Calcium oxalate (CaOx) crystallization under laminar flow conditions, relevant for kidney stone formation, was studied in a microfluidic device simulating the geometry of kidney collecting ducts. In a typical microfluidic experiment, two reactive solutions with designated concentrations of calcium (Ca) and oxalate (Ox) ions were brought into contact in a microfluidic channel to create a laminar co-current flow of the two streams. As the streams flow co-currently in the channel, diffusion takes place between the two streams across the channel width, resulting in reactive crystallization leading to CaOx nucleation and growth of CaOx crystals along the mixing front. We studied the growth of these crystals in artificial urine as a function of the fluid flow rate in the channel, the molar ratio of Ca : Ox in the medium and the presence of an organic protein, osteopontin (OPN), known to inhibit the growth of CaOx crystals. Three different flow velocities at a fixed molar ratio of Ca : Ox = 7.5 and four molar ratios of Ca : Ox at a fixed mean flow velocity of 0.035 m s−1 were tested. Lastly, three additive OPN concentrations were evaluated: 2.4 × 10−8 mol m−3, 6 × 10−8 mol m−3 and 8.4 × 10−8 mol m−3. The mean flow velocity did not alter the crystal growth of CaOx in the studied range, whereas altering the molar ratio of Ca : Ox had a high impact on the growth rate. In addition, the type of pseudopolymorph which nucleated appears to depend strongly on the molar ratio. At a low Ca : Ox ratio, both calcium oxalate monohydrate (COM) and calcium oxalate dihydrate (COD) nucleated simultaneously and the growth of the two pseudopolymorphic forms of CaOx crystals was observed. The lowest applied OPN concentration decreased the growth rate of COD, while higher concentrations of OPN slowed down the nucleation kinetics to a point that it completely inhibited the formation of any CaOx crystal in artificial urine within the investigated timeframe. COD was seen under all the conditions investigated, whilst COM was seen in experiments for Ca : Ox molar ratio values between 5 and 6. Our results were rationalized using finite element simulations supported by solution chemistry modelling.@en

Non-photochemical laser-induced nucleation (NPLIN) has emerged as a promising primary nucleation control technique offering spatiotemporal control over crystallization with potential for polymorph control. So far, NPLIN was mostly investigated in milliliter vials, through laborious manual counting of the crystallized vials by visual inspection. Microfluidics represents an alternative to acquiring automated and statistically reliable data. Thus we designed a droplet-based microfluidic platform capable of identifying the droplets with crystals emerging upon Nd:YAG laser irradiation using the deep learning method. In our experiments, we used supersaturated solutions of KCl in water, and the effect of laser intensity, wavelength (1064, 532, and 355 nm), solution supersaturation (S), solution filtration, and intentional doping with nanoparticles on the nucleation probability is quantified and compared to control cooling crystallization experiments. Ability of dielectric polarization and the nanoparticle heating mechanisms proposed for NPLIN to explain the acquired results is tested. Solutions with lower supersaturation (S = 1.05) exhibit significantly higher NPLIN probabilities than those in the control experiments for all laser wavelengths above a threshold intensity (50 MW/cm²). At higher supersaturation studied (S = 1.10), irradiation was already effective at lower laser intensities (10 MW/cm²). No significant wavelength effect was observed besides irradiation with 355 nm light at higher laser intensities (≥50 MW/cm²). Solution filtration and intentional doping experiments showed that nanoimpurities might play a significant role in explaining NPLIN phenomena.

@en

Herein, we study the influences of the laser-exposed volume and the irradiation position on the nonphotochemical laser-induced nucleation (NPLIN) of supersaturated potassium chloride solutions in water. The effect of the exposed volume on the NPLIN probability was studied by exposing distinct milliliter-scale volumes of aqueous potassium chloride solutions stored in vials at two different supersaturations (1.034 and 1.050) and laser intensities (10 and 23 MW/cm²). Higher NPLIN probabilities were observed with increasing laser-exposed volume as well as with increasing supersaturation and laser intensity. The measured NPLIN probabilities at different exposed volumes are questioned in the context of the dielectric polarization mechanism and classical nucleation theory. No significant change in the NPLIN probability was observed when samples were irradiated at the bottom, top, or middle of the vial. However, a significant increase in the nucleation probability was observed upon irradiation through the solution meniscus. We discuss these results in terms of mechanisms proposed for NPLIN.

