Coupling of simulated moving bed chromatography and fractional crystallisation for efficient enantioseparation

An optimised coupling of liquid chromatography and fractional crystallisation is suggested for efficient enantioseparation. As a first stage, a chromatographic separation, preferably simulated moving bed (SMB) chromatography, is applied to achieve an enantiomeric enrichment sufficient for a subsequent crystallisation. First results of the experimental and modelling work for the model system (+)-/(-)-mandelic acid in an aqueous solution are described. Chromatographic investigations involve the estimation of adsorption isotherms on a suitable chiral stationary phase and the simulation and optimisation of a corresponding SMB process. From the ternary phase diagram measured for the (+)-/(-)-enantiomer/ solvent system, the conditions required to crystallise a pure enantiomer from an asymmetric mixture can be derived. The productivity gains achievable from the combined process compared to the application of chromatography alone are discussed.     

Model-based design of a pilot-scale simulated moving bed for purification of citric acid from fermentation broth

One of the conventional processes used for the recovery of citric acid from its fermentation broth is environmentally harmful and cost intensive. In this work an innovative benign process, which comprises simulated moving bed (SMB) technology and use of a tailor-made tertiary poly(4-vinylpyridine) (PVP) resin as a stationary phase is proposed. This paper focuses on a model-based design of the operation conditions for an existing pilot-scale SMB plant. The SMB unit is modeled on the basis of experimentally determined hydrodynamics, thermodynamics and mass transfer characteristics in a single chromatographic column. Three mathematical models are applied and validated for the prediction of the experimentally attained breakthrough and elution profiles of citric acid and the main impurity component (glucose). The transport dispersive model was selected for the SMB simulation and design studies, since it gives a satisfactory prediction of the elution profiles within acceptable computational time. The equivalent true moving bed (TMB) and SMB models give a good prediction of the experimentally attained SMB separation performances, obtained with a real clarified and concentrated fermentation broth as a feed mixture. The SMB separation requirements are set to at least 99.8% citric acid purity and 90% citric acid recovery in the extract stream. The complete regeneration in sections 1 and 4 is unnecessary. Therefore the net flow rates in all four SMB sections have been considered in the unit design. The influences of the operating conditions (the flow rate in each section, switching time and unit configuration) on the SMB performances were investigated systematically. The resulting SMB design provides 99.8% citric acid purity and 97.2% citric acid recovery in the extract. In addition the citric acid concentration in the extract is a half of its concentration in the pretreated fermentation broth (feed).     

Simulated moving bed chromatography for the separation of enantiomers

Simulated moving bed (SMB) chromatography, a continuous multi-column chromatographic process, has become one of the preferred techniques for the separation of the enantiomers of a chiral compound. Several active pharmaceutical ingredients, including blockbuster drugs, are manufactured using the SMB technology. Compared to single column preparative chromatography, SMB separations achieve higher productivity and purity, while reducing the solvent consumption. The SMB technology has found applications both at small and large scales. Design methods have been developed for robust operation and scale-up, using data obtained from analytical experiments. In the last few years, rapid developments have been made in the areas of design, improved process schemes, optimization and robust control. This review addresses these developments, as well as both the fundamentals of the SMB science and technology and some practical issues concerning the operation of SMB units. Particular emphasis is placed on the consolidation of the “triangle theory”, a design tool that is used both in the academia and industry for the design of SMB processes.     

Recombinant protein purification using gradient assisted simulated moving bed hydrophobic interaction chromatography. Part II: process design and experimental validation

In the first part of this work adsorption isotherm parameters were acquired to describe the migration of recombinant streptokinase in Butyl Sepharose columns at different salt concentrations. Based on these results, a simulated moving bed (SMB) chromatographic process was designed and realised, which exploits a two-step salt gradient and allows the continuous separation of streptokinase from contaminants present in a clarified Escherichia coli cell lysate solution. This second part describes the design of the three-zone open-loop gradient SMB process applying both equilibrium theory and an equilibrium stage model and presents results of a series of experiments aiming to obtain pure streptokinase. Moreover, the potential of the SMB process and the design approach are evaluated.     

