2013-09-18 usp stability 2 procedures

II. Developing Stability I di ti T t M th dIndicating Test Methods

Critical Stability Systems

Regulations ProceduresRegulations(I)

Procedures(II)

Stability Systems

Operations(III)

Investigations(IV)

Copyright © USP. All rights reserved.

Discussion Topics

2. Stability Indicating Test Procedures

Impact of analytical testing in the drug developmentImpact of analytical testing in the drug development process.

Review ICH Q2 A&B guidelines for methodReview ICH Q2 A&B guidelines for method development/validation.

Discuss Q3 for impurities monitoringDiscuss Q3 for impurities monitoring

How to develop stability-indicating test methods


Pharmacopoeia Monographs

• All compendial testing for API and drug product properties that are dependent on the manufacturing process should be validated or verified (e.g. process impurities, particle size, residual solvents, dissolution, p , p , , ,uniformity).

• For drug products, lack of interference from the placebo should be demonstrated. (Verification)

• If the procedure is implemented exactly according to th h lid ti i i d l llthe monograph, no validation is required legally.


Pharmacopoeia Monographs

•If the procedure is not stability-indicating, then an lt t t bilit i di ti d h ld balternate stability-indicating procedure should be

validated. • In the U S USP procedures are considered to be fully. In the U.S., USP procedures are considered to be fully validated; however, cGMP requires that individual laboratories demonstrate their ability to reliably execute the procedure.the procedure.


Definitions of Validation: USP

USP chapter <1225>:

“Validation of an analytical procedure is the process byValidation of an analytical procedure is the process by which it is established, by laboratory studies, that the performance characteristics of the procedure meet the requirements for the intended analytical applications.”


Definitions of Validation: ICH

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

“The objective of validation of an analytical procedure is to demonstrate that it is suitable for its intended purpose ”purpose.


Subpart I – Laboratory Controls

• Sec 211.160 General Requirements– Establishment (and change) of specs, standards,

sampling plans test procedures is requiredsampling plans, test procedures is required.– Must be followed and documented at time of

performance– Deviation must be recorded and justified.

• Sec 211 165 Testing and Release for distributionSec 211.165 Testing and Release for distribution– Conforms to specifications– Free of objectionable microorganisms– Sampling and testing plans are described in written

procedures– Must have acceptance criteria

8

Must have acceptance criteria– Validation must be done and written according to

211.194

Definitions of Validation: ICH

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

“The objective of validation of an analytical procedure is to demonstrate that it is suitable for its intended purpose.”

Guidance for Industry: Analytical Procedures and Methods Validation CMC Documentation:“M th d lid ti i th f“Methods validation is the process of

demonstrating that analytical procedures are suitable for their intended use”

9

A Recurring Theme for Validation

A basic theme for the definitions of validation indicates that validation pertains to demonstrating that the

th d imethod or process is

“…suitable for its intended use”


Critical First Step for Validation

It is essential that the purpose of the method to beIt is essential that the purpose of the method to be validated is clearly stated at the outset of the validation process.

As per ICH Q2(R1):

“The objective of the analytical procedure should be clearly understood since this will govern the validation characteristics which need to be evaluated ”characteristics which need to be evaluated.


Statement of Validation Conditions

It is also appropriate that the conditions are provided under which the validation was performed. If conditions are found that mitigate against using the method, they should be i di t dindicated.


Statement of Validation Conditions

From the section on robustness, ICH Q2(R1):

“If measurements are susceptible to variations in analytical conditions, the analytical conditions should be suitably controlled or a precautionary statement should be includedcontrolled or a precautionary statement should be included in the procedure. One consequence of the evaluation of robustness should be that a series of system suitability parameters (e g resolution test) is established to ensureparameters (e.g., resolution test) is established to ensure that the validity of the analytical procedure is maintained whenever used.”


