A fully automated spectrophotometric method based on flow-batch analysis has been developed for the determination of clenbuterol including an on-line solid phase extraction using a molecularly imprinted polymer (MIP) as the sorbent. The molecularly imprinted solid phase extraction (MISPE) procedure allowed analyte extraction from complex matrices at low concentration levels and with high selectivity towards the analyte. The MISPE procedure was performed using a commercial MIP cartridge that was introduced into a guard column holder and integrated in the analyzer system. Optimized parameters included the volume of the sample, the type and volume of the conditioning and washing solutions, and the type and volume of the eluent. Quantification of clenbuterol was carried out by spectrophotometry after in-system post-elution analyte derivatization based on azo-coupling using N- (1-Naphthyl) ethylenediamine as the coupling agent to yield a red-colored compound with maximum absorbance at 500nm. Both the chromogenic reaction and spectrophotometric detection were performed in a lab-made flow-batch mixing chamber that replaced the cuvette holder of the spectrophotometer. The calibration curve was linear in the 0.075-0.500mgL-1 range with a correlation coefficient of 0.998. The precision of the proposed method was evaluated in terms of the relative standard deviation obtaining 1.1% and 3.0% for intra-day precision and inter-day precision, respectively. The detection limit was 0.021mgL-1 and the sample throughput for the entire process was 3.4h-1. The proposed method was applied for the determination of CLB in human urine and milk substitute samples obtaining recoveries values within a range of 94.0-100.0%.
- MeSH
- Urinalysis methods MeSH
- Color MeSH
- Clenbuterol analysis isolation & purification urine MeSH
- Colorimetry MeSH
- Humans MeSH
- Limit of Detection MeSH
- Analytic Sample Preparation Methods MeSH
- Molecular Imprinting * MeSH
- Milk Substitutes chemistry MeSH
- Polymers classification MeSH
- Solvents chemistry MeSH
- Temperature MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
This work presents the development of a fully automated flow-batch analysis (FBA) system as a new approach for on-line preconcentration, photodegradation and fluorescence detection in a lab-constructed mixing chamber that was designed to perform these processes without sample dispersion. The system positions the mixing chamber into the detection system and varies the instrumental parameters according to the required photodegradation conditions. The developed FBA system is simple and easily coupled with any sample pretreatment without altering the configuration. This FBA system was implemented to photodegrade and determine the fluorescence of the degradation products of metsulfuron methyl (MSM), a naturally non-fluorescent herbicide of the sulfonylurea׳s family. An on-line solid phase extraction (SPE) and clean up procedure using a C18 minicolumn was coupled to the photodegradation-detection mixing chamber (PDMC) that was located in the spectrofluorometer. An enrichment factor of 27 was achieved. Photodegradation conditions have been optimized by considering the influence of the elution solvent on both the formation of the photoproduct and on the fluorescence signal. Under optimal conditions, the calibration for the MSM determination was linear over the range of 1.00-7.20 µg L(-1). The limit of detection (LOD) was 0.28 µg L(-1); the relative standard deviation was 2.0% and the sample throughput for the entire process was 3h(-1). The proposed method was applied to real water samples from the Bahía Blanca׳s agricultural region (Bahía Blanca, Buenos Aires, Argentina). This method obtained satisfactory recoveries with a range of 94.7-109.8%.
- MeSH
- Arylsulfonates analysis MeSH
- Solid Phase Extraction MeSH
- Spectrometry, Fluorescence MeSH
- Photochemistry methods MeSH
- Photolysis MeSH
- Herbicides analysis MeSH
- Calibration MeSH
- Hydrogen-Ion Concentration MeSH
- Water Pollutants analysis MeSH
- Limit of Detection MeSH
- Reproducibility of Results MeSH
- Sulfonylurea Compounds analysis MeSH
- Light MeSH
- Green Chemistry Technology MeSH
- Water chemistry MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Magnetic resonance spectroscopic imaging (MRSI) involves a huge number of spectra to be processed and analyzed. Several tools enabling MRSI data processing have been developed and widely used. However, the processing programs primarily focus on sophisticated spectra processing and offer limited support for the analysis of the calculated spectroscopic maps. In this paper the jSIPRO (java Spectroscopic Imaging PROcessing) program is presented, which is a java-based graphical interface enabling post-processing, viewing, analysis and result reporting of MRSI data. Interactive graphical processing as well as protocol controlled batch processing are available in jSIPRO. jSIPRO does not contain a built-in fitting program. Instead, it makes use of fitting programs from third parties and manages the data flows. Currently, automatic spectra processing using LCModel, TARQUIN and jMRUI programs are supported. Concentration and error values, fitted spectra, metabolite images and various parametric maps can be viewed for each calculated dataset. Metabolite images can be exported in the DICOM format either for archiving purposes or for the use in neurosurgery navigation systems.
