Computational prediction of blend time in a large-scale viral inactivation process for monoclonal antibodies biomanufacturing.

Viral inactivation (VI) is a process widely used across the pharmaceutical industry to eliminate the cytotoxicity resulting from trace levels of viruses introduced by adventitious agents. This process requires adding Triton X-100, a non-ionic detergent solution, to the protein solution and allowing sufficient time for this agent to inactivate the viruses. Differences in process parameters associated with vessel designs, aeration rate, and many other physical attributes can introduce variability in the process, thus making predicting the required blending time to achieve the desired homogeneity of Triton X-100 more critical and complex. In this study we utilized a CFD model based on the lattice Boltzmann method (LBM) to predict the blend time to homogenize a Triton X-100 solution added during a typical full-scale commercial VI process in a vessel equipped with an HE-3-impeller for different modalities of the Triton X-100 addition (batch vs. continuous). Although direct experimental progress of the blending process was not possible because of GMP restrictions, the degree of homogeneity measured at the end of the process confirmed that Triton X-100 was appropriately dispersed, as required, and as computationally predicted here. The results obtained in this study were used to support actual production at the biomanufacturing site.

Characterization of Turbulent Properties in the EPA Baffled Flask for Dispersion Effectiveness Testing.

The baffled flask test (BFT) has been proposed by United States Environmental Protection Agency to be adopted as the official standard protocol for testing dispersant effectiveness. The mixing energy in the baffled flask is investigated in this paper. Particle image velocimetry (PIV) was used to measure the water velocity in the flask placed at an orbital shaker that was rotated at seven rotation speeds: 100, 125, 150, 160, 170, 200, and 250 rpm. Two dimensional velocity fields in large and small vertical cross sections of the flask for each rotation speed were obtained. The one-dimensional (1D) energy spectra indicates the existence of inertial subrange. The estimated average energy dissipation rates were in the range 7.65×10-3 to 4 W/kg for rotation speeds of Ω=100-250 rpm, of which it is larger than the one estimated by prior studies using single-point velocity measurement techniques for Ω=100 and 200 rpm. Factors such as instruments used, velocity components measured, and different analysis methods could contribute to the discrepancies in the results. The Kolmogorov scale estimated in this study for all seven rotation speeds approached the size of oil droplets observed at sea, which is 50-400 µm. The average energy dissipation rate, ε and Kolmogorov microscale, η, in the flasks were correlated to the rotation speed, and it was found that ε ¯ = 9.0 × 10 - 5 Exp (0.043Ω) with R 2 = 0.97 and η ¯ = 1 , 463 Exp (-0.015Ω) with R 2 = 0.98.

Effect of basket mesh size on the hydrodynamics of a partially filled (500 mL) USP rotating basket dissolution testing Apparatus 1.

The USP Rotating Basket Dissolution Testing Apparatus 1 is listed in the USP as one of the tools to assess dissolution of oral solid dosage forms. Baskets of different mesh sizes can be used to differentiate between dissolution profiles of different formulations. Here, we used Particle Image Velocimetry (PIV) to study the hydrodynamics of the USP Apparatus 1 using baskets with different mesh openings (10-, 20- and 40-mesh) revolving at 100 rpm, when the vessel was filled with 500 mL. The velocity profiles throughout the liquid were found to vary significantly using baskets of different mesh sizes, typically increasing with increased size of the opening of the basket mesh, especially for axial and radial velocities. This, in turn, resulted in a significantly different flow rate through the basket, which can be expected to significantly impact the dissolution rate of the drug product. A comparison between the results of this work with those of a previous study with a 900-mL fill volume (Sirasitthichoke et al., Intern. J. Pharmaceutics, 2021, 607: 120976), shows that although the hydrodynamics in the USP Apparatus 1 changed with fill level in the vessel, the flow rate through the basket was not significantly affected. This implies that tablets dissolving in the two systems would experience similar tablet-liquid medium mass transfer coefficients, and therefore similar initial dissolution rates, but different dissolution profiles because of the difference in volume.

Particle Image Velocimetry (PIV) measurements of USP Apparatus 1 hydrodynamics with 500 mL fill volume.