PowerFeed operation of simulated moving bed units: changing flow-rates during the switching interval

A possible way to improve the separation performance of simulated moving bed (SMB) units is to change the internal and external liquid flow-rates during the switching period. This operation mode, referred to as PowerFeed, is examined in this work through a model analysis. Similar to the Varicol process, which allows for the asynchronous movement of the ports, the PowerFeed process exhibits more degrees of freedom than the classical SMB process and therefore allows more room for optimization. Using an optimization technique based on a genetic algorithm, all three processes have been optimized for a few case studies in order to determine their relative potentials. It is found that PowerFeed and Varicol provide substantially equivalent performances, which are however significantly superior to those of the classical SMB process.     

Optimal design of batch and simulated moving bed chromatographic separation processes

Preparative chromatography, especially simulated moving bed (SMB) chromatography, is a key technology for the separation of fine chemicals on a production scale. Most of the design methods for batch and SMB processes proposed in the open literature deal with the optimisation of the operating conditions for a given chromatographic unit only. Therefore, a comparison of the process economy may lead to incorrect results. In this contribution, an effective strategy for the optimal choice of all process parameters (operation and design parameters) is proposed. The main idea of this strategy is to apply a detailed and experimentally validated process model and to reduce the number of influencing parameters by introducing and optimising dimensionless process parameters. It is shown that there is an infinite choice of design and operating parameters to achieve maximum productivity or minimum separation costs and not at the maximum pressure drop only. The detailed design of the chromatographic unit and the selection of the operating conditions can be adjusted by considering the availability of columns and packing materials. As the model system, the separation of a racemic mixture (EMD53986) on Chiralpak AD was investigated. After complete optimisation of a batch and a SMB unit, a real comparison of the process economy can be achieved. Finally, the influences of two different objective functions, productivity and specific separation cost, are analysed.     

Optimization of simulated moving bed chromatography with fractionation and feedback: Part I. Fractionation of one outlet

A novel modification of simulated moving bed (SMB) technology, referred to as fractionation and feedback SMB (FF-SMB), has been introduced recently. This concept is based on fractionating one or both outlet streams and feeding the off-spec fractions back into the unit alternatingly with the original feed mixture. In this paper, the optimization problem of FF-SMB realizing one outlet fractionation is considered. A mathematical optimization framework based on a detailed process model is presented which allows to evaluate quantitatively the potential of this operating scheme. Detailed optimization studies have been carried out for a difficult separation characterized by small selectivity and low column efficiency. The results reveal that the proposed fractionation and feedback regime can be significantly superior to the classical SMB chromatography, in terms of both feed throughput and desorbent consumption. The effect of the feeding sequence on the performance of FF-SMB is also examined.     

Optimizing control of simulated moving beds--experimental implementation

The operation of simulated moving beds (SMBs) at their optimal operating conditions is difficult and not robust. Therefore, it is common practice to operate SMB units far from their economic optimum in order to tolerate uncertainties in the system and minimize the effect of disturbances. Recently, we have proposed an on-line optimization based SMB control scheme that allows to exploit the full economic potential of SMB technology. The goal of this work is two-fold. Firstly, to experimentally evaluate and demonstrate the capability of the controller to optimize and operate the SMB units, thus delivering the products with maximum productivity and minimum solvent consumption. Secondly, to show the suitability of the controller even using a minimum of system information, thus making the detailed isotherm measurements redundant and saving time in the separation design phase. This paper reports and discusses the first experimental implementation of the control concept on a high purity separation of nucleosides (uridine, guanosine) with an eight-column four-section SMB where the species to be separated are retained on the source 30RPC stationary phase according to a linear isotherm.     

