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
Figure 1. A article. Humidity and pH also have acceleration effects but,
simulated because they are complex, they will not be discussed in detail
set of here. Also, details on statistical modeling and estimation are
outside the scope of the article, but we provide references to
stability
computer routines.
results also Regulations and History The assessment of shelf life has
showing the evolved from examining the data and making an educated
estimated guess, through plotting, to the application of rigorous physical-
degradation chemical laws and statistical techniques. Regulators now insist
and 95% that adequate stability testing be conducted to
provide evidence of the performance of a drug
confidence
or a biopharmaceutical product at different
limits. environmental conditions and to establish the
recommended storage conditions and shelf life.1-3 Recently,
Tsong reviewed the latest approaches to statistical modeling Table 1.
of stability tests,4and ICH has published some guidelines for
advanced testing design and data analysis. 5,6 Estimates of
Modeling has become easier due to availability of standard the
statistical software that can perform the calculations. degradation
However, an understanding of the general principles of model and
stability testing is necessary to apply these programs Table 2.
correctly and obtain appropriate results. Thus, the purpose of
Estimates of
this paper is to provide an outline of the basic approaches to
stability testing, as well as to create a foundation for degradation
advanced statistical modeling and shelf life prediction. rates, days
Stability and Degradation Since degradation is usually defined of stability
in terms of loss of activity or performance, a product is and 95%
considered to be degrading when any characteristic of confidence
interest (for example potency or performance) decreases. limits.
Degradation usually follows a specific pattern depending on the kinetics of
the chemical reaction. The degradation pattern can follow zero-, first-, and
second-order reaction mechanisms.6 In zero-order reactions, degradation is
independent of the concentration of remaining intact molecules; in first-order
reactions, degradation is proportional to that concentration. 6,7 Zero- and first-
order reactions involve only one kind of molecule, and can be described with
linear or exponential relationships. Second- and higher-order reactions
involve multiple 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 (t test).
Product will degrade below its specification, at some time, denoted t s, 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