ISSN 0975-6299 Vol.1/Issue-4/Oct-Dec.

2010

International Journal of Pharma and Bio Sciences

DEVELOPMENT OF ACCELERATED STABILITY PROTOCOL FOR

SILDENAFIL TABLETS – A EUROPEAN PERSPECTIVE
REVIEW

SUKHDEV SINGH*1 AND JASBIR SINGH2

1
Rayat Institute of Pharmacy, Punjab Technical University, S.B.S. Nagar, Punjab.
2
Rayat Institute of Pharmacy, Punjab Technical University, S.B.S. Nagar, Punjab.

*Corresponding Author [email protected]

ABSTRACT
Frequently, the goal of a pharmaceutical company is to develop a globally acceptable registration
stability protocol. A sound stability protocol not only eliminates unnecessary testing but also
reduces manufacturing needs, cost and time. In this article considerable issues related to
development of stability protocol such as type, size and number of batches, type, size and sources
of containers and closures, container closures orientation, sampling plan, storage conditions, test
time points, test parameters, test methods, acceptance criteria and the applicability of statistical
methods for the analysis of stability data is discussed. The aim of this paper is to develop and
outline accelerated stability protocol for Sildenafil tablets acceptable for registration in Europe and
highlight some of the considerations that must be made before the execution of actual stability
study.

KEYWORDS
Stability protocol; Sildenafil tablets; European Guidelines.

INTRODUCTION
protocol not only eliminates unnecessary
The stability of pharmaceutical ingredients and testing but also reduces manufacturing
the products containing them depends on two needs, cost and time. This is especially
major factors: important in current scenario due to increase

The chemical and physical properties of the in the number of possible storage conditions
materials concerned (including the and checkpoints because of stringent
excipients and container closure systems regional requirements2.
used for packaging of formulated products). The aim of this paper is to develop and

Environmental factors, such as temperature, outline important characteristics of a stability
humidity and light and their effect on the protocol acceptable for registration in Europe
drug products1. and highlight some of the considerations that
Frequently, the goal of a pharmaceutical must be made before the execution of actual
company is to develop a globally acceptable stability study. Here the general requirements
registration stability protocol. A sound stability for development of stability protocol and
stability testing of pharmaceuticals for

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registration in the European Community
For conventional dosage form, and when
(European Union) are discussed with respect to the active substance is known to be
the sources of information on requirements and stable, stability study should be carried
design of stability protocol for finished drug out on at least two pilot scale batches.
product3.
For critical dosage form, and when the
The protocol contains an outline of the active substance is known to be unstable,
proposed plan to be used in generating stability stability study should be carried on at
data. The protocol describes the type of product least three primary batches9.
being tested, sampling process, duration and For containers and closures ICH & European
frequency of testing, number of samples and guidelines refer stability study to be carried
replicates per time interval, storage conditions out on at least one container of each
(length of storage, type of storage, packaging material for testing where as US
temperatures and packaging), methods of refers sampling of at least two containers of
analysis with associated supportive data, if each packaging material for testing. FDA
available, and other tests4. draft guidelines set the frequency as 0, 2, 4
and 6 months for 6 months accelerated
The test protocol contains the following stability data to be submitted along with the
information application. WHO & EU guidelines, however,
1. Type, size and number of batches. suggest sampling at 0, 1, 2, 3 and 6 months
2. Type of containers and closures, Pack detail under accelerated conditions. For the
and pack size countries in zone I and II the storage
3. Sampling plan. conditions prescribed for long term testing are
4. Test storage conditions. 25 ± 2ºC and 60 ± 5% RH (ICH and FDA)
5. Test time points. whereas WHO guidelines suggest that
6. Test parameters conditions for long term studies should be as
7. Test methods close as to the derived conditions of 30 ± 2ºC
8. Acceptance criteria5. and 35 ± 5% RH for zone III countries and 30
± 2ºC and 70 ± 5% RH for zone IV
Although, all stability protocols contain a countries10. Consideration of all these
schedule for testing samples stored at one or stringent requirements pointed out only one
more controlled storage conditions, the protocol thing that it is utmost important to consider
can differ significantly from one product to regional requirements while developing
another and from country to country. If a drug stability protocol. Hence, when we developed
product is to be marketed solely in one country, the accelerated stability protocol for sildenafil
then it is necessary to design a straightforward tablets for European market, we considered
stability protocol on the basis of local following parameters:
regulations6. Although pharmaceutical
companies follow the ICH guidelines for the 1. Type, size and number of batches
stability testing but still there are some specific The guidelines emphasized the establishment
regional requirements of different countries of an expiration period that is based on
which must be considered for stability studies limited number of batches. The testing of
and hence for getting approval of drug product. three batches provides a reliable estimate
For example, for selection of batches ICH while testing fewer than three batches does
refers stability study to be carried out on at not permit a reliable statistical estimate of
least three primary batches of the drug product, batch to batch variability. Practical
two of the three batches should be at least pilot considerations prevent collection of broad
scale batches and the third one can be smaller amount of data because more data cause
7
, however US recommends at least 3 lots of more stress on analytical facilities and
product of a typical batch size should be tested increases the chances of errors. Testing
for stability8 whereas according to European fewer than three batches is allowed for stable
guidelines there are two options for selection of and well established products. In European
batches:

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guidelines for stability two options are available

Table 3.1 Pack details and Pack size

S. No. Pack No. of packs per Total No. of packs per

station Strength(mg) batch strength(mg)
25 50 100 25 50 100
1. PVC blister
(18tablets/blister) 40 30 25 640 480 400
2. TRIPLEX blister
( 18tablets/blister ) 40 30 25 640 480 400
3. SBP pack
1200 tablets = 25mg 2 2 2 6 6 6
600 tablets = 50mg
300 tablets =100mg
PVC – Polyvinyl chloride
SBP Pack – Simulated Bulk transfer Pack

information about different types of containers

for selection of batches and closures used in stability testing. The

For the conventional dosage forms and
when the active substances are known to be protocol must provide information about the
stable, stability data on at least two pilot type, size and sources of containers and
scale batches is acceptable Or closures. The purpose of stability testing is
not to qualify the container closure

For the critical dosage forms and when the
component labels, adhesives, colorants, inks,
active substances are known to be unstable,
etc. but to determine the expiry period for the
stability data on three primary batches is to
container closure combination as a whole.
be provided. Two of three batches should
European guidelines suggest to conduct
be of at least pilot scale and third batch may
testing on samples enclosed in containers
be smaller.
and closures proposed for storage and
As we are manufacturing sildenafil as a distribution. We selected three types of
conventional dosage form (tablets) and it is packaging material for Sildenafil tablets as
known to be a stable molecule, therefore we shown in Table 3.1. We selected 40, 30 and
have selected two batches of sildenafil of each 25 packs of PVC blister and TRIPLEX blister
strength, i.e., 25, 50 and 100 mg for conducting for strengths 25 mg, 50 mg and 100 mg
accelerated stability studies and the size of the respectively, required at each station and 2
batches for these different strengths are simulated bulk transfer (SBP) packs for all
200000 (tablets), 110000 (tablets), 105000 strengths. Total number of packs required per
(tablets) for 25 mg, 50 mg and 100 mg batch are 640, 480, 400 packs of PVC and
strengths respectively. TRIPLEX blister for strengths 25 mg, 50 mg,
100 mg and 6 SBP for all strengths as
required for 16 stations, i.e., one for initial
2. Containers and closures, Pack details
testing, four for accelerated testing ( 1, 2 , 3
and pack size
and 6 months), four for intermediate testing
The stability protocol must contain the
(3, 6, 9, 12 months) and seven for long term
testing (3, 6, 9, 12, 18, 24, 36 months).