Optimization of simulated moving bed and column chromatography for a plasmid DNA purification step and for a chiral separation

This work analyzes the performance of the SMB and the column chromatography processes for two different case studies: the first stage of the plasmid DNA (pDNA) polishing, and the Tr?ger's base enantiomer separation, in which the adsorption isotherms are linear and non-linear, respectively. Simulation tools are used together with an optimization routine (Non-Sorting Genetic Algorithm (NSGA)) in order to find the optimum operating conditions leading to maximum productivity and minimum solvent consumption; the optimum solution for each of the processes is a curve on the productivity-solvent consumption plane, the so-called Pareto set. The comparison between the column and the SMB processes is based on the relative position of the two Pareto sets calculated at equal conditions and for the same final purity and recovery of the target species. The results show that SMB is superior to column chromatography in the two case studies investigated, i.e. in the case of the linear isotherm (pDNA), the productivity gain is up to a factor two for a given value of the solvent consumption. Furthermore, the flexibility of the SMB operation is larger, since the Pareto sets are flatter and they prolong into regions of the productivity-solvent consumption plane that are not accessible with the column chromatography process.     

Optimization of startup and shutdown operation of simulated moving bed chromatographic processes

This paper presents new multistage optimal startup and shutdown strategies for simulated moving bed (SMB) chromatographic processes. The proposed concept allows to adjust transient operating conditions stage-wise, and provides capability to improve transient performance and to fulfill product quality specifications simultaneously. A specially tailored decomposition algorithm is developed to ensure computational tractability of the resulting dynamic optimization problems. By examining the transient operation of a literature separation example characterized by nonlinear competitive isotherm, the feasibility of the solution approach is demonstrated, and the performance of the conventional and multistage optimal transient regimes is evaluated systematically. The quantitative results clearly show that the optimal operating policies not only allow to significantly reduce both duration of the transient phase and desorbent consumption, but also enable on-spec production even during startup and shutdown periods. With the aid of the developed transient procedures, short-term separation campaigns with small batch sizes can be performed more flexibly and efficiently by SMB chromatography.     

Pulsed flow modulation two-dimensional comprehensive gas chromatography-tandem mass spectrometry with supersonic molecular beams

Pulsed flow modulation (PFM) two-dimensional comprehensive gas chromatography (GC x GC) was combined with quadrupole-based mass spectrometry (MS) via a supersonic molecular beam (SMB) interface using a triple-quadrupole system as the base platform, which enabled tandem mass spectrometry (MS-MS). PFM is a simple GC x GC modulator that does not consume cryogenic gases while providing tunable second GC x GC column injection time for enabling the use of quadrupole-based mass spectrometry regardless its limited scanning speed. The 20-ml/min second column flow rate involved with PFM is handled, splitless, by the SMB interface without affecting the sensitivity. The combinations of PFM GC x GC-MS with SMB and PFM GC x GC-MS-MS with SMB were explored with the analysis of diazinon and permethrin in coriander. PFM GC x GC-MS with SMB is characterized by enhanced molecular ion and tailing-free fast ion source response time. It enables universal pesticide analysis with full scan and data analysis with reconstructed single ion monitoring on the enhanced molecular ion and another prominent high mass fragment ion. The elimination of the third fragment ion used in standard three ions method results in significantly reduced matrix interference. GC x GC-MS with SMB improves the GC separation, and thereby our ability for sample identification using libraries. GC-MS-MS with SMB provides better reduction (elimination) of matrix interference than GC x GC-MS. However, it is a target method, which is not always applicable. GC x GC-MS-MS does not seem to further reduce matrix interferences over GC-MS-MS and unlike GC x GC-MS, it is incompatible with library identification, but it is beneficial to have both GC x GC and MS-MS capabilities in the same system.     

