Supelco 3 Chiral PDF

Table of Contents 5

CHIRAL CHROMATOGRAPHY
Think Chiral...Think Supelco 2 Chiral GC Columns 21
Cyclodextrin-based GC CSPs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21
sigma-aldrich.com/chiral 2 Choosing a Chiral GC Column . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
Chiral GC Column Screening Kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
Astec: Part of the Sigma-Aldrich Analytical Astec CHIRALDEX® GC Column Screening Kit . . . . . . . . . . . . . . . . . . . . . . 23
Family 2 Supelco DEX™ GC Column Screening Kit . . . . . . . . . . . . . . . . . . . . . . . . . . 23
Group 1: Surface Interactions, Complex Derivatives . . . . . . . . . . . . . . . . . . 23
Chiral Services 2 Group 2: Surface/Inclusion Interactions, Simple Derivatives . . . . . . . . . . . . 26
Group 3: Inclusion Interactions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
Chiral HPLC & SFC Columns 3 Chiral GC Column Protection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
Rapid, Efficient and Effective Chiral Method Development . . . . . . . . . .. . . 3 Chiral GC Column Test Mixes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
Preparative Chiral Separations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . 3
Supercritical Fluid Chromatography (SFC) . . . . . . . . . . . . . . . . . . . . . . . .. . . 3 Chiral Derivatization Reagents 32
Simulated Moving Bed Chromatography (SMB) . . . . . . . . . . . . . . . . . . .. . . 4 ChiraSelect™ HPLC Derivatization Reagents . . . . . . . . . . . . . . . . . . . . . . . . 32
Choosing a Chiral HPLC Column . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . 4 ChiraSelect™ GC Derivatization Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . 33
Astec CHIROBIOTIC® Chiral HPLC Columns . . . . . . . . . . . . . . . . . . . . . . .. . . 4
Astec CYCLOBOND® Chiral HPLC Columns . . . . . . . . . . . . . . . . . . . . . . .. . . 9 Chiral Mobile Phase Additives 34
Astec Cellulose DMP (Dimethylphenylcarbamate) . . . . . . . . . . . . . . . . .. . 13
Astec P-CAP™ and P-CAP™-DP Chiral HPLC Columns . . . . . . . . . . . . . .. . 14
Astec CLC-L and CLC-D (Copper Ligand Exchange) . . . . . . . . . . . . . . . .. . 15
Protein-based Chiral HPLC Columns . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . 16
Kromasil® Chiral HPLC Columns . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . 18
Cyclofructans: LARIHC™ and FRULIC™ Chiral HPLC and HILIC Columns . . 19
Protection for Chiral HPLC Columns . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . 19
Chiral HPLC Column Test Mixes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . 21

For all of your analytical needs, visit us at sigma-aldrich.com/analytical

To order, visit sigma-aldrich.com/order
2 Chiral Chromatography
Think Chiral...Think Supelco
8P R O D U C T S
Astec: Part of the Sigma-Aldrich
Think Chiral...Think Supelco Analytical Family
From a separation perspective, few types of compounds can match the
challenges posed by chiral compounds. Chirality is important primarily
because biological systems recognize stereochemistry. The enantiomers of
chiral drug substances usually have different therapeutic efficacies, and it is
not uncommon for one enantiomer to have unwanted and even toxic
qualities. Eliminating the inactive enantiomer in chiral agrochemicals can A pre-eminent innovator in chiral chromatography, Astec, its products and
reduce by half or more the amount of chemical that needs to be applied to the expertise and dedication of its people became part of Sigma-Aldrich in
the crop, with less waste and less environmental impact. However, chiral 2006. Many Astec products were conceived through collaboration with Prof.
compounds pose a particular analytical challenge: Enantiomers have Daniel Armstrong under a spirit of innovation that continues to thrive in our
identical physical and chemical properties and differ only in their optical R&D group today.
rotation and interactions with other chiral molecules. Chiral Columns + Chiral Reagents + Chiral Services +
Expert Customer Support = Successful Enantiomer Separations
Astec columns are backed by our world-class customer and technical
support. They have become an integral part of our chiral offering, which
includes:
• HPLC & SFC columns - Choices in stationary phase chemistry allow
versatile mobile phase selection, suitable for a wide variety of analytes
• Capillary GC columns - Wide choice in inert, low-bleed, coated and
chemically-bonded highly-enantioselective cyclodextrin derivatives
• Reagents - Selective chiral derivatization reagents and high-purity chiral
mobile phase additives
• Chiral Services - Chiral column screening (HPLC and GC), method
development and optimization and small-scale purification
Chiral Services
Developing chiral methods and isolating pure enantiomers for further
testing can be time consuming. To help our customers, we offer chiral
column screening, method optimization and isolation of mg to gram
quantities of purified enantiomer. Process-scale amounts can be purified by
sigma-aldrich.com/chiral our SAFC facilities around the world. All work done by the expert staff of our
state-of-the-art Chiral Services Laboratory is performed according to your
Corporate web portal devoted to all things chiral specifications and is fully confidential. Our laboratory personnel also perform
Sigma-Aldrich is a leader in chiral products and services for chemical achiral separations and purifications.
synthesis, drug discovery, and analytical assessment. Our regularly-updated Chiral Services
chiral web portal presents all of our products, services, seminars and
technical literature for chirality in one convenient location. The site can be HPLC Chiral Column HPLC chiral column screening protocol includes multiple mobile
Screening phase conditions run on multiple chiral stationary phases
used to access Supelco′s chiral HPLC and GC columns, our Chiral Services representing four separation modes (NP, RP, PIM and POM).
Laboratory, as well as Aldrich® chiral chemistry products, like privileged Positive separation is verified on a separate system. Enantiomers
ligands and complexes, chiral catalysts, ligands and reagents, chiral are identified as (+) and (-)
auxiliaries and chiral building blocks. GC Chiral Column GC column screening involves exploration of 4 GC chiral phases.
Screening Samples that require derivatization are verified by GC-MS.
Your resource for technical literature, bibliography, and applications for
HPLC and GC Chiral Method optimization may vary, depending on the intended use of
chiral chromatography Method Optimi- the method, which may include isolation/purification of
Developing a new chiral method typically involves perusing application zation enantiomers, resolution of metabolites, establishment of minimum
and Development detection limits, LC-MS compatible methods for clinical, stability or
databases, consulting the literature, screening columns, or contracting with Services dissolution studies. Typical experiments in the optimization study
a chiral service lab. Supelco can help with all of these approaches. We have include modifying buffer and pH, organic modifier type and
created a centralized location of chiral resources and facile navigation. Visit strength, and column temperature.
and bookmark our chiral web portal to gain access to the valuable chiral Small-Scale Enantio- Milligram to gram quantities. Typical enantiomeric purity is 98%
resources: meric Purification and verification is determined by analytical methods established in
the screening study. Larger scale purifications are available
• Applications: A growing list of thousands of applications on Astec and through our SAFC offices worldwide.
Supelco columns
• Bibliography: Up-to-date compilation of citations using our chiral columns
• Technical literature, handbooks and presentations
sigma-aldrich.com/analytical View our current seminar and tradeshow schedule at sigma-aldrich.com/analytical-events

Chiral Chromatography 3
Chiral Services
Method Development and Column Screening Protocol on CHIRO-

BIOTIC® and CYCLOBOND HPLC Columns
CHIROBIOTIC® V2, CYCLOBOND I 2000,
T, R, TAG HP-RSP, DMP, DNP
Mobile
Phase Screening Screening
System Mobile Phase Mobile Phase Parameters to Optimize
Polar Ionic (100:0.1:0.1, v/v/v) Change acid-base ratio,
CH3OH/acetic acid/ change the type of acid
triethylamine or base, add a volatile salt
(test different ammonium
salts)
Reversed- (30:70) CH3CN/20 mM (1) (30:70) CH3CN/20 mM Change the % and type
Phase ammonium acetate, ammonium acetate, pH 5 of organic modifier,
pH 5 (2) (20:80) CH3OH/20 mM adjust pH, buffer type and
ammonium acetate, pH 5 ionic strength
Polar Methanol 95:5:0.1:0.1; CH3CN/ Use other polar organic
Organic CH3OH/acetic acid/ solvents or blends
triethylamine
Normal (30:70) Ethanol/ (30:70) Ethanol/heptane Increase % of polar
Supelco Chiral Services Laboratory Phase heptane (DMP, DNP only) modifier, change both
solvents
Chiral HPLC & SFC Columns The Astec CHIROBIOTIC® and CYCLOBOND CSPs we recommend in the
Supelco′s HPLC chiral stationary phases (CSPs) cover a broad range of screening protocol are available in convenient kits. Also, you can increase
chemistries, enantioselectivity, and application focus. The unique, propri- the probability of success by incorporating the Astec Cellulose, Astec P-CAP,
etary Astec CHIROBIOTIC® and Astec CYCLOBOND® are particularly Astec CLC, LARIHC, and protein-based CSPs into your screening protocol.
interesting from a chiral method development standpoint. Currently, the We would be happy to help you select the best line-up of columns for your
chiral HPLC and SFC columns we carry include: types of analytes, detectors, and preferred mobile phase systems.
• Astec CHIROBIOTIC® macrocyclic glycopeptide-based CSPs

• Astec CYCLOBOND® bonded cyclodextrin-based CSPs Related Information
• Astec Cellulose DMP polysaccharide-based CSPs
Request free literature by phone, fax, or visit sigma-aldrich.com/literature.
• Astec P-CAP™ and P-CAP™-DP chiral polymer-based CSPs
• Astec CLC copper ligand exchange CSPs No. Title
• CHIRALPAK® AGP, CBH, and HSA protein-based CSPs from DAICEL Corp. T409107 Chiral Method Development Wallchart
• LARIHC™ and FRULIC™ cyclofructan-based CSPs from AZYP, LLC
• Kromasil® AmyCoat®, CelluCoat®, Chiral DMB, and Chiral TBB from Eka
Chemicals AB (available from Sigma-Aldrich in USA, Canada, and
Puerto Rico) Preparative Chiral Separations
The development of innovative, new CSPs is an important R&D activity It is often the case that mg to gram quantities of purified enantiomer are
for us. Please call our Technical Services or visit our web site, www. required for safety and efficacy testing, or for further modification. Chiral
sigma-aldrich.com/chiral, for information on our most current offering. HPLC is commonly used for this application. Once an enantiomer is shown
to have desirable characteristics, then an asymmetrical synthesis may be
developed to avoid a racemate. However, if a cost-effective synthetic
Rapid, Efficient and Effective Chiral Method approach is not readily available, then chromatography may indeed provide
the best means to obtain purified enantiomer. Supelco′s HPLC CSPs are
Development amenable to preparative separations, whether in classic LC mode, SFC or in
Successful separations are more likely when you include Astec continuous preparative techniques such as SMB and multi-column
CHIROBIOTIC® and Astec CYCLOBOND columns in your chiral column processes. Supelco chiral HPLC phases offer benefits in:
screening protocol along with conventional cellulosic/amylosic CSPs. These • Sample Solubility - Choose mobile phases that maximize sample
two types of CSPs are highly complementary. For developing a new chiral solubility, including aqueous and polar systems
HPLC method, we have created and use routinely in our laboratories a • Throughput - Shorter retention times give higher throughput
simple and rapid chiral column screening protocol shown below. Method
development follows a simple strategy that tests polar ionic, polar organic,
reversed-phase and normal phase modes. Supercritical Fluid Chromatography (SFC)
SFC is gaining in popularity primarily because of its speed and "green"
advantages over normal phase HPLC. The CO2 is readily removed from the
eluate, which makes it ideal for prep. We offer several types of Astec chiral
HPLC columns that are suitable for SFC separations.
• Astec Cellulose DMP columns are the most ideally suited for chiral SFC.
They provide rapid separations with excellent enantioselectivity, long
column lifetime, and low backpressure without significant column bleed.
• Astec P-CAP columns have recently been found to be beneficial in the
separation of a complex mixture of enantiomers and achiral impurities.
They provided resolution in instances where conventional polysaccharide-
based CSPs failed (1).
For hazardous product information, visit sigma-aldrich.com/safetycenter

Chiral HPLC & SFC Columns

Supercritical Fluid Chromatography (SFC)
• A number of the Astec CYCLOBOND derivatives offer good opportunities

for SFC, including CYCLOBOND I 2000 DNP, a pi acid, and the
Astec CHIROBIOTIC® Chiral HPLC Columns
CYCLOBOND I 2000 DMP, a pi base. Astec CYCLOBOND I 2000, RSP and
DM are also useful for SFC, the DM especially for fused polycyclic
compounds. Since these latter CSPs have as their primary mechanism Related Information
steric repulsion and hydrogen bonding, SFC's benefits of speed and Request free literature by phone, fax, or visit sigma-aldrich.com/literature.
efficiency are realized. No. Title
• Astec CHIROBIOTIC® CSPs are suitable for polar and non-polar neutral T408131 CHIROBIOTIC® Brochure
analytes. However, because of their ionic character, additives are required
for ionized analytes to avoid lengthy analysis times. A good summary
appears in Liu, et al (2).
CHIROBIOTIC®: "Chiral by Nature"
(1) Barnhart, W. W.; Gahm, K. H.; Hua Z.; Goetzinger, W. Supercritical Fluid
Chromatography Comparison of the Poly(trans-1,2-Cyclohexanediyl-bis The Astec CHIROBIOTIC®; family comprises highly enantioselective chiral
Acrylamide) (P-CAP) Column with Several Derivatized Polysaccharide-based HPLC stationary phases based on naturally-occurring macrocyclic glyco-
Stationary Phases. J. Chromatogr. B, 2008, 875, 217-229. peptides that have been bonded through multiple covalent linkages to high
(2) Liu, Y.: Berthod, A.; Mitchell, C. R.; Xiao, T. L.; Zhang, B.; Armstrong, D. W. purity silica particles. Developed by Prof. Daniel Armstrong (1), CHIRO-
Super/Subcritical Fluid Chromatography Chiral Separations with Macrocyclic BIOTIC®; CSPs are unique in possessing ionic functional groups, which
Glycopeptide Stationary Phases. J. Chrom. A, 2002, 978, 185-204. means they can be used for reversed-phase and LC-MS separation of
ionizable enantiomers, as well as neutral molecules. The members of the
CHIROBIOTIC®; family have complementary stereoselectivity. If one CHIRO-
Simulated Moving Bed Chromatography BIOTIC®; CSP does not give baseline resolution, testing the other CHIRO-
BIOTIC®; CSPs in the same mobile phase often results in complete resolution.
(SMB)
Astec CHIROBIOTIC®; features and application areas:
A continuous preparative HPLC technique, SMB or counter-current
• Aqueous and non-aqueous separations on the same column
chromatography can be conceptualized as multiple columns used in series
• Ideal for reversed-phase and polar mobile phases for LC-MS
to make a single column of effectively infinite length. Supelco chiral HPLC
• No solvent or additive memory effects
columns and packings permit robust SMB operation. The polar organic and
• Robust columns with long lifetimes, important in bioanalysis
polar ionic (methanol or acetonitrile containing soluble ionic additives)
• Solvent choices maximize sample solubility
mobile phases and larger particle sizes of Astec CHIROBIOTIC®; and Astec
• Excellent preparative scalability and capacity
CYCLOBOND CSPs provide minimal back pressure, which is important in
• Fast kinetics for speed and efficiency
SMB to maximize through-put by allowing high flow rates. Additionally,
mobile phases can be chosen to maximize sample solubility to prevent (1) Armstrong, D. W.; Tang, Y.; Chen, S.; Zhou, Y.; Bagwill, C.; Chen, J.
precipitation and increase the sample load. Note the special section on SMB Macrocyclic Antibiotics as a New Class of Chiral Selectors for Liquid
sets in the Astec CHIROBIOTIC®; products. Chromatography. Anal. Chem. 1994, 66, 1473-1484.
CHIROBIOTIC® CSPs—Physical Properties

Related Information Chiral Chiral Sugar Inclusion pH
Request free literature by phone, fax, or visit sigma-aldrich.com/literature. CHIROBIOTIC® CSP Selector Centers Groups Cavities Range
CHIROBIOTIC® V and V2 Vancomycin 18 2 3 3.5–7.0
No. Title
T409105 Astec CHIROBIOTIC® Columns Sets for SMB CHIROBIOTIC® T and T2 Teicoplanin 23 3 4 3.8–6.8
CHIROBIOTIC® TAG Teicoplanin 8 0 4 3.0–6.8
aglycone
CHIROBIOTIC® R Ristocetin A 38 6 4 3.5–6.8
Choosing a Chiral HPLC Column
hydrogen bonding &
When performing a chiral separation, it is usually very difficult to predict dipole stacking sites
which CSP will provide adequate enantioselectivity, especially when
working with new chemical entities. Even the experts use a column ionic
screening protocol. To make this process easier for you, we offer three
economical and time-saving options:
1. Access our extensive applications and bibliographical database by calling C ionic
our Technical Services or viewing the growing applications library on our
web site. A B
π-acceptor
2. Purchase an Astec CHIROBIOTIC® and/or Astec CYCLOBOND column
screening kit. The kits contain CSPs in column geometries that have the
highest success rate. They are priced below what the columns would cost if
purchased separately. A, B, C are inclusion sites
3. Rely on the expertise and professionalism of our Chiral Services laboratory sugar
molecules
for column screening, method optimization, and small-scale purification.
Proposed structure of vancomycin (the chiral selector in CHIROBIOTIC® V and V2)

showing different types of molecular interactions. The presence of ionic interactions is
what differentiates CHIROBIOTIC® CSPs from other CSPs, and makes them
valuable for polar and ionic compounds and MS detection.
sigma-aldrich.com/analytical Access our analytical literature and newsletters at sigma-aldrich.com/literature


Astec CHIROBIOTIC® Chiral HPLC Columns: Unique Multi-Modal Operation Includes Ionic Interactions
Unique Multi-Modal Operation Includes Ionic The Most Versatile HPLC CSP
Interactions Astec CHIROBIOTIC® CSPs offer the flexibility in choice of mobile phase
conditions, both aqueous and non-aqueous, and are ideal for analytical and
Astec CHIROBIOTIC® CSPs offer six different types of molecular interactions
preparative separations of neutral, polar and ionic compounds. Their
on one column: ionic, H-bond, pi-pi, dipole, hydrophobic and steric. They
multiple interactions and absence of memory effects means the same
also possess multiple inclusion cavities that influence selectivity based on
CHIROBIOTIC® column can be successfully used in a variety of mobile
the molecular shape of the analyte. The optimization of enantiomer
phases, a significant benefit over CSPs that operate only in a single mode,
resolution is achieved by changing the mobile phase to leverage the types
normal or reversed-phase, for example, and must be dedicated to those
and relative strengths of the various interactions.
mobile phase systems.
HPLC Analysis of Beta-Receptor Agonist Enantiomers on Astec®
Normal Phase Mode Polar Ionic Mode
CHIROBIOTIC® T (n-Hexane / Ethanol) (Method / Acetic Acid / TEA)
 application for HPLC

