IBCs 7th International Conference on

Stability Testing
How to Select, Validate and Report
Analytical Test Procedures Used
to Monitor Stability
Jean Gilliard
Forte Posthouse Regent Park, London 6&7 February 2001

SGS Lab Simon S.A.

Test procedures
Selection of the tests:
The testing should cover the features susceptible
to change during storage and likely to influence
quality, safety and efficacy.

Validation of the tests:

Validated stability-indicating analytical procedures
should be applied

SGS Lab Simon S.A.

Selection of the test procedures

Stability testing should cover as necessary the
organoleptic, physical, chemical and microbiological
test characteristics.
In general, appearance, assay and degradation
products determination should be evaluated
whatever the dosage form.
Considerations for specific dosage forms

SGS Lab Simon S.A.

Tablets & Capsules

Physico-chemical stability
appearance, average weight, moisture, hardness,
friability, disintegration time, dissolution rate,
assay and degradation products determination

Microbiological stability
microbial limits

SGS Lab Simon S.A.

Creams & Ointments

Physico-chemical stability
appearance, particle size, viscosity, consistency,
pH, preservative content (if present), assay and
degradation products determination

Microbiological stability
microbial limits, preservative effectiveness

SGS Lab Simon S.A.

Injections
Physico-chemical stability
appearance, particulate matter, pH,
preservative content (if present), assay and
degradation products determination

Microbiological stability
sterility,pyrogenicity, preservative effectiveness

SGS Lab Simon S.A.

Stability-Indicating Methodology
Validated quantitative analytical methods that
can detect the changes with time in the chemical,
physical or microbiological properties of the drug
substance or product and that are specific so that
the contents of active ingredient, degradation
products, and other components of interest can be
accurately measured without interference.

SGS Lab Simon S.A.

Stability-Indicating Methodology
Requirements
3high specificity
3high sensitivity
3high precision

SGS Lab Simon S.A.

Stability-Indicating Methodology
Quantitative
Technique
Titrimetry

poor

poor

high

Universal
detection
capability
poor

UV-vis
spectrometry

poor

poor

high

good -

HPLC

good

good

acceptable

good

CE

good +

acceptable

acceptable

good +

SGS Lab Simon S.A.

Specificity Sensitivity

Precision

Alternative procedures (Q6A)

For formulation shown to be stable during
manufacture, non separative techniques may
be used at release with the chromatographic
method as the official procedure
When conclusively demonstrated with a
significant body of data that the formulation is
stable, degradation product testing may be
reduced or eliminated
SGS Lab Simon S.A.

ICH validation requirements (Q2A)

Attribute

Assay

Related substances

Specificity

LOQ

Linearity

Accuracy

Precision
Repeatability
Intermediate Precision

+
+

+
+

SGS Lab Simon S.A.

Specificity
For the main component(s)
For the related substances (impurities and
degradation products)
Supported by stress testing studies

SGS Lab Simon S.A.

Stress testing
Conducted to demonstrate the stability
indicating power of the analytical procedures
Should be performed on the drug substance
and on the drug product
Should included the effects of temperature,
moisture, pH, oxidation and light
Usually aim to achieve 10 to 20 % degradation

SGS Lab Simon S.A.

Stress testing
Results from stress testing studies should
form an integral part of the information
provided to regulatory authorities
Attention should be paid to reviewing the
adequacy of the mass balance

SGS Lab Simon S.A.

Mass balance
The process of adding together the assay values
and levels of degradation products to see how
closely these add up to 100 % of the initial value
with due consideration of the margin of analytical
error
Good mass balance (assay + degradants = 100 %)
can support validation
Poor mass balance can suggest an analytical
problem
SGS Lab Simon S.A.

Mass balance
HPLC may fail to detect some components

retained on the column

eluted late
eluted under main peak
eluted at solvent front
eluted undetected

SGS Lab Simon S.A.

Mass balance
Extraction complete ?
Response factors of the secondary
peaks ?

SGS Lab Simon S.A.

Stress testing
Should included consideration for excipients
effect
Tablet
Tabletstress:
stress:44weeks
weeksatat60C
60C

API +

SGS Lab Simon S.A.

Stress testing
Should included consideration for container
effect
Spectrum Index Plot

Estradiol - 16.211
250.00
nm
300.00

250.00

16.21

Peak16 - 17.756
nm
300.00

260.3

a-Estradiol - 19.041
250.00
nm
300.00

17.75 221.5

19.04

Syringes
Syringesfilled
filledwith
withgel
gel
stress:
stress:24
24hours
hoursatat
40C/75%RH
40C/75%RH
280.4

280.4

319.5 343.3

315.9

345.7

Estradiol - 16.211

0.30

0.25

0.15

Peak16 - 17.756

0.10

0.05

a-Estradiol - 19.041

AU

0.20

0.00
5.00

SGS Lab Simon S.A.

10.00

15.00

20.00
Minutes

25.00

30.00

SampleName In Use/1 Vial 24 Injection 1 Date Acquired 12/21/00 2:32:12 AM

35.00

40.00

Sensitivity: quantitation limit

Confidence that the method is capable of
providing the level of sensitivity required
LOQ reporting threshold (Q3B)
Maximum Daily Dose

Reporting Threshold

= 1g
> 1g

0.1 %
0.05 %

SGS Lab Simon S.A.

Quantitation Limit (Q2A&B)

LOQ: the lowest amount of analyte which can be
quantitatively determined with suitable precision and
accuracy
Approaches:
SNR: 10:1
SD of response and slope: 10 x / S
S = slope of calibration curve,
= SD of blank, RSD of regression line, SD of intercept

The estimate should be validated by the analysis of a

suitable number of samples known to be near or
prepared at the LOQ
SGS Lab Simon S.A.

Precision
Analysis repeatability
Intermediate precision: establish the effect
of variations e.g. different days, analysts,
equipment,
Reproducibility: interlaboratory trial

SGS Lab Simon S.A.

Precision (Q2B)
Repeatability:
a minimum of 9 determinations over the range of
the procedure (e.g. 3 replicates at 3 concentrations)
or a minimum of 6 determinations at 100 % of the
test concentration

Recommended data: SD, RSD and confidence

intervals
SGS Lab Simon S.A.

Precision
HPLC level of precision
Repeatability: 0.5 to 1 %
Intermediate precision: 1 to 2 %
Reproducibility: 2 to 5 %

Adequate to detect 2 to 3 % potency loss

over a three year storage period ?

SGS Lab Simon S.A.

Reporting Stability Data

Test Methodology Information:
Physical, chemical and microbiological
attributes and regulatory specifications
Test methodology used
Information on accuracy, precision and
suitability of the methodology

SGS Lab Simon S.A.

System suitability
Ensure method performs in-use as at the time of
validation
Link between validation and routine testing
Selection of relevant parameters from validation
Setting of acceptance limits
Stability-indicating methodology: check adequate
resolution, quantitation limit and precision at the
time of use
SGS Lab Simon S.A.

Method transfer
To assure the integrity of the analytical data in
recipient laboratory
To assess competency in recipient laboratory
To demonstrate the adequacy of equipment in
recipient laboratory
To identify limitations of the method
Ensure good communication between development
and recipient laboratories

SGS Lab Simon S.A.

Summary
Assays and related substance tests must be fully validated and
demonstrate stability indicating
Specificity, sensitivity and precision are the most crucial
attributes for a stability indicating methodology
System suitability testing must be carried out to demonstrate
suitable method performance at the time of use
Consider other tests which may be useful for monitoring
product stability

SGS Lab Simon S.A.