Transcript International Site Video Conference

6th Annual Science and Standards Symposium
January 16, 2013
Istanbul
USP Reference Procedure
Sameer G. Navalgund, Ph.D.
Analytical R&D Laboratory
Topics

USP-MC Monographs

What is a Reference Procedure?

Critical characteristics of a Reference Procedure

Reference Procedure Development
– Process
– Solubility
• Dissolution of solid oral drug products
– Procedure Development
– Forced Degradation Studies
– Use of advanced technologies
2
The MC Monograph
 MC



monograph typically contains…
Tests, Procedures, and Acceptance Criteria
Tests are consistent with USP-NF (Identification, Assay, Impurities,
Performance Tests, Specific Tests)
Tests are grouped into one of three separate sections
• Performance Based Monograph (Core section)
• Reference Procedures (containing monograph specific procedures for
Assay and Impurities)
• Acceptable Procedures (containing procedures meeting requirements of a
criteria-based procedure, but not those of a reference procedure)

Procedures differ from USP-NF and include both
• Reference Procedures and
• Criteria-based Procedures

Acceptance Criteria are based upon the ICH Q6A, Q3A, and Q3B
limits.
Need of a Reference Procedure - I

Analytical procedures for pharmaceutical products and
components are typically derived from an understanding
of the manufacturing process, i.e., what’s likely to be
present.

These procedures are optimized to be as fast and simple
as possible.

Where a party other than the manufacturer wishes to test
a component or product, this type of detailed information
is not available.
4
Need of a Reference Procedure - II

To complicate matters further, the global pharmaceutical
marketplace includes components and products
processed from different starting materials, having
different manufacturing processes, and formulated using
different excipients and strengths.

This diversity leads to vast array of specific tests and
procedures to analysis nearly identical substances.

The goal of establishing a single reference procedure for
each test in a pharmacopeia has therefore been a difficult
one.
5
MC Approach – Reference Procedure

The MC approach deviates from tradition by
developing reference procedures from a different
perspective:
– Rather than adapting a procedure that has been
optimized to be fast and inexpensive by ignoring a
single manufacturer data;
– Our labs develop and implement a procedure that is
optimized to find all significant components in a
substance through separation and multidimensional
detection.
– A Reference procedure is developed using a battery of
analytical tools that are appropriate for the molecule
and matrix.
– HPLC / UHPLC + Photodiode Array (PDA) and Mass
Spectroscopic (MS) detection in series
– Evaporating Light Scattering Detector (ELSD), Corona
Detector
6
Critical Characteristics of Reference Procedure
Target article
For Development
Monograph
prepared using
General Chapter
<10>
Authentic
samples of the
article
Thorough
Literature
Review
Procured from at
least 2
independent
sources
Pharmacopeias
More sources are
better
Reported
synthetic and
degradation
routes
Published
literature
7
Monograph for Development - PBM
Monograph for
Development
PBM
Dynamic
Document
• Prepared by Scientific Liaison
• Tests for Identification, Assay, Impurities, etc.
• Acceptance Criteria, wherever applicable
• Possibilities for changes as we go forward
during method development or
• If additional information is available
8
“Proposed For Development” Monograph

List of “For
Development”
Monographs on
the MC website
9
A Typical “Proposed For Development” Monograph
Reference Procedure
10
Reference Procedure Development
Pre Lab
Literature Search
- Pharmacopeia–USP, EP, BP, JP
- Scientific Literature and Online
and Paper Journals
Procurement
- Drug substance, Drug products,
Impurities by Standards Acquisition
department
- Pharmacopeial impurities by
Analytical Development department
In Lab
- Method Development by
multi-source materials
- Specificity studies
- LC-MS Analysis for the
forced degradation studies
- Method finalization
- Method validation
- Data processing
Post Lab
Share the Method
validation data with the
Scientific Liaison
11
- Acid
- Alkali
-Thermal
- Peroxide
- Light
- Separation of
degradation
products
(Identification,
wherever possible)
- Loops back to
Method
development,
if needed
- Method
development
completed
- Transfer to
the Scientific
Liaison
- If MC monograph
is common to
USP-NF, it
provides additional
new impurities for
USP-NF
Method Validation and Data Processing
-LC-PDA-MS
-GC-MS
- Forced
degradation
studies
Method Finalization
- Solubility
- Spectroscopic
Identification
- Assay
- Related
Substances
- Use of multi
dimension
detectors /
orthogonal
detectors
Specificity Studies
Method Development
Reference Procedure Development – In Lab Details
Method Validation
- Linearity
- Accuracy
- Precision
-Day 1
-Day 2
-Day 3
Data Processing
-Review of
the
validation
data
-Transfer to
Scientific
Liaison
12
Method Development
Method Development
- Solubility
- Spectroscopic
Identification
- Assay
- Related
Substances
- Use of multi
dimension
detectors /
orthogonal
detectors
-LC-PDA-MS
-GC-MS

Solubility
– Different pH buffer solutions (1.2, 4.5, 6.8)

Spectroscopic Identification
– FT-IR, Specific Optical Rotation, etc.

