Waters Executive UPLC Forum

17-18 May 2012

Hyderabad India
USP General Chapters Update
Hyderabad, India
USP General Chapters Update
Ravi Ravichandran, Ph.D.
Principal Scientific Liaison Small Molecules Principal Scientific Liaison, Small Molecules
1
Topics
Proposed Revisions to Chromatography <621>
Stimuli article in PF35(6) Nov Dec 2009 Stimuli article in PF35(6) Nov-Dec-2009
<621> revision proposal in PF 37(3) MayJ une 2011
<621> revision proposal in PF 38(2) Mar-Apr 2012
Harmonization of Chromatography<621> with EP & J P
Validation Verification Method Transfer Chapters
Definitions
Equivalent or Better concept
Transfer of Analytical Procedures
Relationship between the three chapters Relationship between the three chapters
2
Main points
Chromatography <621> describes in detail the range of
adjustments allowed in the system when the suitability
test failed
What if the prescribed column is no longer available
id ti b bt i d ith th or a more rapid separation can be obtained with another
column. Both these situations currently require
revalidation
Ideal scenario allows the flexibility to change column
dimensions or particle size as long as equivalent or
b tt l f i i t i d better column performance is maintained
USP published a Stimuli article in PF35(6)Nov-Dec
2009
3
2009.
Stimuli article PF 35(6)
Transfer of HPLC Procedures to Suitable Columns of
Reduced Dimensions and Particle Sizes
Uwe D. Neue, Doug McCabe, Vijaya Ramesh, Horacio Pappa, J im
DeMuth
ABSTRACT This Stimuli article contains proposals to help
the analyst adjust HPLC column length and particle size to
achieve separation power at least equivalent to that used in
the original procedure, markedly increasing the range of
options currently allowed in Chromatography <621>. The options currently allowed in Chromatography 621 . The
article presents the scientific rationale for application of these
proposals to isocratic procedures and follows with gradient
procedures
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procedures.
Examples
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PF 37(3) May-J un-2011 proposal
Based on stimuli article Transfer of HPLC Procedures to Suitable
Columns of Reduced Dimensions and Particle Sizespublished in Columns of Reduced Dimensions and Particle Sizes published in
PF 35(6) [Nov.Dec. 2009], pages 16221626.
A more flexible approach to select HPLC column dimensions
allowing the analyst to choose a combination of column length and allowing the analyst to choose a combination of column length and
particle size to achieve separation power equivalent to that obtained
using the prescribed column.
Approach increases the range of options currently allowed in the
chapter.
Helps reducing solvent consumption p g p
Reduces time of the analysis,
6
PF 37(3) May-J un-2011
Particle Size (HPLC): The particle size and/or the length of the column
may be modified provided that the ratio of length (L) to particle size (dp)
remains constant or varies not more than 25% with respect to the ratio
obtained with the column prescribed in the monograph. When particle
size is not mentioned in the monograph, the ratio must be calculated
using the largest particle size consigned in the USP definition of the using the largest particle size consigned in the USP definition of the
column.
Flow Rate: When the particle size is changed the flowrate may Flow Rate: When the particle size is changed, the flow rate may
require adjustment:
F
2
= F
1
(d
c2
2
d
p1
) / (d
c1
2
d
p2
)
Where F
1
and F
2
are the flow rates for the original and modified
conditions, respectively; d
c1
and d
c2
are the respective column
diameters, and d
p1
and d
p2
are the particle sizes.
