Disposable microcolumns with welded metal frits

This study reports the preparation of disposable microcolumns with welded metal frits for the first time. First, the bottom of glass‐lined stainless‐steel tubing of 30 cm length, 1.6 mm od, and 0.5 mm id was welded with a stainless‐steel screen frit of 1.6 mm diameter. A micro‐welding machine was used for this. Next, the column was connected to a slurry packer and packed with porous silica particles. Then, the inlet of the column was carefully welded with another frit. The column was tested for separation of a test mix composed of phenol, 2‐nitrophenol, acetophenone, aceanilide, and benzamide. Another column of the same physical dimension was also prepared with frits that were not welded to the column. The chromatographic performances of the two groups of columns (welded frits versus non‐welded frits) were examined. The columns of welded frits showed ca. 18% better separation efficiency (number of theoretical plates) than those of non‐welded frits.     

Organic monolith frits encased in polyether ether ketone tubing with improved durability for liquid chromatography

This study introduces a preparation method for polymer‐encased monolith frits with improved durability for liquid chromatography columns. The inner surface of the polyether ether ketone tubing is pretreated with sulfuric acid in the presence of catalysts (vanadium oxide and sodium sulfate). The tubing was rinsed with water and acetone, flushed with nitrogen, and treated with glycidyl methacrylate. After washing, the monolith reaction mixture composed of lauryl methacrylate, ethylene glycol dimethacrylate, initiator, and porogenic solvent was filled in the tubing and subjected to in situ polymerization. The tubing was cut into thin slices and used as frits for microcolumns. To check their durability, the frit slices were placed in a vial and a heavy impact was applied on the vial by a vortex mixer for various periods. The frits made in the presence of catalysts were found to be more durable than those made without catalysts. Furthermore, when the monolith‐incorporated tubing was used as a chromatography column, the column prepared in the presence of catalysts resulted in a better separation efficiency. The separation performance of the columns installed with the polyether ether ketone encased monolith frits was comparable to that of the columns installed with the commercial stainless‐steel screen frits.     

Polyether ether ketone encased monolith frits made of polyether ether ketone tubing with a 0.25 mm opening resulting in an improved separation performance in liquid chromatography

Tiny polyether ether ketone encased monolith frits have been prepared by modified catalytic sulfonation of the inner surface of polyether ether tubing (1.6 mm od, 0.25 mm id) followed by modified formation of organic monolith and cutting of the tubing into slices. The frit was placed below the central hole of the column outlet union and supported by a combination of a silica capillary (0.365 mm od, 0.05 mm id) and a polyether ether ketone sleeve (1.6 mm od, 0.38 mm id) tightened with a nut and a ferrule when the column was packed to prevent sinking of the frit element into the union hole (0.25 mm opening) otherwise. The column packed this way with the frits investigated in this study has shown better separation performance owing to the reduced frit volume in comparison to the column packed with a commercial stainless‐steel screen frit. This study establishes the strategy of disposable microcolumns in which cheap disposable frits are used whenever the column is re‐packed to yield columns of even better chromatographic performance than the columns with commercial frits.     

Coating properties of a novel water stationary phase in capillary supercritical fluid chromatography

The coating properties of a novel water stationary phase used in capillary supercritical fluid chromatography were investigated. The findings confirm that increasing the length or internal diameter of the type 316 stainless‐steel column used provides a linear increase in the volume of stationary phase present. Under normal operating conditions, results indicate that about 4.9 ± 0.5 μL/m of water phase is deposited uniformly inside of a typical 250 μm internal diameter 316 stainless‐steel column, which translates to an area coverage of about 6.3 ± 0.5 nL/mm² regardless of dimension. Efforts to increase the stationary phase volume present showed that etching the stainless‐steel capillary wall using hydrofluoric acid was very effective for this. For instance, after five etching cycles, this volume doubled inside of both the type 304 and the type 316 stainless‐steel columns examined. This in turn doubled analyte retention, while maintaining good peak shape and column efficiency. Overall, 316 stainless‐steel columns were more resistant to etching than 304 stainless‐steel columns. Results indicate that this approach could be useful to employ as a means of controlling the volume of water stationary phase that can be established inside of the stainless‐steel columns used with this supercritical fluid chromatography technique.     