8.31 min.
column .............................................. Astec CHIROBIOTIC® T, 25 cm x 4.6 mm, 5 μm particles (12024AST) 25.8 min.
mobile phase ............................................................................................... 15 mM ammonium formate in methanol
flow rate ...................................................................................................................................................................................... 1 mL/min t-Bu t-Bu 27.5 min.
column temp. .................................................................................................................................................................................... 25 °C t-Bu
detector ............................................................................................................................................................................. UV at 220 nm N
P
Application No. .................................................................................................................................................................... G004337 H3CO
t-Bu 9.12 min. O
t-Bu
F
H3CO
P
t-Bu H3C N HBr
t-Bu t-Bu CH3
Application courtesy of Dr. M. Althous,

Hoffman la Roche, Basel, Switzerland
Demonstration of CHIROBIOTIC® column utility in both normal phase

and polar ionic modes. The same CHIROBIOTIC® column can be used in all four
modes, from aqueous to organic, without memory effects or loss of performance.
Column: CHIROBIOTIC® V, 25 cm x 4.6 mm, 5 μm particles (11024AST).
Left: Normal phase mode. 3,5-tBu-MeOBIPHEP enantiomers.
n-Hexane:10% ethanol in n-hexane (75:25), 1.5 mL/min.
Right: Polar ionic mode. Citalopram enantiomers.
Methanol:acetic acid:TEA (99.8:0.1:0.1), 0.5 mL/min.
Astec CHIROBIOTIC® Application Areas

Astec CHIROBIOTIC® CSPs have found utility in many areas of analytical
chemistry, including:
• Drug Discovery - High enantioselectivity, fast screening protocols,
scalability to prep, reproducibility for reliable methods, polar and non-
polar analytes
• Bioanalytical, Drug Metabolism - High throughput, MS-compatibility,
aqueous samples, short run times, rugged columns
• Amino Acid and Peptide Analysis - Resolves underivatized natural and
synthetic chiral amino acids and peptides; different selectivity and higher
preparative capacity for achiral amino acids than C18
• Organic Synthesis - Compatible with all HPLC solvents, including
chlorinated solvents, to optimize sample solubility, fully scalable to prep
Simplified Chiral Method Development

Astec CHIROBIOTIC®; HPLC columns enable simple method development,
and are particularly useful for polar compounds due to the unique polar
ionic mode. A single CHIROBIOTIC®; column possesses multiple types of
molecular interactions and can be run in four distinct modes. The same
column can be exposed to all of the conditions outlined in the screening
protocol without any change or loss of performance. This versatility is just
one advantage that CHIROBIOTIC®; CSPs have over other CSPs.
View our technical presentations at sigma-aldrich.com/analytical-videos


Astec CHIROBIOTIC® Chiral HPLC Columns: Complementary Selectivity to Cellulosic/Amylosic CSPs, but with Benefits
Complementary Selectivity to Cellulosic/Amylosic HPLC Analysis of Clenbuterol Enantiomers on Astec® CHIRO-

BIOTIC® T (ESI-MS of Plasma Extract)
CSPs, but with Benefits
 application for HPLC
Astec CHIROBIOTIC®; CSPs will perform the desired separation in nearly 75% column ..................................... Astec CHIROBIOTIC® T, 10 cm x 2.1 mm I.D., 5 μm particles (12018AST)
of the cases, with a 50% overlap of the cellulosic/amylosic phases. However, mobile phase ............................................................................................... 10 mM ammonium formate in methanol
the CHIROBIOTIC®; CSPs often provide significant advantages, like allowing flow rate .................................................................................................................................................................................. 0.3 mL/min
mobile phases that are better suited to the sample and detection method, column temp. .................................................................................................................................................................................... 30 °C
detector .................................................................................................................................................................................................. ESI(+)
or the CHIROBIOTIC®; method may be faster, more efficient or more robust. sample ............... clenbuterol, 10 ng/mL in rat plasma (phospholipids removed by extraction with
A CHIROBIOTIC®; method may also have advantages from a preparative HybridSPE-Phospholipid)
standpoint in terms of solvent selection and capacity. Application No. .................................................................................................................................................................... G004245
Cellulosic/ OH
CHIROBIOTIC® Overlap Clenbuterol
amylosic 700
(~25%) (~50%) CI
(~25%)
600 CH3
When performing chiral HPLC column screening, most enantiomers can be resolved on both HN
CHIROBIOTIC® and cellulosic/amylosic CSPs. However, CHIROBIOTIC® CSPs often work better with H2N
500 CH3
ionic and highly polar compounds. Even in the areas of overlap, CHIROBIOTIC® CSPs often have H3 C
CI
advantages in solubility, LC/MS-compatibility and throughput.
400
cps
LC-MS-friendly Chiral Separations 300
Each MS ionization source has an optimal set of mobile phase conditions. Clenbuterol
Outside this set, ionization may be suppressed with resulting loss in 200
sensitivity. Astec CHIROBIOTIC®; phases are uniquely able to operate across Phospholipids
all mobile phase systems. CSPs that are limited to normal phase operation, 100
like the majority of cellulose-based CSPs, reduce the analyst′s options in
detection methods. Astec CHIROBIOTIC®; columns can be used in 0
0 1 2 3 4 5
conjunction with HybridSPE-Phospholipid plates to enhance sensitivity by Min
completely removing endogenous proteins and phospholipids, as shown in
the clenbuterol from rat plasma application that follows.
HPLC Analysis of Ketoprofen Enantiomers on Astec® CHIRO- Ideally Suited for Preparative Applications
BIOTIC® R (MS Detection) Astec CHIROBIOTIC® phases offer unique opportunities for preparative
 application for HPLC purifications.
column .............................................. Astec CHIROBIOTIC® R, 15 cm x 2.1 mm, 5 μm particles (13019AST) • Excellent economics - Especially with the polar organic and polar ionic
mobile phase ............................. A: 20 mM ammonium acetate, pH 5.6 B: methanol Ratio: 70:30 (A:B)
flow rate .................................................................................................................................................................................. 0.2 mL/min
modes. Ionic interactions play a significant role in the chiral recognition
column temp. .................................................................................................................................................................................... 35 °C mechanism on Astec CHIROBIOTIC® CSPs. Solvents here are anhydrous
detector ................................................................................................................................................................................................... ESI(-) and more volatile and less toxic than the typical normal phase mode.
sample ........................................................................................................................................................................................ ketoprofen • No solvent limitations - Halogenated solvents and very polar solvents are
Application No. .................................................................................................................................................................... G004331 well tolerated on Astec CHIROBIOTIC® CSPs. This solvent tolerance is
especially useful when optimizing for sample solubility.
• Versatility - The same Astec CHIROBIOTIC® column can be run in four
3.46
distinctly different mobile phase types. Use of acid/base does not
4.31 preclude their use in other mobile phases.
O CH3
• Stability - Exceptional long-term stability of Astec CHIROBIOTIC® CSPs is
36 O derived from the multiple linkages used in anchoring the CSP and to the
mild run conditions that are typically required.
OH • Capacity - The range of capacities is compound dependent. Significantly
overlaps cellulose and amylose phases based on throughput, primarily
Response
24 because separations on Astec CHIROBIOTIC® CSPs are usually very fast.

Capacities on Astec CHIROBIOTIC® V2/T2 phases are ~2.5 mg/gm (α = 1.5).
Maximum capacity achieved was ~300 mg on column using a 250 x 21.2
mm column with α = 2.0.
12
0
0.98 1.00
0 5 10
Min
sigma-aldrich.com/analytical To order, visit sigma-aldrich.com/order


Astec CHIROBIOTIC® Chiral HPLC Columns: Astec CHIROBIOTIC® Column Screening Kits
Astec CHIROBIOTIC® Column Screening Kits Astec CHIROBIOTIC® V Chiral HPLC Guard
The four Astec CHIROBIOTIC®; CSPs we recommend in the screening I.D. (mm) L (cm) Cat. No. Qty
protocol are available in 25 cm or 10 cm column kits. A full description of particle size 5 μm
the screening procedure and instructions on how to optimize the 1.0 2 11101AST 1 ea
separation are included with each kit. 4.0 2 11100AST 1 ea
Kit components:
The 4 mm I.D. guard cartridge requires a holder, Cat. No. 21150AST (stand-
• Astec CHIROBIOTIC®; T2 alone) or 504254 (direct-connect), sold separately. The 1 mm I.D. guard is
• Astec CHIROBIOTIC®; V sold complete and does not require a separate holder.
• Astec CHIROBIOTIC®; R
• Astec CHIROBIOTIC®; TAG Astec CHIROBIOTIC® V2 Chiral HPLC Column
• Astec CHIROBIOTIC®; Handbook
I.D. (mm) L (cm) Cat. No. Qty
You can further expand the screening field by incorporating Astec Cellulose particle size 5 μm
DMP, Astec CYCLOBOND, Astec P-CAP, Astec CLC, LARIHC, and protein- 2.1 10 15018AST 1 ea
based CSPs (sold separately) into your screening protocol. 2.1 15 15019AST 1 ea
2.1 25 15020AST 1 ea
Astec CHIROBIOTIC® HPLC Column Screening Kit
4.6 5 15021AST 1 ea
Description Cat. No. Qty 4.6 10 15022AST 1 ea
Astec CHIROBIOTIC® HPLC Column 10300AST 1 kit 4.6 15 15023AST 1 ea
Screening Kit, particle size 5 μm, L 10 cm ×
4.6 25 15024AST 1 ea
I.D. 4.6 mm
10.0 25 15034AST 1 ea
Astec CHIROBIOTIC® HPLC Column 10305AST 1 kit
Screening Kit, particle size 5 μm, L 25 cm × 21.2 25 15044AST 1 ea
I.D. 4.6 mm 30.0 25 15054AST
particle size 10 μm
4.6 25 15124AST 1 ea
Astec CHIROBIOTIC® V and V2 (Vancomycin) particle size 15 μm
4.6 25 51041AST 1 ea
Neutral molecules, amides, acids, esters and amines show considerable
enantioselectivity on these vancomycin-based CSPs. A wide variety of
secondary and tertiary amines have been separated on the Astec Astec CHIROBIOTIC® V2 Chiral HPLC Guard
CHIROBIOTIC® V in the polar ionic mode. Astec CHIROBIOTIC® V has I.D. (mm) L (cm) Cat. No. Qty
demonstrated many of the separation characteristics of protein-based particle size 5 μm
stationary phases, but with exceptional stability and much higher sample
1.0 2 15101AST 1 ea
capacity. Some chiral analytes have been resolved that have not been
4.0 2 15100AST 1 ea
reported separated on any other chiral stationary phase. Astec CHIRO-
BIOTIC® V and V2 differ in their bonding chemistry the pore size of the
support particle, giving them different selectivity and preparative capacity.
alone) or 504254 (direct-connect), sold separately. The 1 mm I.D. guard is
• Bonded phase: Vancomycin sold complete and does not require a separate holder.
• Operating pH range: 3.5 - 7.0
• Particle type: High-purity, spherical silica
• Particle diameter: 5 or 10 μm (other particles sizes, please inquire) Astec CHIROBIOTIC® T and T2 (Teicoplanin)
• Pore size: 100 Å (CHIROBIOTIC® V) or 200 Å (CHIROBIOTIC® V2) Astec CHIROBIOTIC® T and T2 have teicoplanin as the chiral selector. They
For other column dimensions, particle sizes and bulk material, please offer unique selectivity for a number of classes of molecules, specifically
inquire. underivatized α, β, γ and cyclic amino acids, N-derivatized amino acids,
hydroxy-carboxylic acids, acidic compounds including carboxylic acids and
Astec CHIROBIOTIC® V Chiral HPLC Column phenols, small peptides, neutral aromatic analytes and cyclic aromatic and
aliphatic amines. Separations normally obtained on a chiral crown ether or
I.D. (mm) L (cm) Cat. No. Qty ligand exchange-type CSPs are also possible on Astec CHIROBIOTIC® T and
particle size 5 μm T2, but with much simpler mobile phases, such as alcohol-water. In addition,
2.1 10 11018AST 1 ea all of the known beta-blockers (amino alcohols), and dihydrocoumarins have
2.1 15 11019AST 1 ea been resolved. Astec CHIROBIOTIC® T and T2 differ in their bonding
2.1 25 11020AST 1 ea chemistry and the pore size of the support particle, giving them different
3.0 10 11010AST 1 ea selectivity and preparative capacity.
4.6 5 11021AST 1 ea • Bonded phase: Teicoplanin
4.6 10 11022AST 1 ea • Operating pH range: 3.8 - 6.8
4.6 15 11023AST 1 ea • Particle type: High-purity, spherical silica
4.6 25 11024AST 1 ea • Particle diameter: 5 or 10 μm (other particles sizes, please inquire)
• Pore size: 100 Å (CHIROBIOTIC® T) or 200 Å (CHIROBIOTIC® T2)
10.0 25 11034AST 1 ea
• USP Code L63
21.2 25 11044AST 1 ea
30.0 25 11054AST For other column dimensions, particle sizes and bulk material, please
inquire.


Astec CHIROBIOTIC® Chiral HPLC Columns: Astec CHIROBIOTIC® T and T2 (Teicoplanin)
Astec CHIROBIOTIC® T Chiral HPLC Column Astec CHIROBIOTIC® R (Ristocetin A)

I.D. (mm) L (cm) Cat. No. Qty CHIROBIOTIC®; R, based on the ristocetin A glycopeptide covalently bonded
particle size 5 μm to high purity silica particles, has shown particular applicability to
2.1 10 12018AST 1 ea enantiomers of acidic compounds. Selectivity on CHIROBIOTIC®; R strongly
2.1 15 12019AST 1 ea correlates to the organic modifier, favoring the alcohol-type mobile phases
2.1 25 12020AST 1 ea by a large margin.
3.0 10 12010AST 1 ea • Bonded phase: Ristocetin A
4.6 15 12023AST 1 ea • Particle diameter: 5 or 10 μm (other particles sizes, please inquire)
4.6 25 12024AST 1 ea • Pore size: 100 Å
10.0 25 12034AST 1 ea For other column dimensions, particle sizes and bulk material, please
21.2 25 12044AST 1 ea inquire.
30.0 25 12054AST
particle size 10 μm Astec CHIROBIOTIC® R Chiral HPLC Column
4.6 25 12124AST 1 ea
particle size 5 μm
10 5 51046AST 1 ea
2.1 10 13018AST 1 ea
4.6 25 51047AST 1 ea
2.1 15 13019AST 1 ea
2.1 25 13020AST 1 ea
Astec CHIROBIOTIC® T Chiral HPLC Guard
3.0 10 13010AST 1 ea
suitable for L63 per USP 4.6 5 13021AST 1 ea
I.D. (mm) L (cm) Cat. No. Qty 4.6 10 13022AST 1 ea
particle size 5 μm 4.6 15 13023AST 1 ea
1.0 2 12101AST 1 ea 4.6 25 13024AST 1 ea
4.0 2 12100AST 1 ea 10.0 25 13034AST 1 ea
21.2 25 13044AST 1 ea
The 4 mm I.D. guard cartridge requires a holder, Cat. No. 21150AST (stand- 30.0 25 13054AST
alone) or 504254 (direct-connect), sold separately. The 1 mm I.D. guard is particle size 10 μm
sold complete and does not require a separate holder. 4.6 25 13124AST 1 ea
Astec CHIROBIOTIC® T2 Chiral HPLC Column

Astec CHIROBIOTIC® R Chiral HPLC Guard
particle size 5 μm
particle size 5 μm
2.1 10 16018AST 1 ea
1.0 2 13101AST 1 ea
2.1 15 16019AST 1 ea
4.0 2 13100AST 1 ea
2.1 25 16020AST 1 ea
4.6 5 16021AST 1 ea The 4 mm I.D. guard cartridge requires a holder, Cat. No. 21150AST (stand-
4.6 10 16022AST 1 ea alone) or 504254 (direct-connect), sold separately. The 1 mm I.D. guard is
4.6 15 16023AST 1 ea sold complete and does not require a separate holder.
4.6 25 16024AST 1 ea
21.2 25 16044AST 1 ea Astec CHIROBIOTIC® TAG (Teicoplanin Aglycone)
30.0 25 16054AST
particle size 10 μm The removal of three carbohydrate moieties gives CHIROBIOTIC®; TAG
4.6 25 16124AST 1 ea complementary selectivity to CHIROBIOTIC®; T. Resolution is enhanced
toward many of the amino acids, α, β, γ and cyclic, and especially sulfur-
Astec CHIROBIOTIC® T2 Chiral HPLC Guard containing methionine, histidine and cysteine. A number of neutral
molecules like the oxazolidinones, hydantoins and diazepines, have shown
suitable for L63 per USP enhanced resolution and, more remarkably, in single-solvent mobile phases,
I.D. (mm) L (cm) Cat. No. Qty like methanol, ethanol or acetonitrile. Some acidic molecules have also
particle size 5 μm shown increased selectivity.
1.0 2 16101AST 1 ea • Bonded phase: Teicoplanin aglycone
The 4 mm I.D. guard cartridge requires a holder, Cat. No. 21150AST (stand- • Particle diameter: 5 or 10 μm (other particles sizes, please inquire)
alone) or 504254 (direct-connect), sold separately. The 1 mm I.D. guard is • Pore size: 100 Å
sold complete and does not require a separate holder.
For other column dimensions, particle sizes and bulk material, please
inquire.
sigma-aldrich.com/analytical For all of your analytical needs, visit us at sigma-aldrich.com/analytical


Astec CHIROBIOTIC® Chiral HPLC Columns: Astec CHIROBIOTIC® TAG (Teicoplanin Aglycone)
Astec CHIROBIOTIC® TAG Chiral HPLC Column

Related Information
suitable for L63 per USP
particle size 5 μm No. Title
T409105 Astec CHIROBIOTIC® Columns Sets for SMB
2.1 10 14018AST 1 ea
2.1 15 14019AST 1 ea
2.1 25 14020AST 1 ea Particle Size
(μm) L × I.D. Cat. No. Qty
4.6 5 14021AST 1 ea
phase Astec CHIROBIOTIC® V2
4.6 10 14022AST 1 ea
15 5 cm × 10 mm 51039AST 1 set
4.6 15 14023AST 1 ea
phase Astec CHIROBIOTIC® V
4.6 25 14024AST 1 ea
15 5 cm × 10 mm 51045AST 1 set
10.0 15 14232AST 1 ea
10.0 25 14034AST 1 ea
21.2 25 14044AST 1 ea
30.0 25 14054AST Astec CYCLOBOND® Chiral HPLC Columns
4.6 25 14124AST 1 ea
Related Information
Astec CHIROBIOTIC® TAG Chiral HPLC Guard Request free literature by phone, fax, or visit sigma-aldrich.com/literature.
I.D. (mm) L (cm) Cat. No. Qty No. Title

