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Stability of elliptical cylinders in two-dimensional channel flow.

The flow around rigid cylinders of elliptical cross section placed transverse to Poiseuille flow between parallel plates was simulated to investigate issues related to the tumbling of red blood cells and other particles of moderate aspect ratio in the similar flow in a Field Flow Fractionation (FFF) channel. The torque and transverse force on the cylinder were calculated with the cylinder freely translating, but prevented from rotating, in the flow. The aspect ratios (long axis to short axis) of the elliptical cylinders were 2, 3, 4, and 5. The cylinder was placed transversely at locations of y0/H = 0.1, 0.2, 0.3, and 0.4, where y0 is the distance from the bottom of the channel and H is the height of the channel, and the orientation of the cylinder was varied from 0 to 10 deg with respect to the axis of the channel for a channel Reynolds number of 20. The results showed that equilibrium orientations (indicated by a zero net torque on the cylinder) were possible for high-aspect-ratio cylinders at transverse locations y0/H < 0.2. Otherwise, the net torque on the cylinder was positive, indicating that the cylinder would rotate. For cylinders with a stable orientation, however, a transverse lift forced existed up to about y0/H = 0.25. Thus, a cylinder of neutral or low buoyancy might be lifted with a stable orientation from an initial position near the wall until it reached y0/H < 0.2, whereupon it would begin to tumble or oscillate. The dependence of lift and torque on cylinder orientation suggested that neutral or low-buoyancy cylinders may oscillate in both transverse location and angular velocity. Cylinders more dense than the carrier fluid could be in equilibrium both in terms of orientation and transverse location if their sedimentation force matched their lift force for a location y0/H < 0.2.

Cell Movement↗

A survey of separative techniques.

Separative science has recently undergone numerous advances. This article discusses many developments and trends in liquid, ion, gas, and countercurrent chromatography, field-flow fractionation, and electrophoresis.

Chromatography, Gas↗

Rifampicin carrying polyhydroxybutyrate microspheres as a potential chemoembolization agent.

In this study, we attempted to prepare microspheres from a microbial biodegradable polyester, i.e. polyhydroxybutyrate (PHB) as a potential chemoembolization agent. The drug loaded PHB microspheres were prepared by a solvent evaporation technique, in which methylene chloride, distilled water, and polyvinyl alcohol were utilized as the solvent, dispersion medium, and emulsifier, respectively. Microspheres were obtained within a size range of 5-100 microns by changing the initial polymer/solvent ratio, emulsifier concentration, stirring rate, and initial drug concentration. It was possible to obtain PHB with very narrow size distributions by applying gravity field-flow fractionation technique. Very high drug loadings of up to 407.6 mg rifampicin/g PHB were achieved. Drug release rates were very rapid. Almost 90% of the drug loaded was released in about 24 h. Both the size and drug content of PHB microspheres were found to be effective in controlling the drug release from these microspheres.

Antibiotics, Antitubercular↗

[Historical study of separation technology innovation in the field of pharmaceutical science].

The separation of organic compounds has been carried out using the method of distillation and recrystallization since the 17th century.1st separation technology innovation (1950s to 1960s). After World War II, the science of separating organic compounds evolved to the use of paper or column chromatography. 2nd separation technology innovation (1960s to 1980s). In the 1960s, thin-layer chromatograpy (TLC) was developed. TLC is a convenient analytical technique for organic compounds. Separation methods for organic compounds using column chromatography and preparative TLC were carried out as practical applications. Gas chromatography (GC) was also introduced in the 1960s. GC is a useful analytical method for a mixture of various volatile organic compounds. Many GC instruments with new kinds of sensors were developed. Gas chromatography of nonbenzenoid organometallic compounds and metal complexes was successfully conducted by the author. 3rd separation technology innovation (1980s to 1990s). In the 1980s, high-performance liquid-chromatography (HPLC) was developed. HPLC analytical methods were useful for various kinds of organic compounds. In the 1990s, LC-MS and LC-NMR instruments were introduced for biomedical products. 4th separation technology innovation (1990s to present). Several new ionization methods for mass-spectrometry (electrospray, matrix-aided lazer desorption ionization, etc) are applied for proteins and biomedical products. Several new separation technologies (supercritical field chromatography, capillary electrophoresis, field-flow fractionization, etc.) are now being developed.

