Headspace Pressure Research Articles

Parallelization of photocatalytic gas-producing reactions.

High-throughput screening has been widely utilized in the pharmaceutical and manufacturing industry targeting the development of new molecules and materials for numerous applications. To enable more rapid progress in photocatalytic water-splitting reactions, the construction of high-throughput combinatorial photoreactors enabling the parallel optimization of relevant compositions under varieties of experimental conditions seems appropriate. This contribution describes a 16-photoreactor apparatus permitting the kinetic evaluation of photocatalytic gas-producing reactions using head-space pressure, gas chromatography, and mass spectrometry operating in parallel, illustrated with molecular-based homogeneous photocatalytic H2-generating compositions.

Effect of sugar on bread dough aeration during mixing

The aim of this study was to understand better dough aeration and in particular to determine how aeration during mixing differs between sugar containing and non-sugar containing dough through modelling and X-ray tomography. Batches of non-yeasted bread dough were mixed using both ‘plain’ and ‘strong’ flours with systematically varying granulated caster sugar formulations. A modified Tweedy 1-type mixer was used and mixer headspace pressure was controlled at various constant values to determine steady state dough aeration conditions. Pressure step-change experiments were used to determine aeration dynamics. A population balance model was fitted to the data to enable quantitative determination of the gas entrainment rate and disentrainment coefficient. X-ray computed tomography studies and uniaxial extensibility tests were also conducted to explore further the results of the aeration studies. This method enabled the model to be fitted to the aeration step-change response using only one parameter. The main effects of granulated sugar in dough were to increase the disentrainment coefficient and, to a lesser extent, to increase the entrainment rate. The combined effect of these was a reduction in dough gas content as granulated sugar content increased.

Development of an industrializable fermentation process for propionic acid production

Propionic acid (PA) is a short-chain fatty acid with wide industrial application including uses in pharmaceuticals, herbicides, cosmetics, and food preservatives. As a three-carbon building block, PA also has potential as a precursor for high-volume commodity chemicals such as propylene. Currently, most PA is manufactured through petrochemical routes, which can be tied to increasing prices and volatility due to difficulty in demand forecasting and feedstock availability. Herein described are research advancements to develop an industrially feasible, renewable route to PA. Seventeen Propionibacterium strains were screened using glucose and sucrose as the carbon source to identify the best platform strain. Propionibacterium acidipropionici ATCC 4875 was selected as the platform strain and subsequent fermentation optimization studies were performed to maximize productivity and yield. Fermentation productivity was improved three-fold to exceed 2 g/l/h by densifying the inoculum source. Byproduct levels, particularly lactic and succinic acid, were reduced by optimizing fermentor headspace pressure and shear. Following achievement of commercially viable productivities, the lab-grade medium components were replaced with industrial counterparts to further reduce fermentation costs. A pure enzymatically treated corn mash (ECM) medium improved the apparent PA yield to 0.6 g/g (PA produced/glucose consumed), but it came at the cost of reduced productivity. Supplementation of ECM with cyanocobalamin restored productivity to near lab-grade media levels. The optimized ECM recipe achieved a productivity of 0.5 g/l/h with an apparent PA yield of 0.60 g/g corresponding to a media cost <1 USD/kg of PA. These improvements significantly narrow the gap between the fermentation and incumbent petrochemical processes, which is estimated to have a manufacturing cost of 0.82 USD/kg in 2017.

Focus on sensors and detectors

Focus on sensors and detectors

Sensor for headspace pressure and H2O concentration measurements in closed vials by tunable diode laser absorption spectroscopy

The concentration of H2O and the pressure in the headspace of vials are simultaneously measured by a tunable diode laser sensor based on absorption spectroscopy techniques. The 7168.437cm−1 spectral line of H2O is chosen as the sensing transition for its strong absorption strength and being reasonably far away from its neighboring molecular transitions. In order to prevent interference absorption by ambient water vapor in the room air, a difference between the measured signal and the referenced signal is used to calculate the pressure and H2O concentration in the headspace of vials, eliminating the need for inert gas purges and calibration with known gas. The validation of the sensor is conducted in a static vial, yielding an accuracy of 1.23% for pressure and 3.81% for H2O concentration. The sensitivity of the sensor is estimated to be about 2.5Torr for pressure and 400ppm for H2O concentration over a 3cm absorption path length respectively. Accurate measurements for commercial freeze-dried products demonstrate the in-line applications of the sensor for the pharmaceutical industry.

