Biopharmaceutical products in storage change as they age, but they are considered to be stable as

long as their characteristics remain within the manufacturer's specifications. The number of days
that the product remains stable at the recommended storage conditions is referred to as the shelf
life. The experimental protocols commonly used for data collection that serve as the basis for
estimation of shelf life are called stability tests.
Shelf life is commonly estimated using two types of stability testing: real-time stability tests and
accelerated stability tests. In real-time stability testing, a product is stored at recommended
storage conditions and monitored until it fails the specification. In accelerated stability tests, a
product is stored at elevated stress conditions (such as temperature, humidity, and pH).
Degradation at the recommended storage conditions can be predicted using known relationships
between the acceleration factor and the degradation rate.
Temperature is the most common acceleration factor used for chemicals,
pharmaceuticals, and biological products because its relationship with the
degradation rate is characterized by the Arrhenius equation. Several methods
of predicting shelf life based on accelerated stability testing are described in
the article. Humidity and pH also have acceleration effects but, because they
Figure 1. A are complex, they will not be discussed in detail here. Also, details on
simulated set of statistical modeling and estimation are outside the scope of the article, but we
stability results provide references to computer routines.
Regulations and History The assessment of shelf life has evolved from
also showing examining the data and making an educated guess, through plotting, to the
the estimated application of rigorous physical-chemical laws and statistical techniques.
degradation and Regulators now insist that adequate stability testing be conducted to provide
95% confidence evidence of the performance of a drug or a biopharmaceutical product at
limits. different environmental conditions and to establish the recommended storage
conditions and shelf life.1-3 Recently, Tsong reviewed the latest approaches to
statistical modeling of stability tests,4and ICH has published some guidelines for advanced testing
design and data analysis.5,6
Modeling has become easier due to availability of standard statistical software
that can perform the calculations. However, an understanding of the general
principles of stability testing is necessary to apply these programs correctly and
obtain appropriate results. Thus, the purpose of this paper is to provide an
outline of the basic approaches to stability testing, as well as to create a
foundation for advanced statistical modeling and shelf life prediction. Table 1.
Stability and Degradation Since degradation is usually defined in terms of loss Estimates of the
of activity or performance, a product is considered to be degrading when any degradation
characteristic of interest (for example potency or performance) decreases. model and
Degradation usually follows a specific pattern depending on the kinetics of the Table 2.
chemical reaction. The degradation pattern can follow zero-, first-, and second- Estimates of
order reaction mechanisms.6 In zero-order reactions, degradation is independent degradation
of the concentration of remaining intact molecules; in first-order reactions, rates, days of
6,7
degradation is proportional to that concentration. Zero- and first-order stability and
reactions involve only one kind of molecule, and can be described with linear or 95% confidence
exponential relationships. Second- and higher-order reactions involve multiple limits.
interactions of two or more kinds of molecules and are characteristic of most
biological materials that consist of large and complex molecular structures. Although it is
common to approximate these reactions with an exponential relationship, sometimes their
degradation pattern needs to be modeled more precisely, and no shortcuts will suffice.
The degradation rate depends on the activation energy for the chemical reaction and is product
specific. We don't always have to deal with higher-order equations; in many cases, the observed
responses of different orders of reactions are indistinguishable for products that degrade slowly.
The degradation rate depends on the conditions where the chemical reaction takes place. Products
degrade faster when subjected to acceleration factors such as temperature, humidity, pH, and
radiation. Modeling of the degradation pattern and estimation of the degradation rate are
important for assessing shelf life. Experimental protocols used for data collection are called
stability tests. In practice, evaluators use both real-time stability tests and accelerated stability
tests. The real-time stability test is preferable to regulators. However, since it can take up to two
years to complete, the accelerated tests are often used as temporary measures to expedite drug
introduction.
Real-Time Stability Tests In real-time stability tests, a product is stored at recommended storage
conditions and monitored for a period of time (ttest). Product will degrade below its specification,
at some time, denoted ts, and we must also assure that ts is less than or equal to ttest. The estimated
value of tscan be obtained by modeling the degradation pattern. Good experimental design and
practices are needed to minimize the risk of biases and reduce the amount of random error during
data collection. Testing should be performed at time intervals that encompass the target shelf life
and must be continued for a period after the product degrades below specification. It is also
required that at least three lots of material be used in stability testing to capture lot-to-lot
variation, an important source of product variability.1,2