Column Technology Research Articles

High throughput and cost-effective purification of small molecules using superficially porous stationary phase columns: An environment friendly approach

Over the past two decades, higher efficiencies have become the goal of most research organizations, and more so for high throughput synthesis. Consequently, there is an urgent need for an advanced purification approach to accommodate the large number of small molecules in a shorter turnaround time. To address this, an efficient and high throughput purification method was developed leveraging the superficially porous particle (SPP) column technology, which was optimized with respect to detector acquisition rate, flow rate, and gradient time. The experiments were performed using a sample mixture of six commercially available small molecules including Nicotinamide (NIC), 4-Amino benzophenone (ABP), Praziquantel (PRA), butyl‑4-hydroxybenzoate (BHB), Warfarin sodium (WAR), and Ibuprofen (IBU). The study outcomes were promising and exhibited a good separation of all six analytes with ∼83 % less turnaround time and ∼65 % less solvent consumption than the regular methods. Furthermore, it showed significant improvement in peak symmetry, capacity factor, and resolution for all six analytes and afforded superior chromatographic performance than the regular method with fully porous particle (FPP) columns. The study concludes that SPP columns provide cost-effective, eco-friendly, and high throughput purification of small molecules.

Targeted microscopic rare cell retrieval yields sufficient deoxyribonucleic acid for molecular testing in fixed tissue with low tumor fraction

Abstract Introduction/Objective DNA extracted from formalin-fixed paraffin-embedded tissue (FFPE) blocks is often utilized for molecular oncologic testing. Cases with low tumor fraction often fail to obtain adequate tumor for molecular testing. We hypothesized that targeted retrieval of visualized tumor cells from FFPE sections, including those with low tumor fraction, could yield sufficient DNA for molecular testing. Methods/Case Report Unstained sections of various thicknesses were cut from low tumor fraction blocks and visualized by light microscopy and the RareCyte CyteFinder II (Seattle, WA). Microscopic cell retrieval was performed on the RareCyte using 40 µm diameter needles from identified regions of interest. Samples were deparaffinized using Zymo Research (Tustin, CA) solution, and digested and purified using Zymo-Spin Column technology. Sample DNA was quantified with NanoDrop Spectrophotometer (Chino, CA). Results (if a Case Study enter NA) Regions of interest were morphologically identifiable on 4-10 µm thick sections and harder to visualize on 12 µm thick sections. Targeted microscopic cell retrieval was performed with 40 µm diameter needles. Retrieval was performed from 26 to 34 regions of interest per sample and pooled for molecular testing. Samples (n=4) yielded a mean of 81.0 ng of DNA ± 18.4 ng (1.8 ng/µL ± 0.4 ng/µL) and a median of 87.75 ng (1.95 ng/µL). Conclusion Targeted microscopic cell retrieval from low tumor fraction FFPE slides using the RareCyte CyteFinder II can provide sufficient DNA material for molecular testing. Application of this technology has the possibility of recovering sufficient tumor DNA for molecular testing from specimens that may be deemed unsuitable for molecular testing according to current standards. We aim to focus on increasing sample size and completing comprehensive molecular analysis as we continue to expand this work.

Highly Efficient LC‐MS/MS Analysis of Organophosphorous Pesticides Utilizing ARC‐18 Column Selectivity with Inert Column Technology

Highly Efficient LC‐MS/MS Analysis of Organophosphorous Pesticides Utilizing ARC‐18 Column Selectivity with Inert Column Technology

Cyclic response of floating geosynthetic-encased steel slag columns in soft clay

Geosynthetic-encased stone column (GESC) technology for strengthening soft clay offers significant advantages in terms of cost-effectiveness, environmental sustainability, and engineering applicability. It is widely applied in treating soft foundations for railways, bridges, and embankments. This study evaluates the cyclic response of the geosynthetic-encased steel slag column (GESSC) composite foundation employing three-dimensional nonlinear finite element analysis. A numerical study is conducted to assess the cyclic response of floating GESSC considering the influence of key design variables, including cyclic load amplitude, loading frequency, geosynthetic encasement stiffness, and length-to-diameter ratio. Results show that both cyclic load amplitude and frequency affect the cumulative settlement and excess pore pressure within the GESSC foundation. Within specified limits, increasing the encasement stiffness and column length can significantly improve the GESSC load-bearing characteristics. The parametric study suggests an optimal geosynthetic encasement stiffness for the field prototype columns within the range of 4480–5760 kN/m and a critical steel slag column length of 10 times the column diameter.