Optimal operation of simulated moving bed chromatographic processes by means of simple feedback control

In this contribution, simple methods are presented for controlling a simulated moving bed (SMB) chromatographic process with standard PI (proportional integral) controllers. The first method represents a simple and model-free inferential control scheme which was motivated from common distillation column control. The SMB unit is equipped with UV detectors. The UV signals in the four separation zones of the unit are fixed by four corresponding PI controllers calculating the ratio of liquid and solid flow in the respective separation zone. In order to be able to adjust the product purity a second, model-based control scheme is proposed. It makes use of the nonlinear wave propagation phenomena in the apparatus. The controlled chromatographic unit is automatically working with minimum solvent consumption and maximum feed throughput--without any numerical optimization calculations. This control algorithm can therefore also be applied for fast optimization of SMB processes.     

Liquid chromatography mass spectrometry with supersonic molecular beams

A new approach for liquid chromatography mass spectrometry (LC-MS) is described, based on achieving soft thermal vaporization followed by supersonic expansion and direct sample compound ionization, while in a supersonic molecular beam (SMB). The soft molecular vaporization step utilizes spray formation that is continued by fast thermal vaporization inside a channel supersonic nozzle, followed by ultrafast supercooling in a supersonic expansion. The short time (several microseconds) spent by the vaporized compound in the heated nozzle prior to its expansion cooling may result in incomplete vibrational equilibrium and thus reduced degree of dissociation. In addition, even if vibrational equilibrium at the nozzle temperature is obtained, the sample compounds have significantly reduced time for their dissociation, which is thus further minimized (kinetic consideration). As soon as the molecules expand and form a SMB, they are supercooled and any further dissociation is avoided. While in the SMB, the sample molecules can be ionized either by electron ionization as described in this paper or by hyperthermal surface ionization. The major goal of this method is to obtain high quality library searchable electron ionization mass spectra, for a broad range of thermally labile compounds, with higher sensitivity than that achievable by particle beam LC-MS. The soft thermal vaporization nozzle is described and mass spectral results with corticosterone are demonstrated. The potential advantageous features of this new method are discussed.     

Parametric uncertainties and influence of the dead volume representation in modelling simulated moving bed separation processes

In this study, a systematic numerical procedure for identifying the model parameters of simulated moving bed (SMB) separation processes is developed. The parameters are first estimated by minimizing a weighted least-squares criterion using experimental data from batch experiments, e.g. the time evolution of the concentration of elution peaks. Then, a cross-validation is achieved using data from experiments in SMB operation. At this stage, the importance of a careful modelling of the dead volumes within the SMB process is highlighted. In addition, confidence intervals on the estimated parameters and on the predicted concentration profiles are evaluated.     

Enantioseparation of a chiral epoxide by simulated moving bed chromatography using Chiralcel-OD

Summary The feasibility of using simulated moving bed (SMB) chromatography for the chiral separation of a racemic epoxide with Chiralcel-OD as the stationary phase is demonstrated on a semi-preparative scale. Operating conditions for the separation are chosen with the help of a simple chart that depicts visually the interrelationships between the system flow rates and the SMB design criteria. The 12 column (each 100 mm×16 mm ID) SMB system continuously resolved the racemic mixture at a rate of 11.5 g/24 hr into streams with 95% and 94.4% e.e. (enantiomeric excess). A comparison of the SMB process with an optimized multiple-injection conventional chromatographic separation showed similar specific production rates for both methods, but a seven-fold lower solvent consumption for the SMB.     

Intermittent simulated moving bed chromatography: 3. Separation of Tröger's base enantiomers under nonlinear conditions

One of the modified simulated moving bed (SMB) processes, the intermittent SMB (I-SMB) process, has been recently analyzed theoretically [1] and its superior performance compared to the conventional SMB process has been demonstrated at a rather low total feed concentration through experiments and simulations [2]. This work shows that the I-SMB process outperforms the conventional SMB process also at high feed concentration where the species are clearly subject to a nonlinear adsorption isotherm. In the case of the separation of the Tröger's base's enantiomers in ethanol on ChiralPak AD, the two processes operated in a six-column 1-2-2-1 configuration (one column in sections 1 and 4 and two columns in sections 2 and 3) and in a four-column 1-1-1-1 configuration (one column in each section) are compared at high feed concentration through both experiments and simulations. Even under nonlinear conditions the four column I-SMB process can successfully separate the two enantiomers achieving purity levels as high as the two six column processes and exhibiting better productivity.     

