Graphene oxide sheets fixed over silica particles (SiGO) and their modification functionalized with C18 and endcapped (SiGO-C18ec) have been reported as sorbents for extraction and analytical columns in LC. In this study, a SiGO column was selected as the extraction column and a SiGO-C18ec as the analytical column to study the applicability and limitations of a column-switching system composed exclusively of columns packed with graphene-based sorbents. Pyriproxyfen and abamectin B1a were selected as the analytes, and orange-flavored carbonated soft drinks as the matrix. The proposed system could be successfully applied to the pyriproxyfen analysis in a concentration range between 0.5 to 25 µg/mL presenting a linearity of R2 = 0.9931 and an intra-day and inter-day accuracy of 82.2-111.4% (RSD < 13.3%) and 95.5-99.8% (RSD < 12.7%), respectively. Furthermore, the matrix composition affected the area observed for the pyriproxyfen: the higher the concentration of orange juice in the soft drink, the higher the pyriproxyfen the signal observed. Additionally, the SiGO extraction column presented a life use of 120 injections for this matrix. In contrast, the proposed system could not apply to the analysis of abamectin B1a, and the SiGO-C18ec analytical column presented significant tailing compared to a similar approach with a C18 analytical column.

In chemical processes, packed columns are frequently employed in various unit operations. However, the flow rates of gas and liquid in these columns are often constrained by the risk of flooding. To ensure the safe and efficient operation of packed columns, it is crucial to detect flooding in real time. Conventional flooding monitoring methods rely heavily on manual visual inspections or indirect information from process variables, which limit the real-time accuracy of results. To address this challenge, we proposed a convolutional neural network (CNN)-based machine vision approach for non-destructive detection of flooding in packed columns. Real-time images of the packed column were captured using a digital camera and analyzed with a CNN model, which was been trained on a dataset of recorded images to identify flooding. The proposed approach was compared with deep belief networks and an integrated approach of principal component analysis and support vector machines. The feasibility and advantages of the proposed method were demonstrated through experiments on a real packed column. The results showed that the proposed method provides a real-time pre-alarm approach for detecting flooding, enabling process engineers to quickly respond to potential flooding events.

Graphene oxide-based LC stationary phases were developed and applied for separating hormones from urine using capillaryLC-MS/MS. Using two analytical approaches - direct injection and column-switching arrangement - it was possible to evaluate the chromatographic parameters and perform tests on the raw biological fluid. Two stationary phases (SPs) were produced, varying the amino silica support particle diameter (Si, 5, and 10 μm). Graphene oxide was covalently bonded to the surface of Si particles, and this material was functionalized by the insertion of octadecylsilica groups, generating the SiGO-C18. Infra-red spectroscopy assays revealed that both steps were successful - supporting GO onto Si and further C18 customization. Scanning electron microscopy showed spherical geometries with minor irregularities and narrow particle size distribution for the produced SPs. The GO-coating rate was higher on the Si particles of 10 μm. As a result, the 10 μm produced column reported better resolution, efficiency, and peak capacity. Therefore, this SiGO-C18 capillary column (100 mm × 0.32 mm i.d., 10 μm dp) was applied successfully in a column-switching method to separate hormones in urine. Linearity (R2 above 0.99), quantification limits (between 1.0 and 5 μg/L), and other figures of merit of the method were determined. It is worth mentioning that the SiGO-C18 capillaryLC column performed adequately, separating the target compounds in less than 6 min. We hope this work could significantly contribute to shedding some light on graphene-based materials as a promising class of stationary phase for miniaturized liquid chromatography.

Often only small amounts of sample are available for spectroscopic analytical determinations. This work investigates the enhancement of signal in columns packed with silica particles. We propose that silica particles cause the light to scatter through the column, effectively increasing optical path length. Packed columns are shown to be effective with fluorescence spectroscopy, but results were inconclusive with absorbance spectroscopy.

The fabrication of effective microchip liquid chromatography (LC) systems tends to be limited by the availability of suitable chromatographic columns. Herein, we developed a glass microchip LC system in which porous single-particle silica was adopted as frits and a freeze-thaw valve was utilized to achieve sample injection without interfering with sampling. The fabrication of single-particle-frit-based packed columns did not require an additional packing channel, thus avoiding dead volumes within the channel interface that can influence chromatographic separation. Further, the length of the packed column could be adjusted using the location of single-particle frits within the column channel. The fabricated frits exhibited high mechanical strength, good permeability, and tolerance for high pressures during chromatographic separation. In particular, the developed microchip LC system was able to withstand high separation pressures of more than 5000 psi. The microchip LC system was applied to the separation of neurotransmitters. Three different monoamine neurotransmitters were completely separated within 5 min with theoretical plate numbers on the order of 100,000 plates m-1. The microchip LC system has a potential for application in a variety of fields including environmental analysis, food safety, drug analysis, and biomedicine.