Technical Aspects of Natural Gas Pyrolysis in Liquid Metal Bubble Column Reactors

The pyrolysis of low alkanes (in the following short “pyrolysis”) has already been investigated during the 1960s. However, none of the reactor systems used at the time are capable of continuous operation. Therefore, the Karlsruhe Institute of Technology has intensified the development of the promising liquid metal bubble column technology in recent years, which is capable of continuous operation. Various key aspects have been addressed, such as scale‐up and the pyrolysis of high‐caloric natural gas. Herein, further developments for a pilot scale system have been investigated, which concern increased throughput and long‐term operation capabilities. Careful evaluation of the impact of according measures has been done, which shows that the achieved scale‐up has only negligible effects on the pyrolysis outcome. The effects of the scale‐up on residence times are negligible. The bubble formation behavior depends on the throughput and the characteristics of the orifice. Wall effects are marginal. Fundamental minimization of weeping could not be confirmed. Reactor pre‐chambers in combination with tin collection chambers are recommended for further scale‐up. An increase in the volume flow should be examined. In terms of long‐term operation , head as well as feed pressure control is recommended.

Simultaneous analysis of pesticides and mycotoxins in primary processed foods: The case of bee pollen

Primary Processed Foods are a class of food items that are ready for consumption after minimal processing in the supply chain. These products are ubiquitous in our daily diet, but so far a limited number of studies dealt with the optimization of quality control methods to check their content of contaminants. Among primary processed foods, bee pollen is a nutritionally acclaimed food supplement, whose contamination with pesticides and mycotoxins has been largely proven. For this reason, the present study aimed at optimizing for the first time a comprehensive LC-MS/MS method capable of analyzing 282 pesticides and 8 mycotoxins in bee pollen. To obtain a suitable method, two extraction procedures (QuEChERS and Accelerated Solvent Extraction), as well as different chromatographic gradients and columns, were tested. The optimized methodology, comprehending an extraction based on semi-automated QuEChERS, and an analytical method including inert LC column technology, was validated and applied to a sample set of 34 bee pollens. The analyzed samples collectively showed the presence of 41 pesticides and 1 mycotoxin.

Improving the intestinal lipidome coverage in a gnotobiotic mouse model using UHPLC-MS-based approach through optimization of mobile phase modifiers and column selection

Lipidomics is focusing on the screening of lipid species in complex mixtures using mass spectrometry-based approaches. In this work, we aim to enhance the intestinal lipidome coverage within the Oligo-Mouse-Microbiota (OMM12) colonized mouse model by testing eight mobile phase conditions on five reversed-phase columns. Our selected mobile phase modifiers included two ammonium salts, two concentrations, and the addition of respective acids at 0.1 %. We compared two columns with hybrid surface technology, two with ethylene bridged hybrid technology and one with core–shell particles. Best performance was attained for standards and intestinal lipidome, using either ammonium formate or acetate in ESI(+) or ammonium acetate in ESI(−) for all column technologies. Notably, a concentration of 5 mM ammonium salt showed optimal results for both modes, while the addition of acids had a negligible effect on lipid ionization efficiency. The HST BEH C18 column improved peak width and tailing factor parameters compared to other technologies. We achieved the highest lipid count in colon and ileum content, including ceramides, phosphatidylethanolamines and phosphatidylcholines, when using 5 mM ammonium acetate in ESI(−). Conversely, in ESI(+) 5 mM ammonium formate demonstrated superior coverage for diacylglycerols and triacylglycerols.