Silmization of stainless-steel frits for use in trace metal analysis by high performance liquid chromatography

A simple method has been developed to minimize the interferences related to the metal parts of an analytical system used for trace metal analysis by HPLC. This is accomplished by blocking the exposed metal surfaces with an organosilane. The effect of the silanized stainless steel frits has been demonstrated by injecting diethyldithiocarbamate into columns with and without high metal surface area components. The coating of the silane is quite stable for routine use.     

Visualization of solute migration in chromatographic columns influence of the frit porosity

The dual-perspective, on-column detection method previously described was used to observe the effects of the inlet frit on the profiles of chromatographic bands. Visualization of bands of iodine was achieved by injecting its dilute solutions in carbon tetrachloride into a glass column packed with a C18-bonded silica and eluted with carbon tetrachloride, which has the same refractive index as the packing material. The bands were photographed on-column with two standard 35-mm SLR cameras oriented at right angle. The photographs were scanned and the digitized images of the sample bands analyzed with proper software. A number of columns, as similar as possible, were fitted with different 2- and 10-microm porosity stainless steel frits. Subsequent analysis of the digitized band images revealed irregularities in the band shape resulting from frit contributions to band dispersion. The 2-microm frits produced more dramatic effects overall than the coarser frits. Local axial dispersion coefficient values, expressed as local reduced plate height, were calculated. The results demonstrate the possibly damaging effects of the frit on the band shape.     

A Simple Method to Prepare On‐Capillary Frits for Micro‐Column Separation

Sintering stainless steel powders was initially used to prepare the inlet frit in fused silica capillaries. The use of such inlet frits and outlet frits sintered by the stationary phase itself in the capillary to retain C₁₈ particles was demonstrated to withstand the long exposure, up to a high pressure of 60 MPa, for packing and the prepare column was stable and robust enough to do the continuous chromatographic operations. Characterization of the inlet and outlet frits by scanning electron micrography showed the fused metal particles formed a porous network in the capillary inlet and the homogenous separation beddings were obtained by slurry packing.     

Adsorption on stainless steel surfaces of biosurfactants produced by gram-negative and gram-positive bacteria: Consequence on the bioadhesive behavior of Listeria monocytogenes

The ability of adsorbed biosurfactants (Pf and Lb) obtained from gram-negative bacterium (Pseudomonas fluorescens) or gram-positive bacterium (Lactobacillus helveticus) to inhibit adhesion of four listerial strains to stainless steel was investigated. These metallic surfaces were characterized using the following complementary analytical techniques: contact-angle measurements (CAM), atomic force microscopy (AFM), polarization modulation-infrared reflection-adsorption spectroscopy (PM-IRRAS) and X-ray photoelectron spectroscopy (XPS). Contact-angles with polar liquids (water and formamide) indicated that the stainless steel surface covered with adsorbed biosurfactant was more hydrophilic and electron-donating than bare stainless steel. The surface characterization by XPS and PM-IRRAS revealed that conditioning the stainless steel changes the substrate in two ways, by modifying the surface alloy composition and by leaving an thin adsorbed organic layer. AFM observations enabled to say that the layer covered entirely the surface and was probably thicker (with patches) in the case of Pf-conditioned surfaces compared to the Lb-conditioned ones, which seemed to be less homogeneous. Though the added layer was thin, significant chemical changes were observed that can account for drastic modifications in the surface adhesive properties. As a matter of fact, adhesion tests showed that both used biosurfactants were effective by decreasing strongly the level of contamination of stainless steel surfaces by the four strains of Listeria monocytogenes. The more important decrease concerned the CIP104794 and CIP103573 strains (>99.7%) on surface conditioned by L. helveticus biosurfactant. A less reduced phenomenon (75.2%) for the CIP103574 strain on stainless steel with absorbed biosurfactant from P. fluorescens was observed. Whatever the strain of L. monocytogenes and the biosurfactant used, this antiadhesive biologic coating reduced both total adhering flora and viable and cultivable adherent bacteria on stainless steel surfaces. This study confirms that biosurfactants constitute an effective strategy to prevent microbial colonization of metallic surfaces by pathogenic bacteria like the food-borne pathogen L. monocytogenes.     

