validation of API-methods

are different from those of the FPP-methods (different test concentrations).

Methods for Cleaning Validation

Procedure Assay and Related Substances used in Stability

Studies of API and FPP

Specificity Samples taken from a cleaning assessment

Linearity Response (from 50% of the cleaning limit to 10x

this concentration; R2 ≥ 0.9900)

Precision Repeatability Precision (RSD ≤ 5%)Intermediate Precision [Ruggedness (USP)]Reproducibility Precision

Limits of Detection N(Y) Desirable to set in some cases

Limit of Quantitation Acceptance Criteria

Accuracy or

Recovery

Rinsate (≥ 80%), Swabs (≥ 90%), and process

surface (≥ 70%)

Range Lowest level is at least 2x higher than LOQ

Multiple Adjustments / Changes? Regulation may require Additional-/Re-Validation

HPLC Parameters Specification Comments

pH of Aq. Mobile Phase Within ± 0.2 units (value or range) …

Buffer Salt Concentration ± 10% Provide pH variation is met

Ratio of Mobile Phase

Components

Components specified at 50% or

less: ± 30% or ± 2%

Whichever is larger. Change in any

component can not exceed ± 10% Absolute,

nor any reduced to Zero.

UV-Visible Detector

Wavelength

No deviations A validated procedure must be used to verify

that error in the detector wavelength setting

is, at most, ± 3.0 nm.

Column Length ± 70% …

Column Inner Diameter ± 25% …

Flow Rate ± 50% …

Injection Volume Reduced as far as consistent with

accepted Precision and Detection

Limits

Increased to as much as 2X volume specified

as long as there are no adverse

chromatographic effects.

Particle Size Reduced by as much as 50% …

Column Temperature ± 10% °C Recommended to improve control and

reproducibility of RT

(Login Required) http://www.uspnf.com/uspnf/pub/index?usp=31&nf=26&s=0

USP Adjustments/Changes in LC

http://www.uspnf.com/uspnf/pub/index?usp=31&nf=26&s=0

Multiple Adjustments / Changes? Regulation may require Additional-/Re-Validation

HPLC Parameters Specification and Comments

pH of Aqueous part of

Mobile Phase

May vary by ± 0.2 pH unless otherwise stated in the monograph, or ± 1.0 pH for

neutral substances.

Column Temperature Can be adjusted by ± 5 °C.

Ratio of Mobile Phase

Components

The amount of minor solvent component can be adjusted by ± 30% relative (or ±

2% absolute).

UV-Visible Detector

Wavelength

No Adjustment is permitted.

Buffer Salt Concentration Concentration of salts in the buffer component of mobile phase: ± 10%.

Flow Rate ± 50%

Injection Volume May be decreased provided detection & repeatability are satisfactory.

Stationary phase Column Length: ± 70%, Column Int. Diameter: ± 25%, Particle Size: max - 50%, no

increase permitted. Flow Rate correction required.

Gradient Elution Mobile Phase composition not permitted

The percentage variation of Ion Pair Reagent were not found in either USP or EP

EP Adjustments/Changes in LC

Technical Method Transfer

R&D Analytical Method Transfer to Regulated Quality Control Laboratory

Procedure and Acceptance Criteria Driven Technical Protocol to fulfill proficiency/

reliability checks for Precision, Accuracy and Ruggedness parameters

Do not change Validated Chromatographic Variable Parameters

Ionic strength in mobile phase

Solvent strength or ratio in mobile phase

Final pH of mobile phase

Temperature (column, vial, mobile phase)

Flow rate (isocratic / gradient ratio and time)

Sample diluent / extraction /sonication time

Test solution stability

Injection volume

Specified analytical column (No equivalent terminology)

Mode of detection or Wavelength of detection

Measure Chromatographic Parameters

Analytical system suitability parameters and injection frequency

Response (area/amount) for repeatability & reproducibility

Retention time

Selectivity and/or resolution

Method Transfer Acknowledgement/ Documentation/ Return with Feedback

Document the acceptance criteria with pertinent observations/ Return to

originating laboratory on failure for further optimization.

Evolution of Stability Indicating Method

DOE for Development

Screening

DOE for Qualification

Optimization

Pre-validation

Validation

Implementation

Re-v

ali

da

tio

n

Method Transfer

Characterization Identification

Re

-va

lida

tion

QC/ Stability

Problem

Formulation

Problem

Information

Purity & Stability Indicating Method

A stability method transfer must be Stability-Indicating Assay

Method is a validated quantitative analytical procedure that can

detect the changes with time the pertinent properties of the drug

substance and drug product.

A stability-indicating assay accurately measures the active

ingredients, without interference from degradation products, process

impurities, excipients, or other potential impurities.

A stability indicating methods are developed, optimized and adapted

according to the ICH principles for forced degradation studies.

A non-stability indicating analytical procedure can be used for

release testing, then an analytical procedure capable of qualitatively

and quantitatively monitoring the impurities, including degradation

products, is required to complement it.

Assay analytical procedures for stability studies should be purity and

stability-indicating, unless scientifically justified.

