Module 06: Validation

Instructor Name

USP Affiliation: Position,

Committee
Title: Professional Title
Company: Company Name
Education: Degrees
Instructor Photo Degrees continued
Instructor bio…Natet pe volorib usapidus etur ad estruption porro ventorem
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eos sit aut am vid quaes re dolum unt voluptatem fugia dolup. ad estruption porro
ventorem doluptibusam dempore con etur, esecear uptatus eum rem volupta
doluptatur? Ectet eos sit aut am vid quaes re dolum unt voluptatem fugia dolup.
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Instructor bio continued if more space needed.. Natet pe volorib usapidus etur ad estruption porro ventorem
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unt voluptatem fugia dolup. ad estruption porro ventorem doluptibusam dempore con etur, esecear uptatus eum rem
volupta doluptatur? Ectet eos sit aut am vid quaes re dolum unt voluptatem fugia dolup.
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volupta doluptatur? Ectet eos sit aut am vid quaes re dolum unt voluptatem fugia dolup. ad estruption porro ventorem
doluptibusam dempore con etur, esecear uptatus eum rem volupta doluptatur? Ectet eos sit aut am vid quaes re dolum
unt voluptatem fugia dolup.
2
© 2018 USP
Disclaimer

Because USP text and publications may have legal implications in the U.S. and elsewhere, their language must
stand on its own. The USP shall not provide an official ex post facto interpretation to one party, thereby placing
other parties without that interpretation at a possible disadvantage. The requirements shall be uniformly and
equally available to all parties.
In addition, USP shall not provide an official opinion as to whether a particular article does or does not comply
with compendial requirements, except as part of an established USP verification or other conformity
assessment program that is conducted separately from and independent of USP's standard-setting activities.
Certain commercial equipment, instruments or materials may be identified in this presentation to specify
adequately the experimental procedure. Such identification does not imply approval, endorsement, or
certification by USP of a particular brand or product, nor does it imply that the equipment, instrument or material
is necessarily the best available for the purpose or that any other brand or product was judged to be
unsatisfactory or inadequate.
This course material is USP Property. Duplication or distribution without USP’s written permission is prohibited.
USP has tried to ensure the proper use and attribution of outside material included in these slides. If,
inadvertently, an error or omission has occurred, please bring it to our attention. We will in good faith correct
any error or omission that is brought to our attention. You may email us at: legal@[Link].
3
© 2018 USP
Learning Objectives – Module 6

 Validation concept of combined steps

– The dissolution step
– Sampling and filtration
– Sample transfer
– Analytical finish

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Outline

 Validation
– Specificity
– Linearity and range
– Accuracy/ recovery
– Precision
• Repeatability
• Intermediate precision
• Reproducibility
– Robustness
– Stability of standard and sample solution
– Considerations for automation
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Validation – Definition

 ICH Q2 (R1): “... The objective of validation of an analytical procedure is to

demonstrate that it is suitable for its intended purpose”
 Analytical Procedure: “…The analytical procedure refers to the way of
performing the analysis”
 “…It should describe in detail the steps necessary to perform each analytical
test ...” (e.g., sample, reference standard, reagent preparation, use of
apparatus, generation of the calibration curve, calculations)

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Validation – The Dissolution Testing in Detail

 Definition of validation needed for entire process

and sub-processes
– The dissolution part
– Drug substance and dissolution media
– Sampling, transfer, processing
– Chemical analysis
– Computation
 Validation of dissolution testing process requires risk analysis of all technical
steps
– From introduction of specimen - to computation of results

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Validation of Dissolution Procedures
Dissolution method – using a well-characterized dosage form
 Precision
 Robustness
Analytical method – using standard solution or spiked placebo
 Specificity - placebo interference
 Linearity and range
 Accuracy/Recovery
 Precision
 Robustness
 Solution stability
For combination products the dissolution procedure has to be validated for each
active ingredient
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Validation – ICH Q2 (R1) Guidance
ASSAY

- DISSOLUTION
TYPE OF ANALYTICAL (MEASUREMENT
PROCEDURE
ONLY)