Bread dough aeration dynamics during pressure step-change mixing: Studies by X-ray tomography, dough density and population balance modelling

Industrial bread dough mixing often involves a period of mixing under high headspace pressure to enhance oxygen availability, followed by a period of partial vacuum to favourably manipulate the final bubble size distribution. This paper presents the results of a study using X-ray tomography to measure the gas bubble size distribution in dough samples over the course of a pressure step-change mix. The first objective of the current work was to measure bubble size distributions at points throughout a pressure-step dough mixing process using a non-synchrotron X-ray source. The second objective was to fit a simplified population balance model to the measured size distributions. The third objective was to use the data set and fitted model to explore the validity of the assumptions within the simplified model and to consolidate understanding of underlying aeration and mixing phenomena and the resultant process dynamics. It was found that the dynamic changes in the bubble size distribution of a bread dough during pressure-step mixing could be accurately measured using a laboratory X-ray source. The response of the cumulative dough voidage to a pressure-step change during mixing could be reproduced very well using the simplified population balance model (which assumes: all entrained bubbles are the same size, no bubble break-up or coalescence, and likelihood of bubble disentrainment is proportional to bubble volume). The measured response of the bubble number density and mean volume agreed reasonably well with that predicted by the simplified model (with parameters fitted to only cumulative voidage data). It is demonstrated that the decrease in number density following a pressure step-decrease is much more short-lived than the decrease in size which is permanent.

Pressure‐monitored headspace analysis combined with compound‐specific isotope analysis to measure isotope fractionation in gas‐producing reactions

Processes that lead to pressure changes in closed experimental systems can dramatically increase the total uncertainty in enrichment factors (ε) based on headspace analysis and compound-specific isotope analysis (CSIA). We report: (1) A new technique to determine ε values for non-isobaric processes, and (2) a general approach to evaluate the experimental error in calculated ε values. ε values were determined by monitoring the change in headspace pressure from the production of CO2 in a decarboxylation reaction using a pressure gauge and measuring the δ(13) C values using CSIA. The statistical error was assessed over shorter reaction progress intervals to evaluate the impact of experimental error on the total uncertainty associated with calculated ε values. As an alternative to conventional compositional analysis, calculation of CO2 produced during the reaction monitored with a pressure gauge resulted in rate constants and ε values with improved correlation coefficients and confidence intervals for a non-isobaric process in a closed system. Further, statistical evaluation of the ε values as a function of reaction progress showed that uncertainty in data points for reaction progress (f) at late stages of the reaction can have a significant impact on the reported ε value. Pressure-monitored headspace analysis reduces the uncertainty associated with monitoring the reaction progress (f) based on estimating substrate removal and headspace dilution during sampling. Statistical calculations over shorter intervals should be used to evaluate the total error for reported ε values.

Drying Process Optimization for an API Solvate Using Heat Transfer Model of an Agitated Filter Dryer

Drying an early stage active pharmaceutical ingredient candidate required excessively long cycle times in a pilot plant agitated filter dryer. The key to faster drying is to ensure sufficient heat transfer and minimize mass transfer limitations. Designing the right mixing protocol is of utmost importance to achieve efficient heat transfer. To this order, a composite model was developed for the removal of bound solvent that incorporates models for heat transfer and desolvation kinetics. The proposed heat transfer model differs from previously reported models in two respects: it accounts for the effects of a gas gap between the vessel wall and solids on the overall heat transfer coefficient, and headspace pressure on the mean free path length of the inert gas and thereby on the heat transfer between the vessel wall and the first layer of solids. A computational methodology was developed incorporating the effects of mixing and headspace pressure to simulate the drying profile using a modified model framework within the Dynochem software. A dryer operational protocol was designed based on the desolvation kinetics, thermal stability studies of wet and dry cake, and the understanding gained through model simulations, resulting in a multifold reduction in drying time.