Method Transfer Testing Criteria

Method Transfer Testing Criteria involves either for

Verification of Method (under actual conditions – Validation requirements),

Qualification of Method (performing tests– Specification requirements) or

Comparison of Method (evaluation of test solutions – Confirmation requirements)

Best Laboratory Practices/ Guidelines for Qualification Transfer Criteria

Method Class Class 1

Assay

Class 2 Class 3

Performance

Class 4

ID TestQuantitative Limit

Parameters

Accuracy Y Y N N N

Precision Y Y N Y N

Specificity Y, 1 Y1 Y, 2 N Y, 2

Quantitation NR Y N N N

Linearity Y Y N N N

1 Tested with respect to Sample Diluent.

2 Positive response for analyte presence. No response required for Blank.

Y Verifying laboratory needs to perform tests.

NR Not required.

N Testing not suggested.

Method Transfer Testing CriteriaMethod Transfer Testing Criteria involves either for

Verification of Method (under actual conditions – Validation requirements),

Qualification of Method (performing tests– Specification requirements) or

Comparison of Method (evaluation of test solutions – Confirmation requirements)

Best Laboratory Practices/ Current Validation / Revalidation Guidelines for

Comparative Testing Criteria: Originating (O) & Receiving (R) Laboratories

Method Class Class 1

Assay

Class 2 Class 3

Performance

Class 4

ID TestQuantitative Limit

Comparative

LaboratoryO R O R O R O R O R

Parameters

Accuracy N N N N N N

Currently

Not

Required

Currently

Not

Required

Precision Y Y Y Y N (Y) N (Y)

Specificity N Y N Y N Y

Detection N N N N N N

Quantitation N N N Y N N

Linearity N Y N Y N N

Range N N N N N N

Results Y Y N(Y)

Method Transfer Design Text

Typical Method Transfer Experimental Design and Acceptance criteria Example

Method Type Chemists Product Acceptance Criteria Comments

Assay 2 3 Batches in

Triplicate

A two one-sided t-

test with intersite

differences of ≤ 2% at

95% CI

Each chemist should use different

instrument & columns, independent

preparations. System suitability

criteria must be met.

Impurities and

Degradation

2 3 Batches in

Duplicate

(Triplicate if

done

together

with assay

For high levels, a two

one-sided t-test with

intersite differences

of 10% at 95% CI. For

low levels, criteria

are based on the

absolute difference

of mean ±25%

All samples age, homogeneity

packaging and storage should be the

same. All system suitability criteria

must be met. The LOQ should be

confirmed and chromatograms should

be compared with impurity profile. If

samples do not contain impurities

above reporting limit, then spiked

sample are recommended

Identification 1 3 Batches RT/ Spectral/

chemical results

Cleaning

Validation

1 Two spiked

samples –

one above

and one

below limit/

spec

Spiked levels should

not deviate from by

an amount 3 x the

validated standard

deviation of method,

or 10% of the spec,

which ever is greater

All system suitability criteria must be

met.. Cleaning is a limit test. Low and

high samples to confirm both positive

and negative information is required

Regulatory Analytical Suitability

Instrument Calibration & Computer System Validation

Pump Module Injector Module

Detector Module Temperature Module

Validationof

Chemist & Method

System Suitability

Testing & Criteria

Regulatory System Suitability

What‟s in System Suitability Criteria (Testing) System suitability testing is an integral part of many analytical procedures.

The test concept constitute equipment, electronics, analytical operations,

standards, and samples.

System suitability criteria are Not The Same as method performance criteria.

They usually provide a surrogate measure of the method performance criteria

(Retention Characteristics, Resolution, Tailing, etc.)

System suitability are limits to various tests and are designed to ensure the

adequate performance of analytical procedure (Repeatability of Injections).

System suitability test parameters to be established for a particular procedure

depend on the type of procedure being validated.

System suitability requirements should met before samples are analyzed.

System suitability must be performed for each study at the beginning and at the

end by each analyst, preferably at the middle of standard loop.

System Suitability may be needed to demonstrate / determine carryover and

confirm reagent purity by injecting blank (diluent or matrix).

Compliance with the system suitability criteria (sensitivity and selectivity) is

required throughout the chromatographic procedure.

USP & ICH System Suitability

USP<1225>System Suitability Parameters RequirementsCapacity factor (K‟) : ≥ 2.0

Peak Resolution (Rs) : > 2.0

Symmetry/ Tailing Factor (T) : ≤ 2.0

Theoretical Plates (N) : > 2000

Repeatability (Reproducibility), RSD : ≤ 2.0% (n≥5)

Repeatability (Reproducibility), RSD : ≥ 2.0% (n≥6)

Separation Factor (Relative Retention): Set Retention Time Window

ICH <Q2(R1)>Go to Respective Pharmacopoeias 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).

USP is the only document that comes the closest to

specific guidelines on System Suitability.

EP System Suitability Criteria

Suitability in terms of System Performance System Suitability Criteria are limits applied to various tests

designed to ensure the adequate performance of analytical

procedure.

Compliance with the system suitability criteria is required

throughout the chromatographic procedure.