T410091 Astec CYCLOBOND® Brochure
particle size 5 μm
1.0 2 14101AST 1 ea
4.0 2 14100AST 1 ea
Bonded Cyclodextrin Stationary Phases for Chiral
The 4 mm I.D. guard cartridge requires a holder, Cat. No. 21150AST (stand- HPLC Separations
sold complete and does not require a separate holder. Cyclodextrins are produced by the partial degradation of starch and
enzymatic coupling of cleaved glucose units into crystalline, homogeneous
toroidal structures of different molecular size. Three of the most widely
Astec CHIROBIOTIC® 8-Column Sets for SMB characterized are α (alpha), β (beta) and γ (gamma) cyclodextrin. They
contain 6, 7 and 8 glucose units, respectively. Since glucose is chiral,
8P R O D U C T S cyclodextrins are chiral. For example, β-cyclodextrin has 35 stereogenic
centers. The toroidal structure has a hydrophilic surface resulting from the 2,
SMB (simulated moving bed) is a form of preparative chromatography that 3 and 6 position hydroxyl groups, which makes them water soluble. The
utilizes multiple columns that act in concert as a single column. Throughput cavity is composed of the glucoside oxygens and methylene hydrogens
using SMB can be significantly higher than batch or column format. The giving it an apolar character. As a consequence, cyclodextrins can include
qualities of Astec CHIROBIOTIC® CSPs that make them ideal for prep, the apolar portion of molecules of appropriate dimensions and bind them
including SMB, are excellent enantioselectivity, especially for polar and ionic through dipole-dipole interactions, hydrogen bonding, or London
compounds, mobile phase flexibility to maximize sample solubility, dispersion forces. Therefore, the cyclodextrin structure offers unique
versatility for operation in all mobile phases without memory effects and opportunities to separate a wide variety of isomers that have the same
high column efficiency for high throughput and minimal downstream chemical formula but differ in the spatial arrangement of their substituents.
processing. Especially relevant for prep by SMB, the ruggedness of Asetc R
R R
R
CHIROBIOTIC® CSPs enables long-term and reliable operation. The new
Astec CHIROBIOTIC® columns for SMB feature particle size and column
R R
dimensions chosen for high flow rate and high efficiency. The 8 columns in
the set have efficiencies that are matched to within 6% rsd. Single columns
in 25 cm x 4.6 mm I.D. and 5 cm x 10 mm I.D. dimensions are available for
method development and scale-up experiments. These sets are ideal for the
Octave Chromatography System manufactured by Semba Biosciences.
R R
Silica Gel
Representation of a cyclodextrin toroid molecule attached to a silica surface. The functionalized

glucose hydroxyl groups (shown as R groups in the figure) provide different enantioselectivity.
Cyclodextrins—Physical Properties
Glucose Stereogenic
Cyclodextrin Units Centers MW Cavity (nm)
Photograph of the 8-column SMB set. Each perfectly-matched column is 5 cm x 10 mm I.D. Alpha 6 30 972 0.57
Beta 7 35 1135 0.78
Gamma 8 40 1297 0.95
For technical assistance, visit sigma-aldrich.com/techinfo


Astec CYCLOBOND® Chiral HPLC Columns: Bonded Cyclodextrin Stationary Phases for Chiral HPLC Separations
CYCLOBOND is the name given to the Astec technology for bonding

cyclodextrins to high purity silica gel through a stable ether linkage. Related Information
Developed in conjunction with Prof. Daniel Armstrong (1) and introduced in Need help choosing the right chiral HPLC or GC column?
1983, this patented line of chiral stationary phases retains its ability to form Let our Chiral Services group do the work for you.
inclusion complexes, and allows for numerous chemical separations by
selectively including into the cyclodextrin cavity a wide variety of organic
molecules.
Astec CYCLOBOND® I 2000 (β-Cyclodextrin)
Astec CYCLOBOND features and application areas:
Astec CYCLOBOND I 2000 is β-cyclodextrin bonded to high purity silica by a
• Native and derivatized β- and γ-cyclodextrins
patented process to produce a stable matrix with the cyclodextrin arranged
• Covalent bonding for greater phase stability, especially in aqueous
in such a way as to retain its most valuable property of forming inclusion
systems
complexes. This allows the cyclodextrin toroids to effect numerous chemical
• High degree of selectivity from inclusion mechanism and the unusual
separations by selectively including into their cavities a wide variety of
hydrogen bonding effects of the hydrophilic surface
organic molecules. Non-inclusion type separations are also possible with the
• Additional interactions introduced by replacing some of the secondary
polar organic mode for a wide variety of molecule types.
hydroxyl groups with different selectors
• Bonded phase: Underivatized, native β-cyclodextrin
Astec CYCLOBOND I 2000 Series: • Operating pH range: 3 - 7
Based on the original CYCLOBOND I (β-cyclodextrin) technology, • Particle type: High-purity, spherical silica
CYCLOBOND I 2000 columns are second-generation products. The • Particle diameter: 5 or 10 μm
CYCLOBOND I 2000 line includes native β-cyclodextrin and eight β- • Pore size: 100 Å
cyclodextrin derivatives. • USP Code L45
Astec CYCLOBOND II Series:
For other column dimensions not listed, please inquire.
CYCLOBOND II columns are excellent chiral selectors for multi-ring
structures such as those based on anthracene, chrysene or pyrene. These are Astec CYCLOBOND® I 2000 Chiral HPLC Column
γ-cyclodextrin bonded phases, and consist of eight glucopyranose units
arranged in the same truncated cone shape. I.D. (mm) L (cm) Cat. No. Qty
particle size 5 μm
Astec CYCLOBOND Derivatives:
2.1 10 20018AST 1 ea
• Underivatized: CYCLOBOND I 2000, CYCLOBOND II
2.1 15 20019AST 1 ea
• Acetylated: CYCLOBOND I 2000 AC, CYCLOBOND II AC
2.1 25 20020AST 1 ea
• 2,3-di-O-Methyl: CYCLOBOND I 2000 DM
• 3,5-Dimethylphenyl carbamate: CYCLOBOND I 2000 DMP 4.6 5 20021AST 1 ea
• 2,6-Dinitro-4-trifluoromethyl phenyl ether: CYCLOBOND I 2000 DNP 4.6 10 20022AST 1 ea
• Hydroxypropyl ether (high performance): CYCLOBOND I 2000 HP-RSP 4.6 15 20023AST 1 ea
• Hydroxypropyl ether: CYCLOBOND I 2000 RSP, CYCLOBOND I 2000 SP 4.6 25 20024AST 1 ea
10.0 25 20034AST 1 ea
(1) Armstrong, D. W.; DeMond, W. Cyclodextrin bonded phases for the liquid
chromatographic separation of optical, geometrical, and structural isomers.
4.6 25 22024AST 1 ea
J. Chrom. Sci. 1984, 22 (9), 411-415.
Astec CYCLOBOND® I 2000 Chiral HPLC Guard
Astec CYCLOBOND® Column Screening Kit
For convenience, the four Astec CYCLOBOND CSPs we recommend in the particle size 5 μm
screening protocol described earlier in this section are available in a kit. A full
1.0 2 21010AST 1 ea
description of the screening procedure and techniques to optimize the
4.0 2 21100AST 1 ea
separation are included with each kit.
Kit components: The 4 mm I.D. guard cartridge requires a holder, Cat. No. 21150AST (stand-
• Astec CYCLOBOND I 2000 alone) or 504254 (direct-connect), sold separately. The 1 mm I.D. guard is
• Astec CYCLOBOND I 2000 DMP sold complete and does not require a separate holder.
• Astec CYCLOBOND I 2000 HP-RSP
• Astec CYCLOBOND I 2000 DNP Astec CYCLOBOND® I 2000 AC (Acetyl β-Cyclodextrin)
• Astec CYCLOBOND Handbook
Astec CYCLOBOND I 2000 AC is used primarily for aromatic alcohols or
Also, you can further expand the screening field by incorporating the Astec amines that are chiral on the α or β carbon.
CHIROBIOTIC®, Astec Cellulose DMP, Astec P-CAP, Astec CLC, and protein-
• Bonded phase: Acetylated β-cyclodextrin
based CSPs (all sold separately) into your screening protocol.
• Operating pH range: 3 - 7
Astec CYCLOBOND® HPLC Column Screening Kit • Particle type: High-purity, spherical silica
• Particle diameter: 5 or 10 μm
Ref: 1. Armstrong, D.W., DeMond, W., Cyclodextrin Bonded Phases for the Liquid Chromatographic • Pore size: 100 Å
Separation of Optical, Geometrical, and Structural Isomers J. Chromatogr. Sci. 22, 411 (1984)
• USP Code L45
Description Cat. No. Qty
Astec CYCLOBOND® HPLC Column 20005AST 1 kit For other column dimensions not listed, please inquire.
Screening Kit, particle size 5 μm, L 25 cm ×
I.D. 4.6 mm


Astec CYCLOBOND® Chiral HPLC Columns: Astec CYCLOBOND® I 2000 AC (Acetyl β-Cyclodextrin)
Astec CYCLOBOND® I 2000 AC Chiral HPLC Column Astec CYCLOBOND® I 2000 DMP (Dimethylphenyl
I.D. (mm) L (cm) Cat. No. Qty β-Cyclodextrin)
particle size 5 μm
The reaction of the 3,5-dimethylphenyl isocyanate with the hydroxyl groups
2.1 15 20119AST 1 ea
of β-cyclodextrin results in a pi-basic phase similar in character to the
4.6 5 20121AST 1 ea
naphthylethyl carbamate phases. The selectivity is greater for the Astec
4.6 10 20122AST 1 ea
CYCLOBOND I 2000 DMP when the chiral center of the analyte is part of a
4.6 15 20123AST 1 ea ring structure or is on the α carbon. This phase has been very useful for
4.6 25 20124AST 1 ea derivatized amines, like amphetamine ACQ.
10.0 25 20134AST 1 ea
• Bonded phase: 3,5-Dimethylphenyl carbamate modified β-cyclodextrin
30.0 25 20154AST
particle size 10 μm • Particle type: High-purity, spherical silica
4.6 25 22124AST 1 ea • Particle diameter: 5 or 10 μm
• Pore size: 100 Å
Astec CYCLOBOND® I 2000 AC Chiral HPLC Guard • USP Code L45
I.D. (mm) L (cm) Cat. No. Qty For other column dimensions not listed, please inquire.
particle size 5 μm
1.0 2 21011AST 1 ea Astec CYCLOBOND® I 2000 DMP Chiral HPLC Column
4.0 2 21101AST 1 ea
particle size 5 μm
2.1 15 20719AST 1 ea
sold complete and does not require a separate holder. 2.1 25 20720AST 1 ea
4.6 5 20721AST 1 ea
4.6 10 20722AST 1 ea
Astec CYCLOBOND® I 2000 DM (Dimethyl 4.6 15 20723AST 1 ea
β-Cyclodextrin) 4.6 25 20724AST 1 ea
Astec CYCLOBOND I 2000 DM separates a wide variety of structural and 10.0 25 20734AST 1 ea
geometric isomers and is complementary to Astec CYCLOBOND I 2000. This 21.2 25 20744AST 1 ea
phase operates only in the reversed-phase mode with steric bulk as the 30.0 25 20754AST
main mechanism. particle size 10 μm
• Bonded phase: Dimethylated β-cyclodextrin 4.6 25 22724AST 1 ea

• Particle type: High-purity, spherical silica Astec CYCLOBOND® I 2000 DMP Chiral HPLC Guard
• Particle diameter: 5 or 10 μm I.D. (mm) L (cm) Cat. No. Qty
• Pore size: 100 Å particle size 5 μm
• USP Code L45
1.0 2 21017AST 1 ea
For other column dimensions not listed, please inquire. 4.0 2 21107AST 1 ea
Astec CYCLOBOND® I 2000 DM Chiral HPLC Column The 4 mm I.D. guard cartridge requires a holder, Cat. No. 21150AST (stand-
particle size 5 μm
2.1 15 20919AST 1 ea
4.6 5 20921AST 1 ea
Astec CYCLOBOND® I 2000 DNP (Dinitrophenyl
4.6 10 20922AST 1 ea β-Cyclodextrin)
4.6 15 20923AST 1 ea
This Astec CYCLOBOND derivative has dinitrophenyl functionality bonded
4.6 25 20924AST 1 ea through an ether linkage to the hydroxyl positions of the β-cyclodextrin. In
30.0 25 20954AST this arrangement, a pi-electron sharing system is established with analytes
having pi-electron systems (e.g. aromatic rings, carbonyl) in the stereogenic
Astec CYCLOBOND® I 2000 DM Chiral HPLC Guard environment. Use of the ether linkage to anchor this pi-acidic dinitrophenyl
ring results in a very stable system even under strong reversed-phase
conditions. The pi-acidity of this group is further enhanced with the
particle size 5 μm
introduction of the trifluoromethyl group into the aromatic ring.
1.0 2 21019AST 1 ea
4.0 2 21109AST 1 ea • Bonded phase: Dinitrophenyl modified β-cyclodextrin
The 4 mm I.D. guard cartridge requires a holder, Cat. No. 21150AST (stand- • Particle type: High-purity, spherical silica
alone) or 504254 (direct-connect), sold separately. The 1 mm I.D. guard is • Particle diameter: 5 or 10 μm
sold complete and does not require a separate holder. • Pore size: 100 Å
• USP Code L45


Astec CYCLOBOND® Chiral HPLC Columns: Astec CYCLOBOND® I 2000 DNP (Dinitrophenyl β-Cyclodextrin)
Astec CYCLOBOND® I 2000 DNP Chiral HPLC Column Astec CYCLOBOND® I 2000 RSP (R,S-Hydroxypropyl
I.D. (mm) L (cm) Cat. No. Qty β-Cyclodextrin)
particle size 5 μm
The hydroxyl groups on the surface of the β-cyclodextrin are reacted with
2.1 10 25018AST 1 ea
(R,S)-propylene oxide to yield a general purpose chiral stationary phase. It
2.1 15 25019AST 1 ea
has the added property of separating non-aromatic structures such as t-boc
4.6 5 25021AST 1 ea amino acids, for which it is a standard methodology.
4.6 15 25023AST 1 ea
• Bonded phase: (R,S)-Hydroxypropyl modified β-cyclodextrin
4.6 25 25024AST 1 ea
Astec CYCLOBOND® I 2000 DNP Chiral HPLC Guard
I.D. (mm) L (cm) Cat. No. Qty • Pore size: 100 Å
particle size 5 μm • USP Code L45
1.0 2 25101AST 1 ea For other column dimensions not listed, please inquire.
4.0 2 25100AST 1 ea
Astec CYCLOBOND® I 2000 RSP Chiral HPLC Column
alone) or 504254 (direct-connect), sold separately. The 1 mm I.D. guard is I.D. (mm) L (cm) Cat. No. Qty
sold complete and does not require a separate holder. particle size 5 μm
2.1 10 20318AST 1 ea
2.1 15 20319AST 1 ea
Astec CYCLOBOND® I 2000 HP-RSP
2.1 25 20320AST 1 ea
(R,S-Hydroxypropyl β-Cyclodextrin) 4.6 5 20321AST 1 ea
In the design of this phase chemistry, it was an objective to create a very 4.6 10 20322AST 1 ea
stable and reproducible phase with shorter retention times, while 4.6 15 20323AST 1 ea
maintaining or improving selectivity over Astec CYCLOBOND I 2000 RSP. 4.6 25 20324AST 1 ea
With that goal and more achieved, Astec CYCLOBOND I 2000 HP-RSP 10.0 25 20334AST 1 ea
separates by extended H-bonding capability, and offers broad chiral 21.2 25 20344AST 1 ea
selectivity for chiral screening. It is most beneficial for basic and neutral 30.0 25 20354AST
compounds. particle size 10 μm
• Bonded phase: (R,S)-Hydroxypropyl modified β-cyclodextrin (high 4.6 25 22324AST 1 ea
performance)
• Operating pH range: 3 - 7 Astec CYCLOBOND® I 2000 RSP Chiral HPLC Guard
• Particle diameter: 5 or 10 μm I.D. (mm) L (cm) Cat. No. Qty
• Pore size: 100 Å particle size 5 μm
• USP Code L45 1.0 2 21013AST 1 ea
4.0 2 21103AST 1 ea
Astec CYCLOBOND® I 2000 HP-RSP Chiral HPLC Column
I.D. (mm) L (cm) Cat. No. Qty sold complete and does not require a separate holder.
particle size 5 μm
2.1 10 24018AST 1 ea Astec CYCLOBOND® I 2000 SP (S-Hydroxypropyl
2.1 25 24020AST 1 ea
β-Cyclodextrin)
4.6 5 24021AST 1 ea
4.6 10 24022AST 1 ea On Astec CYCLOBOND I 2000 SP, the hydroxyl groups on the surface of the
4.6 15 24023AST 1 ea β-cyclodextrin have been reacted with (S)-propylene oxide. This has the
4.6 25 24024AST 1 ea effect of extending hydrogen-bonding capabilities to accommodate
analytes with chiral centers that are relatively distant from an aromatic ring
30.0 25 24054AST
structure. The (S)- form shows enhanced selectivity and efficiency for some
separations.
4.6 25 24124AST 1 ea
• Bonded phase: (S)-Hydroxypropyl modified β-cyclodextrin
Astec CYCLOBOND® I 2000 HP-RSP Chiral HPLC Guard • Operating pH range: 3 - 7
I.D. (mm) L (cm) Cat. No. Qty • Particle diameter: 5 or 10 μm
particle size 5 μm • Pore size: 100 Å
1.0 2 24101AST 1 ea • USP Code L45
4.0 2 24100AST 1 ea


