Passage
Lyophilization, also known as freeze drying, is a method employed to eliminate moisture from samples. By utilizing sublimation, lyophilization directly converts frozen water, in its solid state, into water vapor. This approach presents advantages over liquid-based solvent removal techniques because it can be carried out at low temperatures, minimizing potential damage to samples that are sensitive to heat.
To perform lyophilization, a chamber containing the sample solution is connected to a lyophilizer. The lyophilizer first freezes the sample at a temperature below 0°C and then creates a vacuum, maintaining a pressure below 0.006 atm. Under these specific conditions, the frozen water undergoes sublimation, and the resulting water vapor is drawn from the sample chamber into a condenser set at -50°C. In the condenser, the water vapor is refrozen and held stationary.
The accompanying diagram depicts the phase diagram for water, illustrating the distinct negatively inclined equilibrium line between the solid and liquid phases. At low pressures, the solid phase of water is preferred, as long as any solute present in the water remains reasonably diluted. However, as the concentration of solute increases, the freezing point of water decreases. If the sample becomes concentrated enough, a decrease in pressure may cause it to melt rather than sublime. Furthermore, volatile solvents often cannot be eliminated from the chamber through the condenser, as their freezing point at low pressures falls below -50°C. The presence of volatile solvents in the gaseous environment can further contribute to impurities in the remaining water, thereby favoring the liquid phase even more.
Figure 1 Phase diagram of water
Question for Practice Passage Test - 3
Try yourself:The addition of which of the following substances is most likely to result in the melting of a sample during lyophilization?
Explanation
The passage states that volatile chemicals cannot be used in samples subjected to lyophilization because they will hinder sublimation and favor the liquid phase through melting. This is because these compounds have a high vapor pressure and cannot be removed from the system by the condenser. Choices A, B, and D are all hydrogen donors and acceptors. Choice C, diethyl ether, can only act as a hydrogen bond acceptor. Therefore, it has the weakest intermolecular forces and is the most volatile.
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Question for Practice Passage Test - 3
Try yourself:If the temperature inside the chamber is -60°C, what pressure must the chamber be at in order for the sublimation reaction to be at equilibrium?
Explanation
On a P vs. T diagram, sublimation equilibrium is indicated by the line separating the solid and vapor phases. On the graph in Figure 1, it is shown that at -60°C, the solid/gas line is at P = 0.0003 or 3 × 10-4 atm.
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Question for Practice Passage Test - 3
Try yourself:Lyophilization would most likely be employed industrially in which of the following separations?
Explanation
Both dichloromethane and methanol are removed as liquids at reasonably low temperatures and have freezing points far lower than water. This eliminates choices A and D. The passage states that one of the major advantages to lyophilization is that it can be performed at low temperatures, and therefore preserve the activity of heat-sensitive samples. NaCl is a salt that is stable at high temperatures, whereas enzymes are proteins that denature at high temperatures. Therefore, lyophilization is best suited as a means to remove water from aqueous protein solutions.
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Question for Practice Passage Test - 3
Try yourself:Sublimation of water inside the sample chamber causes the temperature of the chamber to:
Explanation
Sublimation is an endothermic reaction, requiring heat input. As this reaction removes heat from the surroundings, it lowers the temperature of the surroundings, in this case, the reaction chamber. This is very similar to sweat cooling the body as it evaporates off of the skin (also an endothermic process).
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Question for Practice Passage Test - 3
Try yourself:The heat of sublimation of water is 46 kJ/mol. If heat is transferred to the sample by the environment at a rate of 0.1 kJ/min, approximately how long will it take to lyophilize 40 cm3 of frozen water (density = 0.91 g/mL) ?
Explanation
40 cm3 of ice is 36 g or 2 moles of water. The heat required to sublimate this sample is 46 kJ/mol(2 mol) = 92 kJ. If heat is transferred at 0.1 kJ/min, then 920 minutes are required. Dividing 920 min by 60 min/hour gives just over 15 hours.
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