Column Volume Research Articles

From protein structure to an optimized chromatographic capture step using multiscale modeling.

Optimizing a biopharmaceutical chromatographic purification process is currently the greatest challenge during process development. A lack of process understanding calls for extensive experimental efforts in pursuit of an optimal process. In silico techniques, such as mechanistic or data driven modeling, enhance the understanding, allowing more cost-effective and time efficient process optimization. This work presents a modeling strategy integrating quantitative structure property relationship (QSPR) models and chromatographic mechanistic models (MM) to optimize a cation exchange (CEX) capture step, limiting experiments. In QSPR, structural characteristics obtained from the protein structure are used to describe physicochemical behavior. This QSPR information can be applied in MM to predict the chromatogram and optimize the entire process. To validate this approach, retention profiles of six proteins were determined experimentally from mixtures, at different pH (3.5, 4.3, 5.0, and 7.0). Four proteins at different pH's were used to train QSPR models predicting the retention volumes and characteristic charge, subsequently the equilibrium constant was determined. For an unseen protein knowing only the protein structure, the retention peak difference between the modeled and experimental peaks was 0.2% relative to the gradient length (60 column volume). Next, the CEX capture step was optimized, demonstrating a consistent result in both the experimental and QSPR-based methods. The impact of model parameter confidence on the final optimization revealed two viable process conditions, one of which is similar to the optimization achieved using experimentally obtained parameters. The multiscale modeling approach reduces the required experimental effort by identification of initial process conditions, which can be optimized.

Dynamics of femtosecond laser-driven liquid jets

The complex dynamics of flowing liquids under external stimuli can lead to significant changes in the direction of liquid jets. In this study, we utilized a femtosecond laser to drive a flowing liquid column and analyzed the effect of varying laser energies on the direction of the liquid flow. Additionally, we examined how the diameter and volume of the liquid column influenced its directional behavior when driven by the laser. By comparing the driving angles under different conditions, we explored the principles governing the femtosecond laser-driven liquid columns. These findings offer valuable insights into the field of liquid dynamics, showcasing femtosecond lasers as a good method for controlling liquid flows.

Countercurrent chromatography isolation of green propolis biomarkers: Potential blockers of SARS-COV-2 RBD and ACE2 interaction

Propolis is a natural resinous mixture produced by honeybees with numerous biological activities. Considering the recently reported potential of propolis as an adjuvant in COVID-19 treatment, a methodology for the fractionation of the hexane extract of Brazilian green propolis (HEGP) was developed for the obtention of prenylated biomarkers by countercurrent chromatography. The inhibition of the interaction between the receptor binding domain (RBD) of spike and ACE2 receptor was evaluated by the Lumitᵀᴹ immunoassay. Fractionation of HEGP was performed by both normal (CCC1 and CCC2, with extended elution) and reversed (CCC3) phase elution-extrusion modes with the solvent system hexane–ethanol–water 4:3:1. The normal elution mode of CCC1 (471 mg HEGP in a 80 mL column volume, 1.6 mm id) was scaled-up (CCC5, 1211 mg HEGP in a 112 mL column volume, 2.1 mm id), leading to the isolation of 89.9 mg of artepillin C, 1; 52.7 mg of baccharin, 2; and 26.6 mg of chromene, with purities of 93 %, 83 % and 88 %, respectively, by HPLC-PDA. Among the isolated compounds, artepillin C, 1, and baccharin, 2, presented the best results in the Lumitᵀᴹ immunoassay, showing 67% and 51% inhibition, respectively, at the concentration of 10 μM. This technique proved to be of low operational cost and excellent reproducibility.

