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Lyophilisation: difference between revisions

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Revision 12 — 04:03, 28 Dec 2024
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1{{Infobox method1{{Infobox method
2| name = Lyophilisation2| name = Lyophilisation
+3| subtitle = Freeze-drying
3| image = vial.svg4| image = vial.svg
4| caption = A lyophilised cake occupies approximately the volume of the original fill. Cake structure is a recorded quality attribute.5| caption = A lyophilised cake occupies approximately the volume of the original fill. Cake structure is a recorded quality attribute.
66Figures are collated from the freeze-drying literature and vary with concentration and with the measurement method.{{r|nail2002,tang2004}} The presence of sodium chloride is significant for peptide formulations because its low eutectic temperature drags the critical temperature of the whole system downward, forcing a colder and therefore longer primary drying step. A formulation carrying appreciable [[Trifluoroacetate counterion|trifluoroacetate]] or phosphate salt from purification behaves similarly.67Figures are collated from the freeze-drying literature and vary with concentration and with the measurement method.{{r|nail2002,tang2004}} The presence of sodium chloride is significant for peptide formulations because its low eutectic temperature drags the critical temperature of the whole system downward, forcing a colder and therefore longer primary drying step. A formulation carrying appreciable [[Trifluoroacetate counterion|trifluoroacetate]] or phosphate salt from purification behaves similarly.
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+69Annealing — holding the frozen product for a period above Tg′ but below the melting point — allows small ice crystals to grow at the expense of smaller ones and permits crystallising excipients such as mannitol to complete crystallisation before drying begins. Its benefit is a more uniform and less resistant dried structure; its cost is cycle time and, for some proteins, additional exposure to the ice-water interface.{{r|kasper2011}}
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68== Primary drying ==71== Primary drying ==
69Primary drying removes the ice, which for a typical aqueous fill is 90–98% of the water present. Its duration is set by the sublimation rate, which in the simplest treatment follows from an energy balance across the vial base: the mass flow equals the heat flow divided by the enthalpy of sublimation.72Primary drying removes the ice, which for a typical aqueous fill is 90–98% of the water present. Its duration is set by the sublimation rate, which in the simplest treatment follows from an energy balance across the vial base: the mass flow equals the heat flow divided by the enthalpy of sublimation.
77{{math|dm/dt = 0.126 W ÷ 2.83 × 10^{6} J kg^{-1} = 4.4 × 10^{-8} kg s^{-1} ≈ 0.16 g h^{-1}}}80{{math|dm/dt = 0.126 W ÷ 2.83 × 10^{6} J kg^{-1} = 4.4 × 10^{-8} kg s^{-1} ≈ 0.16 g h^{-1}}}
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+82A 2.0 mL fill therefore requires approximately 12.5 hours of primary drying under these conditions, before any safety margin is added. Doubling the shelf-to-product temperature difference halves that figure, which is why the choice of critical temperature — and how close the cycle is permitted to approach it — dominates cycle economics.{{r|tang2004,rambhatla2003}}
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79=== Collapse and eutectic melting ===84=== Collapse and eutectic melting ===
80Two distinct structural failures bound primary drying. In a system whose freeze-concentrate crystallises, exceeding the eutectic melting temperature produces liquid, and the vial dries as a shrunken residue or a glassy shell — usually described on inspection as melt-back. In an amorphous system, exceeding a temperature a few degrees above Tg′ allows viscous flow of the freeze-concentrated matrix into the pores that the sublimed ice has vacated. The dried layer loses its structure, and the result is a shrunken, sometimes glassy or foamed cake. The temperature at which this becomes observable by freeze-drying microscopy is the collapse temperature, Tc, conventionally 1–3 °C above Tg′ for small-molecule and peptide systems.{{r|tang2004}}85Two distinct structural failures bound primary drying. In a system whose freeze-concentrate crystallises, exceeding the eutectic melting temperature produces liquid, and the vial dries as a shrunken residue or a glassy shell — usually described on inspection as melt-back. In an amorphous system, exceeding a temperature a few degrees above Tg′ allows viscous flow of the freeze-concentrated matrix into the pores that the sublimed ice has vacated. The dried layer loses its structure, and the result is a shrunken, sometimes glassy or foamed cake. The temperature at which this becomes observable by freeze-drying microscopy is the collapse temperature, Tc, conventionally 1–3 °C above Tg′ for small-molecule and peptide systems.{{r|tang2004}}
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82Collapse is not necessarily a chemical failure. A collapsed cake may reconstitute and may assay within specification. Its documented consequences are elevated residual moisture, because the collapsed structure traps water; slower and sometimes incomplete reconstitution; and a cake appearance that fails a visual specification. For proteins, collapse has been associated with reduced storage stability, attributed to increased molecular mobility in the denser matrix.{{r|wang2000}}87Collapse is not necessarily a chemical failure. A collapsed cake may reconstitute and may assay within specification. Its documented consequences are elevated residual moisture, because the collapsed structure traps water; slower and sometimes incomplete reconstitution; and a cake appearance that fails a visual specification. For proteins, collapse has been associated with reduced storage stability, attributed to increased molecular mobility in the denser matrix.{{r|wang2000}}
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+89Conservative practice therefore sets the target product temperature 2–5 °C below Tc and accepts the longer cycle. Aggressive cycles that run within a degree of Tc exist and are defensible when the critical temperature has been measured for the specific formulation rather than assumed from a table.{{r|tang2004}}
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84== References ==91== References ==