Name : Aayushi Ganeshkar

Roll No. BT16CME012

Simulated Moving bed Chromatography & its application to

chirotechnology
 REVISION
What are chiral molecules ?
In chemistry, when a molecule can have an exact mirror images of itself and be non-superimposable it
is referred to as chiral.
They share the same physical properties except their effect on the rotation of polarized light, that is,
they are optically active , and are chemically identical except when reacting with other chiral
compounds.

What are enantiomers?

Each of a pair of molecules that are mirror images of each other.

What are diastereomers?

Diastereomers are stereoisomers that are not mirror images of one another and are non-
superimposable on one another. Stereoisomers with two or more stereocenters can be diastereomers.

What is Racemic mixture ?

Racemic mixture, or racemate is one that has equal amounts of left- and right-
handed enantiomers of a chiral molecule.
 Need for Separation of Chiral molecules

 The molecules, if having a different optical activity, behave in different way, and need to be separated if want
get desired effect.

e.g. Drug Thalidomide

It is a chiral drug that was administered to pregnant women as a racemic mixture Tragically, it was realized that
only one enantiomer is beneficial; the other is believed to be responsible for major limb malformations in fetuses
and other birth defects.

 Therefore, separation of enantiomers should be done in Stage I of drug administration.

If the drug to be marketed is a pure enantiomer, this are the three main techniques for obtaining
enantiomerically pure chemicals in large quantities. The other two methods are:
(1) enantioselective chemical or enzymatic synthesis
(2) chiral resolution through derivatization and the formation of a compound that can be easily separated
(3) Chiral chromatography
But ,
(1) The development of a new method for asymmetric, enantioselective, chemical or
enzymatic synthesis is a lengthy, expensive procedure, which might result in a process
with a relatively low enantioselectivity.

(2) Chiral resolution is often labor-intensive.

Remedy :
The alternative solution to these two strategies is to use chromatographic techniques to
separate the racemic mixture obtained via non-enantioselective synthesis at all stages of
chiral drug development.This can be done using Simulated moving-bed (SMB) technology.
 Simulated Moving-Bed (SMB) Technology

 In this method, the adsorbent solid particles

flow along the column counter currently to the
fluid stream, at a velocity that renders the
propagation rates of A and B negative and
positive, respectively.
 A and B can be continuously fed in the middle
of the column.
 The concentration profiles develop, and pure
samples of A and B can be collected at the
extract and raffinate port, respectively.

But, as we have studied in IAT, all we wanted to

separate the two substances effectively is high
resolution, But graph doesn’t seem to be highly
resolved ?

Let’s understand this concept by story of The tortoise and the hare.
 The Tortoise and The hare

 This situation is similar to that of a swift hare and a sluggish turtle running a race on a straight
track. They will both reach the track, but at different times.

 If the two animals run on a moving belt, whose velocity is intermediate but opposite to theirs ,
the faster animal will fall off on the right-hand side, and the slower on the left-hand side.

 With this principle in mind, a device can be constructed that can separate cats and turtles,
which are fed continuously to the center of the moving belt, that is, a turtle-hare continuous
separator.
Working Principle

 The moving bed enables the

achievement of high purity
even if the resolution of the
two peaks is not excellent
because only the purity at the
two tails of the concentration
profiles, where the withdrawal
ports are located, is of interest.

 In principle, this moving bed

concept can be fully exploited in
the true moving- bed (TMB) unit
 Four counter current sections similar to that in the
figure are used, each playing a specific role in the
separation.

 The mixture to be separated is fed between sections 2

and 3 where A and B are separated. The more-retained
A and the less-retained B are collected in the extract
and raffinate, respectively.

 A desorbent or eluant is fed to section 1 to regenerate

the solid phase before recycling and the fluid phase is
regenerated in turn in section 4.

 However, the movement of the adsorbent particles

causes problems as a result of attrition and mixing.
Therefore, in practice this is simulated by using fixed
beds of adsorbents (chromatographic columns) and
periodically moving the inlet and outlet ports to the unit
in the same direction as the fluid flow
Advantages of SMB technology over conventional
Chromatography :

 The process is continuous, thus enabling unattended operation and stable product quality.