The Relation between Variances of 3D Density and Its 2D Column Density Revisited

We revisit the relation between the variance of three-dimensional (3D) density ( σρ2 ) and that of the projected two-dimensional (2D) column density ( σΣ2 ) in turbulent media, which is of great importance in obtaining turbulence properties from observations. Earlier studies showed that σΣ/Σ02/σρ/ρ02=R , where Σ/Σ0 and ρ/ρ 0 are 2D column and 3D volume densities normalized by their mean values, respectively. The factor R depends only on the density spectrum for isotropic turbulence in a cloud that has similar dimensions along and perpendicular to the line of sight. Our major findings in this paper are as follows. First, we show that the factor R can be expressed in terms of N, the number of independent eddies along the line of sight. To be specific, σΣ/Σ02/σρ/ρ02 is proportional to ∼1/N due to the averaging effect arising from independent eddies along the line of sight. Second, we show that the factor R needs to be modified if the dimension of the cloud in the line-of-sight direction is different from that in the perpendicular direction. However, if we express σΣ/Σ02/σρ/ρ02 in terms of N, the expression remains same even in the case the cloud has different dimensions along and perpendicular to the line of sight. Third, when we plot NσΣ/Σ02 against σρ/ρ02 , two quantities roughly lie on a single curve regardless of the sonic Mach number, which implies that we can directly obtain the latter from the former. We discuss observational implications of our findings.

Investigating Temporal Characteristics of Polarimetric and Electrical Signatures in Three Severe Storms: Insights from the VORTEX-Southeast Field Campaign

Abstract The dual-polarization radar characteristics of severe storms are commonly used as indicators to estimate the size and intensity of deep convective updrafts. In this study, we track rapid fluctuations in updraft intensity and size by objectively identifying polarimetric fingerprints such as ZDR and KDP columns, which serve as proxies for mixed-phase updraft strength. We quantify the volume of ZDR and KDP columns to evaluate their utility in diagnosing temporal variability in lightning flash characteristics. Specifically, we analyze three severe storms that developed in environments with low-to-moderate instability and strong 0–6-km wind shear in northern Alabama during the 2016–17 VORTEX-Southeast field campaign. In these three cases (a tornadic supercell embedded in stratiform precipitation, a nontornadic supercell, and a supercell embedded within a quasi-linear convective system), we find that the volume of the KDP columns exhibits a stronger correlation with the total flash rate. The higher covariability of the KDP column volume with the total flash rate suggests that the overall electrification and precipitation microphysics were dominated by cold cloud processes. The lower covariability with the ZDR column volume indicates the presence of nonsteady updrafts or a less prominent role of warm rain processes in graupel growth and subsequent electrification. Furthermore, we observe that the majority of cloud-to-ground (CG) lightning strikes a carried negative charge to the ground. In contrast to findings from a tornadic supercell over the Great Plains, lightning flash initiations in the Alabama storms primarily occurred outside the footprint of the ZDR and KDP column objects. Significance Statement This study quantifies the correlation between mixed-phase updraft intensity and total lightning flash rate in three severe storms in northern Alabama. In the absence of direct updraft velocity measurements, we use polarimetric signatures, such as ZDR and KDP columns, as proxies for updraft strength. Our analysis of polarimetric radar and lightning mapping array data reveals that the lightning flash rate is more highly correlated with the KDP column volume than with the ZDR column volume in all three storms examined. This contrasts with previous findings in storms over the central Great Plains, where the ZDR column volume showed higher covariability with flash rate. Interestingly, lightning initiation in the Alabama storms mainly occurred outside the ZDR and KDP column areas, contrary to previous findings.

Mass spectrometry coupling of chip-based supercritical fluid chromatography enabled by make-up flow-assisted backpressure regulation

This work introduces a novel microfluidic backpressure pressure control developed for chip-based supercritical fluid chromatography (chipSFC). The presented on-chip pressure control mechanism involves the post-column addition of a viscous make-up stream, which enables pressure regulation within the range of 73 to 130 bar range. In contrast to approaches using mechanical backpressure regulators, this chip-based make-up-assisted pressure regulation offers a wear-free alternative that functions entirely through fluidic means and contributes minimally to extra column volume. It prevents phase separation of the supercritical mobile phase and, therefore, expands the analytical scope of chipSFC to detection systems with an ambient pressure interface. This was demonstrated by a proof-of-principle experiment, where a model mixture was separated within 30 s and detected using atmospheric pressure ionisation mass spectrometry.Graphical abstract

Removal of microplastics from agricultural runoff using biochar: a column feasibility study

