Method Validation in the Drug development Process-김현성

8/2/2019 Method Validation in the Drug development Process-

1/54

Analytical Method Validation in The Drug

Development Process

Kim HyunSung Ph.D.Berna Biotech Korea

QC Chemistry lab


2/54

Contents

Introduction The importance of Analytical method in drug development

Drug development and GMP

Requirements of analytical method in drug development

Validation

What is validation? Analytical method validation

Analytical performance characteristics

Definitions

How to calculate characteristics?

Summary of validation protocol & report

When do you perform revalidation?

Conclusion


3/54

3

Pharmaceutical?

A pharmaceutical product manufactured by biotechnologymethods (involving live organisms; bioprocessing)

Introduction


4/54

4

Scope of Pharmaceuticals

Introduction


5/54

5

The Importance of Analytical Methodology in

the Drug Development Process

During the drug development, the evaluation of1) Determining optimum crystalline and salt form of the drug

substance, testing support to process development and

cleaning validation

2) The dosage forms bioavailability

3) Determining optimum formulation

4) The setting of specifications for the drug substance,

intermediates, and drug product

5) Identification, quantitation, and qualification of impurities and

degradants

6) The shelf-life of the product (stability)

7) Product safety

8) Support to preclinical and clinical studies (safety and efficacy)


6/54

6

The Importance of Analytical Methods in the

Drug Development Process

The chemistry, manufacturing, and controls section of a US

regulatory filing is required to contain the following directly

related to analytical testing:

1) Method validation

2) Physical description and characterization of the drug

substance and drug product-proof of the chemical structure

3) In-process controls

4) Characterization of reference standards

5) Specifications-description of analytical methods

6) Evaluation of container/closure system for storage7) Justification of drug product development

8) References to compendial test methods for inactive ingredients

9) Bioequivalence

10) Pharmaceutical development report


7/54

7

The Importance of Analytical Methodology in the Drug

Development Process

Physicochemical Characterization (1)

1) UV/visible or mass spectrometry

2) Amino acid analysis

3) Amino acid or Nucleic acid sequencing

4) Carbohydrate analysis and, if appropriate, sequencing

5) Peptide mapping

6) Determination of disulfide linkage

7) SDS-PAGE

8) Isoelectric focusing (1D or 2D)

9) HPLC, GC, LC, or thin layer chromatography

10) Nuclear magnetic resonance spectroscopy


8/54

8


Development Process

Physicochemical Characterization (2)

11) Assay to detect related proteins including deamidated, oxidated,

processed, and aggregated forms

12) other variants, such as amino acid substitutions and

adducts/derivatives

13) other process contaminants such as sulfhydryl reagetns, urea,

residual host proteins, residual DNA, and endotoxin

3 Stability indicating method

.


9/54

9


Development Process

Biological Characterization

1) Specific identity testing such as Western blot analysis or ELISA

2) Cytometric analysis

3) Neutrovirulence testing, if appropriate

4) Serotyping

5) Electrophretic typing

6) Inactivation studies : Irreversibility

7) Neutralization assays

8) Titration

9) in-vivo assay

10) in-vitro assay (ELISA)

11) Specific toxicity

12) Abnormal toxicity


10/54

10

The Importance of Analytical Methodology in


After the registration of drug product to regulatory authority, the

test methods are used in the pharmaceutical quality control

laboratory to do the following.

1) Identify the drug substance and product

2) Quantitate the pharmaceutical active ingredient

3) Determine level of purity

4) Guarantee the overall quality of the product


11/54

11

Q 1 : Quality ?

Ans.) 1) Lot (, )

2)

Why?

lot , ,

.

Question & Answer


12/54

12

Q 2 : Quality ?

Ans.)

1) (Safety)

2) (Stability)3) (Efficacy, Potency, Strength)

Q 3 : Quality ?

Ans.)

1)

2)

3)

Question & Answer


13/54

13

Stakeholders in Quality Analytical Testing

During Drug Development

Analytical

test

Processdevelopment

/Characteriza

tion/Controls

Support to

Clinical

Trials

Product

development

Toxicology

Pharmacokinet

ics

Regulatory

Filings

Stability /

Release


14/54

Drug Product Development and GMP

Full characterization

Full GMP

21 CFR 210, 211

21 CFR 610

Phase III

Phase I

Phase II

Pre-clinical

QA &QC, Clinical Monitoring Program

Validated MethodPre-qualified

method

(Standard screening

methods)


15/54

15

Q 4 : ?

