Evaluation of manometric temperature measurement, a process analytical technology tool for freeze-drying: Part I, product temperature measurement

This study examines the factors that may cause systematic errors in the manometric temperature measurement (MTM) procedure used to evaluate product temperature during primary drying. MTM was conducted during primary drying using different vial loads, and the MTM product temperatures were compared with temperatures directly measured by thermocouples. To clarify the impact of freeze-drying load on MTM product temperatures, simulation of the MTM vapor pressure rise was performed, and the results were compared with the experimental results. The effect of product temperature heterogeneity in MTM product temperature determination was investigated by comparing the MTM product temperatures with directly measured thermocouple product temperatures in systems differing in temperature heterogeneity. Both the simulated and experimental results showed that at least 50 vials (5 mL) were needed to give sufficiently rapid pressure rise during the MTM data collection period (25 seconds) in the freeze dryer, to allow accurate determination of the product temperature. The product temperature is location dependent, with higher temperature for vials on the edge of the array and lower temperature for the vials in the center of the array. The product temperature heterogeneity is also dependent upon the freeze-drying conditions. In product temperature heterogeneous systems, MTM measures a temperature close to the coldest product temperature, even, if only a small fraction of the samples have the coldest product temperature. The MTM method is valid even at very low product temperature (−45°C). Published: February 10, 2006     

Effect of potassium citrate salts on the vapor-liquid equilibrium properties of aqueous solutions of alanine at different temperatures

Water activity measurements by isopiestic method have been carried out on the aqueous solutions of alanine + potassium di-hydrogen citrate (KH₂Cit) and alanine + tri-potassium citrate (K₃Cit) over a range of temperatures at atmospheric pressure. From these measurements, values of the vapor pressure of solutions were determined. The effect of temperature and charge on the anion of salts on the vapor–liquid equilibrium of the investigated systems has been studied. The experimental water activities have been correlated successfully with the segment-based local composition Wilson and NRTL models. The agreement between the correlations and the experimental data is good.     

Moderate temperatures affect Escherichia coli inactivation by high-pressure homogenization only through fluid viscosity

The inactivation of suspensions of Escherichia coli MG1655 by high-pressure homogenization was studied over a wide range of pressures (100-300 MPa) and initial temperatures of the samples (5-50 degrees C). Bacterial inactivation was positively correlated with the applied pressure and with the initial temperature. When samples were adjusted to different concentrations of poly(ethylene glycol) to have the same viscosity at different temperatures below 45 degrees C and then homogenized at these temperatures, no difference in inactivation was observed. These observations strongly suggest, for the first time, that the influence of temperature on bacterial inactivation by high-pressure homogenization is only through its effect on fluid viscosity. At initial temperatures > or =45 degrees C, corresponding to an outlet sample temperature >65 degrees C, the level of inactivation was higher than what would be predicted on the basis of the reduced viscosity at these temperatures, suggesting that under these conditions heat starts to contribute to cellular inactivation in addition to the mechanical effects that are predominant at lower temperatures. Second-order polynomial models were proposed to describe the impact of a high-pressure homogenization treatment of E. coli MG1655 as a function of pressure and temperature or as a function of pressure and viscosity. The pressure-viscosity inactivation model provided a better quality of fit of the experimental data and furthermore is more comprehensive and versatile than the pressure-temperature model because in addition to viscosity it implicitly incorporates temperature as a variable.     

THE VAPOR PRESSURE OF DOG'S BLOOD AT BODY TEMPERATURE

The vapor pressures of dog''s blood and blood plasma were determined at 37.5° by the dynamic method and the osmotic pressures calculated from the experimental data. The vapor pressures calculated from experimentally determined freezing point data agreed, within the experimental error, with the values obtained from direct measurement. The vapor pressure lowering produced by the colloid constituents of the blood was also determined and found to be minimal compared to that of the other constituents.     

Survival conditions of microorganisms under extremely severe environment]

The survival conditions of microorganisms under extremely severe environment are of interest in various areas of biology, sterilization, and space engineering, especially where resistance to microorganisms is concerned. Despite the interest, the resistance to microorganisms under extremely severe environment such as space environment or other planetary environment is not known well. In order to investigate survival conditions of microorganisms under extremely severe environment, surviving fractions for spores and vegetative cells of Bacillus subtilis were surveyed in various chemical species of atmosphere at various pressures and various temperatures, and the dependence on time for surviving fractions was examined. The results show: (i) Surviving fractions depend on chemical species of atmosphere. (ii) At high pressure and high temperature, surviving fractions are low and the resistance of spores is stronger than that of vegetative cells. (iii) Surviving fractions decrease as first-order reaction along with time elapsed.     

