Hold-up Volume Research Articles

Impact on Quality during In-Use Preparation of an Antibody Drug Conjugate with Eight Different Closed System Transfer Device Brands.

The aim of this study was to investigate the in-use compatibility of eight commercially available closed system transfer device brands (CSTDs) with a formulated model antibody drug conjugate (ADC). Overall, in-use simulated dosing preparation applying the CSTD systems investigated raised concerns for several product quality attributes. The incompatibilities observed were mainly associated with increased visible and subvisible particles formation as well as significant changes in holdup volumes. Visible and subvisible particles contained heterogeneous mixtures of particle classes, with the majority of subvisible particles associated with silicone oil leaching from CSTD systems during simulated dose preparation upon contact with the ADC formulation. These observations demonstrate that CSTD use may adversely impact product quality and delivered dose which could potentially lead to safety and efficacy concerns during administration. Other product quality attributes measured including turbidity, color, ADC recovery, and purity by size exclusion HPLC, did not show relevant changes. It is therefore strongly recommended to test and screen the compatibility of CSTDs with the respective ADC, in a representative in-use simulated administration setting, during early CMC development, i.e., well before the start of clinical studies, to include information about compatibility and to ensure that the CSTD listed in the manuals of preparation for clinical handling has been thoroughly assessed before human use.

Improved sieving coefficient in perfusion cell culture with reduced effective filtration length of hollow fibers.

The hollow fiber filter is the primary cell-retention device used in high-density perfusion cell culture and often used in an alternating tangential flow (ATF) configuration. The limited commercially available diaphragm pumps for ATF prevent utilization of vertical space when scaling beyond 500 L. Stacking hollow fiber filters coupled with viscous cell culture imposes vacuum pressure exceeding facility capabilities. Additionally, the longer filter assembly increases the hold-up volume and exceeds the diaphragm pump's fluid exchange capacity. The conventional tangential flow filtration (TFF) configuration circumvents this issue by exchanging culture from the bioreactor and cell-retention device in a unidirectional recirculation loop; however, the increased filter length when scaled up exacerbates the TFF's inherent issue with product retention from Starling flow. Stacking commercially available 20 cm TFF filters to make up the similar single-module length TFF used for the platform 3 and 50 L perfusion process at 41.5 and 65 cm, respectively, attempts to reduce fouling caused by Starling flow. The permeate of a single-module filter is partitioned into short independent segments through serially stacked filters, each harvested separately. By partitioning the permeate, the sieving coefficient increased for both 3 and 50 L scales. Reduction of Starling flow was confirmed with lower total hydraulic membrane resistance throughout the culture. This work demonstrates a method for increasing sieving coefficient and filter capacity by stacking TFF filters with independent permeate streams.

Impact of Closed System Transfer Device (CSTD) Handling Procedure for Low-Transfer-Volume Dose Preparation of Biologic Drug Products

Many challenges have been identified for ensuring compatibility of closed system transfer devices (CSTDs) with biologic drug products. One challenge is large hold-up volumes (HUVs) of CSTD components, which can be especially problematic with early-stage biologics when low transfer volumes smaller than the nominal fill volume may be used to achieve a wide range of doses with a single drug product configuration. Here, we identified possible CSTD handling techniques during dose preparation of a drug product requiring small volume transfers during reconstitution, intermediate dilution, and dilution in an IV bag, and systematically evaluated the impact of these handling procedures on the ability to deliver an accurate dose to the next step. We show that small changes to CSTD procedures can have a major impact on dose accuracy, depending on both CSTD HUVs and drug product-specific transfer volumes. We demonstrate that it is possible to craft CSTD instructions for use to mitigate these issues, and that the dose accuracy for specific drug product/CSTD combinations can be estimated using theoretical equations. Finally, we explored potential downsides of these mitigations. Our results emphasize key factors for consideration by both drug and CSTD manufacturers when assessing compatibility and providing CSTD instructions for use with biologics requiring low transfer volumes during dose preparation.