Changes to hydrodynamics arising from changes within dissolution testing systems, such as the fill volume level, can potentially cause variability in dissolution results. However, the literature on hydrodynamics in Apparatus 1 is quite limited and little information is available for vessels with different liquid volumes. Here, velocities in a USP Apparatus 1 vessel with a liquid fill volume of 500 mL, a common alternative to 900 mL, were experimentally measured using 2D-2C Particle Image Velocimetry (PIV) for different basket rotational speeds. Tangential velocities dominated the flow field, while axial and radial velocities were much lower and varied with location. The velocities distribution increased proportionately with the basket rotational speed almost everywhere in the vessel excepting for underneath the basket. A nearly horizontal radial liquid jet was found to originate close to the basket upper edge. Comparison of these results with those previously reported with 900-mL liquid volume (Sirasitthichoke et al., Intern. J. Pharmaceutics:X; 3 (2021) 100078) showed that the flow rate through the baskets was similar in both systems, implying that, at least initially, the amount of drug in solution would increase linearly with time. In other words, the flow rate through the baskets would be independent of the liquid volume. Velocity profiles were also found to be similar, except in the region above the basket, which was affected by the radial jet with an orientation significantly different between the 500-mL and the 900-mL systems.

Experimental determination of the velocity distribution in USP Apparatus 1 (basket apparatus) using Particle Image Velocimetry (PIV).

The USP Apparatus 1 (basket apparatus) is commonly used to evaluate the dissolution performance of oral solid dosage forms. The hydrodynamics generated by the basket contributes, in general, to the dissolution rate and hence the dissolution results. Here, the hydrodynamics of Apparatus 1 was quantified in a vessel filled with 900-mL de-ionized water at room temperature by determining, via Particle Image Velocimetry (PIV), the velocity profiles on a vertical central plane and on 11 horizontal planes at different elevations at three different basket agitation speeds. The flow field was dominated by the tangential velocity component and was approximately symmetrical in all cases. Despite all precautions taken, small flow asymmetries were observed in the axial and radial directions. This appears to be an unavoidable characteristic of the flow in Apparatus 1. The magnitudes of the axial and radial velocity components varied with location but were always low. A small jet was seen emanating radially near the top edge of the basket. Velocities typically scaled well with increasing agitation speed in most regions of the vessel except for a region directly below the basket. The results of this work provide a major insight into the flow field inside the USP Apparatus 1.

Influence of basket mesh size on the hydrodynamics in the USP rotating basket dissolution testing Apparatus 1.

The USP Apparatus 1 (rotating basket), typically used to assess drug product reproducibility and evaluate oral solid dosage forms performance, consists of a cylindrical glass vessel with a hemispherical bottom and a wire basket rotating at constant speed. Baskets with different wire openings can be used in alternative to the standard mesh opening (40-mesh) in order to discriminate between drug formulations during early stage of drug product development. Any changes introduced by different basket geometries can potentially and significantly impact the system hydrodynamics and cause variability of results, thus affecting product quality. In this work, Particle Image Velocimetry (PIV) was used to experimentally quantify the velocity distribution in the USP rotating basket Apparatus 1 using baskets of different mesh sizes (10-, 20-, and 40-mesh size) under the typical operating conditions described in dissolution testing procedures. Similar flow patterns were observed in all cases. However, the radial and axial velocities in the USP Apparatus 1 generally increased with increasingly larger openings of the basket mesh. Increasing the basket agitation speed also resulted in an overall increase in the velocities, especially below in the innermost core region below the basket, where drug fragments typically reside. More importantly, the flow entering and leaving the baskets was quantified from the velocity profiles in the immediate vicinity of the baskets. It was found that the flow increased significantly with increasingly larger mesh openings, which can, in turn, promote faster dissolution of the oral solid dosage forms, thus affecting drug dissolution profiles. Hence, the selection of the basket mesh size must be carefully considered during drug product development.

Cetylpyridinium Trichlorostannate: Synthesis, Antimicrobial Properties, and Controlled-Release Properties via Electrical Resistance Tomography.

Cetylpyridinium trichlorostannate (CPC-Sn), comprising cetylpyridinium chloride (CPC) and stannous chloride, was synthesized and characterized via single-crystal X-ray diffraction measurements indicating stoichiometry of C21H38NSnCl3 where the molecules are arranged in a 1:1 ratio with a cetylpyridinium cation and a [SnCl3]- anion. CPC-Sn has shown potential for application as a broad-spectrum antimicrobial agent, to reduce bacteria-generated volatile sulfur compounds and to produce advanced functional materials. In order to investigate its controlled-release properties, electrical resistance tomography was implemented. The results demonstrate that CPC-Sn exhibits extended-release properties in an aqueous environment as opposed to the CPC counterpart.