- MeSH
- Electronic Data Processing statistics & numerical data MeSH
- Fourier Analysis MeSH
- Functional Neuroimaging statistics & numerical data MeSH
- Humans MeSH
- Magnetic Resonance Imaging statistics & numerical data MeSH
- Brain metabolism pathology MeSH
- Programming Languages MeSH
- Software * MeSH
- Imaging, Three-Dimensional MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
SUMMARY: ShinySOM offers a user-friendly interface for reproducible, high-throughput analysis of high-dimensional flow and mass cytometry data guided by self-organizing maps. The software implements a FlowSOM-style workflow, with improvements in performance, visualizations and data dissection possibilities. The outputs of the analysis include precise statistical information about the dissected samples, and R-compatible metadata useful for the batch processing of large sample volumes. AVAILABILITY AND IMPLEMENTATION: ShinySOM is free and open-source, available online at gitlab.com/exaexa/ShinySOM. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.
- MeSH
- Algorithms * MeSH
- Metadata MeSH
- Workflow MeSH
- Software * MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
The aim of this systematic study was to analyze the granulometric and rheological behavior of tableting mixtures in relation to tabletability by single tablet and lab-scale batch compression with an eccentric tablet machine. Three mixtures containing 33, 50, and 66% of the cohesive drug paracetamol were prepared. The high compressibility of the powder mixtures caused problems with overcompaction or lamination in the single tablet compression method; due to jamming of the material during the filling of the die, the lab-scale batch compression was impossible. Using high shear granulation, the flow properties and tabletability were adjusted. A linear relationship between the span of granules and the specific energy measured by FT4 powder rheometer was detected. In parallel, a linear relationship between conditioned bulk density and the tensile strength of the tablets at lab-scale batch tableting was noted. The combination of dynamic image analysis and powder rheometry was useful for predicting the tabletability of pharmaceutical mixtures during the single tablet (design) compression and the lab-scale batch compression.
- MeSH
- Acetaminophen * MeSH
- Tensile Strength MeSH
- Powders MeSH
- Drug Compounding MeSH
- Rheology MeSH
- Tablets MeSH
- Particle Size MeSH
- Publication type
- Journal Article MeSH
BACKGROUND: Waste from public green areas represents large quantities of grassy phytomass. The grass is usually utilised by composting, combustion or anaerobic fermentation. However, the classical composts are time-demanding, the quality of accelerated composts is low, combustion is under increasing criticism and conventional anaerobic fermentation requires high investment. RESULTS: A new method of two-fraction anaerobic fermentation of grass waste consisting from a hot maceration, up-flow anaerobic sludge blanket reactor, steam explosion, horizontally stirred batch anaerobic fermentors and a charcoal kiln, all run on waste heat from a co-generation unit was investigated on a commercial scale. CONCLUSION: The results shows that due to faster energy utilisation the two-fraction technology requires smaller fermentors and hence the technology is approximately one-third less costly than conventional systems, with 4 years shorter payback time. Additionally, huge amounts of charcoal are produced. However, the process control and optimisation is more demanding.
- MeSH
- Anaerobiosis MeSH
- Charcoal * MeSH
- Fermentation MeSH
- Poaceae * MeSH
- Plant Leaves * MeSH
- Costs and Cost Analysis MeSH
- Waste Disposal, Fluid methods MeSH
- Refuse Disposal methods MeSH
- Soil * MeSH
- Technology MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
A novel flow-programming setup based on the sequential injection principle is herein proposed for on-line monitoring of temporal events in cell permeation studies. The permeation unit consists of a Franz cell with its basolateral compartment mixed under mechanical agitation and thermostated at 37 °C. The apical compartment is replaced by commercially available Transwell inserts with a precultivated cell monolayer. The transport of drug substances across epithelial cells genetically modified with the P-glycoprotein membrane transporter (MDCKII-MDR1) is monitored on-line using rhodamine 123 as a fluorescent marker. The permeation kinetics of the marker is obtained in a fully automated mode by sampling minute volumes of solution from the basolateral compartment in short intervals (10 min) up to 4 h. The effect of a P-glycoprotein transporter inhibitor, verapamil as a model drug, on the efficiency of the marker transport across the cell monolayer is thoroughly investigated. The analytical features of the proposed flow method for cell permeation studies in real time are critically compared against conventional batch-wise procedures and microfluidic devices.