History, 20th Century↗

Scientific achievements of Jack Kirkland to the development of HPLC and in particular to HPLC silica packings--a personal perspective.

Joseph (Jack) Kirkland is one of the outstanding protagonists of modern column liquid chromatography (HPLC). He started in 1953 as an analytical chemist at the Experimental Station of Du Pont de Nemours & Co, Wilmington, Delaware, analyzing pesticides by gas chromatography (GC). He early recognized the potential of HPLC as a powerful separation technique at the end of 1960. His major contributions to HPLC were in the field of silica based packings and stationary phases. At the beginning of the 1970's he manufactured Porous Layer Beads and later microparticulate porous silicas based at the Zorbax technology. He made outstanding contributions in the field of instrument design for HPLC and in the field of Sedimentation Field Flow Fractionation (SFFF), in HPLC method development and optimization strategies. In 1992 Jack resigned from Du Pont de Nemours & Co, Wilmington, Delaware, and joined Rockland Technologies, Newport, Delaware, as a Director of Research and Development. During this period he successfully developed a series of novel, designed reversed phase silicas. He resigned from Rockland technologies, now Agilent Technologies, Newport, Delaware, in 2001, but always remained dedicated to HPLC.

Chromatography, High Pressure Liquid↗

Characterization of polyethyleneglycol-stabilized, manganese-substituted hydroxylapatite (MnHA-PEG). A potential MR blood pool agent.

PURPOSE: To optimize the performance (or efficacy) of a potential particulate blood pool agent for MR angiography by varying the particle size. The colloidal system under investigation was polyethylene glycol-stabilized manganese-substituted hydroxylapatite (MnHA-PEG). MATERIAL AND METHODS: Several MnHA-PEG formulations were prepared using various length PEGs (MW = 140-2000). Products were characterized in vitro by dynamic light scattering (DLLS), field flow fractionation (FFF), and relaxometry; and in vivo by blood clearance kinetics in rabbits, and by analytical electron microscopy (EM). RESULTS: The particle size distribution (PSD) consisted only of small particles (approximately 10-nm diameter) when approximately 40 mo1% PEG was used. At approximately 20 mo1% PEG, larger particles (approximately 100 nm), which are aggregates of the small ones, were also present. The water proton relaxation profiles of the particles in plasma were different from that of the free Mn2+. In plasma, the large aggregates were broken down into the smaller particles which were stable. Although the small particles were efficient relaxation enhancing agents, they were cleared from the blood approximately 3 times faster than the approximately 100-nm diameter aggregates, probably as a consequence of leakage into the extravascular space. Variation of PEG size had no effect on particle characteristics or on blood clearance. Analytical EM of rabbit liver specimens indicated some retention of Mn in mitochondria at the time point when Mn content of other subcellular structures returned to baseline. CONCLUSION: DLLS and FFF are complementary techniques for sizing particulate MR contrast media. Small MnHA particles are more efficient T1-shortening agents than large ones but they are prone to leakage from the vascular space. Within the MW range explored, the length of PEG molecule had no effect on blood clearance of the MnHA particles. Larger aggregates of MnHA-PEG break down into stable small particles in plasma. Mn clears from the subcellular structures within hepatocytes within 60 min after i.v. MnHA-PEG administration except from the mitochondria in which it appears to accumulate.

Animals↗

Nitric-oxide planar laser-induced fluorescence applied to low-pressure hypersonic flow fields for the imaging of mixture fraction.

The scalar-field imaging of a hypersonic mixing flow is performed in a mixing facility that is shock tunnel driven. The instantaneous mixture-fraction field of a hypersonic two-dimensional mixing layer (M1 = 5.1, M2 = 0.3) is determined with a temperature-insensitive planar laser-induced fluorescence technique with nitric oxide (NO) as the tracer species. Single-shot images are obtained with the broadband excitation of a reduced temperature-sensitivity transition in the A2 sigma+ <-- X2 II(1/2) (0, 0) band of NO near 226 nm. The instantaneous mixture-fraction field at a convective Mach number of 2.64 is shown to be nearly identical to a typical diffusive process, supporting the notion of gradient-transport mixing models for highly compressible mixing layers.