Plastics are extensively used in agriculture, but their weathering and degradation generates microplastics (MPs) that can be carried by runoff into water bodies where they can accumulate and impact wildlife. Due to its physicochemical properties, biochar has shown promise in mitigating contaminants in agricultural runoff. However, few studies have examined its effectiveness at removing MPs. In this study, we assessed MP pollution (>30 μm) in runoff from a farm in the Mississippi Delta and examined the effectiveness of biochar (pinewood and sugarcane) to remove MPs from aqueous solutions. Using micro-Fourier Transform Infrared spectroscopy (µ-FTIR), we observed an average of 237 MPs/L (range 27–609) in the runoff, with most particles identified as polyethylene, polyamide, polyvinyl chloride, polyurethane, acrylonitrile butadiene styrene, and polyarylamide. Biochar columns effectively removed MPs from runoff samples with reductions ranging from 86.6% to 92.6%. MPs of different sizes, shapes, and types were stained with Nile red dye (to facilitate observation by fluorescence) and quantified their downward progress with multiple column volumes of water and wet/dry cycles. Smaller MPs penetrated the columns further, but ≥90% of MPs were retained in the ∼20 cm columns regardless of their shape, size, and type. We attribute these results to physical entrapment, hydrophobic behaviors, and electrostatic interactions. Overall, this proof-of-concept work suggests biochar may serve as a cost-effective approach to remove MPs from runoff, and that subsequent field studies are warranted.

Grafting chromatographic monoliths with charged linear polymers for highly productive and selective protein or plasmid DNA purification

Increased requirements for plasmid DNA (pDNA) in gene therapy and vaccination efforts bring the need for efficient large-scale production processes with high purity and productivity. Chromatographic purification of pDNA is often a bottleneck due to low dynamic binding capacities (DBC), pDNA recovery and/or low selectivity of columns. Ion exchangers (IEX) with charged groups on surface extenders (polyions formed by a grafting reaction and subsequents modification) have attracted attention due to their greatly enhanced productivity compared with traditional IEX. The ultimate goal of our investigation was to increase the DBC for pDNA on a monolith stationary phase by grafting the surface with linear polymethacrylate brushes (grafted chains), while retaining high pDNA recovery and chromatographic selectivity between pDNA and RNA impurities. The inner walls of the Convective Interaction Media® monolith channels were successfully coated with polyglycidyl methacrylate linear chains of varying densities and lengths by employing a controlled radical polymerization reaction. Glycidyl groups throughout the length of the formed grafted chains were converted to cation- (CEX) or anion-exchanging (AEX) moieties. Preliminary chromatographic evaluation of novel grafted CEX columns for different model proteins exhibited 10-times higher DBC with high elution recovery (99 %). Similar DBC improvement was proven for 7.3 kilobase pair pDNA, where the DBC10 increased from 1.8 mg pDNA per mL of non-grafted chromatographic support up to 15.2 mg per the most densely distributed and the longest grafted chains, modified with weak AEX groups. However, the DBC increase was partially at the expense of pDNA elution recovery, which presumably decreased due to entanglement of pDNA molecules inside the dense grafted layer. To confirm the hypothesis, we have prepared a series of columns differing in length and density of grafted chains and tested them for pDNA capture. A satisfactory compromise between high pDNA DBC and elution recovery was found with relatively long and low density grafted chains. The optimal grafted AEX column with 1 mL bed volume was evaluated for pDNA capture from neutralized E. coli lysate. With a capacity of 13.5 mg pDNA per mL support, ≥95 % elution recovery, and complete RNA clearance, the pDNA was successfully purified at loading flow rate of at least 15 column volumes per min. According to our knowledge and literature search, these process characteristics would enable one of the highest pDNA isolation productivities to be achieved with currently tested IEX.

Rapid purification and scale-up of tilianin using counter-current chromatography with rectangular horizontal tubing