 The solvent requirement and the productivity per mass of the chiral stationary phase is significantly
lower

 SMB technology is significantly more robust than preparative chromatography because it requires a
smaller number of theoretical plates to achieve the same product purity.

 SMB technology can be adopted at different scales, with essentially similar equipment because only
the column size and pump performance are different.

 Although many chiral SMB separations have been developed, it is likely that some of them will never be
disclosed for proprietary reasons, so their could be many more applications than we think when it
comes to separation of chiral compounds.
Simulated Moving Bed Reactor
(SMB-R)
 A further extension of the SMB
technology involves the coupling of
the chemical reaction and the
continuous chromatographic
separation process in a single
apparatus, the SMB reactor (SMBR).
 This is a hybrid process, conceptually
similar to reactive distillation, which
is not energy-intensive and is
competitive with traditional processes
wherein the reaction and separation
are carried out in different devices.
 In reactions limited by chemical
equilibrium where more than one
product is formed, conversion can be
enhanced in this kind of hybrid
apparatus because the products are
separated as they are formed. Here,
Concluding remarks :

 Recently, SMB separation technology has been successfully

extended from hydrocarbons and sugars to fine chemicals,
particularly enantiomers.

 The driving force has been the increasing need for pure
enantiomeric products, particularly in the pharmaceutical
industry, and for the rapid and reliable development of new
chiral drugs.

 Conditions that have enabled this development to be

sustained have been the availability of an established
technology for SMB units of different scales, from grams
per day to tons per year, and stable, efficient and relatively
inexpensive chiral stationary phases.
RESEARCH
PAPERS
 Simulated moving bed chromatography is a new technology to the
pharmaceutical industry and its usefulness has yet to be established.

 To properly determine its potential, we arranged an evaluation using the

enantio-separation of a drug candidate as the example.

 A laboratory-scale system was set up and operating within two days and was
able to separate 125 g of the racemic mixture per day.

 At this throughput, each enantiomer was isolated in high purity (greater than
97,5% enantiomeric excess) and greater.
EXPERIMENTAL
SETUP
Analytical chromatography
Analysis of the product streams was carried out on a 250x4.6 mm chiralpak AD (10 ixm) column and
was eluted at 1 ml/min with 5% (v/v) 2-propanol in hexane with UV detection at 275 nm.

Choice of substrate
The separation chosen for the evaluation was that of the enantiomers of a drug candidate, which, at
that time, was proving difficult to separate by conventional crystallization methods. There was
therefore AN interest in evaluating SMB chromatography for the manufacture of the drug. The
compound chosen is a potent partial agonist at muscarinic receptors and a good analytical separation
of the enantiomers (α= 1.8) had been obtained on chiralpak AD.

Column packing
Bulk chiralpak AD (20 p~m) was packed into eight 26 mm super performance columns. Each column
contained 30 g dry mass of the stationary phase and gave a bed length of about 105 mm. For the SMB
chromatograph to work efficiently, it is important that the columns are closely matched for retention
time (<2% deviation).
Choice of mobile phase
The mobile phase used for the analytical separation on which the SMB separation was based comprised 0.1%
diethyl amine and 2% 2-propanol in hexane. To simplify product isolation, it was preferable to remove the
diethyl amine modifier and the variation of k' and c~ (determined on a 50X4.6 mm column packed with 10 Ixm
Chiralpak AD) with mobile phase composition is shown .. In consultation with Novasep, a mobile phase
containing 5% 2-propanol in hexane was selected to give a robust system.

Determination of adsorption isotherm

In order to set the SMB system parameters, it is necessary to determine the adsorption isotherms of the
components. This potentially tedious and time consuming task is greatly simplified in those cases where there
is baseline separation between the components. The software from Novasep is able to accurately model the
adsorption isotherm from the change in retention time with increased loading. One of the columns to be used
in the SMB system was set up in a conventional chromatograph (Delta-Prep 4000, Waters) and eluted with
5% (v/v) 2-propanol in hexane at 10.29 ml/min. Increasing volumes of a 100-mg/ml solution of the racemic
mixture in the mobile phase were injected and the retention times of the two peaks were measured.
OBSERVATIONS AND CONCLUSIONS
 Simulated moving bed chromatography is a new technology to the pharmaceutical industry and its
usefulness has yet to be established.