@en

The increasing prevalence of urolithiasis in industrialized societies triggered considerable interest in how various species found in urine regulate the nucleation and growth of common kidney stone constituents such as calcium oxalate (CaOx). Yet, the role macromolecules play in kidney stone formation is often overlooked due to their low concentration in urine. In this study, we investigate the nucleation kinetics of CaOx in artificial urine with droplet-based microfluidic induction time measurements at varying concentrations of oxalate and hyaluronic acid (HA), a polysaccharide commonly found in urine. The formation of two pseudo-polymorphic forms of calcium oxalate crystals, calcium oxalate monohydrate (COM) and calcium oxalate dihydrate (COD), are carefully monitored using polarized light microscopy in induction time experiments. COM and COD nucleated concomitantly in artificial urine yet with distinct kinetics. Our results indicate that higher oxalate concentrations favor the formation of COD, the metastable form, over COM, the most stable form. Moreover, COD is also the fastest nucleating form in droplets under studied conditions. Furthermore, increasing the concentration of HA at fixed calcium and oxalate concentrations favored the nucleation of COM. We observed that in droplets where COM nucleated first, COD was not formed within the experimental time scale. However, in the droplets where COD appeared first, COM crystals were also observed later. We hope our findings shed light on the role macromolecules such as HA plays in dictating the pseudo-polymorphic form of CaOx and guide next generation treatments.@en

We present a droplet microfluidic platform mixing the contents of the droplet chaotically in microfluidic induction time measurements, a promising method for quantifying nucleation kinetics with minute amounts of solute. The nucleation kinetics of aqueous potassium chloride droplets dispersed in mineral oil without surfactants is quantified in the presence and absence of chaotic mixing. We demonstrate the ability of the proposed platform to dictate droplet size, to provide a homogeneous temperature distribution, and to chaotically mix the droplet contents. Chaotic mixing in induction time measurements is facilitated by the motion of droplets through serpentine micromixer bends, while the extent of mixing is controlled by how much droplets move. Different nucleation kinetics are observed in experiments where the droplets are static, mixed, and in motion. We hypothesize that the droplet motion induces formation of a thin-liquid Bretherton film surrounding the droplets. The thin film shields droplets from solid boundaries that are more efficient heteronucleant surfaces compared to liquid-liquid interfaces. We observed that repeated microfluidic induction time measurements, particularly with moving droplets, produce significantly distinct cumulative nucleation probability curves, indicating that the measured nucleation kinetics depend strongly on the details of the experimental procedure, which we discuss in detail. Finally, we compare the microfluidic experiments to well-mixed, milliliter volume, turbidity-based measurements in the context of classic nucleation theory.

@en

Nonphotochemical laser-induced nucleation (NPLIN) is a promising primary nucleation control method, yet its underlying mechanism remains elusive. To contribute to the discussion on whether the polarization of laser irradiation in NPLIN experiments influences the polymorphic outcome, we revisit NPLIN experiments with aqueous glycine solutions with supersaturations ranging between S = 1.5 and S = 1.7 irradiated by nanosecond pulses (∼7 ns) of near-infrared wavelength (1064 nm). Systematically altering laser light excitation properties, including the number of pulses and type of polarization, we quantified the nucleation kinetics and characterized the polymorphic form that crystallized upon laser irradiation. Due to the stochasticity of the nucleation process, a large number of samples (>100 per each experimental point) were studied under carefully controlled experimental conditions such as the ambient temperature, cooling rate, and aging period. We observed significant differences among laser-irradiated, spontaneously nucleated, and crash-cooled samples in terms of nucleation kinetics and polymorphic form. This result indicates that laser irradiation provides a different polymorph-forming pathway in comparison to crash-cooling and spontaneous nucleation. However, no clear dependence between the polymorphic form and the polarization of laser irradiation is observed. We discuss our results in the context of previous reports supported thorough quantification of sample heating in NPLIN experiments.