筛板对台锥形制备液相色谱柱流型和柱效的影响

制备液相色谱柱柱头筛板的选择直接影响到样品在柱头的分配情况及色谱柱柱效的高低。 利用柱后可见-紫外检测,比较了采用多孔聚四氟乙烯材料和多孔烧结不锈钢材料作为台锥形制备液相色谱柱柱头筛板材料时的柱效;用柱上可视化方法研究了柱头筛板的直径对流型的影响。结果表明,多孔聚四氟乙烯的变形使得采用该材料做柱间筛板的色谱柱柱效比使用多孔不锈钢筛板的色谱柱柱效低约40%;样品流经直径小于柱入口内径的筛板导致样品在柱头分配不均,而流经直径与柱入口内径相同的筛板时因受到的阻力相同,可以实现柱塞状进样。还观察到不同直径的筛板     

Sample handling and contamination encountered when coupling offline normal phase high performance liquid chromatography fraction collection of petroleum samples to Fourier transform ion cyclotron resonance mass spectrometry

Normal phase high performance liquid chromatography (HPLC) is used to separate a gas oil petroleum sample, and the fractions are collected offline and analyzed on a high resolution Fourier Transform Ion Cyclotron Resonance Mass Spectrometer (FT-ICR MS). The separation prior to MS analysis dilutes the sample significantly; therefore the fractions need to be prepared properly to achieve the best signal possible. The methods used to prepare the HPLC fractions for MS analysis are described, with emphasis placed on increasing the concentration of analyte species. The dilution effect also means that contamination in the MS spectra needs to be minimized. The contamination from molecular sieves, plastics, soap, etc. and interferences encountered during the offline fraction collection process are described and eliminated. A previously unreported MS contamination of iron formate clusters with a 0.8 mass defect in positive mode electrospray is also described. This interference resulted from the stainless steel tubing in the HPLC system. Contamination resulting from what has tentatively been assigned as palmitoylglycerol and stearoylglycerol was also observed; these compounds have not previously been reported as contaminant peaks.     

Zinc deposition during ESI-MS analysis of peptide-zinc complexes

Electrospray ionization (ESI) mass spectrometry (MS) has proven to be an extremely powerful technique for studying the stoichiometry and binding strength of peptide-metal complexes. We have found a significant new problem in the ESI-MS of zinc-peptide systems involving the deposition of zinc in the ESI emitter. This deposition of zinc during the experiment removes a significant amount of zinc ions from the solution, impacting the resulting mass spectral intensities used to quantify the amount of the zinc-bound species. Analysis of infused zinc-peptide samples with atomic absorption spectrometry and with a custom-built nanoflow ESI source confirms the alteration of the analyte solutions with positive or negative or no potential applied to the emitter. Ultimately, the location of the zinc deposition was determined to be the stainless steel emitter. The use of a custom-built nanoESI interface using glass emitters was found to mitigate the zinc deposition problem. The phenomenon of metal deposition warrants further investigation as it may not be limited to just zinc and may represent a significant obstacle in the ESI-MS analysis of all protein-metal systems.     

Electronic tongue based on an array of metallic potentiometric sensors

An electronic tongue system based on the array of six metallic potentiometric sensors (metallic wires) was developed and utilized for discrimination of foodstuffs: several types of vinegar and fruit juices. Copper, tin, iron, aluminum, brass and stainless steel wires were included in the array and supplemented by pH glass electrode. The response of potentiometric metallic sensors towards various organic acids has been studied and possible sensitivity mechanisms were discussed. Overall potential changes of metallic sensors were exanimate as complex mixed signals influenced by several components presenting in analyte employing chemometric approach. The multisensor array of such a type can be useful for several applications since of simplicity in handling, low cost of sensors and easy measure procedure.     

