Effect of formulation on

lyophilization, part 1
Formulation components--their freezing
and drying
Thomas A. Jennings

The profitability of a lyophilized product begins with the

development of the formulation, not at the start of
manufacturing.
NOTE: This is the first part of a two-part article. Part 2 is also available for on-line
viewing.

A formulation can be defined as a liquid medium in which one or more active

components (chemical or biological) remain in a stable environment and maintain
specified potency limits for some period of time. For example, a vaccine formulation
may still be effective and safe to use after several days of storage at 4°C. After about
a week of storage, it must be discarded. However, if the kinetic clock for degradation
of the vaccine could be slowed so that 1 second was extended to 1 hour, then the
useful life of the formulation would be increased to almost 20 years.

Lyophilization stabilizes the formulation by slowing the kinetic clock of the

degradation process. It alters the clock by removing the solvent component or
components to levels that no longer support chemical reactions or biological growth.
This removal is accomplished, first, by freezing the formulation, that is, separating
the solutes from the solvent or solvents and immobilizing any solvent in the
interstitial region between the solvent crystals. Then the solvent is removed by
sublimation (primary drying) and next by desorption (secondary drying).

The nature of the formulation determines:

• Time and temperature needed to obtain a completely frozen matrix.

• Time, shelf temperature, and chamber pressure to conduct primary and
secondary drying.
• The nature of the container system.
• Design and construction of the freeze-drying equipment.

Because time is equated to productivity, the formulation affects not only the process
and drying equipment, but also the cost of manufacturing. As process time increases,
profitability decreases.

Components of a Formulation
 The formulation consists of three basic components--active ingredient,
excipient, and solvent system. In general, the active ingredient in the
pharmaceutical industry is defined by its potency and, in the
diagnostic industry, by its reactivity. Depending on means of
production, there may be variations in the composition of the active
component from batch to batch.

The formulation of a product affects the time

required to process it and, ultimately, the
cost of the product. Here, products are
readied for lyophilization at Medicus
Technologies (West Chester, PA).

Excipients serve several functions.1,2 They

primarily provide a stable liquid environment
for the active ingredient for some finite time.
The excipient may also cryoprotect the
active ingredient during the freezing
process.35 In the freezing of formulations
containing biological organisms, the
formation of ice within can lead to the
organism's destruction by cell membrane
rupture. Sucrose, glucose, and dextran are
excipients used to cryoprotect organisms.2,4

The excipient may also serve only as a bulking agent.2 When solid concentrations of a
formulation reach <2%, the resulting cake may have poor structural qualities and
leave the container during the drying process. The addition of bulking agents such as
mannitol and dextran strengthen cake structure. The role of the solvent system is
often overlooked. Most formulations are totally aqueous solutions, although others
contain solvents such as tertiary butyl alcohol to increase the solubility of some
compounds. The solvent system is removed during drying, but its thermal properties
have a major impact on the cosmetic properties of the final product (Figure 1).

Figure 1. Typical lyophilization cakes. (A) uniform distribution of constituents; (B)

nonuniform distribution of constituents in the cake with a crust or glaze on the upper
surface; (C) a cake with poor self-supporting structural properties; (D) a cake
showing signs of collapse; (E) example of meltback; (F) disappearing cake, i.e.,
dissolution of the cake by excess water; and (G) puffing resulting from incomplete
freezing of the matrix before evacuation of the dryer.

Freezing and Drying the Formulation

Freezing. Formation of ice during freezing results in dramatic changes in

concentrations of the active ingredient and the excipient or excipients of the
formulation. As an example, consider the freezing of an isotonic (0.9% w/v) NaCl
aqueous solution. At a temperature of 20°C, the frozen matrix consists of ice crystals
interlaced with a 23% NaCl solution. At a temperature of <23°C, the interstitial
 region in the matrix consists of a eutectic mixture of NaCl * 2H2O and ice
crystals. (Eutectic temperature is a point on a phase diagram where the
temperature of the system or the concentration of the solution at the
point cannot be altered without changing the number of phases present.)

In most formulations (>99%; based on thermal analysis of >1000

formulations at Phase Technologies, Inc., Conshohocken, PA) containing
an active ingredient and an excipient, freezing greatly increases the
concentration of the active ingredient and the excipient or excipients, but
does not produce a well defined eutectic mixture. Instead, freezing produces a
complex, glassy system that may be homogeneous or heterogeneous. This complex
system, at this time, can only be produced in the interstitial region of ice crystals as
a result of the freezing process.

