Radleys Carousel Work-Up Station User manual

Instructions
Carousel
Work-Up Station™
Cat No. RR99830
Your Local Distributor
February 2023 | Issue 5
Radleys Shire Hill, Saffron Walden, Essex, CB11 3AZ. United Kingdom.
Tel: [phone]
Email: [email protected]m
Web: www.radleys.com

Page 1
Index
Page 1 Index
Page 2 Introduction
Warranty
Page 3 Safety Information
Important WARNINGS
Page 4 Products & Accessories
Page 5 Carousel Work-Up Set-Up & Operation
Page 6 Using the SpeediFlow Booster
Page 7 Using the SpeediFlow Booster - continued
Page 8 Dismantling the SpeediFlow Booster
Page 9 Choice of Columns
Page 10 Sample Preparation Prior to SPE Purification
Using 20µm PE Filtration Columns
Page 11 Using Phase Separation Columns
Page 12 Using Liquid/liquid Extraction Columns
Page 13 Selecting the Correct SPE Column
Page 14 Guidelines for Using SPE Columns
Page 15 Guidelines for Using SPE Columns - continued
Page 16 WARRANTY - EMAIL BAC
RR99830 - Carousel Work-Up Station

RR99830 - Carousel Work-Up Station
Thank you for purchasing your Carousel Work-Up Station
Please read this Instruction Manual thoroughly before using this apparatus.
Introduction
The Carousel Work-Up Station is designed to make your post synthesis work-up quick and easy,
avoiding downstream bottlenecks...
The Carousel Work-Up Station accepts up to 12 x 70ml columns loaded into one of two identical stackable racks.
The lower rack supporting 12 corresponding Carousel Reaction Tubes or standard 1 inch boiling tubes
for subsequent sample collection.
The Carousel Work-Up Station allows up to two levels of 70ml columns stacked one above the other. This enables parallel
and/or sequential work-up in 12 samples, using filtration, phase separation or solid phase extraction techniques.
•Designed for use with the 12 position Carousel Reaction Station or similar parallel synthesisers.
•Accepts 12 x Carousel Reaction Tubes or standard 1 inch boiling tubes
(maximum tube dimensions 29mm OD x 150mm long)
•Accepts 70ml columns; including 20µm Filtration, Phase Separation, Liquid/Liquid Extraction, Silica, SCX, Aminopropyl
and SAX Acetate (maximum column dimensions 29mm OD x 155mm long to tip)
•Unique design uses no taps, valves or drip needles, making assembly and operation fast and simple,
minimising cleaning and consumable costs.
•Vacuum free operation also makes removal of collections tubes easy, and prevents drying of columns.
•Use under gravity or with innovative SpeediFlow flow booster, allowing precise individual control of solvent
flow within each column.
•The unique SpeediFlow is a hand held pressuriser for easy insertion into columns. It features a mechanically expanded
Viton O-ring, operated by a trigger with integral locking device, for unsupported gas tight operation.
•SpeediFlow features a safety relief valve that vents at 6 to 8psi, preventing accidental over-pressure.
•Ergonomically designed, lightweight aluminium racks are easy to load with collection tubes or work-up columns.
They are exceptionally stable, with good access and visibility of all columns and tubes.
•Identical racks lock together for ease of operation or transportation from location to location.
•An optional 3rd rack can be stacked above the 2nd rack for sequential work-up.
Warranty
The Carousel Work-Up includes one-year full parts and labour warranty from date of original purchase.
Warranty will only be valid if a completed Warranty email back is returned within 1 month
of date of purchase (see last page).
In the event of product failure please contact your local distributor.
Please do not return any goods without prior agreement.
Page 2

Safety Information
The following symbols are intended to assist the user in the safe and efficient
operation of the Carousel Work-Up Station.
Important WARNINGS
Please read these instructions completely before using your Carousel Work-Up Station
Only operate in a fume cupboard with protective safety sash.
