Care and Use Notes for Lux SFC

(Supercritical Fluid Chromatography)

Thank you for purchasing Lux columns for your chiral work. Below are recommended instructions for the care
and use of your Lux analytical column under SFC conditions (ID ≤ 4.6 mm).

General Information Column Setup and Use under SFC

Column Description Column Setup and Mobile Phase Co-solvent
Column Name Phase Description Install the column in the SFC instrument oven compartment, set SFC instrument
backpressure regulator around 80-100 bars and equilibrate the column with a
Lux Amylose-1 Amylose tris(3,5-dimethylphenylcarbamate)
minimum of ten column volumes of the SFC mobile phase prior to use. A good
Lux Cellulose-1 Cellulose tris(3,5-dimethylphenylcarbamate) starting choice for SFC mobile phase is CO2/MeOH or CO2/EtOH (80:20, v/v) with
Lux Cellulose-2 Cellulose tris(3-chloro-4-methylphenylcarbamate) or without additives. We recommend increasing flow rate gradually to prevent
backpressure from going above 300 bar (4300 psi) for analytical column dimen-
Lux Cellulose-3 Cellulose tris(4-methylbenzoate) sions. For Lux preparative columns in SFC mode, do not exceed 3500 psi back
Lux Cellulose-4 Cellulose tris(4-chloro-3-methylphenylcarbamate) pressure. Recommended flow rates for various dimensions in SFC are presented
in the table below.
Lux i-Amylose-3 Amylose tris(3-chloro-5 methylphenylcarbamate)
Lux i-Cellulose-5 Cellulose tris(3,5-dichlorophenylcarbamate) Column ID Optimal Flow Rate Max. Backpressure

Lux i-Amylose-1 Amylose tris(3,5-dimethylphenylcarbamate) 4.6 mm 3-6 mL 4300 psi

21.2 mm 65-125 mL 3500 psi
Shipping Solvent 30.0 mm 125- 250 mL 3500 psi
n-Hexane/2-propanol (9:1, v/v)
50.0 mm 350-700 mL 3500 psi
Test Certificate
Mobile Phase Additives
Each column is individually tested before shipment. A test certificate showing the
separation parameters for trans-stilbene oxide is enclosed with each column. A For some basic or acidic chiral compounds, it may be necessary to use an ap-
test certificate can also be found on our website using column part number and propriate mobile phase additive to achieve chiral resolution and to insure proper
serial number. peak shapes. Ammonia (NH4OH), diethylamine, ethanolamine, isopropyl amine
(IPA) and butyl amine in the concentration range 0.1-0.5 % can be used with
Operating Backpressure basic analytes, while trifluoroacetic acid (TFA) or acetic acid (0.1-0.5 %; typically
0.1-0.2 %) can be used with acidic analytes. Mixtures of basic and acidic mobile
The mobile phase flow rate should be set such that the column backpressure phase additives are acceptable (for example IPA with TFA). Lux columns will
stays below 300 bar (4300 psi) for analytical columns and below 241 bar (3500 deliver consistent results when operated with mobile phases containing additives
psi ) for Axia packed preparative columns. at the concentration levels specified above. However, a decrease in column
efficiency may occur when a column is used in combination with these additives.
Operating Temperatures Therefore, we advise to dedicate columns to mobile phases containing basic or
With standard co-solvent/modifier (such as alcohol and acetonitrile) the column acidic additives.
can be used in the temperature range 0-50 °C.
Screening Strategies for Successful Separation
Mobile Phase Restrictions See Phenomenex Technote TN-9003 and references cited therein.
Coated Lux chiral stationary phases are prepared by coating silica with various
polysaccharide derivatives. Therefore, any solvent dissolving the polysaccharide
derivative (such as tetrahydrofurane, acetone, chlorinated hydrocarbons, ethyl- Column Care, Maintenance, and Storage
acetate, dimethylsulfoxide, dimethylformamide, N-methylformamide, toluene,
methylethyl ketone and methyl tert-butyl ether, etc…) must be avoided even Extending Lifetime and Removal of Contaminants
in trace amounts (e.g. even as sample solvent). Co-solvent such as methanol Phenomenex recommends the use of SecurityGuard™ guard cartridges to extend
(MeOH), ethanol (EtOH), isopropanol (IPA) as well as acetonitrile are compatible the lifetime of your column. Ideally, samples must be completely dissolved in the
and typically used under SFC conditions. mobile phase modifier and filtered through a syringe filter of approximately 0.45
μm porosity. To remove potential contaminants after extended use of your Lux
Normal Phase to SFC Conversion column, we recommend flushing the column with 100 % methanol or ethanol for
Lux Columns are shipped in Normal Phase solvent and will require solvent for 2-3 hours at the appropriate flow rate. Backflush can also be used to clean
switching before running it in SFC mode. Phenomenex recommends Flushing the column.
your column with ten column volumes of MeOH: EtOH 90:10 followed by your
SFC mobile phase for 10 column volumes. Column Storage
In SFC mode, when shutting down the instrument, the column pressure should
be allowed to go to ambient gradually. Rapid depressurization of the column can
lead to column and/or system damage. Refer to your SFC instrument manual for
depressurization protocol.

Short term Storage

Ideal choice for storage will be as per long term storage. However if the column
is supposed to be used within couple of days, you may depressurize the column
to atmospheric pressure and add endplugs to both ends.
Lux and SecurityGuard are trademarks of Phenomenenex.
© 2022 Phenomenex Inc. All rights reserved. Longterm storage
FL33750822_W

Columns used in SFC mode must be flushed with 10 column volume of methanol
or ethanol or other co-solvent alcohol that you anticipate to use in SFC method.
Have questions or want more details on implementing this method? We would love to help!
Visit www.phenomenex.com/Chat to get in touch with one of our Technical Specialists