Design comparison and effects of a chamber structure on wave dissipation

ABSTRACT Among all the ocean energy sources that have been widely studied, chamber structures such as oscillating water column (OWC) technologies are effective and low-cost methods to develop hundreds of wave power generation devices as the disadvantage is its lack of efficiency. Even if the OWC system does not perform any power generation operations, it is a wave-absorbing structure. A large part of the wave energy may be blocked by the structure’s acting chamber or could be dissipated during the interaction of the waves in the air chamber, which affects the actual power generation benefit evaluation. Nevertheless, a better wave attenuation structure may present a relatively poor design of the OWC generator chamber. A series of experiments were conducted with regular and irregular wave conditions to study the effects of energy dissipation caused by a chamber structure under varied barrier and orifice designs.

Analytical Quality by Design as applied to the development of a SEC-HPLC platform procedure for the determination of monoclonal antibody purity without mobile phase additives

This work presents the application of AQbD principles to the development of a size exclusion chromatography (SEC) HPLC procedure for the determination of monoclonal antibody (mAb) product purity using state-of-the-art column technology available via the Waters™ XBridge Premier Protein SEC column. Analytical Quality by Design (AQbD) emphasizes a systematic, risk-based lifecycle approach to analytical procedure development based on sound statistical methodologies. It has recently become increasingly recommended by regulatory agencies as a response to the need for greater efficiency, improved reliability, and increased robustness among modern analytical procedures in the pharmaceutical industry. Use of an Analytical Target Profile (ATP) and formal risk assessments informed the application of Design of Experiments (DoE) to optimize this analytical procedure, as well as assess its robustness and ruggedness. Importantly, our ruggedness results demonstrated the transferability of this procedure between two laboratories within the Catalent Biologics Global Network. Application of this analytical procedure as a platform approach for evaluating mAb purity is expected to support expedited, first-in-human timelines of mAb molecules by enabling great quantitative performance with simple mobile phase buffer compositions. Taken together, this case study demonstrates the utility of adopting AQbD principles in analytical procedure development.

Low Phase Ratio Stationary Phase Column Technology for the Characterization of Highly Volatile and Reactive Compounds by Gas-Liquid Chromatography

Recent advances in coating techniques and improved polymers employed as gas chromatographic stationary phases have resulted in commercializing low phase ratio capillary column technology with respectable chromatographic efficiency and inertness. Highly volatile compounds that are critical in challenging industrial applications such as alkanes, alkenes, arsine, phosphine, oxygenated, and sulfur compounds have been successfully analyzed with respectable chromatographic performance and resolution using a low phase ratio column such as a 60 m × 0.32 mm id × 8 µm polydimethylsiloxane (PDMS) with a β value of 10. An R value of 5.6 was achieved for the separation of hydrogen sulfide and carbonyl sulfide without the use of cryogenic chromatography. In addition to improved retention for highly volatile organic compounds, a low phase ratio column can be employed effectively to enhance chromatographic inertness for reactive analytes. The augmentation of a 3D-printed two stage microreactor for use with flame ionization detection enabled carbon compound independent response, thereby lower cost-of-ownership and improved lab safety.

Acetic Acid Enables Molecular Enumeration of Mycobacterium tuberculosis from Sputum and Eliminates the Need for a Biosafety Level 3 Laboratory.

Improved monitoring of Mycobacterium tuberculosis response to treatment is urgently required. We previously developed the molecular bacterial load assay (MBLA), but it is challenging to integrate into the clinical diagnostic laboratory due to a labor-intensive protocol required at biosafety level 3 (BSL-3). A modified assay was needed. The rapid enumeration and diagnostic for tuberculosis (READ-TB) assay was developed. Acetic acid was tested and compared to 4 M guanidine thiocyanate to be simultaneously bactericidal and preserve mycobacterial RNA. The extraction was based on silica column technology and incorporated low-cost reagents: 3 M sodium acetate and ethanol for the RNA extraction to replace phenol-chloroform. READ-TB was fully validated and compared directly to the MBLA using sputa collected from individuals with tuberculosis. Acetic acid was bactericidal to M. tuberculosis with no significant loss in 16S rRNA or an unprotected mRNA fragment when sputum was stored in acetic acid at 25°C for 2 weeks or -20°C for 1 year. This novel use of acetic acid allows processing of sputum for READ-TB at biosafety level 2 (BSL-2) on sample receipt. READ-TB is semiautomated and rapid. READ-TB correlated with the MBLA when 85 human sputum samples were directly compared (R2 = 0.74). READ-TB is an improved version of the MBLA and is available to be adopted by clinical microbiology laboratories as a tool for tuberculosis treatment monitoring. READ-TB will have a particular impact in low- and middle-income countries (LMICs) for laboratories with no BSL-3 laboratory and for clinical trials testing new combinations of anti-tuberculosis drugs.