In counter-current chromatography (CCC), linear scale-up is an ideal amplification strategy. However, when transferring from analytical to predictable preparative processes with high throughput, linear scale-up would be challenging due to limitations imposed by differences in instrument parameters, such as gravitational forces, tubing cross-section area, tubing length, column volume and flow rate. Some effective scale-up strategies have been studied for different instrument parameters, but so far, these scale-up works have only been tested on standard circular (SC) tubing.The previous research of our group found that rectangular horizontal (RH) tubing can double the separation efficiency compared with conventional SC tubing, and has industrial production potential. This paper used the separation of tilianin from Dracocephalum moldavica L. as an example to demonstrate how to scale up the optimized process from analytical SC tubing to preparative RH tubing. After systematic optimization of solvent systems, sample concentration and flow rate on the analytical CCC, the optimized parameters obtained were successfully transferred to the preparative CCC. The results showed that a crude sample of 2.07 g was successfully separated using a solvent system of n-hexane - ethyl acetate - ethanol - water (1:4:1:5, v/v/v/v) in reversed phase mode, and the three consecutive separations produced a total of 380 mg tilianin in 75 min with high purities of 98.3%, as analyzed by HPLC. The total throughput achieved from the analytical to semi-preparative scale was improved by 138 times (from 12 mg/h to 1.66 g/h), while the column volume was increased by only 46.5 times (from 15.5 mL to 720 mL). This is the successful application of CCC for the separation and purification of tilianin. Given that SC tubing is the traditional configuration for CCC columns, this study is a necessary step to prove the applicability of RH tubing columns for routine use and potential large-scale industrial applications.

Ground-based MAX-DOAS observations of formaldehyde and glyoxal in Xishuangbanna, China

Multi-axial differential optical absorption spectroscopy (MAX-DOAS) measurements were conducted in Xishuangbanna, Yunnan, China, between November 1, 2021 and June 30, 2022 to obtain vertical distributions of formaldehyde (HCHO) and glyoxal (CHOCHO). The observations show an increase in vertical column densities (VCDs) and volume mixing ratios (VMRs) for both HCHO and CHOCHO concentrations during periods of biomass combustion. The VCDs of HCHO and CHOCHO from TROPOMI are in good agreement with the MAX-DOAS observations. (R2HCHO = 0.71; R2CHOCHO = 0.70). Regarding seasonal variations, HCHO predominantly occupies the upper layer (400-800 m) during the biomass burning, possibly attributed to the formation of secondary HCHO as the plume ascends during combustion. CHOCHO is primarily found in the lower layer (0-200 m), suggesting a longer lifespan for HCHO compared to CHOCHO, preventing the latter from diffusing to higher altitudes. Concerning the daily variation patterns, both HCHO and CHOCHO VMRs exhibited peaks at 9:00 and 13:00, which were attributed to the nighttime accumulation and midday oxidation. Furthermore, we also investigated the sources of volatile organic compounds (VOCs) using the CHOCHO to HCHO ratio (RGF). During the period of biomass burning, there are minimal differences in the daily RGF across layers, indicating that biomass burning is the predominant source. During the non-biomass burning period, the daily RGF shows significant differences among layers, indicating that emissions from biological and anthropogenic sources primarily contribute during the period.

Purification of Anti-Mycobacterium tuberculosis MPT64 Immunoglobulin-Y from Egg-Yolk Supernatant Using Thiophilic Adsorption Chromatography

The significance of immunoglobulin Y (IgY) as a particular antibody equal to mammalian IgG is well understood. However, due to a lack of reliable purification procedures, producing highly pure IgY remains problematic. In this study, we aimed to optimize the recovery of pure IgY anti-MPT64 using thiophilic adsorption chromatography. The purification of IgY anti-MPT64 was achieved by initial PEG lipid precipitation, then an optimized purification by varying the gradient concentration of elution buffer into five steps gradient (0-20, 20-40, 40-60, 60-80, and 80-100%v/v) for three injection column volume (CV) each and two steps gradient (0-50 and 50-100%v/v) in eight CV for each concentration. The obtained IgY was characterized by SDS-PAGE and dot-blot then determined its content levels using the Lowry method. The results showed that the five steps gradient purification was found to provide a better purity level of IgY than the two steps gradient. However, the IgY content obtained in the two steps gradient purification (2.2632± 0.011 mg/mL) was higher than the five steps gradient purification (1.35482 ± 0.023 mg/mL). Nevertheless, both purified IgY results can recognize MPT64 protein through a dot blot test. Therefore, it can be summarized that thiophilic adsorption chromatography with five steps gradient of purification was an efficient process to obtain a higher purity of IgY anti-MPT64, especially to be targeted as a diagnostic kit component for MPT64 detection.

Adaptable SEC-SAXS data collection for higher quality structure analysis in solution.