@en

Cocrystallization of racemic-compound-forming chiral molecules can result in conglomerate cocrystals or diastereomerically related cocrystals, which enable the application of chiral separation techniques such as preferential crystallization and classic resolution. Here, a systematic method to identify the types and phase diagrams of cocrystals formed by chiral target compounds and candidate coformers in a particular solvent system is presented, which allows the design of suitable chiral resolution processes. The method is based on saturation temperature measurements of specific solution compositions containing both enantiomers of chiral molecules and a coformer. This method is applied to analyze three different systems. For racemic phenylalanine (Phe) in water/ethanol mixtures one of the enantiomers selectively cocrystallizes with the opposite enantiomer of valine (Val), forming the more stable diastereomerically related cocrystal. The racemic compound ibuprofen crystallizes with the nonchiral coformer 1,2-bis(4-pyridyl)ethane (BPN) as racemic compound cocrystals. More interestingly, when it is combined with trans-1-(2-pyridyl)-2-(4-pyridyl)ethylene (BPE), the racemic compound ibuprofen cocrystallizes as a conglomerate, which in principle enables the application of preferential crystallization of this racemic compound. The systematic method shows the benefit of using pseudo-binary phase diagrams. Such pseudo-binary phase diagrams depict the saturation temperature on a very specific route through the quaternary phase diagram, allowing the identification of various cocrystal types as well as the corresponding cocrystallization conditions. The systematic method can be used to identify a suitable solid phase for chiral separation, and the obtained phase diagram information enables the performance of a crystallization-mediated chiral resolution process design. Such a guideline for a chiral resolution process design has never been reported for conglomerate cocrystal systems such as IBU:BPE, presented in this study.

@en

A droplet-based microfluidic platform is presented to study the nucleation kinetics of calcium oxalate monohydrate (COM), the most common constituent of kidney stones, while carefully monitoring the pseudo-polymorphic transitions. The precipitation kinetics of COM is studied as a function of supersaturation and pH as well as in the presence of inhibitors of stone formation, magnesium ions (Mg2+), and osteopontin (OPN). We rationalize the trends observed in the measured nucleation rates leveraging a solution chemistry model validated using isothermal solubility measurements. In equimolar calcium and oxalate ion concentrations with different buffer solutions, dramatically slower kinetics is observed at pH 6.0 compared to pHs 3.6 and 8.6. The addition of both Mg2+ and OPN to the solution slows down kinetics appreciably. Interestingly, complete nucleation inhibition is observed at significantly lower OPN, namely, 3.2 × 10-8 M, than Mg2+ concentrations, 0.875 × 10-4 M. The observed inhibition effect of OPN emphasizes the often-overlooked role of macromolecules on COM nucleation due to their low concentration presence in urine. Moreover, analysis of growth rates calculated from observed lag times suggests that inhibition in the presence of Mg2+ cannot be explained solely on altered supersaturation. The presented study highlights the potential of microfluidics in overcoming a major challenge in nephrolithiasis research, the overwhelming physiochemical complexity of urine.

@en

Accurate Calcium Oxalate Monohydrate (COM) solubility measurements are essential for elucidating the physiochemical mechanism behind the formation of kidney stones, nephrolithiasis. Yet the reported solubility values of COM in ultrapure water, arguably the simplest solvent relevant for nephrolithiasis, vary significantly depending on implemented method. To address this variation, we present an experimental study of the solubility of COM validated by a model based on the Debye–Hückel theory describing the solution chemistry and the complex formation. We also carefully monitor potential pseudopolymorphic/hydrate transitions during the solubility measurements with in-situ and ex-situ methods. Our results indicate that the solubility of COM in ultrapure water is a weak function of temperature. However, the measured solubility varies significantly in buffer solutions across physiologically relevant pH values at body temperature. The proposed model explains observed trends as a combined effect of ionic strength, protonation reactions, and soluble complex formation. Moreover, it predicts solubility of COM in buffer solutions remarkably well using our measurements in ultrapure water as input, demonstrating the consistency of presented approach. The presented study parleying experiments and modelling provides a solid stepping stone to extend the physiochemical understanding of nephrolithiasis to more realistic solutions laden with biological complexity.@en

In this study, an in situ imaging system has been analysed to characterize the crystal size, the shape and the number of particles during a continuous crystallization process in a Continuous Oscillatory Baffled Crystallizer (COBC). Two image analysis approaches were examined for particle characterization in the suspension containing both small nuclei and larger grown crystals (nonspherical and irregular in shape). The pattern matching approach, in which the particles are approximated to be spherical, did result in an overestimation of the size. Alternatively, a segmentation-based algorithm resulted in reliable crystal size and shape characteristics. The laser diffraction analysis in comparison to the image analysis overestimated the particle sizes due to the agglomeration of particles upon filtration and drying. The trend in the particle counts during the start of crystallization process, including nucleation, determined by the image analysis probe was comparable with the one measured by FBRM, highlighting the potential of in situ imaging for process monitoring.

