Characterization and application of new macroporous membrane ion exchangers

A new ready-to-use unit for high-performance membrane chromatography has been characterized. Its dynamic capacity, resolving power and protein recovery were measured at different flow-rates. The binding capacity was 0.5–2 mg/cm² with a 95% recovery at 10 ml/min irrespective of the protein concentration up to 10 mg/ml. For very-high flow-rates (50 and 100 ml/min) the recovery was 90% and 70%. At these flow-rates, the maximum back-pressure was about 0.1 MPa and was independent of the filtration area. By increasing the filtration area, a proportional capacity increase was obtained, indicating an easy scale-up. High flow-rates had only a slight effect on resolution. This new adsorber was able to purify IgM from supernatant of cell culture of a human hybridoma in less than 8 min with a high degree of purity (95%).     

Analytical and preparative applications of magnetic split-flow thin fractionation on several ion-labeled red blood cells

Background

Magnetic Split-flow thin (SPLITT) fractionation is a newly developed technique for separating magnetically susceptible particles. Particles with different field-induced velocities can be separated into two fractions by adjusting applied magnetic forces and flow-rates at inlets and outlets.

Methods

Magnetic particles, Dynabeads, were used to test this new approach of field-induced velocity for susceptibility determination using magnetic SF at different magnetic field intensities. Reference measurements of magnetic susceptibility were made using a superconducting quantum interference device (SQUID) magnetometer. Various ion-labeled red blood cells (RBC) were used to study susceptibility determination and throughput parameters for analytical and preparative applications of magnetic SPLITT fractionation (SF), respectively. Throughputs were studied at different sample concentrations, magnetic field intensities, and channel flow-rates.

Results

The susceptibilities of Dynabeads determined by SPLITT fractionation (SF) were consistent with those of reference measurement using a superconducting quantum interference device (SQUID) magnetometer. Determined susceptibilities of ion-labeled RBC were consistent within 9.6% variations at two magnetic intensities and different flow-rates. The determined susceptibilities differed by 10% from referenced measurements. The minimum difference in magnetic susceptibility required for complete separation was about 5.0 × 10^-6 [cgs]. Sample recoveries were higher than 92%. The throughput of magnetic SF was approximately 1.8 g/h using our experimental setup.

Conclusion

Magnetic SF can provide simple and economical determination of particle susceptibility. This technique also has great potential for cell separation and related analysis. Continuous separations of ion-labeled RBC using magnetic SF were successful over 4 hours. The throughput was increased by 18 folds versus early study. Sample recoveries were 93.1 ± 1.8% in triplicate experiments.

     

Development of a simultaneous bioreactor system for characterization of gas production kinetics of methanogenic archaea at high pressure

Cultivation of methanogens under high pressure offers a great opportunity in biotechnological processes, one of which is the improvement of the gas‐liquid transfer of substrate gases into the medium broth. This article describes a newly developed simultaneous bioreactor system consisting of four identical cultivation vessels suitable for investigation of microbial activity at pressures up to 50 bar and temperatures up to 145°C. Initial pressure studies at 10 and 50 bar of the autotrophic and hydrogenotrophic methanogens Methanothermobacter marburgensis, Methanobacterium palustre, and Methanobacterium thermaggregans were performed to evaluate the reproducibility of the system as well as to test the productivity of these strains. The strains were compared with respect to gas conversion (%), methane evolution rate (MER) (mmol L^‐1 h^?1), turnover rate (h^?1), and maximum conversion rate (k_min) (bar h^?1). A pressure drop that can be explained by the reaction stoichiometry showed that all tested strains were active under pressurized conditions. Our study sheds light on the production kinetics of methanogenic strains under high‐pressure conditions. In addition, the simultaneous bioreactor system is a suitable first step screening system for analyzing the substrate uptake and/or production kinetics of gas conversion and/or gas production processes for barophilic or barotolerant microbes.     

Steady pressure-flow relationship in a cast of the upper and central human airways

Pressure drops across the upper (larynx) and central airways of a human lung cast were measured at steady state inspiratory and expiratory flows. Air, HeO₂ and SF6-O₁ gas mixtures were used at tracheal Reynolds' numbers ranging from 145 to 30 000. The pressure-flow characteristics of the model were analysed using standard pressure-flow diagrams and Moody plots. We found that the asymmetry between inspiratory and expiratory resistances, observed in the central airways (larynx excluded), was markedly reduced in the presence of the larynx. However, static pressure differences were greater across the entire model of the upper and central airways than across the model of the five generations of the tracheo-bronchial tree (without larynx) at the same flow-rates. In addition, our results showed that the presence of the larynx tended to reduce the zone of fully developed laminar flow in the Moody diagram with the higher density gas, while extending the zone of turbulent flow even for the low density gas at low Reynold's numbers.     