The two major challenges in synchrotron size-exclusion chromatography coupled in-line with small-angle x-ray scattering (SEC-SAXS) experiments are the overlapping peaks in the elution profile and the fouling of radiation-damaged materials on the walls of the sample cell. In recent years, many post-experimental analyses techniques have been developed and applied to extract scattering profiles from these problematic SEC-SAXS data. Here, we present three modes of data collection at the BioSAXS Beamline 4-2 of the Stanford Synchrotron Radiation Lightsource (SSRL BL4-2). The first mode, the High-Resolution mode, enables SEC-SAXS data collection with excellent sample separation and virtually no additional peak broadening from the UHPLC UV detector to the x-ray position by taking advantage of the low system dispersion of the UHPLC. The small bed volume of the analytical SEC column minimizes sample dilution in the column and facilitates data collection at higher sample concentrations with excellent sample economy equal to or even less than that of the conventional equilibrium SAXS method. Radiation damage problems during SEC-SAXS data collection are evaded by additional cleaning of the sample cell after buffer data collection and avoidance of unnecessary exposures through the use of the x-ray shutter control options, allowing sample data collection with a clean sample cell. Therefore, accurate background subtraction can be performed at a level equivalent to the conventional equilibrium SAXS method without requiring baseline correction, thereby leading to more reliable downstream structural analysis and quicker access to new science. The two other data collection modes, the High-Throughput mode and the Co-Flow mode, add agility to the planning and execution of experiments to efficiently achieve the user's scientific objectives at the SSRL BL4-2.

Enhancing the purification of crocin-I from saffron through the combination of multicolumn countercurrent chromatography and green solvents

Crocin-I, a valuable natural compound found in saffron (Crocus sativus L.), is the most abundant among the various crocin structures. Developing a cost-effective and scalable purification process to produce high-purity crocin-I is of great interest for future investigations into its biological properties and its potential applications in the treatment of neurological disorders. However purifying crocin-I through single-column preparative chromatography (batch) poses a yield-purity trade-off due to structural similarities among crocins, meaning that the choice of the collection window sacrifices either yield in benefit of higher purity or vice versa. This study demonstrates how the continuous countercurrent operating mode resolves this dilemma. Herein, a twin-column MCSGP (multicolumn countercurrent solvent gradient purification) process was employed to purify crocin-I. This study involved an environmentally friendly ethanolic extraction of saffron stigma, followed by an investigation into the stability of the crocin-I within the feed under varying storage conditions to ensure a stable feed composition during the purification. Then, the batch purification process was initially designed, optimized, and subsequently followed by the scale-up to the MCSGP process. To ensure a fair comparison, both processes were evaluated under similar conditions (e.g., similar total column volume). The results showed that, at a purity grade of 99.7%, the MCSGP technique demonstrated significant results, namely + 334% increase in recovery + 307% increase in productivity, and − 92% reduction in solvent consumption. To make the purification process even greener, the only organic solvent employed was ethanol, without the addition of any additive. In conclusion, this study presents the MCSGP as a reliable, simple, and economical technique for purifying crocin-I from saffron extract, demonstrating for the first time that it can be effectively applied as a powerful approach for process intensification in the purification of natural products from complex matrices.Graphical

Hydroxyalkyl Amination of Agarose Gels Improves Adsorption of Bisphenol A and Diclofenac from Water: Conceivable Prospects

The hydroxyalkyl amination of agarose gels was studied as an approach to improve adsorption of polyphenols and pharmaceuticals from water. Three commercially available agarose gels, Zetarose FlashFlow4, ZetaCell-CL6B and Sepharose 4B were chemically modified using tris-(hydroxymethyl)aminomethane, TRIS, and ethanolamine, EA. The adsorbed amounts of bisphenol A and diclofenac were significantly higher on TRIS- and EA-derivatives compared with the parent gels. Regarding bisphenol A adsorption on TRIS-ZetaCell-CL6B, a maximal adsorption capacity, Q max of 16 μmol/mL gel and an equilibrium dissociation constant KL of 2.7 × 10−4 mol/L were observed. Filtration of diclofenac-contaminated water through TRIS-Zetarose FlashFlow 4 resulted in a 10-fold reduction of the pollutant concentration within 64 column volumes of the effluent. The moderate binding affinity of polyphenols to TRIS- and EA-adsorbents facilitates efficient polyphenol desorption and column regeneration. The effects of TRIS- and EA-substituents in agarose gels, can be harnessed for the development of environmental adsorbents, as well as for the preparative separation of polyphenols and pharmaceuticals. We consider the physical shapes and textures of the prospective adsorbents with a particular focus on spongy macroporous cryogels. These innovative materials hold promise for future applications in liquid and air filtration.

