The stabilisation of purified, reconstituted P-glycoprotein by freeze drying with disaccharides

The drug efflux pump P-glycoprotein (P-gp) (ABCB1) confers multidrug resistance, a major cause of failure in the chemotherapy of tumours, exacerbated by a shortage of potent and selective inhibitors. A high throughput assay using purified P-gp to screen and characterise potential inhibitors would greatly accelerate their development. However, long-term stability of purified reconstituted ABCB1 can only be reliably achieved with storage at -80 °C. For example, at 20 °C, the activity of ABCB1 was abrogated with a half-life of <1 day. The aim of this investigation was to stabilise purified, reconstituted ABCB1 to enable storage at higher temperatures and thereby enable design of a high throughput assay system. The ABCB1 purification procedure was optimised to allow successful freeze drying by substitution of glycerol with the disaccharides trehalose or maltose. Addition of disaccharides resulted in ATPase activity being retained immediately following lyophilisation with no significant difference between the two disaccharides. However, during storage trehalose preserved ATPase activity for several months regardless of the temperature (e.g. 60% retention at 150 days), whereas ATPase activity in maltose purified P-gp was affected by both storage time and temperature. The data provide an effective mechanism for the production of resilient purified, reconstituted ABCB1.

Drying of biomass for second generation synfuel production

Drying is a major and challenging step in the pre-treatment of biomass for production of second generation synfuels for transport. The biomass feedstocks are mostly wet and need to be dried from 30 to 60 wt% moisture content to about 10-15 wt%. The present survey aims to define and evaluate a few of the most promising optimised concepts for biomass pre-treatment scheme in the production of second generation synfuels for transport. The most promising commercially available drying processes were reviewed, focusing on the applications, operational factors and emissions of dryers. The most common dryers applied now for biomass in bio-energy plants are direct rotary dryers, but the use of steam drying techniques is increasing. Steam drying systems enable the integration of the dryer to existing energy sources. In addition to integration, emissions and fire or explosion risks have to be considered when selecting a dryer for the plant. In steam drying there will be no gaseous emissions, but the aqueous effluents need often treatment. Concepts for biomass pre-treatment were defined for two different cases including a large-scale wood-based gasification synfuel production and a small-scale pyrolysis process based on wood chips and miscanthus bundles. For the first case a pneumatic conveying steam dryer was suggested. In the second case the flue gas will be used as drying medium in a direct or indirect rotary dryer.

Mass transfer in fluidized bed drying of moist particulate

Bubbling fluidized bed technology is one of the most effective mean for interaction between solid and gas flow, mainly due to its good mixing and high heat and mass transfer rate. It has been widely used at a commercial scale for drying of grains such as in pharmaceutical, fertilizers and food industries. When applied to drying of non-pours moist solid particles, the water is drawn-off driven by the difference in water concentration between the solid phase and the fluidizing gas. In most cases, the fluidizing gas or drying agent is air. Despite of the simplicity of its operation, the design of a bubbling fluidized bed dryer requires an understanding of the combined complexity in hydrodynamics and the mass transfer mechanism. On the other hand, reliable mass transfer coefficient equations are also required to satisfy the growing interest in mathematical modelling and simulation, for accurate prediction of the process kinetics. This chapter presents an overview of the various mechanisms contributing to particulate drying in a bubbling fluidized bed and the mass transfer coefficient corresponding to each mechanism. In addition, a case study on measuring the overall mass transfer coefficient is discussed. These measurements are then used for the validation of mass transfer coefficient correlations and for assessing the various assumptions used in developing these correlations.