7
PF 37(3) May-J un 2011
2
1
2
2
1 2
d d
dp dc
F F

=
Relative Values
2
2
1
dp dc
L, mm dc, mm dp, m L/dp F N Pressure Time
250 4.6 10 25,000 0.5 0.8 0.2 3.3
150 4.6 5 30,000 1.0 1.0 1.0 1.0
150 2.1 5 30,000 0.2 1.0 1.0 1.0
100 4 6 3 5 28 600 1 4 1 0 1 9 0 5 100 4.6 3.5 28,600 1.4 1.0 1.9 0.5
100 2.1 3.5 28,600 0.3 1.0 1.9 0.5
75 4.6 2.5 30,000 2.0 1.0 4.0 0.3
8
75 2.1 2.5 30,000 0.4 1.0 4.0 0.3
50 4.6 1.7 29,400 2.9 1.0 8.5 0.1
PF 38(2) Mar-Apr 2012
The proposal for particle size allowances was amended
based on comments received based on comments received
Additional changes are added:
Allowing the use of a guard column even when it is
not prescribed in the individual monograph.
Under System suitability, it is indicated what type of
adjustments are allowed in gradient conditions adjustments are allowed in gradient conditions.
Allowances to modify the particle size in liquid
chromatography are being modify.
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PF 38(2) Particle size changes
F i ti ti th ti l i d/ th For isocratic separations, the particle size and/or the
length of the column may be modified provided that the
ratio of the column length (L) to the particle size (dp) g ( ) p ( p)
remains constant or into the range between -25% to
+50% of the prescribed L/dp ratio. Alternatively, other
combination of L and dp can be used provided that the combination of L and dp can be used provided that the
number of theoretical plates (N) is within -25% to +50%,
relative to the prescribed column.
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PF 38(2) Mar-Apr 2012 : Use of Guard column
In LC procedures, a guard column may be used with the
following requirements, unless otherwise is indicated in
th i di id l h the individual monograph:
(a) the length of the guard column must be NMT 15%
of the length of the analytical column, and
(b) the packing material should be the same as the (b) the packing material should be the same as the
analytical column (e.g., silica) and contain the same
bonded phase (e.g., C18).
In any case, all system suitability requirements specified
in the official procedure must be met with the guard
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column installed
PF 38(2)Mar-Apr 22012: Isocratic vs gradient
Adjustments to chromatographic conditions: Isocratic vs j g p
gradient
Adjustments to the composition of the mobile phase in Adjustments to the composition of the mobile phase in
gradient elution may cause changes in selectivity and
should be made with caution. If adjustments are
h i l ki ( i t i i th necessary, change in column packing (maintaining the
same chemistry), the duration of an initial isocratic hold
(when prescribed), and/or dwell volume adjustments are ( p ), j
allowed. Additional allowances for gradient adjustment
are noted in the text below.
12
PF 38(2)Mar-Apr2012: Isocratic vs gradient
Adjustment Isocratic Gradient
pH of Mobile Phase Allowed Allowed
Concentration of Salts in Buffer Allowed Allowed
Ratio of Components in Mobile
Phase
Allowed Not allowed
Column dimensions (length,
diameter)
Allowed Not allowed
Particle size Allowed Not allowed
Flow rate Allowed Not allowed
Injection volume Allowed Not allowed
Column temperature Allowed Not allowed
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Column temperature Allowed Not allowed
Additional Committee activities on <621>
Stimuli article to be published in PF 38(3) May-
June 2012: Signal-to-Noise Measurements from
Chromatographic Data by J ohn V. Hinshaw and J ohn
W. Dolan
The article discuss the correlation between manual
and electronic (root-mean-square, RMS)
measurements measurements
Currently in USP:
S/N = 2H/h
14
Additional Committee activities on <621>
Stimuli article published in PF 38(3) [May-June 2012]:
Signal-to-Noise Measurements from Chromatographic Data
Recommendations:
Definition of S/N: S/N = S/N
p-p
f f The signal should be measured from the best estimate of the
center of the noise band to the highest point on the peak.
the noise measurement should be taken over an interval of 5
half-height peak widths or 30 s, whichever is larger.
RMS noise should not be used as the primary definition of S/N
in monographs in monographs
The use a conversion factor to convert p-p noise
measurements to RMS ones or vice versa, is discouraged as
th lt t li ti
15
the results are not realistic.