Comprehensive analysis of the effective and intra-particle diffusion of weakly retained compounds in silica hydrophilic interaction liquid chromatography columns

A detailed analysis of intra-particle volumes and layer thicknesses and their effect on the diffusion of solutes in hydrophilic interaction liquid chromatography (HILIC) was made. Pycnometric measurements and the retention volume of deuterated mobile phase constituents (water and acetonitrile) were used to estimate the void volume inside the column, including not only the volume of the mobile phase but also part of the enriched water solvent acting as the stationary phase in HILIC. The mobile phase (hold-up) volume accessible to non-retained components was estimated using a homologous series approach. The joint analysis of the different approaches indicated the formation of enriched water layers on the hydrophobic silica mesopore walls with a thickness varying significantly with mobile phase composition. The maximal thickness of the enriched water layers, which corresponded to the minimum void volume accessible to unretained solutes, marked a transition in the retention behavior of the studied analytes. Discrepancies between deuterated solvent measurements and pycnometry were explained by the existence of an irreplaceable water layer adsorbed on the silica surface. Regarding the diffusion behavior in HILIC, peak parking experiments were used to interpret the influence of the acetonitrile content on the effective diffusion coefficient Deff. A systematic decrease in Deff and molecular diffusion Dm was observed with decreasing acetonitrile concentration, primarily attributed to variations in mobile phase viscosity. Notably, Deff/Dm remained nearly unaffected by variations in mobile phase composition. Finally, the effective medium theory was used to make a comprehensive analysis of Dpart/Dm to study the contribution to band broadening when the solute resides in the mesopores. The obtained data unveiled a curvature with a minimum corresponding to conditions of maximum water-layer thickness and retention. For the weakly retained compounds (k’ < 0.5) the Dpart/Dm-values were found to be relatively high (order of 0.35-0.5), which directly reflects the high γsDs/Dm-values that were observed (order 0.35-7).

Characterization of solute-solvent interactions in liquid chromatography systems: A fast method based on Abraham's linear solvation energy relationships

The Abraham's solvation parameter model, based on linear solvation energy relationships (LSER), allows the accurate characterization of the selectivity of chromatographic systems according to solute-solvent interactions (polarizability, dipolarity, hydrogen bonding, and cavity formation). However, this method, based on multilinear regression analysis, requires the measurement of the retention factors of a considerably high number of compounds, turning it into a time-consuming low throughput method. Simpler methods such as Tanaka's scheme are preferred. In the present work, the Abraham's model is revisited to develop a fast and reliable method, similar to the one proposed by Tanaka, for the characterization of columns employed in reversed-phase liquid chromatography and particularly in hydrophilic interaction liquid chromatography. For this purpose, pairs of compounds are carefully selected in order to have in common all molecular descriptors except for a specific one (for instance, similar molecular volume, dipolarity, polarizability, and hydrogen bonding basicity features, but different hydrogen bonding acidity). Thus, the selectivity factor of a single pair of test compounds can provide information regarding the extent of the dissimilar solute-solvent interactions and their influence on chromatographic retention. The proposed characterization method includes the determination of the column hold-up volume and Abraham's cavity term by means of the injection of four alkyl ketone homologues. Therefore, five chromatographic runs in a reversed-phase column (four pairs of test solutes and a mixture of four homologues) are enough to characterize the selectivity of a chromatographic system. Tanaka's method is also analyzed from the LSER point of view.

Prediction of pressure gradient and hold-up in horizontal liquid-liquid pipe flow

This paper aims to propose correlations to predict pressure gradient, friction factor and fluid phase hold-up in liquid-liquid horizontal pipe flow. To develop the correlations, experiments are conducted using high viscous oils (202 and 630 mPa⋅s) in a steel pipe of length 11.25 m and length-to-diameter ratio of 708. In addition, the experimental data from the literature comprising wide range of flow and fluid properties is analyzed. For the analysis, the liquid-liquid pipe flow data is categorized into two as: stratified and dispersed. The existing friction factor correlations are modified to incorporate the effects of viscosity of the oil phase, interfacial curvature (contact/wetting angle-in lieu of material of the pipe) and fluid phase fraction. In the two-fluid model of stratified flow, the wall stress and interfacial stress correlations are substituted with superficial velocities of fluids and superficial Reynolds numbers of fluid phases replacing fluid phase velocities and fluid Reynolds numbers. Similarly, for dispersed flow, an effective Reynolds number is described as the sum of superficial Reynolds number of oil and water phases. Substituting the generally employed mean or mixture Reynolds number with the effective Reynolds number into the existing single-phase turbulent flow friction factor correlation, an effective friction factor for oil-water flow is proposed. Employing the proposed correlations, the pressure gradient across the oil-water flow and hold-up volume fraction are predicted with significant reduction in error compared with that of conventionally employed correlations. The average error and standard deviation values of −7.06%, 20.72% and 0.31%, 18.79% are found for stratified flow and dispersed flow respectively.