- not required - CONTENT/POTENCY

+ required Characteristics
(1) if reproducibility was Accuracy
+
performed, intermediate
Precision
precision is not needed
Repeatability +
Interm. Precision
+ (1)
Specificity
+
Detection Limit
-
Quantitation Limit
-
Linearity
+
Range
+ 9
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Specificity/Placebo Interference

 Placebo constituents
– Spiked placebo in dissolution medium at 37C
– Interference doesn’t exceed 2%
 Dissolution medium
– Interference doesn’t exceed 1%
 Other active drug substances
 Significant levels of degradants

For extended release products a placebo version of the finished dosage form may
be used
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Specificity/Placebo Interference

Example UV analysis:

 Compare to the standard at 100% of the label claim with that of the placebo, and
a typical sample solution
 Calculate the percent interference
 The interference does not exceed 2%

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Specificity/Placebo Interference

 If the placebo interference exceeds 2%

– Alternative wavelength
– Alternative assay
– Subtraction of reading at reference wavelength
– Use of scanning spectrophotometry with second derivative
– Baseline subtraction (correction factor)
– Inactivation of the interference

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HPLC Specificity

 Chromatograms over an extended run time using the blank, placebo, standard
and sample solutions to identify late eluting compounds
 Representative chromatogram of dissolution medium, filtered placebo solution,
standard solution, and filtered dissolution sample solution
 Absence of interfering peaks in the placebo chromatogram

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HPLC Specificity

Blank solution – dissolution medium

Standard solution

Sample solution

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Linearity and Range

 Five standard solutions of the drug, ranging in concentration from -20% of the
lowest expected concentration for the dissolution profile to +20% of the highest
concentration during release from the dosage unit strength
 Solutions may be prepared
– Standard solution
– Spiked placebo
– By method of standard addition
 Not more than 5% (v/v) of organic solvent to enhance drug solubility for the
preparation of the standard solutions
 The highest concentration should not exceed the linearity limits of the equipment

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Linearity and Range

 Appropriate least-squares regression program

 To be reported
– Correlation coefficient
– Slope of the regression line
– y-intercept
– Residual sum of squares
– Trend analysis of residuals
 The square of correlation coefficient ≥ 0.98
∑𝓃𝓃 �
𝒾𝒾=1 𝓍𝓍𝒾𝒾 + 𝓍𝓍̅ 𝓎𝓎𝒾𝒾 + 𝓎𝓎
𝑟𝑟 = 𝑟𝑟𝓍𝓍𝓍𝓍 =
∑𝓃𝓃
𝒾𝒾=1 𝓍𝓍 𝒾𝒾 + 𝓍𝓍̅ 2 ∑𝓃𝓃 = 1 𝓎𝓎 + 𝓎𝓎
𝒾𝒾 𝒾𝒾 � 2

 The y-intercept must not be significantly different from zero at the 95%
confidence limit
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Linearity and Range - Example

Concentration level [%] Concentration Mean Absorbance

dissolution of dosage drug substance absorbance individual values
strength [µg/mL] (n=2)
0.7
40% 3.2315 0.12680 0.1286 0.6
0.1250
0.5
3.4924 0.13705 0.1378 0.4
0.1363

Abs
0.3
60% 5.0262 0.20000 0.2022 0.2
0.1978
0.1
4.9064 0.19335 0.1949 0.0
0.1918
-0.1 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16
80% 9.0030 0.35655 0.3600 Concentration µg/ml
0.3531

8.8412 0.35230 0.3561

0.3485 Slope: 0.03934 Abs/[µg/mL]
100% 13.2927 0.52825 0.5350
0.5215 Intercept: 0.00129 Abs
12.9117 0.50985 0.5139
0.5058 Cor. coef. r: 0.99992
120% 15.5259 0.61095 0.6180
0.6039 r2: 0.99984
15.6553 0.61265 0.6175
0.6078

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Accuracy/Recovery

 Samples containing the drug substance and any other constituents present in
the dosage form
– Individual solution prepared directly with dissolution medium
– Stock solution of drug substance in organic solvent and subsequent dilution with dissolution
medium
– Method of standard addition (spiked drug product)
 Range from -20% of the lowest expected concentration to +20% of the highest
concentration during dissolution
 Minimum of 3 concentration levels, each level tested in triplicate (n=3)
 The amount of placebo blend is to be the same as in the dosage form

