ANALYTICAL TEST METHODS

- A Workshop
 CONTENTS
• Introduction
• Analytical Method Development
• Principles of Method Transfer
• Stages of Technology Transfer
Process
• Few Examples
• Potential Problems and How to avoid
them
 Analytical Method
• What is an Analytical Method?

– It could be defined as collection of

instructions that completely describe
the sample analysis process such that
analyst can prepare the same type of
sample and produce equivalent results.
 Analytical Method
• Factors attributing to Error in analysis
– Incorrect weighing & transfer of analytes and
standards
– Inefficient extraction of analyte from matrix
– Incorrect use of pipettes, burettes or volumetric
flask for volume measurement
– Improper calibration of instruments
– Failure to use analytical blank
– Failure to remove interference by excipients in
the measurement of analyte
 Analytical Method
• What questions Analytical methods must
answer?
– Is the identity of drug in the formulated
product correct

– What is the % of stated content of drug product

present in formulation

– Does the formulation contains solely the active

ingredient or are additional impurities present
 Analytical……...Contd..
– What is the stability of a drug in the
formulation and hence the shelf life of the
product.

– Do the Identity & Purity of Drug substance or

excipient meet specifications

– What are the concentration of specified

impurities in the pure drug substance
Analytical Method Development

• GOAL
– Method and Specification should be
designed to adequately Evaluate desired
Quality Attributes

• CHALLENGE
– To develop Analytical Methodologies that
provide Accurate & Reliable data in Minimal
Time.
 Quality By Design
• Help us better describe the proposed product and
process design and its relation to the intended use
– improve process of establishing regulatory
specifications
• Improve our ability to identify and understand
critical product and process factors
– improve understanding and confidence in risk
mitigation strategies
• Allow us to utilize risk based approaches,
recognize good science and facilitate continuous
improvement
• Improve communication and systems thinking
– More efficient review and inspection process
 Desired State
 Product quality and performance achieved
and assured by design of effective and
efficient manufacturing processes
 Product Specifications based on mechanistic
understanding of how formulation and
process factors impact product performance
 An ability to effect Continuous Improvement
and Continuous "real time" assurance of
quality
Progression towards the ‘desired state’
is optional, and will be a continuum
 Analytical … Strategy
 Adoption of compendial Methods, if
applicable
or
 Development of alternative Method
or
 Adoption of Active drug substance
methodologies incorporating Special
dosage form Requirements
or
 Develop in House stability Indicating
method.
Analytical Method Development
General Considerations

1. To have clear criteria of method themselves,

in terms of
 Analysis time,
 Robustness and reliability,
 Sample preparation,
 Acceptable technology and
 SHE considerations.
Analytical Method Development
General Considerations

2. To have Understanding of Customer’s

current status and capabilities.

3. To ensure that the methods are suitable for

their intended purpose i.e. by validating the
method.
(Accuracy, Precision, Specificity, Detection
limit, Quantitation limit, Linearity and Range)
Analytical Method Development
Phase of Drug Development

 Initial Phase
 Ensure Potency ( relate directly to requirement
of known dose)
 Identify Impurities (relate to drugs safety
profile)
 Evaluate key drug characteristics such as crystal
form, drug release, drug uniformity ( relate to
drug Bioavailability)
Analytical Method Development
Phase of Development
 Later Phase
 Method should be stability indicating
 Capable of measuring the effect of key
manufacturing parameters to ensure consistency.

 Ultimately
 Method must be Robust, cost effectiveness,
transferable, and of sufficient accuracy and
precision.
Analytical Method Development
 Methods Transfer Protocol
 Method Transfer protocol should include
following:
 Objective, Scope, Responsibilities
 Materials, Methods, Equipment
 Experiment Design
 Acceptance criteria
 Documentation (report forms, what data,
outputs)
 Deviations
 References
 Signature / Approval Page
 Reference samples, Actives, Intermediates, &
Finished products
 Acceptance Criteria
Acceptance criteria must be established
prior to the method transfer