Understanding the impact of reactive holdup on process intensification in the design of reactive distillation column

The commercial viability of reactive distillation as a front-runner industrial process intensification technique is limited by slow reactions. It is because the diameter established by the vapor-loading method restricts large holdup volumes (or catalyst amount) to accomplish the essential conversion. However, placing a large amount of catalyst on column trays necessitates either a high tray weir height (limited to excessive column pressure drop) or a large column diameter. This study aims to investigate an alternative design approach by increasing the column diameter beyond that required for vapor loading to retain a large holdup volume. Several combinations of tray weir heights and column diameters were studied and demonstrated through case studies for three industrial processes, and their optimal designs have been reported. High catalyst holdup volume enhanced energy efficiency and overcame hydraulic limitations despite requiring large diameter vessels. These design configurations with optimized catalyst holdup also resulted in improved process economics. Fabricating a wide column diameter is a good conservative engineering procedure considering safety aspects and better design control.

Evaluation of Hold-Up Volume Determination Methods and Markers in Hydrophilic Interaction Liquid Chromatography.

Common methods for hold-up time and volume determination in Reversed-Phase Liquid Chromatography (RPLC) have been tested for Hydrophilic Interaction Liquid Chromatography (HILIC). A zwitterionic ZIC-HILIC column has been used for the testing. The pycnometric determination method, based on differences in column weight when filled with water or organic solvent, provides the overall volume of solvent inside the column. This includes the volume of eluent semi-sorbed on the packing of the column, which acts as the main stationary phase. The homologous series approach, based on the retention behavior of homologues in relation to their molecular volume, allows the determination of accurate hold-up volumes. However, the application of this method is time-consuming. In some cases, large neutral markers with poor dipolarity/polarizability and hydrogen bonding interactions can be used as hold-up volume markers. This is the case of dodecylbenzene and nonadecane-2-one in clearly HILIC behaving chromatographic systems, the use of decanophenone as a marker can be even extended to the boundary between HILIC and RPLC. The elution volume of the marker remains nearly unaffected by the concentration of ammonium acetate in the mobile phase up to 20 mM. The injection of pure solvents to produce minor base-line disturbance as hold-up markers is strongly discouraged, since solvent peaks are complex to interpret and depend on the ionic strength of the eluent.

Harmonization of Experimental Methods Used to Measure the True Hold-Up Volume of Liquid Chromatography Columns

There are as many measurement values of the true column hold-up volume, V0, as techniques applied to evaluate this most important property in liquid chromatography (LC). The relative errors made on V0 measurements using conventional “non-retained” markers—such as acetone, uracil, or thiourea in reversed-phase liquid chromatography (RPLC), or benzene or acenaphthene in hydrophilic interaction chromatography (HILIC)—can be as large as ±30%. This situation is extremely confusing for LC users who wish to classify and predict the retention behavior of LC columns. In this work, along with advances in mass spectrometry (MS) instrumentation, the hold-up volume of any LC column—including, but not limited to, RPLC, HILIC, ion exchange chromatography (IEX), and mixed-mode columns—is accurately measured by injecting labeled deuterated acetonitrile (CD3CN) molecules and detecting them selectively by MS-single ion reaction (m/z = 45) using non-labeled and pure acetonitrile (CH3CN) as the eluent. This proposed harmonization of all conventional V0 measurement methods is illustrated and successfully applied to RPLC, HILIC, anion exchange (AEX), and RP-AEX mixed-mode chromatography, irrespective of the mobile phase composition selected.