Another property of frozen matrix is the degree of crystallization (D), the ratio of the
quantity of ice formed during the freezing process to the total freezable water in the
formulation (Jennings TA, "Lyophilization Seminar" [Notes], Conshohocken, PA, Phase
Technologies, Inc., p 26, 1994). As D approaches 1 (Figure 2), most water is in the
form of ice crystals, and only a small quantity forms part of the interstitial region.
The ice crystals interconnect to form vapor paths. With decreasing values of D (e.g.,
D = 0.5), the volume of glassy interstitial region approaches that of the ice crystals.

Figure 2. Illustration of a frozen matrix of a formulation in which the degree of

crystallization approaches 1.

In the frozen state, the mobility of the water in the glassy interstitial region
approaches 0, and the formulation is considered completely frozen. As the
temperature of the matrix increases, the glassy interstitial region softens, the
electrical resistivity of the interstitial region decreases, or the conductivity of the
system increases. Such a change in the electrical nature of the matrix is associated
with the onset of mobile water within its interstitial region. As temperature further
increases, the interstitial region slowly takes on liquidlike characteristics, while
surrounding ice crystals remain frozen.

In most glassy systems, onset temperature for the mobility of the water in the
interstitial region is not as sharp and well defined as that for a eutectic mixture. The
onset temperature for the mobility of water in the matrix interstitial region is referred
to as collapse temperature. This definition differs from that of MacKenzie, who called
collapse temperature "disappearance of the freezing pattern, more or less extensive
flow of the residual material, and the generation of new patterns."6

Drying. For lyophilization to occur, the solvent is first removed by sublimation while
the temperature of the frozen matrix is maintained below the eutectic or collapse
temperature of the formulation. This is the primary drying process. The chamber
pressure and product and shelf temperatures, during primary drying, are based on
the formulation's eutectic or collapse temperature. A lyophilized cake is typified by
Figures 1A and 1B. The resulting cake volume approaches the original fill-volume.

Primary drying at temperatures greater than that of the collapse or eutectic

temperature of the formulation (sometimes referred to as vacuum drying or
cryodrying) can lead to a product typified by Figures 1D and 1E, respectively. Figure
1D illustrates some collapse of the cake resulting from the presence of mobile water
in the matrix interstitial region during primary drying. Figure 1E, though, illustrates
meltback, a result of liquid states in the interstitial region.

After primary drying, the residual moisture on the resulting cake surface is reduced
to levels that no longer support biological growth and chemical reaction. This process
is secondary drying. The reduction of moisture in the cake during secondary drying is
accomplished by increasing the shelf temperature and reducing the partial pressure
of water vapor in the container. The required partial pressure of water vapor and
shelf temperature are ascertained from stability studies of lyophilized or vacuum-
dried products having varied amounts of residual moisture.

References

1. Bashir J, and Avis KE, "Evaluation of Excipients in Freeze-Dried Products for

Injection," Bull Parent Drug Assoc, 27:6883, 1973.

2. Wang YJ, and Kowal RR, "Review of Excipients and PH's for Parenteral Products
Used in the United States," Bull Parent Drug Assoc, 34:452462, 1980.

3. Greaves RIN, "Fundamental Aspects of Freeze-drying Bacterial and Living Cells," in

Aspects Théoriques et Industriels de la Lyophilisation, Rey L (ed), Paris, Herman, pp
407410, 1964.

4. Smith AU, "Some Problems in Supercooling and Freezing Living Cells, Tissues and
Organisms," in Aspects Théoriques et Industriels de la Lyophilisation, Rey L (ed),
Paris, Herman, pp 257278, 1964.

5. MacKenzie AP, "Comparative Studies on Freeze-drying Survival of Various Bacteria,

Gram Type, Suspending Medium and Freezing Rates," in International Symposium on
Freeze-drying of Biological Products, Washington, DC, Develop. Biol. Standard, 36 S,
Basel, Switzerland, Karger, pp 263277, 1977.

6. MacKenzie AP, "Collapse during Freeze-drying--Qualitative and Quantitative

Aspects" in Freeze Drying and Advanced Food Technology, Goldblith SA, Rey L, and
Rothmayr WW (eds), New York, Academic Press, pp 277306, 1975.

Thomas A. Jennings, PhD, is CEO, Phase Technologies, Inc.

(Conshohocken, PA).

Continue on to Part 2 of this article.

Copyright©1997 IVD Technology

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