Freezing process optimisation of pharmaceutical formulations for freeze drying

The body of work presented in this thesis are in three main parts: [1] the effect of ultrasound on freezing events of ionic systems, [2] the importance of formulation osmolality in freeze drying, and [3] a novel system for increasing primary freeze drying rate. Chapter 4 briefly presents the work on method optimisation, which is still very much in its infancy. Aspects of freezing such as nucleation and ice crystal growth are strongly related with ice crystal morphology; however, the ice nucleation process typically occurs in a random, non-deterministic and spontaneous manner. In view of this, ultrasound, an emerging application in pharmaceutical sciences, has been applied to aid in the acceleration of nucleation and shorten the freezing process. The research presented in this thesis aimed to study the effect of sonication on nucleation events in ionic solutions, and more importantly how sonication impacts on the freezing process. This work confirmed that nucleation does occur in a random manner. It also showed that ultrasonication aids acceleration of the ice nucleation process and increases the freezing rate of a solution. Cryopreservation of animal sperm is an important aspect of breeding in animal science especially for endangered species. In order for sperm cryopreservation to be successful, cryoprotectants as well as semen extenders are used. One of the factors allowing semen preservation media to be optimum is the osmolality of the semen extenders used. Although preservation of animal sperm has no relation with freeze drying of pharmaceuticals, it was used in this thesis to make a case for considering the osmolality of a formulation (prepared for freeze drying) as a factor for conferring protein protection against the stresses of freeze drying. The osmolalities of some common solutes (mostly sugars) used in freeze drying were determined (molal concentration from 0.1m to 1.2m). Preliminary investigation on the osmolality and osmotic coefficients of common solutes were carried out. It was observed that the osmotic coefficient trend for the sugars analysed could be grouped based on the types of sugar they are. The trends observed show the need for further studies to be carried out with osmolality and to determine how it may be of importance to protein or API protection during freeze drying processes. Primary drying is usually the longest part of the freeze drying process, and primary drying times lasting days or even weeks are not uncommon; however, longer primary drying times lead to longer freeze drying cycles, and consequently increased production costs. Much work has been done previously by others using different processes (such as annealing) in order to improve primary drying times; however, these do not come without drawbacks. A novel system involving the formation of a frozen vial system which results in the creation of a void between the formulation and the inside wall of a vial has been devised to increase the primary freeze drying rate of formulations without product damage. Although the work is not nearly complete, it has been shown that it is possible to improve and increase the primary drying rate of formulations without making any modifications to existing formulations, changing storage vials, or increasing the surface area of freeze dryer shelves.

Spray drying lactoferrin produces inhalable antimicrobial particles

The use of antimicrobial peptides and proteins as potential therapeutic agents in the management of multi-drug resistant infections is considered an attractive concept especially since such compounds should theoretically have low immunogenicity, high bioavailability with negligible toxicity. In this study we investigated the potential of developing a dry powder inhaler formulation of lactoferrin (a multifunctional iron binding protein). To achieve this, the protein was spray dried from a water only feedstock with suitably adjusted spray drying parameters. The particle size, degree of crystallinity, moisture content and yield of the spray dried powders along with the minimum bactericidal concentration (MBC) against Pseudomonas aeruginosa strain PAO1, were assessed. Dry powder inhaler formulations were prepared, and in vitro assessment studies using the multistage impinger were carried out to assess the aerosolisation performance of the formulations. Data obtained indicate that spray dried lactoferrin retains activity against biofilms and may be successfully employed in the treatment of chronic airway infections.

Physicochemical characterization of grewia polysaccharide gum:effect of drying method

Grewia polysaccharide gum, a potential pharmaceutical excipient was extracted from the inner stem bark of Grewia mollis, thereupon drying was achieved by three techniques: air-drying, freeze-drying and spray-drying. Analysis of the monosaccharide composition including 1H and 13C NMR spectroscopic analysis of the polysaccharide gum was carried out. The effect of the drying methods on the physicochemical properties of the gum was evaluated by Fourier transformed infra-red (FT-IR) spectroscopy, solid-state 13C nuclear magnetic resonance (NMR) spectroscopy, X-ray photoelectron spectroscopy (XPS), thermogravimetric analysis, differential scanning calorimetry and gel permeation chromatography. Monosaccharide sugar analysis revealed that the gum is composed of glucose, rhamnose, galactose, arabinose and xylose as the main neutral sugars. These were supported by the results from 1H and 13C NMR spectroscopic analysis. FT-IR and solid-state NMR results indicated that drying technique has little effect on the structure of the polysaccharide gum but XPS showed that surface chemistry of the gum varied with drying methods. Thermogravimetric analyses showed that oxidation onset varied according to the drying method. The molecular weight was also dependent on the drying technique. For industrial extrapolation, air-drying may be preferable to spray-drying and freeze-drying when relative cost, product stability and powder flow are required, for example in tablet formulation. © 2010 Elsevier Ltd. All rights reserved.