Monitoring metal ion contamination onset in hydrofluoric acid using silicon-diamond and dual silicon sensing electrode assembly.

Potentiometric detection of trace levels of metallic contamination onset in hydrofluoric acid using a silicon-based sensor in conjunction with two non-contaminating reference electrode systems is presented in this paper. In the first case, conductive diamond was used as a non-contaminating reference electrode. Cyclic voltammetry and open-circuit potential experiments demonstrated the feasibility of using a conductive diamond film electrode as a quasi-reference electrode in the HF solution. In the second case, a dual silicon electrode system was used with one of the silicon-based electrodes protected with an anion permeable membrane behaving as the quasi-reference electrode. The dual silicon sensing electrode system possessed an additional operational advantage of being unaffected by the solution acidity. Though both sensing configurations were able to detect the metal ion contamination onsets at the parts-per-trillion to parts-per-billion levels, the dual silicon electrode design showed a greater compatibility for the on-line detection of metallic impurities in HF etching baths commonly used in semiconductor processing.     

Metal-based impurities in graphenes: application for electroanalysis

We show here that metallic impurities presented in graphenes prepared from graphite can be usefully employed for electroanalysis. We demonstrate that cumene hydroperoxide electrochemical reduction on graphene containing iron-based impurities provides significantly larger voltammetric currents than the same experiment using iron oxide nanoparticles. This opens doors for turning metallic impurities into potentially useful components of graphene based electrochemical systems.     

Origin and correction of the deviations in retention times at increasing flow rate with Chromolith columns

Chromoliths can be used at flow rates beyond those feasible for conventional microparticulate packed columns. Ideally, the plots of the retention time versus the inverse of delivered flow rate should exhibit y-intercept of zero. However, significant positive deviations correlating with the solute polarity were observed for several compounds chromatographed with a Chromolith column, owing to the increased system pressure. Consequently, the dead time marker exhibits a smaller deviation, making the retention factors depend on the flow rate. Chromoliths are made of a silica-based monolith encapsulated within a PEEK tube, and should suffer larger stress with pressure than stainless steel columns, tending to inflate them and increase their volume. This decreases the linear velocity inside the column, and increases the retention at relatively low pressure (<200 bar). In contrast, frictional heating, which is an issue for microparticulate columns, seems to be less significant for the highly permeable Chromoliths. The usefulness of the retention time versus the inverse of the delivered flow rate plots to measure the deviations, whatever their origin, is shown. This allows the correction of the retention times to the ideal behaviour, where the retention factors are independent of the flow rate.     

A preliminary comparative assessment by activation analysis of some of the materials and containers used in pharmaceutical manufacture

Neutron activation analysis has been used in a preliminary, comparative assessment of some of the materials and containers employed in pharmaceutical manufacture. Microgram and submicrogram amounts of manganese, copper, and sodium have been determined in water distilled and redistilled from stainless steel and pyrex glass stills. The leaching of these elements by distilled and doubly distilled water from stainless steel, pyrex glass, and polyethylene storage containers has also been studied. The Mn, Cu, and Na contents of polyethylene bottles were also determined. The same techniques were used to determined these impurities as well as As, Au, and Sb in adrenaline and vitamin C.     

Silicate entrapped columns--new columns designed for capillary electrochromatography

Designed especially for capillary electrochromatography (CEC), silicate-entrapped columns are made by trapping particles of chromatographic packing material in a network of silica. Once entrapped, the capillary no longer requires frits. This renders a more homogeneous and stable packed bed. Accidental breakage of the fragile frits is not an issue with these robust columns. Columns packed with reverse-phase material subjected to silicate entrapment demonstrated faster separations of retained analytes and increased efficiencies compared with nonentrapped columns. The method was also used to prepare chiral CEC columns by entrapping a molecular imprinted polymeric (MIP) packing having minimal surface charge density, thus being unable alone to support sufficient electroosmotic flow for CEC.     