Astec CYCLOBOND® Chiral HPLC Columns: Astec CYCLOBOND® I 2000 SP (S-Hydroxypropyl β-Cyclodextrin)
Astec CYCLOBOND® I 2000 SP Chiral HPLC Column Astec CYCLOBOND® II AC (Acetyl γ-Cyclodextrin)
I.D. (mm) L (cm) Cat. No. Qty Astec CYCLOBOND II AC columns are bonded γ-cyclodextrin with
particle size 5 μm acetylation of the 2- and 3-hydroxyl groups. As a result, the mouth of the
2.1 15 20219AST 1 ea cavity has available a hydrogen-acceptor site that can interact with a
4.6 5 20221AST 1 ea hydrogen donor, such as an amine attached to at least two or more fused
4.6 25 20224AST 1 ea rings. An example would be 1- or 2-substituted napthylethylamine.
30.0 25 20254AST Applications include steroids and sterols, depending on where the hydroxyl
particle size 10 μm groups are positioned.
4.6 25 22224AST 1 ea • Bonded phase: Acetylated γ-cyclodextrin
Astec CYCLOBOND® I 2000 SP Chiral HPLC Guard • Particle type: High-purity, spherical silica
I.D. (mm) L (cm) Cat. No. Qty • Pore size: 100 Å
particle size 5 μm
4.0 2 21102AST 1 ea For other column dimensions not listed, please inquire.
The 4 mm I.D. guard cartridge requires a holder, Cat. No. 21150AST (stand- Astec CYCLOBOND® II AC Chiral HPLC Column
alone) or 504254 (direct-connect), sold separately. The 1 mm I.D. guard is I.D. (mm) L (cm) Cat. No. Qty
sold complete and does not require a separate holder. particle size 5 μm
2.1 10 47018AST 1 ea
Astec CYCLOBOND® II (γ-Cyclodextrin) 2.1 15 47019AST 1 ea
Consisting of eight glucopyranose units arranged in a truncated cone shape, 4.6 25 41022AST 1 ea
Astec CYCLOBOND II is an excellent chiral selector for multi-ring structures. It particle size 10 μm
is useful for isomeric compounds based on anthracene, chrysene and 4.6 25 44124AST 1 ea
pyrene type ring structures. Astec CYCLOBOND II offers good selectivity and
stability and is applicable to the polar organic mode of separation. The 4 mm I.D. guard cartridge requires a holder, Cat. No. 21150AST (stand-
Applications include steroids, porphyrins, FMOC amino acids. alone) or 504254 (direct-connect), sold separately. The 1 mm I.D. guard is
• Bonded phase: Underivatized, native γ-cyclodextrin
• Particle type: High-purity, spherical silica Astec Cellulose DMP
• Pore size: 100 Å (Dimethylphenylcarbamate)
For other column dimensions not listed, please inquire. 8P R O D U C T S
Astec CYCLOBOND® II Chiral HPLC Column
Efficient, Rugged and Economical Columns for Chiral HPLC & SFC
I.D. (mm) L (cm) Cat. No. Qty Astec Cellulose DMP comprises spherical, high-purity porous silica coated
particle size 5 μm with DMPC (dimethylphenyl carbamate)-derivatized cellulose packed in
2.1 15 46019AST 1 ea analytical to preparative size HPLC columns. It separates a wide range of
4.6 5 46021AST 1 ea chiral compounds under normal phase, polar organic, SFC, and reversed-
4.6 10 40020AST 1 ea phase conditions, with high efficiency, high loading capacity, and excellent
4.6 15 46023AST 1 ea column lifetime. With performance comparable to other DMPC-derivatized
4.6 25 41020AST 1 ea cellulose CSPs, but at substantially lower price, Astec Cellulose DMP is a
particle size 10 μm must-have for every chiral column HPLC or SFC screening protocol. Astec
4.6 25 44024AST 1 ea
Cellulose DMP is complementary to the other Astec CSPs, including
CHIROBIOTIC<REFERENCE ID="3826" TYPE="trademark"/>, CYCLOBOND, and
the P-CAP product lines. It should be investigated as an alternative to higher
Astec CYCLOBOND® II Chiral HPLC Guard
priced cellulose-DMPC columns for existing methods. The cost savings are
I.D. (mm) L (cm) Cat. No. Qty especially dramatic when comparing preparative column dimensions.
particle size 5 μm • Phase: DMPC (dimethylphenyl carbamate)-derivatized cellulose (coated)
• Particle diameter: 5 μm
The 4 mm I.D. guard cartridge requires a holder, Cat. No. 21150AST (stand- • Normal phase, polar organic, and SFC modes
alone) or 504254 (direct-connect), sold separately. The 1 mm I.D. guard is • Scalable from analytical to preparative
sold complete and does not require a separate holder. • Suitable for USP Code L40
Related Information
No. Title
T410110 Astec Cellulose DMP Brochure


Astec Cellulose DMP (Dimethylphenylcarbamate)
Astec Cellulose DMP Chiral HPLC Column (1) Gasparrini, F.; Misiti, D.; Rompietti, R; Villani, C. "New hybrid polymeric
I.D. (mm) L (cm) Cat. No. Qty liquid chromatography chiral stationary phase prepared by surface-initiated
particle size 5 μm polymerization" J. Chromatogr. A 2005, 1064 (1), 25-38.
2.1 10 51112AST 1 ea
(2) Zhong, Q.; Han, X.; He, L.; Beesley, T. E.; Trahanovsky, W. S.; Armstrong, D.
W. "Chromatographic evaluation of poly(trans-1,2-cyclohexanediyl-bisacry-
2.1 15 51100AST 1 ea
lamide) as a chiral stationary phase for HPLC" J. Chromatogr. A 2005, 1066 (1-
2.1 25 51101AST 1 ea
2), 55-70.
4.6 10 51097AST 1 ea
4.6 15 51098AST 1 ea
4.6 25 51099AST 1 ea Related Information
10 25 51102AST 1 ea Request free literature by phone, fax, or visit sigma-aldrich.com/literature.
21.2 25 51103AST 1 ea
No. Title
T410060 Astec P-CAP™ and P-CAP™-DP Brochure
Astec Cellulose DMP Chiral HPLC Guard
particle size 5 μm Astec (R,R) and (S,S) P-CAP™
2.1 2 51105AST 1 kit
Astec P-CAP is made from a polymerized diacryloyl-trans-1,2-diphenylethy-
2.1 2 51104AST 2 ea
lenediamine bonded to the silica surface. It utilizes hydrogen bonding and
4 2 51107AST 1 kit
steric effects as enantiomer separation mechanisms. Astec P-CAP can be
4 2 51106AST 2 ea
used for SFC and normal phase separations of racemic mixtures. It has high
10 1 51108AST 1 ea stability, high sample loading capacity (suitable for preparative scale-up),
21.2 1 51109AST 1 ea and no memory effect. The elution order of compounds can be reversed in
the (R,R) versus (S,S) configuration.
Guard cartridges require holders that are sold separately. The 2.1 and 4 mm
I.D. cartridges use 21150AST or 59660-U (both stand-alone) or 504254 or • Bonded phase: Poly(trans-1,2-cyclohexanediyl-bis-acrylamide)
55205 (both integral). The 10 mm I.D. cartridges use 567499-U. The 21.2 mm • Operating pH range: N/A (operated in normal phase and polar organic
I.D. cartridges use 581392-U. modes)
• Particle diameter: 3.5, 5 or 10 μm
Astec P-CAP™ and P-CAP™-DP Chiral HPLC • Pore size: 200 Å
Columns For other column dimensions not listed, please inquire.
Useful for chiral HPLC and SFC separations, Astec P-CAP and P-CAP-DP are Astec (R,R) P-CAP™ Chiral HPLC Column
based on a unique polycyclic amine polymer that has been covalently
bonded to high-purity silica particles. They offer high stability, extremely I.D. (mm) L (cm) Cat. No. Qty
high sample loadability, easy scale-up and no memory effect. Concep- particle size 3.5 μm
tualized by Prof. Francesco Gasparrini (1) with further phase development by 4.6 15 30023AST 1 ea
Prof. Daniel Armstrong (2), these CSPs are used primarily for normal phase, particle size 5 μm
polar organic and SFC chiral separations. The bonding procedure offers 2.1 15 31019AST 1 ea
maximum protection of the silica and excellent availability of the short-chain 4.6 5 31021AST 1 ea
polymer ligand to ensure high capacity. The resulting thin, ordered layer of 4.6 10 31022AST 1 ea
polymer does not alter the porous structure of the silica. The repeating chiral 4.6 15 31023AST 1 ea
moiety offers both structural conformation and hydrogen bonding 4.6 25 31024AST 1 ea
interactions as the driving mechanisms. particle size 10 μm
Preparative separations on Astec P-CAP and P-CAP-DP can be run in a 4.6 25 31124AST 1 ea
variety of solvents, without any large impact on selectivity, to meet analyte
solubility requirements. As a result of the juxtaposition of the binding sites, The 4 mm I.D. guard cartridge requires a holder, Cat. No. 21150AST (stand-
molecules with two or more functional groups demonstrate the best alone) or 504254 (direct-connect), sold separately. The 1 mm I.D. guard is
selectivity. Separations have been run in pure acetone, heptane/ethanol, sold complete and does not require a separate holder.
dichloromethane/methanol and ethylacetate. Selectivity can be obtained in
a variety of solvent choices with different efficiencies. Salt and/or acetic acid Astec (S,S) P-CAP™ Chiral HPLC Column
can be added to improve efficiency or enhance detection in mass
spectrometry. Astec P-CAP and P-CAP-DP are available in two enantiomeric
particle size 3.5 μm
forms (R,R) and (S,S). This permits reversing the elution order, which can be
very useful in preparative applications. 4.6 5 32021AST 1 ea
4.6 15 32023AST 1 ea
Astec P-CAP and P-CAP-DP features and application areas: particle size 5 μm
2.1 10 33018AST 1 ea
• Polymeric ligand CSP for normal phase and polar organic operation
• Ideal for SFC and Sub-SFC 2.1 15 33019AST 1 ea
• No solvent limitations 4.6 5 33021AST 1 ea
• High capacity for preparative applications 4.6 10 33022AST 1 ea
• Stable, covalent chemistry 4.6 15 33023AST 1 ea
• Reversible elution order through R,R and S,S configurations 4.6 25 33024AST 1 ea
• Available in standard (Astec P-CAP) and diphenyl (Astec P-CAP-DP) particle size 10 μm
chemistries 4.6 25 33124AST 1 ea


Astec P-CAP™ and P-CAP™-DP Chiral HPLC Columns: Astec (R,R) and (S,S) P-CAP™
Astec (S,S) P-CAP™ Chiral HPLC Guard Column Astec CLC-L and CLC-D (Copper Ligand
I.D. (mm) L (cm) Cat. No. Qty Exchange)
particle size 5 μm
4.0 2 33100AST 1 ea Astec CLC columns use the copper ligand concept described by Davankov
to effect enantiomer separation (1,2). The method uses a small, chiral
The 4 mm I.D. guard cartridge requires a holder, Cat. No. 21150AST (stand- bidentate ligand attached to the silica surface and a copper sulphate-
alone) or 504254 (direct-connect), sold separately. The 1 mm I.D. guard is containing mobile phase. The copper ions coordinate with the chiral
sold complete and does not require a separate holder. selector on the stationary phase and carboxylic acid functional groups on
the analytes to form transient diastereomeric complexes in solution. The
technique also has the advantage of giving small acids with no UV
Astec (R,R) and (S,S) P-CAP™-DP chromophore a strong 254 nm signal.
The DP introduces phenyl rings to add pi-pi interactions, giving it one Astec CLC columns are ideal for analysis of alpha-hydroxy acids, like lactic,
additional type of interaction compared to Astec P-CAP. Astec P-CAP-DP malic, tartaric and mandelic acids, amino acids, other amines and bi-
uses similar protocols as Astec P-CAP and can be optimized for either functional racemates, like amino alcohols. Two versions of the column
normal or polar organic mobile phases. It is less polar than Astec P-CAP, and provide elution order reversal. On the CLC-D column, the L enantiomer
ideal for sub- and supercritical fluid applications. The elution order of generally elutes before D, with the exception of tartaric acid. The reverse is
compounds can be reversed in the (R,R) versus (S,S) configuration. true on the CLC-L column where D elutes before L. Proline and aspartic acid
• Bonded phase: Poly(diphenylethylenediamine-bis-acryloyl) or Poly-DPEDA are particularly suited for low-level detection on the CLC column since the
• Operating pH range: N/A (operated in normal phase and polar organic copper complex is detected at 254 nm UV. Both can be resolved on the
modes) CLC-D or CLC-L in 5 mM CuSO4 with the usual reversal of elution order from
• Particle type: High-purity, spherical silica the CLC-D to CLC-L. In theory, any analyte that can complete the
• Particle diameter: 3.5 or 5 μm coordination with the copper ion can be resolved.
• Pore size: 200 Å Astec CLC features and application areas:
For other column dimensions not listed, please inquire. • Separates α-hydroxy carboxylic acids, amino acids and other α-bifunc-
tional compounds
Astec (R,R) P-CAP™-DP Chiral HPLC Column • High selectivity with simple mobile phases
• Copper complex gives strong UV 254 nm signal
I.D. (mm) L (cm) Cat. No. Qty • Simple reversal of elution order, AstecCLC-L vs. AstecCLC-D
particle size 3.5 μm • Excellent reproducibility
4.6 15 34023AST 1 ea
particle size 5 μm
Properties of Astec CLC-L and Astec CLC-D:
4.6 15 35023AST 1 ea • Bonded phase: Chiral bidentate ligand (L and D forms)
4.6 25 35024AST 1 ea • Requires 5 mM CuSO4 mobile phase
21.2 25 35044AST 1 ea • Operating pH range: 3 - 6 (adjust pH of the 5 mM CuSO4 mobile phase
with acetic acid)
Astec (R,R) P-CAP™-DP Chiral HPLC Guard Column • Particle type: High-purity spherical silica
I.D. (mm) L (cm) Cat. No. Qty • Pore size: 100Å
particle size 5 μm • USP Code L32
4.0 2 35100AST 1 ea
(1) Davankov, V. A.; Rogozhin, S. V. Ligand chromatography as a novel
method for the investigation of mixed complexes: Stereoselective effects in
α-amino acid copper(II) complexes. J. Chrom. A. 1971, 60, 280-283.
alone) or 504254 (direct-connect), sold separately.
(2) Davankov, V. A. Enantioselective ligand exchange in modern separation
techniques. J. Chrom. A. 2003, 1000, 891-915.
Astec (S,S) P-CAP™-DP Chiral HPLC Column
particle size 3.5 μm Related Information
4.6 15 36023AST 1 ea Request free literature by phone, fax, or visit sigma-aldrich.com/literature.
particle size 5 μm No. Title
4.6 15 37023AST 1 ea T410062 Astec CLC (Copper Ligand Exchange) Flyer
4.6 25 37024AST 1 ea
The 4 mm I.D. guard cartridge requires a holder, Cat. No. 21150AST (stand- Astec CLC-D Chiral HPLC Column
alone) or 504254 (direct-connect), sold separately.
particle size 5 μm
4.6 15 53023AST 1 ea
Astec CLC-L Chiral HPLC Column

particle size 5 μm
4.6 15 53123AST 1 ea
We recommend using a Supelco ColumnSaver precolumn filter (Cat. No.

55214-U or 55215-U) to protect CLC columns.


Astec CLC-L and CLC-D (Copper Ligand Exchange)
HPLC Analysis of Lactic Acid Enantiomers on Astec® CLC-L and CHIRALPAK® AGP (α1-Acid Glycoprotein)
CLC-D
CHIRALPAK® AGP has the broadest range of selectivity of all protein phases
 application for HPLC currently available. It comprises α1-acid glycoprotein (AGP) as the chiral
column ....... Astec CLC-L and CLC-D, 15 cm x 4.6 mm I.D., 5 μm particles (53023AST, 53123AST) selector immobilized onto spherical 5 μm silica particles. When bonded,
mobile phase ................................................................................................................................................................... 5 mM CuSO4
flow rate ...................................................................................................................................................................................... 1 mL/min
AGP is very stable and tolerates pure organic solvents (up to 20%), elevated
column temp. .................................................................................................................................................................................... 25 °C temperatures (up to 40 °C), and pH values from 4 to 7. Operated in reversed-
detector ............................................................................................................................................................................. UV at 254 nm phase mode, CHIRALPAK® AGP separates enantiomers of an extremely
injection ................................................................................................................................................................................................. 10 μL broad range of drug substances, such as acids, amines and neutral
sample .......................................................................................................................................................................................... lactic acid
compounds. The mobile phases are mixtures of phosphate or acetate
Application No. .................................................................................................................................................................... G004399
buffers and organic solvents such as 2-propanol or acetonitrile. The
enantioselectivity and retention can easily be regulated by mobile phase pH
CLC-L CLC-D and ionic strength, and the nature and concentration of the organic
(D)-Lactic acid, (L)-Lactic acid, modifier. The most important tool in method development is the mobile
9.24 min 9.27 min phase pH, which affects the ionization of both solutes and the protein
stationary phase. AGP has a low isoelectric point (pI) of 2.7. This means at pH
(L)-Lactic acid, (D)-Lactic acid, 2.7 the column has a net zero charge. From pH 2.7 to 7, the net negative
12.58 min 12.18 min
charge on the AGP molecule increases, providing increased retention of
positively-charged analytes, like amines. These compounds are also retained
O by hydrophobic and hydrogen bonding interactions.
H3C • Bonded phase: α1-Acid glycoprotein (CHIRALPAK® AGP)
OH • Particle type: High-purity spherical silica
OH • Particle diameter: 5 μm
• Maximum organic percentage in mobile phase: 20%
• Maximum pressure: 2000 psi
• Maximum operating temperature: 40 °C
• Washing procedure: 10-15% isopropanol in water (do not exceed max.
pressure)
• USP Code L41
CHIRALPAK® AGP HPLC Column

Protein-based Chiral HPLC Columns I.D. (mm) L (cm) Cat. No. Qty
Hermansson described the use of natural proteins immobilized onto a silica particle size 5 μm
support for chiral separations in 1983 (1). Proteins contain a large number of 2.0 5 58129AST 1 ea
chiral centers of one configuration, and many other sites that contribute to 2.0 10 58130AST 1 ea
the general retention process. We offer three CSPs with proteins as the chiral 2.0 15 58131AST 1 ea
selectors, CHIRALPAK® AGP (α1-acid glycoprotein), CHIRALPAK® CBH 3.0 5 58169AST 1 ea
(cellobiohydrolase) and CHIRALPAK® HSA (human serum albumin). All are 3.0 10 58170AST 1 ea
manufactured by DAICEL Corporation. They are typically used in reversed- 3.0 15 58171AST 1 ea
phase mode, and perform a wide variety of chiral separations. CHIRALPAK® 4.0 5 58149AST 1 ea
HSA is also used for drug-binding studies. Solutes are retained by three 4.0 10 58150AST 1 ea
types of interactions: ionic (for charged solutes), hydrophobic, and hydrogen
4.0 15 58151AST 1 ea
bonding. The relative contribution of the different forces to solute retention
10.0 10 58155AST 1 ea
depends on the nature of the analyte.
10.0 15 58157AST 1 ea
CHIRALPAK® AGP: Extremely broad applicability. First choice when
developing methods on protein-CSPs. CHIRALPAK® AGP HPLC Guard Column
CHIRALPAK® HSA: Analytes are typically very hydrophilic acids.
CHIRALPAK® CBH: Analytes are typically very hydrophilic amines and amino I.D. (mm) L (cm) Cat. No. Qty
alcohols. particle size 5 μm
Protein-based CSP features and application areas: 2.0 1 58178AST 2 ea
3.0 1 58158AST 2 ea
• Direct reversed-phase resolution of chiral molecules
4.0 1 58188AST 2 ea
• Stable in a variety of organic modifiers
• Available in analytical and semi-preparative sizes
The 2, 3, and 4 mm I.D. guard cartridges require a guard column holder (Cat.
• CHIRALPAK HSA is also used for drug-binding studies
No. 58159AST), sold separately.
(Note: These columns were previously named CHIRAL-AGP, CHIRAL-HSA,
and CHIRAL-CBH prior to the acquisition of ChromTech by DAICEL Corp.)
(1) Hermansson, J. Direct liquid chromatographic resolution of racemic
drugs using α1-acid glycoprotein as the chiral stationary phase. J.
Chromatogr. A, 1983, 269, 71-80.