Container/Closure Integrity Testing and the Identification of a Suitable Vial/Stopper Combination for Low-Temperature Storage at -80 C

It was recently found that after storage of a live viral vaccine at -80 °C in glass vials closed with rubber stoppers, a phenomenon was revealed which had not been observed before with other viral products stored at -20 °C: overpressure in the vials. As this phenomenon poses a serious safety problem for medical personnel as well as for the product itself, an investigation was initiated to identify the root cause of the overpressure. After exclusion of possible root causes (differences in air temperature or atmospheric air pressure during filling and quality control testing, outgassing from the formulation buffer) the remaining hypothesis involved a possible container closure integrity issue at low temperature. The glass transition temperatures (T(g)) of many rubber stopper formulations are in the range -55 to -70 °C. At storage temperatures below T(g), the rubber stopper loses its elastic properties and there is a risk that the seal integrity of the vial could be compromised. Loss of seal integrity of the vials near storage temperatures of -80 °C would result in an ingress of cold dense gas into the vial headspace. After removal of the vials from storage at -80 °C, the rubber stoppers could regain their elastic properties and the vials would quickly reseal, thereby trapping the ingressed gas, which leads to overpressure in the vial headspace. Nondestructive laser-based headspace analysis was used to investigate the maintenance of container closure integrity as a function of the filling and capping/crimping process, storage and transport conditions, and vial/stopper designs. This analytical method is based on frequency modulation spectroscopy (FMS) and can be used for noninvasive headspace measurements of headspace pressure and headspace gas composition. Changes in the vial headspace composition and/or pressure are a clear marker for vials that have lost container closure integrity. After storage of a live viral vaccine at -80 °C in glass vials closed with rubber stoppers, overpressure in some of the vials was observed, posing a serious safety problem for medical personnel as well as for the product. A working hypothesis to explain this phenomenon involved a possible container closure integrity issue at these low temperatures. The glass transition temperatures (T(g)) of many rubber stopper formulations are in the range -55 to -70 °C. At storage temperatures below T(g), the rubber stopper loses its elastic properties, resulting in compromised seal integrity of the vial and ingress of cold dense gas into the vial headspace. Upon thawing, the rubber stoppers regain their elastic properties and the vials quickly reseal, thereby trapping the ingressed gas, which leads to overpressure in the vial headspace. Nondestructive, laser-based headspace analysis, which is able to detect changes in headspace pressure and gas composition, was used to investigate the maintenance of container closure integrity. Changes in the vial headspace composition and/or pressure are a clear marker for vials that have lost container closure integrity.

The effect of low pressure and mixing on biological hydrogen production via anaerobic fermentation

Two-stage anaerobic digesters are ideally suited for hydrogen production because primary fermentation is separated from methanogenesis, allowing for optimization of both processes. At low hydrogen partial pressures, hydrogen forming reactions become thermodynamically favorable and there is a shift in fermentation toward the production of acetate and away from butyrate, alcohols, and solvents. Increased acetate yields in first stage reactors enhance acetoclastic methanogenesis in second stage reactors. We evaluated the effects of headspace pressure and reactor mixing, two parameters that can be controlled at the industrial scale, on hydrogen production and fermentation products using anaerobic batch reactors. Control of methanogenesis was achieved with low pH caused by high loading of non-fat powered milk (4 gVS L−1). In mixed reactors, hydrogen yields increased significantly from 87.5 to 121.4 ml gVS−1 (p = 0.056) and the acetate to butyrate ratio increased from 0.44 to 0.68 (p = 0.11). Control of headspace pressure below atmospheric levels did not have a significant effect on gas production. These results demonstrate the rapid production of hydrogen and inhibition of methanogenesis via substrate loading and mixing, and provide a practical method for enhancing energy recovery in two-stage anaerobic digesters.