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Accuracy/Recovery

 App. 1 and 2 - test the mixture of excipients and drug powder according to the
conditions specified in the method
 Poorly soluble drugs - stock solution - drug dissolved in organic solvent (max
5% final volume), and diluting with dissolution medium
 Stock solution transferred to the vessel
 Recovery is typically 95% to 105% of the weighed amounts
 Acid stage for Delayed-release dosage forms

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Accuracy - Example

% Theory %
 Paddle at 100rpm, HPLC Recovery
97.50 Average 97.25
 Dissolution medium heated to 37ºC 16.38 96.34 Lower 95% CL 95.22
97.92 Upper 95% CL 99.29
 Standard stock solution 81.88
97.94
98.08
Average
Lower 95% CL
97.86
97.21
97.57 Upper 95% CL 98.52
 1000 mL minus “x ml”dissolution 98.78 Average 98.31
medium given into a vessel, one 106.45 98.24 Lower 95% CL 97.20
97.90 Upper 95% CL 99.41
placebo tablet added 98.55 Average 97.58
131.80 96.85 Lower 95% CL 95.40
 “x mL” of standard stock solution 97.33 Upper 95% CL 99.75
Average (n=12) [%] 97.75
added RSD [%] 0.70
95% CI of Average ± 0.44%
 Samples withdrawn after 45
minutes, filtered and assayed
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Accuracy - Example

 Derived from Linearity Experiments: Slope Ratio

Standard Standard spiked into

formulation matrix
Linear slope: 11.1905 11.1540
y intercept: -7.0650 -7.5006
Correlation coefficient 0.9999 0.9995
Slope ratio 99.67%
(spiked/standard)

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Accuracy - Concepts

 Interference of the drug substance by matrix

 Different assumptions depending on type of dosage form
– Excipients dissolved together with the drug substance – i.e., constant proportional relationship
– Excipients suspended and dissolved prior to dissolution of drug substance – i.e., constant
mass of excipients with increasing amount of drug substance (standard addition technique)
– Excipients poorly or not dissolved - i.e., insoluble matrix with eluting drug substance
 All concepts applicable provided justification given with reference to technology
of dosage form

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Repeatability

 Analytical finish
– Replicate measurement of n=9 e.g., three concentrations and three replicates of each
concentration
• Standard solution
• Spiked solution
• Standard addition solution
– RSD < 2%
 Dissolution method
– Well-characterized product

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Precision/Repeatability

No. Drug dissolved [%] of label claim after 30 minutes

200 mg 400 mg 600 mg 800 mg*

1 100.8 100.9 100.8 97.7 99. 0

2 104.1 98.6 103.0 97.7 96.8
3 103.9 102.9 100.5 104.6 100.7
4 102.5 100.2 102.5 105.1 103.4
5 104.5 100.9 102.3 101.5 100.5
6 103.4 100.0 101.2 98.3 99.2

Dosage strength [mg] 200 400 600 800 800

Number of values 6 6 6 6 6
Mean value [%] 103.2 100.6 101. 7 100.8 99.9
Standard deviation 1.36 1.41 1.02 3.43 2.20
Rel. std. dev. (%) 1.32 1.41 1.00 3.40 2.20
Min. value (%) 100.8 98.6 100.5 97.7 96.8
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Intermediate Precision/Ruggedness

 Evaluate the effect of random events on

– Dissolution results
– Variance of the individual results
 Across the range of product strengths
 At least 2 different analysts on different days
 The difference in the mean value of the dissolution results from one analyst to
the other does not exceed
– 10% at time points with less than 85% dissolved;
– 5% for those remaining time points above 85%
 Acceptance criteria may be product specific
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Robustness

 Assesses the effect of small deliberate changes to the dissolution conditions

 Performed on well-characterized product

Factor Low Level Mid Level High Level

Rotational Speed of 95rpm 100rpm 105rpm
baskets
Surfactant concentration 0.20% 0.25% 0.30%
Temperature 36°C 37°C 38°C
Concentration of Acid 0.9mM 1.0mM 1.1mM
Analyst A n/a B
Instrument 1 n/a 2
Dose strength 1 n/a 2

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Robustness - Analytical Method

HPLC
 Variation of the mobile phase composition
 Flow rate
 pH value
 Column type
 Separation temperature
 Wavelength
Spectrophotometric analysis
 Wavelength

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What Should be Used for Validation?