@en

Various mechanisms have been proposed to explain the nonphotochemical laser-induced nucleation (NPLIN). Identifying the dominant mechanism requires addressing a large set of experimental parameters with a statistically significant number of samples, forced by the stochastic nature of nucleation. In this study, with aqueous KCl system, we focus on the nucleation probability as a function of laser wavelength, laser intensity, and sample supersaturation, whereas the influence of filtration and the laser-induced radiation pressure on NPLIN activity is also studied. To account for the nucleation stochasticity, we used 80-100 samples. The NPLIN probability showed an increase with increasing laser intensity. The results are different from the previous report, as a supersaturation independent intensity threshold is not observed. No dependence of the NPLIN probability on the laser wavelength (355, 532, and 1064 nm) was observed. Filtration of samples reduced the nucleation probability suggesting a pronounced role of impurities on NPLIN. The magnitude and the propagation velocity of the laser-induced radiation pressure were quantified using a pressure sensor under laser intensities ranging from 0.5 to 80 MW/cm². No correlation was found between the radiation pressure and NPLIN at our unfocused laser beam intensities ruling out the radiation pressure as a possible cause for nucleation.

@en

In this research, a process model has been developed for an airlift and compared with that of a draft tube stirred crystallizer to clarify the crystallization kinetics in this novel type of crystallizer. Recently it has been shown that although secondary nucleation is strongly suppressed in this crystallizer, it is not completely absent and further development and scale up requires a more quantitative prediction of the kinetics in this type of crystallizer. A number of growth and nucleation models were examined in a parameter estimation study in which a dataset of a number of seeded batch experiments has been used to estimate the kinetics in both an airlift and a draft tube stirred crystallizer. It is shown that a kinetic model, consisting of a two-step growth model and two additive nucleation mechanisms, i.e. an attrition based and a surface nucleation mechanism, gives an excellent and statistically acceptable description of all studied experiments with one parameter set each for both type of crystallizers. The main difference in the two types of crystallizers being that the attrition terms by crystal-impeller and crystal–crystal collisions in the airlift crystallizer can be completely neglected.@en

The use of a continuously operated oscillatory flow baffled crystallizer (COBC) has been promoted as a promising alternative crystallizer design for continuous crystallization because of the claim, based on dispersion of liquid, that plug flow can be achieved. Plug flow can lead to uniformity in product quality, if good control over nucleation and the growth of crystals is also achieved. In this study a residence time distribution (RTD) analysis was made for both homogeneous (methylene blue-water) and heterogeneous tracer system (melamine-water). In literature it is proposed, on the basis of homogeneous tracer experiments only, that the velocity ratio ΨΨ (the ratio between the oscillatory velocity and the superficial velocity of the imposed flow) is sufficient to identify optimal operating conditions for plug flow in COBC. Multiple combinations of amplitude and frequency result in the desired ΨΨ value. Our results show that operating at high amplitudes increases dispersion, reducing the plug flow like mixing. Thus ΨΨ alone is not sufficient for optimizing the mixing. Our study for the first time compares dispersion of homogenous and heterogeneous tracer in the commercially available DN15 system, addressing the knowledge gap in handling solids in COBC. Comparable responses were obtained with both the tracer systems for changes in the oscillatory flow variables. Operation at relative low amplitudes was optimal to obtain plug flow like behavior, even with 10% (w/w) slurry with no problems of the particles settling. The optimal operating condition for minimal dispersion was clearly different for the homogenous and the heterogeneous system.@en

The control of nucleation in crystallization processes is a challenging task due to the often lacking knowledge on the process kinetics. Inflexible (predetermined) control strategies fail to grow the nucleated crystals to the desired quality because of the variability in the process conditions, disturbances, and the stochastic nature of crystal nucleation. Previously, the concept of microwave assisted direct nucleation control (DNC) was demonstrated in a laboratory setup to control the crystal size distribution in a batch crystallization process by manipulating the number of particles in the system. Rapid temperature cycling was used to manipulate the super(under)saturation and hence the number of crystals. The rapid heating response achieved with the microwave heating improved the DNC control efficiency, resulting in halving of the batch time. As an extension, this work presents a novel design in which the microwave applicator is integrated in the crystallizer, hence avoiding the external loop though the microwaves oven. DNC implemented in the 4 L unseeded crystallizer, at various count set points, resulted in strong efficiency enhancement of DNC, when compared to the performance with a slow responding system. The demonstrated crystallizer design is a basis for extending the enhanced process control opportunity to other applications.