When using chemicals and reagents always follow appropriate
safety instructions and chemical handling procedures.
If used in conjunction with the SpeediFlow please ensure that it is connected to an air
or gas line with an INLET PRESSURE THAT DOES NOT EXCEED 6 psi (approx. 0.4 bar)
and that the tubing is correctly secured so it cannot blow off under pressure.
RR99830 - Carousel Work-Up Station
Page 3
Warning
Applies when there is a possibility
of personal injury.
Important Note
Alerts the user to important facts.
!
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Important, Important,
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Important Note
The Carousel Work-Up Station should only be operated by
trained and competent personnel. As with all procedures, care
should be taken to monitor your work-up at all stages.
Important, Important,
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Page 4
RR99830 - Carousel Work-Up Station
Products & Accessories
Cat No Description Pk Qty
Carousel Work-Up Station
RR99830 Carousel Work-Up Station (Two racks and SpeediFlow) 1
Carousel Work-Up Station Components & Accessories
RR99831
RR99850
1
RR99852
1
1
RR99856 1
RR99858 1
RR99860 5
RR99861 2
RR99862 5
RR99864 5
RR99866 5
RR99868 5
RR99870 16
RR99871 16
RR99872 16
RR99873 16
RR99874 50
RR99875 50
RR99877 50
RR99878 50
RR99840
Carousel Work-Up Rack
SpeediFlow (includes Adapter Head for 70ml Columns
SpeediFlow Adapter Head for 15ml Columns
SpeediFlow Adapter Head for 150ml Columns
SpeediFlow Adapter Head for AutoCup
Replacement Viton O-ring - for 70ml Columns
Replacement Perlast O-ring - for 70ml Columns
Replacement Viton O-ring for 15ml Columns
Replacement Viton O-ring for 25ml Columns
Replacement Viton O-ring for 150ml Columns
Replacement Viton O-ring for AutoCup
Silica 10g/70ml
Aminopropyl 10g/70ml
SCX 10g/70ml
SAX Acetate 10g/70ml
20um PE Filtration 70ml
Phase Separation 70ml
Liquid/Liquid Extraction, 10ml Sample, 70ml
Liquid/Liquid Extraction, 20ml Sample, 70ml
SpeediFlow as Connection Tubing - 5m length 1
Carousel Work-Up Station Systems:- include all the important components and accessories
required for efficient operation
Cat No Description Pk Qty
RR99876 - Carousel Work-Up Column Pack
Comprising of the following:
RR99870 Silica 10g/70ml 16
RR99871 Aminopropyl 10g/70ml 16
RR99872 SCX 10g/70ml 16
RR99873 SAX Acetate 10g/70ml 16
RR99874 20um PE Filtration 70ml 50
RR99875 Phase Separation 70ml 50
RR99880 - System 25 - Carousel Work Up Station
Comprising of the following:
RR99830 Carousel Work-Up Station (Two racks and SpeediFlow) 1
RR99874 20um PE Filtration 70ml 50
RR99875 Phase Separation 70ml 50
RR98062 Threaded lass Reaction Tube 24mm x 150mm 12
RR99840 SpeediFlow as Connection Tubing - 5m length 1
RR99881 - System 26 - Carousel Work Up Station
Comprising of the following:
RR99830 Carousel Work-Up Station (Two racks and SpeediFlow) 1
RR99831 Carousel Work-Up Rack 1
RR99870 Silica 10g/70ml 16
RR99874 20um PE Filtration 70ml 50
RR99875 Phase Separation 70ml 50
RR98062 Threaded lass Reaction Tube 24mm x 150mm 12
RR99840 SpeediFlow as Connection Tubing - 5m length 1

The Carousel Work-Up Station consists of two identical racks that stack on top of
each other. The racks lock together with a pin so that the stack of two racks can
be easily carried or moved safely and securely. In normal use the upper rack will
contain filtration, phase separation, liquid/liquid extraction or solid phase
extraction (SPE) columns, and the lower rack will hold Carousel Reaction Tubes
or standard 1 inch boiling tubes (maximum tube dimensions 29mm OD x 150mm
long) for fraction collection.