Regulatory Requirements

Section 211.166(a)(3) requires that stability test

methods be reliable meaningful and specific whichmethods be reliable, meaningful, and specific, which

means that the content of active ingredient, degradation

products, and other components of interest in a drug

product can be accurately measured without

interference, often called "stability-indicating.“


ICH Guidelines

Q1AR2…”The testing should cover, as appropriate, the physical, chemical, biological and microbiological tt ib t ti t t d f ti lit t tattributes, preservative content, and functionality tests

(e.g for a dose delivery system). Analytical procedures should be fully validated and stability indicating.”


ICH Guidelines

Q2A… “Validated analytical procedures should be used, irrespective of whether they are for in-process, release, acceptance, or stability testing”.


ICH Guidelines

Q3B: “Analytical methods should be validated to demonstrate that impurities unique to the new drug substance do not interfere with or are separated from pspecified and unspecified degradation products in the drug product.”


Regulatory Requirements

FDA Draft Stability Guidance -- 1998

“…Validated quantitative analytical methods that can d t t th h ith ti i th h i l h i ldetect the changes with time in the chemical, physical, or microbiological properties of the drug substance and drug product, and that are specific so that the contents of active ingredients degradation products and otherof active ingredients, degradation products, and other components of interest can be accurately measured without interferences.”


Q2B: Methodology

Main objective is to demonstrate the procedure is suitable for its intended purpose.

Provides recommendations on how to consider the various validation characteristics for each of the 4various validation characteristics for each of the 4 analytical procedures specified in Q2A.– Identification test– Impurities (quantitative & limit)– Assay/Dissolution– Specific Tests– Specific Tests


Q2A: Analytical Procedures and Method Validation – August 2000 NON COMPENDIAL ANALYTICAL METHOD

Types of Procedures

ICH:

Finding and Recognizing the Tests

1. Identification tests2. Quantitative tests for impurities' content3 Limit tests for the control of impurities3. Limit tests for the control of impurities4. Quantitative tests of the active moiety in drug substance

or drug product or other selected component(s) in the drug product.

USP:Category 1: Like ICH 4Category 2: Like ICH 2,3Categor 3 Performance tests like dissol tionCategory 3: Performance tests like dissolutionCategory 4: Like ICH 1

Validation Characteristics

ICH:

The Characteristics are the Same

ICH:Accuracy Precision (repeatability

USP:Accuracy Precision (repeatability,

intermediate precision, and reproducibility )

yPrecisionSpecificity Detection LimitSpecificity

Detection Limit Quantitation Limit

Detection Limit Quantitation Limit Linearity RQuantitation Limit

Linearity Range

Range

Range

<1225> Validation of Compendial Procedures

Performance Characteristic

Category I Category II Category III Category IV

Quant Limit Test

Accuracy

Yes

Yes

*

*

No

Precision Yes Yes No Yes No

Specificity Yes Yes Yes * Yes

LOD No No Yes * No

LOQ No Yes No * No

Linearity Yes Yes No * No

Range Yes Yes * * No

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

Validation Characteristics

SPECIFICITYACCURACYLINEARITYLINEARITYRANGE

PRECISIONPRECISIONRepeatabilityIntermediate PrecisionPrecisionReproducibility

DETECTION LIMITDETECTION LIMITQUANTITATION LIMIT

ROBUSTNESS

24

ROBUSTNESS

Q1A(R) - Drug Substance --Stress Testing

• Stress testing is required– To understand the drug substance stability– To establish degradation pathways – To validate the stability indicating power of the

analytical procedures usedanalytical procedures used. – To support the severe conditions that may be

encountered during distribution.