Computational hydrodynamic comparison of a mini vessel and a USP 2 dissolution testing system to predict the dynamic operating conditions for similarity of dissolution performance.

The hydrodynamic characteristics of a mini vessel and a USP 2 dissolution testing system were obtained and compared to predict the tablet-liquid mass transfer coefficient from velocity distributions near the tablet and establish the dynamic operating conditions under which dissolution in mini vessels could be conducted to generate concentration profiles similar to those in the USP 2. Velocity profiles were obtained experimentally using Particle Image Velocimetry (PIV). Computational Fluid Dynamics (CFD) was used to predict the velocity distribution and strain rate around a model tablet. A CFD-based mass transfer model was also developed. When plotted against strain rate, the predicted tablet-liquid mass transfer coefficient was found to be independent of the system where it was obtained, implying that a tablet would dissolve at the same rate in both systems provided that the concentration gradient between the tablet surface and the bulk is the same, the tablet surface area per unit liquid volume is identical, and the two systems are operated at the appropriate agitation speeds specified in this work. The results of this work will help dissolution scientists operate mini vessels so as to predict the dissolution profiles in the USP 2, especially during the early stages of drug development.

Experimental and computational determination of blend time in USP Dissolution Testing Apparatus II.

Blend time, the time to achieve a predefined level of homogeneity of a tracer in a mixing vessel, is an important parameter to evaluate the mixing efficiency of mixing devices. In this work, the blend time required to homogenize the liquid content of a USP Dissolution Testing Apparatus II under a number of operating conditions was obtained using two different experimental methods (tracer detection via colorimetric and conductivity measurements), a computational approach (computational fluid dynamics (CFD)), and a semi-theoretical analysis of the phenomenon. Under the standard geometric and operating conditions in which the USP Apparatus II is typically used (N = 50 rpm) the experimental blend time to achieve a 92.74% uniformity level was found to be between 27.5 and 33.3 s, depending on the location of the injection point and monitoring point for the tracer. These values were in close agreement with those obtained from CFD simulations. Changing the impeller vertical position (+/-2 mm) had only a limited effect. The CFD predictions also indicated that blend time is inversely proportional to the agitation speed. This conclusion is in agreement with previous reports and equations for blend time in mixing vessels. The blend times obtained in this work appear to be some two orders of magnitude smaller than the time usually required for appreciable tablet dissolution during the typical dissolution test, implying that the liquid contents of the USP Apparatus II can be considered to be relatively well mixed during the typical dissolution test.

Hydrodynamic investigation of USP dissolution test apparatus II.

The USP Apparatus II is the device commonly used to conduct dissolution testing in the pharmaceutical industry. Despite its widespread use, dissolution testing remains susceptible to significant error and test failures, and limited information is available on the hydrodynamics of this apparatus. In this work, laser-Doppler velocimetry (LDV) and computational fluid dynamics (CFD) were used, respectively, to experimentally map and computationally predict the velocity distribution inside a standard USP Apparatus II under the typical operating conditions mandated by the dissolution test procedure. The flow in the apparatus is strongly dominated by the tangential component of the velocity. Secondary flows consist of an upper and lower recirculation loop in the vertical plane, above and below the impeller, respectively. A low recirculation zone was observed in the lower part of the hemispherical vessel bottom where the tablet dissolution process takes place. The radial and axial velocities in the region just below the impeller were found to be very small. This is the most critical region of the apparatus since the dissolving tablet will likely be at this location during the dissolution test. The velocities in this region change significantly over short distances along the vessel bottom. This implies that small variations in the location of the tablet on the vessel bottom caused by the randomness of the tablet descent through the liquid are likely to result in significantly different velocities and velocity gradients near the tablet. This is likely to introduce variability in the test.

Reduction of 2,4-dichlorophenol toxicity to Pseudomonas putida after oxidative incubation with humic substances and a biomimetic catalyst.