In this chapter, protocols and details for the immobilization of a model cell onto polyurethane foam carriers are provided in order to facilitate the use of such systems in laboratory or industrial reactors. Polyurethane foam has recently acquired great relevance as a carrier for its good mechanical properties, high porosity, and large adsorption surface. In addition, it has a very low commercial cost. Two different immobilization protocols have been described, differing in the flow regime or the possibilities for the reactor: immobilization in a stirred tank reactor working in a discontinuous regime (by cycles) and immobilization in a packed column working in continuous operation mode. Protocols for carrier sterilization, analytical methodology, and immobilization are described.

The purpose of this work was to compare side by side the performance of packed bed and membrane chromatography adsorption processes for protein purification. The comparison was performed using anion exchange media with the same ligand immobilized on the adsorbing surface, namely the strong Q quaternary ammonium group, R-CH2-N+-(CH3)3, and bovine serum albumin (BSA) as a model protein. In addition, the stationary phase volume was held constant for each geometry (3 mL) and runs were executed using the same mobile phase superficial velocity. As expected, the packed bed column showed higher equilibrium binding of BSA at 66.9 mg/mL versus 43.04 mg/mL for the membrane adsorber. Dynamic binding capacities were also higher in the packed bed; for example, at 97.5 cm/h, a capacity of 62.8 mg/mL was measured for the packed bed versus 20.7 mg/mL for the membrane adsorber. The higher equilibrium and dynamic capacities of the packed bed are likely due to the higher surface area per unit volume of the resin. However, the maximum productivity for the membrane adsorber was 111 mg/(mL h), a value that was 3.3 times higher than the one of the packed column. The bed utilization - defined as the ratio of the dynamic binding capacity at 10% breakthrough to the saturation binding capacity - was also higher for the packed column at long residence times and lower at short residence times confirming the better performance of membrane chromatography at high flow rates.

In this study, an in-tube solid-phase microextraction column packed with mesoporous TiO2 nanoparticles, coupled with MALDI-TOF-MS, was applied to the selective enrichment and detection of phosphopeptides in complex biological samples. The mesoporous TiO2 nanoparticles with high specific surface areas, prepared by a sol-gel and solvothermal method, were injected into the capillary using a slurry packing method with in situ polymerized monolithic segments as frits. Compared with the traditional solid-phase extraction method, the TiO2 -packed column with an effective length of 1 cm exhibited excellent selectivity (α-casein/β-casein/BSA molar ratio of 1:1:100) and sensitivity (10 fmol of a β-casein enzymatic hydrolysis sample) for the enrichment of phosphopeptides. These performance characteristics make this system suitable for the detection of phosphorylated peptides in practical biosamples, such as nonfat milk.

Hydrophilic interaction liquid chromatography, HILIC, is a relatively new HPLC mode. Compared with other HPLC modes, HILIC is a high resolution chromatographic mode with high peak capacity for separations of complex mixtures. Although the separation mechanism is still not completely clear, HILIC has been widely used for analysis of hydrophilic compounds which are difficult for reversed phase chromatography to retain and separate. In this study, we fabricated and investigated nanoHILIC columns in terms of separation efficiency, van Deemter curves and more importantly, we focused on long packed capillary columns, and studied their extreme resolution for protein digests. Using meter long nanoHILIC columns packed with 5 μm particles, we realized a high peak capacity of 130. Based on nanoLC-MS, we compared the resolution and protein identification capabilities of nanoHILIC and nanoRPLC. The results indicate both nanoHILIC and nanoRPLC can provide high resolution for protein sequencing but neither mode is significantly better than the other. Among the 99 digest peptides identified, 17 were uniquely identified by nanoHILIC-MS and 20 were uniquely identified by nanoRPLC-MS and 62 were identified by both methods. Although at this moment in time, nanoRPLC is the most popular microseparation tool in proteomics, the excellent complementarity of nanoHILIC and nanoRPLC suggests their combined use in achieving deep-coverage in MS-based proteomics.

An up-flow fixed column study was conducted to remediate Remazol Brilliant Orange 3R (RBO3R) from contaminated solutions using biochar derived from Ulva lactuca biomass. The influences of column parameters on dye sorption were studied in detail, which include initial RBO3R concentration, bed depth, and flow rate. Optimization experiments indicated that maximum RBO3R column uptake of 0.114 mmol/g was observed at 0.25 mmol/L (initial RBO3R concentration), 0.3 L/hr (flow rate), and 25 cm (U. lactuca bed depth). Modeling of column sorption data was performed using the Yoon-Nelson, modified dose-response and Thomas models. The spent biochar was desorbed and rejuvenated using 0.01 M NaOH. The elutant (0.01 M NaOH) exhibited 99.7% efficiency, and the process was completed in 115 min with high overall concentration factor of 8.4. PRACTITIONER POINTS: This study explores the impact of column parameters on the dye removal potential of U. lactuca-derived biochar. At optimized condition, the biochar bed exhibited highest Remazol Brilliant Orange 3R uptake capacity of 0.114 mmol/g. The regeneration and desorption of U. lactuca-derived biochar bed was possible with NaOH (0.01 M) as elutant.