The SpeediFlow Booster is used to apply pressure to the columns to increase
flow rates. Pressure is not usually required for phase separation or liquid/liquid
extraction.
For sequential operations e.g. filtration or phase separation followed by SPE it
is possible to stack an additional rack above the other two. The crude solution is
then transferred directly from either the filtration, phase separation or liquid/liquid
extraction columns to the SPE columns in the middle level. After removal of the
upper level the SPE columns are eluted and individual fractions collected in the
lower level.
Where it is desired to collect multiple fractions, processing a single SPE
column and moving it from one position to the next and collecting sequential
fractions can achieve this most easily. In this way up to 12 fractions can be
collected in a single rack.
RR99830 - Carousel Work-Up Station
Carousel Work Up Set-up & Operation
Page 5
The Carousel Work-Up Station may be used in several ways. The initial set up
procedure is always the same.
Initial Setting Up
1. Place the appropriate number of collection tubes into a rack. Collection tubes
may be Carousel Reaction Tubes or 1 inch boiling tubes.
2. To locate a second rack on top of the first, gently pull out the the spring loaded
knob and rotate it by 90˚. Then place it it onto the lower rack so that the
handle on the lower rack fits into the the slot in the upper rack. The racks will
only fit in one orientation, with the knobs facing forwards. Then gently rotate
the knob until it clicks into place. The two racks are now securely locked
together.
3. Select the appropriate tubes or columns (filtration, phase separation,
liquid/liquid extraction or SPE) corresponding to the number and position
of the collection tubes and then drop them into the correct positions.

The SpeediFlow is used to apply top (positive) pressure to Filtration or SPE
Columns. Flow rates through these columns (particularly SPE) can be slow with-
out additional pressure being applied either via vacuum or top pressure. Pressure
is not normally required for phase separation or liquid/liquid extraction and actual-
ly may cause the aqueous phase to be forced through the hydrophobic frit or the
bed of diatomaceous earth.
RR99830 - Carousel Work-Up Station
Using the SpeediFlowBooster
Important Note
Applying excessive top pressure to a phase separation column or
liquid/liquid extraction column may force the aqueous phase
through.
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The SpeediFlow is designed to be hand held for easy insertion into columns, but
has a locking device to enable unsupported operation. The SpeediFlow fits easily
into the top of a standard 70ml plastic column.
1. Connect appropriate tubing from air or nitrogen line to connector on the top
of the SpeediFlow, ensuring the tubing is secure
2. Insert SpeediFlow nozzle into the plastic column.
3. Squeeze trigger lever - initially the O-ring will expand and form a gas tight seal
within the column. As the O-ring seals on the column inner a ‘witness line’ will
appear indicating a good all round seal.
4. Squeezing of the trigger lever simultaneously opens a valve within the handle
that allows gas to flow into the column, creating a postive pressure to increase
the flow rate.
Warning
The SpeediFlow should be connected to an air or nitrogen line with
an inlet pressure not exceeding 6 psi (approx. 0.4 bar) and the
tubing secured so that it will not blow off under pressure.
!
Warning
If the safety relief valve starts to vent - then reduce incoming gas
pressure to below 6 psi or until it ceases.
REMOVALORTAMPERINGWITHTHESAFETYRELIEFVALVEMAYLEADTOPERSONALINJURY
AND WILLINVALIDATEYOURWARRANTY.
!
Page 6
Important Note
The SpeediFlow should only be operated by trained and competent
personnel. As with all procedures, care should be taken to monitor
your work-up at all stages.
Important, Important,
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Important Note
The SpeediFlow features a safety relief valve which vents at 6 to
8psi, preventing accidental over-pressure.
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Using the SpeediFlow Booster - continued
RR99830 - Carousel Work-Up Station
Page 7
Insert SpeediFlow
nozzle into the
column.