Volume Density Structure of the Central Molecular Zone NGC 253 through ALCHEMI Excitation Analysis

We present a spatially resolved excitation analysis for the central molecular zone (CMZ) of the starburst galaxy NGC 253 using the data from the Atacama Large Millimeter/submillimeter Array Comprehensive High-resolution Extragalactic Molecular Inventory, whereby we explore parameters distinguishing NGC 253 from the quiescent Milky Way’s Galactic center (GC). Non-LTE analyses employing a hierarchical Bayesian framework are applied to Band 3–7 transitions from nine molecular species to delineate the position–position–velocity distributions of column density ( NH2 ), volume density ( nH2 ), and temperature (T kin) at 27 pc resolution. Two distinct components are detected: a low-density component with (nH2,Tkin)∼(103.3cm−3,85K) and a high-density component with (nH2,Tkin)∼(104.4cm−3,110K) , separated at nH2∼103.8cm−3 . NGC 253 has ∼10 times the high-density gas mass and ∼3 times the dense-gas mass fraction of the GC. These properties are consistent with their HCN/CO ratio but cannot alone explain the factor of ∼30 difference in their star formation efficiencies (SFEs), contradicting the dense-gas mass to star formation rate scaling law. The nH2 histogram toward NGC 253 exhibits a shallow declining slope up to nH2∼106cm−3 , while that of the GC steeply drops in nH2≳104.5cm−3 and vanishes at 105 cm−3. Their dense-gas mass fraction ratio becomes consistent with their SFEs when the threshold nH2 for the dense gas is taken at ∼104.2−4.6 cm−3. The rich abundance of gas above this density range in the NGC 253 CMZ, or its scarcity in the GC, is likely to be the critical difference characterizing the contrasting star formation in the centers of the two galaxies.

Analysis and reflection on the teaching of column volumes based on aesthetic education

Analysis and reflection on the teaching of column volumes based on aesthetic education

BART: A transferable liquid chromatography retention time library for bile acids

Identification of unknown bile acids, especially the distinguishment between isomers, requires retention times of a large number of reference standards, which are often not commercially available. Meanwhile, published retention information cannot be directly transferred across labs due to the differences between liquid chromatography (LC) systems, such as different extra column volume and dwell volume. To improve this situation, a transferrable retention time library for bile acids named BART was developed. BART was composed of isocratic retention models of 272 bile acids and a software tool to predict their gradient retention times on various LC systems. The isocratic retention times of bile acids were acquired on a Waters BEH C18 column with mobile phases of acidic ammonium acetate buffer and acetonitrile, and fit to the quadratic solvent strength model (QSSM). Segmented linear gradient retention times were calculated with holdup time (t0), dwell time (tD) and actual gradient profile corrected using 21 bile acid calibration standards. In addition to the reference system where the isocratic retention times were acquired, this approach has been validated on four other LC-MS systems in four labs with two gradient methods. Average root mean square errors (RMSE) between predicted and experimental retention times were 0.052 and 0.054 min for the two gradients tested, which were 9-fold more accurate than referring to a static retention time library. The library is freely available at https://bafinder.github.io/.

CO2 capture from flue gas using EAE-1DMA2P solvent in a packed column

The mass transfer plays a decisive role in screening and evaluation of amine solvents as well as in designing and scale-up of CO2 capture. The influence of different operating parameters on mass transfer processes in CO2 capture by EAE-1DMA2P amine solution was thoroughly investigated using a small-scale packed column. The mass transfer mechanism of EAE-1DMA2P amine solution in the capture of CO2 was discussed by determining both mass transfer control steps and the reaction region between CO2 and amine solvent, which provided a theoretical guidance for subsequent development of new solvents for large-scale applications. In addition, based on the experimental observations of KGav, a prediction model for KGav of EAE-1DMA2P composite amine solution was established by comparing two empirical models viz., Kohl-Riesenfeld-Astarita and Sheng of which Kohl-Riesenfeld-Astarita model provided a better fitting of the experimental results. The commonly used amine solvents and EAE-1DMA2P composite solvents were further compared and analyzed in terms of packed column size design and solvent consumption. The required packed column volume using EAE-1DMA2P composite solvent was only 4.95 m3, while the EAE-1DMA2P composite solvent has the smallest solvent consumption among all the solvents, indicating that this solvent has a lower operating cost than the other solvents, suggesting better industrial application.