Effect of water vapor pressure on the thermal decomposition of 2-chloroethylphosphonic Acid

Decomposition of 2-chloroethylphosphonic acid (Ethephon) was studied in dried films at various water vapor pressures (0.6 to 86.9 millimeters Hg, 3.2 to 93.9% relative humidity) and temperatures (20, 30, 40, and 50 C) at pH 6.3 and 7.0. The rate of decomposition could be determined equally well by [¹⁴C]Ethephon or ethylene measurements. The rate increases at increasing water vapor pressures at a constant temperature and pH, up to an optimum. The optimum vapor pressure for decomposition approximately doubles for each 10 C increase. The activation energy for the decomposition reaction in water vapor pressures of 3.2 to 12 millimeters Hg is 8.7 and 14.3 kilocalories per mole at pH 6.3 and 7.0, respectively.     

Evaluation of manometric temperature measurement, a process analytical technology tool for freeze-drying: Part II measurement of dry-layer resistance

The purpose of this work was to study the factors that may cause systematic errors in the manometric temperature measurement (MTM) procedure used to determine product dry-layer resistance to vapor flow. Product temperature and dry-layer resistance were obtained using MTM software installed on a laboratory freeze-dryer. The MTM resistance values were compared with the resistance values obtained using the “vial method.” The product dry-layer resistances obtained by MTM, assuming fixed temperature difference (ΔT; 2°C), were lower than the actual values, especially when the product temperatures and sublimation rates were low, but with ΔT determined from the pressure rise data, more accurate results were obtained. MTM resistance values were generally lower than the values obtained with the vial method, particularly whenever freeze-drying was conducted under conditions that produced large variations in product temperature (ie, low shelf temperature, low chamber pressure, and without thermal shields). In an experiment designed to magnify temperature heterogeneity, MTM resistance values were much lower than the simple average of the product resistances. However, in experiments where product temperatures were homogenous, good agreement between MTM and “vial-method” resistances was obtained. The reason for the low MTM resistance problem is the fast vapor pressure rise from a few “warm” edge vials or vials with low resistance. With proper use of thermal shields, and the evaluation of ΔT from the data, MTM resistance data are accurate. Thus, the MTM method for determining dry-layer resistance is a useful tool for freeze-drying process analytical technology.     

Optimization of batch fermentor sterilization

This article presents a calculation procedure useful for the optimization and scale up of batch sterilization cycles in large-scale fermentors. This technique determines the sterilization temperature and hold-time necessary to minimize nutrient damage in a specific fermentor. The method can also be used for "scaledown" experiments to eliminate sterilization conditions as a scale up parameter. A method for the systematic evaluation of different sterilization conditions on product yield is also presented. This procedure is useful in determining if scale up of sterilization conditions is important for a given process. The validity of the techniques presented are supported by data showing significant yield improvements in a 1.2 x 10(5) L antibiotic fermentation.     

Study of semiconducting nanomaterials under pressure

The pressure induced structural and mechanical properties of nanocrystalline ZnO, ZnS, ZnSe, GaN, CoO, CdSe, CeO(2), SnO(2), SiC, c-BC(2)N, and β-Ga(2)O(3) with different grain sizes have been analyzed under high pressures. The molecular dynamics simulation model has been used to compute isothermal equation of state, volume collapse and bulk modulus of these materials in nano and bulk phases at ambient and high pressures and compared with the experimental data. It is evident from these calculations that the change in particle size affects directly the phase transition pressure and bulk modulus. The values of phase transition pressure and bulk modulus increase with decrease in grain size of the material. The equilibrium cell volume and volume collapse in parent phase is directly proportional to the grain size of the materials. Present results are in good agreement with experimental data. The model is able to explain these thermodynamic properties at varying temperatures and pressures successfully.     