Influence of parameters of drying on laser induced diffuse reflectance of banana discs

Banana discs of 1 cm thickness were immersed into different antioxidant solutions to slow down potentially disturbing discoloration during drying. Samples were randomly split into 8 groups according to the 2^p experimental design. Two antioxidant solutions with 1.66% and 4.59% ascorbic acid, two levels of drying temperature with 50°C and 80°C, two levels of drying time with 6h and 8h were used or adjusted. Laser diodes of seven wavelengths (532, 635, 650, 780, 808, 850 and 1064 nm) were selected to illuminate the surface and light penetration pattern was evaluated on the basis of radial profiles. Profiles acquired at three wavelengths (532, 635 and 650 nm) were found to respond sensitively to adjusted parameters. As a result of drying, intensity decay was observed to move closer to incident point. Significant effect (p<0.01) of temperature, drying time and their interaction was found on extracted descriptive attributes of intensity profiles: full width at half maximum (FWHM), distance of inflection point (DIP) and slope of logarithmic decay (SLD). Beyond their presence, antioxidant concentration was neutral factor without significant contribution to the model. Results were in agreement with reference spectroscopic measurements, especially with NDVI index. Promising results suggest that evaluated method might be suitable for monitoring purposes during drying of fruits.

Drying Times: Integrating Citizen Science to Examine Survival of Florida Largemouth Bass in a Coastal Refuge Habitat

In aquatic systems refuge habitats increase resistance to drying events and are necessary for maintaining populations in disturbed environments. However, reduced water availability and altered flow regimes threaten the existence and function of these habitats. To test refuge function I conducted a capture-mark-recapture (CMR) study, integrating citizen science angler sampling into fisheries-independent methods. The objectives of this study were twofold: 1) To determine the contribution of citizen science anglers to improving CMR research, and 2.) to quantify apparent survival of Florida Largemouth Bass, Micropterus salmoides floridanus, in a coastal refuge habitat across multiple years of drying severity. The inclusion of angler sampling was determined to be an effective and feasible method for increasing capture probability. Apparent survival of Florida Bass varied among hydrologic periods with lowest survival when marshes functionally dried (< 10 cm). Overall mortality from drying events increased with the duration of marsh drying upstream.

Stream drying drives microbial ammonia oxidation and first-flush nitrate export

Funded by Spanish National Research Council (CSIC). Grant Number: CGL2012-32747 MINECO. Grant Numbers: CGL2012-32747, CGL2011-30590-CO2-02 EU Commission. Grant Number: 244121 FP7

Drying kinetic analysis of municipal solid waste using modified page model and pattern search method

This work studied the drying kinetics of the organic fractions of municipal solid waste (MSW) samples with different initial moisture contents and presented a new method for determination of drying kinetic parameters. A series of drying experiments at different temperatures were performed by using a thermogravimetric technique. Based on the modified Page drying model and the general pattern search method, a new drying kinetic method was developed using multiple isothermal drying curves simultaneously. The new method fitted the experimental data more accurately than the traditional method. Drying kinetic behaviors under extrapolated conditions were also predicted and validated. The new method indicated that the drying activation energies for the samples with initial moisture contents of 31.1 and 17.2 % on wet basis were 25.97 and 24.73 kJ mol−1. These results are useful for drying process simulation and industrial dryer design. This new method can be also applied to determine the drying parameters of other materials with high reliability.