Reliable Solder Columns to Replace Solder Balls in Large 2.5D Heterogeneous Packages

Solder balls are well suited for interconnecting normal sized ball grid array (BGA) components to printed circuit boards (PCB). However, solder balls are prone to cracking (failure) caused by excessive stresses inherently found in advanced packaging such as large-sized heterogeneous 2.5D packages where multiple dies are placed side-by-side. The growth trend is to make larger and larger BGA processors exceeding 60x60mm in size for AI and data centers. A plethora of new processor substrate and interposer designs are coming on line such as Chip-on-Wafer-on-Substrates (CoWoS). For that reason, more instances of solder ball delamination and cracking on large-sized BGA packages are probable. Solder columns were produced more than 50 years ago to reduce cracking in large ceramic packages, especially for use in harsh environments found in aerospace and defense applications. However, such legacy solder column technology is limited to Tin-Lead (SnPb) alloys. The challenge is to make Lead Free solder columns that satisfy demand for RoHS compliance, while still providing adequate stress relief (compliancy) for extending the operational life of large sized heterogeneous 2.5D processors for mission critical applications. This paper describes our ongoing work to innovate non-collapsible, compliant Lead-Free Braided Copper Solder Columns to reduce strain caused by the Coefficient of Thermal Expansion (CTE) mismatch between large BGA packages and FR4 PCBs. A braid, consisting of 16 fine copper alloy wires, serves as an exoskeleton sleeve around a solder column to provide mechanical support as well as compliancy. This novel solder column technology supports market trends to scale up the size of chip packaging while maintaining reliability. Our focus was to quickly characterize Lead-Free solder column core materials and braiding alloys by employing “Fail-Fast” destructive accelerated mechanical bend testing in our lab. Since there is an ongoing interest in Tin-Lead alloys, this paper also discusses data on Tin-Lead alloys for comparative purposes.

Chromatographic separation by RPLC-ESI-MS of all hydroxyproline isomers for the characterization of collagens from different sources

During collagen biosynthesis, proline is post-translationally converted to hydroxyproline by specific enzymes. This amino acid, unique to collagen, plays a crucial role in stabilizing the collagen triple helix structure and could serve as an important biomarker for collagen content and quality analysis. Hydroxyproline has four isomers, depending on whether proline is hydroxylated at position 4 or 3 and on whether the cis- or trans- conformation is formed. Moreover, as extensive hydrolysis of collagen is required for its amino acid analysis, epimerization may also occur, although to a lesser extent, giving a total of eight possible isomers.The aim of the present study was to develop a reversed-phase high-performance liquid chromatography-UV-mass spectrometry (RPLC-UV-MS) method for the separation and quantification of all eight hydroxyproline isomers. After the chiral derivatization of the hydroxyproline isomers with Nα-(2,4-dinitro-5-fluorophenyl)-L-valinamide (L-FDVA), to enable their UV detection, the derivatized diastereoisomers were separated by testing different C18 column technologies and morphologies and optimizing operative conditions such as the mobile phase composition (solvent, additives), elution mode, flow rate and temperature. Baseline resolution of all eight isomers was achieved on a HALO® ES-C18 reversed-phase column (150×1.5 mm, 2.7 μm, 160 Å) using isocratic elution and MS-compatible mobile phase.The optimized method was validated for the quantification of hydroxyproline isomers and then applied to different collagen hydrolysates to gain insight and a deeper understanding of hydroxyproline abundances in different species (human, chicken) and sources (native, recombinant).

Progress of ligand-modified agarose microspheres for protein isolation and purification.