Ans.) 2

Why? 2

.

Question & Answer


16/54

16

Q 5 : 2 fix

?

Ans.)

.

Question & Answer


17/54

17

Q 6 : Process development Product qualityupgrade ?

Ans.) lot comparability test ()

1) :

2) : ,

Q 7 : Process development Product qualityupgrade ?

Q 8 : ?

Question & Answer


18/54

18

Requirements of Analytical methods During


Depending on the stage of development, these analytical methods arestandard screening methods at the start of the development

process, which over time are gradually upgraded to thoroughly

validated methods for NDA application.

The filed methods must be simple, robust, and reliable- that is, easy to

use and perform without deviations when appropriately applied in aqualified laboratory.

Note

1. Screening methods are typically not optimized for speed and robustness.

2. The filed methods referred to as VTR2AP methods

(validated, transferable, robust, rapid, accurate, and precise)

James M. Miller and Jonathan B. Crowther, Analytical Chemistry in a GMP

envirinment: A practical guide, John Wiely & Sons,New York (2000)


19/54

19

Laboratory Analytical Methods Flow During


MethodValidation

Method

Transfer

MethodDevelopment

Approved


20/54

20

Q 9 : ?

Ans.)

. (R&D or QC)

r R&D

QC .

Question & Answer


21/54

21

Whats validation?

The FDA defines the term as :

Established documented evidence which provides a high degree of

assurance that a specific process will consistently produce a product

meeting its pre-determined specifications and quality attributes.

General Principles of Validation (1987)

ICH guideline defines the term as :

A documented program that provides a high degree of assurance

that a specific process, method, or system will consistently produce

a result meeting pre-determined acceptance criteria.

Q7A-GMP for active phamarceutical ingredients (2000)


22/54


23/54

23

Q 10 : Quality Specification ?

Ans.) Specification Quality .

Specification Product Quality .

Product Quality .

Question & Answer


24/54

24

Acceptance criteria

Acceptance criteria means

The product specifications and acceptance / rejection criteria,

such as acceptable quality and unacceptable quality level, with

an associated sampling plan, that are necessary for making a

decision to accept or reject a lot or batch (or any other

convenient subgroups of manufactured units)

CFR Part210.3 Definitions (2000)

ICH guideline define the term as :

Numerical limits, ranges, or other suitable measures for acceptance

of test results.

Q7A-GMP for active phamarceutical ingredients (2000)


25/54

Analytical Method Validation

Initial Method Validation Guidance Issued in 1987

Guideline for submitting samples and analytical data for methods

validation. Food and Drug Administration, February 1987. US

Government Printing Office:1990-281-794:20818.

Updated in August 2000 (Draft Guidance!)

Analytical Procedures and Method Validation. Fed. Reg. 65(169), 52,776-

52,777, 30 August 2000

CFR Part 211.165 Testing and release for distribution (e)

The accuracy, sensitivity, specificity, and reproducibility of test

methods employed by the firm shall be established and documented.

ICH guideline Q7

GMP guide for active pharmaceutical ingredients

ICH guideline Q2(R1)

Validation of analytical Procedure : Text and Methodology


26/54

ICH & USP Method Validation

Analytical Performance Characteristics

Validation parameter ICH USP

Accuracy O O

Precision

Repeatibility O O

Interm. precision O -

Reproducibility O -

Specificity of Selectivity O O

Detection limit O O

Quantitation limit O O

Linearity O O

Range O O

Robustness O O

Ruggedness - O


27/54

Definitions

Specificity:The ability to access unequivocally the analyte in the presence of

components which may be expected to be present. Typically these

might include impurities, degradants, matrix, etc.

Linearity:

Its ability to obtain test results which are directly proportional to the

concentration (mount) of analyte in sample within given range.

Range:The Interval between the upper and lower concentration (amounts) of

analyte in the sample ( including these concentrations) for which ithas been demonstrated that the analytical procedure has a suitable

level of precision, accuracy and linearity.


28/54

28

Definitions

Precision:

The closeness of agreement (degree of scatter) between a series ofmeasurements obtained from multiple sampling of the same homogeneous

sample under the prescribed conditions.

Accuracy:

The closeness of agreement between the value which is accepted either as a

conventional true value or an accepted reference value and the value found.


29/54

29

Definitions


30/54

30

Definitions

Limit of detection:

The lowest amount of analyte in a sample which can be detected but

necessarily quantitated as an exact value.