Impact of high pressure freezing on DH5alpha Escherichia coli and red blood cells

The impact of high pressure and freezing on survivability of Escherichia coli and human red blood cells was evaluated to determine the utility of high-pressure transitions for preserving living cells. Based on microscopy and survivability, high pressures did not directly impact physical damage to living cells. E. coli studies showed that increased cell death is due to indirect phenomena with decreasing survivability at increasingly high pressures and exposure times. Pressurization rates up to 1.4kbar/min had negligible effects relative to exposures of >5min at high pressures.Both glycine and control of pH near 7.0 were successful in reducing the adverse impacts of high pressure. Survivability increased from <1% at 5min exposure to 2.1kbar of pressure to typical values >20%. The combination of glycine and the buffer salt led to even further improvements in survivability. Pressure changes were used to traverse temperature and pressures consistent with Ice I and Ice III phase boundaries of pure water.     

Theoretical and experimental studies on freezing point depression and vapor pressure deficit as methods to measure osmotic pressure of aqueous polyethylene glycol and bovine serum albumin solutions

For survival in adverse environments where there is drought, high salt concentration or low temperature, some plants seem to be able to synthesize biochemical compounds, including proteins, in response to changes in water activity or osmotic pressure. Measurement of the water activity or osmotic pressure of simple aqueous solutions has been based on freezing point depression or vapor pressure deficit. Measurement of the osmotic pressure of plants under water stress has been mainly based on vapor pressure deficit. However, differences have been noted for osmotic pressure values of aqueous polyethylene glycol (PEG) solutions measured by freezing point depression and vapor pressure deficit. For this paper, the physicochemical basis of freezing point depression and vapor pressure deficit were first examined theoretically and then, the osmotic pressure of aqueous ethylene glycol and of PEG solutions were measured by both freezing point depression and vapor pressure deficit in comparison with other aqueous solutions such as NaCl, KCl, CaCl(2), glucose, sucrose, raffinose, and bovine serum albumin (BSA) solutions. The results showed that: (1) freezing point depression and vapor pressure deficit share theoretically the same physicochemical basis; (2) theoretically, they are proportional to the molal concentration of the aqueous solutions to be measured; (3) in practice, the osmotic pressure levels of aqueous NaCl, KCl, CaCl(2), glucose, sucrose, and raffinose solutions increase in proportion to their molal concentrations and there is little inconsistency between those measured by freezing point depression and vapor pressure deficit; (4) the osmotic pressure levels of aqueous ethylene glycol and PEG solutions measured by freezing point depression differed from the values measured by vapor pressure deficit; (5) the osmotic pressure of aqueous BSA solution measured by freezing point depression differed slightly from that measured by vapor pressure deficit.     

Collapse temperature of freeze-dried Lactobacillus bulgaricus suspensions and protective media

Optimization of the freeze-drying process needs to characterize the physical state of frozen and dried products. A protocol to measure the collapse temperature of complex biological media such as concentrated lactic acid bacteria using freeze-drying microscopy was first elaborated. Afterward, aqueous solutions of one or several components as well as concentrated lactic acid bacterial suspensions were analyzed in order to study how the structure of these materials is degraded during freeze-drying. A similar behavior toward collapse was observed for all aqueous solutions, which was characterized by two temperatures: the "microcollapse" temperature (T(microc), beginning of a local loss of structure) and the "collapse" temperature (T(c), beginning of an overall loss of structure). For aqueous solutions, these two temperatures were close, differing by less than 3 degrees C. Nevertheless, when lactic acid bacteria were added to aqueous solutions, the collapse temperatures increased. Moreover, the interval between microcollapse and collapse temperatures became larger. Lactic acid bacterial cells gave a kind of "robustness" to the freeze-dried product. Finally, comparing glass transition, measured by differential scanning calorimetry (DSC) and collapse temperature for aqueous solutions with noncrystallizable solutes, showed that these values belonged to the same temperature range (differing by less than 5 degrees C). As suggested in the literature, the glass transition temperature can thus be used as a first approximation of the collapse temperature of these media. However, for lactic acid bacterial suspensions, because the difference between collapse and glass transition temperatures was about 10 degrees C, this approximation was not justified. An elegant physical appearance of the dried cakes and an acceptable acidification activity recovery were obtained, when applying operating conditions during freeze-drying in vials that allowed the product temperature to be maintained during primary drying at a level lower than the collapse temperature of lactic acid bacterial suspensions. Consequently, the collapse temperature T(c) was proposed as the maximal product temperature preserving the structure from macroscopic collapse and an acceptable biological activity of cells.     

