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LYOPHILIZATION

Lyophilization: Heat and Mass Transfer

Narlin Beaty, Ph.D. Sublimation Science

Abstract

n the lyophilization of pharmaceuticals, the product sublimation interface temperature must be kept below the product collapse temperature to achieve pharmaceutical elegance and assure stability. Currently, meaningful equipment controls are only available for chamber pressure and shelf temperature. This review derives and explains the use of the heat and mass transfer equation for predicting these control parameters in a manner that meets the interface temperature condition. The use of this method has substantially reduced the amount of trial and error associated with lyophilization cycle development.

to avoid collapse, the interstitial fluid must also be solidified (probably as a glass). Thus, a goal is to conduct primary drying at a temperature and pressure that will keep all of the components in a solid state. Many combinations of ingredients will not crystallize (hence the lack of a eutectic) and instead solidify as glass at relatively low temperature. For example, sucrose, salt, and water form a glass that solidifies in a range around -40oC. Such excipient combinations can still be sublimed, but under conditions which maintain the product below the aforementioned glass transition temperature.

Surface Collapse
Pharmaceutical lyophilization is all about the phase change of water from solid to gas as a consequence of heat energy supplied through the shelves. Were water the only concern, sublimation could be conducted at any temperature below its freezing point. Knowing a products collapse temperature, the sublimation of water-ice can be performed slightly lower than the product collapse temperature and maintain the frozen structure. Only surface temperatures matter. Internally, structure is maintained as long as crystallized molecules are present, be they water-ice, mannitol, or other excipients. If temperature at the side surface of a vial rises above the collapse temperature, sublimation will cause channels along the sides and a loose cake. The channels can go to the bottom of the vial and cause partial collapse along the relatively warmer bottom surface.

Introduction
Starting as early as 1993, FDA in the Guide to Inspections of Lyophilization of Parenterals established that, Obviously, the manufacturer should know the eutectic point . . . [1]. Yet it has remained not obvious to many manufacturers why the information is needed or what value it adds. Since then the industry has clarified the eutectic terminology to include either the collapse temperature or the glass transition temperature, Tg, as an acceptable alternative because many products do not crystallize and do not have a classical eutectic point. That same year, Kochs et al. [2] showed by example how the collapse temperature data could be used to maximize the sublimation rate. Other authors have repeated the observation and perhaps some have even used the collapse temperature data in calculations to obtain set points for shelf temperature and chamber pressure. Most users have, at best, used the original graph of Kochs or a copy produced elsewhere [3]. The purpose of this review is to explain how to use the collapse temperature with simple equations to predict appropriate settings for the shelf temperature and chamber pressure during primary drying.

Ohms Law, Clausius Clapeyron, and the Rate of Sublimation

Ohms Law states that voltage is equal to the product of amperage and resistance. The analogous relationship in fluids is Pressure = Flow x Resistance. Flow is the change in mass with time, m/t and concentration is measured by gas pressure, hence m/t=P / Resistance. Where is the change in pressure, P , to be observed? Where is the Resistance? Clausius Clapeyron taught that a sublimating ice would maintain a specific temperature and pressure relationship.
Hsub R Ti

Goal
Primary stage collapse is the result of ice crystal sublimation at a temperature where the product-water solution is not completely frozen. It can occur in some or the entire product and the result is shrunken cakes, crusty tops or bottoms, disappearing product, discoloration, and other similar descriptions. When water-ice crystallizes, all non-water components of the product and some of the water are concentrated between the ice crystals resulting in a low solidification temperature for the interstitial fluid. So while ice may be maintaining macroscopic structure at temperatures as high as 0oC, when primary drying proceeds and the ice sublimes, unsolidified product constituents will settle (collapse) onto the sublimation surface. In order 1

P (Ti)

[EQ 1: Clausius Clapeyron]

A e

Where Hsub = 51027 J/mol, R = 8.314772 J/mol K, and A = 3.493 1012 Pa. The pressure at the interface, P(Ti), is a function of the interface temperature. Lyophilizers control the chamber pressure, but as sublimation proceeds, gas phase water molecules near the sublimation interface reflect from dried product and cause a pressure differ

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LYOPHILIZATION
ential between the chamber and the sublimation front. That tiny difference in pressure permits a calculable increase in interface temperature, with resistance increasing as the product dried layer grows. Additionally, albeit less significant, resistance is caused by the small openings between the stopper and vial and even by the walls of the vial itself. Letting Pi and Pc represent pressure at the interface and chamber respectively, and Ap and Rp the product area and resistance one can write a lyophilization rate, where the units of Rp are Pam2skg-1 which will unit reduce to m/s. The units of rate will be kg/s (SI units) or gm/hr (conventional).