Removal of toluene in waste gases using a biological trickling filter

The removal of toluene from waste gas was studied in a trickling biofilter. A high level of water recirculation (4.7 m h^–1) was maintained in order to keep the liquid phase concentration constant and to achieve a high degree of wetting. For loads in the range from 6 to 150 g m^–3 h^–1 the maximum volumetric removal rate (elimination capacity) was 35±10 g m^–3 h^–1, corresponding to a zero order removal rate of 0.11±0.03 g m^–2 h^–1 per unit of nominal surface area. The surface removal was zero order above the liquid phase concentrations of approximately 1.0 g m^–3, corresponding to inlet gas concentrations above 0.7–0.8 g m^–3. Below this concentration the surface removal was roughly of first order. The magnitude of the first order surface removal rate constant, k_1A , was estimated to be 0.08–0.27 m h^–1 (k_1A a=24–86 h^–1). Near-equilibrium conditions existed in the gas effluent, so mass transfer from gas to liquid was obviously relatively fast compared to the biological degradation. An analytical model based on a constant liquid phase concentration through the trickling filter column predicts the effluent gas concentration and the liquid phase concentration for a first and a zero order surface removal. The experimental results were in reasonable agreement with a very simple model valid for conditions with an overall removal governed by the biological degradation and independent of the gas/liquid mass transfer. The overall liquid mass transfer coefficient, K_La, was found to be a factor 6 higher in the system with biofilm compared to the system without. The difference may be explained by: 1. Difference in the wetting of the packing material, 2. Mass transfer occurring directly from the gas phase to the biofilm, and 3. Enlarged contact area between the gas phase and the biofilm due to a rough biofilm surface.     

Gas exchange in San Francisco Bay

Gas exchange across the air-water interface is one of the most important processes controlling the concentrations of dissolved gases in estuarine systems. A brief review of principles and equations to predict gas exchange indicates that both current shear and wind shear are possible sources of turbulence for controlling gas exchange rates in estuaries. Rates of exchange determined by constructing a mass balance for radon-222 indicate that wind shear is dominant in San Francisco Bay. Because many estuaries have wind shear and current speeds comparable to this system, this conclusion may be true for other systems as well. A compilation of gas exchange rates measured in San Francisco Bay with those for other wind-dominated systems updates previous compilations and yields an equation for predicting gas exchange: K _l = 34.6 R _v (D_m20)^1/2 (U₁₀)^1.5 where R is the ratio of the kinematic viscosity of pure water at 20° C to the kinematic viscosity of water at the measured temperature and salinity, D_m20 is the molecular diffusivity of the gas of interest at 20°C in cm² s^–1, U₁₀ is the wind speed at 10 meters above the surface in m s^–1, and K_L is the liquid phase gas transfer coefficient in m d^–1. This relationship fits the available field data within 20% for wind speeds between 3 and 12 m s^–1. It is used to show that the residence time of dissolved oxygen in San Francisco Bay should range from 2 days during windy summer periods to as much as 15 days during calm winter periods. Because these times are short compared to time constants for other processes controlling oxygen distribution in this system, dissolved oxygen concentrations in San Francisco Bay are usually near atmospheric equilibrium. Other systems, such as Chesapeake Bay, may differ. There, despite ample air-water gas exchange rates, some bottom waters become anoxic during summer months due to slow vertical mixing.     

Acetone biodegradation in a trickle-bed biofilter

Laboratory scale-studies on acetone biodegradation in a trickle-bed biofilter were carried out using using two identically sized columns, one of which was packed with coconut fibre and the other with plexiglass chips. The columns were inoculated with two strains of bacteria: Burkholderia cepacia (syn. Pseudomonas cepacia) and Acinetobacter baumannii. The continuous process of air purification was carried out at incremental acetone concentrations from 0.3 to 2.5 g m⁻³ air and air flow-rates from 0.1 to 0.3 m³ h⁻¹. A maximum acetone elimination capacity of 95.8 g m⁻³ filter bed h⁻¹ was achieved at air flow-rate of 36 m h⁻¹.     