The effect of a synthetic iron(III)-porphyrin meso-tetra(2,6-dichloro-3-sulfonatophenyl)porphyrinate as a biomimetic catalyst in the oxidative treatment of 2,4-dichlorophenol (2,4-DCP) with humic substances and H(2)O(2) was evaluated in factorial design experiments conducted at different concentrations of 2,4-DCP (0-25 ppm) and different incubation treatment times (0, 24, 96, or 120 h). In the absence of this treatment, bioassays with the bacterium Pseudomonas putida (ATCC11250) showed decreasing specific growth rates mu (used here to quantify 2,4-DCP toxicity) with increasing concentrations of 2,4-DCP. However, when 2,4-DCP was treated as mentioned above the toxicity of the resulting 2,4-DCP solution was reduced significantly. At low 2,4-DCP concentrations (up to 5 ppm) and long incubation periods (as long as 120 h), the specific growth rate mu was comparable to that of cultures grown in the absence of 2,4-DCP. The reduction in toxicity was directly correlated to a decrease in the concentration of 2,4-DCP in the treated solutions, as measured by high-performance liquid chromatography. The reduced concentrations of 2,4-DCP in the treated solutions could be correctly predicted based on the relationship between the specific growth rates and the 2,4-DCP concentrations in untreated solutions. These results indicate that the oxidative coupling of 2,4-DCP to humic substances catalyzed by the synthetic iron(III)-porphyrin catalyst in the presence of H(2)O(2) is responsible for the removal of 2,4-DCP from solutions. This approach appears to be a promising alternative treatment to reduce 2,4-DCP bioavailability and thus toxicity in the environment.

Experimental and computational determination of the hydrodynamics of mini vessel dissolution testing systems.

Mini vessel dissolution testing systems consist of a small-scale 100-mL vessel with a small paddle impeller, similar to the USP Apparatus 2, and are typically utilized when only small amounts of drug product are available during drug development. Despite their common industrial use, mini vessels have received little attention in the literature. Here, Computational Fluid Dynamics (CFD) was used to predict velocity profiles, flow patterns, and strain rate distribution in a mini vessel at different agitation speeds. These results were compared with experimental velocity measurements obtained with Particle Image Velocimetry (PIV). Substantial agreement was observed between CFD results and PIV data. The flow is strongly dominated by the tangential velocity component. Secondary flows consist of vertical upper and lower recirculation loops above and below the impeller. A low recirculation zone was observed in the lower part of the vessel. The radial and axial velocities in the region just below the impeller are very small especially in the innermost core zone below the paddle, where tablet dissolution occurs. Increasing agitation speed reduces the radius of this zone, which is always present at any speed, and only modestly increases the tangential flow intensity, with significant implication for dissolution testing in mini vessels.

Dissolution of prednisone tablets in the presence of an arch-shaped fiber optic probe in a USP dissolution testing apparatus 2.

During dissolution testing of solid dosage forms in the United States Pharmacopoeia (USP) Apparatus 2, samples are manually withdrawn from the medium in the vessel prior to the analysis. Probes permanently inserted in the medium can automate the sampling process but can also alter the system's hydrodynamics, possibly resulting in different dissolution-testing results. In this work, dissolution tests were conducted in a USP Apparatus 2 with and without an arch-shaped fiber optic probe using prednisone tablets fixed at nine different locations on the vessel bottom. The resulting dissolution profiles were compared using statistical tools. Dissolution rates obtained with the probe were typically higher than those obtained without the probe. The magnitude of the difference between dissolution profiles depended on the tablet location: Larger differences were observed with tablets located immediately downstream of the probe. The differences in dissolution profiles were generally small enough to satisfy the US Food and Drug Administration criteria (f1 and f2 values), although a paired t-test [P(t-test)] indicated that most of the profiles were statistically different [P(t-test) <0.05]. The hydrodynamic effects generated by the arch-shaped fiber optic probe resulted in detectable differences in the dissolution profiles, which, although limited, were clearly measurable and could introduce variations in test results.

A novel off-center paddle impeller (OPI) dissolution testing system for reproducible dissolution testing of solid dosage forms.

Dissolution testing is routinely conducted in the pharmaceutical industry to provide in vitro drug release information for quality control purposes. The most common dissolution testing system for solid dosage forms is the United States Pharmacopeia (USP) Dissolution Testing Apparatus 2. This apparatus is very sensitive to the initial location of the tablet, which cannot be controlled because the tablet is dropped into the vessel at the beginning of the test and it may rest at random locations at the vessel's bottom. In this work, a modified Apparatus 2 in which the impeller was placed 8 mm off center in the vessel was designed and tested. This new design was termed "OPI" for "off-center paddle impeller." Dissolution tests were conducted with the OPI apparatus for nine different tablet locations using both disintegrating tablets (prednisone) and nondisintegrating tablets (salicylic acid). The dissolution profiles in the OPI apparatus were largely independent of the tablet location at the vessel's bottom, whereas those obtained in the Standard System generated statistically different profiles depending on the tablet location. The newly proposed OPI system can effectively eliminate artifacts generated by random settling of the tablet at the vessel's bottom, thus making the test more robust.