Specificity

Q2A: “Stress studies (e.g. products of acid and base hydrolysis, thermal degradation, photolysis, oxidation) for the drug substance and for the active ingredient in g gthe drug product should be provided to demonstrate the specificity of the assay and analytical procedures for impurities. “


Specificity

Goals– Generate typical degradation products which may be

expected on stability at sufficient levels to allowexpected on stability at sufficient levels to allow identification

– Avoid secondary degradation– Target range is 5-20 % loss of active as judged byTarget range is 5 20 % loss of active as judged by

assay relative to an undegraded sample– Look for purity and mass balance


Typical Forced Degradation Design

Drug Substance• Solid State:

Drug Product:– Heat

– Heat– Photostability

Humidity

– Photostability– Humidity– Using placebo as control– Humidity

• Solution State: d di

Using placebo as control

• For combination product (multiple activedepending on

solubility– Acid

(multiple active ingredients)– Stress should be done for

API i di id ll d l– Base– Peroxide– Photostability

API individually and also in the presence of the other API (s).

28

Photostability

Evaluate Method Specificity with Stressed Samples

Are any of the degradants co-eluting/interferencing?

Determine the purity of the major analyte peakp y j y p

Direct Evaluation of peak purity (PDA vs MS)

Indirect Evaluation of peak purityIndirect Evaluation of peak purity

Spike in known degradation products (if available)


Example : System Suitability Sample

For impurity method of drug product with two API’s containing spiked standards

0.006 A)

B)

1.00

0.005 AP

I (A

AP

I (

0.200.400.60

0.80Full scale

Sensitivity check

0.003

0.004

1 A1

mp

A1

Dim

er(B

)0.0010.00 20.00

Critical Rs

ComponentAt 0.1% for DPS/N > 10

0 001

0.002 DG

B DG

Im

scomponent

0.000

0.001

2 00 4 00 6 00 8 00 10 00 12 00 14 00 16 00 18 00 20 002.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00

M.W. Dong, Modern HPLC for Practicing Scientists, Wiley, 2006, Chap. 9.

Case Study on Method Validation

An early-phase composite methodAn early-phase composite method0.035

86.

792

1.00Column: Supelco Ascentis C18

(3 100 4 6 i d )API

0.035

86.

792

1.00Column: Supelco Ascentis C18

(3 100 4 6 i d )API

0.025

0.030

6.39

8

AU

0.00

0.50

Minutes0.00 5.00 10.00 15.00 20.00

(3 μm, 100 x 4.6 mm i.d.)Mobile Ph.: 25 – 65% B in 15 min

A: 0.05% TFAB: 0.03% TFA in ACN

Flow Rate: 1.0 mL/min @ 30C

API

0.025

0.030

6.39

8

AU

0.00

0.50

Minutes0.00 5.00 10.00 15.00 20.00

(3 μm, 100 x 4.6 mm i.d.)Mobile Ph.: 25 – 65% B in 15 min

A: 0.05% TFAB: 0.03% TFA in ACN

Flow Rate: 1.0 mL/min @ 30C

API

AU 0.015

0.020

.170

1

Detection: UV at 230 nm Sample: 30μL of API at 1.0 mg/mL

Impurity 1

Precursor

AU 0.015

0.020

.170

1

Detection: UV at 230 nm Sample: 30μL of API at 1.0 mg/mL

Impurity 1

Precursor

0 005

0.010

127

590

8.8.

550

10.1

70

11.9

63

13.8

71

0 005

0.010

127

590

8.8.

550

10.1

70

11.9

63

13.8

71

0.000

0.005

5.1 7. 8 1

0.000

0.005

5.1 7. 8 1

M W Dong Modern HPLC for Practicing Scientists Wiley 2006 Chap 8


-0.005

Minutes2.00 4.00 6.00 8.00 10.00 12.00 14.00

-0.005

Minutes2.00 4.00 6.00 8.00 10.00 12.00 14.00

M.W. Dong, Modern HPLC for Practicing Scientists, Wiley, 2006, Chap. 8.

Accuracy (Recovery)

Accuracy: at least 9 determinations over at least 3 concentrations covering the specified range (e.g., 3 concentrations x 3 replicates)p )Linearity/Range: a minimum of five concentrationsDetermine across a range that at least encompasses the specification of the testthe specification of the testAcceptance criteria must be realistic.