Squeeze trigger
and press the thumb
button until it locks.
Release trigger.
O-ring now forms
a gas tight seal
within the column.
In this position,
a constant pressure
is maintained in the
column without
holding the trigger.
as will now flow
into the column
speeding up the flow
of your sample.
To release, squeeze
trigger. The thumb
button will release
and the SpeediFlow
may be withdrawn.
5. If you want to leave the SpeediFlow unsupported then press the thumb
button when the trigger lever is fully compressed. This will lock the trigger into
position and and allow a constant pressure to be maintained in the column
without holding the trigger.
6. To release the SpeediFlow simply squeeze the trigger lever - the thumb button
will release and the SpeediFlow may be withdrawn.
Warning
The SpeediFlow is a pressurised device and therefore is not
designed for unattended operation.
!
SpeediFlow Adapter Heads
The SpeediFlow™now includes an expanded range of optional adapters to fit
a wide range of standard column sizes...
For use with 15ml, 25ml, 70ml and 150ml columns plus the Whatman AutoCup®.
Thus allowing the SpeediFlow to be used in other SPE
or filtration applications.
Brings added versatility to the SpeediFlow.
150ml Whatman
AutoCup®
70ml 15ml
150ml
Whatman
AutoCup®
70ml
15ml

RR99830 - Carousel Work-Up Station
Page 8
The SpeediFlow is designed so that the nozzle can be removed for cleaning
and to allow easy replacement of the O-ring. This can be carried out following the
procedure below:
1. Pull out the thumb button. The trigger lever will now fall away from the handle.
2. The nozzle and O-ring can now be pulled out for cleaning or replacement.
3. To re-insert the nozzle ensure that the trigger lever is fully open. Place O-ring
over nozzle and then push the nozzle and O-ring assembly into
the SpeediFlow.
4. The trigger lever may then be squeezed until it clicks into place past
the thumb button.
Warning
REMOVALORTAMPERINGWITHTHESAFETYRELIEFVALVE
MAYLEADTOPERSONALINJURYAND WILLINVALIDATE
YOURWARRANTY.
!
Important Note
Always check the O-ring before use. If O-ring is cracked or showing
evidence of wear or chemical attack it should be replaced.
Important, Important,
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Avoid continuous immersion of the O-ring in solvents as this may
cause swelling and affect its performance and effectiveness
of the seal.
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Warning
Only operate SpeediFlow in a fume cupboard with protective safety
sash. When using chemicals and reagents always follow
appropriate safety instructions and chemical handling procedures.
!
Warning
The SpeediFlow should not be used with glass columns or vessels.
!
The SpeediFlow is supplied with a Viton®O-ring as standard.
An optional Perlast®O-ring is available for use with more aggressive solvents.
Perlast is a perfluoroelastomer (similar to PTFE) that offers excellent resistance
to a broad range of commonly used chemicals and exhibits minimal swelling even
with prolonged exposure to vapours.
Dismantling the SpeediFlow Booster

The Carousel Work-Up Station is designed to accept 70ml tubes and
columns which is the most appropriate size for most work-ups on the
Carousel Reaction Station scale.
The choice of column depends on many factors including:
• What is it you wish to achieve (filtration, purification etc)?
• The properties of the desired compound
• The nature of the sample
• The nature of the impurities
• The degree of purity required
Four types of purification can be performed...
1. Filtration of a resin or other solids use a 20µm PE Filtration Column. Using
the pressuriser to apply pressure will greatly increase the filtration rate.
2. Phase Separation columns have a 5µm PTFE hydrophobic frit and are
suitable when the organic phase is both miscible and denser than water.
3. Liquid/Liquid Extraction columns use a modified form of diatomaceous earth
to adsorb and retain the aqueous phase and are suitable when the organic
phase is either less dense or more dense than water.
4. Solid Phase Extraction columns depends on the properties of the compound
you are trying to purify and the nature of the impurities.