Recombinant protein purification using gradient-assisted simulated moving bed hydrophobic interaction chromatography. Part I: selection of chromatographic system and estimation of adsorption isotherms

The design of gradient simulated moving bed (SMB) chromatographic processes requires an appropriate selection of the chromatographic system followed by the determination of adsorption isotherm parameters in the relevant range of mobile phase conditions. The determination of these parameters can be quite difficult for recombinant target proteins present in complex protein mixtures. The first part of this work includes the estimation of adsorption isotherm parameters for streptokinase and a lumped impurity fraction present in an Escherichia coli cell lysate for a hydrophobic interaction chromatography (HIC) matrix. Perturbation experiments were carried out using a Butyl Sepharose matrix with purified recombinant protein on buffer equilibrated columns as well as with crude cell lysate saturated columns. The Henry constants estimated for streptokinase were found to exhibit in a wide range a linear dependence on the salt concentration in the mobile phase. These parameters were applied in subsequent investigations to design a simulated moving bed (SMB) process capable to purify in a continuous manner recombinant streptokinase from the E. coli cell lysate.     

Simulated moving bed technology: old and new

The Simulated Moving Bed (SMB) concept has been applied to the separation of different mixtures as a continuous counter current separation process, avoiding several problems related with solid motion. The aim of this work is to present some relevant examples of SMB separations corresponding to the two major ages in the use of the SMB concept, here named “old” and “new” applications. The “old” applications of SMB technology in the petrochemical industry are still important, with large and highly productive units; and the “new” applications of the second “age” of SMB concept are from the fine chemical, pharmaceutical and biochemistry areas, associated with the demand of high purity products during the last 10 years. Different examples are presented for different ages: a UOP Parex ^® process for the “old”, modelled with the equivalent True Moving Bed (TMB) approach; and a chiral resolution for the “new”, modelled by the real SMB model. Some of the latest developments are also mentioned: the non conventional techniques as the Varicol ^® process, PowerFeed, Modicon, M3C or Enriched Extract-SMB (EE-SMB), MultiFeed (MF), Outlet Streams Swing (OSS) or Pseudo-SMB, involving considerable changes in the SMB concept itself. The use of the last optimization/modelling packages for the development of design techniques, either at the conception stage as well as for performance improvements of existing units is emphasized.     

Solution crystallisation via a submerged liquid-liquid phase boundary: oiling out

In many situations the process of crystallisation from solution is known to occur via metastable crystalline states (polymorphs or solvates). Here we present what we believe to be a novel example of small molecule crystallisation in which the initial separation of a solute rich liquid phase precedes the crystallisation event. We believe this occurs because a submerged liquid-liquid phase boundary is accessible within the metastable zone of the crystal nucleation process.     

Intermittent simulated moving bed chromatography: 2. Separation of Tröger’s base enantiomers

The intermittent simulated moving bed (I-SMB) process is a modification of the conventional SMB process that has been recently analyzed theoretically [1]. Here, we present a comparative analysis of the two processes, each operated in a six column 1-2-2-1 configuration (one column in sections 1 and 4 and two columns in sections 2 and 3) and in a four-column 1-1-1-1 configuration. Experiments are carried out on a properly modified laboratory unit to separate racemic mixtures of the enantiomers of Tröger’s base in ethanol on ChiralPak AD at a total feed concentration of 1 g/L. Simulations are carried out for the same system using the equilibrium dispersive model and a bi-Langmuir isotherm, whose parameters have been preliminarily estimated from pulse and breakthrough experiments. Experiments and simulations are fully consistent and demonstrate that the four-column I-SMB process (but not the four-column SMB process) can separate the two enantiomers at very high purity and achieve a productivity twice as large as that of the six-column I-SMB and conventional SMB processes with the same solvent consumption.