Attribute Comment <1092>

Linearity At least 5 different concentration levels STD / SP*

Range -20% of the lowest conc. up to +20% of the STD / SP*

(linear, accuracy, precision) highest conc.

Accuracy/ recovery At least 3 different concentration levels SP*

Repeatability n=6 individual preparations WCDF

Intermediate precision Two analysts/one lab WCDF
Reproducibility Two analysts/two labs WCDF
Robustness Deliberate changes to parameters WCDF

STD = Standard solution;

SP = Spiked placebo;
WCDF = Well-characterized dosage form;
* = standard addition method is acceptable.
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Stability of Solutions

 Reference Standard solution

 Analyzed over a specified period of time
 Compared to a freshly prepared standard solution at each time interval
 Results between 98% and 102% of the initial value

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Stability of Solutions

 Sample solution
 Stored at room temperature
 Analyzed over a specified period of time
 Results compared to the initial results
 Results between 98% and 102% of the initial value
 Light protection and/ or cooling of the solutions may be required
 The method should state the expiry date of the solutions
 Use of glass containers instead of plastic may be necessary

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Consideration for Automation

 Deviations from USP <711> due to automation should be validated

– Resident sampling probe
– Sampling through the stirring element shaft
– Fiber optic probes
 Aspects specific to automation
– Carryover of residual drug
– Effect of an in-residence probe
– Adsorption of the drug in tubing, fittings, etc.
– Cleaning/ rinsing cycles
 Manual and automated procedures should be compared to evaluate
interchangeability
– n=12 for each manual and automated procedure
– Requirements for the intermediate procedure may apply
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Lessons Learned – Module 6

 Validation concept of combined steps

– The dissolution step
– Sampling and filtration
– Sample transfer
– Analytical finish

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(301) 230-6304 | [Link] | Education@[Link]
Module 03: Method
Development
Instructor Name

USP Affiliation: Position,

Committee
Title: Professional Title
Company: Company Name
Education: Degrees
Instructor Photo Degrees continued
Instructor bio…Natet pe volorib usapidus etur ad estruption porro ventorem
doluptibusam dempore con etur, esecear uptatus eum rem volupta doluptatur? Ectet
eos sit aut am vid quaes re dolum unt voluptatem fugia dolup. ad estruption porro
ventorem doluptibusam dempore con etur, esecear uptatus eum rem volupta
doluptatur? Ectet eos sit aut am vid quaes re dolum unt voluptatem fugia dolup.
Natet pe volorib usapidus etur ad estruption porro ventorem doluptibusam dempore con
Instructor bio continued if more space needed.. Natet pe volorib usapidus etur ad estruption porro ventorem
doluptibusam dempore con etur, esecear uptatus eum rem volupta doluptatur? Ectet eos sit aut am vid quaes re dolum
unt voluptatem fugia dolup. ad estruption porro ventorem doluptibusam dempore con etur, esecear uptatus eum rem
volupta doluptatur? Ectet eos sit aut am vid quaes re dolum unt voluptatem fugia dolup.
Natet pe volorib usapidus etur ad estruption porro ventorem doluptibusam dempore con etur, esecear uptatus eum rem
volupta doluptatur? Ectet eos sit aut am vid quaes re dolum unt voluptatem fugia dolup. ad estruption porro ventorem
doluptibusam dempore con etur, esecear uptatus eum rem volupta doluptatur? Ectet eos sit aut am vid quaes re dolum
unt voluptatem fugia dolup.
2
© 2018 USP
Disclaimer