Effects of the Structure, Operation, and Physical Parameters on the Actual Phase Ratio and Interface Radius in the Separation Zone of an Annular Centrifugal Contactor

The annular centrifugal contactor (ACC) is highly efficient solvent extraction equipment and has been successfully used in many industrial fields. The phase distribution of two phases in the separation zone of the rotor is an important factor affecting the hydraulic performance of the ACC. It can be characterized by the actual phase ratio (Ro/a) and interface radius (ri). So far, there have been few systematic research studies on the effects of the structure, operation, and physical parameters on them. This work systematically investigated the effects of the structure [the radius of the heavy phase weir (ra*) and radius of the light phase weir (ro*)], operation [rotor speed (N), flow ratio (Fo/a), and total flow (Q)], and physical parameters [density difference (ρa – ρo), viscosity ratio (μo/μa), and interfacial tension (γ/γ*, γ* is the constant interfacial tension)] on the hold-up volume, Ro/a, and ri in the 20 mm ACC using a liquid fast separating method. The results show that Ro/a and ri increased with the increase of N, Fo/a, and ra* and decreased with the increase of ro*, Q, and ρa – ρo. However, μo/μa and γ/γ* have little effect on Ro/a and ri. The N can reach more than 5000 rpm when the ρa – ρo is greater than 0.2200 g/cm3. The closer the Ro/a is to 0.45, the better the hydrodynamic properties of the extracted system are in the ACC. A new model associated with these parameters was established to calculate ri. The results show that the ρa – ρo of the extraction system is the main factor affecting the ri with a degree of influence of 0.5654, followed by the combination of different-size phase weirs with a degree of influence of 0.4052. Moreover, the ri values obtained by the model are in good agreement with those obtained by experiments. The experimental results will provide relevant references for the improvement and optimization of the ACC.

Microbubbles, oscillating flow, and mass transfer coefficients in air-water bubble columns

Mass transfer coefficient kLa determines the flux of dissolved oxygen DO in bubble columns. The kLa values can be determined experimentally or predicted using a correlation. Common correlations link kLa with superficial air velocity, air hold-up volume, bubble contact time, bubble rising velocity, or bubble Sauter diameter.We compared experimentally found and predicted kLa values to evidence that none of the above correlations is applicable for microbubbles flow. The only way to accurately predict the flow is by using mass transport models needed to calculate kL. The transition from kL to kLa requires a redefinition of the specific interfacial area a for microbubbles.Microbubbles distribute DO quicker than macrobubbles. Oscillating flow can raise the DO level to saturation. The generation of oscillating flow does not require extra energy turning the subsurface aeration more energy-efficient than the surface aeration.

Modeling the Impact of Holdup Volume from Chromatographic Workstations on Ion-Exchange Chromatography

Modeling the Impact of Holdup Volume from Chromatographic Workstations on Ion-Exchange Chromatography

Liquid Hold-up Volume and Phase Ratio (A/O) of HNO3–DtBuCH18C6/n-Octanol System in an Annular Centrifugal Extractor

ABSTRACT The crown ether strontium extraction (CESE) process has been developed for the recovery or removal of Sr from acidic high-level liquid waste (HLLW) in China, where the extractant is 4’,4”(5”)-di-(tert-butyldicyclohexano)-18-crown-6 (DtBuCH18C6) and the diluent is n-octanol. When the CESE process was operated in annular centrifugal extractors (ACEs) for demonstration by cold and hot tests, it was found that the outlet aqueous phases had organic-phase entrainment even though ACEs operated at a low rotor speed. For the successful operation of the CESE process in ACEs, effects of the operating and geometrical parameters on the liquid hold-up volume and phase ratio (aqueous phase/organic phase, A/O) of the HNO3 –DtBuCH18C6/n-octanol system in a ϕ20 ACE were systematically investigated to find the cause of the above-mentioned hydrodynamic performance problem of the CESE process using the liquid-fast-separation method. It is shown that the operating and geometrical parameters have effects on the liquid hold-up volume and phase ratio (A/O). Moreover, the phase ratio (A/O) in the separation zone of the rotor is less than 1.5 under most of the present experimental conditions, which shows organic-phase entrainment may easily occur in the outlet aqueous phase. The cause can be that the density difference of two phases is small, viscosity of the organic phase is large, and interfacial tension of the extraction system is low.