Journal Article↗

Determination of total and EDTA extractable metal distributions in the colloidal fraction of contaminated soils using SdFFF-ICP-HRMS.

Newly developed methods involving an on-line combination of sedimentation field-flow fractionation-inductively coupled plasma-high resolution mass spectrometry (SdFFF-ICP-HRMS) have been used to study the distributions of extractable heavy metals in a soil which had been treated with sewage sludge contaminated with Cu or Pb. The relationship of these metals with other elements in the colloidal fraction was also investigated. The colloidal fraction from the soil was obtained by repeated gravitational sedimentation and extracted with 0.11 M acetic acid, 0.1 M hydroxylamine hydrochloride, 0.05 M ethylenediaminetetraacetic acid disodium salt (EDTA) or aqua regia to assess the potential availability of the metals Cu and Pb. Large proportions of the Cu and Pb were extracted by EDTA, approaching that removed by aqua regia, whereas < 10% of the aqua regia extractable metals were removed by acetic acid and hydroxylamine chloride. The distributions of the heavy metals, the major mineral forming element (Al) and the elements forming sesquioxides (Fe and Mn) within different size classes (0.05-1 microm) of the colloidal fraction were measured using SdFFF-ICP-HRMS before and after extraction with EDTA. This information provides an insight into the composition of the colloids and the distributions of metal contaminants. In the contaminated soil colloids, the concentration of Fe, Mn and Pb is greatest in the smaller particles (<0.2 microm). In contrast, the Cu concentration is constant over the size range studied. Iron oxide surface coatings probably play a significant role in Pb adsorption on soil particles, but may be less important for Cu. The combination of selective chemical extraction, SdFFF and ICP-HRMS provides a means of determining the distribution of potentially available heavy metals within the colloidal fraction of contaminated soils.

Chelating Agents↗

Cell sorting by one gravity SPLITT fractionation.

The need for innovative separative techniques suitable for the fractionation of biomaterials prompted this investigation into the performance of the gravitational split-flow thin channel (G-SPLITT) system as a cell sorter. The rigorous mathematical description of the separation mechanism allows achievement of fast separation of several million myeloma cells from healthy splenocytes using flow conditions calculated from theory. Separation in G-SPLITT is based on differences in sedimentation rate. For accurate prediction of the optimal working conditions, this parameter was directly measured by cell tracking velocimetry rather than relying on a measure of diameter (by Multisizer) and an assumed density for each cell population. We also discuss the influence of different flow conditions on the effectiveness of separation.

Cell Separation↗

Continuous particle separation through deterministic lateral displacement.

We report on a microfluidic particle-separation device that makes use of the asymmetric bifurcation of laminar flow around obstacles. A particle chooses its path deterministically on the basis of its size. All particles of a given size follow equivalent migration paths, leading to high resolution. The microspheres of 0.8, 0.9, and 1.0 micrometers that were used to characterize the device were sorted in 40 seconds with a resolution of approximately 10 nanometers, which was better than the time and resolution of conventional flow techniques. Bacterial artificial chromosomes could be separated in 10 minutes with a resolution of approximately 12%.

Chemical Fractionation↗

Proton magnetic resonance imaging of flow motion of heavy water injected into a hollow fiber dialyzer filled with saline.

Observations using MRI were performed for the motion of heavy water injected into a hollow fiber dialyzer. A cylindrical dialyzer houses a bundle of 10,000 hollow fibers. Because blood components permeate through the hollow fiber membrane from the interior to the exterior of the hollow fiber, which is the dialysate flow path, uniformity of dialysate flow is required. The dialyzer was initially filled with saline and heavy water was injected into the inlet port of the dialysate flow path. MRI tuned for protons could distinguish the injected heavy water from the already present saline. Due to the specific gravity difference, MRI could observe the sedimentation of the injected heavy water flowing beneath the already present saline. The uniformity of the dialysate flow was supported by the finding that the injected heavy water brought about uniform sedimentation and distributed the already present saline uniformly throughout the entire volume of the dialyzer.