Assay
– Chromatography

Related Substances
– Chromatography

Use of multi dimension detectors /
orthogonal detectors
– LC-PDA-MS, ELSD, Corona, etc.
13
Identification - Clarithromycin
14
Method Development
Method Development – Assay and Related Substances
- Solubility
- Spectroscopic
Identification
- Assay
- Related
Substances
- Use of multi
dimension
detectors /
orthogonal
detectors

Several criteria
– Quantification of important analytes
–
–
-LC-PDA-MS
-GC-MS
–
–
• Active substances, impurities, other
components
The most prevalent method is
chromatographic separation with
spectroscopic or spectrometric detection.
The procedure is adjusted to achieve
acceptable separation of the primary peak
from other peaks.
Use a multi-dimensional detector such as LCPDA with a full scan (200-400 nm or 200-700
nm)
Use a mass spectrometric compatible mobile
15
phase
Use of MS Compatible Mobile Phases
• Routine use of MS detector to evaluate a sample for
non-chromophoric species.
• Procedure development with mass spec-compatible
mobile phases.
• The resolving power of these mobile phases may be
less than phosphate buffers, but the information
obtained from the MS is considered more important that
the pure separation.
• When Liquid chromatography cannot be used or where
adequate separation cannot be accomplished with a
mass spec-compatible buffer, then it will be important to
consider likely interferences to a procedure and ensure
that the procedure can adequately quantify the analyte
of interest even in the presence of these interferences.
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Specificity Studies
Specificity (Forced Degradation) Studies
- Forced
degradation
studies

–
–
–
–
–
- Acid
- Alkali
-Thermal
- Peroxide
- Light
- Separation of
degradation
products
(Identification,
wherever possible)
- Loops back to
Method
development,
if needed
Forced degradation studies
Acid
Alkali
Thermal
Peroxide
Light (UV & Fluorescence)
• Solution
• Solid

Separation of degradation products
– Identification, wherever possible

Loops back to Method development, if
needed
17
Method Development, Forced Degradation Studies - I
Separation of Clarithromycin &
impurities
Acid degradation of Clarithromycin drug substance
18
Method Development, Forced Degradation Studies - II
Acid degradation of Clarithromycin drug substance
•Easier quantification and basic identification
information with full scan PDA and MS
•Detection of co-eluting species
19
Method Finalization
Final Method
- Method
development
completed
- Transfer to
the Scientific
Liaison
- If MC monograph
is common to
USP-NF, it
provides additional
new impurities for
USP-NF

Forced degradation completed
– Non-targeted screening
– Unequivocal separation

Method development completed

Internal Review

Method of analysis is transferred to
the Scientific Liaison

Types of Reference Standards
– Process impurities (Known / Unknown)
– Degradation products
20
Method Validation
General Chapter <10>
21
Method Validation – Assay - I
– Assay:
• Precision and Accuracy: six independent
weighings, two injections per sample
– Calculation and Acceptance Criteria: Calculate %
RSD and calculated predicted content of the Precision
measures using the calibration curve from the Range
requirement*
• Ruggedness: Precision and Accuracy
assessments over 3 day
• Range: six independent weighings at 80%, 90%,
100%, 110%, and 120%. The precision and
accuracy is determined for each concentration*
*the Precision and Accuracy is then compared to defined acceptance criteria of the Test to
determine the probability of the procedure returning a passing value if it was 100%. The
probability must be greater than 95%
22
Method Validation – Assay - II
• Specificity: Resolution of NLT 1.5 between main
and all impurities
– Process Impurities: evaluation of candidate material at
120% to 200% of the analytical concentration
identification (of presence) of impurity peaks at 0.01% or
greater
– Degradation Products: Target 30% reduction in active
content using Acid, Base, Heat, Light (UV and ambient),
Peroxide, others as appropriate: identification (of
presence) of impurity peaks at 0.10% or greater
– Un-retained and non-eluted Impurities: HPTLC with
attention to the Origin and Solvent front
• Ancillary Data
– FT-IR Spectrum, Water Determination , Enantiomeric
purity , Solubility in pH 1.2 and 6.8 buffers
23
Method Validation – Impurity Limit Procedure
• Precision: six independent weighings spiked at
the limit, two injections per sample
– Calculation and Acceptance Criteria: Calculate %
RSD: NMT an amount calculated in <10> (for a 0.1%
impurity, NMT 5.7%; for an 0.05% impurity, NMT 6%;
etc)
• Limit of Detection: five injections each of; one
sample spiked at the limit; and one sample spiked
at (limit - %RSD above)
• Acceptance criteria: the mean value of the
second standard must pass the limit test
• Specificity: each impurity must be quantifiable to
within the LOD
24
Method Validation – Quantitative Impurities Procedure - I
• Precision: six independent weighings spiked at
the limit, two injections per sample
– Calculation and Acceptance Criteria: Calculate %
RSD: NMT an amount calculated in <10> (for a 0.1%
impurity, NMT 2.8%; for an 0.05% impurity, NMT 3%;
etc)
• Ruggedness: Precision and Accuracy
assessments over 3 day
– Calculation and Acceptance Criteria: Calculate %
RSD: NMT an amount calculated in <10> (for a 0.1%
impurity, NMT 5.7%; for an 0.05% impurity, NMT 6%;
etc)
25
Method Validation – Quantitative Impurities Procedure - II
• Accuracy: six independent solutions spiked at
50%, 75%, 100%, 125% and 150% of the limit for
each specified impurity
– Calculation and Acceptance Criteria: Calculate Spike
Recovery and each concentration: between 100-%RSD
to 100+%RSD (for a 0.1% impurity; 97%-103%;this is
under consideration for a change to 94%-106%)
• Specificity: Resolution of NLT 1.5 between all
impurities (where unattainable, meeting Accuracy
is sufficient)
26
Data Processing and Transfer

Review of the
validation data

Transfer to the
Scientific Liaison
27