World Health Organization Training Workshop on Pharmaceutical Quality, GMP and Bioequivalence

Kiev - Ukraine 3 to 7 October 2005

Comparative dissolution testing and applications

Theo Dekker, D.Sc., consultant to WHO Research Institute for Industrial Pharmacy North-West University, Potchefstroom, South Africa iiftgd@puk.ac.za
Feb 2005

TG Dekker WHO, Malaysia

What is dissolution testing? tablets and capsules (conventional)

It measures the portion (%) of the API that (1) has been released from tablets/capsules and (2) has dissolved in the dissolution medium during controlled testing conditions within a defined period
The tablet thus first disintegrates Then the API will be able to dissolve Slow disintegration slow dissolution The % API dissolved is determined with an appropriate validated method: UV/VIS, HPLC, AA, GC, etc

Dissolution testing is also applicable to suspensions and suppositories

Feb 2005

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Glossary Solid oral dosage forms Immediate release typically means that 75% of the API is dissolved within 45 minutes
Rapidly dissolving: 85% in 30 minutes Very rapidly dissolving: 85% in 15 minutes Not part of presentation Modified-release dosage forms (consult Int.Ph., BP, USP)
Formulation deliberately changes release (slows down)
Extended-release (prolonged-release) Slower release throughout the GI tract Delayed-release (enteric coated tablets) Resists gastric fluid & disintegrates in intestinal fluid
Feb 2005

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What is multi-point dissolution?

In multipoint dissolution
multiple ( 3) samples are withdrawn from the dissolution medium during dissolution testing at pre-determined time points and each sample is analysed for the % API dissolved A graph of % API dissolved against time: The dissolution profile

Feb 2005

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Multi-point dissolution Example of dissolution profile

ACTIVE INGREDIENT: CLARITHROMYCIN MEDIUM: PHOSPHATE BUFFER pH 6.8
120

100

Dissolution (%)

80

60

40

20

Clarithromycin 250 mg tablets

0 0 10 20 30 40 50

WITHDRAWAL TIME IN MINUTES

Feb 2005

TG Dekker WHO, Malaysia

Comparative dissolution testing The principle

Two or more products or batches containing the same API are compared The strength of products / batches may or may not be the same (depending on purpose of test) The dissolution conditions are similar, e.g.
Apparatus, medium, volume, rotation speed & temp. Minimize possible experimental differences in conditions

Samples are taken at the same time points and the data (dissolution profiles) compared Calculations: correct for volume change of dissolution medium
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Feb 2005

Comparative dissolution testing Profile similarity determination Two conditions to determine if the dissolution profiles of two products/batches in a particular dissolution medium are similar:
1. If both the test and reference product show more than 85% dissolution within 15 minutes, the profiles are considered to be similar No calculations are required If this is not the case, apply point 2 2. Calculate the f2 value (similarity factor): If f2 50, the profiles are normally regarded similar
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Feb 2005

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Comparative dissolution testing Similarity factor f2

n = number of time points R(t) = mean % API dissolved of reference product at time point x T(t) = mean % API dissolved of test product at time point x Minimum of 3 time points (zero excluded) 12 units (each in own dissolution vessel) for each product (for official purposes) Only one measurement should be considered after both products have reached 85 % dissolution RSD at higher time points 10%
Feb 2005

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Comparative dissolution testing Dissolution conditions (study design)

Apparatus (choice) Paddle, 50 (75) rpm Basket, 100 rpm or

Dissolution media
All three media for full comparison Volume of media

1. Buffer pH 6.8 or simulated intestinal fluid without enzymes 2. Buffer pH 4.5 3. 0.1 M HCl or buffer pH 1.2 or simulated gastric fluid without enzymes 900 ml or less 37C 0.5C
10, 15, 20, 30, 45, (60, 120) min. (typical) 12 for official studies

Temperature
Sampling points Units (individual)

Feb 2005

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Typical time points Immediate release tablets (capsules)

Point Time
1 2 3 4 10 15 2. Condition 2, for calculation of f2 20 30
a minimum of 3 points are required Only one measurement should be considered after 85 % dissolution (both tablets) 20 minute time point thus first possible one (if 15 minute fails 1st condition) 10
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Rationale: 1. Condition 1