Tradeoffs between metabolic rate and spiracular conductance in discontinuous gas exchange of Samia cynthia (Lepidoptera, Saturniidae)

The insect tracheal system is a unique respiratory system, designed for maximum oxygen delivery at high metabolic demands, e.g. during activity and at high ambient temperatures. Therefore, large safety margins are required for tracheal and spiracular conductance. Spiracles are the entry to the tracheal system and play an important role in controlling discontinuous gas exchange (DGC) between tracheal system and atmosphere in moth pupae. We investigated the effect of modulated metabolic rate (by changing ambient temperature) and modulated spiracular conductance (by blocking all except one spiracles) on gas exchange patterns in Samia pupae. Both, spiracle blocking and metabolic rates, affected respiratory behavior in Samia cynthia pupae. While animals showed discontinuous gas exchange cycles at lower temperatures with unblocked spiracles, the respiratory patterns were cyclic at higher temperatures, with partly blocked spiracles or a combination of these two factors. The threshold for the transition from a discontinuous (DGC) to a cyclic gas exchange (^cycGE) was significantly higher in animals with unblocked spiracles (18.7 nmol g⁻¹ min⁻¹ vs. 7.9 nmol g⁻¹ min⁻¹). These findings indicate an important influence of spiracle conductance on the DGC, which may occur mostly in insects showing high spiracular conductances and low metabolic rates.     

A microbiological study of an underground gas storage in the process of gas injection

The liquid phase of different units of an underground gas storage (UGS) in the period of gas injection was studied with respect to its hydrochemical composition and characterized microbiologically. The presence of viable aerobic and anaerobic bacteria was revealed in the UGS stratal and associated waters. An important source of microorganisms and biogenic elements in the ecosystem studied is water and various technogenic admixtures contained in trace amounts in the gas entering from the gas main in the period of gas injection into the storage. Owing to this fact, the bacterial functional diversity, number, and activity are maximal in the system of gas treatment and purification and considerably lower in the observation well zone. At the terminal stages, the anaerobic transformation of organic matter in the UGS aqueous media occurs via sulfate reduction and methanogenesis; exceptionally high rates of these processes (up to 4.9 × 10⁵ ng S^2? l^?1 day^?1 and 2.8 × 10⁶ nl CH₄ l^?1 day^?1, respectively) were recorded for above-ground technological equipment.     

An automated continuous determination of oxygen with high sensitivity

A sensitive, rapid, and precise method is described for the continuous determination of oxygen in gases. The principle of the method is based on the reaction of O₂ with an alkaline catechol or pyrogallol solution, which is combined with a Fe²⁺ solution to increase the sensitivity of the color reaction. The development of the color takes place in a tube system provided with a proportioning pump and is read automatically on a recorder after passing a flow cell of a photometer. The lower limit of sensitivity of this method is 0.05 μl of O₂ min^?1. Thus, in a gas flow of ≈0.5 ml min^?1, an oxygen concentration of ≥0.01% (v/v) can be determined. If the gas flow is increased to ≈2.5 ml min^?1, this limit of sensitivity is lowered to ≥0.002% (v/v). Since a 2-min period is necessary for the measurement, the volume of the sample has to be 1 ml in the first case and 5 ml in the second.     

Measurement of breath-by-breath gas exchange during general anaesthesia using a single pneumotachograph

A breath-by-breath gas exchange measurement system using a single pneumotachograph suitable for use during general anaesthesia is described. The system's accuracy has been assessed by a combination of error sensitivity analysis, laboratory testing of the component measurements used to calculate gas exchange and measurements on volunteers and patients. The system is shown to have a mean accuracy of ± 2.6 ml breath⁻¹ for V_CO2 measurements, ± 7.12 ml breath⁻¹ for V_O2 and ± 5.55 ml breath⁻¹ for V_N2O measurement. The application of a lung gas stores correction using argon improved between breath variability by 50%.     