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3. Sampling test time points 4. Storage conditions

According to European guidelines sampling Generally, a finished product should be
frequency of every 3 months during the first evaluated under storage conditions that test its
year, every 6 months during the second year thermal stability and if necessary its sensitivity
and then annually for drug products is to moisture and potential for solvent loss. As
suggested for long term testing whereas, per European guidelines storage conditions
testing frequency of 1, 2, 3 and 6 months is selected for sildenafil tablets are as shown in
recommended for accelerated stability testing. Table 3.2.
So we have selected 1, 2, 3 and 6 months for
accelerated testing of Sildenafil tablets.

Table 3.2 Storage conditions according to European guidelines

Study Storage condition Minimum time period
covered by data at
submission

Long term 25°C ± 2°C/60% RH ± 5% RH or 6 months (option a)

30°C ± 2°C/65% RH ± 5% RH 12 months (option b)

Intermediate 30°C ± 2°C/65% RH ± 5% RH 6 months

Accelerated 40°C ± 2°C/75% RH ± 5% RH months

Storage conditions for sildenafil tablets

Accelerated 40°C ± 2°C/75% RH ± 5% RH 6 months

5. Test parameters is quantitative (For example Moisture content,

The next step in stability protocol is to decide the Assay, degradation product, particle size,
test parameters. The parameters used for microbial growth, etc.) or it can be descriptive
evaluation of the drawn sample vary with type of if result is qualitative (For example Odour,
dosage form. In addition to drug assay and colour, appearance, cracking) that can be
analysis of the degradation product(s) one expressed as pass or fail. According to
should consider physical, chemical, biological European guidelines following tests are
and microbiological characteristics of the drug considered applicable to drug product
product(s) which are susceptible to change (Sildenafil tablets) and corresponding
during storage. The properties specific to a acceptance limits are also set by considering
particular dosage form should also be included in these guidelines.
the test parameter list, including dissolution of 6.1 Description
the solid dosage form, because it can vary as This parameter involves description of the
product age11.’ dosage form which is under consideration
6. Setting specifications and acceptance (e.g., size, shape, and color). If any of these
criteria characteristics change during manufacture or
For different test parameters included in the storage, this change should be investigated
stability study acceptance criteria should be fixed and corrective measure should be taken. The
before performing stability study. The criteria can acceptance criteria should include the final
be set in the form of numerical limits if the result acceptable appearance of the drug product.

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Table 3.3 Test parameters for sildenafil tablets with Acceptance limits

S.No. Test parameters Acceptance limits Evaluation

time points
(months)

1. Description Physical appearance should meet 0, 1, 2, 3, 6

Should meet specification

2. Identification Initial
requirements

Drug release should be NLT 80%

3. Dissolution time 0, 1, 2, 3, 6
in 45 min

4. Assay (95 -105 %) 0, 1, 2, 3, 6

Related substances
5. -unknown impurity NMT 0.2%w/w 0, 1, 2, 3, 6
-Total impurity NMT 0.4%w/w

6. Disintegration time NMT 15 min Initial

For 25 mg = 154 ± 6mg

7. Average weight For 50 mg = 309 ±12mg
Initial
For 100 mg = 618 ± 24mg

8. Content Uniformity Should meet requirements Initial

0, 1, 2, 3, 6
9. Moisture content NMT 6.0
months

Microbial limits
NMT 1000cfu/g
10. (TAMC)
NMT100cfu/g 0, 6 months
(TYMC)
Must be absent
E.coli (per 1.0g)
NLT = Not Less Than
NMT = Not More Than
TAMC = Total Aerobic Microbial Count.
TYMC = Total Yeast Mould Count.
cfu = colony forming unit

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6.2 Identification drug substance, method of manufacture, and