For sequential operations e.g. filtration or phase separation followed by SPE it
is possible to stack an additional rack above the other two. The crude solution is
then transferred directly from either the filtration, phase separation or liquid/liquid
extraction columns to the SPE columns in the ‘middle rack’. After removal of the
‘upper rack’ the SPE columns are eluted and individual fractions collected in the
‘lower rack’.
Where it is desired to collect multiple fractions, processing a single SPE
column and moving it from one position to the next and collecting sequential
fractions can achieve this most easily. In this way up to 12 fractions can be
collected in a single rack.
Choice of Columns
RR99830 - Carousel Work-Up Station
Page 9

Sample Preparation Prior to SPE Purification
Prior to purification by SPE (Solid Phase Extraction) the crude reaction mixtures,
from for example a parallel set of reactions in the Carousel Reaction Station, may
need to be worked up.
Initial work-up may involve a filtration, a phase separation and/or a liquid/liquid
extraction. For these sequential operations it is possible to stack an additional
rack above the other two. The crude solution is then transferred directly from
either the filtration, phase separation or liquid/liquid extraction columns to the SPE
columns in the ‘middle rack’. After removal of the ‘upper rack’ the SPE columns
are eluted and individual fractions collected in the ‘lower rack’.
RR99830 - Carousel Work-Up Station
Using 20µm PE Filtration Columns
Typical applications include:
• Isolation of precipitates or crystallites from the reaction mixture
• Filtration of resins prior to cleavage
• Filtration of solid supported reagents post synthesis
• Removal of solid impurities prior to SPE
Pressure can be applied to an individual column using the SpeediFlow to increase
the flow rate, particularly if the column is slow or becomes blocked. Solid
impurities, e.g. salts may be removed by using filtration columns prior to SPE.
Using Filtration Columns
1. Decant material to be filtered into the filtration column and rinse in residues.
2. Allow gravity to let material flow through or use SpeediFlow by sliding it into
top of column and applying pressure until frit is dry. Further solvent may be
added as necessary to wash any solid.
3. If the desired material is in the solution this may then be evaporated in e.g.
a enevac vacuum centrifuge of further purified via SPE for example.
Cat No Description Pk Qty
Filtration Columns
RR99874 20µm PE Filtration 50
Phase Separation Columns
RR99875 Phase Separation 50
Liquid/liquid Extraction Columns
RR99877 Liquid/liquid Extraction, 10ml Sample, 70ml 50
RR99878 Liquid/Liquid Extraction, 20ml Sample, 70ml 50
Page 10

RR99830 - Carousel Work-Up Station
Page 11
Using Phase Separation Columns
The Phase Separation Columns contain a hydrophobic frit and are designed
to separate organic phases that are denser than water (e.g. dichloromethane).
They can also be used for sequential purification, e.g. phase separation
followed by SPE.
Following partition of a crude reaction mixture between, for example dichloro-
methane and water, the mixture is transferred to the phase separation column
where the lower organic layer will run through the hydrophobic frit leaving the
upper aqueous layer in the column for disposal.
It should not be necessary to apply pressure, indeed doing so can allow the
aqueous layer to break through the hydrophobic frit.
Important Note
Phase Separation Columns must only be used with non-water
miscible solvents that are denser then water e.g. dichloromethane.
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The two phase solution should be well stirred or shaken in, for
example a Carousel Reaction Tube before transferring it to the
phase separation column.
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Important Note
The presence of even small quantities of some water miscible
solvents (e.g. ethanol, DMF) can cause the phase separation
columns to fail. In this case ensure that the water miscible solvent
is completely evaporated before partitioning it between the organic
and aqueous phases.
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RR99830 - Carousel Work-Up Station
Using Liquid/liquid Extraction Columns
The Liquid/liquid Extraction Columns contain a modified diatomaceous earth
which efficiently absorbs water from aqueous samples. The diatomaceous earth is
chemically inert and stable over the ph range 1-13.