Limit of quantitation:

The lowest amount of analyte in a sample which can be quantitatively

determined with suitable precision and accuracy.Ruggedness:

The degree of reproducibility obtained under a variety of conditions, such as

different laboratory, different analysts, different instruments, environmental

conditions, operators and materials. (USP)

Robustness:

A measure of its capacity to remain unaffected by small but deliverate

variations in method parameters and provides an indication of its reliability

during normal use. (ICH)


31/54

Comparison of ICH & USP

Analytical Performance Characteristics

Characteristic Refer. ManEquip.

or Cond.Material

Inter-

lab

Intra-

lab.

Precision ICH same same same - O

Interm.Precision

ICH different different same - O

Reproducibility ICH different different different O -

Robustness ICH same

same /

different different - O

Ruggedness USP different different different O -


32/54

1. How to measure the Specificity?

Method : Impurity challenge and Recovery test Impurity : Hydrolyte of HBG by hydrogen peroxide

Challenge : half volume of hydrolyte

Calculation

Measured con. of challenge

Recovery yield = ------------------------------------------- X 100

Calculated con. of challenge


33/54

2. How to measure the Linearity?

The correlation coefficient(r2 or r), slope of regression line and

residual sum of squares should be submitted.

A minimum of5 concentrations is recommended.

Used the standards (5 con.) and samples (4 con.)


34/54

3. How to set the Range?

The specified range is normally derived from linearity studiesand depends on the intended application.

Acceptable degree of linearity, accuracy and precision.

The following minimum specified ranges should be

considered.

Forassay of active substance or a finished product,80~120% of concentration.


35/54

4. How to calculate the Accuracy?

Accuracy should be assessed on samples spiked with knownamounts.

Minimum 9 determinations over a minimum 3 concentration

level covering the specified range(e.g. 3 concentrations/3

replicate)

Should be reported as percent recovery by the assay of

known added amount of analyte in the sample or as thedifference between the mean and the accepted true value

together with the confidence intervals.


36/54

36

Q 11 : accuracy Spike test

?

Ans.) accuracy

.

.

280nm :

Lowry

. Bradford Lowry

.

Question & Answer


37/54

37

Q 12 : accuracy specificity Spike test

?

Ans.) challenge .

accuracy challenge

specificity challenge.

Question & Answer


38/54

5. How to calculate the Precision?

Repeatability Minimum 9 determinations covering the specified range(e.g.

3 concentrations/3 replicate) or

A minimum of6 determinations at 100% of the test

concentration.

Intermediate precision Depends on the circumstance under which the procedure is

intended to be used.

Establish the effects of random events on the precision.

Typical deviations to be studied include individually days,

analyst, equipment.


39/54

5. How to calculate the Precision?

Reproducibility

Assessed by means of an inter-laboratory trial.

Should be considered in case of standardization of an

analytical procedure.

This data is not part of the marketing authorization dossier

Recommended data

Standard deviation, relative standard deviation (coefficient ofvariation) and confidence interval should be reported for each

type of precision investigated.

t-test analysis


40/54

6. Detection limit

Based on visual evaluationMostly for non-instrumental methods

Based on signal-to-noise

Analytical procedures which exhibit baseline noise

Compare measured signals from samples with known low

concentration of analyte with those of blank samples.

A sinal-to-noise ratio 3 or 2:1 is acceptable.

Based on the Standard Deviation of the Response

and the Slope

The quantitation limit (QL) may be expressed as:

DL = 3.3/S

where = the standard deviation of blankS = the slope of the calibration curve

The slope S may be estimated from the calibration curve of

the standard.


41/54

7. Quantitation limit

Based on visual evaluationThe quantitation limit is generally determined by the

analysis of samples with known concentrations of analyteand by establishing the minimum level at which theanalyte can be quantified with acceptable accuracy andprecision.

Based on signal-to-noiseA typical signal-to-noise ratio is 10:1.

Based on the Standard Deviation of the Responseand the SlopeThe quantitation limit (QL) may be expressed as:

QL =10/S

where = the standard deviation of blankS = the slope of the calibration curve

The slope S may be estimated from the calibration curve ofthe standard.


42/54

8. How to measure the Robustness?

The evaluation of robustness should be considered duringthe development phase and depends on the type of

procedure under study. It should show the reliability of an

analysis with respect to deliberate variations in method

parameters.