 Should be based on the

Intended use of the method,
Validation of the method
Historical data generated by the sending unit
 Methods Transfer - ASSAY
 Experimental Design – Acceptance Criteria

 At least two analysts (if available) at each

laboratory to independently analyze three lots (if
available), in triplicate (18 different executions of
the method).
 For products with multiple strengths, bracketing
may be appropriate (ICH Q1B, Q1D).
 Each analyst should use a different set of the same
instrumentation and/or columns (if available).
 Each analyst should independently prepare all
solutions.
 All applicable system suitability criteria should be
met.
 Acceptance criteria should be statistically derived
(should include comparison of mean and variability
of the results).
 Methods Transfer – Content Uniformity
 Experimental Design – Acceptance Criteria
 If the method for Content Uniformity is equivalent to assay
method, then a separate method transfer for assay is not
usually required.
 At least two analysts at each laboratory to independently
analyze at least one sample lot, in triplicate (6 different
executions of the method).
 For products with multiple strengths, bracketing may be
appropriate (ICH Q1B, Q1D).
 Each analyst should use a different set of the same
instrumentation and/or columns (if available).
 Each analyst should independently prepare all solutions.
 All applicable system suitability criteria should be met.
 Acceptance criteria should be statistically derived (data
variance & RSD)
 Impurities/Degradants/Res. Solvents
Experimental Design – Acceptance Criteria
 At least two analysts at each laboratory to independently
analyze at least three sample lots, in duplicate on different
days.\
 Each analyst should use a different set of the same
instrumentation and/or columns (if available).
 Each analyst should independently prepare all solutions.
 All applicable system suitability criteria should be met.
 LOQ should be confirmed by the receiving unit, in addition to
the response factors.
 Chromatograms from both laboratories should be compared to
ensure similar impurity profile (RRTs, peak response).
 Samples tested at both labs are similar in age, homogeneity,
packaging, storage.
 Acceptance criteria should be statistically derived (should
include comparison of mean and variability of the results).
 Method Transfer
Types of Method Transfer:

 Comparative Testing

 Covalidation between two laboratories

 Method Validation at receiving site

 Method Transfer
 Comparative Testing

– Most common form of method transfer

– Both the sites carry out testing as per pre

approved testing protocol using the same
sample and standard batches

– Data generated is compared against

predetermined acceptance criteria &

– Equivalence between two group is assessed.

 Method Transfer
 Covalidation Between Two Laboratories

– Receiving site is involved in the validation of

the methods being transferred
– e.g. in reproducibility studies (i.e. precision between
laboratory studies) used to validate the method.

– Receiving site, therefore is qualified to

routinely run the method without further
evaluation
 Method Transfer
 Method Validation at the receiving site

– Receiving site repeats the validation

experiments
– Following successful validation, the receiving
laboratory is considered qualified to perform
the method.
– Time consuming exercise
– used in microbiological methods
 Transfer Waiver
– Receiving Laboratory is already testing the
product and is thoroughly familiar with
procedures.

– Personnel who develop the method move from

one site to other site

– Method validation package encompasses the

new methods.

– New methods involve changes that do not

substantially alter the ability to use the method.
Stages of Technology Transfer
Process
• Stage 1 : Analytical Strategy

• Stage 2 : Knowledge and Information

Transfer

• Stage 3 : Transfer Protocol

• Stage 4 : Analytical Testing

• Stage 5 : Summary Report

 Flow Diagram
Formation of Method Discussion, Comment
Transfer Team and Review

Agree Analytical Transfer analysis

Transfer strategy

Knowledge & Information Summary Report

Transfer

Discussion, comment,
review, training Review and finalize

Method Transfer Transfer Approval

Protocol
 Stage 1
• Formation of Method Transfer Team
– Should include experienced individuals form critical
group
– Develop analytical transfer strategy
– provision of documentation for transfer process

• Analytical Transfer Strategy Document

– Originating site to prepare this document
– provide commitment to both time and resources to
support the transfer
– List of methods to be transferred and whether or not
they require formal transfer
 Stage 2
• Knowledge and Information Transfer
– Covers both training and documentation
– Summary report of how method is developed
– Full method details