Accuracy (Recovery) Highlights

• Accuracy (recovery) & Linearity• Placebo method (placebo is spiked with 3

concentrations of 3 sample weights each of API)• Analytes can be adsorbed (excipients, colorants, manuf.

Process) on surfaces and filters => analyte recoveriesProcess) on surfaces and filters => analyte recoveries• Errors can also be caused by equipment, reagents,

standards, analysts, etc.standards, analysts, etc.

33

Linearity highlights

• May want to study high level and low level linearity separately. (Beer’s law)

• Low level linearity• Low level linearity– If the reporting limit is to be 0.05% and the

specification limit is to be 0.10% (i.e. API), if linear i d 0 05% t 0 12%is done 0.05% to 0.12%

– Not practical• Correction factors for impuritiesp

Precision

• Repeatability:– System Precision y– Method Precision

• Intermediate Precision: different days, analysts, equipment.– Evaluate individually and compare both

• Reproducibility• Reproducibility• Ruggedness• RobustnessRobustness

• Acceptance criteria – Must be realistic

35

p

Intermediate Precision

Degree of reproducibility of test results as a function of the variables

F (x) = f(A) * f(D) * f(E) * f(C)

Intermediate Precision Evaluation for

Bulk Substance Assay by HPLC1.6

0.4

0.8

1.2

% R

SD

0

Analy

st

Day

Equip

ment

Column


Eq

Robustness of HPLC Methods

• “… susceptible to variations in analytical conditions…”

• Sample Preparation Robustness

• Chromatographic System Robustnessg p y

• Report analyte levels and show chromatograms

• optimized during development parameters are• optimized during development, parameters are interactive

• Could use an experimental design for this study• Could use an experimental design for this study.


Sample Preparation Robustness

Examine different sample preparation schemes

– solvents shake vortex sonication timesolvents, shake , vortex sonication time, grinding/homogenizing, weights & volumes

Solution stability – time, temperature, light, solvent vs. y , p , g ,fresh solution

– Measure both potency and impurity levelsp y p y

– Needed for standard and drug product sample solutions


Chromatographic Robustness

Evaluate chromatography vs. nominal conditions– Columns (lots, suppliers), mobile phase pH, ionic

strength, % organic, gradient profile, flow rate, column, injector, detector temperature, injection volume, split flow, wavelength, Instrument brands,volume, split flow, wavelength, Instrument brands, etc.

Some method parameters can be changed +/- from nominal conditions

Can matrix test parameters, i.e., change several method parameters simultaneouslymethod parameters simultaneously


Potential Impurities

Impurities for APIs and Excipients: • synthesis precursorsy p• synthesis bi-products• residual solvents• catalysts• catalysts• decomposition• and other impurities

Impurities for Drug Products: • degradations productsg p• extractables• residual solvents• unknown substances


• unknown substances

Q3 – Impurities

Q3A (R): Impurities in New Drug Substances (2/11/03)Q3B (R): Impurities in New Drug Products (11/14/03)Q3C: Impurities: Residual Solvents (12/24/97)Q3C: Impurities: Residual Solvents (12/24/97)

Tables and Lists (11/13/03)ICH Q3: Impurity LOQ must be less than Reporting

API DRUG PRODUCT

Threshold

≤ 2g/day > 2g/day ≤ 1g/day > 1g/day

Reporting Threshold 0.05% 0.03% 0.10% 0.05%Reporting Threshold 0.05% 0.03% 0.10% 0.05%

Identification Threshold 0.10% 0.05% 1.00% 1.00%

Qualification Threshold 0.15% 0.05% 1.00% 1.00%

Establish System Suitability Criteria

System Precision: minimum n=5 standard injection at the beginning of the run and precision throughout the run (b k ti i j ti )(bracketing injections)

Standard Check

Chromatographic Precision:

Should be more than one criteria

contain resolution of a critical pair (R ≥ 1.5)

Establish Tailing factor (< 2.0)

Approximate retention time

Need baseline separation of close peaks


Need baseline separation of close peaks

.