Figure 1.

A p

(P i Pc)
Rp

[EQ 2: Rate Equation from Ohms Law]

It is common in lyophilization to express rates in area normalized units, dividing through by Ap. The conversion from Rp in m/s to Rp in cm2torrhrgm-1 is Rp(m/s) x 2.08310-5.

Heat Transfer is from the shelf set point through the air space and the glass to the bottom of the ice. Heat transfer then continues through the ice from bottom to top. The top of the ice is called the interface. As sublimation proceeds, the interface moves toward the bottom of the vial, leaving a dried product layer behind.

Now using the rate equation from Ohms Law (EQ 2), we can substitute for the m/t term to get

Newtons Law of Cooling

Newtons law of cooling says the rate at which an object gains or loses heat is proportional to the difference between its temperature and the ambient temperature, Temp/t=k (Ambient-Temp). For the purpose of lyophilization, the proportionality constant, k, is an overall heat transfer coefficient, Kv, times the vial bottom area, Av. The heat transfer coefficient contains energy units, J/m2 s K, where K is Kelvin temperature. Conventionally, the change in temperature is related to a change in heat, Q/t.

A v K v Ts Ti T

Ap P i P c Hsub Rp Mw

This can be solved for Pi, pressure at the sublimation interface, and the Clausius Clapeyron expression (EQ 1) can be substituted in place of Pi. One then arrives at the following useful expression.
Hsub RTi

A v K v Ts T b

Ae

A p Hsub Pc + Av Kv Rp M w T s T i T Ap Hsub

[EQ 3: Newtons Law of Cooling]

[EQ 5: Coupled Heat & Mass Transfer Equation] Equation 5 can be numerically solved for Ti and values of Ts and Pc can be used to predict a product ice interface value for comparison to the measured collapse temperature. If Ti is too much less than the collapse temperature, then the sublimation rate will be slower than need be. If it is close to, or greater than the collapse temperature, then collapse will occur. Alternatively, Equation 5 can be solved for chamber pressure, Pc. If the optimum cycle calls for Ti to be 2OC less than the collapse temperature, one can simply calculate a chamber pressure using Ti and a range of selected shelf temperatures.

The constant temperature source (ambient temperature) is the lyophilizer shelf, Ts, while Tb is the temperature on the inside bottom of the vial. However, the temperature of interest is at the top of the ice. Although the temperature change, T, between the bottom and top of the ice, varies from one product to the next, it is about 1.6OC/cm in the range of -30OC to -40OC.

Thermodynamics of Open Systems

From thermodynamics another expression is commonly used for the time change of heat.

Hsub
Mw

[EQ 4: Thermodynamics]

Mw

0.018

kg mol

Resistance: Rp
An average value can be assumed for Rp. In reality, resistance changes with time and, in any event, is never exactly zero. When lyophilizing water from a vial, even without a stopper, the vial walls limit degrees of freedom and offer some resistance to flow such that the interface pressure is greater than the chamber pressure. The selection of a resistance value has been extensively investigated by Rambhatla et al.[4]. Product resistance from the dried cake varies based on the percentage of solids, the molecular entities used, and the temperature at which nucleation for freezing occurs. The later is the least controlled, but is more uniform after a short (30 minute to 1 hour) annealing step, where the ice temperature is held slightly above the collapse temperature. Existing product lyophilization data can be used to solve for an estimate of Rp. Since the product thermocouple temperature, Tb (at the bottom of the ice), is close to the product interface temperature (top of the ice), one can use Tb and EQ 2 to solve for Pi, and then a rate equation to obtain Rp, m/t=(Pi-Pc)/Rp. A better estimate of Rp is obtained from the manometric temperature method (MTM)5, but that data is less commonly available. 2

The units of Q/t are energy/time, J/hr. Q/t is just the change in mass times the energy inside the system, which for lyophilization, is the enthalpy of ice sublimation.

Assembly
Using this basic understanding from multiple areas of science, we have assembled four equations which can be grouped to yield a rather spectacular result. Equating the heat change from Newtons Law of Cooling (EQ 3) and the thermodynamics of open systems (EQ 4), we get the following.

