1. Integration of Forced Degradation Study in
LC Stability Indicating Method Development
Katy Dewitte, Ph.D. Pharm
Knowledge Manager, Anabiotec
1

2. Introduction Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
Today’s Agenda
 Forced degradation: definition and principles
 The stability indicating power of an analytical method
 Experimental design to conduct a study
 Practical example: case study
 Challenging the stability indicating method
 Conclusion
2

3. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
Definition of forced degradation or stress testing
The process of subjecting drug compounds:
DRUG SUBSTANCE
DRUG PRODUCT
PLACEBO
…
to extreme chemical and environmental conditions.
3

4. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
3 main purposes (according to ICH and FDA):
 To provide a stability assessment of the drug substance (DS) or
drug product (DP), to determine the intrinsic stability of a molecule
 To elucidate the possible degradation pathways of the DS or the
API in the DP (by identifying the likely degradation products)
 To investigate the stability indicating power of the analytical
method applied for DS and DP.
4

5. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
A stability indicating method:
 is a validated, quantitative analytical procedure
 can detect the changes with time in the pertinent properties of the
drug substance and drug product; is able to detect the increase in
degradation products.
 measures accurately the active ingredients without interference
from degradation products, process impurities, excipients, or
other potential impurities (specificity)
5

6. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
Development of a stability indicating method:
Purpose of forced degradation studies:
 Formation of potential degradation products
 Purposeful degradation
 Which degradation products can be expected during storage and
stability studies?
Need for appropriate forced degradation studies!
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7. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
4 main degradation mechanisms:
 Heat (thermolytic degradation)
 Hydrolytic degradation
 Oxidative degradation
 Photolytic degradation
Degrade the drug by ± 10%, or slightly exceeding the accelerated
stability conditions
7

8. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
Drug Substance
Solid Solution /
Suspension
 Photolysis  Photolysis
 Heat  Acid/base hydrolysis
 Heat + humidity  Oxidation
8

9. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
Drug Substance
Solid
 Light, 1 X ICH
 Photolysis  Light, 3 X ICH
 Heat  Light control
 Heat + humidity
9

10. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
Drug Substance
Solid
More severe than accelerated
(40°C / 75%RH)
 Photolysis  60°C or 80°C
 Heat  1, 3 and 5 days
 Heat + humidity
10

11. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
Drug Substance
Solid
More severe than accelerated
(40°C / 75%RH)
 Photolysis  60°C / 75%RH or
aa80°C / 75%RH
 Heat
 1, 3 and 5 days
 Heat + humidity
11

12. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
Drug Substance
Solution /
Suspension
 Light, 1 X ICH
 Light, 3 X ICH  Photolysis
 Light control  Acid/base hydrolysis
 Oxidation
12

13. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
Drug Substance
Solution /
 Solutions at various pH (2-12) Suspension
 0.1M – 1M HCl
 0.1M – 1M NaOH  Photolysis
 Room temperature or elevated  Acid/base hydrolysis
 1, 3 and 5 days  Oxidation
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14. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
Drug Substance
Solution /
Suspension
 0.1% – 3% H2O2
 Metal ions
 Photolysis
 Radical initiators
 Acid/base hydrolysis
 1, 3 and 5 days
 Oxidation
14

15. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
Case: A new drug substance (early phase)
Step 1: Development of a (stability indicating?) method
0.060
0.050
2 .0 0
0.040
1 .8 0 0.030
0.020
1 .6 0 0.010
AU
0.000
1 .4 0
-0.010
-0.020
1 .2 0
-0.030
AU
-0.040
1 .0 0
-0.050
0 .8 0 -0.060
3. 00 4. 00 5.00 6.00 7.00 8.00 9.00 10.00 11.00 12.00 13.00 14.00 15.00 16.00 17.00
Minutes
0 .6 0
0 .4 0
0 .2 0
0 .0 0
0 .0 0 2 .0 0 4 .0 0 6 .0 0 8 .0 0 1 0 .0 0 1 2 .0 0 1 4 .0 0 1 6 .0 0 1 8 .0 0 2 0 .0 0
M in u t e s
15

16. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
Quality-by-design approach
Software based development of an analytical method
 DS
 Starting materials
 Degradants
 Synthesis impurities
 A stressed DS sample (if any information is available)
16

17. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
Quality-by-design approach
17

18. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
Quality-by-design approach
18

19. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
Quality-by-design approach
0.30
7 .3
0.28
0.26
0.24
0.22
0.20
0.18
0.16
5 .5 5 6
0.14
AU
4 .53 9
0.12
0.10 6 .5 9 3
0.08
0.06
11. 511
14. 666
0.04
4. 76 4
10. 057
5.2 49
7.9 07
0.02
0.00
-0.02
-0.04
1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 11.00 12.00 13.00 14.00 15.00
Minutes
19

20. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
Photostress: 1 X ICH and 3 X ICH
0 .0 3 5
0 .0 3 0
0 .0 2 5
0 .0 2 0
0 .0 1 5
0 .0 1 0
0 .0 0 5
3 x ICH-light
AU
0 .0 0 0
- 0 .0 0 5
- 0 .0 1 0 1 x ICH-light
- 0 .0 1 5
- 0 .0 2 0
T0
- 0 .0 2 5
- 0 .0 3 0
- 0 .0 3 5
3 .0 0 4 .0 0 5 .0 0 6 .0 0 7 .0 0 8 .0 0 9 .0 0 1 0 .0 0 1 1 .0 0 1 2 .0 0 1 3 .0 0 1 4 .0 0 1 5 .0 0 1 6 .0 0 1 7 .0 0
Min u te s
20

21. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
Photostress: 1 X ICH and 3 X ICH
0 .0 3 5
0 .0 3 0
0 .0 2 5
0 .0 2 0
0 .0 1 5
0 .0 1 0
0 .0 0 5
HT
LIG E 3 x ICH-light
AU
NOT SITIV
0 .0 0 0
- 0 .0 0 5
1 x ICH-light
SE N
- 0 .0 1 0
- 0 .0 1 5
- 0 .0 2 0
T0
- 0 .0 2 5
- 0 .0 3 0
- 0 .0 3 5
3 .0 0 4 .0 0 5 .0 0 6 .0 0 7 .0 0 8 .0 0 9 .0 0 1 0 .0 0 1 1 .0 0 1 2 .0 0 1 3 .0 0 1 4 .0 0 1 5 .0 0 1 6 .0 0 1 7 .0 0
Min u te s
21

22. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
Solid stress: 60°C 7 days ~ 60°C / 75%RH 7 days
0 .0 3 5
0 .0 3 0
0 .0 2 5
0 .0 2 0
0 .0 1 5
0 .0 1 0
0 .0 0 5
60°C / 75%RH
AU
0 .0 0 0
- 0 .0 0 5
- 0 .0 1 0 60°C
- 0 .0 1 5
- 0 .0 2 0
T0
- 0 .0 2 5
- 0 .0 3 0
- 0 .0 3 5
3 .0 0 4 .0 0 5 .0 0 6 .0 0 7 .0 0 8 .0 0 9 .0 0 1 0 .0 0 1 1 .0 0 1 2 .0 0 1 3 .0 0 1 4 .0 0 1 5 .0 0 1 6 .0 0 1 7 .0 0
Min u te s
22

23. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
Solid stress: 60°C 7 days ~ 60°C / 75%RH 7 days
0 .0 3 5
0 .0 3 0
0 .0 2 5
0 .0 2 0
0 .0 1 5
BLE / 75%RH
0 .0 1 0
STA
0 .0 0 5
60°C
MO
AU
0 .0 0 0
H ER
- 0 .0 0 5
- 0 .0 1 0
- 0 .0 1 5
T 60°C
- 0 .0 2 0
T0
- 0 .0 2 5
- 0 .0 3 0
- 0 .0 3 5
3 .0 0 4 .0 0 5 .0 0 6 .0 0 7 .0 0 8 .0 0 9 .0 0 1 0 .0 0 1 1 .0 0 1 2 .0 0 1 3 .0 0 1 4 .0 0 1 5 .0 0 1 6 .0 0 1 7 .0 0
Min u te s
23

24. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
Hydrolysis: Acid: 0.1 M HCl 1 and 3 days
0.035
0.030
0.025
0.020
0.015
0.010
0.005
0.1M HCl; 3 days
AU
0.000
-0.005
-0.010 0.1M HCl; 1 day
-0.015
-0.020
T0
-0.025
-0.030
-0.035
3.00 4.00 5. 00 6.00 7.00 8.00 9.00 10. 00 11.00 12.00 13.00 14.00 15.00 16.00 17.00
Minutes
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25. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
Hydrolysis: Acid: 0.1 M HCl 1 and 3 days
0.035
API Degr. 1
0.030
0.025
T0 99.3% 0.7%
0.020
T1d 78% 22%
0.015
0.010
T3d 45% 55%
E IN
0.005
0.1M HCl; 3 days
AB L
AU
0.000
N ST
-0.005
-0.010
U 0.1M HCl; 1 day
ACID
-0.015
-0.020
T0
-0.025
-0.030
-0.035
3.00 4.00 5. 00 6.00 7.00 8.00 9.00 10. 00 11.00 12.00 13.00 14.00 15.00 16.00 17.00
Minutes
25

26. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
Hydrolysis: pH 2: 1, 3 and 7 days; room temperature
0.035
0.030
0.025
0.020
0.015
0.010
pH 2.0; 7 days
0.005
pH 2.0; 3 days
AU
0.000
-0.005
-0.010 pH 2.0; 1 day
-0.015
-0.020
T0
-0.025
-0.030
-0.035
3.00 4.00 5. 00 6.00 7.00 8.00 9.00 10. 00 11.00 12.00 13.00 14.00 15.00 16.00 17.00
Minutes
26

27. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
Hydrolysis: pH 2: 1, 3 and 7 days; room temperature
0.035
API Degr. 1 Degr. 2
0.030
T0 98.9% 0.1% 0.9%
0.025
0.020 T1d 97.6% 0.3% 2.0%
0.015
T3dpH 2.0; 70.3%
92.2% days 7.4%
0.010
E IN
0.005 T7d 79.4% 0.4% 20.3%
pH 2.0; 3 days
AB L
AU
0.000
N ST
-0.005
-0.010
U pH 2.0; 1 day
ACID
-0.015
-0.020
T0
-0.025
-0.030
-0.035
3.00 4.00 5. 00 6.00 7.00 8.00 9.00 10. 00 11.00 12.00 13.00 14.00 15.00 16.00 17.00
Minutes
27

28. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
Oxidation: 0.03% H2O2 1 and 3 days; 25°C
0.035
0.030
0.025
0.020
0.015
0.010
0.005
0.03% H2O2; 3 days
AU
0.000
-0.005
-0.010
0.03% H2O2; 1 day
-0.015
-0.020
T0
-0.025
-0.030
-0.035
3.00 4.00 5. 00 6.00 7.00 8.00 9.00 10. 00 11.00 12.00 13.00 14.00 15.00 16.00 17.00
Minutes
28

29. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
Oxidation: 0.03% H2O2 1 and 3 days; 25°C
0.035 API Degr. 1 Degr. 2 Degr. 3
T0 98.8% 0.0% 0.2% 0.0%
0.030
0.025
0.020 T1d 98.4% 0.1% 0.4% 0.3%
0.015
0.010
T3d 96.1% 0.2% 2.3% 0.6%
0.005
TO H2O2; 3 days
0.03%
IVE 0.03% H O ; 1 day
SIT
AU
0.000
E N EN
-0.005
-0.010
S 2 2
-0.015
-0.020
OX YG T0
-0.025
-0.030
-0.035
3.00 4.00 5. 00 6.00 7.00 8.00 9.00 10. 00 11.00 12.00 13.00 14.00 15.00 16.00 17.00
Minutes
29

30. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
Starting materials, intermediates and synthesis impurities
0.055
0.050
0.045
0.040
0.035
0.030
0.025
AU
0.020
0.015
0.010
Synthesis
0.005
impurities
0.000
-0.005
Drug Substance
-0.010
1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 11.00 12.00 13.00 14.00 15.00
Minutes
30

31. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
Stressed conditions
0.100
0.090
0.080
0.070
0.060
0.050
AU
0.040
Acid hydrolysis
0.030
0.020
Oxidation: metal ions
0.010 Oxidation: H2O2
0.000
Drug Substance
-0.010
1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 11.00 12.00 13.00 14.00
Minutes
31

32. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
Intermediates and synthesis impurities vs H2O2 stress
0.060
0.055
0.050
0.045
0.040
0.035
0.030
0.025
AU
0.020
0.015
Synthesis
0.010
impurities
0.005
0.000
-0.005
H2O2 stressed DS
-0.010
-0.015
1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 11.00 12.00 13.00 14.00 15.00
Minutes
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33. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
Conclusion of the case study
The method developed with the initial information is already very
suitable for conducting forced degradation experiments.
Full data from the 4 stress mechanisms indicate if additional method
optimization is needed.
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34. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Conclusion
Power Method
Conclusion of the case study
Consider the guidelines: only degradants that will be expected under
normal storage conditions must be taken into account.
Fit for purpose combined with Quality-by-design approach reduces the
workload and pass-through time.
34

35. Forced Degradation Stability Indicating Experimental Design Practical Example Stability Indicating Acknowledgement
Power Method
Acknowledgement
Mr. Herman Nuytten, CSO of AnaBioTec
Mrs. Ineke Verweire, COO of AnaBioTec
The project managers from AnaBioTec
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36. Questions?
36