High-sensitivity phenylthiohydantoin amino acid analysis using conventional and microbore chromatography

Reversed-phase microbore high-performance liquid chromatography was investigated for high-sensitivity analysis of phenylthiohydantoin (PTH) amino acids. A mixed nitrile alkylsilane bonded phase was developed and ternary gradient elution conditions were devised for resolution 150 × 4.6 mm I.D. column and transferred to a 150 × 1 mmI.D. microbore column. The performance of these columns was evaluated in terms of PTH amino acid resolution, enhanced sample detectability, and retention time precision. For this work a general purpose high-performance liquid chromatograph was modified to reduce extra column band broadening and a preformed gradient elution technique was developed to achieve rapid analysis times at microbore flow-rates. The microbore high-performance liquid chromatographic system is useful for high-sensitivity analysis of PTH amino acids in micro-sequencing applications.     

Optimize Flue Gas Settings to Promote Microalgae Growth in Photobioreactors via Computer Simulations

Flue gas from power plants can promote algal cultivation and reduce greenhouse gas emissions¹. Microalgae not only capture solar energy more efficiently than plants³, but also synthesize advanced biofuels^2-4. Generally, atmospheric CO₂ is not a sufficient source for supporting maximal algal growth⁵. On the other hand, the high concentrations of CO₂ in industrial exhaust gases have adverse effects on algal physiology. Consequently, both cultivation conditions (such as nutrients and light) and the control of the flue gas flow into the photo-bioreactors are important to develop an efficient “flue gas to algae” system. Researchers have proposed different photobioreactor configurations^4,6 and cultivation strategies^7,8 with flue gas. Here, we present a protocol that demonstrates how to use models to predict the microalgal growth in response to flue gas settings. We perform both experimental illustration and model simulations to determine the favorable conditions for algal growth with flue gas. We develop a Monod-based model coupled with mass transfer and light intensity equations to simulate the microalgal growth in a homogenous photo-bioreactor. The model simulation compares algal growth and flue gas consumptions under different flue-gas settings. The model illustrates: 1) how algal growth is influenced by different volumetric mass transfer coefficients of CO₂; 2) how we can find optimal CO₂ concentration for algal growth via the dynamic optimization approach (DOA); 3) how we can design a rectangular on-off flue gas pulse to promote algal biomass growth and to reduce the usage of flue gas. On the experimental side, we present a protocol for growing Chlorella under the flue gas (generated by natural gas combustion). The experimental results qualitatively validate the model predictions that the high frequency flue gas pulses can significantly improve algal cultivation.     

Batch culture of Alcaligenes eutrophus ATCC 17697T using recycled gas closed circuit culture system

Batch culture of Alcaligenes eutrophus ATCC 17697^T using the recycled gas closed circuit culture system was done with the intention of developing a practical fermentation system for industrial culture of autotrophs. The gas phase of the culture system consisted of substrate gas so that gases in this culture system could be recycled forever as long as the amount of the gas consumed would be refilled. All gasses supplied into this system could be completely used without any loss as exhaust. Thus, this system assures high gas usage efficiency as well as operation safety. Studies on the effect of oxygen concentration showed the high oxygen concentration in substrate gas mixture suppressed the specific growth rate while a low oxygen concentration promoted the growth rate. This introduced the possibility of development of an explosion-free fermentation system with a high growth rate if appropriate system design and fermentation conditions were given. Since this system is closed, material balance and elementary analysis provide the ultimately accurate stoichiometry of the autotrophic culture of this bacterium.     

Biological kinetics evaluation of anaerobic stabilization pond treatment of palm oil mill effluent

Biological kinetic (bio-kinetic) study of the anaerobic stabilization pond treatment of palm oil mill effluent (POME) was carried out in a laboratory anaerobic bench scale reactor (ABSR). The reactor was operated at different feed flow-rates of 0.63, 0.76, 0.95, 1.27, 1.9 and 3.8 l of raw POME for a day. Chemical oxygen demand (COD) as influent substrates was selected for bio-kinetic study. The investigation showed that the growth yield (Y_G), specific biomass decay (b), maximum specific biomass growth rate (μ_max), saturation constant (K_s) and critical retention time (Θ_c) were in the range of 0.990 g VSS/g COD_removed day, 0.024 day⁻¹, 0.524 day⁻¹, 203.433 g COD l⁻¹ and 1.908 day, respectively.     

Treatment of high-concentration gaseous benzene streams using a novel bioreactor system

A continuous treatment system combining a packed-bed column and a two-phase partitioning bioreactor has been designed to treat high-concentration benzene-containing gas streams. 1-Octadecene was used in a closed loop as an absorbant to scrub benzene in the counter-current column, after which it was transferred to the two-phase partitioning bioreactor to partition benzene into the 1 l aqueous phase for degradation by Klebsiella sp. The solvent was then recirculated back to the absorber. A gas stream containing 20 mg l^–1 benzene at a flow rate of 60 l h^–1 was introduced to the system, and the benzene was degraded at a biological removal efficiency of 87% at steady state.     