Av Kv T s T i T

Hsub
Mw

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LYOPHILIZATION Heat Transfer Coefficient: Kv

The heat transfer coefficient, Kv, is not obtained from a table for glass. Because the vial touches the shelf at only a few places and varies in thickness across its bottom, there is significant transfer of heat from the shelf to air and from air to the vial. Consequently, a change in chamber pressure causes a change in Kv. The following equations and Figure 2 have been used to estimate Kv. each time letting the sublimation come to equilibrium as determined by stable product thermocouple temperatures. For nearly all vials and even for syringes, with no glass to shelf contact, values range from about 5 to 33 J/m2 s K (1.2 10-4 to 7.9 10-4 cal cm-2 s-1 K-1). Clearly with syringes, the meaning of the Av term is not obvious, but can be used as the outer diameter of the syringe. In order to benefit from Equation 5, it is necessary to estimate and use Kv as a function of chamber pressure. Another way to estimate Kv at any one pressure has been suggested by Pikal [6]. One can measure the rate of sublimation in units of kg/(m2 s) and apply the following formula.

J 51.981 KC = + 5.872 3 2 vial cm s m K

s
2 2

A p Rate
vial + 0.025 s
2

KD

250

m s kg

if vialtype
2

"tubing"

Kv

kg m

Av T s T b

Hsub Mw

Calculation of Kv at a single chamber pressure.

m s 191.429 vial + 0.033 2 kg kg m

if vialtype

"molded"

These equations and Figure 2 were derived from the data of Tang and Pikal [6]. The shape of these curves is correct, but examples exist where a calculated Kv at one pressure was lower than expectation. In that case, and where it is not possible to repeat work at various pressures, it is reasonable to divide the Kv(Pc) expression by a factor necessary to reduce Kv(Pc) to the value actually measured and thereby preserve the pressure dependence. Low values of Kv will translate into slower sublimation rates for any given shelf temperature. Care must be exercised in choosing a function for Kv(Pc). The formulas presented here are from a limited data set and in any event would not be specific for an arbitrary vial. Indeed, some lyophilizations are conducted from syringes and ampoules. A calculation of Kv for the container and any one pressure can be made from Equation 5.
Hsub

Still, the rate is best derived from data obtained at multiple pressures and Kv(Pc) should be expressed as a fitted function of chamber pressure.

Cycle Analysis
Using EQ 5 and a function Kv(Pc), the classic graph of shelf temperature, chamber pressure, and product interface temperature versus rate can be constructed, unique to a product, vial, cycle, and lyophilizer combination. Two solutions to Equation 5 are shown. In the first case, it is solved for Ti as a function of shelf temperature and chamber pressure, and in the second case it is solved for Pc as a function of interface temperature and shelf temperature.

Case 1:
f P c ,Kv ,Ti ,Ts

K v Pc

( )

Ap H sub

Pc + A e

R T

A p Hsub Pc + Av Kv Rp Mw Ts Ti T Ap Hsub

H sub

A e

R T

EQ 5 in homogeneous form

Av Rp Mw Tb Ts

Ti

A v Kv Pc Rp

( )

Mw A p Hsub

Hsub
R T i
2

H sub R Ti

Partial derivative of EQ 5

Using data from the steady state area of primary drying, the thermocouple temperature from a data set can be assumed to be Tb =

T i (TS , PC)

Tint 0 273.16K 5K for i 0 .. 9 f (Tinti ,TS , PC)

Figure 2. Dependence of the Heat Transfer Coefficient, Kv, on Pressure. These data were derived for four different vials and are not representative of all vials. Case 2:

Tint i+1 Tinti

f Ti

Iterative Solution for Ti.

Tint 9

From EQ 5 and the function Kv(Pc), one can solve for chamber pressure with varying shelf temperatures, obtaining a rearranged version of EQ 5, Pc(Ti,Ts)=function. A symbolic solution may be difficult depending on the complexity of Kv(Pc). For example:
g P c , Kv

m 1.043 P s K c KC + 1 + KD P const Kv [Homogeneous expression for c Kv(Pc).]

Ti + 1.6O (height of ice), and replaces Ti + T. To obtain Kv as a function of Pc, one must vary the chamber pressure widely and obtain thermocouple data at three or more points,

The const term is a scaling factor to preserve the curve shape while adjusting it to fit data from one pressure. With the two equations f(Pc,Kv) and g(Pc,Kv), one can use a Newton-Raphson solution for Pc as follows.

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f (P c , Kv ) P c Jacobian (Pc ,K v ) = g P ,K P ( c v ) c
Pc T i , T s :=

K v

g P ,K ( c v) K v

f P c , Kv

Figure 3.