Deuterium Oxide↗

Separation of lipids from blood utilizing ultrasonic standing waves in microfluidic channels.

A method to continuously separate different particle types in a suspension is reported. Acoustic forces in a standing wave field were utilized to discriminate lipid particles from erythrocytes in whole blood. The presented technology proposes a new method of cleaning, i.e. removing lipid emboli from, shed blood recovered during cardiac surgery. Blood contaminated with lipid particles enter a laminar flow micro channel. Erythrocytes and lipid particles suspended in blood plasma are exposed to a half wavelength standing wave field orthogonal to the direction of flow as they pass through the channel. Because of differences in compressibility and density the two particle types move in different directions, the erythrocytes towards the centre of the channel and the lipid particles towards the side walls. The end of the channel is split into three outlet channels conducting the erythrocytes to the centre outlet and the lipid particles to the side outlets due to the laminar flow profile. The separation channel was evaluated in vitro using polyamide spheres suspended in water, showing separation efficiencies approaching 100%. The system was also evaluated on whole blood using tritium labelled lipid particles added to bovine blood. More than 80% of the lipid particles could be removed while approximately 70% of the erythrocytes were collected in one third of the original fluid volume. The study showed that the further reduced micro channel dimensions provided improved performance with respect to; (i) separation efficiency, (ii) actuation voltage, and (iii) volumetric throughput as compared to earlier work.

Acoustics↗

Continuous flow microextraction combined with high-performance liquid chromatography for the analysis of pesticides in natural waters.

Continuous flow microextraction (CFME) combined with high-performance liquid chromatography-ultraviolet (HPLC-UV) detection has been applied to the analysis of five widely used pesticides, simazine, fensulfothion, etridiazole, mepronil and bensulide, present at trace levels in water samples. CFME employs a single organic solvent drop positioned at the tip of a polyether ether ketone (PEEK) tubing, which is immersed in a continuous flowing aqueous sample solution in a 0.5-ml glass chamber. The PEEK tubing acts as the organic drop holder and fluid delivery duct. Analytes are partitioned between the organic drop and the bulk sample solution. Important extraction factors including type of solvent, its volume, sample solution flow rate, extraction time, its pH and addition of salt were investigated. All pesticides exhibit good linearity in the investigated concentration range of 25-250 ng ml(-1) with coefficients of determination (R2) ranging from 0.9879 to 0.9999 under the optimized conditions. Detection limits lower than 4 ng ml(-1) were obtained for all analytes. The method was evaluated by analyzing natural water sample collected from a reservoir in Singapore. This study for the first time demonstrated the compatibility of CFME procedure and HPLC separation.

Chromatography, High Pressure Liquid↗

Complexities in ETS-domain transcription factor function and regulation: lessons from the TCF (ternary complex factor) subfamily. The Colworth Medal Lecture.

The ETS-domain transcription factor family can be divided into a series of subfamilies. Elk-1 represents the founding member of the ternary complex factor (TCF) subfamily. By focusing on the TCF subfamily, we can demonstrate the complexities that exist in the function and regulation of ETS-domain transcription factors. This article focuses on Elk-1 in detail and summarizes the functions of other TCFs. The key themes covered include the domain structure of the TCFs, the mechanisms of complex formation with serum response factor, regulation of TCFs by mitogen-activated protein kinase cascades, and transcriptional regulatory properties of the TCFs. Finally, the emerging role of the TCFs in vivo is discussed. A picture is developing indicating that, while these proteins exhibit significant sequence and functional conservation, key differences in their structure and regulation are being identified which may relate to unique functions of these proteins in vivo.

Amino Acid Sequence↗

Cell cytometry with a light touch: sorting microscopic matter with an optical lattice.