Proteins arethe material basis of life and the primary carriers of life activities, containing various impurities that must be removedbefore use. To keep pace with the increasing complexity of protein samples, it is essential to constantly work on developing new purification technologies for downstream processes. While traditional downstream purification methods rely heavily on protein A affinity chromatography, there is still a lot of interest in finding safer and more cost-effective alternatives to protein A. Many non-affinity ligands and technologies have also been developed in biological purification recently. Here, the current status of biotechnology and the progress of protein separation technology from 2018 to 2023 are reviewed from the aspects of new preparation methods and new composite materials of commonly used separation media. The research status of new ligands with different mechanisms of action was reviewed, including the expanded application of affinity ligands, the development prospect of biotechnology such as polymer grafting, continuous column technology, and its new applications.

Analytical and experimental study on shaking effects for improved stone column foundations

Stone column technology is a method used to reduce liquefaction in saturated sediments. By using materials with greater permeability than the surrounding soil, stone columns can help drain excess pore water pressure and mitigate liquefaction potential during seismic loading. In this study, analytical solutions are proposed to address the liquefaction problem in saturated soils during earthquakes, taking into account the influence of stone columns and the surrounding soil. These solutions consider the excess pore water pressure at the boundary of the zones and analyze the vertical and horizontal drainages. The study also includes centrifuge tests on sandy soils improved with stone columns, calculating the total seepage flow and converting it into settlement based on the relationship between drainage flow rate and volumetric changes in the soil. The calculated settlement aligns well with the measured value in the centrifuge test, confirming the accuracy of the analytical formula for calculating settlement in stone column foundations. The study also evaluates the combined effects of radial and vertical drainages in stone column foundations using the derived analytical solutions.

A brief review on recent advances in reverse phase HPLC

Modern chromatography is mostly used for analytical purposes. Modern pharmaceutical and biological analysis uses HPLC. It separates molecules from heterogeneous liquids and characterizes them. Mass spectrometry identifies peptides, proteins, and other compounds after high-performance liquid chromatography. HPLC's flexibility and dependability (pressure-driven liquid support) make it popular. However, fast separation often creates high operating pressure, which stresses HPLC equipment. Thus, core-shell silica microspheres and zirconia packing have been produced recently. Fused-core or superficially porous microspheres have solid cores with porous shells. Despite its low pressure, this device can separate small, large, and intricate molecules. It is effective and fast. UPLC offers greater resolution, sensitivity, and separation times than HPLC; however, it may be less reproducible. Rapid Resolution Liquid Chromatography (RRLC) is more precise and sensitive than HPLC, improving sample analysis quality and quantity. This review will highlight column technology basics and RP-HPLC advances and focuses on the concept of HPLC.

Energy-saving design and optimization of pressure-swing-assisted ternary heterogenous azeotropic distillations

A huge amount of energy is always consumed to separate the ternary azeotropic mixtures by distillations. The heterogeneous azeotropic distillation and the pressure-swing distillation are two kinds of effective technologies to separate heterogeneous azeotropes without entrainer addition. To give better play to the synergistic energy-saving effect of these two processes, a novel pressure-swing assisted ternary heterogeneous azeotropic distillation process is proposed firstly. In this process, the ternary heterogeneous azeotrope is decanted into two liquid phases before refluxed into the azeotropic distillation column to avoid the aqueous phase remixing, and three columns’ pressures are modified to decrease the flowrates of the recycle streams. Then the dividing wall column and heat integration technologies are introduced to further reduce its energy consumption, and the pressure-swing assisted ternary heterogeneous azeotropic dividing wall column and its heat integration structure are achieved. Genetic Algorithm procedure is employed to optimize the proposed processes. The design results show that the proposed processes have higher energy efficiencies and lower CO2 emissions, compared with the published ternary heterogeneous azeotropic distillation process.