@en

A hollow fiber membrane module was assessed for its potential in assisting crystallization processes. The membrane module was characterized in the sweeping gas membrane distillation configuration considering various solution and sweeping gas flow rates, temperatures and solution concentrations. A model, coupling mass and heat transfer, was developed to predict the membrane flux. The effect of the process conditions on the membrane flux was experimentally determined and the results were used to validate the model. Feed temperature and air flow rate were found to have a significant effect on the membrane flux. Having found the optimal process conditions for membrane distillation process, batch seeded crystallization experiment were performed to confirm the potential of membrane distillation in the generation of adequate rate and level of supersaturation. Since the desired supersaturation level could be maintained in the crystallizer while seeds were growing, it is confirmed that membrane distillation can be an efficient alternative to conventional supersaturation generation processes. Finally, comparing the modelling results with experiments confirms the acceptable accuracy and predictability capability of the developed model.@en

When applied to a pure component suspension in an apolar solvent, a strong inhomogeneous electric field induces particle movement, and the particles are collected at the surface of one of the two electrodes. This new phenomenon was used to separately isolate two organic crystalline compounds, phenazine and caffeine, from their suspension in 1,4-dioxane. First, crystals of both compounds were collected at different electrodes under the influence of an electric field. Subsequent cooling crystallization enabled the immobilization and growth of the particles on the electrodes, which were separately collected after the experiment with purities greater than 91%. This method can be further developed into a technique for crystal separation and recovery in complex multicomponent suspensions of industrial processes.

@en

Salts of chiral molecules, which originally crystallize as racemic compounds, could form conglomerates. The utilization of such conglomerate salts, as intermediates for the deracemization of corresponding racemic compounds, expands the theoretical application range of Viedma Ripening by roughly 10-fold. In the present study, the use of temperature cycling on conglomerate forming salts as an alternative technique for Viedma Ripening was studied. The racemic compound Phenylalanine (Phe) was successfully deracemized via its conglomerate-forming salt with 2,5-xylenesulfonic acid (XSA) by continuous heating-cooling cycles applied to its suspension in glacial acetic acid, coupled with a solution racemization reaction. In addition, the dependence of the deracemization rate on the operational parameters was studied. Enhanced racemization reaction kinetics, either by a larger amount of free amino acid or by a higher concentration of catalyst, was shown to accelerate the deracemization process. It seems to indicate that a concentration difference between the two enantiomers, which could be diminished by a faster racemization rate, behaves as one of the major rate-limiting factors for the deracemization process. A larger mass fraction of solid dissolving and recrystallizing in the heating-cooling cycles, achieved by either a larger temperature swing or a smaller dry mass concentration, also leads to a faster deracemization. A change in cooling rate does not affect the deracemization rate significantly within the range tested, indicating a limited presence of secondary nucleation of the minor enantiomers. The results can be used as a preliminary foundation for process optimization as well as mechanisms investigation. The advantages and disadvantages of temperature cycling and Viedma Ripening, as deracemization methods in an industrial setting, are discussed.

@en

A comprehensive methodology to carry out a sequential parameter estimation approach has been developed and validated for the determination of the kinetic parameters of the crystallization of a generic organic compound. The strength of the approach lies in the thorough design of isothermal experiments which facilitate the isolation and/or decoupling of the different crystallization phenomena. This methodology has been applied for the parameter estimation of primary and secondary nucleation, growth and agglomeration kinetics. The resulting crystallization model has been able to reproduce the quantiles d₁₀, d₅₀ and d₉₀ of the volume-based particle size distribution of an independent seeded validation experiment with an error below 10 μm. The deviation in the prediction has been increased in the case of an independent unseeded experiment, although errors below the uncertainty of the measurement have been always obtained. The methodology here proposed is intended to be an efficient strategy for rapid modeling of batch crystallization processes. Â

@en