Lab-on-a-chip design is a key technology for increasing both the reliability and the functionality of many different preparation and diagnostic techniques in biomedicine. The drive towards ever more integrated lab-on-a-chip designs requires increasingly complex microfluidic systems. In order to build these systems, non-invasive actuators such as pumps, filters and mixers are required. We have demonstrated microfluidic sorting based on a 3D interference pattern, formed from multiple coherent laser beams, which has the potential to fulfil all the above criteria. By interfering five laser beams from a fibre laser at 1070 nm, we have formed a 3D optical lattice. When particles flow through the optical lattice their trajectories depend upon the force exerted on the particle by the optical lattice, in combination with the drag force exerted by the fluid flow. Hence, with the strength of a particle's interaction with the lattice determining the total force exerted upon it, its trajectory is determined by its physical properties. These properties include refractive index, size and shape, giving a range of criteria with which to sort an analyte. We have shown separation at 45 degrees of polymer from silica microspheres (by refractive index), the separation of protein microcapsules and the sorting of erythrocytes from lymphocytes. The interference pattern can be tailored to the particles and if a blockage occurs, the laser can simply be switched off, unlike solid-state micro-sorters, so that no jamming occurs. Efficiencies in excess of 95% have been achieved.

Animals↗

Size fractionation and characterisation of fresh water colloids and particles: split-flow thin-cell and electron microscopy analyses.

Split-flow thin-cell (SPLITT) was employed in conventional mode (CSF), to size-fractionate colloids and particles from a selected freshwater. Imaging and quantification by calculations of particle size distributions (PSDs) and shape factors were performed on sample analyzed by conventional high vacuum scanning electron microscopy (SEM) and environmental SEM (ESEM), to investigate the ability of SPLITT to make accurate and nonperturbing separations. SEM and ESEM images of unperturbed and SPLITT-generated fractions were used in order to obtain qualitative and quantitative information about the properties of colloids and particles. Particle size distributions (PSDs) showed that separations were very good, agreeing with theoretical behavior. ESEM PSDs showed that up to 87-88% of the material in the a fraction (expected to be <1 microm) was in fact less than 1 microm and in the b fraction (>1 microm) 87-95% of the material was the expected size. The SEM data indicated a slightly higher contamination of the b fraction with the presence of submicron colloids. Moreover, analysis of conformations indicated significant nonsphericity in unfractionated colloids and particles, but after SPLITT fractionation, shape factors showed that particles were significantly more spherical than before separation.

Chemical Fractionation↗

Fractionation of surface sediment fines based on a coupled sieve-SPLITT (split flow thin cell) method.

In traditional sediment grain-size separation using sieve technique, the bulk of the organic matter passes through the smallest mesh size (generally 38 microm) and is not further fractionated. In this study, a common sieve separation has therefore been coupled with an extra high capacity split flow thin cell fractionation (EHC-SPLITT) instrument to separate the bulk surface sediment not only into size-based sieve fractions (> 100, 63-100, 38-63 and < 38 microm) but particularly to further fractionate hydrodynamically the fine fraction (< 38 microm) using the EHC-SPLITT. Compared to the few previous studies using a smaller high capacity (HC) SPLITT cell, the EHC-SPLITT evaluated in detail here has several advantages (e.g., 23 times higher throughput and allowance for large particle diameters). First, the EHC-SPLITT was calibrated with particle standards. Then, its ability to fractionate fine surface sediments hydrodynamically was demonstrated with material from biogeochemically distinct regimes using two cutoff velocities (1 and 6 m d(-1)). The results from particle standards indicated a good agreement between theory and experiment and a satisfactory mass recovery for the sieve-SPLITT method (80-97%) was observed for sediment samples. The mass distributions revealed that particles < 38 microm were predominant (70-90%), indicating the large need for a technique such as the EHC-SPLITT to further fractionate the fine particles. There were clearly different compositions in the EHC-SPLITT-mediated sub-fractions of the sediment fines as indicated by analyses of organic and inorganic parameters (POC, Si, Fe and Al). The EHC-SPLITT technique has the potential to provide information of great utility to studies of benthic boundary layer transport and off-shelf export and how such processes fractionate geochemical signals.

Calibration↗