The Significance of Metallophilic and Silanophilic Interactions is Reversed Phase Hplc

Abstract

The poor peak shape encountered for amines in reversed phase high performance liquid chromatography (RPLC) is widely recognized. In order to improve chromatographic efficiency, the mechanism of the broadening must be understood. In addition to the silanol groups on the support surface, raetal sites have been implicated as possible adsorption sites for amines.

We have investigated the relative role of metallophilic and silanophilic interactions in the non-ideal behavior of amines and proteins. Stainless steel frits appear to have a deleterious effect on column efficiency via both mechanical and chemical interactions. The use of stainless steel meshes or screens is recommended. The residual metal sites on the support material have been found to play an insignificant role in solute retention or peak broadening. Minimization of silanophilic interactions is therefore the key to increasing efficiency in the separation of basic amines.     

Molecularly imprinted polymer grafted to porous polyethylene frits: a new selective solid-phase extraction format

In this paper, a novel format for selective solid-phase extraction based on a molecularly imprinted polymer (MIP) is described. A small amount of MIP has been synthesized within the pores of commercial polyethylene (PE) frits and attached to its surface using benzophenone (BP), a photo-initiator capable to start the polymerisation from the surface of the support material. Key properties affecting the obtainment of a proper polymeric layer, such as polymerisation time and kind of cross-linker were optimised. The developed imprinted material has been applied as a selective sorbent for cleaning extracts of thiabendazole (TBZ), as model compound, from citrus samples. The use of different solvents for loading the analyte in the imprinted frits was investigated, as well as the binding capacity of the imprinted polymer. Imprinted frits showed good selectivity when loads were performed using toluene and a linear relationship was obtained for the target analyte up to 1000 ng of loaded analyte. Prepared composite material was applied to the SPE of TBZ in real samples extracts, showing an impressive clean-up ability. Calibrations showed good linearity in the concentration range of 0.05-5.00 μg g(-1), referred to the original solid sample, and the regression coefficients obtained were greater than 0.996. The calculated detection limit was 0.016 μg g(-1), low enough to satisfactory analysis of TBZ in real samples. RSDs at different spiking levels ranged below 15% in all the cases and imprinted frits were reusable without loss in their performance.     

Magnetically immobilized frits for the preparation of packed columns used in capillary electrochromatography

Fabrication of porous frits to retain stationary phases is a critical issue in column preparation for capillary electrochromatography (CEC). In this work, porous frits were prepared by applying an external magnetic field to magnetically responsive particles placed inside a fused-silica capillary. Three batches of uniform magnetite spheres with particle diameters of 0.3, 0.4, and 0.6 μm and saturation magnetization values of 73.03, 74.41, and 77.83 emu/g, respectively, were used as frit particles and octadecyl- and phenyl-bonded silica gels were packed successfully into frit-containing capillaries. The performance of the resulting magnetically immobilized frits and packed columns was evaluated. The electroosmotic mobilities in capillaries containing outlet frit only were found to be reduced by 2–4% whereas the plate heights of an unretained marker increased by 30–50% as compared to those in open capillaries. These variations are believed to be associated with the inhomogeneities of the packed structure of the frits. The magnetically immobilized frits showed adequate mechanical strength to withstand the flow drag force, allowing separation in capillaries packed with 5-μm stationary phases up to 10–15 cm, thus rendering column efficiency and reproducibility comparable with those obtained with sintered frits. Taken together, retaining frits made of uniform magnetite particles serves as a viable alternative to sintered frits for column preparation, which offers several distinct advantages such as ease of preparation, improved durability as compared to sintered frits where the removal of the polyimide coating makes the packed column susceptible to breakage, and use of large-bore capillaries for semipreparative separations.