- MeSH
- Automation methods MeSH
- Biological Transport MeSH
- Epithelial Cells chemistry metabolism MeSH
- Kinetics MeSH
- Humans MeSH
- ATP Binding Cassette Transporter, Subfamily B, Member 1 metabolism MeSH
- Flow Injection Analysis instrumentation methods MeSH
- Rhodamine 123 chemistry metabolism MeSH
- Verapamil chemistry metabolism MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Evaluation Study MeSH
- Research Support, Non-U.S. Gov't MeSH
Mammalian cell perfusion cultures are gaining renewed interest as an alternative to traditional fed-batch processes for the production of therapeutic proteins, such as monoclonal antibodies (mAb). The steady state operation at high viable cell density allows the continuous delivery of antibody product with increased space-time yield and reduced in-process variability of critical product quality attributes (CQA). In particular, the production of a confined mAb N-linked glycosylation pattern has the potential to increase therapeutic efficacy and bioactivity. In this study, we show that accurate control of flow rates, media composition and cell density of a Chinese hamster ovary (CHO) cell perfusion bioreactor allowed the production of a constant glycosylation profile for over 20 days. Steady state was reached after an initial transition phase of 6 days required for the stabilization of extra- and intracellular processes. The possibility to modulate the glycosylation profile was further investigated in a Design of Experiment (DoE), at different viable cell density and media supplement concentrations. This strategy was implemented in a sequential screening approach, where various steady states were achieved sequentially during one culture. It was found that, whereas high ammonia levels reached at high viable cell densities (VCD) values inhibited the processing to complex glycan structures, the supplementation of either galactose, or manganese as well as their synergy significantly increased the proportion of complex forms. The obtained experimental data set was used to compare the reliability of a statistical response surface model (RSM) to a mechanistic model of N-linked glycosylation. The latter outperformed the response surface predictions with respect to its capability and reliability in predicting the system behavior (i.e., glycosylation pattern) outside the experimental space covered by the DoE design used for the model parameter estimation. Therefore, we can conclude that the modulation of glycosylation in a sequential steady state approach in combination with mechanistic model represents an efficient and rational strategy to develop continuous processes with desired N-linked glycosylation patterns. Biotechnol. Bioeng. 2017;114: 1978-1990. © 2017 Wiley Periodicals, Inc.
- MeSH
- Equipment Failure Analysis MeSH
- Models, Biological * MeSH
- Bioreactors * MeSH
- CHO Cells MeSH
- Cricetulus MeSH
- Computer-Aided Design MeSH
- Equipment Design MeSH
- Glycosylation MeSH
- Antibodies, Monoclonal isolation & purification metabolism MeSH
- Perfusion instrumentation methods MeSH
- Computer Simulation MeSH
- Polysaccharides metabolism MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
The influence of industrial (pharmaceutical and chemical) wastewater composition on membrane bioreactor (MBR) performance was investigated in a pilot-scale installation. The study focussed on nitrification performance, which was evaluated based on influent and effluent parameters as well as batch nitrification rate tests. The industrial wastewater was pumped into the MBR in a mixture with municipal wastewater at constant flow rate. The loading of the MBR with industrial wastewater was increased stepwise from 0 to 75% share in the mixed influent to study the adaptation of nitrifying bacteria. Stable nitrification performance was observed until the content of industrial wastewater in the influent reached 40%, with effluent values of around 0.56 mg L(-1) NH4-N and 98.3% ammonia removal. Breakdown of nitratation was observed at a 40% industrial wastewater dose and breakdown of nitritation at a 50% dose, respectively. However, after several months of adaptation, both processes recovered. No nitrification was observed when the industrial wastewater share exceeded 50%. Adaptation of nitrifying bacteria in the MBR was also confirmed by results of kinetic tests. The inhibition effect of the concentrated industrial wastewater to the MBR sludge decreased substantially after several months of exposure, while the inhibition of referential activated sludge remained constant.
- MeSH
- Bioreactors * MeSH
- Water Purification * MeSH
- Nitrogen isolation & purification MeSH
- Filtration MeSH
- Membranes, Artificial * MeSH
- Biological Oxygen Demand Analysis MeSH
- Nitrification * MeSH
- Waste Disposal, Fluid MeSH
- Wastewater MeSH
- Sewage chemistry MeSH
- Permeability MeSH
- Pilot Projects MeSH
- Industrial Waste analysis MeSH
- Cities MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Geographicals
- Cities MeSH
This review deals with techniques and methods used in the study of the function and development of microorganisms occurring in soil with emphasis on the contributions of Czech Academician Ivan Málek and his coworkers or fellows (Jiří Macura, František Kunc) to the development of basic techniques used in soil microbiology. Early studies, including batch cultivation and respirometric techniques, as well as later developments of percolation and continuous-flow methods of cultivation of soil microorganisms are discussed. Recent developments in the application of analytical chemistry (HPLC or GC) and of molecular biological techniques to ecological questions that have revolutionized concepts in soil microbiology and microbial ecology are also briefly mentioned, including denaturing gradient gel electrophoresis (DGGE), temperature gradient gel electrophoresis (TGGE), phospholipid fatty acid analysis (PLFA) and others. The shift of soil microbiology from the study of individual microorganisms to entire microbial communities, including nonculturable species, is briefly discussed.