85% dissolution of both products within 15 minutes 15 minute time point thus essential

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Comparative dissolution testing Comparison of products When are dissolution properties of two products (batches) regarded similar?
The profiles should be similar in all three media
Statements of instability or insolubility are not acceptable, but should be demonstrated / justified

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Example 1 Determination of similarity of profiles

Example 1-A % API dissolved Example 1-B

Time Tablet A Tablet B (min) (Ref) (Test) 10 87 94 15 96 99 20 99 99 30 100 99 45 101 99 60 101 99 f2 required? No, 85% in 15 min f2 (n = N/A ?) profiles similar
Feb 2005

% API dissolved Time Tablet D Tablet E (min) (Ref) (Test) 10 55 57 15 72 78 20 85 91 30 97 100 45 102 100 60 103 101 f2 required? Yes f2 (n = 3 ?) 64 (similar)
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Example 1 Determination of similarity of profiles (cont.)

Example 1-C Example 1-D

% API dissolved Time Tablet X Tablet Y (min) (Ref) (Test) 10 29 34 15 38 41 20 47 50 30 63 64 45 80 79 60 95 91 f2 required? Yes f2 (n = 6 ?) 74 (similar)
Feb 2005

% API dissolved Time Tablet A Tablet Y (min) (Ref) (Test) 10 87 55 15 96 72 20 99 85 30 100 97 45 101 102 60 101 103 f2 required? Yes f2 (n = 3 ?) 31 (not similar)
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Example 2 Ciprofloxacin: two batches of same product

Product Cipro 500 Manufacturer Batch Nr Expiry date ABC Ltd xxx 06/2007 Status Test

Cipro 500
Apparatus Medium 1: Medium 2: Medium 3: Temp.: Units: Sampling: Analysis:
Feb 2005

ABC Ltd

zzz

07/2007

Reference

paddle at 50 rpm simulated gastric fluid without pepsin (SGF) (900 ml) acetate buffer pH 4.5 (900 ml) phosphate buffer pH 6.8 (900 ml) 37C 0.5C (start, middle, end) Twelve tablets per medium, each batch Manual, through in-line filter (0.45 m PVDF unit) at 10, 15, 20, 30 and 45 minutes HPLC analysis for ciprofloxacin 14
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Example 2 Ciprofloxacin: two batches (cont.)

Medium SGF, pH 1.16 % dissolved Buffer pH 4.5 % dissolved Buffer pH 6.8 % dissolved

Time (min) b/n xxx b/n zzz b/n xxx b/n zzz b/n xxx b/n zzz 10 83 80 93 96 28 31
15 20 95 99 92 97 97 99 99 100 34 38 36 39

30 45 similarity ? n= 5 ?

102 101 102 102 85% in 15 min.

100 100 102 101 85% in 15 min.

39 39 ( 50)

40 41

f2 = 83

Conclusion: The profiles in all three media can be regarded similar / not similar, since
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Example 2 Ciprofloxacin: two batches (cont.)

SGF without pepsin, pH 1.16
ACTIVE INGREDIENT: CIPROFLOXACIN MEDIUM: SGF WITHOUT PEPSIN pH 1.16

Acetate buffer pH6.8

ACTIVE INGREDIENT: CIPROFLOXACIN MEDIUM: ACETATE BUFFER pH 4.5

120

120 100

100

Dissolution (%)

Dissolution (%)

80

80 60 40 20 0

60

40

BATCH NO: zzz BATCH NO: xxx

BATCH NO: zzz BATCH NO: xxx

20

0 0 5 10 15 20 25 30 35 40 45 50

10

15

20

25

30

35

40

45

50

WITHDRAWAL TIME IN MINUTES

WITHDRAWAL TIME IN MINUTES

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Example 2 Ciprofloxacin: two batches (cont.)