Maxillary sinus volume and adenoid tissue size in children: computed tomographic evaluation

Abstract OBJECTIVES. Adenoid hypertrophy is related to obstructive sleep apnea syndrome (OSAS). In this study, we investigated the relationship between adenoid tissue size and maxillary sinus volumes by examining the paranasal sinus computed tomography (PNSCT) images. MATERIAL AND METHODS. PNSCT images of 41 children were included in this study. According to the adenoid/nasopharynx (A/NP) ratio, two groups were identified. Group 1 (n=19, 9 males and 10 females): A/NP≤0.50 and Group 2 (n=22, 16 males and 6 females): A/NP>0.50. The following measurements were performed: adenoid width, NP width, A/NP ratio, air column, soft palate, maxillary sinus volume and mucosal thickening. RESULTS. Adenoid width of group 2 (16.11±2.26 mm) was higher than that of group 1 (10.97±2.26 mm). In group 2, with an A/NP ratio of 0.61±0.07, the air column value (6.30±2.27 mm) was lower than that of group 1 with an A/NP ratio of 0.40±0.07 (air column value: 10.06±3.78 mm). NP width, soft palate and maxillary sinus volumes were not different between groups 1 and 2. As the adenoid width increased, the A/NP ratio increased, and the air column decreased. CONCLUSION. The increase of the A/NP ratio was related to the decreased air column. In older children, air column and bilateral maxillary sinus volumes increased. In males, the adenoid width value was larger than that in females. There was no significant relationship between A/NP ratio and maxillary sinus volume or NP width values. However, higher A/NP ratio values were related to lower air column measurements.

Model based process optimization of an industrial chromatographic process for separation of lactoferrin from bovine milk

Model based process development using predictive mechanistic models is a powerful tool for in-silico downstream process development. It allows to obtain a thorough understanding of the process reducing experimental effort. While in pharma industry, mechanistic modeling becomes more common in the last years, it is rarely applied in food industry. This case study investigates risk ranking and possible optimization of the industrial process of purifying lactoferrin from bovine milk using SP Sepharose Big Beads with a resin particle diameter of 200 µm, based on a minimal number of lab-scale experiments combining traditional scale-down experiments with mechanistic modeling. Depending on the location and season, process water pH and the composition of raw milk can vary, posing a challenge for highly efficient process development.A predictive model based on the general rate model with steric mass action binding, extended for pH dependence, was calibrated to describe the elution behavior of lactoferrin and main impurities. The gained model was evaluated against changes in flow rate, step elution conditions, and higher loading and showed excellent agreement with the observed experimental data. The model was then used to investigate the critical process parameters, such as water pH, conductivity of elution steps, and flow rate, on process performance and purity. It was found that the elution behavior of lactoferrin is relatively consistent over the pH range of 5.5 to 7.6, while the elution behavior of the main impurities varies greatly with elution pH. As a result, a significant loss in lactoferrin is unavoidable to achieve desired purities at pH levels below pH 6.0. Optimal process parameters were identified to reduce water and salt consumption and increase purity, depending on water pH and raw milk composition. The optimal conductivity for impurity removal in a low conductivity elution step was found to be 43 mS/cm, while a conductivity of 95 mS/cm leads to the lowest overall salt usage during lactoferrin elution. Further increasing the conductivity during lactoferrin elution can only slightly lower the elution volume thus can also lead to higher total salt usage. Low flow rates during elution of 0.2 column volume per minute are beneficial compared to higher flow rates of 1 column volume per minute.The, on lab-scale, calibrated model allows predicting elution volume and impurity removal for large-scale experiments in a commercial plant processing over 106 liters of milk per day. The successful model extrapolation was possible without recalibration or detailed knowledge of the manufacturing plant. This study therefore provides a possible pathway for rapid process development of chromatographic purification in the food industries combining traditional scale-down experiments with mechanistic modeling.