Identification testing should set up the identity the intended use of the drug product15.
of the drug substance present in the drug
product and should be able to distinguish 6.7 Dissolution
between compounds of closely related The specification for solid oral dosage forms
structures. Identity tests should be specific for normally includes a test to measure release
the drug substance, for e.g. infrared of drug substance from the drug product. By
spectroscopy. Identification only by a single establishing appropriate test conditions and
chromatographic procedure is not regarded as sampling procedures as official in
being specific. However, the use of two pharmacopoeia single-point measurements
chromatographic procedures where the are normally considered to be suitable for
separation is based on different principles or immediate-release dosage forms whereas
combination of tests into a single procedure multiple time point sampling should be
such as HPLC/UV, HPLC/MS or GC/MS is performed for extended-release dosage
generally acceptable. We have selected forms, and two-stage testing may be
HPLC/UV chromatographic procedures for appropriate for delayed-release dosage
identification test of sildenafil tablets. forms.
For immediate-release drug products where
6.3 Disintegration changes in dissolution rate have been
For rapidly dissolving (dissolution >80% in 15 established to significantly affect
minutes at pH 1.2, 4.0 and 6.8) products bioavailability, it is desirable to develop test
containing drugs which are highly soluble conditions which can distinguish batches with
throughout the physiological range, unacceptable bioavailability.
disintegration may be substituted for For extended-release drug products, in vitro /
dissolution. Disintegration testing is most in vivo correlation may be used to establish
suitable when a relationship to dissolution has acceptance criteria when human
been recognized or when disintegration is bioavailability data are available for
shown to be more selective than dissolution12. formulations exhibiting different release
6.5 Water content rates16.
A test for water content should be included 6.8 Impurities
when appropriate. The acceptance criteria may Organic and inorganic impurities (degradation
be justified with data on the effects of hydration products) and residual solvents are included
or water absorption on the drug product. In in this category.
some cases, a Loss on drying procedure may Organic impurities arising from degradation of
be considered adequate; however, a detection the drug substance and impurities that arise
procedure which is specific for water (e.g. Karl during the manufacturing process for the drug
Fischer titration) is preferred14. product should be monitored in the new drug
6.6 Microbial limits product. Acceptance limits should be
Microbial testing is considered as an attribute of acknowledged for individual specified
Good Manufacturing Practice as well as of degradation products, which may include
quality assurance. Acceptance criteria should both identified and unidentified degradation
be set for the total count of aerobic products as appropriate and total degradation
microorganisms, the total count of yeasts and products. Process impurities arise from the
moulds, and the absence of specific drug substance synthesis are normally
objectionable bacteria (e.g. controlled during drug substance testing, and
Staphylococcusbaureus, Escherichia coli, therefore not included in the total impurities
Salmonella, Pseudomonas aeruginosa). These limit. However, when a synthesis impurity is
should be determined by suitable procedures, also a degradation product, its level should
preferably using Pharmacopoeial procedures at be monitored and included in the total
a time point in manufacture which is justified by degradation product impurity limit.
data. The type of microbial test and acceptance
criteria should be based on the nature of the

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6.9 Assay (ANOVA) remains one of the most commonly

A specific stability-indicating assay to determine used methods of statistical analysis in the
strength (content) should be included for all statistical sciences18. Analysis of variance
drug products. In many cases it is possible to allows us to test the differences in the means
take up the same procedure (e.g., HPLC) for of several different groups or populations. In
both assay of the new drug substance and order to test the hypothesis, an F statistic is
quantitation of impurities. Results of content calculated which compares the variation
uniformity testing for new drug products can be among the groups with the variation within
used for quantitation of drug product strength, if the group19. By comparing the observed F
the methods used for content uniformity are value with the table value of F and then
also appropriate as assays17. analyzing the significant difference we test
Based on or after considering all these our hypothesis.
European regulations for stability testing we
have set acceptance criteria and specification CONCLUSION
for sildenafil tablets. Test parameters and
acceptance criteria selected for sildenafil Generally the success of a stability study
tablets is as given in Table 3.3. depends upon the sound stability protocol. A
7. Test methods successful stability protocol not only assures
For the test parameters that are included in the about our study but also reduces the chances
stability protocol, it is necessary to establish the of failure of study. When we developed the
test methodology. For the assay of the drug stability protocol for European market we
product it is advised to establish a stability observed that even after the harmonization of
indicating method, this method should be Stability Study guidelines there are some
validated for specificity, accuracy, precision and regional requirements which must be fulfilled
linearity in the range in which drug is expected in order to get the access to the market of
to fall during stability testing. Test procedures interest. Therefore regional requirements
from European Pharmacopoeia are generally should be emphasized during development of
followed for performing tests included in stability protocol. The purpose of considering
stability protocol. regional requirements during protocol
The next step after the stability study has development is that this helps for registration
completed is to analyze the data by applying for marketing in that region and this in turn
appropriate statistical test like Analysis of indirectly results in higher level of reliability on
variance (ANOVA). The analysis of variance the quality of the drug product.