A Predictive Modeling Approach to Support the Overfill Volume Definition of Liquid-in-Vial Drug Products.

Liquid-in-vial drug products are typically overfilled to meet the label claim volume specification while taking into account losses in the container-closure system and withdrawal device. Any overfill volume setting requires justification. The aim of this study was to estimate the overfill volume required for a liquid drug product in a vial using a prediction model. Glass vials sized from 2R to 20R capacity were filled with sorbitol-based aqueous solutions having a viscosity at 20°C ranging from 1 to 40 mPa·s. Viscosity and vial neck diameter were shown to be the main contributors to the hold-up volume of sorbitol-based aqueous solutions in vial and withdrawal syringe. The hold-up volume of various molecules of therapeutic interest was successfully estimated using a model built from sorbitol-based aqueous solutions data. A total variability approach is proposed for estimating the overfill volume of liquid-in-vial drug products, taking into account the product hold-up volume in vial and withdrawal syringe, the filling variability, and the extractable volume test variability. This prediction model could provide a first guess of the fill volume range to be tested to support overfill volume definition.

Evaluation of Hold-Up Volume Determination Methods and Markers in Hydrophilic Interaction Liquid Chromatography

Evaluation of Hold-Up Volume Determination Methods and Markers in Hydrophilic Interaction Liquid Chromatography

A Systematic Approach to Evaluating Closed System Drug-Transfer Devices During Drug Product Development

The use of Closed System Drug-Transfer Devices (CSTDs) has increased significantly in recent years due to NIOSH and USP recommendations to use them during preparation of hazardous drugs. Mechanistic and material differences between CSTDs and traditional in-use components warrant an assessment of their impact on product quality and dosing accuracy. Using a combination of prevalent CSTDs with biologic molecules, we performed an extensive assessment of the effect of using CSTDs for dose preparation and observed no negative impact on product quality attributes. Additionally, we found that the CSTD hold-up volume is 2 to 4-fold higher than conventional in-use components and exhibited a strong dependence on the CSTD brand used. We also found that the CSTD brand and dosing volume have a major influence on dosing accuracy with suboptimal protein recovery at very low dosing volumes. We identified entrapment of product in the CSTD spike as the root cause for this sub-optimal recovery and found that flushing the CSTD spike with a brand-new syringe and not the dosing syringe aided in complete protein recovery. Taken together we present a systematic approach to evaluate the risks and impact of CSTD to drug product quality, dose preparation, and dosing accuracy.

Considerations for Defining the Intravenous Administration Procedure for Nano-dose Sterile Drug Product in Clinical Studies

The loss of active substance, both small and large molecules, from sterile liquid drug products after contact with an administration kit has been extensively reported in the literature. This loss has been reported to be caused by incompatibility of the active substances with the contact surfaces of the administration kit and adsorption or sticking of the active substance to the surfaces of the administration kit. This paper investigates the mechanism for loss of a highly potent active substance based on the type and design of the administration kit. Two administration kits (syringe/Insyte Catheter and syringe/Nexiva Catheter) of different designs were used to administer a solution formulation of an ultra-low dose (nanograms) of a model hydrophobic active substance Compound X. The Nexiva Catheter was longer with tubing and Y connectors while the Insyte Catheter was shorter with no split septum tubing. Dose recovery from both administration kits was determined using high pressure liquid chromatography. The results indicated that the full dose was recovered from the syringes and Insyte Catheter. However, there was a significant loss of active substance from the Nexiva Catheter configuration even after post administration flush, which was due to holdup volume of the formulation within dead spaces of the Nexiva Catheter. It was also demonstrated that the dose recovery from the Nexiva Catheter can be significantly increased with increase in the post administration flush volume, which further confirms that the observed loss of active substance was not due to incompatibility or surface adsorption. The significance of this work is to provide awareness to formulation scientists that closed system Catheter design with Y connectors can be the main contributor for the loss in active substance, especially at ultra-low doses, and therefore dose recovery experiments should be expanded to include proper flushing of the Y connectors to expel any holdup volume from the Catheter.