Unlike the Phase Separation Columns the Liquid/liquid Extraction Columns can
be used with non-water miscible solvents that are either more or less dense than
water. When a two phase mixture is applied to the column the aqueous phase is
adsorbed onto the diatomaceous earth and the organic phase runs through the
column. The high surface area at the interface between the organic and aqueous
phases increases efficiency and eliminates the possibility of emulsion formation.
Further elution with more solvent elutes the organic compounds and the aqueous
phase is retained on the column for disposal.
Column Selection
Selection of the correct column is based on the volume of aqueous present in the
two phase sample. Always use a column of equal or greater capacity than the
volume of the aqueous phase. Aqueous volumes and suggested elution volumes
are given below:
Maximum Aqueous Suggested Column
Cat No Sample Volume Elution Volume Volume
RR99877 10ml 24ml 70ml
RR99878 20ml 40ml 70ml
Using Liquid/liquid Extraction Columns
1. Ensure that the two phase mixture has been thoroughly mixed by shaking
or stirring before applying to the Liquid/liquid Extraction Column.
2. Apply the two phase sample to the Liquid/liquid Extraction Column. No more
than the maximum aqueous sample volume for each column configuration
should be applied to the column - see guidelines above.
3. Wait for 3 - 5 minutes.
4. Apply a suitable water immiscible organic solvent (see table above for
appropriate elution volumes) and collect the eluant.
Page 12
Warning
Do not overload the column. This could lead to breakthrough of the
aqueous sample and contamination of the extract. It is not normally
necessary to apply pressure which may also cause breakthrough
of the aqueous phase.
!
Important Note
Do not use a solvent for elution that contains more than 10% water
miscible component as this could lead to column overload and
extract contamination.
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RR99830 - Carousel Work-Up Station
Page 13
Selecting the Correct SPEColumn
RDT offer four types of SPE column that will cover the vast majority of
purifications for the Carousel Work-Up Station. These are:
The following guidelines are intended to help in the initial selection of a column
according to the main interactions of the solid phase, polar, non-polar or
anion/cation exchange.
The characteristics of the various sorbents are described below.
Silica
The main interaction is polar via the silanol functionality. Most useful for purifying
moderately polar compounds where the impurities are either or both very non-
polar (eluted first) or very polar (retained on column).
Aminopropyl
Normally used as a polar sorbent. The amino group can hydrogen bond with a
variety of polar groups, e.g. hydroxyl, amino etc and will have polar interactions
with polar groups. It will also function as a weak anion exchanger when
protonated.
SCX
A strong cation exchanger used to extract basic compounds capable of holding a
positive charge. Can be used for extracting basic impurities from non-basic
products (which pass straight through), or for extracting basic products from non-
basic impurities (which pass straight through). The basic products can then be
eluted using a medium with a higher pH.
SAX Acetate
A strong anion exchanger used to extract compounds capable of carrying a
negative charge (e.g. weak acids). Can be used for extracting acidic impurities
from non-acidic products (which pass straight through), or for extracting acidic
products from non-acidic impurities (which pass straight through). The acidic
products can then be eluted using a medium with a lower pH.
Cat No Description Volume Pk Qty
RR99870 Silica 10g/70ml 16
RR99871 Aminopropyl 10g/70ml 16
RR99872 SCX 10g/70ml 16
RR99873 SAX Acetate 10g/70ml 16

RR99830 - Carousel Work-Up Station
Guidelines for Using SPE Columns
The overall purification process normally consists of 5 steps:
1. Sample Preparation – initial work-up
See above for guidelines on the use of Filtration, Phase Separation and
Liquid/liquid Extraction Columns.
2. SPE Column Solvation/Equilibration
SPE columns must be solvated properly before the sample is loaded.
Polar sorbents (Silica and aminopropyl (see Note)) are usually
solvated/equilibrated using the first elution solvent e.g. hexane (at least 1 column
volume (CV)). Use e.g. a multi-channel pipettor to apply the solvent to the
columns and do not allow the columns to dry out.