 To properly determine its potential, we arranged an evaluation using the enantio-separation of a

drug candidate as the example.

 A laboratory-scale system was set up and operating within two days and was able to separate 125
g of the racemic mixture per day.

 At this throughput, each enantiomer was isolated in high purity (greater than 97.5% enantiomeric
excess) and greater.
 Chiral considerations are now integral parts of drug research and development and of the regulatory
process. In this context, chiral chromatography has become the most important analytical tool for
determining the optical purity of organic molecules.

 Although most preparative chiral separations have been performed in the conventional batch-mode
process, interest in simulated moving-bed (SMB) chromatography is growing, because it permits large
amounts of mobile phase to be saved and productivity increased, thus reducing production costs.

 Based on the examples of three different drugs, the usefulness of this technique for the separation of
enantiomers on a preparative scale has been demonstrated. Compared to the batch elution
chromatography, a reduction of the mobile phase consumption of respectively 81% and 84% has been
achieved for the separation of the enantiomers of the anti-asthmatic agent formoterol and of the
antitussive agent guaifenesin. Furthermore, a higher throughput could be reached under SMB
conditions. The influence of feed rate and extract on the separation has also been investigated
EXPERIMENTAL
SETUP
 The feed was prepared by dissolving the racemate in the respective mobile phase. The solvents n-
heptane (99%) and ethanol were of HPLC grade.

 The chromatography was performed at 23 C using a SMB-Unit Sorbex Prep equipped with 16
columns arranged in series. Two sets of columns were used:

 (a) each column containing 5.5 g of the chiral stationary phase Chiralcel, the total volume and
amount of stationary phase was respectively 193 ml and 88 g.

 (b) each column containing 12.56 g of the chiral stationary phase Chiralcel, the total volume and
amount of stationary phase was respectively 333 ml and 201 g.

 The initial chromatographic data were determined on an analytical HPLC column filled with the
same preparative material.
In the usual batch elution chromatography the sample is injected on the top of the column and
the components separate after a certain time by moving through the column under the driving
force of the mobile phase. However, this process is not very efficient because during the
chromatography only a small part of the whole stationary phase is used for separation. A
possibility to improve the packing utilization, is given by the moving bed chromatography
principle.

According to this concept, it is not only the mobile phase which is moving but the solid-phase is
also moving. Now, for a given mobile phase rate, there is a rate of the solid-phase in the opposite
direction which should allow that the faster eluting compound (raffinate) continues to move in
the liquid direction and the slower eluting compound (extract) elutes in the opposite direction.
Under these conditions, the entire mass of the solid-phase contributing to the separation is
continuously used, thus improving considerably the productivity of the system. Obviously, this
principle is particularly suitable for a binary mixture and racemates are ideal binary mixtures.
RESULTS AND
CONCLUSIONS
On the analytical column filled with the preparative material (20 mm), as
expected the resolution and the selectivity are lower than on the column
containing the small particle size material. In order to adjust the value of
the capacity., factors to the recommended ones for performing SMB
chromatography, the mobile phase composition has been changed to
heptane–ethanol (65:35). Indeed, it is usual that an increase of the content
of the alcohol modifier causes a reduction of the capacity factors.
 INFLUENCE OF FEED RATE ON PRODUCTIVITY AND ON PURITY