"Start with any reasonable values for Pc and Kv" n0 Pc 20Pa

n

kg Kv 5 3 n s K for n 1 .. 4

Pc Pc n+ 1 n Jacobian P , K cn v n Kv Kv n+1 n

f Pc , K v , T i , T s n n n n g Pc , Kv n n

return Pc 4

Then the chamber pressure and shelf temperature can be used to calculate a rate, where Pi(Ti) is the Clausius Clapeyron function.
Rate T i ,T s

Pi T i Pc T i , Ts Rp

()

)
[EQ 6 Sublimation Rate at given Ti and Ts]

The rates, graphed as smooth curves in Figure 3, are derived from varying chamber pressure versus a solution to Equation 5 for Ti(Ts,Pc) as in Equation 7. Whereas the straight lines in Figure 3 are derived from Equation 6, with known values of Ti and Ts.
Rate1 Ts , Pc

Rate, Pressure, Temperature diagram for a specific product and vial. The curves represent projected rates versus chamber pressure for different shelf temperatures. The diagonal lines represent a constant interface temperature. The product was in a 20cc tubing vial with an estimated Rp equal to 16559 Pam2s/kg. The Kv pressure relationship followed the form shown in g(Pc,Kv) with const = 1/1.826, where limited data permitted Kv calculation at only one pressure. The crossing orthogonal lines with rate = 0.048 and pressure = 210mTorr represent an actual run condition. Axes have been translated to conventional units.

Pi T i Ts , Pc Rp

( (

)) Pc [EQ 7 Sublimation Rate given Ts and Pc]

Figure 4.

It is obvious that the maximum rate is obtained from a minimum pressure and a maximum shelf temperature. Consequently, it makes sense to look at a graph of Ti versus shelf temperature at the lowest chamber pressure that can be reliably maintained. For many older lyophilizers, that chamber pressure is about 50 mTorr.

Conclusion
Optimal sublimation rate and avoidance of product collapse is a goal in pharmaceutical lyophilization. We have presented the derivation and solution of those equations which can be used to find a suitable shelf temperature and chamber pressure after knowing a product collapse temperature. Although some research investigators have been performing these calculations for several years, they have not been easily embraced by most manufacturing departments, probably because the algebraic solutions typically utilize iterative methods. This paper has shown both a derivation for the central coupled heat and mass transfer equation, as well as methods for its solution. Other solutions and considerable complexity can be introduced. This method is not intended to represent a complete simulation of the lyophilization process. It is nothing more than an analysis of the physical primary drying parameters. A major consideration for the use of this analysis is that a vial-package heat transfer coefficient must be obtained as a function of pressure. Such work can either be done experimentally or by estimate. Also, the dry product layer resistance is treated as a single average number, when in fact it is known to continuously increase throughout the sublimation. Still, the method is greatly superior to having no analysis and no understanding of existing product lyophilization cycles.

Interface Temperature versus Shelf Temperature with chamber pressure at 50 mTorr. The product has a collapse temperature measured at -28OC and the chosen lyophilizer can reliably hold a 50 mTorr vacuum. This analysis suggests that for an interface temperature of -30OC, a shelf temperature of 34OC will maximize both the interface temperature and the sublimation rate at this pressure. www.fda.gov/ora/inspect_ref/igs/lyophi.html. 2. Kochs, M., Korber, C., Heschel, L., Nunner, B., 1993, The Influence of the Freezing Process on Vapour Transport During Sublimation in Vacuum-Freeze-Drying of Macroscopic Samples. Int. J. Heat Mass Transfer, 36, 1727-1738. 3. Franks, F., 1997, Freeze-Drying of Bioproducts: Putting Principles Into Practice. European Journal of Pharmaceutics and Biopharmaceutics, (45) 221-229. 4. Rambhatla, S., Ramot, R, Bhugra, C. Pikal, M.J., 2004, Heat and Mass Scale-up issues during Freeze Drying: II Control and characterization of the Degree of Supercooling. AAPS Pharm. Sci. Tech. 5(4) Article 58. 5. Milton, N., Pikal, M.J., Roy, M.L., Nail, S.L., 1997, Evaluation of 4

References
1. U.S. Food and Drug Administration, 1993, Guide to Inspections of Lyophilization of Parenterals. Office of Regulatory Affairs.

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Manometric Temperature Measurement as a Method of Monitoring Product Temperature During Lyophilization. PDA J. Pharm. Sci. Technology, (5) 7-16. 6. Pikal, M.J., Cardon, S., Bhugra, C., Jameel, F., Rambhatla, S. 2005, The Nonsteady State Modeling of Freeze Drying: In Process Product Temperature and Moisture Content Mapping and Pharmaceutical Product Quality Applications. Pharmaceutical Development and Technology, (1) 17-32.

Narlin Beaty, Ph.D. is a principal in Sublimation Science, a service organization to the parenteral pharmaceutical industry specializing in lyophilization commissioning, cycle development, and both machine and product cycle validation. He is also a founder of Qualification Process Solutions, an engineering firm that performs large equipment and utility commissioning, as well as managing production shutdown activities. His Ph.D. is from the Univ. of Michigan. To correspond with the author, please contact the editor at: nc@russpub.com

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