Mass Transfer from Gas Bubbles to Impinging Flow of Biological Fluids with Chemical Reaction

The rates of mass transfer from a gas bubble to an impinging flow of a biological fluid such as whole blood and plasma are investigated analytically and experimentally. Gases commonly found dissolved in body fluids are included. Consideration is given to the effects of the chemical reaction between the dissolved gas and the liquid on the rate of mass transfer. Through the application of boundary layer theory the over-all transfer is found to be Sh/(Re)^1/2 = 0.845 Sc^1/3 in the absence of chemical reaction, and Sh/(Re) ^1/2 = F′ (0) in the presence of chemical reaction, where Sh, Re, and Sc are the Sherwood, Reynolds, and Schmidt numbers, respectively, and F′ (0) is a function of Sc and the dimensionless reaction rate constant. Analytical results are also obtained for the bubble lifetime and the bubble radius-time history. These results, which are not incompatible with experimental results, can be applied to predict the dissolution of the entrapped gas emboli in the circulatory system of the human body.     

High cell density suspension culture of mammalian anchorage independent cells: Oxygen transfer by gas sparging and defoaming with a hydrophobic net

Gas sparging directly into the culture-broth is not done in cell culture, except when the gas flow rate is very small, because much foaming occurs.During screening of defoaming methods, foam was observed to be broken up effectively when it made contact with a net fabricated from hydrophobic materials. Providing a highly efficient oxygen supply to suspension culture was tried using the new defoaming method. In a 5 1 reactor equipped with the foam-eliminating net fabricated with polysiloxane, oxygen was transferred at 21 mmole/l·h equivalent to an about forty-fold higher rate than in conventional surface aeration. This was equivalent to a consumption rate of 1×10⁸ cells/ml, even at a low oxygen gas flow rate of 0.1 cm/s corresponding to a fourth of the gas flow rate when foam leaked through the net.Perfusion culture of rat ascites hepatoma cell JTC-1 was successfully carried out in the 51 scale culture system with the net and a hydrophobic membrane for cell filtration. The viable cell concentration reached 2.7×10⁷ cells/ml after twenty-seven days, in spite of the nutrient-deficient condition of the lower medium exchange rate, that is, a working volume a day, and viability was maintained at more than 90%. In a 1.21 scale culture of mouse-mouse hybridoma cell STK-1, viable cell concentration reached 4×10⁷ cells/ml. These results showed that oxygen transfer by gas sparging with defoaming was useful for high density suspension culture. A foam-breaking mechanism was proposed.Abbreviations Eagle's MEM Eagle's minimal essential medium - Dulbecco's modified Eagle MEM Dulbecco's modified Eagle minimal essential medium     

Acetylcholine turnover estimation in brain by gas chromatography-mass spectrometry

A gas chromatograph/quadrupole mass spectrometer system has been employed to estimate the turnover of acetylcholine in mouse brain $\text{in vivo}$ following a pulse intravenous injection of choline, using discrete deuterium labelled variants of choline and acetylcholine as tracer and internal standards. There appear to be two functional pools with turnover rates of 1.4 and 7.9 nmol gm^?1min^?1.     

Scale study of direct synthesis of dimethyl ether from biomass synthesis gas

We investigated the synthesis of dimethyl ether (DME) from biomass synthesis gas using a kind of hybrid catalyst consisting of methanol and HZSM-5 zeolite in a fixed-bed reactor in a 100 ton/year pilot plant. The biomass synthesis gas was produced by oxygen-rich gasification of corn core in a two-stage fixed bed. The results showed that CO conversions reached 82.00% and 73.55%, the selectivities for DME were 73.95% and 69.73%, and the space–time yields were 124.28 kg m^− 3 h^− 1 and 203.80 kg m^− 3 h^− 1 when gas hourly space velocities were 650 h^− 1 and 1200 h^− 1, respectively. Deoxidation and tar removal from biomass synthesis gas was critical to the stable operation of the DME synthesis system. Using single-pass synthesis, the H₂/CO ratio improved from 0.98–1.17 to 2.12–2.22. The yield of DME would be increased greatly if the exhaust was reused after removal of the CO₂.