Equilibrium temperature in a clump of bacteria heated in fluid

A theoretical model was developed and used to estimate quantitatively the "worst case", i.e., the longest, time to reach equilibrium temperature in the center of a clump of bacteria heated in fluid. For clumps with 10 to 10(6) cells heated in vapor, such as dry and moist air, and liquid fluids such as purees and juices, predictions show that temperature equilibrium will occur with sterilization temperatures up to 130 degrees C in under 0.02 s. Model development highlighted that the controlling influence on time for heating up the clump is the surface convection thermal resistance and that the internal conduction resistance of the clump mass is negligible by comparison. The time for a clump to reach equilibrium sterilization temperature was therefore decreased with relative turbulence (velocity) of the heating fluid, such as occurs in many process operations. These results confirm widely held suppositions that the heat-up time of bacteria in vapor or liquid is not significant with usual sterilization times.     

Equilibrium temperature in a clump of bacteria heated in fluid.

A theoretical model was developed and used to estimate quantitatively the "worst case", i.e., the longest, time to reach equilibrium temperature in the center of a clump of bacteria heated in fluid. For clumps with 10 to 10(6) cells heated in vapor, such as dry and moist air, and liquid fluids such as purees and juices, predictions show that temperature equilibrium will occur with sterilization temperatures up to 130 degrees C in under 0.02 s. Model development highlighted that the controlling influence on time for heating up the clump is the surface convection thermal resistance and that the internal conduction resistance of the clump mass is negligible by comparison. The time for a clump to reach equilibrium sterilization temperature was therefore decreased with relative turbulence (velocity) of the heating fluid, such as occurs in many process operations. These results confirm widely held suppositions that the heat-up time of bacteria in vapor or liquid is not significant with usual sterilization times.     

Temperature dependence of F-, D- and z-values used in steam sterilization processes

Aims:  To develop a model, based on microbiological principles, to safely optimize steam sterilization processes.
Methods and Results:  The minimum exposure time F for a decontamination process at a certain temperature is usually calculated from an empirical model with the decimal reduction time D and the temperature resistance coefficient z as parameters. These are implicitly assumed to be independent of temperature. Using a microbiological approach, it is shown that also D and z depend on temperature, indicating that the usual models provide only reliable results in a limited temperature region. The temperature dependence of F resulting from this approach describes the available experimental data very well. Safety margins to assure sterility can be included in a straightforward way.
Conclusions:  The results from the present approach can be used to safely optimize decontamination processes. The corresponding mathematical model can be implemented rather directly in process control systems.
Significance and Impact of the Study:  Our results show that for steam sterilization and disinfection processes the values of F predicted by the usual models largely underestimate the required minimum exposure times at temperatures below 120°C. This has important consequences for the optimization of such processes.     

Barophilic growth of bacteria from intestinal tracts of deep-sea invertebrates

Digestive tracts of abyssal scavenging amphipods and a deep-sea holothurian were examined for the presence of intestinal microflora capable of rapid proliferation under in situ pressures of 430 to 520 atmospheres (atm) and temperatures of 3–5°C. For two amphipod specimens, population doubling times of 5 and 6 hours were observed under in situ conditions, compared to 8 and 6 hours, respectively, at 1 atm. Growth enhancement under pressure was related inversely to initial population size and directly to concentration of available nutrient. In the case of the deposit-feeding holothurian, attached bacteria scraped from the intestinal lining showed a doubling time, under pressure, of 11 hours, compared to 36 hours for transient sediment bacteria that comprised the gut contents. These data suggest that deep-sea animals possess a commensal gut flora capable of responding to increased nutrient levels, via feeding of the host, without inhibition by the elevated hydrostatic pressures encountered in the deep ocean environment.     