Advanced score system and automated search strategies for parameter estimation in mechanistic chromatography modeling

Least squares estimation of unknown parameters from measurement data is a well-established standard method in chromatography modeling but can suffer from critical disadvantages. The description of real-world systems is generally prone to unaccounted mechanisms, such as dispersion in external holdup volumes, and systematic measurement errors, such as caused by pump delays. In this scenario, matching the shape between simulated and measured chromatograms has been found to be more important than the exact peak positions. We have therefore developed a new score system that separately accounts for the shape, position and height of individual peaks. A genetic algorithm is used for optimizing these multiple objectives. Even for non-conflicting objectives, this approach shows superior convergence in comparison to single-objective gradient search, while conflicting objectives indicate incomplete models or inconsistent data. In the latter case, Pareto optima provide important information for understanding the system and improving experiments. The proposed method is demonstrated with synthetic and experimental case studies of increasing complexity. All software is freely available as open source code (https://github.com/modsim/CADET-Match).

A New Definition of the Stationary Phase Volume in Mixed-Mode Chromatographic Columns in Hydrophilic Liquid Chromatography.

Polar columns used in the HILIC (Hydrophilic Interaction Liquid Chromatography) systems take up water from the mixed aqueous–organic mobile phases in excess of the water concentration in the bulk mobile phase. The adsorbed water forms a diffuse layer, which becomes a part of the HILIC stationary phase and plays dominant role in the retention of polar compounds. It is difficult to fix the exact boundary between the diffuse stationary and the bulk mobile phase, hence determining the column hold-up volume is subject to errors. Adopting a convention that presumes that the volume of the adsorbed water can be understood as the column stationary phase volume enables unambiguous determination of the volumes of the stationary and of the mobile phases in the column, which is necessary for obtaining thermodynamically correct chromatographic data in HILIC systems. The volume of the aqueous stationary phase, Vex, can be determined experimentally by frontal analysis combined with Karl Fischer titration method, yielding isotherms of water adsorbed on polar columns, which allow direct prediction of the effects of the composition of aqueous–organic mobile phase on the retention in HILIC systems, and more accurate determination of phase volumes in columns and consistent retention data for any mobile phase composition. The n phase volume ratios of 18 columns calculated according to the new phase convention strongly depend on the type of the polar column. Zwitterionic and TSK gel amide and amine columns show especially strong water adsorption.

Selecting carrier material for efficient biomethanation of industrial biogas-CO2 in a trickle-bed reactor

Biogas upgrading via biomethanation of renewable H2 and the CO2 in biogas poses a potential key technology in the decarbonized energy system. This study uses a methanogenic trickle-bed reactor for upgrading raw biogas from an industrial anaerobic digester to natural gas-grade biomethane through ex situ biomethanation. The carrier material comprises a critical design parameter for trickle-bed reactors by supporting the catalytic methanogenic biofilm and thereby the interfacial area for H2 mass transfer. The present study applied an abiotic mass transfer-based screening of six different carrier types to identify a candidate material that can facilitate efficient H2 mass transfer in a biomethanation trickle-bed reactor. The screening included gas-liquid mass transfer experiments in an air-water system in addition to characterizations of the carriers’ surface area, dynamic liquid hold-up volumes, and external porosities. The abiotic experiments showed that the gas-liquid mass transfer coefficient is highly influenced by material type and cannot be predicted solely from differences in the carriers’ surface area. The abiotic characterizations led to the selection of crushed clay particles, whose application in biomethanation-based upgrading of raw biogas from an industrial anaerobic digester was examined. The clay particles facilitated an apparent volumetric mass transfer coefficient for H2 of 10 min−1, enabling CH4 production rates up to 6.1 L · L−1 · d−1 and more than 99 % H2 conversion over 30 days of continuous operation, where after accumulation of metabolic water in the packed bed impaired H2 mass transfer. Apart from the adverse effect of water accumulation, there was no indication that the maximum H2 mass transfer rate was reached, demonstrating the selected carrier material’s industrial potential for biomethanation-based biogas upgrading.