Establish System Suitability Criteria

Should be base on the known capabilities of the procedure

Should be developed with the average analyst in mind and for a routine laboratory operations.

Impurity method should include an assessment of LOQ of the least sensitive analyte.


Specificity (Impurities Tests)

There are two basic approaches to establishing specificity for procedures designed to measure impurities:impurities:

With impurity or degradation product standardsWithout impurity or degradation product standards


Specificity for Impurity Method

When impurities are available– Spiking drug substance or drug product with

appropriate levels of impurities– Demonstration of the separation of these impurities

individually and from other components in theindividually and from other components in the sample matrix


Specificity for Impurity Method

When impurities are not available– Demonstrate by comparing the test results of

samples containing impurities or degradation products to a second well-characterized procedure (pharmacopeial method, validated independent(pharmacopeial method, validated independent procedure)

– Include samples stored under relevant stress diti li ht h t h idit id/b h d l iconditions: light, heat, humidity, acid/base hydrolysis,

and oxidation


Specificity (Case Study -With Standards)

Drug XX Sample

EnantiomerEnantiomer Standard

47

Specificity (Case Study -Without Standards)

Drug XX Stressed sample

48

Detection Limit – Impurities

Q2(R1):

The detection limit (DL) may be expressed as: DL = 3.3 S/b;

Where S = standard deviation of the response obtained fromWhere S standard deviation of the response obtained from- Blank - Calibration curve (standard deviation of y-intercepts

of regression lines)- Calibration curve (residual standard deviation of a

regression line )ˆ( 2∑ yyg

f2

)(

−

−=∑

n

yyS i

ii

xy

and b = slope of the calibration curve

Quantitation Limit–Impurities

The quantitation limit can be determined using the same approaches as for the detection limit. For instrumental approaches Q2(R1) provides:approaches, Q2(R1) provides:

QL = 10 S / bQL = 10 S / b

Where S and b are similar to those defined before for DLWhere S and b are similar to those defined before for DL


Life Cycle of Analytical Methods

Change ofChange of Products Re-Validation

Change of Analytical

Validationy

Methods


Re-Validation Focus

Possible Causes of Analytical Method Change that warrant method revalidation

Synthetic process changeFormulation changeOut of Specification investigationOut-of-Specification investigationSpecificity problem (e.g., new impurity)Ruggedness issueggPost approval changes


Re-Validation Focus

Change control system: Levels of changes should be defined in SOP:

Level 1 : no action requiredLevel 2 : verification requiredqLevel 3 : revalidation required


Verification Concepts

Validation: Challenges the analytical method using a well defined sampleg

Verification: Challenges the analytical environment using a well defined method

– Analyst– InstrumentInstrument – Reagents– Matrix


General Requirements for Verification

From GC <1226>:

“Although complete revalidation of a compendial method is not required to verify the suitability of the method under actual conditions of use, some of the analytical performance characteristics listed in chapter <1225>, Table 2, may be used for the verification process.”, y p

“Only those characteristics that are considered to beOnly those characteristics that are considered to be appropriate for the verification of the particular method need to be evaluated.”


Robustness

Definition - The ability to remain unaffected by small but deliberate variations in method parametersparameters

Ruggedness vs. RobustnessThey are different

Ruggedness: Performance under normal variability

Robustness: small but deliberate changes


Key Factors for Change Control for Method Validation

Clearly define major and minor changes

Parameters to be revalidated depending on level of change

Equivalency of materials or instrumentation

Establish System Suit with robustness dataEstablish System Suit with robustness data

Document all changes and validation assessment efforts in Method Development report. Review previous changes of p p p gthe method before making modifications.

Understand your product’s chemistry.

Know what affects your product’s stability

For R&D, ensure that all methods are fully validated in their


, yfinal form prior to testing submission batches

2013-09-18 usp stability 2 procedures

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