Protein-based Chiral HPLC Columns: CHIRALPAK® AGP (α1-Acid Glycoprotein)
Guard Column Holder for CHIRALPAK® AGP, HSA, and CBH CHIRALPAK® CBH HPLC Guard Column
 for use with CHIRALPAK AGP, CBH, and HSA 1 cm x 2.0, 3.0, and 4.0 mm I.D. (mm) L (cm) Cat. No. Qty
guard cartridges particle size 5 μm
stainless steel 2.0 1 58578AST 2 ea
3.0 1 58558AST 2 ea
4.0 1 58588AST 2 ea
The 2, 3, and 4 mm I.D. guard cartridges require a guard column holder (Cat.
No. 58159AST), sold separately.
Holder for CHIRALPAK AGP, HSA, and CBH 1 cm length guard cartridges (58159AST)
CHIRALPAK® HSA (Human Serum Albumin)
CHIRALPAK® HSA, which uses human serum albumin as the chiral selector, is
highly selective for acidic racemates, preferably weak and strong acids,
zwitterionic and non-protolytic (neutral) compounds. Phosphate buffers
(normally 0.01-0.1M, pH 5-7) with addition of organic modifiers are used as
mobile phases. Enantioselectivity and retention can be regulated by
changing the mobile phase composition. However, the primary use of
Right, holder (58159AST) and left, representative CHIRALPAK® AGP, CHIRALPAK® HSA is for fast drug/protein binding studies (1). To calculate the
HSA, or CBH 1 cm length guard cartridges.
% protein binding, measure the retention time of an unretained compound
58159AST 1 ea (t0) and the compound of interest (tr) on the CHIRALPAK® HSA column. Then
use the capacity factor equation:
k = (tr - t0)/tr
CHIRALPAK® CBH (Cellobiohydrolase) to calculate the % protein binding (P):
P = 100k/(k+1)
Used primarily for the separation enantiomers of basic compounds,
Different types of mobile phases can be used. A mobile phase consisting of
CHIRALPAK® CBH has cellobiohydrolase as the chiral selector immobilized
6% 2-propanol in 20 mM potassium phosphate buffer, pH 7.0 gives data in
on spherical 5 μm silica particles. Used in reversed-phase mode, retention
good agreement with literature data. The mobile phase conditions should
and enantioselectivity is regulated by changes of pH, buffer concentration
be chosen to suit the drugs to be tested, i.e., for high protein binding drugs
and organic modifier. The mobile phases are mixtures of phosphate or
a mobile phase with higher eluting strength might be needed in order to
acetate buffers and organic solvents such as 2-propanol or acetonitrile. The
reduce retention times.
column is preferably used for the separation of enantiomers of basic drugs,
particularly compounds containing one or more nitrogen atoms along with • Bonded phase: Human serum albumin (HSA)
one or more hydrogen-bonding groups (alcohol, phenol, carbonyl, amide, • Particle type: High-purity spherical silica
ether, ester, etc.). • Particle diameter: 5 μm
• Bonded phase: Cellobiohydrolase (CBH)
• Particle type: High-purity spherical silica
• Maximum operating temperature: 40 °C
• Washing procedure: 10-15% isopropanol in water (do not exceed max.
pressure)
• Maximum operating temperature: 40 °C (1) Goodman, A.; Gilman, A.G. The Pharmacological Basis of Therapeutics, 9th
• Washing procedure: 10-15% isopropanol in water (do not exceed max. Edition, McGraw-Hill: New York, 1996; pp 1712-1792.
pressure)
CHIRALPAK® HSA HPLC Column
CHIRALPAK® CBH HPLC Column I.D. (mm) L (cm) Cat. No. Qty
I.D. (mm) L (cm) Cat. No. Qty particle size 5 μm
particle size 5 μm 2.0 5 58429AST 1 ea
2.0 5 58529AST 1 ea 2.0 10 58430AST 1 ea
2.0 10 58530AST 1 ea 2.0 15 58431AST 1 ea
2.0 15 58531AST 1 ea 3.0 5 58469AST 1 ea
3.0 5 58569AST 1 ea 3.0 10 58470AST 1 ea
3.0 10 58570AST 1 ea 3.0 15 58471AST 1 ea
3.0 15 58571AST 1 ea 4.0 5 58449AST 1 ea
4.0 5 58549AST 1 ea 4.0 10 58450AST 1 ea
4.0 10 58550AST 1 ea 4.0 15 58451AST 1 ea
4.0 15 58551AST 1 ea 10.0 10 58455AST 1 ea
10.0 10 58555AST 1 ea 10.0 15 58457AST 1 ea
10.0 15 58557AST 1 ea


Protein-based Chiral HPLC Columns: CHIRALPAK® HSA (Human Serum Albumin)
CHIRALPAK® HSA HPLC Guard Column Kromasil® AmyCoat® Chiral HPLC Column
I.D. (mm) L (cm) Cat. No. Qty I.D. (mm) L (mm) Cat. No. Qty
particle size 5 μm particle size 3 μm
2.0 1 58478AST 2 ea 2.1 50 K08971229 1 ea
3.0 1 58458AST 2 ea 2.1 150 K08971225 1 ea
4.0 1 58488AST 2 ea 4.6 50 K08670344 1 ea
4.6 150 K08670346 1 ea
The 2, 3, and 4 mm I.D. guard cartridges require a guard column holder (Cat. particle size 5 μm
No. 58159AST), sold separately. 2.1 50 K08971230 1 ea
2.1 150 K08971226 1 ea
Kromasil® Chiral HPLC Columns 4.6 50 K08670608 1 ea

4.6 150 K08670347 1 ea
8P R O D U C T S 4.6 250 K08670348 1 ea
10 250 K08670605 1 ea
We are pleased to be able to offer Kromasil products, including their high- 21.2 250 K08670606 1 ea
quality chiral HPLC line, to our customers in the USA (including Puerto Rico) 30 250 K08670607 1 ea
and Canada. Kromasil chiral stationary phases have an excellent reputation particle size 10 μm
in analytical to process scale HPLC and SFC. 4.6 150 K08670603 1 ea
Kromasil AmyCoat® and CelluCoat® 4.6 250 K08670604 1 ea
10 250 K08670600 1 ea
The functionalized amylose and cellulose coated chiral selectors are coated
21.2 250 K08670601 1 ea
onto a wide pore silica (>1000 Å) matrix; this silica has a low surface area,
30 250 K08670602 1 ea
which reduces the number of achiral interaction sites and thus increases the
chiral selectivity. High resolution, excellent selectivity, high-pressure stability,
and stable performance when switching between compatible mobile Kromasil® AmyCoat® Chiral Reversed Phase HPLC Column
phases are some important benefits. AmyCoat and CelluCoat columns are I.D. (mm) L (mm) Cat. No. Qty
also available in columns that are compatible with reversed-phase particle size 3 μm
operation. 4.6 50 K08670500 1 ea
• AmyCoat: The chiral selector is tris-(3,5-dimethylphenyl)carbamoyl 4.6 150 K08670501 1 ea
amylose (USP Code L51)
• CelluCoat: The chiral selector is tris-(3,5-dimethylphenyl)carbamoyl Kromasil® AmyCoat® Chiral HPLC Guard Cartridge
cellulose (USP Code L40)
I.D. (mm) L (mm) Cat. No. Qty
Kromasil DMB and TBB particle size 5 μm
Kromasil DMB and TBB bonded chiral phases separate a broad range of 4.6 10 K08971102 1 ea
racemates. These 2 phases have been developed to complement each other 10 10 K08971103 1 ea
in selectivity. The chiral monomers are polymerized with a hydrosilane to 21.2 10 K08971104 1 ea
yield a network polymer, which incorporates the bifunctional C2-symmetric
chiral selector and is covalently bonded onto 100 Å silica. Kromasil guard cartridges require a holder and coupler that are sold
• Chiral DMB: The chiral monomer is O,O′-bis (3,5-dimethylbenzoyl)- separately. For 2.1 to 4.6 mm I.D. cartridges: Use holder K08970954 and
N,N′-diallyl-L-tartar diamide coupler K08970955. For 10 to 21.2 mm I.D. cartridges: Use holder
• Chiral TBB: The chiral monomer is O,O′-bis (4-tert-butylbenzoyl)- K08970956 and coupler K08970957.
N,N′-diallyl-L-tartar diamide
Kromasil® CelluCoat® Chiral HPLC Column
Kromasil Guard Columns
The Kromasil guards are sold in packs of 3 or 5 cartridges. They require a
particle size 3 μm
holder and coupler that are sold separately.
2.1 50 K08971227 1 ea
• For 2.1 to 4.6 mm I.D. cartridges: Use holder K08970954 and coupler 2.1 150 K08971223 1 ea
K08970955. particle size 5 μm
• For 10 to 21.2 mm I.D. cartridges: Use holder K08970956 and coupler
2.1 50 K08971228 1 ea
K08970957.
2.1 150 K08971224 1 ea
A convenient Guard Cartridge Starter Kit for 4.6 mm I.D. Kromasil CelluCoat particle size 3 μm
columns is available. It contains 5 guard cartridges, a guard cartridge holder, 4.6 50 K08670372 1 ea
and a coupler. The part number is K08971109. 4.6 150 K08670370 1 ea
Related Information
No. Title
T409214 Kromasil® Chiral Applications Guide


Kromasil® Chiral HPLC Columns

particle size 5 μm Cyclofructans: LARIHC™ and FRULIC™ Chiral
4.6 50 K08670617 1 ea HPLC and HILIC Columns
4.6 150 K08670373 1 ea
4.6 250 K08670374 1 ea 8P R O D U C T S
10 250 K08670614 1 ea
21.2 250 K08670615 1 ea Cyclofructans are cyclic oligosaccharides and the newest class of chiral
30 250 K08670616 1 ea stationary phases for HPLC, SFC, and HILIC. Invented by Prof. Daniel W.
particle size 10 μm Armstrong (1,2) and introduced by AZYP, the LARIHC and FRULIC
4.6 150 K08670612 1 ea derivatized cyclofructan-based HPLC columns are now available world-wide
4.6 250 K08670613 1 ea through Supelco/Sigma-Aldrich.
10 250 K08670609 1 ea CF Phase Mode Description
21.2 250 K08670610 1 ea LARIHC CF6-P Chiral HPLC Alkyl derivatized cyclofructan 6. Particularly useful for
30 250 K08670611 1 ea chiral primary amines.
LARIHC CF6-M Chiral HPLC Methyl-functionalized cyclofructan 6. Complementary
Kromasil® CelluCoat® Chiral Reversed Phase HPLC Column to LARIHC CF6-P for chiral primary amines.
LARIHC CF6-RN Chiral HPLC R-Naphthylethyl-functionalized cyclofructan 6.
I.D. (mm) L (mm) Cat. No. Qty Suitable for enantiomers that are not primary amines.
particle size 3 μm LARIHC CF7-DMP Chiral HPLC 3,5-Dimethylphenyl functionalized cyclofructan 7.
4.6 50 K08670502 1 ea Complementary enantioselectivity to LARIHC CF-6-RN.
4.6 150 K08670503 1 ea FRULIC-N HILIC Neutral poly-hydroxy based stationary phase
FRULIC-C HILIC Hydrophilic charged cyclofructan 6
Kromasil® CelluCoat® Chiral HPLC Guard Cartridge
All phases are available in standard HPLC column dimensions. For more
information on AZYP′s LARIHC and FRULIC columns, please visit our website
particle size 5 μm or contact your local Sigma-Aldrich office.
4.6 10 K08971106 1 ea
10 10 K08971107 1 ea
References:
(1) Ping Sun, Chunlei Wang, Zachary S. Breitbach, Ying Zhang, and Daniel W.
21.2 10 K08971108 1 ea
Armstrong. "Development of New HPLC Chiral Stationary Phases Based on
Native and Derivatized Cyclofructans" Anal. Chem. 2009, 81, 10215-10226.
Kromasil guard cartridges require a holder and coupler that are sold
(2) Ping Sun and Daniel W. Armstrong. "Effective enantiomeric separations of
separately. For 2.1 to 4.6 mm I.D. cartridges: Use holder K08970954 and
racemic primary amines by the isopropyl carbamate-cyclofructan6 chiral
coupler K08970955. For 10 to 21.2 mm I.D. cartridges: Use holder
stationary phase" J. Chromatogr. A, 2010, 1217, 4904-4918.
K08970956 and coupler K08970957.
Kromasil guard cartridges require a holder and coupler that are sold
separately. For 2.1 to 4.6 mm I.D. cartridges: Use holder K08970954 and Protection for Chiral HPLC Columns
coupler K08970955. For 10 to 21.2 mm I.D. cartridges: Use holder
K08970956 and coupler K08970957. Within our chiral HPLC line we offer three distinct guard column formats:
• 2 cm x 1.0 mm I.D. stand-alone guard columns for all Astec CSPs
Kromasil® TBB Chiral HPLC Column (CHIROBIOTIC®;, CYCLOBOND, Cellulose, P-CAP)
• 2 cm x 4.0 mm I.D. guard cartridges for all Astec CSPs (holder required)
• 1 cm x 2, 3, and 4 mm I.D. guard cartridges for the protein-based CSPs
particle size 5 μm
(CHIRAL-AGP, -HSA, -CBH) (holder required)
4.6 250 K08670376 1 ea
Examples are shown below. We also offer the Supelco ColumnSaver direct-
Kromasil guard cartridges require a holder and coupler that are sold connect in-line filter (55214-U or 55215-U) to remove particulate matter. The
separately. For 2.1 to 4.6 mm I.D. cartridges: Use holder K08970954 and ColumnSaver can be used to protect any of our HPLC columns. This section
coupler K08970955. For 10 to 21.2 mm I.D. cartridges: Use holder describes the hardware needed for the various guard designs. Packed guard
K08970956 and coupler K08970957. cartridges and columns can be found with the respective CSP they are
intended to protect.
Astec 2 cm x 1 mm I.D. HPLC guard column. Does not require a holder.
Representative 2 cm length Supelguard or Astec HPLC guard cartridge.

Requires a stand-alone (21150AST) or direct-connect (504254) holder.
Representative CHIRALPAK® AGP, HSA, or CBH HPLC guard cartridge,

1 cm length by 2, 3 or 4 mm I.D. Requires holder 58159AST.


Protection for Chiral HPLC Columns: Column Protection for Astec Chiral HPLC Columns (CHIROBIOTIC®, CYCLOBOND®, Cellulose DMP, P-CAP™, CLC)
Column Protection for Astec Chiral HPLC Columns Column Protection for Protein-based HPLC Columns
(CHIROBIOTIC®, CYCLOBOND®, Cellulose DMP, Guards for the CHIRALPAK® AGP, HSA, and CBH columns are supplied in 1
P-CAP™, CLC) cm length by 2.0, 3.0 or 4.0 mm I.D. cartridge format in packs of 2. They
require a holder (58159AST) that is sold separately. The holder accom-
Guards are available for Astec CHIROBIOTIC®;, CYCLOBOND, Cellulose, modates standard 1/16" O.D. tubing. You can couple the holder to the
and P-CAP columns in the following dimensions: analytical column using a short piece of 1/16" tubing, or use the convenient
• 2 cm x 4.0 mm I.D. packed guard cartridges that use both stand-alone column couplers. A list of suggested hardware appears in the table below.
(21150AST) or direct-connect (504254) holders. The choice depends on Our complete hardware offering appears in the HPLC Accessories section of
user preferences. These holders accommodate standard 1/16" O.D. Valco- this catalog.
type nuts and ferrules, and have a freely-rotating inlet/outlet port that
allows for complete rotation of tubing on one side of the holder. The
direct-connect style holder attaches directly to Supelco or Astec 3.0, 4.0
and 4.6 mm I.D. columns.
• 2 cm x 1.0 mm I.D. packed guard columns for protecting 2.1 mm and
lesser I.D. columns. The 1.0 mm I.D. columns do not require a holder.
Both of these guard designs use 1/16" O.D. tubing, nuts and ferrules (not
included). You can couple the stand-alone holders to the analytical column Right, holder (58159AST) and left, representative CHIRALPAK® AGP,
HSA, or CBH 1 cm length guard cartridges.
using a short piece of 1/16" tubing, or use the convenient column couplers.
A list of suggested hardware appears in the table below. Our complete Cat. No. Qty
hardware offering appears in the HPLC Accessories section of this catalog. Coupler for Legacy Guard Column Holder
Other guard dimensions, including preparative guards, are available. Please PEEK, I.D. 0.010 in. × O.D. 1=16 in. × 54986 1 ea
inquire. Overall L 1 in.
Guard Column Holder for CHIRALPAK® AGP, HSA, and CBH
for use with CHIRALPAK AGP, CBH, 58159AST 1 ea
and HSA 1 cm x 2.0, 3.0, and 4.0 mm
guard cartridges
HPLC Column Coupler
PEEK, I.D. 0.007 in. × O.D. 1=16 in. × 58162AST 1 ea
Overall L 1 in.
Stainless Steel 1/16 in. Capillary Tubing
L 5 cm × O.D. 1=16 in. × I.D. 0.007 in. 56713 1 ea
Stand-alone HPLC guard column holder (21150AST) and representative
2 cm length Supelguard or Astec guard cartridge. Stainless Steel HPLC Fittings
ferrule, configured for 1=16 in. tubing 22988 10 ea
nut, for for 1=16 in. tubing 22990-U 10 ea
Pre-column Filters
Our applications chemists have found the Supelco ColumnSaver pre-
Direct-connect style holder (504254) for 2 cm length Supelguard and Astec guard cartridges.
Connects to 3, 4 and 4.6 mm I.D. Supelco or Astec HPLC columns.
column filter to be very good at protecting the column from particulate
matter in the sample and mobile phase. This simple in-line filter comes in
Cat. No. Qty two frit porosities, 0.5 and 2 micron. More information on this product can
Coupler for Legacy Guard Column Holder be found in the HPLC Accessories section.
PEEK, I.D. 0.010 in. × O.D. 1=16 in. × 54986 1 ea
Overall L 1 in. Supelco® ColumnSaver Precolumn Filter
Supelguard™ Guard Cartridge Holder
Stand-Alone (Swivel-type), for use 21150AST 1 ea
with Supelguard cartridges (2 cm L.
x 2 to 4.6 mm I.D.)
Direct-Connect (Swivel-type), for use 504254 1 ea
with Supelguard cartridges (2 cm L.
x 3 to 4.6 mm I.D.)
Stand-Alone, for use with Supel- 567499-U 1 ea
guard cartridges (1 cm L. x 10.0 mm
I.D.)
Stand-Alone, for use with Supel- 581392-U 1 ea Description Cat. No. Qty
guard cartridges (1 cm L. x 21.2 mm
I.D.) 0.5 μm 55214-U 10 ea
Stainless Steel HPLC Fittings 2.0 μm 55215-U 10 ea
ferrule, configured for 1=16 in. tubing 22988 10 ea
nut, for for 1=16 in. tubing 22990-U 10 ea
Stainless Steel 1/16 in. Capillary Tubing
L 5 cm × O.D. 1=16 in. × I.D. 0.007 in. 56713 1 ea
HPLC Column Coupler
PEEK, I.D. 0.007 in. × O.D. 1=16 in. × 58162AST 1 ea
Overall L 1 in.