Suitability in terms of Selectivity Resolution of two closely eluting peaks (critical pair): preferably

peaks of similar size or at least not saturating).

Peak-to-valley ratio (incomplete separation, peaks of very different

size)

“Similarity” or “Concordance” with a chromatogram supplied.

“Limit” Percent of individual impurity

“System Suitability” Signal-to-noise minimum 3 for the peak due

to impurity in reference solution.

Method =

Method Stability: System Suitability and Repeatiability Over Time

Stability of Analytical Solutions

Sample Solution Stability

Impurity-spiked Sample Solution Stability

Method Selectivity (Specificity & Sensitivity): The Sensitivity for an impurity can change depending on detector, lamp condition, mobile

phase composition and purity, column efficiency, etc.

Wavelength selection is often optimized for active ingredient but also should be considered

of impurities. Wavelength sensitivity should be demonstrated during stress testing studies.

Detection Limits (is estimated as 0.02% w/w by visual inspection of chromatogram) should

be evaluated for all significant potential impurities and degradation products.

Limit of Quantitation is estimated as 0.05% w/w and sample at this concentration included in

linearity (0.05% to 0.5% w/w) and in accuracy.

Significant impurities and degradation products include those that are specifically monitored

as part of Release Testing and Stability Testing.

Method Adaptability: Meets acceptance criteria of Specifications (limits), Robustness and Ruggedness

performance characteristics of intended applicability of analytical method.

Σ Stability & Selectivity∫

Organic Impurity and Degradant

Specified Impurity An impurity that is individually listed and qualified in product with a

specific acceptance criterion (can be identified or unidentified).

Impurity Definition (API Impurities) An impurity is any substance that is in the Drug Substance……..

that is not that Chemical Entity.

Conclusion: It may be: Known, Unknown, Specified, or Unspecified.

Degradant Definition (FPP Impurities) A degradant is an Impurity resulting from a Chemical Change of the

Drug Substance brought on by Manufacturing or Storage of a Drug

Product.

Conclusion: It may be: Known, Unknown, Specified, or Unspecified.

Reference Identification Procedure (API & FPP) An analytical procedure to differentiate and separate potential

impurities from matrix under a given conditions and specification.

Conclusion: It may be: Specificity, Sensitivity, Stability and Purity.

Quick Link: http://www.fda.gov/cber/gdlns/ichq3a.htm#att1

http://69.20.19.211/cder/guidance/2452fnl.htm#ATTACHMENT%20I

http://www.fda.gov/cber/gdlns/ichq3a.htm

http://69.20.19.211/cder/guidance/2452fnl.htm

Stress or Forced DegradationMethod Specificity through Forced Degradation Activities

ICH Q2 (R1)

Stress studies (e.g. products of acid and base hydrolysis, thermal degradation,

photolysis, oxidation) for the drug substance and for the active ingredient in the

drug product should be provided to demonstrate the specificity of the assay and

analytical procedures for Impurities.

Goals

To understand the drug substance stability.

To establish degradation pathways.

To validate the stability indicating power of the analytical procedures used.

To support the severe conditions that may be encountered during

distribution.

To generate typical degradation products which may be expected on stability

at sufficient levels to allow identification.

To avoid secondary degradation.

To get the target range is 5-20 % loss of active as judged by assay relative to

an un-degraded sample.

To look for purity and mass balance.

ICH Guidelines on Stress Testing

http://www.ich.org/cache/compo/276-254-1.html

Reference Subject Title

ICH Q1A(R2) Stability Testing of New Drug Substances and Products

ICH Q1B Stability Testing: Photostability Testing of New Drug Substances

and Products

ICH Q2(R2) Validation of Analytical Procedures: Text and Methodology

ICH Q3A(R2) Impurities in New Drug Substances

ICH Q3B(R2) Impurities in New Drug Products

Stress Testing – Forced Degradation (API)Stress studies elucidate intrinsic stability of the API and is part of the development strategy and is normally carried out under more severe conditions than those used for accelerated testing.Stress Testing – Forced Degradation (FPP)Studies undertaken to assess the effect of severe conditions on the FPP. Such studies include photostability testing and compatibility testing on APIs with each other in FDCs and API(s) with excipients during formulation development.

Stress or Forced Degradation






Impurities / Degradants From Stress

Stress conditions for Exploratory and

Definitive studies*

Drug Substance/

Product, %**

Impurity

Identification***

Peak Purity

Match****

Control Y Y Y

0.1 M HCl/ Temp/ Hrs Y Y Y

0.1M NaOH/ Temp/ Hrs Y Y Y

3% H2O2 / Temp/ Hrs Y Y Y

Humidity/ % RH/ Temp/Days Y Y Y

UV (Short and Long wavelength / Temp/

Days

Y Y Y

Light/ Lux/ Days Y Y Y

Thermal/ Temp/ Hrs+ Y Y Y

Transitional ion, Fe / Cu (Optional) … … …

Visit ICH Guidelines Website for Stress Stability Section for further Information

* Variable environmental conditions may be needed based on stability of molecule.