Velocity profiles and shear strain rate variability in the USP Dissolution Testing Apparatus 2 at different impeller agitation speeds.

The fluid velocity profiles at different locations inside a standard USP Dissolution Testing Apparatus 2 were experimentally obtained via Laser Doppler Velocimetry (LDV) at three impeller agitations speeds, namely 50rpm, 75rpm and 100rpm. The experimental results were compared with the predictions obtained with Computational Fluid Dynamics (CFD) where the κ-ω model with low Reynolds number correction was used to account for turbulence effects. In general, good agreement was found between the experimental LDV velocity measurements and the CFD simulation predictions. The non-dimensional tangential, axial and radial velocity profiles (scaled with the impeller tip speed) and the flow pattern were found to be nearly independent of the agitation speed in most regions of the vessel, implying that increasing the agitation speed generally produced a corresponding increase in the local values of the velocity. However, the velocity profiles and flow pattern in the inner core region just below the impeller, where the dissolving tablet is usually located, were found to be much less sensitive to agitation speed. In this region, the axial and radial velocities were especially low and were not significantly affected by agitation increases. This inner core region at the center of the vessel bottom persisted irrespective of agitation intensity. The CFD predictions also indicated that increasing the agitation speed resulted in a higher shear strain rate distribution along the vessel bottom, although the strain rate was always very low at the center of the vessel bottom, even when the agitation speed was increased.

Hydrodynamic, mass transfer, and dissolution effects induced by tablet location during dissolution testing.

Tablets undergoing dissolution in the USP Dissolution Testing Apparatus II are often found at locations on the vessel bottom that are off-center with respect to the dissolution vessel and impeller. A previously validated CFD approach and a novel experimental method were used here to examine the effect of tablet location on strain rates and dissolution rates. Dissolution tests were conducted with non-disintegrating tablets (salicylic acid) and disintegrating tablets (Prednisone) immobilized at different locations along the vessel bottom. CFD was used to predict the velocity profiles and strain rates when the tablets were placed at such locations. A CFD-based model was derived to predict the mass transfer coefficient and dissolution curves, which were then compared to the experimental results. Both non-disintegrating and disintegrating off-center tablets experimentally produced higher dissolution rates than centered tablets. The CFD-predicted strain rate distribution along the bottom was highly not uniform and the predicted strain rates correlated well with the experimental mass transfer coefficients. The proposed CFD-based model predicts mass transfer rates that correlate well with the experimental ones. The exact tablet location has a significant impact on the dissolution profile. The proposed model can satisfactorily predict the mass transfer coefficients and dissolution profiles for non-disintegrating tablets.

Velocity distribution and shear rate variability resulting from changes in the impeller location in the USP dissolution testing apparatus II.

PURPOSE: The United States Pharmacopoeia (USP) imposes strict requirements on the geometry and operating conditions of the USP Dissolution Testing Apparatus II. A previously validated Computational Fluid Dynamics (CFD) approach was used here to study the hydrodynamics of USP Apparatus II when the impeller was placed at four different locations, all within the limits specified by USP. METHOD: CFD was used to predict the velocity profiles, energy dissipation rates, and strain rates when the impeller was placed in the reference location (centrally mounted, 25 mm off the vessel bottom), 2 mm off-center, 2 mm higher, and 2 mm lower than the reference location. RESULTS: Small changes in impeller location, especially if associated with loss of symmetry, produced extensive changes in velocity profiles and shear rates. Centrally located impellers, irrespective of their off-bottom clearance, produced non-uniform but nearly symmetric strain rates. The off-center impeller produced a more uniform but slightly asymmetric strain rate distribution. CONCLUSIONS: The system hydrodynamics depends strongly on small differences in equipment configurations and operating conditions, which are likely to affect significantly the flow field and shear rate experienced by the oral dosage form being tested, and hence the solid-liquid mass transfer and dissolution rate.