Evaporative Moisture Loss from Heterogeneous Stone: Material- Environment Interactions During Drying

The complexities of evaporation from structurally and mineralogically heterogeneous sandstone (Locharbriggs Sandstone) are investigated through a laboratory-based experiment in which a variety of environmental conditions are simulated. Data reported demonstrate the significance of material-environment interactions on the spatial and temporal variability of evaporative dynamics. Evaporation from porous stone is determined by the interplay between environmental, material and solution properties, which govern the rate and mode by which water is transmitted to, and subsequently removed from, an evaporating surface. Initially evaporation is marked by high rates of moisture loss controlled by external atmospheric conditions; then, when a critical level of surface moisture content is reached, hydraulic continuity between the stone surface and subsurface is disrupted and the drying front recedes
beneath the surface, evaporation rates decrease and are controlled by the ability of the material to transport water vapour to the surface. Pore size distribution and connectivity, as well as other material properties, control the timing of each stage of evaporation and the nature of the transition.

These experimental data highlight the complexity of evaporation, demonstrating that different regions of the same stone can exhibit varying moisture dynamics during drying and that the rate and nature of evaporative loss differs under different environmental conditions. The results identify the importance of material-environment interactions during drying and that stone micro-environmental conditions cannot be inferred from ambient data alone.
These data have significance for understanding the spatial distribution of stone surface weathering-related morphologies in both the natural and built environments where mineralogical and/or structural heterogeneity creates differences in moisture flux and hence variable drying rates. Such differences may provide a clearer explanation for the initiation and subsequent development of complex weathering responses where areas of significant deterioration can be found alongside areas that exhibit little or no evidence surface breakdown.

Alternative drying techniques for the manufacturing of hydrogel-forming and dissolving microneedle arrays.

Dissolving polymeric microneedle arrays and hydrogel-forming microneedle arrays have attracted much attention during recent years due mainly to their biocompatibility and capacity for enhanced drug delivery. Nevertheless, for the production of this type of devices, typically, a drying step is required. Microneedles are prepared following a micromoulding technique using aqueous blends of Gantrez® S-97. Currently, production of microneedles arrays involves a long drying process of 48 hours. Therefore alternative drying methods were investigated including microwave radiation and hot air convection.

Optimization of osmotic dehydration of papaya followed by air-drying

FERNANDES, Fabiano A. N. et al. Optimization of Osmotic Dehydration of Papaya of followed by air-drying. Food Research Internation, v. 39, p. 492-498, 2006.

Design and development of a diagonal-airflow batch dryer for spatial drying homogeneity