Because USP text and publications may have legal implications in the U.S. and elsewhere, their language must
stand on its own. The USP shall not provide an official ex post facto interpretation to one party, thereby placing
other parties without that interpretation at a possible disadvantage. The requirements shall be uniformly and
equally available to all parties.
In addition, USP shall not provide an official opinion as to whether a particular article does or does not comply
with compendial requirements, except as part of an established USP verification or other conformity
assessment program that is conducted separately from and independent of USP's standard-setting activities.
Certain commercial equipment, instruments or materials may be identified in this presentation to specify
adequately the experimental procedure. Such identification does not imply approval, endorsement, or
certification by USP of a particular brand or product, nor does it imply that the equipment, instrument or material
is necessarily the best available for the purpose or that any other brand or product was judged to be
unsatisfactory or inadequate.
This course material is USP Property. Duplication or distribution without USP’s written permission is prohibited.
USP has tried to ensure the proper use and attribution of outside material included in these slides. If,
inadvertently, an error or omission has occurred, please bring it to our attention. We will in good faith correct
any error or omission that is brought to our attention. You may email us at: legal@[Link].
3
© 2018 USP
Learning Objectives – Module 3

 Dissolution method parameters which may have an influence on the dissolution

behavior of the dosage form
 Proper selection of dissolution testing conditions
 Assessment of the discriminatory property of a dissolution method

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Outline – Module 3

 Method development
– De-aeration
– Sinkers
– Agitation
– Study design
• Time points
• Observations
• Sampling
• Cleaning
– Data handling
– Dissolution procedure assessment

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Dissolution Testing

 Dissolution assesses the performance of drug products

 To be effective, the test should be:
– Predictive
• Relationship to in vivo response
– Comparative
• Prediction only possible with comparative tests
– Discriminatory
• Comparison only possible with discriminatory tests
– Reproducible
• Discrimination only possible with reproducible tests
– Precise
• Significant differences are based on the variability of the results
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Discriminating Dissolution Test

 Procedure should be appropriately discriminating, i.e., capable of distinguishing

relevant changes in product characteristics that might affect in vivo performance
 In vitro procedure may show differences between batches when in vivo
bioequivalence is demonstrated
 Need careful evaluation of whether the procedure is or appropriately
discriminating or too sensitive
 By intentionally varying manufacturing parameters (i.e. lubrication, blend time,
compression force, etc.) or stability testing parameters (temperature, humidity,
time) the discriminatory power of the procedure can be evaluated
 This information should be acquired during product development

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Dissolution Test - Variability of Results

 High variability in results can make it difficult to identify trends or effects of

formulation changes
 High variability caused by the test make it impossible to detect differences in
product quality
 Dissolution results may be considered highly variable if the relative standard
deviation (RSD) is greater than 20% at time points of 10 minutes or less and
greater than 10% at later
time points
 Most dissolution results exhibit less variability
 The source of the variability should be investigated and attempts should be
made to reduce variability of the
testing procedure
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Example of High Variability of Dissolution Results

V – vessel/sample
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Example of High Variability of Dissolution Results

V – vessel/sample

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Dissolution Test – Causes Of Artifacts

 Artifacts associated with the test procedure

– Coning
– Tablets sticking to the vessel wall or basket screen
– Buoyancy
– Any time the dosage contents do not disperse freely throughout the vessel in a uniform
fashion, aberrant results can occur
• Reactions taking place at different rates during dissolution:
 Excipient interactions or interferences
• Film coatings (pellicle forming)
• Aged capsule shell (“cross-linking”)
• Secondary inclusions of drugs in excipients (“dead extraction”)
– Visual observations are often helpful for understanding the source of the variability and
whether the dissolution test itself is contributing to the variability
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Dissolution Test – Decreasing Variability

 Usual remedies during method development include:

– Changing the apparatus type, speed of agitation, or de-aeration
– Consideration and/or examination of sinker type
– Changing the composition of the medium
– Changing other parts of instrumentation
• Sinkers
• Probes
• Vessels (size, material)
– Modifications to the apparatus may also be useful, with proper justification and validation
• Caveat: modified instrument no longer pharmacopeial standard!