Chiral HPLC Column Test Mixes
Chiral HPLC Column Test Mixes Chiral GC Columns

Use these test mixes to evaluate the performance of your chiral HPLC The acquisition of Astec by Sigma-Aldrich in 2006 merged two well-
column and make sure it is operating effectively. Consult the QA report established lines of CSPs for enantiomer separations by capillary GC. Both
supplied with the column, or call or email our Technical Services for mobile Supelco DEX™ and Astec CHIRALDEX® are based on cyclodextrins and
phase and expected performance criteria. exhibit complementary selectivity. All columns are manufactured to deliver
• 5-Methyl-5-phenylhydantoin is used to evaluate the performance of Astec high resolution and analyte response, low bleed, and long column life.
CHIROBIOTIC® columns. The mobile phase is 100% methanol and Our chiral GC columns currently comprise:
detection is by UV at 254 nm. The test mix is supplied as a racemic mixture
of two enantiomers. • Astec CHIRALDEX® - Developed by Astec, the CHIRALDEX line of chiral
• Trans-stilbene oxide (TSO) is used to evaluate the performance of Astec capillary GC columns use specialized phase chemistries that include
Cellulose DMP and other polysaccharide-based chiral HPLC columns. The unique derivatives of cyclodextrins with a broad range of selectivities. The
recommended mobile phase is 10:90 IPA:hexane and detection is by UV at "TA" (trifluoroacetyl) derivatives possess the most popular and unique
220 nm. The test mix is supplied as a racemic mixture of the two TSO chemistry.
enantiomers and 1,3,5-tri-tert-butylbenzene is a void volume marker. • Supelco DEX™ - Developed by Supelco, DEX capillary GC columns
comprise derivatized cyclodextrins that are able to perform many
enantiomeric separations.
Chiral Test Mix for Astec CHIROBIOTIC®
5-Methyl-5-phenylhydantoin
C10H10N2O2 FW 190.20 Related Information
 analytical standard
5-Methyl-5-phenylhydantoin is used to evaluate the performance of Astec
CHIROBIOTIC® chiral HPLC columns. The mobile phase is 100% methanol No. Title
T411101 Supelco Chiral GC Columns
and detection is by UV at 254 nm. The test mix is supplied as a racemic
mixture of two enantiomers in methanol.
Components
5-Methyl-5-phenylhydantoin 5000 μg/mL Cyclodextrin-based GC CSPs
40095-U 1 mL
Cyclodextrins (CDs) are macromolecules composed of 6 or more D
(+)-glucose residues bonded through α-glycosidic linkages. They are
Chiral Normal Phase Test Mix classified according to the number of glucose residues they contain: α-
 30 μg/mL each component in hexane, analytical standard cyclodextrins (six residues), β-cyclodextrins (seven residues), and γ-cyclo-
dextrins (eight residues). The three different sizes separate analytes over a
Trans-stilbene oxide (TSO) is used to evaluate the performance of Astec
wide range of molecular size. All hydroxyl groups, whether at the 2, 3 or 6
Cellulose DMP and other polysaccharide-based chiral HPLC columns. The
position, can be selectively modified with a derivative to impart unique
recommended mobile phase is 10:90 IPA:hexane and detection is by UV at
physical properties and inclusion selectivities. Unlike LC, there is no
220 nm. The test mix is supplied as a racemic mixture of the two TSO
enantioselectivity in chiral GC without derivatization of the CD.
enantiomers with 1,3,5-tri-tert-butylbenzene as a void volume marker. The
solvent is hexane.
Components
trans-Stilbene oxide
1,3,5-tri-t-Butylbenzene
40119-U 1 mL
Cyclodextrin molecules showing dimensions

Chiral GC Columns
Cyclodextrin-based GC CSPs
Cyclodextrins—Physical Properties Group 3: Inclusion Interactions

Glucose Stereogenic The third group of GC CSPs relies on inclusion interactions for retention. The
Cyclodextrin Units Centers MW Cavity (nm) fact that there are three different size cyclodextrins (α, β and γ) allows for
Alpha 6 30 972 0.57 separation of a wide variety of different size analytes. This group includes
Beta 7 35 1135 0.78 CHIRALDEX® DM and CHIRALDEX® DA, and Supelco DEX 110 and Supelco
Gamma 8 40 1297 0.95 DEX 120.
CD Derivatives in the Supelco Chiral GC Line:

In cyclodextrin-based capillary GC CSPs, the derivatized cyclodextrin is used
neat or after being doped at controlled percentages into a polysiloxane Cyclodextrin Type
polymer matrix. Cyclodextrin GC CSPs are grouped into three general Derivative Phase α (alpha) ß (beta) γ (gamma)
categories: Butyryl Astec CHIRALDEX BP ✓
Diacetyl Supelco DEX 225 ✓ ✓ ✓
Group 1: Surface Interactions, Complex Derivatives
Because the predominant mechanism of retention for phases in this group Dialkyl Astec CHIRALDEX DA ✓ ✓ ✓
is based on surface interaction, γ-cyclodextrin, with eight glucose molecules, Dimethyl Astec CHIRALDEX DM ✓ ✓
has been shown to be the most useful. Compared to α− and Dimethyl Supelco DEX 325 ✓ ✓ ✓
β−cyclodextrins, the greater number of glucose molecules in a γ-cyclo- Dipropionyl Astec CHIRALDEX DP ✓ ✓
dextrin results in the greater number of 2,3,6-position hydroxyl functional S-Hydroxypropyl Astec CHIRALDEX PH ✓
groups available for derivatization. High derivative concentration is Permethylated Astec CHIRALDEX PM ✓
beneficial for maximizing surface interactions. This group includes the highly Permethylated Supelco DEX 110 ✓
popular CHIRALDEX® G-TA. Permethylated Supelco DEX 120 ✓ ✓ ✓
Group 2: Surface/Inclusion Interactions, Simple Derivatives Propionyl Astec CHIRALDEX PN ✓
This group includes the diacetyl (Supelco DEX 225) and dimethyl (Supelco Trifluoroacetyl Astec CHIRALDEX TA ✓ ✓ ✓
DEX 325) derivatives. The β-cyclodextrin has shown the greatest applic-
ability for phases with these derivatives.
Choosing a Chiral GC Column

Supelco offers the most extensive line of chiral capillary GC columns in the industry. Our two premium lines of GC CSPs, Supelco DEX and Astec CHIRALDEX®,
comprise a wide range of cyclodextrin derivatives with complementary selectivity. All are stable, high boiling liquids and make effective CSPs for enantiomer
separations by GC. Selectivity is a function of the derivative, the degree of derivatization, the position of the derivative on the cyclodextrin, whether the
derivatized cyclodextrin is used neat or doped into a polysiloxane, and if doped, at what percentage. Certain CSPs are more selective for given molecular
structures and often more than one will achieve a separation. CSPs may be chosen to optimize resolution, but also elution order or analysis time. It is
conventional practice to screen multiple CSPs when developing a new method. We offer column screening kits at very attractive prices for this purpose.
Supelco Chiral Capillary GC Column Selection Guidelines

Supelco DEX 325
Supelco DEX 225
Supelco DEX 120
Supelco DEX 110

CHIRALDEX DM
CHIRALDEX PM
CHIRALDEX DA
CHIRALDEX DP
CHIRALDEX PN
CHIRALDEX PH
CHIRALDEX TA
CHIRALDEX BP
β-Cyclodextrin
α-Cyclodextrin
γ-Cyclodextrin
——————————————— By Chemistry —————————————— – By Cyclodextrin –
Oxygen containing analytes in the form of alcohols, ✓
ketones, acids, aldehydes, and lactones; halogenated
compounds
Aliphatic and aromatic amines; aliphatic and some ✓
aromatic esters; polar racemates
Lactones and aromatic amines; epoxides; styrene oxide ✓
Amino acids; amines; furans ✓
Aliphatic, olefenic, and aromatic enantiomers ✓ ✓ ✓ ✓
Terpenes and tertiary amines ✓ ✓ ✓
Heterocyclic amines ✓
Xylenes, menthols, cresols, substituted phenols, substi- ✓
tuted benzenes, epoxide enantiomers
Acids, alcohols, amines, diols, esters, ethers, halohydro- ✓
carbons, hydrocarbons, ketones, positional isomers, silanes,
terpenes, terpineols
α-BHC, carvone, carboxylic acids, methamphetamine ✓

Chiral GC Columns
Chiral GC Column Screening Kits
cyclodextrin results in the greater number of 2,3,6-position hydroxyl

Chiral GC Column Screening Kits functional groups available for derivatization. High derivative concentration
These column screening kits provide the necessary columns to perform is beneficial for maximizing surface interactions.
most chiral separations and run mechanistic studies, and are offered at very
attractive prices. Astec CHIRALDEX® G-TA is the first choice in this group. This phase has
been shown to be the most broadly selective phase for the pharmaceutical
industry, especially in the analysis of chiral intermediates and drug studies in
Related Information various stages of clinical trials. Separations occur without the inclusion
Need help choosing the right chiral HPLC or GC column? mechanism and are typically faster and more efficient than most chiral
Let our Chiral Services group do the work for you. stationary phases. This phase does not contain a polysiloxane carrier and,
therefore, there are no deleterious effects at low temperatures. The ability of
this phase to separate parent drug enantiomers and their metabolites has
proven quite beneficial. A modified version of the G-TA is the Astec
Astec CHIRALDEX® GC Column Screening Kit CHIRALDEX® G-PN. It functions like the G-TA but shows higher selectivity
toward certain amines (amphetamine, methamphetamine). This phase is
The Astec CHIRALDEX® column kit contains three GC CSPs that cover the more stable to moisture than the G-TA.
widest possible range of enantioselectivity: CHIRALDEX® G-TA, B-DM, and B-
The Astec CHIRALDEX® G-DP phase was introduced to enhance selectivity
DA, in the popular 30 m x 0.25 mm, 0.12 μm df dimensions. The CHIRALDEX®
for both aliphatic and aromatic amines in additional to aliphatic and some
G-TA separates the greatest number of enantiomers, often with high
aromatic esters. This phase is especially useful for polar racemates. This
enantioselectivity. The CHIRALDEX® B-DM separates the widest variety of
phase demonstrates better hydrolytic and thermal stability than the G-TA.
different structural types. The CHIRALDEX® B-DA is best suited for larger
The Astec CHIRALDEX® G-BP phase can be used as a general purpose
multi-ring structures. Eighty-five percent of analytes that exhibit enantiose-
column but it is especially useful for amino acids.
lectivity on cyclodextrin based chiral stationary phases will give enantiose-
lectivity on one of these phases. The kit provides considerable savings over
Note: The subtle differences in functional groups between the G-TA, G-DP, G-PN,
the columns purchased separately.
and G-BP often allow for major enhancements in chiral and achiral selectivity
Kit contents: One 30 m × 0.25 mm I.D., 0.12 μm column of each type: when changing from one phase to another.
CHIRALDEX® G-TA, B-DM, and B-DA
Astec CHIRALDEX® GC Column Screening Kit Trifluoroacetyl (TA) Cyclodextrin Derivatives

Astec CHIRALDEX® A-TA, B-TA, and G-TA
Description Cat. No. Qty
30 m kit 71030AST 1 kit Trifluoroacetylation of the 3-hydroxyl group after pentylation of the 2,6-
hydroxyl groups creates a phase with high selectivity for oxygen-containing
analytes in the form of alcohols, ketones, acids, aldehydes, lactones. Highly
selective for halogenated compounds. Astec CHIRALDEX® G-TA is the most
Supelco DEX™ GC Column Screening Kit popular phase in our chiral GC line.
These Supelco DEX kits provide the tools you need to perform most chiral Features
separations. Confirm identities of enantiomers by monitoring elution order • Phase: 2,6-di-O-pentyl-3-trifluoroacetyl derivative of α-, β-, or γ-cyclo-
changes (enantioreversal) from one column to another. In combination, the dextrin
columns in the two kits span the full range of Supelco DEX column • Separates the widest variety and greatest number of enantiomers
enantioselectivity. Compare the savings to the columns purchased • Unique retention behavior
separately. • Extraordinary versatility and chiral selectivity
Kit I: One 30 m × 0.25 mm I.D., 0.25 μm column of each type: α-DEX 120, β- • Sensitive to moisture, but can be regenerated
DEX 120 and γ-DEX 120 • Thermal limit 180 °C (isothermal or programmed)
Kit II: One 30 m × 0.25 mm I.D., 0.25 μm column of each type: β-DEX 120, β-
Analytes
DEX 225, γ-DEX 225 and β-DEX 325
• Useful for separating homologous series of amino acids (primary,
Supelco DEX™ GC Column Screening Kit secondary, aromatic and aliphatic), amines (primary, secondary, cyclic,
aromatic and halogenated), amino alcohols, alkanes, hydrogenated
Description Cat. No. Qty alkanes, alcohols (aliphatic and aromatic), acids (halogenated and
kit I 24340 1 kit hydroxy), esters (aromatic, aliphatic, hydroxy, di-ester), diols, lactones,
kit II 24328-U 1 kit ketones, phthalides, and sulphoxides
Mechanism Observations
• Strong dipole-dipole interactions
Group 1: Surface Interactions, Complex • Longer alkyl chain; greater retention; increase in enantioselectivity up to
Derivatives C4/C5
• Halogens known to favor cavity interaction
Sigma-Aldrich is the only supplier of complex derivatives for chiral GC. There • Dipole-dipole interactions are commonly identified in the mechanism of
are four members in this important group: separation for CHIRALDEX® TA phases. In a homologous series of alkane
• Astec CHIRALDEX® TA (Trifluoroacetyl derivatives) enantiomers, identical alpha values are observed regardless of chain
• Astec CHIRALDEX® PN (Propionyl derivatives) length or branching indicating only 1 or 2 carbons may be contributing to
• Astec CHIRALDEX® DP (Dipropionyl derivatives) chiral recognition. Alpha values are greatly affected by size and polarity of
• Astec CHIRALDEX® BP (Butyryl derivatives) the head group. Functional groups like epoxides, amino alcohols and
alcohols can dictate the cyclodextrin selection. Aldehydes, carboxylic acids
Because the predominant mechanism of retention for phases in this group and epoxides separate better on the gamma while alcohols, alcohol
is based on surface interaction, the gamma-cyclodextrin, with 8 glucose amines and other linear molecules separate better on the beta derivative.
molecules, has been shown to be the most useful. Compared to alpha- and
beta-cyclodextrins, the greater number of glucose molecules in a gamma-