Phosphate buffer pH 6.8
ACTIVE INGREDIENT: CIPROFLOXACIN MEDIUM: PHOSPHATE BUFFER pH 6.8

120

Dissolution (%)

100

BATCH NO: zzz

80 60 40 20 0 0 5 10 15 20 25 30 35 40 45

BATCH NO: xxx

WITHDRAWAL TIME IN MINUTES

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Ciprofloxacin (cont.)
Solubility is pH dependent:
Highly soluble at pH < 6 100% dissolution obtained in pH 4.5 and pH 1.16 At pH 6.8 and 40C the solubility is about 0.2 mg/ml Explains 40% dissolution for 500 mg dose !!
40C

Questions:
1. What dissolution level should ciprofloxacin 250 mg tablets be able to reach in pH 6.8 medium? 2. Should a change in particle size affect the dissolution rate?
X. Yu et al. Pharm. Research, 11, 522-527 (1994)
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Example 3
Lamivudine 150 mg & zidovudine 300 mg tablets
Source, WHO publication: Ongoing Monitoring of Antiretroviral Products as Part of WHOs Prequalification Project. Journal of Generic Medicines (accepted for publication, January 2006 edition) Samples from PQ project or bought/requested

Apparatus: Medium: Sample times: Analysis:

paddle at 75 rpm 900 ml 0.1 M hydrochloric acid, 37C 5, 10, 15, 20, 30 and 45 minutes HPLC

Data presented for individual APIs in next tables

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Example 3
Lamivudine 150 mg & zidovudine 300 mg tablets (2)
% Lamivudine of label claim dissolved Time (min) Combivir Gen-1 Gen-2 Gen-3 Gen-4

5 10 15 20 30 45 85 in 15 min ?
f2
Feb 2005

85 96 97 97 97 97 Reference

25 46 65 80 97 97 no
21 20

92 96 98 98 98 98

65 85 95 98 98 98

45 81 92 95 96 97

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Example 3
Lamivudine 150 mg & zidovudine 300 mg tablets (3)
% Zidovudine of label claim dissolved Time (min) Combivir Gen-1 Gen-2 Gen-3 Gen-4

5 10 15 20 30 45 85 in 15 min ?
f2
Feb 2005

85 97 98 98 98 99 Reference

22 44 64 81 100 100 no
20 21

74 90 97 99 101 100

68 88 96 100 99 99

45 83 95 98 99 100

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Example 3
Lamivudine 150 mg & zidovudine 300 mg tablets (4)

Conclusion (considering only 0.1 M HCl as medium)

1. 3 products show profile similarity with Combivir ( 85% in 15 minutes) 2. The profiles of Combivir and Gen-1 are not similar
The products may still show bio-equivalency

The dissolution profiles of the APIs in a particular product are similar (true for all 5 products) Examples: see profiles of Combivir and Gen-1
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Example 3
Lamivudine 150 mg & zidovudine 300 mg tablets (5)
120

120
100

Dissolution (%)

80 60 40 20 0 0 10 20 30 40

Dissolution (%)

100 80 60 40 20 0

Lamivudine Zidovudine

Lamivudine Zidovudine

50

10

20

30

40

50

Time (minutes)

Time (minutes)

Combivir dissolution profile 0.1 M hydrochloric acid

Gen-1 dissolution profile 0.1 M hydrochloric acid

Note the similarity of the API profiles of each product APIs highly soluble = dissolution controlled by disintegration time Is particle size of APIs expected to be critical ?
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Example 3
Clarithromycin tablets Proportional formulations
2 strengths prepared from same granulate f2 = 31 Profiles not similar ! Solubility of the API in buffer pH 6.8 low according to BCS Do you expect that particle size or polymorphism may have effect on the profiles?
ACTIVE INGREDIENT: CLARITHROMYCIN MEDIUM: PHOSPHATE BUFFER pH 6.8
120 100

Dissolution (%)

80 60 40 20 0 0 10 20 30 40 50

PRODUCT B 500 mg PRODUCT B 250 mg

WITHDRAWAL TIME IN MINUTES

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Effect of Particle Size on Dissolution of Nevirapine Tablets (Source: Ranbaxy)

USP Type II / 0.01N HCl 50 RPM / 900 ml 120

% Drug Dissolved

1 Viramune 2 Nevipan 90% < 31 3 Nevipan 90% < 81 f2 : 1 vs 2 = 72 f2 : 1 vs 3 = 31 X f2 : 2 vs 3 = 34 X

100 80 60 40 20 0

1 2 3

Time (Min) Nevipan MGS(1024)05 (90%LT81.12) Nevipan MGS(1024)60B (90%LT30.89) Viramune 992633B