Model for Estimating CO2Concentration in Package Headspace of Microbiologically Perishable Food

Levels of carbon dioxide gas, a metabolite of microbial growth, have been reported to parallel the onset of microbial spoilage and may be used as a convenient index for a packaged food’s shelf life. This study aimed to establish a kinetic model of CO₂ production from perishable food for the potential use for shelf life control in the food supply chain. Aerobic bacterial count and package CO₂ concentration were measured during the storage of seasoned pork meat at four temperatures (0, 5, 10 and 15℃), and their interrelationship was investigated to establish a mathematical model. The microbial growth at constant temperature was described by using model of Baranyi and Roberts. CO₂ production from the stored food could be explained by taking care of its yield and maintenance factors linked to the microbial growth. By establishing the temperature dependence of the microbial growth and CO₂ yield factor, CO₂ partial pressure or concentration in package headspace could be estimated to a limited extent, which is helpful for controlling the shelf life under constant and dynamic temperature conditions. Application and efficacy of the model needs to be improved with further refinement in the model.

Evaluation of the energy efficiency of enzyme fermentation by mechanistic modeling

Modeling biotechnological processes is key to obtaining increased productivity and efficiency. Particularly crucial to successful modeling of such systems is the coupling of the physical transport phenomena and the biological activity in one model. We have applied a model for the expression of cellulosic enzymes by the filamentous fungus Trichoderma reesei and found excellent agreement with experimental data. The most influential factor was demonstrated to be viscosity and its influence on mass transfer. Not surprisingly, the biological model is also shown to have high influence on the model prediction. At different rates of agitation and aeration as well as headspace pressure, we can predict the energy efficiency of oxygen transfer, a key process parameter for economical production of industrial enzymes. An inverse relationship between the productivity and energy efficiency of the process was found. This modeling approach can be used by manufacturers to evaluate the enzyme fermentation process for a range of different process conditions with regard to energy efficiency.

Investigating Liquid Leak from Pre-Filled Syringes upon Needle Shield Removal: Effect of Air Bubble Pressure

This study is to investigate the effect of headspace air pressure in pre-filled syringes on liquid leak (dripping) from the syringe needle upon needle shield removal. Drip tests to measure drip quantity were performed on syringes manually filled with 0.5 or 1.0 mL of various aqueous solutions. Parameters assessed included temperature (filling and test), bulk storage conditions (tank pressure and the type of the pressurized gas), solution composition (pure water, 0.9% sodium chloride, and a monoclonal antibody formulation), and testing procedures. A headspace pressure analyzer was used to verify the drip test method. Results suggested that leakage is indeed caused by headspace pressure increase, and the temperature effect (ideal gas expansion) is a major, but not the only, factor. The dissolved gases in the liquid bulk prior to or during filling may contribute to leakage, as these gases could be released into the headspace due to solubility changes (in response to test temperature and pressure conditions) and cause pressure increase. Needle shield removal procedures were found to cause dripping, but liquid composition played little role. Overall, paying attention to the processing history (pressure and temperature) of the liquid bulk is the key to minimize leakage. The headspace pressure could be reduced by decreasing liquid bulk storage pressure, filling at a higher temperature, or employing lower solubility gas (e.g., helium) for bulk transfer and storage. Leakage could also be mitigated by simply holding the syringe needle pointing upward during needle shield removal. Substantial advances in pre-filled syringe technology development, particularly in syringe filling accuracy, have been made. However, there are factors, as subtle as how the needle shield (or tip cap) is removed, that may affect dosing accuracy. We recently found that upon removal of the tip cap from a syringe held vertically with needle pointed downwards, a small amount of solution, up to 3-4% of the 1 mL filled volume or higher for filled volume of <1 mL, leaked out from the needle. This paper identified the root causes of this problem and offered solutions from the perspectives of the syringe fill process and the end user procedure. The readers will benefit from this paper by understanding how each process step prior to and during syringe filling may affect delivery performance of the pre-filled syringe device.