Chiral GC Columns
Group 1: Surface Interactions, Complex Derivatives: Trifluoroacetyl (TA) Cyclodextrin Derivatives
Size Selectivity
Astec CHIRALDEX® TA Series Column Regeneration
• The γ−TA (CHIRALDEX G-TA) derivative has proven to exhibit a wider chiral
selectivity and usefulness than the β (CHIRALDEX® B-TA) analog. The
Reagents
influence of the inclusion mechanism for chiral recognition is very much The trifluoroacetic anhydride (TFA) derivative of the cyclodextrins used in
reduced and capacities are generally higher indicating more surface CHIRALDEX® TA phases can hydrolyze in the presence of moisture at and
interaction. Of all the compounds we have tested, the split between above room temperature. Sources of moisture include the sample, carrier
CHIRALDEX® G-TA and CHIRALDEX® B-TA is approximately 55/35 with only gas, injection solvents and the atmosphere if stored unsealed. For long
10% of the separations accomplished on the α (CHIRALDEX® A-TA). column life, ensure that the carrier gas line has an effective moisture trap, all
sample extracts are moisture free, the injection solvent is anhydrous and the
Astec CHIRALDEX® A-TA Capillary GC Column column is stored properly when not in use by flame-sealing the ends. Guard
columns will also help protect the column from the damaging effects of
Incorporates a phase consisting of a 2,6-di-O-pentyl-3-trifluoroacetyl moisture. This regeneration method does not restore retention due to loss
derivative of α-cyclodextrin. This phase exhibits high selectivity for oxygen- of phase, it restores enantioselectivity lost by hydrolysis of the acetyl
containing analytes in the form of alcohols, ketones, acids, aldehydes and derivative. Instructions for regeneration of CHIRALDEX® TA columns are
lactones. It is also highly selective for halogenated compounds. available by contacting [email protected].
Temp. Limits:
For flame-sealing the ends of the column, we offer the Microflame Gas
• -10 °C to 180 °C, isothermal and programmed Torch Set (Cat. No. 22969). Details of this item can be found in the
phase ....................................... non-bonded; 2,6-di-O-pentyl-3-trifluoroacetyl derivative of α-cyclodextrin Laboratory Supplies section of this catalog.
I.D. Length Beta Description Cat. No. Qty
(mm) df (μm) (m) Value Cat. No. Qty
Trifluoroacetic anhydride, for GC derivati- 33164 25 mL
0.25 0.12 20 500 73002AST 1 ea zation
0.12 30 500 73003AST 1 ea Trifluoroacetic anhydride 33165-U 10 × 1 mL
0.12 40 500 73004AST 1 ea 33164 25 mL
0.12 50 500 73005AST 1 ea Methyl Red, ACS reagent, crystalline 250198-25G 25 g
250198-100G 100 g
Astec CHIRALDEX® B-TA Capillary GC Column Sodium hydroxide, ACS reagent, ≥97.0%, 221465-25G 25 g
pellets 221465-500G 500 g
Incorporates a phase consisting of a 2,6-di-O-pentyl-3-trifluoroacetyl 221465- 6 × 500 g
6X500G 2.5 kg
derivative of β-cyclodextrin. This phase exhibits high selectivity for oxygen- 221465- 12 kg
containing analytes in the form of alcohols, ketones, acids, aldehydes and 2.5KG 50 kg
lactones. It is also highly selective for halogenated compounds. 221465-12KG
Temp. Limits: 221465-50KG
• -10 °C to 180 °C isothermal and programmed

phase ....................................... non-bonded; 2,6-di-O-pentyl-3-trifluoroacetyl derivative of β-cyclodextrin Propionyl (PN) Cyclodextrin Derivatives
I.D. Length Beta Astec CHIRALDEX® G-PN
0.25 0.12 20 500 73022AST 1 ea A modified version of the CHIRALDEX® G-TA, Astec CHIRALDEX® G-PN
0.12 30 500 73023AST 1 ea exhibits high selectivity for lactones, epoxides, and aromatic amines.
0.12 40 500 73024AST 1 ea Additionally, the analysis of styrene oxide can be accomplished on this
phase (this analyte degrades on the TA phases). This phase is more stable to
Astec CHIRALDEX® G-TA Capillary GC Column moisture than the CHIRALDEX® G-TA.
Features
Astec CHIRALDEX G-TA is the first choice in the Group 1 CSPs (Surface
Interactions, Complex Derivatives). This phase has been shown to be the • Phase: 2,6-di-O-pentyl-3-propionyl derivative of γ-cyclodextrin
most broadly-selective phase for the pharmaceutical industry, especially for • Suitable for epoxide separations
the analysis of chiral intermediates and drug studies in various stages of • High selectivity for lactones
clinical trials. Separations occur without the inclusion mechanism and are • High selectivity for aromatic amines (i.e. amphetamine, methamphet-
typically faster and more efficient than most chiral stationary phases. G-TA amine)
has also been used to separate parent drug enantiomers and their • More stable to moisture than the CHIRALDEX® TA phases
metabolites. G-TA has its highest selectivity for oxygen-containing analytes • Thermal limit 200/220°C (isothermal/programmed)
like alcohols, diols and polyols as the free alcohol and as an acyl derivative; Analytes
amines as acyl derivatives; amino alcohols, halogens (Cl>Br>F), amino acids,
hydroxy acids, lactones, furans and pyrans. It is also highly selective for • Epoxides, aromatic amines (amphetamine/methamphetamine), >C6
halogenated compounds. alcohols and lactones
Temp. Limits: Mechanism Observations
• -10 °C to 180 °C isothermal and programmed • There is little evidence of inclusion formation. Retention increases with
phase ........................................ non-bonded; 2,6-di-O-pentyl-3-trifluoroacetyl derivative of γ-cyclodextrin increased chain length of analyte. This allows for efficient separation of a
I.D. Length Beta
series of homologs.
Size Selectivity
0.25 0.12 10 500 73031AST 1 ea
• The CHIRALDEX® G-PN shows very little size selectivity.
0.12 20 500 73032AST 1 ea
0.12 30 500 73033AST 1 ea
0.12 40 500 73034AST 1 ea
0.12 50 500 73035AST 1 ea

Chiral GC Columns
Group 1: Surface Interactions, Complex Derivatives: Propionyl (PN) Cyclodextrin Derivatives
8P R O D U C T S 8P R O D U C T S
Astec CHIRALDEX® G-PN Capillary GC Column Astec CHIRALDEX® B-DP Capillary GC Column
Incorporates a phase consisting of a 2,6-di-O-pentyl-3-propionyl derivative Incorporates a phase consisting of a 2,3-di-O-propionyl-6-t-butyl silyl
of γ-cyclodextrin. This phase exhibits high selectivity for lactones and derivative of β-cyclodextrin. This phase exhibits good hydrolytic stability,
aromatic amines. It is also suitable for epoxide separations. Additionally, the broad chiral selectivity, and is excellent for aliphatic and aromatic amines. It
analysis of styrene oxide can be accomplished on this phase (this analyte is also good for many aliphatic and some aromatic esters as well as
degrades on the TA phases). exhibiting high efficiency and resolution at low retention times for polar
GC capillary column racemates.
fused silica Temp. Limits:
Temp. Limits: • -10 °C to 200 °C isothermal, 220 °C programmed
• -10 °C to 200 °C isothermal, 220 °C programmed phase ...................................... non-bonded; 2,3-di-O-propionyl-6-t-butyl silyl derivative of β-cyclodextrin
phase .................................................. non-bonded; 2,6-di-O-pentyl-3-propionyl derivative of γ-cyclodextrin I.D. Length Beta
I.D. Length Beta
(mm) df (μm) (m) Value Cat. No. Qty 0.25 0.12 30 500 78023AST 1 ea
0.25 0.12 30 500 74033AST 1 ea
Astec CHIRALDEX® G-DP Capillary GC Column
Incorporates a phase consisting of a 2,3-di-O-propionyl-6-t-butyl silyl
Dipropionyl (DP) Cyclodextrin Derivatives derivative of γ-cyclodextrin. The CHIRALDEX G-DP phase was designed to
Astec CHIRALDEX® B-DP and G-DP enhance selectivity for both aliphatic and aromatic amines, in additional to
aliphatic and some aromatic esters. This phase is especially useful for polar
This derivative demonstrates good selectivity for a wide range of analytes
racemates, as it exhibits high efficiency and resolution at low retention
except alcohols and epoxides where the CHIRALDEX® G-TA remains the best
times. G-DP demonstrates better hydrolytic and thermal stability than the
choice. The CHIRALDEX® G-DP has shown very high selectivity for both
CHIRALDEX G-TA.
aromatic and aliphatic amines and for aliphatic and some aromatic esters.
Both hydrolytic and temperature stability are better than CHIRALDEX® G-TA, Temp. Limits:
and for bulky fused ring structures the CHIRALDEX® G-DP is better than the • -10 °C to 200 °C isothermal, 220 °C programmed
CHIRALDEX® B-DP. phase ....................................... non-bonded; 2,3-di-O-propionyl-6-t-butyl silyl derivative of γ-cyclodextrin
Features I.D. Length Beta
• Phase: 2,3-di-O-propionyl-6-t-butyl silyl derivative of β- or γ-cyclodextrin
0.25 0.12 30 500 78033AST 1 ea
• Broad chiral selectivity
• Good hydrolytic stability
• High efficiency and resolution at low retention times for polar racemates
• Thermal limit 200/220 °C (isothermal/programmed)
Butyryl (BP) Cyclodextrin Derivatives
Astec CHIRALDEX® G-BP
Astec CHIRALDEX® G-BP incorporates a phase consisting of a 2,6-di-O-
• Mostly surface interactions
pentyl-3-butyryl derivative of γ-cyclodextrin. It is a good general-purpose
• Fused ring structures better selectivity on gamma
column. It is also especially useful for amino acids and is, therefore, a good
• Acids have better selectivity as methyl rather than ethyl esters
substitute for the bonded amino acid-type chiral GC phases.
Analytes Features
• Excellent for aromatic and aliphatic amines • Phase: 2,6-di-O-pentyl-3-butyryl derivative of γ-cyclodextrin
• Good for many aliphatic and some aromatic esters • High selectivity for amino acids, amines, and furans
Size Selectivity • High sample capacity
• Speed and sample capacity point to a surface-type mechanism for very
polar racemates. Large, bulky molecules still require a larger surface area Mechanism Observations
than the beta provides, therefore, an increase in selectivity is seen on the • Alkyl chain on analyte contributes to chiral recognition
gamma derivative for fused ring structures. The smaller alpha cavity • High sample capacity indicates primarily surface interactions
offered no selectivity while the beta covered the largest range of
molecular sizes. The choice between beta and gamma is compound Analytes
dependent for this phase. • Amino acids, amines and furans
Size Selectivity
• The influence of the inclusion mechanism on selectivity is much reduced
and capacities are, therefore, generally higher.

Chiral GC Columns
Group 1: Surface Interactions, Complex Derivatives: Butyryl (BP) Cyclodextrin Derivatives
8P R O D U C T S
α-DEX™ 225
Astec CHIRALDEX® G-BP Capillary GC Column The chiral stationary phase in α-DEX 225 columns contains 2,3-di-O-acetyl-6-
0-TBDMS-α-cyclodextrin embedded in an intermediate polarity phase.
Incorporates a phase consisting of a 2,6-di-O-pentyl-3-butyryl derivative of γ-
cyclodextrin. This phase exhibits high selectivity for amino acids, amines, Temp. Limits:
and furans. • 30 °C to 230 °C
GC capillary column phase ...................................................................... non-bonded; 25% 2,3-di-O-acetyl-6-O-TBDMS-α-cyclodextrin
fused silica in SPB-20 poly(20% phenyl/80% dimethylsiloxane)
Temp. Limits: I.D. Length Beta

• -10 °C to 200 °C isothermal, 220 °C programmed 0.25 0.25 30 250 24311 1 ea
phase ........................................................ non-bonded; 2,6-di-O-pentyl-3-butyryl derivative of γ-cyclodextrin
I.D. Length Beta β-DEX™ 225
0.25 0.12 30 500 75033AST 1 ea The Supelco β-DEX 225 is a modified form of the β-DEX 325 phase, and
employs acetyl derivatives at the 2,3-positions instead of more traditional
methyl derivatives. The chiral stationary phase in β-DEX 225 columns
contains 2,3-di-O-acetyl-6-0-TBDMS-β-cyclodextrin embedded in an inter-
Group 2: Surface/Inclusion Interactions, mediate polarity phase. These columns provide unique selectivity for
enantiomeric separations of small molecules: alcohols, aldehydes (e.g., 2-
Simple Derivatives phenylpropionaldehyde), esters (e.g. methyl malate, methyl lactate), flavor
There are three different derivatives in this group: compounds and ketones.
• Supelco DEX™ 225 (Diacetyl derivatives) Temp. Limits:
• Astec CHIRALDEX® DM and Supelco DEX™ 325 (Dimethyl derivatives) • 30 °C to 230 °C
• Astec CHIRALDEX® PM, Supelco DEX™ 110, and DEX™ 120 (Permethyl phase ................................................................ non-bonded; 25% 2,3-di-O-acetyl-6-O-TBDMS-β-cyclodextrin in
derivatives) SPB-20 poly(20% phenyl/80% dimethylsiloxane)
The beta-cyclodextrin has shown the greatest applicability for phases with I.D. Length Beta
these derivatives. Astec CHIRALDEX® B-DM is the recommended column in
0.25 0.25 30 250 24348 1 ea
this category. The Supelco β-DEX 325 is similar in both chemistry and use to
the CHIRALDEX® B-DM phase, the main difference being the concentration
of the dimethyl-derivatized cyclodextrin that is doped into the polysiloxane γ-DEX™ 225
carrier. The Supelco β-DEX 225 is a modified form of the β-DEX 325 phase, The chiral stationary phase in γ-DEX 225 columns contains 2,3-di-O-acetyl-6-
employing acetyl derivatives at the 2,3-positions instead of more traditional 0-TBDMS-γ-cyclodextrin embedded in an intermediate polarity phase.
methyl derivatives. Temp. Limits:
This group also includes the popular dimethyl and permethyl derivatives, • 30 °C to 230 °C
and includes Astec CHIRALDEX® B-PM, Supelco β-DEX 110, and Supelco β- phase ....................................................................... non-bonded; 25% 2,3-di-O-acetyl-6-O-TBDMS-γ-cyclodextrin
DEX 120 phases. They are recommended as general purpose columns for in SPB-20 poly(20% phenyl/80% dimethylsiloxane)
the separation of a wide variety of compounds and are especially useful for I.D. Length Beta
the analysis of alcohols and diols in their underivatized form and analytes (mm) df (μm) (m) Value Cat. No. Qty
with polar groups (such as tertiary amines).The main difference between 0.25 0.25 30 250 24312 1 ea
these three phases is the concentration of the permethyl-derivatized
cyclodextrin that is doped into the polysiloxane carrier.
Dimethyl (DM, 325) Cyclodextrin Derivatives
Diacetyl (225) Cyclodextrin Derivatives Astec CHIRALDEX® B-DM and G-DM
Supelco DEX™ 225 The dimethyl derivative was designed to overlap with the applications of
The Supelco DEX 225 phases are modified forms of the DEX 325 phases, both Astec CHIRALDEX® PM and CHIRALDEX® PH series, having similar
employing acetyl derivatives at the 2,3-positions instead of more traditional selectivity but with shorter retention times and better resolution.
methyl derivatives. The chiral stationary phase in DEX 225 columns contains Features
2,3-di-O-acetyl-6-0-TBDMS derivatized α, β, or γ cyclodextrin embedded in
• Phase: 2,3-di-O-methyl-6-t-butyl silyl derivative of β- or γ-cyclodextrin
an intermediate polarity phase. These columns provide unique selectivity for
• Broad chiral selectivity
enantiomeric separations of small molecules: alcohols, aldehydes (e.g., 2-
• Combines selectivity of CHIRALDEX® PM (permethyl) and CHIRALDEX® PH
phenylpropionaldehyde), esters (e.g., methyl malate, methyl lactate), flavor
(hydroxypropyl)
compounds and ketones.
• Short retention, high resolution
• β derivative is broadly applicable
Features
• Phase: 25% 2,3-di-O-acetyl-6-O-TBDMS-α-, β,- or γ-cyclodextrin in SPB-20
poly(20% phenyl/80% dimethylsiloxane) Analytes
• Thermal limit 230 °C (isothermal or programmed) • Resolves aliphatic, olefinic and aromatic enantiomers
Analytes Mechanism Observations
• Alcohols, aldehydes (e.g., 2-phenylpropionaldehyde), esters (e.g., methyl • Size selectivity present but not dominant as in CHIRALDEX® DA
malate, methyl lactate), flavor compounds, ketones • Fewer structural requirements
• Characteristic temperature selectivity

Chiral GC Columns
Group 2: Surface/Inclusion Interactions, Simple Derivatives: Dimethyl (DM, 325) Cyclodextrin Derivatives
Size Selectivity β-DEX™ 325

• Size selectivity is evident, implying that the inclusion mechanism plays a The chiral stationary phase in β-DEX 325 columns contains 2,3-di-O-methyl-
role in the separation mechanism, but does not dominate as it does in the 6-0-TBDMS-β-cyclodextrin embedded in an intermediate polarity phase. The
CHIRALDEX® DA series. The β form covers a very broad range of molecular Supelco β-DEX 325 is similar in both chemistry and use to the CHIRALDEX B-
sizes and, therefore, has the widest applicability. DM phase, the main difference being the concentration of the dimethyl-
Supelco DEX™ 325 derivatized cyclodextrin that is doped into the polysiloxane carrier.
Temp. Limits:
The chiral stationary phase in DEX 325 columns is the 2,3-di-O-methyl-6-0-
TBDMS derivative of α-, β-, or γ-cyclodextrin embedded in an intermediate • 30 °C to 230 °C
polarity phase. The Supelco ß-DEX 325 is similar in both chemistry and use phase .................................................................... non-bonded; 25% 2,3-di-O-methyl-6-O-TBDMS-β-cyclodextrin
to the Astec CHIRALDEX® B-DM phase; the main difference being the in SPB-20 poly(20% phenyl/80% dimethylsiloxane)
concentration of the dimethyl-derivatized cyclodextrin that is doped into I.D. Length Beta
the polysiloxane carrier. (mm) df (μm) (m) Value Cat. No. Qty
0.25 0.25 30 250 24308 1 ea
Features
• Phase: 25% 2,3-di-O-methyl-6-O-TBDMS-α-, β-, or γ-cyclodextrin in SPB-20 γ-DEX™ 325
poly(20% phenyl/80% dimethylsiloxane) The chiral stationary phase in Supelco γ-DEX 325 columns contains 2,3-di-O-
• Thermal limit 230 °C (isothermal or programmed) methyl-6-0-TBDMS-γ-cyclodextrin embedded in an intermediate polarity
phase.
8P R O D U C T S Temp. Limits:
• 30 °C to 230 °C
Astec CHIRALDEX® B-DM Capillary GC Column phase .................................................................... non-bonded; 25% 2,3-di-O-methyl-6-O-TBDMS-γ-cyclodextrin
in SPB-20 poly(20% phenyl/80% dimethylsiloxane)
Through special derivatization techniques, the concentration of the
cyclodextrin in the CHIRALDEX B-DM has been substantially increased in the I.D. Length Beta
polysiloxane carrier. This phase is very useful for a number of free acids and
0.25 0.25 30 250 24306 1 ea
bases. The B-DM is able to perform most of the separations done on a beta-
permethylated phase, but with higher resolution. The selectivity of the B-DM
covers applications of both the B-PM and B-PH phases, although with Selection kits offer the greatest likelihood of
superior performance. providing the separation you want
Temp. Limits:
Terpinen-4-ol
• -10 °C to 200 °C isothermal, 220 °C programmed (+)
phase ............................................ non-bonded; 2,3-di-O-methyl-6-t-butyl silyl derivative of β-cyclodextrin b-DEX 325
(+)
I.D. Length Beta
0.25 0.12 20 500 77022AST 1 ea
(–) b -DEX 225
0.12 30 500 77023AST 1 ea
0.12 40 500 77024AST 1 ea
0.12 50 500 77025AST 1 ea
Astec CHIRALDEX® G-DM Capillary GC Column

Incorporates a phase consisting of a 2,3-di-O-methyl-6-t-butyl silyl derivative
of γ-cyclodextrin. This phase exhibits broad chiral selectivity, resolving
aliphatic, olefenic, and aromatic enantiomers. It combines the selectivities of
the PM and PH phases. (–)
Temp. Limits:
• -10 °C to 200 °C isothermal, 220 °C programmed
phase ............................................. non-bonded; 2,3-di-O-methyl-6-t-butyl silyl derivative of γ-cyclodextrin
20 Min 30 Min
I.D. Length Beta
0.25 0.12 30 500 77033AST 1 ea
α-DEX™ 325
The chiral stationary phase in α-DEX 325 columns contains 2,3-di-O-methyl-
6-0-TBDMS-α-cyclodextrin embedded in an intermediate polarity phase.
Temp. Limits:
• 30 °C to 230 °C
phase .................................................................... non-bonded; 25% 2,3-di-O-methyl-6-O-TBDMS-α-cyclodextrin
in SPB-20 poly(20% phenyl/80% dimethylsiloxane)
I.D. Length Beta
0.25 0.25 30 250 24303 1 ea