Additional Committee activities on <621>
Harmonization activities under the PDG initiated
EP is the leading pharmacopeia for this chapter
At this time pharmacopeias are evaluating Stage 3 draft
Some sections are easy to harmonize, others not so easy
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Comparison USP and EP
Parameter USP EP
Requirement after Chromsystem
adj stment
Verification
necessar
Meeting SS
req irement is adjustment necessary requirement is
adequate
Particle Size L/dp = constant
(proposed)
50% reduction
(proposed)
Internal diameter Change allowed as
long as linear
velocity is kept
+25%
y p
constant
Injection volume Decrease or
increase
Only decrease
17
(proposed)
Topics
Proposed Revisions to Chromatography <621>
Stimuli article in PF35(6) Nov Dec 2009 Stimuli article in PF35(6) Nov-Dec-2009
<621> revision proposal in PF 37(3) MayJ une 2011
<621> revision proposal in PF 38(2) Mar-Apr 2012
Harmonization of Chromatography<621> with EP & J P
Validation Verification Method Transfer Chapters
Definitions
Equivalent or Better concept
Transfer of Analytical Procedures
Relationship between the three chapters Relationship between the three chapters
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Validation - Verification - Transfer
<1225> Validation of Compendial Procedures
Validation will be required when
an analytical procedure is used to test a non-official article.
an official article is tested using an alternative procedure (see
USP General Notices 6.30).
<1226> Verification of Compendial Procedures
Verification will be required the first time an official article is tested
using a USP procedure.
<1224> Transfer of Analytical Procedures y
Transfer will applies when a non-compendial procedure is moved
from one lab to another.
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Definitions for Validation & Verification
<1225> Validation of Compendial Procedures: p
users of analytical methods described in the USP-NF are
not required to validate accuracy and reliability of these
methods, but merely verify their suitability under actual methods, but merely verify their suitability under actual
conditions of use. [21 CFR 211.194(a)(2)]
<1226> V ifi ti f C di l P d <1226> Verification of Compendial Procedures:
Verification consists of assessing selected analytical
performance characteristics, such as those that are
d ib d i h t 1225 t t i t described in chapter <1225>, to generate appropriate,
relevant data rather than repeating the validation process.
20
Basic concepts
Validation: Challenges the analytical method using a well
defined sample defined sample
Verification: Challenges the analytical environment using a
well defined method well defined method
Analyst (education, training, experience)
I t t Instrument
Reagents
Matrix
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Analytical Performance Characteristics
Accuracy
Precision Precision
Repeatability
Intermediate precision
Reproducibility p y
Specificity
LOD/ LOQ LOD / LOQ
Linearity
Range Range
Robustness
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Verification of Compendial Procedures <1226>
Applies to drug substances, drug products, and
excipients excipients.
Application: titrations, chromatographic procedures,
spectroscopic tests etc spectroscopic tests, etc.
Verification is not required for basic compendial test
procedures that are routinely performed unless there procedures that are routinely performed unless there
is an indication that the compendial procedure is not
appropriate for the article under test.
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Verification
Typical industry practices for verification of
compendial procedures
Specificity
Peak purity
Matrix
Accuracy: Recovery in the specification range
Precision: Repeatability in the specification range
LOD: Verification of detection at 50% of the specification
LOQ: Verification of quantitation at 50% of the specification
Usually not needed: Linearity, Range, and Robustness
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Risk-Based Approach
Verification requirements depend on the assessment of Verification requirements depend on the assessment of
Complexity of the procedure
Complexity of the material
Degree and extent of the verification process depends on
level of training and experience of the user
the type of procedure
associated equipment or instrumentation,
specific procedural steps specific procedural steps
article(s) are being tested
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Allowance for alternative procedures in USP
USP 31 - NF 26 General Notices:
C li b d t i d l b th f lt ti d Compliance may be determined also by the use of alternative procedures
choosen for advantages in accuracy, sensitivity, precision, selectivity, or
adaptability to automation or computerized data reduction, or in other
special circumstances. Such alternative or automated procedure shall be spec a c cu s a ces Suc a e a e o au o a ed p ocedu e s a be
validated.