Lowry : reaction time (30min vs. 40 min)

Measurement : confidential variance, standard

deviation, T-test


43/54

Summary of Method Validation Protocol

Parameter Sample Testing No SpecificationAcceptance

Criteria

AccuracyLow, Medium, High &

spiked L, M, H 6 Spiked recovery 80~120%

Repeatability 20, 50, 70, 90ug/mL 6 CV% a

Linearity5 Std &

20, 50, 70, 90ug/mL6

Correlation

coefficient>0.99

SpecificityHBG bulk treated with

H2O25 Recovery 80~120%

Range Low, Medium, High 5 Accuracy, Precision &Linearity pass

LOQ Blank 6 =(10*SD)/slope Calculated value

Robustness Medium 6 (2 person) t-test P> a


44/54

44

Q 13 : Protocol parameter LOD ?

Ans.) :

.

.

:

parameter .

Question & Answer


45/54

45

Q 14 : ?

Ans.) , (SD)

2SD, 3SD % .

200ug/mL, SD 13.3ug/mL 2SD

.

Acceptance criteria = [200 (2*13.3)] / 200 *100 = 86.7% (LL)

= [200 + (2*13.3)] / 200 *100 = 113.3% (UL)

* 3SD : Bioassay

Question & Answer


46/54

Summary of Method Validation Report

Parameter SpecificationAcceptance

CriteriaResult

Accuracy Spiked recovery 80~120% ave. 96.02%

Repeatability CV% a(0.05) L 0.768,M 0.293, H 0.293

LinearityCorrelation

coefficient>0.99 >0.995

Specificity Recovery 80~120% ave. 106.18%

RangeAccuracy, Precision &

Linearity pass 25~110ug/ml

LOQ =(10*SD)/slope Calculated value 2.11ug/ml

Robustness t-test P> a 0.652 > 0.05


47/54

47

Q 15 : validation protocol

?

Ans.)

1) (, , ) review

2)

3) :

4)

- acceptance

criteria ( .)

Why? worst case

acceptance criteria worst case

acceptance criteria .

Question & Answer


48/54

48

Q 16 : ?

Ans.) : System suitability test

1) Negative control (Blank)

2) Positive control (Standard material)

3) Reference (Normal product or Clinical product)

4) Calibration curve : linearity regression coefficient (r or r2)

Question & Answer


49/54

49

Q 17 : When do you perform revalidation?

Ans.)

Changes in the synthesis of the drug substance

Changes in the composition of the finished product Changes in the analytical procedure

ICHQ2(R1) Validation of analytical procedures: Text and

Methodology (2005)

In other case, some companies perform the method validation regular

interval.

Question & Answer


50/54

50

FY 99QS/GMP Deficiencies of Domestic Inspections(483s Issued)

20

22

23

23

25

29

31

36

36

40

42

44

0 5 10 15 20 25 30 35 40 45

Doc.Control

Processing Proc

Design/Dev Plans

Nonconform

Audits

Dev.Spec.

DHRCor/Pre Action

Design Changes

Validation

Inspection/Test Proc.

Complaints

Est. FY99 Total QS/GMP EIs with 483s issued=656

DeficiencyArea

Sample Size = 293


51/54

51

Q 18 : 483 ?

Ans.) Warning letter inspection GMP

Question & Answer


52/54

52

Q 18 : 483 ? ()

Question & Answer

The inspection revealed:

1. Analytical results were reported to [redacted] stating that a sample met

specifications when either out-of-specifications (OOS) results were

obtained on the sample analysis or on the quality control samples used

to determine the validity of the analytical results. These OOS results were

not investigated/documented properly to assure results reported to

[redacted] were accurate and valid. [21 CFR 211.165(a)]

2. Inadequate method validation in that OOS findings were discarded

without investigating the cause of the OOS results and analytical data

was selectively reported to support the validation. [21 CFR 211.165(e)]

3. Use of reference standards and reagent solutions for extended periods

of time without data in the analytical records supporting time of use. [21

CFR 211.194(c)]


53/54

53

Q 19 : 21 CFR 211.165(e)?

Question & Answer

e) The accuracy, sensitivity, specificity, and reproducibility of

test methods employed by the firm shall be established and

documented. Such validation and documentation may be

accomplished in accordance with CFR 211.194(a)(2)


54/54

Conclusion

GMP is Good Manufacturing Practice.

GMP is based on documentation.

Time is money.

Sometimes, Time is more valuable than money

in especially drug development process.

Documents

Method Validation in the Drug development Process-김현성