Batch preparative chromatography

There are many advantages of the SMB technology compared to batch preparative chromatography. The process is continuous and the solvent requirement is minimal compared to batch. In SMB, the whole stationary phase is used for the separation while in batch chromatography only a small part of the column is involved in the separation. This allows optimization of productivity with respect to the stationary phase. 

At the current time, there is considerable interest in the preparative applications of liquid chromatography. In order to enhance the chromatographic process, attention is now focused on the choice of the operating mode [22]. SMB offers an alternative to classical processes (batch elution chromatography) in order to minimize solvent consumption and to maximize productivity where expensive stationary phases are used. 

Buchacher et al. [43] discussed the continuous separation of protein polymers from monomers by continuous annular size exclusion chromatography. The P-CAC used for the experiments was a laboratory P-CAC type 3 as described in Table 1. The results were compared to conventional batch column chromatography in regard to resolution, recovery, fouling, and productivity. The protein used in the studies was an IgG preparation rich in aggregates. Under the conditions used, the polymers could be separated from the monomers, although no baseline separation could be achieved in either the continuous or the batch mode. The... 

In preparative chromatography, the information provided by the isotherms and by the proper model of chromatography will help the scale-up and optimization of batch and continuous purifications with minimum use of solvents and sample. Although it is rather difficult to give straightforward instructions how to optimize a preparative separation, some general guidelines are provided in this chapter. 

Preparative chromatography should be considered as enabling technology appropriate for tox/Phase 1 batches. Extensive crystallization studies, for example, to purge an impurity difficult to remove by crystallization, or to prepare an enantiomerically pure intermediate by classical resolution, may not be justified for preparing material in early drug development (Welch et ah, 2004). 

Preparative chromatography is a proven technology for the separation of specialty chemicals mainly in food and pharmaceutical industries, particularly the enantioseparation of chiral compounds on chiral stationary phases. The potential of preparative chromatographic systems were further increased by the development of continuous chromatographic processes like the simulated moving bed (SMB) process. Compared to the batch column chromatography, the SMB process offers better performance in terms of productivity and solvent consumption [2]. 

Automation allows batch chromatography to be run as a continuous process. Multiple injections using a separate pump and fraction collection provide an opportunity for continuous unattended operation. In iso-cratic separations, sample injection is often made before previously injected product elutes from the column, thus reducing cycle time and solvent consumption. Continuous and automated processes are always used with smaller columns and lower amounts of expensive enantioselective stationary phases. One of the future goals for modern PHPLC optimization would be the creation of software that would allow computer simulation modeling of nonlinear effects in preparative chromatography. 

In the last part of this book, we apply the different models discussed earlier, particularly the ideal model and the equilibrium-dispersive model, to the investigation of the properties of simulated moving bed chromatography (Chapter 17) and we discuss the optimization of the batch processes used in preparative chromatography (Chapter 18). Of central importance is the optimization of the column operating and design parameters for maximum production rate, minimum solvent use, or minimum production cost. Also critical is the comparison between the performance of the different modes of chromatography. 

Peper, S. Johannsen, M. Brunner, G. Preparative Chromatography with Supercritical Fluids - Comparison of SMB and Batch Process. Submitted for publication in J. Chromatogr. A. 2007. 

Figure 3 Fermentation modes for recombinant bacteria, yeast, and animal cells. On the left-hand side, the feed streams and harvesting streams are schematically shown for the batch, fed-batch, and continuous cultivation of microorganisms. On the right-hand side, the product concentration and the cell density are shown. For batch and fed-batch a discontinuous product concentration profile is obtained. With constitutive expression of a product, the product concentration is dependent on the cell density. Product is present in the culture supernatant during the whole production cycle and thus more susceptible to degradation. When the product formation is induction controlled, the production concentration raises sharply after addition of the inductor. The residence time of the product in the bioreactor is reduced. For continuous culture a constant product concentration profile is maintained over the entire production cycle. The residence time of the product in the bioreactor depends on the harvesting time and is shortest tor all fermentation modes when harvesting is continuously performed. Changing the mode of production can highly influence the composition of feed for preparative chromatography.

In the preparation of commercial DGEBPA, an excess of epichl orohydrin is used in order to minimize polymeriza tion of the reactants to higher molecular-weight species. Nevertheless, the typical viscous final product usually contains ca 80% by weight of the monomeric (n = 0) DGEBPA as deterrnined by gel-permeation chromatography (gpc). The manufacture of Hquid epoxy resins in a batch process has been described in some detail (9). 

HPLC separations are one of the most important fields in the preparative resolution of enantiomers. The instrumentation improvements and the increasing choice of commercially available chiral stationary phases (CSPs) are some of the main reasons for the present significance of chromatographic resolutions at large-scale by HPLC. Proof of this interest can be seen in several reviews, and many chapters have in the past few years dealt with preparative applications of HPLC in the resolution of chiral compounds [19-23]. However, liquid chromatography has the attribute of being a batch technique and therefore is not totally convenient for production-scale, where continuous techniques are preferred by far. 

Nonionic surfactants, including EO-PO block copolymers, may be readily separated from anionic surfactants by a simple batch ion exchange method [21] analytical separation of EO-PO copolymers from other nonionic surfactants is possible by thin-layer chromatography (TLC) [22,23] and paper chromatography [24], and EO-PO copolymers may themselves be separated into narrow molecular weight fractions on a preparative scale by gel permeation chromatography (GPC) [25]. 

Balannec B. and Hotier G., From batch to countercurrent chromatography , in Preparative and Production Scale Chromatography, G. Ganetsos and P.E. Barker (Editors), Marcel Dekker, New York, 1993. 

The synthesis of 6-hydroxy fluvastatin with M. rammaniana DSM 62752 gave high conversion (>95 %) in shake flask culture on 400 mL scale with 0.1 g L of fluvastatin as well as on 22 L scale in a Wave bioreactor-fed batch process at a final substrate concentration of 0.4 g L Instead of the partial purification by a second solid-phase extraction described above, 6-hydroxy fluvastatin can be obtained in high purity ( 95 %) by, for example, preparative medium-pressure liquid chromatography (MPLC) on RP18 silica gel. ... 

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