USP 32 - NF 27 General Notices: USP 32 - NF 27 General Notices:
Alternative methods and/or procedures may be used if they provide
advantages in terms of accuracy, sensitivity, precision, selectivity, or
adaptability to automation or computerized data reduction, or in other adaptability to automation or computerized data reduction, or in other
special circumstances. Such alternative procedures and methods shall
be validated as described in the general chapter Validation of
Compendial Procedures <1225 > and must be shown to give
equivalent or better results
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equivalent or better results.
Equivalent or Better Concept
Option Name Demonstrating Comparison
to official
Number of
characteristics to official
procedure
characteristics
considered
1 Acceptable Procedures Acceptable No Many
2 Performance Equivalence
Equivalent or
Better
Yes Many
3 Results Equivalence Equivalent
Yes
One
4 Decisionequivalence Equivalent
Yes
One 4 Decision equivalence Equivalent One
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Reproduced from Acceptable, equivalent or better approaches for alternatives
to official compendial procedures by W. Hauck et al. PF35(3), 2009
Transfer of Analytical Procedures <1224>
Definition from the chapter:
The transfer of analytical procedures (TAP), is the
documented process that qualifies a laboratory (the
receiving unit) to use an analytical test procedure that
originated in another laboratory (the transferring unit), thus originated in another laboratory (the transferring unit), thus
ensuring that the receiving unit has the procedural
knowledge and ability to performthe analytical procedure knowledge and ability to perform the analytical procedure
as intended.
28
When transfer is needed?
R&D to QC labs
Site A to Site B (same company or not)
QC to Contract lab
29
Transfers options
I. Comparative testing
II. Co-validation
III. Revalidation
IV. Transfer waiver
30
Comparative testing
Comparative testing the common method
Performed with validated procedures
No challenge to validation parameters No challenge to validation parameters
Requires analysis of a predetermined number of
l f th l t b b th th t f i d samples of the same lot by both the transferring and
receiving units.
31
Covalidation
Transferring and Receiving units participate in
interlaboratory covalidation. y
Used when the procedure is not yet fully validated
G l h t V lid ti f C di l P d General chapter Validation of Compendial Procedures
<1225> provides guidance about which characteristics
are appropriate for testing.
Statistical evaluation of the results may be challenging
32
Revalidation
The receiving lab repeats some or all of the validation
experiments to challenge the applicable validation experiments to challenge the applicable validation
performance characteristics
More time consuming and may be more difficult to observe
differences between different sites, operators, and
instruments.
33
Transfer Waiver
New product comparable to an existing product
Receiving unit has experience in the test procedures
Analytical procedure transferred is the USP-NF
Verification should apply in this case (see <1226>). Verification should apply in this case (see 1226 ).
Analytical procedure transferred same or similar to a
procedure already in use procedure already in use.
The personnel in charge of the development, validation
or routine analysis of the product at the sending unit
moved to the receiving unit.
34
Procedure Transfer
Transfer
YES
Validated?
YES
Can transfer
be waived?
Transfer
waiver
(Option IV)
NO NO
be waived?
(Option IV)
Comparative
testing
(Option I)
Is transferring
site available?
Can
comparative testing
to be done?
YES
NO
R lid ti
Co validation
YES
35
Revalidation
(Option III)
Co-validation
(Option II)
Relationship between Validation / Verification/Transfer
Implementation of a
new procedure
Validated? <1225>
NO
Validated? <1225>
YES
Compendial? <1224>
NO
YES
36
<1226>
Looking into the future
USP t bli h d E t P l t t USP established an Expert Panel to generate
proposals for the revision of <1224>, <1225> and
<1226>
New chapters under development:
Statistical tools for validation Statistical tools for validation
Critical Validation Parameters for Acceptable Procedures
Acceptable Alternative Procedures
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Contact information
Dr. Horacio Pappa
Principal Scientific Liaison, General Chapters
[email protected]
38
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