Biomechanical analysis of fluid percussion model of brain injury

Fluid percussion injury (FPI) is a widely used experimental model for studying traumatic brain injury (TBI). However, little is known about how the brain mechanically responds to fluid impacts and how the mechanical pressures/strains of the brain correlate to subsequent brain damage for rodents during FPI. Hence, we developed a numerical approach to simulate FPI experiments on rats and characterize rat brain pressure/strain responses at a high resolution. A previous rat brain model was improved with a new hexahedral elements-based skull model and a new cerebrospinal fluid (CSF) layer. We validated the numerical model against experimentally measured pressures from FPI. Our results indicated that brain tissues under FPI experienced high pressures, which were slightly lower (10–20%) than input saline pressure. Interestingly, FPI was a mixed focus- and diffuse-type injury model with highest strains (12%) being concentrated in the ipsilateral cortex under the fluid-impact site and diffuse strains (5–10%) being spread to the entire brain, which was different from controlled cortical impact in which high strains decreased gradually away from the impact site.     

Effect of temperature and pressure on the protonation of glycine

Flow calorimetry has been used to study the interaction of glycine with protons in water at temperatures of 298.15, 323.15, and 348.15 K and pressures up to 12.50 MPa. By combining the measured heat for glycine solutions titrated with NaOH with the heat of ionization for water, the enthalpy of protonation of glycine is obtained. The reaction is exothermic at all temperatures and pressures studied. The effect of pressure on the enthalpy of reaction is very small. The experimental heat data are analyzed to yield equilibrium constant (K), enthalpy change (ΔH), and entropy change (ΔS) values for the protonation reaction as a function of temperature. These values are compared with those reported previously at 298.15 K. The ΔH and ΔS values increase (become more positive), whereas log K values decrease, as temperature increases. The trends for ΔH and ΔS with temperature are opposite to those reported previously for the protonation of several alkanolamines. However, log K values for proton interaction with both glycine and the alkanolamines decrease with increasing temperature. The effect of the nitrogen atom substituent on log K for protonation of glycine and alkanolamines is discussed in terms of changes in long-range and short-range solvent effects. These effects are used to explain the difference in ΔH and ΔS trends between glycine protonation and those found earlier for alkanolamine protonation.     

Temperature and phosphorus interacts in controlling the picoplankton carbon flux in the Adriatic Sea: an experimental versus field study

Temperature and phosphorus positively interacted in controlling picoplankton biomass production and its transfer towards higher trophic levels. Two complementary approaches (experimental and field study) indicated several coherent patterns: (1) the impact of temperature on heterotrophic bacteria was high at temperatures lower than 16°C and levelled off at higher temperatures, whereas this impact on autotrophic picoplankton was linear along the entire range of the investigated temperatures; (2) the addition of phosphorus increased the values of picoplankton production and grazing, but did not change the nature of their relationships with temperature substantially; (3) the picoplankton carbon flux towards higher trophic levels was larger during the warmer months (grazing by HNF dominated during the warmer period and by ciliates during the colder period) and also strengthened in conditions without phosphorus limitation; (4) the hypothesis that the available phosphorus can be better utilized at higher temperatures was confirmed for both autotrophic and heterotrophic picoplankton; (5) the hypothesis that the rise in temperature stimulates growth only in conditions of sufficient phosphorus was confirmed only for heterotrophic bacteria. Therefore, in the global warming scenario, an increase of the picoplankton carbon flux towards higher trophic levels can be expected in the Adriatic Sea, particularly under unlimited phosphorus conditions.     

Atmospheric vapor pressure deficit is critical in predicting growth response of “cool-season” grass <Emphasis Type="Italic">Festuca arundinacea</Emphasis> to temperature change

There is a lack of information on plant response to multifactor environmental variability including the interactive response to temperature and atmospheric humidity. These two factors are almost always confounded because saturated vapor pressure increases exponentially with temperature, and vapor pressure deficit (VPD) could have a large impact on plant growth. In this study using climate controlled mini-greenhouses, we examined the interacting influence of temperature and VPD on long-term growth of tall fescue (Festuca arundinacea Schreb), a cool season grass. From past studies it was expected that growth of tall fescue would decline with warmer temperatures over the range of 18.5–27°C, but growth actually increased markedly with increasing temperature when VPD was held constant. In contrast, growth declined in experiments where tall fescue was exposed to increasing VPD and temperature was held constant at 21°C. The inhibited growth appears to be in response to a maximum transpiration rate that can be supported by the tall fescue plants. The sensitivity to VPD indicates that if VPD remains stable in future climates as it has in the past, growth of tall fescue could well be stimulated rather than decreased by global warming in temperate climate zones.