** Desirable to achieve realistic impurities/ degradation level 10 % to 30 % (NMT 50%) based

on active component mass balance.

*** Desirable to monitor impurity/ degradants at various wavelengths.

**** Drug Peak Purity Match is highly desirable (to indicate Peak Purity Indicating Method).+ Induced temperature should be below the melting point by 10°C to 20°C.

Source of Impurities / DegradantsPotential Impurities (degradants) Source: May be affecting Drug Stability

Impurities in starting materials

Products of over reaction or decomposition

Products of incomplete reaction and synthesis precursors

Impurities from the solvents of the reaction

Impurities from catalysts

Products of side reactions (synthesis by-products)

Degradation products (metabolites)

Residual solvents

Inorganic impurities

Impurities in excipients or reaction products of API with excipient(s) in formulation

Process Related impurities (e.g. Thermal Sterilization, Oxidative Environment)

Reaction products of API / Drug Product with immediate container / closure system

Enantiomeric forms as impurities*

Polymorph forms as impurities*

*(basically these are not impurities)

Impurities: Identification & Characterization

Impurities Identification: (Not exhaustive List) Organic Impurity

Each specified identified impurity

Each specified unidentified impurity

Any unspecified impurity with an acceptance criterion of not more than ()

the figure in the identification threshold [ICH Q3A/B(R)]

Total impurities (including extractable and leachable impurities)

Residual Solvents (By GC/GC-MS: Not covered in this presentation)

Inorganic Impurities (By compendial methods: Not covered in this presentation)

Impurities Characterization:

Impurity ID Chemical Name Molecular Structure Source/ Origin

Impurity A Chemical Name A Structure A Degradation and process

impurity

Impurity B Chemical Name B Structure B Process impurity

Impurity C Unknown Structure unknown

(RRT by HPLC)

Process impurity (levels do not

increase on stability / forced

stress testing)

Summarize all potential and actual impurities arising from the synthesis.

Identify impurities by Names, Structures, or RRT/HPLC.

Specify impurities as process impurities and/or degradants

Impurities: Acceptance Criteria

Release and Stability Specifications: Justification are based on

Pharmaceutical Development Studies Data

Analytical Data from Batches

Compendial Specifications (USP/EP Monographs)

Scientific Literature including EP, JP, etc

Safety Data on Qualification

Impurity specifications: Justification are based on

ICH Q3B and ANDA Guidance: Impurities in Drug Products

Level of Impurity observed in an FDA-Approved Drug Product (RLD),

batch analysis of the RLD close to expiry should be included

Significant Metabolite of Drug Substance

Scientific Literature including EP, JP, etc

Toxicology Studies

Compendial Specifications (Unspecified Impurities can not be

justified based on USP specifications)

Negotiation with FDA

Defining Impurities Safety: DS/ DP

ICH Q3A/B (R2) Threshold TerminologyListing, Reporting, Identification and Qualification of Degradation Products /

Drug Substances.

ThresholdThreshold for degradation products are expressed either

as % of drug substance or Total Daily Intake (TDI) of the

Drug Product.

Reporting

Threshold

A limit above (>) which a degradation product should be

reported.

Identification

Threshold

A limit above (>) which a degradation product should be

identified.

Achieving of the structural characterization or

qualitative parameter e.g. retention time in HPLC.

Qualification

Threshold

A limit above (>) which a Impurity/ degradation product

found to be toxicologically qualified.

Process of acquiring and evaluating data that

establishes the biological safety of an individual

degradation product or a given degradation profile.

See Next Decision Tree for Threshold Safety Studies

Yes

Yes

YesQualified

Structure elucidated?No

No

Related to others

with known toxicity?

Yes

No

No

Yes

No

Consider additional

testing or removal of

impurity

NoDecrease impurity

level below threshold

Yes

Toxicity documented

and sufficient?

Consider patient

population and duration

of use

Above threshold?

Adverse Effects?

Consider need for:

1. Genotoxicity studies (point mutation, chromosomal aberration)

2. General toxicity studies (one species, min 14 days, max 90 days

3. Other specific toxicity end point as appropriate

Qualified

Acceptable

Justification?

Qualified

Decision Tree

for

Threshold

Safety Studies

Start Here

Yes

Acceptance Criteria: Drug Substances

Drug Substances Acceptance Criteria : ICH Q3A (R2)Determine the Maximum Daily Dose (MDD)

Use MDD to calculate the ICH Thresholds for drug related impurities [ICH Q3A (R2)]

Maximum Daily

Dose (MDD) 1

Reporting

Threshold (RT) 2,3

Identification

Threshold (IT) 3

Qualification

Threshold (QT) 3,*

≤ 2g/day 0.05% 0.10% or 1mg per day

intake

(whichever is lower)

0.15% or 1mg per day intake

(whichever is lower)

≥ 2g/day 0.03% 0.05% 0.05%

1 The amount of drug substance, in the form of dose, administered per day.

2 Higher reporting thresholds should be scientifically justified.

3 Lower thresholds can be appropriate if the impurity is unusually toxic………….

That means Impurities below this level need not to be identified but Identification is

recommended, if it is acute toxic in nature.