0 0.00 0.00 0.00

10 50.80 80.00 83.30

20 73.80 92.00 96.60

30 83.98 96.00 97.70

http://www.who.int/medicines/organization/par/FDC/VKAroraWHOGenevaDec.ppt
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Applications
1. For selection of the formulation in the development phase

By comparison of the dissolution profiles of innovator product with those of formulations Hint: start with comparator product to see:
Immediate release? Rapidly dissolving? Very rapidly dissolving? Disintegration testing can aid in the early phases

This should be a basic strategy in R&D to maximize the chances of bioequivalence

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Feb 2005

Applications (cont.)
2. It is a requirement of the prequalification programme to submit comparative dissolution data for the bio-batch and innovator batch
Same batches as used in bioequivalence study ! Submit report with data, profile comparison & discussion (see report requirements) This report form part of pharmaceutical development report
Inclusion of the same report in the bioequivalence study report is recommended 27
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Feb 2005

Applications (cont.)
3. Demonstration of in vivo bioequivalence of one or more of the lower strength(s) of an FPP may be waived based on
1. an acceptable in vivo BE study of the highest strength against the comparator product 2. demonstration of similarity of dissolution profiles, 3. if the lower strength is proportionally similar in formula to the higher strength (bio-batch) and 4. if all pharmacokinetic requirements are met
Consult the bio-guideline, also for reverse situation

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Applications (cont.)
4. Comparison of the release properties of pivotal batches
To demonstrate in vitro similarity of such batches
This is considered essential for retention of efficacy and safety Note that bioequivalence studies are done normally only once on a bio-batch during development It must be demonstrated that the product retains the dissolution characteristics up to production scale

The studies should be submitted in dossier as part of the FPP development report
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Applications (cont.)
5. Selection of the dissolution specifications for product release & stability purposes
1. Conditions and acceptance criteria to be set 2. The dissolution profiles of the bio-batch should be used for this purpose 3. A dissolution specification should be able to detect inadequate release properties of the commercial batches
A generous dissolution limit has no quality selectivity 4. Example: Combivir (from limited data in Example 3) 80% (Q) within 20 (15?) minutes for both APIs under conditions described in Example 3
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Applications (cont.)
6. Post-approval amendment application
A requirement of a particular change may be to demonstrate that the profiles of the amendment batch and the current batch are similar Consult guideline on variations

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Reporting Comparative dissolution data Full report, including

1. Purpose of study 2. Products / batches information
Batch number, manufacturing/expiry date, packaging, etc. CoA & size for own batches (and BMR for bio-studies report)

3. Dissolution conditions and method 4. Analytical method or reference to part of dossier 5. Results (% API dissolved)
Tabulated Graphically Similarity determination / calculation

6. Conclusion
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Guidelines
WHO Prequalification
1. Supplement 1 [for use from July 2005 (CPH25)] to: Guideline on Submission of Documentation for Prequalification of Multi-source (Generic) Finished Pharmaceutical Products (FPPs) Used in the Treatment of HIV/AIDS, Malaria and Tuberculosis Dissolution testing Others
Guidance for Industry. Waiver of In-Vivo Bioavailability and Bioequivalence Studies for Immediate-Release Solid Oral Dosage Forms Based on a Biopharmaceutics Classification System. U.S. Department of Health and Human Services, Food and Drug Administration, Center for Drug Evaluation and Research (CDER), August 2000. CPMP Note for Guidance on the Investigation of Bioavailability and Bioequivalence. The European Agency for the Evaluation of Medicinal Products CPMP/EWP/QWP/1401/98, July 2001

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Some conclusions
Comparative dissolution
should form an essential part of R&D of solid oral dosage forms (including suspensions), supports bio-studies, is required for comparison of pharmaceutical release properties of pivotal batches, is used to set dissolution specifications, and assists in post-approval changes

It is thus important

Feb 2005

to conduct the studies under controlled conditions in the 3 media, all as required by the guidelines, to take samples for analysis at meaningful intervals and to be able to determine similarity of profiles
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