REFERENCES
1. Carstensen, J.T and Rhodes, C.T. Drug 4. www.fda.gov accessed in 1997, Topic –
stability principal and practices. Chapter Guidance for industry
1.Introductory review.3rd edition revised and 5. Ali, J and Khar, R. K. A text book of
expanded. New York; Marcel Dekker, 2000 dosage form design. Chapter 4.
Stabilization of pharmaceutical products
2. Beaman, J. Designing a globally acceptable and stability testing, 1st Edn, Published by
registration stability protocol. Birla publications Pvt. Ltd. New Delhi:
Pharmaceutical technology. Mar 2007. 104, (2005).
6. Barron, M.D. Implementing successful
3. Carstensen, J.T and Rhodes, C.T. Drug stability testing operations.
stability principal and practices. Chapter Pharmaceutical technology Analytical
1.Introductory review.3rd edition revised and chemistry & testing, 2003; 14.
expanded. New York; Marcel Dekker, 2000 7. www.ich.org accessed in 2003. Topic -
stability testing of new drug substances
and products

www.ijpbs.net Pharmaceutics
P 33
ISSN 0975-6299 Vol.1/Issue-4/Oct-Dec.2010

8. www.fda.gov accessed in December 2008. healthcare council of Europe, Germany:

Topic – guidance for industry drug stability 2007, 6; 263-264.
guidelines 15. Pharmacopoeia European. Vol. I:
9. www.emea.eu.int accessed in 2000. Topic – Biological tests and general text on
stability testing of new drug substances and microbiology. Published by Directorate of
products. medicinal and healthcare council of
10. www.who.org accessed in 2009. Topic - Europe, Germany: 2007, 6; 529-531.
Stability testing of active pharmaceutical 16. Pharmacopoeia European. Vol.I:
ingredient and finished pharmaceutical Pharmaceutical technical procedures.
product. Published by Directorate of medicinal and
11. www.emea.eu.int accessed in 2003. Topic – healthcare council of Europe, Germany:
Guidance on stability testing: Stability 2007, 6; 266-274.
testing of existing active substances and 17. www.emea.eu.int accessed in 2006. Topic
related finished products. – Specifications: Test procedures and
12. Pharmacopoeia European. Vol. I: Assays. acceptance criteria for new drug
Published by Directorate of medicinal and substances and new drug products:
healthcare council of Europe, Germany: Chemical substances.
2007, 6; 278. 18. Steiger, J.H. Beyond the f-test: effect size
13. Pharmacopoeia European. Vol. I: confidence intervals and tests of close fit
Pharmaceutical technical procedures. in the analysis of variance and contrast
Published by Directorate of medicinal and analysis.
healthcare council of Europe, Germany: Physiological methods, 9(2): 164-182,
2007, 6; 278. (1995).
14. Pharmacopoeia European. Vol. I: Assays. 19. www.visualstats.org accessed in 2000.
Published by Directorate of medicinal and Topic – Analysis of variance.

www.ijpbs.net Pharmaceutics
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