* Impurities exceeding ICH Qualification Thresholds (QT)...... Try first if it is possible to

reduce the Level....If not possible, then Qualify – a process of evaluation of data which

establishes the Biological Safety of an individual impurity or an impurity profile..............

That means In all cases impurity At ICH Qualification Threshold must be QUALIFIED.

4 Ensure the Analytical Method is adequate to determine LOQ is equal or below RT.

5 Establish limits for “Individual Unspecified Impurity” to equal or below the IT.

6 Establish limits for each “Identified and/or Specified Impurity” to equal or below the QT.

Yes

No

Is impurity greater

than identification

threshold?

Yes

Structure

identified?

YesNo No

action

No

Greater

than qualification

threshold?

Any

known human

relevant risks?

No

Reduce to

safe level

Reduce

to not more than

() identification

threshold?

Reduce

to not more than

() qualification

threshold?

No further

action No action

Yes

No

Yes

No

YesIs the impurity

observed in an approved

drug substance and at a similar level

or is it adequately

qualified by other

approaches?

Consider patient population and duration of use and

consider conducting:

Genotoxicity studies (point mutation, chromosomal

aberration)a

General toxicity studies (one species, usually 14 to 90

days)b

Other specific toxicity endpoints, as appropriate

YesNoAny

clinically relevant

adverse effects?Qualified

Reduce to

safe level

No

Yes

No action

Decision Tree for Identification

and Qualification of Impurities in

Drug Substances (ICH Q3A(R2)

Quick Link for more details:

http://www.emea.europa.eu/pdfs/human/ich/273799en.pdf

ICH Q3A(R2)Start Here


Reporting Example 1 for 0.5 g Maximum Daily Dose

Reporting threshold = 0.05%, Identification threshold = 0.10%, and

Qualification threshold = 0.15%

Reporting Impurity Results for Identification and Qualification of Drug Substances

"Raw"

Result

(%)

Reported Result

(%) Reporting

Threshold

= 0.05%

Calculated Total

Daily Intake (TDI)

(mg) of impurity

(rounded result in

mg)

Action

Identification

(Threshold

0.10%

exceeded?)

Qualification

(Threshold

0.15%

exceeded?)

0.044 Not Reported 0.2 None None

0.0963 0.10 0.5 None None

0.12 0.12 + 0.6 Yes None +

0.1649 0.16 + 0.8 Yes Yes +

+ After identification, if the response factor is determined to differ significantly from the original

assumptions, it may be appropriate to re-measure the actual amount of the impurity present and

reevaluate against the qualification threshold (See Acceptance Criteria: Drug Substances Slide).

DS: Identification & Qualification/1



Qualification threshold = 1.0 mg TDI

Reporting Impurity Results for Identification and Qualification of Drug Substances

"Raw"

Result

(%)

Reported Result

(%) Reporting

Threshold

= 0.05%

Calculated Total

Daily Intake (TDI)

(mg) of impurity

(rounded result in mg)

Action

Identification

(Threshold 0.10%

exceeded?)

Qualification

(Threshold 1.0 mg

TDI exceeded?)

0.066 0.07 0.6 None None

0.124 0.12 1.0 Yes None +, *

0.143 0.14 1.1 Yes Yes +

+ See Identification & Qualification/1Slide

* To verify if a threshold is exceeded, a reported result has to be evaluated against the

thresholds as follows: when the threshold is described in %, the reported result rounded to the

same decimal place as the threshold should be compared directly to the threshold. When the

threshold is described in TDI, the reported result should be converted to TDI, rounded to the

same decimal place as the threshold and compared to the threshold. For example the amount of

impurity at 0.12% level corresponds to a TDI of 0.96 mg (absolute amount) which is then

rounded up to 1.0 mg; so the qualification threshold expressed in TDI (1.0 mg) is not exceeded.

DS: Identification & Qualification/2

Acceptance Criteria: Drug Products

Identify Impurity

Unspecified Impurity has limited acceptance

Determine (ICH Q3B – see next slide)

Maximum Daily Dose (MDD)

Reporting Threshold (RT)

Identification Threshold (IT)

Qualification Threshold (QT)

Ensure

Analytical Method is Adequate, LOQ is < Reporting Threshold

Establish

Limits for “Individual Unspecified Impurity” < Identification Threshold

Limits for “Specified Identified Impurity” < Qualification Threshold

Limits for “Specified Unidentified Impurity” < Qualification Threshold

Total Impurities

Qualify / Identify

Any impurity if a limit is greater than Qualification / Identification

Threshold

Degradation Product

(Degradation Impurity)

Specified Impurity

Unspecified Impurity

Unidentified

Impurity

Identified

Impurity

Yes

No

Is Degradation

Product greater

than identification

threshold?

Yes

Structure

identified?

YesNo No

action

No

Greater

than qualification

threshold?

Any

known human

relevant risks?

No

Reduce to

safe level

Reduce

to not more than

() identification

threshold?

Reduce

to not more than

() qualification

threshold?