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De-aeration of Dissolution Medium

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Example Deaeration of Dissolution Medium

 USP Monograph Lamivudine and Zidovudine Tablets

 Test 1 Medium: 0.1 N hydrochloric acid; 900 mL, deaerated
 Apparatus 2: 75 rpm
 Time: 15 min
 Lamivudine response factor solutions: 0.167 mg/mL of USP Lamivudine RS
in Medium. [NOTE—Prepare in duplicate.]
 Zidovudine response factor solutions: 0.333 mg/mL of USP Zidovudine RS
in Medium. [NOTE—Prepare in duplicate.]
 Sample solution: Pass a portion of the solution under test through a suitable
filter (PTFE, PVDF, or equivalent) of 0.45-µm pore size.
 Detector: UV 240–300 nm
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Sinkers

 Adjust the buoyancy of the dosage form that would otherwise float
 Keep the dosage form from sticking to the vessel wall
 Keep the dosage form in the same position
 Sinkers may influence the dissolution behavior
– Need detailed description in the method procedure
– Should have appropriate size
 Compendial sinker devices for App. 2

Source: [Link] last accessed on February 1, 2018

Source: [Link] last accessed on April 2, 2018
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Sinkers/Holders

 Compendial tablet holder for App. 4

 Non-compendial sinkers/holders

Source: [Link] last accessed on February 1, 2018

Source: [Link] last accessed on February 1, 2018
Source: [Link] last accessed on April 2, 2018
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Stationary Basket for Dosage Form Positioning

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Example – Specific Dissolution Testing Conditions USP Monograph
Efavirez Tablets

 Medium: 2.0% (w/v) sodium lauryl sulfate in water; 1000 mL. Do not deaerate.
 Apparatus 2: 50 rpm, with helix sinker. In addition, paddle and shaft must be
composed of stainless steel and not coated with Teflon or other material. Also,
all sampling devices and dissolution vessels must be washed with methanol or
ethanol followed by a water wash.
 Time: 30 min
 Sample solution: Pass a portion of the solution under test through a suitable
polyethylene filter, and dilute with Medium to obtain a concentration similar to
the Standard solution, assuming complete dissolution of the Tablet label claim.
 Instrumental conditions Analytical wavelength: UV 247 nm

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USP Monograph Levothyroxine Na Tablets

Test 4:

If the product complies with this test, the labeling indicates that it meets
USP Dissolution Test 4.[NOTE—Do not use paddle stirrers with synthetic coating.]
Medium: 0.01 N hydrochloric acid; 500 mL for Tablets labeled to contain between
25 and 175 µg of levothyroxine sodium;
and 900 mL for Tablets labeled to contain 200 or 300 µg of levothyroxine sodium
Apparatus 2: 75 rpm
Time: 45 min
Sample solution: Sample per Dissolution 〈711〉. Centrifuge the solution under
analysis.
Chromatographic system Mode: LC
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Common Agitation Rates and Temperature Values

 Basket Apparatus: 50 – 100 rpm

 Paddle Apparatus: 50 rpm – 75 rpm, above 100 rpm with proper justification
– Suspensions: 25 – 50 rpm
– Extended release formulations: 100 rpm
 Reciprocating Cylinder Apparatus: 5 dips/min – 35 dips/min
 Flow-through Cell Apparatus:
– 4 mL/min, 8 mL/min, 16 mL/min
– Variation of cell diameter with or without glass beads
• 12 mm, 22.6 mm diameter • Alternative flow-through cell types

 Temperature
– Most dosage forms: 37°C
– Dosage forms applied on the skin: 32°C
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Study Design – Time Points

Depends on the release pattern of the dosage form

 Immediate release dosage forms

– Development: profiles to characterize the dissolution pattern
(e.g., 10, 15, 20, 30, 45 and 60 min)
– Quality Control: meaningful reduction from profiles to single time point (e.g.,15 or 30 min)

 Orally disintegrating tablet formulation

– Early time points (e.g., < 5 min)

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Study Design – Time Points

 Modified release formulations

– Delayed release – enteric coated
• Acidic stage: 1 h or 2 h
• Buffer stage: similar to IR formulations
– Delayed release – not enteric coated
• Case by case
– Extended release formulations, at least
• Early time point: 1 or 2 hours
• Intermediate time point: 4 or 5 hours
• Final time point e.g., 8, 12 or 24 hours