• Method Discussion and Review

– Mainly be recipient site
– Enables recipient site to check the method and
suitability of equipment
– Training may be required in case of new and complex
methods

• Once transfer package is approved, it should be formally

approved by analytical transfer team
 Stage 3
• Method Transfer Protocol
– Give specific details of the testing to be performed,
samples to be utilised and acceptance criteria against
which the data will be assessed
– Appropriate Testing Design should be agreed upon
– Analytical samples for method transfer should be
identified and stored under appropriate condition
• Acceptance Criteria
– Key in demonstrating the equivalence of data
generated.
– Statistics can be used to set acceptance criteria
• Approval of Method Transfer Protocol
 Stage 4
• Analytical Testing
– Both the sites must perform the agreed testing on
the designated samples and generate data
– Checklist to ensure all the methods, documentation,
equipment, samples, standards and reagents are in
place
– Procedure for handling OOS results must be in
place.
– Changes if any to agreed protocol must be
investigated and documented
 Stage 5
• Summary Report
– Once transfer analysis is complete and all required data
have been generated , summary report must be written
– Normally written by receiving site.
– Summary of method qualifying the success or failure to
meet their acceptance criteria.
– Reviewed by both the sending and receiving site
• Transfer Approval
– Formal acceptance that the methods have been
successfully transferred to the receiving site
– and receiving site is now considered to be able to apply the
methods

ORGANISE THE CELEBRATIONS!!!!!!

 Examples of Technology Transfer
 Protocol for Dissolution Test for Tablets
1. Introduction
2 Purpose
• To describe the ruggedness evaluation for the transfer of
the Dissolution Tablets from sending unit personnel to
Receiving unit Personnel.
• To establish documented evidence that the results
generated by the personnel of receiving unit and sending
unit on identical samples are equivalent within
predetermined acceptance criteria.

3 Sponsors and Testing facility

• Sponsor - Testing lab
• Testing Facility
 Examples………. Contd...
4. Responsible Person
 Validation Director
 Responsible Analysts
 Quality Assurance
5. Materials And Equipment

6. Experimental Design
Method TestDosage Batch Number
2 mg EXM56098A
STM-AS-O Dissolution 2 mg EXP55098B
 Examples………. Contd...

7. Methodology
 Refer to method (STM-AS-O), perform one (1) Six(6)
unit dissolution for each batch.

8. Acceptance Criteria
 The absolute difference between the average of the
dissolution results at the Q value (20 minute) does not
differ more than 6.0% between two laboratory groups.

 Results should be reported to nearest whole number.

 Examples………. Contd...
9. Documentation and Archiving
 Both lab to summarize the results
 Sending unit will prepare a transfer report containing
following:
 Reference to the protocol
 Results tabulated for each lot and a summary table
 Statement including all qualification status.
 Approval signatures from both the sending and
receiving lab. (indicating agreement with the
qualification statements included in the document)
10. Amendments and Deviations
 No changes to be made unless approved by validation
director, change if any should be recorded
 Any deviations from protocol should be recorded and
reported.
 Examples………. Contd...
Dissolution

Batch EXM56098A (2mg) EXP55098B (2mg)

Vessel ABC XYZ ABC XYZ

1
2
3
4
5
6
Average %

Difference
Pass or Fail
 Assay :
System suitability – Precision
Injection No. Peak Area Injection No. Peak Area
1 _______ 6 _______
2 _______ 7 _______
3 _______ 8 ______
4 _______ 9 ______
5 ________ 10 _____ _
11 _______ _
12 _______
Mean Peak Area: __________ Mean Peak Area: __________
Standard Deviation:________ Standard Deviation:________
% RSD(First 5 Inj.):________ % RSD(All Inj.):___________

• Data file name

• Note book reference
• Analyst: ________________ Date: __________________
• Checked By_____________ Date: __________________
• Supervisor_______________ Date: __________________
 Assay :
System suitability – Resolution
Resolution is calculated using below equation:

Resolution = 1.18(t2 - t1) x 60

(w2 + w1)

t1 = Retention time of Drug Sub ( Lot 1) = _______min.

t2 = Retention time of Drug Sub ( Lot 2) = _______min.

w1 = Peak width at 1/2 height of Drug Sub ( Lot 1) = ______Sec.

w2 = Peak width at 1/2 height of Drug Sub ( Lot 1) = ______Sec.
Potential Problems and How to avoid them

1. A Good Quality Method

2. Language and Culture

3. Communication

4. Documentation & Procedure

5. Practical Problems

6. Acceptance Criteria
 1. A Good Quality Method
 Method must be well written

 Level of details is very important

 Nothing should be left for Interpretations

 Sample preparation procedure should be clear

and unambiguous

 More carefully the method is scrutinised, better

the chances of successful transfer
 2. Language and Culture

 For methods to be transferred to various countries

it is very important that face to face discussions are
understood by both the sides and
misunderstandings are avoided.

 People take note of cultural difference and adapt

them in discussions
 3. Communication
 Proper means of communication must be used
 Not to over rely on emails as means of
communication
 Some face to face meetings should be arranged,
videoconferencing can be desirable.
 It must be made clear who the correct contacts are
 Strong working relationship between the people
involved in the transfer activities from both sides.
 4. Documentation & Procedures
 Transfer process must be supported by efficient
document management

 Both the sites should use correct and same

versions.

 Biggest problem is that of difference between SOPs

used at the originating and receiving site.
 e.g. Statistical procedure can be different at both the sites
 treatment and handling of out of Specifications result
can be different
 5. Practical Problems
 Differences in fundamental equipment such as
filters, ferrules etc.
 HPLC Column performance can differ with
different manufacturers.
 Use and application of different data systems at
two sites
 e.g. Chromatography data system used to
integrate the chromatograms
 Difference in software packages e.g. Microsoft
word or Excel
 5. ……. Contd….
 Difference in Laboratory Information
management systems
 Difference in reporting formats, systems may
calculate the results in different ways.
 Major source of problem in method transfer
exercise in the inappropriate setting of acceptance
criteria
 Summary
 High Quality Method
 Good communication
 Involvement of both site as early as possible
 Clear process and agreed roles and responsibilities
 Well designed testing protocols with appropriate
acceptance criteria
 Concise and clear documentation
 Change control procedures
 FAQs
Q1 :The product assay specification approved in
the NDA is the average of 10 individual unit
falls between 90-110% of label claim.
• A firm reports the following low and high assay
values for a particular batch 87.3% and 101.6%
however average is 94.8%
– The firm tests 20 additional units and records
88.8% and 112.1%, average of 30 units is 95.3%.
– No further investigations and no change made
to label expiry date.
– Does the batch fail its assay specification? Is it
a concern?
 Answer
• Individual assay values that are outside the assay
specification do not constitute failure of test (Specification
is on average and not individual Value).

• But the matter should be investigated to determine whether

there was intent to provide less than 100% label claim ( if so
this is cGMP violation)

• Causes to be determined:
Batch records should be examined
In-Process test results should be checked
Operators to be interviewed
Reconciling inventory
sampling/testing variability from the beginning to
end of the lot
confirming adequacy of method
elevating any effect on expiration date
 Answer
– Company should have a written procedure to
deal with it

– All samples taken must be representative of lot

or batch.

– Appropriate sampling plans

– Batch history should be examined to see if it is a

recurring problem

– Process redesign and/or stricter in-process

control may be necessary.
 FAQs
Q2) Can it be assumed that USP chromatographic
assay methods are stability- indicating?
• No. While frequently assay methods utilizing
chromatography are stability-indicating, clearly
such is not always the case.
• USP acknowledges this in <1151> Pharmaceutical
Dosage Forms," which states, "Monograph
assays may be used for stability testing if they
are stability- Indicating..." Where use of a
stability- indicating assay method is required,
the user of the method must have data generated
from the testing of his/her product, showing that
the method distinguishes the active ingredient
from its degradation product(s).