In order to increase the productivity, maintaining a high purity,

the influence of the feed rate on purity has been determined,
operating at constant mobile phase rate (5.71 ml/min) and
extract rate (2.5 ml/ min).
The graph shows this influence and clearly demonstrates that
at the highest values of purity, the productivity is strongly
dependent on the desired purity.
 INFLUENCE OF EXTRACT RATE ON PURITY
We determined that the purity of both enantiomers can strongly
be influenced by variation of the extract rate . At a constant feed
rate of 0.75 ml/min and a mobile phase rate of 7.55 ml/min (cycle
time, 80 min), an increase in the extraction rate shifts the purity
of the extract to lower values whereas the purity of the raffinate
is increased.
The graph demonstrate that the productivity is strongly
dependent on the desired purity and this can be easily controlled
by finely adjusting some parameters such as the feed rate and
the extract rate.
INTRODUCTION
Simulated Moving Bed (SMB) technology is of rising interest in the
field of bio-separation. This is particularly due to its advantages
such as reduction of solvent consumption, high productivity and
final purities as well as low investment costs in comparison to
eluent chromatography.
SMB units can operate under high productivity overloaded
conditions. This leads to nonlinear competitive adsorption behavior,
which has to be accounted for when designing and optimizing new
SMB separations.
PRINCIPLE AND EXPERIMENTAL SETUP
In preparative chromatography, selectivity and efficiency no longer have the same importance
they do in analytical chromatography. A certain selectivity is required in preparative
chromatography as everywhere else in order to achieve the separation, but other parameters are
at least as important if not more so. These include the loading capacity of the stationary phase
and the maximum speed (throughput) of the process. The three main economic criteria for a large
scale separation process are:

– Productivity, i. e. the amount of product produced per unit of time and stationary phase
respectively column volume
– Eluent consumption, since this determines the cost for mobile phase in terms of preparation
and handling (tanks, water preparing systems, pumps)
– Product dilution, since this determines the cost for further product processing (e.g.,
concentration, polishing)
The classical moving bed consists of four different zones, in which different constraints
must be met:
– Zone I (between the eluent and the extract): the more firmly retained product (A,
extract) must be completely desorbed.
– Zone II (between the extract and the feed): the less firmly retained product (B,
raffinate) must be completely desorbed.
– Zone III (between the feed and the raffinate): the more firmly retained product (A,
extract) must be completely adsorbed.
– Zone IV (between the raffinate and the eluent): the less firmly retained product (B,
raffinate) must be completely adsorbed.
OPERATING CONDITIONS
The important step in designing an SMB is to find the operating conditions suited to
processing a given amount of feed per day or week or month. Since the SMB is to be used
as a preparative separation technique, which is supposed to operate under high product
overload conditions, i. e. in the non-linear part of the adsorption isotherm, the linear part of
the adsorption isotherm is normally of little importance. As mentioned previously, the
design of an SMB-separation requires the correct choice of the different flow rates of the
recycle stream, the feed stream, the eluent stream, the extract stream, the raffinate
stream and the shift period for the columns/zones, which corresponds to the simulated
“solid stream”. Other important parameters for the operating conditions are:
• The feed concentrations
• The number of columns per zone
• The column length
• The column diameter and
• The particle size.
OBSERVATIONS
In the graph below, the region of complete separation is surrounded
by three regions corresponding to three different operating regimes.
 In the region of pure raffinate, as the name indicates, the raffinate stream is
pure but the extract of pure raffinate, as the name indicates, the raffinate
stream is pure but the extract is polluted by component B.

 In the region of pure extract, the extract is pure, but the raffinate is polluted
by component A.

 In the third region (no pure fraction), both components A and B are found in
both the extract and the raffinate streams.
 The following graph illustrates the differences between operating an SMB
under linear and non-linear conditions. In particular, this figure illustrates the
effect of the overall concentration on the region of complete separation region
in the (m2, m3) plane, under conditions where all other system parameters,
such as the values of the adsorption equilibrium parameters, are kept
constant.
 Five complete separation regions plotted are shown.
 Region L corresponds to the limiting situation of a feed mixture constituted of
A and B infinitely diluted in the solvent.
 Regions 1 to 4 correspond to higher and higher feed concentrations.
CONCLUSION
Simulated Moving Bed Chromatography has been used successfully for many years on a very large
scale in the petrochemical industry. More recently, the high potential of the SMB-approach has also
been recognized by the fine chemistry and pharmaceutical industries. Applications in the
biotechnology field are increasing, where an SMB can be used putatively for many different
products and applications including, for example, proteins (enzymes), isomers, or in desalting/
polishing steps.
The SMB separates binary mixtures into the components or multi-component mixtures into two
fractions. The latter option is especially attractive if chromatographic conditions can be defined
under which the target molecule is the first or the last to elute in a multi-component mixture. Under
such circumstances the SMB can be used without technical modifications. In other situations, two
SMB systems have to be used in series.
THANK YOU