No further

action No action

Yes

No

Yes

No

YesIs the degradation

observed in an approved

drug product and at a similar level or

is it adequately

qualified by other

approaches?

Consider patient population and duration of use and

consider conducting:

Genotoxicity studies (point mutation, chromosomal

aberration)a

General toxicity studies (one species, usually 14 to 90

days)b

Other specific toxicity endpoints, as appropriate

YesNoAny

clinically relevant

adverse effects?Qualified

Reduce to

safe level

No

Yes

No action

Decision Tree for Identification

and Qualification of Degradants

in Drug Product (ICH Q3B(R2)

Quick Link for more details:


ICH Q3B(R2)Start Here


Acceptance Criteria: Drug ProductsDrug Substances Acceptance Criteria : ICH Q3B (R2)

Determine the Maximum Daily Dose (MDD)

Use MDD to calculate the ICH Thresholds for drug related impurities [ICH Q3B (R2)]

Reporting Thresholds (RT)

Maximum Daily Dose (MDD) of API in Drug Product 1

≤1 g

> 1 g

Threshold Limit Based on TDI 2, 3

0.1% TDI

0.05% TDI

Identification Thresholds (IT)

Maximum Daily Dose of API in Drug Product 1

< 1 mg

1 mg – 10 mg

> 10 mg – 2 g

> 2 g


1.0% TDI or 5 μg, whichever is lower


0.2% TDI or 2 mg, whichever is lower

0.10% TDI

Qualification Thresholds (QT)

Maximum Daily Dose of API in Drug Product 1

< 10 mg

10 mg – 100 mg

> 100 mg – 2 g

> 2 g




0.2% TDI or 3 mg, whichever is lower

0.15%

1 The amount of drug substance, in the form of dose, administered per day.

2 Higher reporting thresholds should be scientifically justified.

3 Threshold for degradation products are expressed either as a percentage of drug substance or as Total Daily

Intake (TDI) of the degradation product. Lower thresholds can be appropriate if the impurity is unusually toxic.

Reporting Example 1 for 50 mg Maximum Daily Dose


Qualification threshold = 200 ug

Reporting Degradation Results for Identification and Qualification of Drug Products

"Raw"

Result

(%)

Reported Result

(%) Reporting

Threshold

= 0.1%

Total Daily Intake

(TDI) of the

Degradation

Product (rounded

result in ug)

Action

Identification

(Threshold

0.2%

exceeded?)

Qualification

(Threshold

200 ug TDI

exceeded?)

0.04 Not Reported 20 None None

0.2143 0.2 100 None None

0.349 0.3+ 150 Yes None +

0.550 0.6+ 300 Yes Yes +

+ After identification, if the response factor is determined to differ significantly from the original

assumptions, it may be appropriate to re-measure the actual amount of the degradation product present

and re-evaluate against the qualification threshold (See Acceptance Criteria: Drug Product Slide).

DP: Identification & Qualification/1

Quick Link for more details: http://www.emea.europa.eu/pdfs/human/ich/273899en.pdf



Reporting threshold = 0.05%, Identification threshold = 2 mg, and

Qualification threshold = 3 mg

Reporting Degradation Results for Identification and Qualification of Drug Products

"Raw"

Result (%)

Reported Result (%)

Reporting Threshold

= 0.05%

Total Daily Intake

(TDI) of Degradation

Product (rounded

result in mg)

Action

Identification

(Threshold 2 mg

TDI exceeded?)

Qualification

(Threshold 3 mg

TDI exceeded?)

0.049 Not Reported 1 None None

0.079 0.08 2 None None

0.183 0.18 + 3 Yes None +, *

0.192 0.19 + 4 Yes Yes +

+ See Identification & Qualification/1Slide

* To verify if a threshold is exceeded, a reported result has to be evaluated against the thresholds as

follows: when threshold is described in %, reported result rounded to the same decimal place as the

threshold should be compared directly to the threshold. When threshold is described in TDI, the reported

result should be converted to TDI, rounded to the same decimal place as threshold and compared to the

threshold . For Example an amount of 0.18% degradation level corresponds to a TDI of 3.4 mg impurity

(absolute amount) which is then rounded down to 3 mg; so the qualification threshold expressed in TDI (3

mg) is not exceeded.

DP: Identification & Qualification/2

Quick Link for more derails: http://www.emea.europa.eu/pdfs/human/ich/273899en.pdf



Example for Justification of Impurity Limits

Name Current

Lot

RLD Lot Proposed

Release

Limits

Proposed

Stability

Limits

Justification

Impurity A 0.80% 2.5% NMT 1.5% NMT 2.5% Metabolite

Impurity E 0.40% 1.0% NMT 1.0% NMT 1.0% Equivalent to

RLD

Any

Unknown

Impurity

< 0.07% < 0.05% NMT 0.20% NMT

0.20%

ICH Q3B

Identification

Threshold ++

Total

Impurities +

1.3% 3.6% NMT 2.5% NMT 3.5% Below RLD

+ Process-related impurities B, C, D, and F are excluded from the calculation of impurities

in the drug product.