Sea Wave Energy Simulation of Oscillating Water Column (OWC) Technology, Binongko Island, Wakatobi, Southeast Sulawesi

Indonesia is a country that has the largest sea area of several countries spread across the world. This is an advantage for Indonesia in utilizing the energy potential of the seas and oceans. One of which is the potential of Oscillating Water Column (OWC) Technology, to utilize the energy potential of the seas and oceans not only requires sea potential but also must have location parameters with suitable seabed topography. gentle and constant sea wave height, usually influenced by wind speed. One area of Indonesia that has these parameters is Binongko Island, Wakatobi, Southeast Sulawesi, whose wave characteristics are a basic reference for exploring the enormous energy potential of the seas and oceans in that location. The author uses a simulation model with the Graphical User Interface (GUI) method in the Matlab program to determine the potential energy produced. The data used in this research uses data analysis from the Southeast Sulawesi Regional Meteorology, Climatology, and Geophysics Agency using data samples for 2021 and 2022. The data used includes sea wave height data and sea wave period data at the research location in 2021 and the 2022 time period. Through this data, it is known that Binongko Island, Wakatobi, Southeast Sulawesi 2021 will have minimum sea wave height. of 0.5125 meters and a maximum value of 1.04167 meters, for the minimum sea wave period of 2.34993 seconds and a maximum value of 3.57008 seconds, while in 2022 the minimum sea wave height will be 0.341667 meters and the maximum value will be 0.895833 meters, for a minimum sea wave period of 1.91045 and a maximum value of 3.30481 seconds. With this data, Binongko Island, Wakatobi, Southeast Sulawesi in 2021 will be able to produce a minimum of 20.8844 watts and a maximum of 75.7116 watts, while in 2022 it will be able to produce a minimum of 12.7715 watts and a maximum of 43.5652 watts.

EFFICIENCY OPTIMIZATION OF OWC WAVE ENERGY CONVERTERS BY INCIDENT FLOW STEERING

In recent years, the interest in capturing wave energy has increased notably due to the immense potential available and the need to deepen and diversify the use of renewable energies. Oscillating Water Column (OWC) technology is one of the most technologically developed, with the world's first grid-connected demonstration plant located in Mutriku. However, the efficiency of this technology is still insufficient for its consolidation in the energy mix. Therefore, the present work presents an advance in the increase in the extraction of energy from the incident waves through the installation of a frontal ramp that directs the wave flow towards the vertical chamber of the oscillating column. The improvement achieved will be determined by means of a CFD (Computational Fluid Dynamics) 2D scaled model, which represents the interaction of the structure with different incident wave states. Different geometric options of the ramp are analyzed in comparison with the usual vertical wall option where the phenomenon of pure reflection occurs. This work aims to contribute to improving the efficiency of OWC technology, favoring the use of wave energy as a renewable energy source. Keywords: Oscillating Water Column (OWC), wave energy converter (WEC), Response Amplitude Operator (RAO), Computational Fluid Dynamics (CFD), wave energy

Investigating Soil Pore Network Connectivity in Varied Vegetation Types Using X-ray Tomography

The ecological environment in southwestern China is fragile. Due to the significant preferential flow in vertical and horizontal directions and poor water conservation ability, vegetation degradation still exists under conditions of abundant rainfall. Therefore, the pore connectivity and infiltration characteristics in shallow soil under typical local vegetation need to be studied. A calculation model for the vertical connectivity of soil macropores was independently constructed, and differences in soil macropore structures and the degree of vertical connectivity in typical vegetation types (natural secondary forest, natural grassland, Yunnan pine plantation, eucalyptus plantation, cypress plantation, mulberry bushes) were investigated by CT scanning technology of undisturbed soil columns. The results showed that the vertical connectivity of large pores in the shallow soil of the region can be quantitatively described by X-ray tomography, and the total surface area and cumulative curvature of macropores in natural grassland soil were two or three times that in artificial vegetation. The concentration area of macropores in the soil of artificial forestland was closer to the surface, and the tendency of macropore preferred path decreased by 76.18% around 30 cm depth in the soil. The vertical connection of soil macropores in artificial forests was significantly lower than that of natural secondary forestlands (33.03%) and natural grasslands (36.75%). The restoration of the plantation improved surface soil pore structure, and the vertical connectivity of soil is nearly 20% less than that of natural vegetation types (natural secondary forestland, natural grassland), which reduced water outflow rate by nearly 44% and electrolyte content by nearly 14% at a depth of 30 cm. This study provided data and research directions for the study of hydrological processes in local forest vegetation and technical support for solving the problems of soil water loss and forestland water conservation in southwestern China.