Ion exchange sorbents (SCX, SAX acetate and aminopropyl (see Note)) are
solvated and equilibrated with at least 1 CV of a polar solvent e.g. methanol,
ethanol or acetonitrile that is also used as the first eluting solvent.
Note: Aminopropyl may be used as a polar sorbent or weak anion exchanger. To
use as the former it should be solvated with a non-polar solvent (e.g. hexane) and
eluted with a more polar solvent (e.g. hexane/ethyl acetate). In a polar
solvent e.g. methanol or aqueous methanol it will act as a weak anion exchanger
when amino groups are protonated and the column will retain molecules that
carry a negative charge (e.g. strong acids).
3. Sample Loading
Samples can be loaded onto the columns a pipettor or by pouring directly from
the reaction tube.
For polar sorbents (Silica, Aminopropyl) load the sample in a non-polar solvent.
If the crude reaction mixture is not soluble in this add sufficient of a more polar
solvent (e.g. ethyl acetate) to achieve solution. The ratio of crude sample
adsorbed to sorbent should not exceed 1:20 for good separation.
SCX has a maximum loading of 0.3mmol/g. For a singly charged molecule
of MW 300 this is equal to 45mg on the 500mg columns, 90mg on the 1.0g
columns or 900mg on the 10.0g columns.
SAX Acetate has a loading of 0.6mmol/g. For a singly charged molecule of MW
300 this is equal to 90mg on the 500mg columns, 180mg on the 1.0g columns or
1.8g on the 10.0g columns.
Aminopropyl is useful for the extraction of e.g. strong acids which bear a
permanent negative charge. Aminopropyl has a loading of 0.6mmol/g. For a singly
charged molecule of MW 300 this is equal to 90mg on the 500mg columns,
180mg on the 1.0g columns or 1.8g on the 10.0g columns.
Load the crude mixtures using pipettor in a polar solvent (usually methanol).
Page 14

RR99830 - Carousel Work-Up Station
Guidelines for Using SPE Columns - continued
4. Elution of impurities
Impurities are normally eluted using the same solvent as is used for solvating the
column. Use the Speediflow to provide a positive pressure to increase flow rates.
Silica: Elute using a non-polar solvent e.g. hexane to remove non-polar impurities
(or non-polar products).
SCX: Elute with a polar solvent e.g. methanol to remove non-basic compounds.
If SCX is being used to remove basic impurities the non–basic compounds may
be the desired products.
SAX Acetate: Elute with a polar solvent e.g. methanol to remove non-acidic
compounds. If SAX Acetate is being used to remove acidic impurities the non–
acidic compounds may be the desired products.
Aminopropyl: May be used as a polar normal phase sorbent (treat as Silica)
or an ion-exchange column (treat as SAX Acetate). Aminopropyl is particularly
useful for removing strongly acidic impurities when used as an ion-exchanger.
5. Elution of product
Silica: Use a more polar solvent for elution. For example if hexane was used
for the solvating/equilibrating solvent hexane/ethyl acetate may be a suitable
eluting solvent. The elution solvent should be determined from thin layer
chromatography (tlc). Aim for an Rf of 0.4-0.6 to elute the desired product(s) in a
small solvent volume. A compound with an Rf of 0.5 will elute in approximately 2
column volumes (CVs). (No of CVs = 1/Rf)
SCX: Use a basic medium in a polar solvent (same solvent as used to solvate
/equilibrate column) to elute basic products. For example if the column were
solvated/equilibrated with methanol, 10% aqueous ammonia in methanol (1CV)
would be a suitable eluant.
SAX Acetate: Use an acidic medium in a polar solvent (same solvent as used to
solvate/equilibrate column) to elute acidic products. For example if the column
was solvated/equilibrated with methanol, 10% acetic acid in methanol (1CV)
would be a suitable eluant.
Aminopropyl: May be used as a polar normal phase sorbent (treat as Silica) or
an ion-exchange column (treat as SAX Acetate).
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