++ The maximum daily dose of RLD is 64 mg/day and recommended Identification

Threshold is 0.2% or 2 mg TDI (Therapeutic Daily Intake/ Total Daily Intake) and

Qualification Threshold is 0.5% or 200μg TDI.


Testing should be conducted on various samples of marketed drug

product over the span of its shelf life (one sample near its expiration date)

Individual Unspecified Impurities in the drug product at release and for

stability should not exceed ICH Q3B “IT” based on MDD.

The acceptance criteria for Individual Specified Impurities for release

and stability should be the same as the drug substance unless the

impurities are shown to increase during manufacture or over time. Any

impurities that increase over time and are allowed > QT will need to be

qualified for safety. If the accelerated stability data fail tightened limits full

term long term data should be provided.

Example: MDD (Maximum Daily Dose) for the DP is 24 mgDrug Product QT = 0.5%

Impurity A: NMT 0.15% (drug substance)

NMT 0.8% (drug product release)

NMT 1.0% (shelf-life).

MinimizationVariability, Traceability and Uncertainty

Objectives of Measurement in Health Care Industries

1. To Ascertain:

The Quality of a Product and the Control of Process

2. To Demonstrate:

Potency of Product, Compliance with Specifications, and Conformity with

Regulations.

3. To Fulfill Requirements:

Pharmacopoeias and Guidelines

Goals of Determination of a Parameters

1. Testing: 2. Measurement

Test Results in Arbitrary Units Analytical Results in SI units

Method Dependent Method independent

Accepted Method Traceability

Reproducibility Uncertainty

Variability & MinimizationNo Matter What….Corporate must shows that

Analytical Instrument is Qualified

Computer System and Software is Validated

Audit Trail and Data Access is Limited

Data Storage (Electronic and Paper) and Traceability

Method is Validated & Purity-Stability Profile Indicator

SOP is Adequately Defined and Approved

Potency Assays for Generic Pharmaceutical

Method Feasibility

Method DOE and Development

Method Verification for Variables

Method Validation

Method Transfer for Re/Verification of Variables

Additional Validation / Revalidation

Monitor / Access Method Life Cycle

Quality Measurements via Technical Communication & Evaluation Evaluate potential uncertainty of quantitative and qualitative results and variables

Reduce variables as much as possible

Identified and optimize key variables using cause and effects diagram

Variability & Traceability in HPLC

P

R

O

B

L

E

M

Ruggedness

Complexicity of SOP / Protocols

Suitability of mobile phase

Accuracy of preparation of standards

Accuracy of preparation of samples

Matrix interference

R

E

S

U

L

T

S

Method Materials Environment

People Measurement LC Equipment

Stability and Purity

Storage and Handling

Sampling of test materials

Standard contamination

Sample contamination

Glassware contamination

Wrong glassware selection

Wrong reagents selection

Lab temperature

Lab humidity

Light exposure

Air borne contamination

Laboratory layout

Training

Failure to understand protocol

Work precision and accuracy

Individual interpretation of SOP

Lack of Communication

Poor housekeeping

Column degradation,

temperature fluctuations

UV detector lamp with

variable performance, air bubbles

Pump, flow rate, blockage, leakage,

trapped air, improper mixing,

Injector reproducibility

Time since certification

Time since sampling / dilution

Repeatability interference

Noisy baseline, carryovers

Density difference in solutions

Improper correction factor entry

Develop Yourself - Knowledge Based Methods and Tools from ICH Q9 Quality Risk Management

Potential Source of Method Variability: Ishikawa Diagnostic Methodology

»»»»»»»» R o o t C a u s e & E f f e c t F r a m e W o r k Analysis »»»»»»»»

Assay & Imp: Variability & Traceability

»»»»»»»» R o o t C a u s e & E f f e c t F r a m e W o r k Analysis »»»»»»»»

P

R

O

B

L

E

M

Diluent and Mobile Phase

Standard/ Sample Prep /

Resolution Prep/ Sensitivity Check

Runtime/ re-equilibration-X

Filtrate discard volume-X

Pipette Techniques-N

Weighing Techniques-N A

S

S

A

Y

&

I

M

P

Method Materials Environment

People Measurement Equipment

Reagents-source , grade, lot-N

Column and Vials-N

Volumetric glasswares-N

Transfer Pipettes-N

Filters & Syringe-N

Filtrate & glasswares-N

Weighing boat for Buffer-N

Product Type-N

Reference Material-C

Lab temperature-N

Lab humidity-N

Light exposure-N

Air borne contamination-N

Weighing Techniques -C

Integration Techniques-C

Equilibration Techniques-C

Variation in Extraction-C

Incorrect flask/Pipettes-C

HPLC Parameters-N

Sonication Time-N

Mechanical Shaker Action-N

Physical Balance Environment-N

Balance Accuracy-C

pH Meter Accuracy/Calibration-C

Run Time/ Equilibrium Time-X

Impurity calculation-C

Data Reporting-C

Suppress Integration Time-C

Noisy Baseline-C

Software Quality-C

Improper Correction Factor –C

Sampling Rate-X

N= Noise factor for Ruggedness, C= Control, eXperimental parameters for Robustness

Potential Source of Variability in Assay and Impurities Method

:

21 CFR 211.165 (a) 3 :

Federal CFR Guidance

Most Frequent referred Code of Federal Regulationshttp://www.gpoaccess.gov/cfr/index.html

http://www.betterchem.com/title21.htm

21 CFR 211.165

(e)

Analytical Method Validation

The accuracy, sensitivity, specificity and reproducibility of test method employed

by the firm shall be established and documented. Such validation and document may be accomplished in accordance with § 211.194 (a) (2).