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Additional Sampling for Method Development

Infinity Time Points may provide useful information about formulation

characteristics during initial development
 The paddle or basket speed is increased at the end of the run for a sustained
period (typically 15 to 60 minutes), after which time an additional sample is
taken
– No requirement for 100% dissolution in the profile

 Provides data that may supplement content uniformity data

 Provide information on stability of the dissolved drug substance

 Not required for routine testing

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Example: Infinity Time Point Evaluation

50 rpm 200 rpm

100

90

80
dissolution % of label claim

70

Mean
60
Vessel 1
50 Vessel 2
Vessel 3
40
Vessel 4
30 Vessel 5
Vessel 6
20

10

0
0 5 10 15 20 25 30 35 40 45 50 55 60 65
Time (min)

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Sampling – Consumptive or Non-consumptive

 Manual sampling
– Plastic or glass syringes
– Curved stainless steel cannula
– Filter
– Filter holder

 Automated sampling
 Sampling through basket or
paddle shaft
 In-situ using fiber optic

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Automated Sampling

 Cross contamination
 Drug adsorption
 Disturbance of hydrodynamics
 Carryover
 Rinsing and cleaning cycles

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Sampling Volume

The effect of withdrawn sample volume should be considered

 Profiling
– Removal of undissolved particles
– Reduction of bulk medium volume
• Impact on concentration gradient -> dissolution rate
– Loss of mass and loss of volume to be validated for:
• No replacement + no correction
• No replacement, but arithmetical correction
• Replacement + no correction
• Replacement + arithmetical correction

Not critical in case of

– App 4 when operated in open-system configuration
– App 3 when the time points correspond with changing rows
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Cumulative vs. Fractioned Data

Cumulative data display Fractioned data display

Sampling Time (min) Sampling Time (min)

Vessel Vessel
30 60 120 180 240 30 60 120 180 240

1 14.19 23.36 38.69 53.66 69.91 1 14.19 9.16 15.33 14.97 16.25

2 15.79 26.26 44.26 60.41 74.82 2 15.79 10.47 18.00 16.15 14.41

3 14.76 23.82 38.30 52.28 67.34 3 14.76 9.06 14.48 13.98 15.06

4 15.43 25.60 40.83 56.96 73.79 4 15.43 10.16 15.23 16.13 16.83

5 14.85 24.64 41.36 58.67 83.53 5 14.85 9.79 16.73 17.31 24.85

6 15.28 25.74 41.75 57.05 73.52 6 15.28 10.46 16.01 15.30 16.47

7 14.92 24.52 39.93 53.83 71.44 7 14.92 9.60 15.42 13.90 17.60

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Dissolution Procedure Assessment

 Sensitive to changes in critical quality attribute

– Intentionally change critical manufacturing parameters
– Stressed formulation

 Sufficiently rugged
 Reproducible

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 Example of Discriminating Dissolution Methods Used for Multisource
Product Comparison (1)

100 100
chloroquine phosphate dissolved /

chloroquine phosphate dissolved /

80 80

60 60
% of decl.

% of decl.
40 40

20 Resochin Tabletten, Bayer Vital, batch IT103J5 20 Resochin Tabletten, Bayer Vital, batch IT103J5
Chlorochin 250 mg Berlin-Chemie, Berlin-Chemie, batch 03003
Chlorochin 250 mg Berlin-Chemie, Berlin-Chemie, batch 03003
Weimerquin Tabletten, Biokanol Pharma, batch 9938600 U
Weimerquin Tabletten, Biokanol Pharma, batch 9938600 U
0 0
0 10 20 30 40 50 60 0 10 20 30 40 50 60
time / min time / min
chlqphos_water chlqphos_ph12

USP App. 2, 100 rpm USP App. 2, 75 rpm

900 mL water, 37ºC 500 mL SGF pH 1.2, 37ºC

Source: © 2006, Ph.D Thesis

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© 2018 USP
Example of Discriminating Dissolution Methods Used for Multisource
Product Comparison (1)

100 100
chloroquine phosphate dissolved /

chloroquine phosphate dissolved /

80 80

60 60
% of decl.