Measuring and Predicting Head Space Pressure during Retorting of Thermally Processed Foods

Traditional metal cans and glass jars have been the mainstay in thermally processed canned foods for more than a century, but are now sharing shelf space with increasingly popular flexible pouches and semi-rigid trays. These flexible packages lack the strength of metal cans and glass jars, and need greater control of external retort pressure during processing. Increasing internal package pressure without counter pressure causes volumetric expansion, putting excessive strain on package seals that may lead to serious container deformation and compromised seal integrity. The primary objective of this study was to measure internal pressure build-up within a rigid air-tight container (module) filled with various model food systems undergoing a retort process in which internal product temperature and pressure, along with external retort temperature and pressure, were measured and recorded at the same time. The pressure build-up in the module was compared with the external retort pressure to determine the pressure differential that would cause package distortion in the case of a flexible package system. The secondary objective was to develop mathematical models to predict these pressure profiles in response to known internal temperature and initial and boundary conditions for the case of the very simplest of model food systems (pure water and aqueous saline and sucrose solutions), followed by food systems of increasing compositional complexity (green beans in water and sweet peas in water). Results showed that error between measured and predicted pressures ranged from 2% to 4% for water, saline, and green beans, and 7% to 13% for sucrose solution and sweet peas. Flexible packages have limited strength, and need more accurate and closer control of retort pressure during processing. The package becomes more flexible as it heats and might expand with increasing internal pressure that may cause serious deformation or rupture if not properly controlled and/or counterbalanced with external retort pressure. This article describes methods for determining exactly what the retort pressure profile will need to be to avoid this problem during retorting, and mathematical models to predict these pressures in response to known internal temperature and initial/boundary conditions.

Linear Correlation between Online Capacitance and Offline Biomass Measurement up to High Cell Densities in Escherichia coli Fermentations in a Pilot-Scale Pressurized Bioreactor

To yield high concentrations of protein expressed by genetically modified Escherichia coli, it is important that the bacterial strains are cultivated to high cell density in industrial bioprocesses. Since the expressed target protein is mostly accumulated inside the E. coli cells, the cellular product formation can be directly correlated to the bacterial biomass concentration. The typical way to determine this concentration is to sample offline. Such manual sampling, however, wastes time and is not efficient for acquiring direct feedback to control a fedbatch fermentation. An E. coli K12-derived strain was cultivated to high cell density in a pressurized stirred bioreactor on a pilot scale, by detecting biomass concentration online using a capacitance probe. This E. coli strain was grown in pure minimal medium using two carbon sources (glucose and glycerol). By applying exponential feeding profiles corresponding to a constant specific growth rate, the E. coli culture grew under carbon-limited conditions to minimize overflow metabolites. A high linearity was found between capacitance and biomass concentration, whereby up to 85 g/L dry cell weight was measured. To validate the viability of the culture, the oxygen transfer rate (OTR) was determined online, yielding maximum values of 0.69 mol/l/h and 0.98 mol/l/h by using glucose and glycerol as carbon sources, respectively. Consequently, online monitoring of biomass using a capacitance probe provides direct and fast information about the viable E. coli biomass generated under aerobic fermentation conditions at elevated headspace pressures.

Biohydrogen production in alkalithermophilic conditions: Thermobrachium celere as a case study

In the present work the hydrogenesis in the anaerobic alkalithermophilic bacterium Thermobrachium celere was studied. The impact of several factors on hydrogen production during glucose fermentation was investigated in batch conditions. The optimal hydrogen production occurred at pH 67 °C 8.2 with phosphate buffer concentration of 50 mM. Hydrogen yield reached the highest value of 3.36 mol H 2/mol glucose when the partial pressure in the gas headspace was reduced. Supplementation of nitrogen sources and iron affected hydrogen production. Under optimized conditions, the maximum H 2 accumulation and H 2 production rate were estimated to be respectively 124.3 mmol H 2/l culture and 20.7 mmol H 2/l/h. Considering the efficient and rapid hydrogen evolution, and the ability to grow in extreme environments, T. celere might be a good candidate for biohydrogen production in open (non-sterile) bioprocess system.

A novel ‘fizziness’ indicator

A novel colourimetric 'fizziness' indicator is described which changes colour depending on the headspace pressure of carbon dioxide above a carbonated liquid.