21 CFR 211.166

(a) (3)

Stability Indicating Analytical Method

There shall be a written testing program designed to assess the stability

characteristics of drug products. The results of stability testing shall be used in

determining appropriate storage conditions and expiration dates

21 CFR 211.194

(a) (2)

Regulatory Guidance on Method Transfer

Methods used in the testing of the sample meet proper standards of accuracy

and reliability as applied to product tested.

The suitability of all testing methods used shall be verified under actual

conditions of use.

21 CFR 58.190

(a, b, c, d)

Regulatory Guidance on GLP

All protocol, raw data documentation, generated reports shall be retained,

archives, individual shall be indentified to archives. etc.

21 CFR 211.160

(a, b)

Laboratory Control Systems

Deviations from test procedure and written specifications.

http://www.gpoaccess.gov/cfr/index.html

http://www.betterchem.com/title21.htm

Quick International References

ICH Quick Link: Http://www.emea.europa.eu/htms/human/ich/ichquality.htm

Analytical Validation:

ICH Topic Q 2 (R1) Validation of Analytical Procedures: Text and

Methodology


Impurities:

ICH Topic Q 3 A (R2) Impurities in new Drug Substances


ICH Topic Q 3 B (R2) Impurities in New Drug Products


ICH Topic Q 3 C (R3) Impurities in Residual Solvents


Specifications:

ICH Topic Q 6 A Specifications: Test Procedures and Acceptance Criteria

for New Drug Substances and New Drug Products: Chemical Substances


ICH Q6A Decision Trees

http://www.ich.org/LOB/media/MEDIA431.pdf











http://www.ich.org/LOB/media/MEDIA431.pdf



Stability:

ICH Topic Q 1 A (R2) Stability Testing of new Drug Substances and

Products


ICH Topic Q1B Photostability Testing of New Active Substances and

Medicinal Products


ICH Topic Q1C Stability Testing: Requirements for New Dosage Forms


ICH Topic Q 1 D Bracketing and Matrixing designs for Stability Testing of

Drug Substances and Drug Products


ICH Topic Q 1 E Evaluation of Stability Data


ICH Topic Q 1 F Stability Data Package for Registration Applications

in Climatic Zones III and IV
















Regulatory Acceptance:

ICH Topic Q 4 B Regulatory Acceptance of Analytical Procedures and/or

Acceptance Criteria (RAAPAC)


ICH Topic Q 4 B Annex Annex to Regulatory Acceptance of Analytical

Procedures and/or Acceptance Criteria (RAAPAC)


Pharmaceutical Development:

ICH Topic Q 8 Pharmaceutical Development


ICH Topic Q 8 Annex Pharmaceutical Development


ICH Q9 document on Quality Risk Management

http://www.emea.europa.eu/Inspections/docs/ICHQ9Step4QRM.pdf

ICH Topic Q 10 Pharmaceutical Quality System















USA Guidelines Links

CDER Master Index Page

http://www.fda.gov/cder/guidance/index.htm

Analytical Procedures and Methods Validation Chemistry, Manufacturing,

and Controls Documentation

http://www.fda.gov/cder/guidance/2396dft.htm

Reviewer Guidance Validation of Chromatographic Methods

http://www.fda.gov/Cder/guidance/cmc3.pdf

Guidance for Industry, Bioanalytical Method Validation

http://www.fda.gov/cder/guidance/4252fnl.pdf

National Archives and Records Administration (CFR)


USP<1225> Validation of Compendial Procedures * Login Required


USP<1226> Verification of Compendial Procedures * Login Required















CTD and eCTD Informational Links

M4: Organization of the CTD

http://www.fda.gov/cder/guidance/4539O.PDF

eCTD Submissions

http://www.fda.gov/cder/guidance/7087rev.pdf

Drug Substance

http://www.fda.gov/cder/guidance/3969DFT.pdf

Drug Product

http://www.fda.gov/cder/guidance/1215dft.pdf

Q8: Pharmaceutical Developmentwww.fda.gov/cder/guidance/6746fnl.pdf

CTD-Efficacy

http://www.fda.gov/cder/guidance/4539E.pdf

CTD-Qualityhttp://www.fda.gov/cder/guidance/4539Q.PDF

QbR-Quality Overall Summary Outlinehttp://www.fda.gov/cder/ogd/












http://www.fda.gov/cder/guidance/4539Q.PDF

http://www.fda.gov/cder/ogd/

international analytical methodology

Technology

manufacturing process

process stream

lab way

product performance

verification methods

analytical activities

pat way

historical data