Das Verfahren der Lebensmitteltrocknung wird häufig angewendet, um ein Produkt für längere Zeit haltbar zu machen. Obst und Gemüse sind aufgrund ihres hohen Wassergehalts leicht verderblich durch biochemische Vorgänge innerhalb des Produktes, nicht sachgemäße Lagerung und unzureichende Transportmöglichkeiten. Um solche Verluste zu vermeiden wird die direkte Trocknung eingesetzt, welche die älteste Methode zum langfristigen haltbarmachen ist. Diese Methode ist jedoch veraltet und kann den heutigen Herausforderungen nicht gerecht werden. In der vorliegenden Arbeit wurde ein neuer Chargentrockner, mit diagonalem Luftstömungskanal entlang der Länge des Trocknungsraumes und ohne Leitbleche entwickelt. Neben dem unbestreitbaren Nutzen der Verwendung von Leitblechen, erhöhen diese jedoch die Konstruktionskosten und führen auch zu einer Erhöhung des Druckverlustes. Dadurch wird im Trocknungsprozess mehr Energie verbraucht. Um eine räumlich gleichmäßige Trocknung ohne Leitbleche zu erreichen, wurden die Lebensmittelbehälter diagonal entlang der Länge des Trockners platziert. Das vorrangige Ziel des diagonalen Kanals war, die einströmende, warme Luft gleichmäßig auf das gesamte Produkt auszurichten. Die Simulation des Luftstroms wurde mit ANSYS-Fluent in der ANSYS Workbench Plattform durchgeführt. Zwei verschiedene Geometrien der Trocknungskammer, diagonal und nicht diagonal, wurden modelliert und die Ergebnisse für eine gleichmäßige Luftverteilung aus dem diagonalen Luftströmungsdesign erhalten. Es wurde eine Reihe von Experimenten durchgeführt, um das Design zu bewerten. Kartoffelscheiben dienten als Trocknungsgut. Die statistischen Ergebnisse zeigen einen guten Korrelationskoeffizienten für die Luftstromverteilung (87,09%) zwischen dem durchschnittlich vorhergesagten und der durchschnittlichen gemessenen Strömungsgeschwindigkeit. Um den Effekt der gleichmäßigen Luftverteilung auf die Veränderung der Qualität zu bewerten, wurde die Farbe des Produktes, entlang der gesamten Länge der Trocknungskammer kontaktfrei im on-line-Verfahren bestimmt. Zu diesem Zweck wurde eine Imaging-Box, bestehend aus Kamera und Beleuchtung entwickelt. Räumliche Unterschiede dieses Qualitätsparameters wurden als Kriterium gewählt, um die gleichmäßige Trocknungsqualität in der Trocknungskammer zu bewerten. Entscheidend beim Lebensmittel-Chargentrockner ist sein Energieverbrauch. Dafür wurden thermodynamische Analysen des Trockners durchgeführt. Die Energieeffizienz des Systems wurde unter den gewählten Trocknungsbedingungen mit 50,16% kalkuliert. Die durchschnittlich genutzten Energie in Form von Elektrizität zur Herstellung von 1kg getrockneter Kartoffeln wurde mit weniger als 16,24 MJ/kg und weniger als 4,78 MJ/kg Wasser zum verdampfen bei einer sehr hohen Temperatur von jeweils 65°C und Scheibendicken von 5mm kalkuliert. Die Energie- und Exergieanalysen für diagonale Chargentrockner wurden zudem mit denen anderer Chargentrockner verglichen. Die Auswahl von Trocknungstemperatur, Massenflussrate der Trocknungsluft, Trocknerkapazität und Heiztyp sind die wichtigen Parameter zur Bewertung der genutzten Energie von Chargentrocknern. Die Entwicklung des diagonalen Chargentrockners ist eine nützliche und effektive Möglichkeit um dei Trocknungshomogenität zu erhöhen. Das Design erlaubt es, das gesamte Produkt in der Trocknungskammer gleichmäßigen Luftverhältnissen auszusetzen, statt die Luft von einer Horde zur nächsten zu leiten.

Drying kinetics, biochemical and functional properties of products in convective drying of anchovy (Engraulis Anchoita) fillets

The aim of this work was to study the convective drying of anchovy (Engraulis anchoita) fillets and to evaluate the final product characteristics through its biochemical and functional properties. The drying temperatures were of 50, 60 and 70°C, and the fillet samples were dried with the skins down (with air flow one or the two sides) and skins up (with air flow one side). The drying experimental data were analyzed by Henderson–Pabis model, which showed a good fit (R2 > 0.99 and REQM < 0.05). The moisture effective diffusivity values ranged from 4.1 10–10 to 8.6 10–10 m2 s−1 with the skin down and 2.2 10–10 to 5.5 10–10 m2 s−1 with the skin up, and the activation energy values were 32.2 and 38.4 kJ mol−1, respectively. The product characteristics were significantly affected (p < 0.05) by drying operation conditions. The lower change was in drying at 60°C with air flow for two sides of the samples and skin up. In this condition, the product showed solubility 22.3%; in vitro digestibility 87.4%; contents of available lysine and methionine 7.21 and 2.64 g 100 g−1, respectively; TBA value 1.16 mgMDA kg−1; specific antioxidant activity was 1.91 mMDPPH g−1 min−1, and variation total color was 10.72.