La culture et l’antibiogramme de mycobactéries sur automate VersaTREK

Objectives Our laboratory of microbiology use the VersaTREK/ESP Culture System II for the isolation of mycobacteria. In this study, we evaluate this system for the analysis carried out between April 2009 and March 2010. Method The Myco bottles are supplemented with growth supplements and an antimicrobial agent solution prior to inoculation with the specimen. The technology of the VersaTREK/ESP Culture System II is based on the detection of headspace pressure changes within a sealed bottle. It monitors changes in either gas production or gas consumption due to microbial growth. A special algorithm has been developed for detection of very slow growing mycobacteria. The bottles are incubated during 42 days. Meanwhile a solid medium is inoculated too. All specimen types can be analysed with this system. Results Compared to solid culture, the time needed for detection of positive cultures was significantly shorter for the VersaTREK with a good recovery rate. For isolates recovered in both systems, mean time of detection is respectively 19.1 and 35.6 days for liquid and solid cultures. Mycobacteria identification may be determined using nucleic acid probs directly in Myco VersaTREK or in the solid medium. The susceptibility test of Mycobacteria tuberculosis complex is obtained between six and 13 days for rifampin, isoniazid, ethambutol, streptomycin and pyrasinamide. Conclusion This system offers a faster diagnosis and is an alternative to other instruments using liquid culture.

High cell-density processes in batch mode of a genetically engineered Escherichia coli strain with minimized overflow metabolism using a pressurized bioreactor

A common method to minimize overflow metabolism and to enable high cell-density is to operate microbial processes in fed-batch mode under carbon-limiting conditions. This requires sophisticated process control schemes with expensive hardware equipment and software and well-characterized processes parameters. To generate high-cell density, a more simplified strategy would be beneficial. Therefore, a genetically engineered Escherichia coli strain with a modified glucose uptake system was cultivated in batch mode. In the applied strain, the usual phosphotransferase system of a K12-derived strain was inactivated, while the galactose permease system was amplified. Upon cultivating this E. coli strain in pure minimal media, the acetate concentration did not exceed values of 0.35 g L −1, even when the batch fermentation was started with a glucose concentration of 130 g L −1. Finally, maximum biomass concentrations of 48 g L −1 dry cell weight and maximum space-time yields of 2.10 g L −1 h −1 were reached. To provide an unlimited growth under fully aerobic conditions (DOT > 30%) at comparatively low values for specific power input (3–4 kW m −3), a pressurized bioreactor was used. Consequentially, to our knowledge, this study using a bioreactor with elevated headspace pressure generate the highest oxygen transfer rate (451 mmol L −1 h −1) ever reached in batch cultivations.

Microbially Enhanced Carbon Capture and Storage by Mineral-Trapping and Solubility-Trapping

The potential of microorganisms for enhancing carbon capture and storage (CCS) via mineral-trapping (where dissolved CO(2) is precipitated in carbonate minerals) and solubility trapping (as dissolved carbonate species in solution) was investigated. The bacterial hydrolysis of urea (ureolysis) was investigated in microcosms including synthetic brine (SB) mimicking a prospective deep subsurface CCS site with variable headspace pressures [p(CO(2))] of (13)C-CO(2). Dissolved Ca(2+) in the SB was completely precipitated as calcite during microbially induced hydrolysis of 5-20 g L(-1) urea. The incorporation of carbonate ions from (13)C-CO(2) ((13)C-CO(3)(2-)) into calcite increased with increasing p((13)CO(2)) and increasing urea concentrations: from 8.3% of total carbon in CaCO(3) at 1 g L(-1) to 31% at 5 g L(-1), and 37% at 20 g L(-1). This demonstrated that ureolysis was effective at precipitating initially gaseous [CO(2)(g)] originating from the headspace over the brine. Modeling the change in brine chemistry and carbonate precipitation after equilibration with the initial p(CO(2)) demonstrated that no net precipitation of CO(2)(g) via mineral-trapping occurred, since urea hydrolysis results in the production of dissolved inorganic carbon. However, the pH increase induced by bacterial ureolysis generated a net flux of CO(2)(g) into the brine. This reduced the headspace concentration of CO(2) by up to 32 mM per 100 mM urea hydrolyzed because the capacity of the brine for carbonate ions was increased, thus enhancing the solubility-trapping capacity of the brine. Together with the previously demonstrated permeability reduction of rock cores at high pressure by microbial biofilms and resilience of biofilms to supercritical CO(2), this suggests that engineered biomineralizing biofilms may enhance CCS via solubility-trapping, mineral formation, and CO(2)(g) leakage reduction.