% of decl.
40 40

20 Resochin Tabletten, Bayer Vital, batch IT103J5

20 Resochin Tabletten, Bayer Vital, batch IT103J5
Chlorochin 250 mg Berlin-Chemie, Berlin-Chemie, batch 03003 Chlorochin 250 mg Berlin-Chemie, Berlin-Chemie, batch 03003
Weimerquin Tabletten, Biokanol Pharma, batch 9938600 U Weimerquin Tabletten, Biokanol Pharma, batch 9938600 U

0 0
0 10 20 30 40 50 60 0 10 20 30 40 50 60
time / min time / min
chlqphos_ph12 chlqphos_ph68

USP App. 2, 75 rpm USP App. 2, 75 rpm

500 mL SGF pH 1.2, 37ºC 500 mL SIF pH 6.8, 37ºC

Source: © 2006, Ph.D Thesis

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Example of Discriminating Dissolution Methods Used for Multisource
Product Comparison (1)

120 120

doxycycline dissolved / % of label claim

doxycycline dissolved / % of label claim

100 100

80 80

60 60

40 40
doxy 200 von ct, ct-Arzneimittel GmbH, batch B20499 doxy 200 von ct, ct-Arzneimittel GmbH, batch B20499
Doxycyclin STADA 200 mg Filmtabletten, STADApharm GmbH, batch 5611
Doxycyclin STADA 200 mg Filmtabletten, STADApharm GmbH, batch 5611
20
Azudoxat 200 mg, Azupharma GmbH & Co., batch 11608
Doxy-Diolan 200, BRAHMS Arzneimittel GmbH, batch 0011
20 Azudoxat 200 mg, Azupharma GmbH & Co., batch 11608
Doxy-Wolff 200, DR. AUGUST WOLFF Arzneimittel GmbH, batch 106010 Doxy-Diolan 200, BRAHMS Arzneimittel, batch 0011
Doxy-Wolff 200, DR. AUGUST WOLFF Arzneimittel, batch 106010

0 0
0 10 20 30 40 50 60 70 80 90 100 110 120 130 0 10 20 30 40 50 60 70 80 90 100
time / min time / min
doxycyc_water doxycyc_sif

USP App. 2, 75 rpm USP App. 2, 75 rpm

900 mL water, 37ºC 500 mL SIF pH 6.8, 37ºC

Source: © 2006, Ph.D Thesis

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Discriminating Dissolution Method for IR BCS Class II Soft Gelatin
Capsule Formulation (2)

Medium Solubility
pH 2 - 0.1 N HCl 484.27 μg/mL
pH 6.8 - phosphate buffer 4.32 μg/mL
pH 12.5 - buffer 3.16 μg/mL
Source: © 2016, Dissolution Technologies
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Dissolution Method of Multivitamin Soft-Gelatin Capsule Formulation
(*)
App. 2 with stationary basket method App. 3 method
120.0
120.0

Formulation A Formulation A
100.0
100.0

Riboflavin, % of assay value released

Riboflavin, % of assay value released

Formulation B
80.0
80.0
Formulation B

60.0
60.0

40.0
40.0

20.0
20.0

0.0
0.0 0 20 40 60 80 100
0 20 40 60 80 100
Time (min)
Time (min)

Two Multivitamin products, two batches each, color code retained

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Lessons Learned – Module 3

Experimental testing conditions depend on

 Physico-chemical properties of the  Physiology at the release site
drug substance – Gastrointestinal tract
– Aqueous solubility – Skin
– Stability
 Discriminating property of the
 Properties of the dosage form dissolution method is essential
– Rate of release
– Physical form
 Administration of dosages form
– Route of administration
– Time of administration

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References

1. Stippler ES, Biorelevant dissolution test methods, Ph.D Thesis, Shaker Verlag,
2006
2. Festo D, Marati M, Pathak V, Schwartzenhauer J, Development of a
discriminating dissolution method for immediate-release soft gelatin capsules
containing a BCS Class II compound, Diss. Techn. 23 Issue 4, 2016, 6-13

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(301) 230-6304 | [Link] | Education@[Link]