Chiral GC Columns
Group 2: Surface/Inclusion Interactions, Simple Derivatives: Permethyl (PM, 110, 120) Cyclodextrin Derivatives
Permethyl (PM, 110, 120) Cyclodextrin Derivatives 8P R O D U C T S

The main difference between Astec CHIRALDEX B-PM and Supelco β-DEX
110 and Supelco β-DEX 120 is the concentration of the permethyl- Astec CHIRALDEX® B-PM Capillary GC Column
derivatized cyclodextrin that is doped into the polysiloxane carrier. This The main difference between CHIRALDEX B-PM and the Supelco β-DEX 110
confers selectivity differences between the columns. and Supelco β-DEX 120 phases is the concentration of the permethyl-
Astec CHIRALDEX® B-PM derivatized cyclodextrin that is doped into the polysiloxane carrier.
CHIRALDEX B-PM is a general-purpose column used for the separation of
The permethylated β-cyclodextrin is a GC CSP with potential to separate a
acids, alcohols, barbitals, diols, epoxides, esters, hydrocarbons, ketones,
wide variety of racemates. The PM is especially valuable for separating
lactones and terpenes. Also, some underivatized alcohols and diols as well
certain hydrocarbons, like terpenes, some underivatized alcohols and diols,
as some analytes with polar groups, i.e. tertiary amines, show excellent
and some analytes with polar groups like tertiary amines.
separation.
Features Temp. Limits:
• Phase: 2,3,6-tri-O-methyl derivative of β-cyclodextrin • -10 °C to 200 °C isothermal, 220 °C programmed
• Broad chiral selectivity phase ......................................................................... non-bonded; 2,3,6-tri-O-methyl derivative of β-cyclodextrin
• Strong inclusion/size selectivity
I.D. Length Beta
• Thermal limit 200/220 °C (isothermal/programmed) (mm) df (μm) (m) Value Cat. No. Qty
Analytes 0.25 0.12 30 500 76023AST 1 ea
• A general purpose column used for the separation of acids, alcohols, 0.12 50 500 76025AST 1 ea
barbitals, diols, epoxides, esters, hydrocarbons, ketones, lactones, and
terpenes. β-DEX™ 110
• Some underivatized alcohols and diols as well as some analytes with polar The chiral stationary phase in β-DEX 110 columns contains permethylated β-
groups, e.g. tertiary amines, show excellent separation. cyclodextrin embedded in an intermediate polarity stationary phase. They
Mechanism Observations are recommended for the enantiomeric separation of a wide range of chiral
compounds (ketones, esters, alkanes, alkenes, alcohols, acids, ethers, etc.).
• Inclusion a dominant mechanism The 10% (β-DEX 110) and 20% (β-DEX 120) β-cyclodextrin content alters the
• Highest temperature stability of CHIRALDEX phases along with the elution order while maintaining similar enantioselectivity.
CHIRALDEX DM
Temp. Limits:
Size Selectivity • 30 °C to 230 °C
• Some size selectivity is evident with the permethylated phase. The beta phase ............... non-bonded; 10% permethylated β-cyclodextrin in SPB-35 poly(35% diphenyl/65%
form will cover a broad range of molecule sizes. dimethylsiloxane)
Supelco DEX™ 110 and 120 I.D. Length Beta

The chiral stationary phases in DEX 110 and 120 columns contain 0.25 0.25 30 250 24301 1 ea
permethylated cyclodextrins embedded in an intermediate polarity 0.25 60 250 24302 1 ea
stationary phase. These columns are recommended for the enantiomeric
separation of a wide range of chiral compounds (ketones, esters, alkanes, α-DEX™ 120
alkenes, alcohols, acids, ethers, etc.). The 10% (β-DEX 110) and 20% (β-DEX
120) β-cyclodextrin content alters the elution order while maintaining Containing permethylated α-cyclodextrin embedded in an intermediate
similar enantioselectivity. polarity stationary phase, Supelco α-DEX 120 columns provide unique
selectivity for enantiomeric separations of small molecules. They are also
Because the elution order of a pair of enantiomers frequently reverses recommended for separating positional isomers (phenols, xylenes, etc.).
(enantioreversal) on a γ-DEX column compared to the elution order on an α- Temp. Limits:
DEX or β-DEX column, we recommend γ-DEX 120 columns as complements • 30 °C to 230 °C
to α-DEX 120 and β-DEX 120 columns. The γ-DEX is useful for enantiomeric
phase ............... non-bonded; 20% permethylated α-cyclodextrin in SPB-35 poly(35% diphenyl/65%
differentiation of large analytes, e.g. α-BHC, carvone, carboxylic acids, and dimethylsiloxane)
methamphetamine.
I.D. Length Beta
Features (mm) df (μm) (m) Value Cat. No. Qty
0.25 0.25 30 250 24310 1 ea
• Phase (110): 10% permethylated β-cyclodextrin in SPB-35 poly(35%
diphenyl/65% dimethylsiloxane)
• Phase (120): 20% permethylated α-, β-, or γ-cyclodextrin in SPB-35 poly β-DEX™ 120
(35% diphenyl/65% dimethylsiloxane) The chiral stationary phase in β-DEX 120 columns contains permethylated β-
• Thermal limit 230 °C (isothermal or programmed) cyclodextrin embedded in an intermediate polarity stationary phase. They
are recommended for the enantiomeric separation of a wide range of chiral
compounds (ketones, esters, alkanes, alkenes, alcohols, acids, ethers, etc.).
The 10% (β-DEX 110) and 20% (β-DEX 120) β-cyclodextrin content alters the
elution order while maintaining similar enantioselectivity.
Temp. Limits:
• 30 °C to 230 °C
phase ................... non-bonded; 20% permethylated β-cyclodextrin in SPB-35 poly(35% phenyl/65%
dimethylsiloxane)
I.D. Length Beta
0.25 0.25 30 250 24304 1 ea
0.25 60 250 24305-U 1 ea

Chiral GC Columns
Group 2: Surface/Inclusion Interactions, Simple Derivatives: Permethyl (PM, 110, 120) Cyclodextrin Derivatives
γ-DEX™ 120 Use γ-DEX to reverse elution order for many compounds
(methyl mandelate shown)
Because the elution order of the members of a chiral pair frequently reverses
(enantioreversal) on a γ-DEX column compared to the elution order on an α- R
R
DEX or β-DEX column, we recommend γ-DEX 120 columns as complements
to α-DEX 120 and β-DEX 120 columns. γ-DEX is useful for enantiomeric
differentiation of large analytes, i.e. α-BHC, carvone, carboxylic acids and
α-DEX 120
methamphetamine.
Temp. Limits:
• 30 °C to 230 °C g-DEX 120
phase .................... non-bonded; 20% permethylated γ-cyclodextrin in SPB-35 poly(35% phenyl/65%
dimethylsiloxane)
I.D. Length Beta
0.25 0.25 30 250 24307 1 ea S
S
Resolve positional isomers
p-Xylene (>99%)
20
Min
22 20 22
column: α-DEX 120 and γ-DEX 120, 30 m 3 0.25 mm I.D., 0.25 μm

α-DEX 120 (24310), γ-DEX 120 (24307)
oven: 130 °C
inj.: 250 °C
det.: FID, 300 °C
carrier gas: helium, 35 cm/sec
sample: 1 μL methylene chloride (1 mg/mL each analyte), split (100:1)
m-Xylene
o-Xylene Group 3: Inclusion Interactions
The third group relies on inclusion interactions for retention mechanism.
There are two derivatives in this group:
• Astec CHIRALDEX® DA (Dialkyl derivatives)
• Astec CHIRALDEX® PH (S-Hydroxypropyl derivatives)
The fact that there are three different size cyclodextrins (α, β, and γ) allows
14 16 18
for separation of a wide variety of different size analytes. Astec CHIRALDEX®
Min
B-DA demonstrates the strongest size selectivity. This phase requires
column: α-DEX 120, 30 m 3 0.25 mm I.D., 0.25 μm (24310)
analytes to minimally contain two ring structures, one of which is
oven: 50 °C unsaturated (aromatic). The mechanism of this phase is strongly dependent
inj.: 80 °C on the inclusion mechanism and is able to differentiate changes in the base
det.: FID, 300 °C structure. Because the CHIRALDEX® DA phases most effectively separate
carrier gas: helium, 30 cm/sec multi-ring analytes, analysis temperatures are often higher than 150 °C. A key
sample: 0.6 μL each analyte (neat), split (100:1) application area for this phase is fingerprinting raw materials and identifying
structural differences.
Astec CHIRALDEX® B-PH shows at least some selectivity to a great variety of
analytes, but is especially effective for saturated analytes with minimal
functionality, saturated cyclics, and saturated bicyclics. This phase often
shows a reversal of elution order (enantioreversal) compared to the
CHIRALDEX® B-DA phase.

Chiral GC Columns
Group 3: Inclusion Interactions: Dialkyl (DA) Cyclodextrin Derivatives
Dialkyl (DA) Cyclodextrin Derivatives Astec CHIRALDEX® B-DA Capillary GC Column
Astec CHIRALDEX® A-DA, B-DA, and G-DA CHIRALDEX B-DA requires that analytes possess a minimum of two ring
structures, one of which is unsaturated (aromatic) α, β to the stereogenic
The dipentylated cyclodextrin derivatives show pronounced selectivity center. Examples include fluoxetine, methylphenidate and chlorophenir-
differences based on the size, shape and functionality of the analyte. Strong amine. Inclusion complexation or proper fit between the analyte and
evidence exists for inclusion complexation as the basic driving mechanism cyclodextrin cavity is the dominant enantioselectivity mechanism for the DA
and, therefore, resolution is affected by sample load. series. There must be an includable group α or β to the stereogenic center
The most popular member of this group is Astec CHIRALDEX B-DA. It for chiral recognition. Since CHIRALDEX DA columns most effectively
requires minimally two ring structures, one of which is unsaturated separate multi-ring analytes, analysis temperatures are often higher than
(aromatic) α, β to the stereogenic center (examples include fluoxetine, 150°C. Enantioselectivity has been observed at temperatures >200°C
methylphenidate, chlorpheniramine). Inclusion complexation or proper fit (fluoxetine acetyl derivative).
between the analyte and cyclodextrin cavity is the dominant enantiose- Temp. Limits:
lectivity mechanism for the CHIRALDEX DA series of columns. There must be • -10 °C to 200 °C isothermal, 220 °C programmed
an includable group α or β to the stereogenic center for chiral recognition.
phase ................................................... non-bonded; 2,6-di-O-pentyl-3-methoxy derivative of β-cyclodextrin
Since the Astec CHIRALDEX DA series of columns most effectively separate
multi-ring analytes, analysis temperatures are often higher than 150°C. I.D. Length Beta
Enantioselectivity has been observed at temperatures >200°C (fluoxetine
acetyl derivative). 0.25 0.12 30 500 72023AST 1 ea
Features Astec CHIRALDEX® G-DA Capillary GC Column

• Phase: 2,6-di-O-pentyl-3-methoxy derivative of α-, β-, or γ-cyclodextrin
Incorporates a phase consisting of a 2,6-di-O-pentyl-3-methoxy derivative of
• Hydrophobic surface
γ-cyclodextrin. This phase is good for separations of heterocyclic amines. It
• Pronounced selectivity differences based on analyte size, shape and
has different selectivity from other phases and often shows reversal in
functionality
elution from the PH phases. MAOT = 200 °C isothermal, 220 °C programmed.
• Different selectivity from other cyclodextrin derivatives
• Thermal limit 200/220 °C (isothermal/programmed) GC capillary column
fused silica
Analytes Temp. Limits:
• Useful for separating heterocyclic amines • -10 °C to 200 °C isothermal, 220 °C programmed
Mechanism Observations phase .................................................... non-bonded; 2,6-di-O-pentyl-3-methoxy derivative of γ-cyclodextrin
• Stronger inclusion for CHIRALDEX DA derivatives, therefore, size selectivity I.D. Length Beta
is important. (mm) df (μm) (m) Value Cat. No. Qty
• Critical temperature dependence for enantioselectivity. Above this 0.25 0.12 30 500 72033AST 1 ea
temperature no separation occurs.
Size Selectivity S-Hydroxypropyl (PH) Cyclodextrin Derivatives
• Unlike LC, the size selectivity and chiral recognition applies to both Astec CHIRALDEX® B-PH
aromatic and nonaromatic enantiomers on this phase.
The first general purpose derivative involved substitution of the cyclodextrin
8P R O D U C T S hydroxyl groups with pure "S" hydroxypropyl followed by permethylation.
The surface is hydrophilic in character and the influence of size and shape
selectivity is greatly reduced but not absent. The β-cyclodextrin form has the
Astec CHIRALDEX® A-DA Capillary GC Column broadest applicability.
Incorporates a phase consisting of a 2,6-di-O-pentyl-3-methoxy derivative of Features
α-cyclodextrin. This phase is good for separations of heterocyclic amines. It • Phase: (S)-2-hydroxy propyl methyl ether derivative of β-cyclodextrin
has different selectivity from other phases and often shows reversal in • Good general purpose chiral column
elution from the PH phases. MAOT = 200 °C isothermal, 220 °C programmed. • Separates a wide variety of enantiomers
GC capillary column • Hydrophilic surface
fused silica • Resolves aliphatic, olefinic and aromatic enantiomers
Temp. Limits: • Thermal limit 200/220 °C (isothermal/programmed)
• -10 °C to 200 °C isothermal, 220 °C programmed Analytes
phase ................................................... non-bonded; 2,6-di-O-pentyl-3-methoxy derivative of α-cyclodextrin • Saturated compounds with minimal functionality
I.D. Length Beta • Saturated cyclics and bicyclics
0.25 0.12 30 500 72003AST 1 ea
• Reduced influence of inclusion complexing
• Less size selectivity compared to DA derivatives
• Strong influence of temperature on selectivity
Size Selectivity
• Minimal size selectivity

Chiral GC Columns
Group 3: Inclusion Interactions: S-Hydroxypropyl (PH) Cyclodextrin Derivatives
8P R O D U C T S Guard Columns for Chiral GC Columns

Tubing Treatment Application Max. Temp.
Astec CHIRALDEX® B-PH Capillary GC Column Non-polar (methyl) Low polarity solvents 360 °C
(e.g., alkanes, carbon disulfide, ethers)
CHIRALDEX B-PH shows at least some selectivity to a great variety of Intermediate polarity Intermediate polarity solvents 360 °C
analytes, but is especially effective for saturated analytes with minimal (phenyl/methyl) (e.g., acetone, methylene chloride, toluene)
functionality, saturated cyclics and bicyclics. The CHIRALDEX PH series of
columns shows less of a necessity for inclusion complexation for chiral
Non-Polar Fused Silica Tubing
recognition than the DA columns. This phase often shows a reversal of
elution order (enantioreversal) compared to the B-DA phase. max. temp. ........................................................................................................................................................................................ 360 °C
Temp. Limits: I.D. (mm) L (m) Cat. No. Qty

• -10 °C to 200 °C isothermal, 220 °C programmed 0.25 1 24025 3 ea
0.25 3 25722 1 ea
phase ................................ non-bonded; (S)-2-hydroxy propyl methyl ether derivative of β-cyclodextrin
0.25 5 25742 1 ea
I.D. Length Beta
0.25 15 25756 1 ea
0.25 0.12 30 500 71023AST 1 ea
Intermediate Polar Fused Silica Tubing
max. temp. ........................................................................................................................................................................................ 360 °C
I.D. (mm) L (m) Cat. No. Qty
Chiral GC Column Protection 0.25 3 25727 1 ea
Guard columns protect your capillary column investment. They remove 0.25 5 25747 1 ea
residual moisture, protect the column from non-volatile impurities and the 0.25 15 25760-U 1 ea
high temperature of the injector and/or detector, and allow for the injection
of sample volumes up to 7 μL on-column. Typically a 5-10 m long guard
column is used. The guard column can be connected via a press fit or other
column connector. To couple to a mass spectrometer, use a 1 m length as a
transfer line. We recommend using methyl- or phenyl/methyl-deactivated
guard columns to protect CHIRALDEX® and Supelco DEX™ capillary GC
columns.
Chiral GC Column Test Mixes

After you install a column in your system, use a test mix to make sure you haven’t also installed some surprises (such as ferrule or tubing fragments in the
column, or small leaks). Weekly tests thereafter will keep little problems from growing into big problems. Test mixes are an inexpensive aid to obtaining high
quality chromatograms.
Description Concentration Application Cat. No. Qty

β-DEX™ 120 Column Test Mix 500 μg/mL each component in methylene For use with Supelco β-DEX 120. 48028 1 mL
chloride
Decane (+)-3-Methyl-2-heptanone
3,3-Dimethyl-2-butanol Nonane
1-Hexanol Undecane
α-DEX™ 120 Column Test Mix 500 μg/mL each component in methylene For use with Supelco α-DEX 120. 48013 1 mL
chloride
Decane Undecane
Nonane m-Xylene
1,2-Propanediol p-Xylene
1-(N-TFA)-2-Methylpiperidine 5000 μg/mL in ethanol: isopropanol (95:5) For use with CHIRALDEX G-TA. 90002AST 1 mL
2-(N-TFA)aminoheptane 5000 μg/mL in ethanol: isopropanol (95:5) For use with CHIRALDEX B-PH, A-TA, G-DM 90003AST 1 mL
and G-DP.
1-(N-TFA)aminoindan 5000 μg/mL in ethanol: isopropanol (95:5) For use with CHIRALDEX B-DA and G-DA. 90004AST 1 mL
2-(Bromomethyl)tetra-2H-pyran 5000 μg/mL in ethanol: isopropanol (95:5) For use with CHIRALDEX B-TA and B-DP. 90005AST 1 mL
3,4-Dihydro-2-ethoxy-2H-pyran 5000 μg/mL in ethanol: isopropanol (95:5) For use with CHIRALDEX A-PH and G-PH. 90006AST 1 mL
1-Phenyl-1-ethanol 5000 μg/mL in ethanol: isopropanol (95:5) For use with CHIRALDEX B-PM and B-DM. 90007AST 1 mL

Chiral Derivatization Reagents

ChiraSelect™ is a unique, high-quality set of the most useful chiral derivatization reagents, carefully produced and rigorously analyzed in our laboratories for all
your analytical applications in the chiral field. ChiraSelect reagents are specially selected to meet the requirements for derivatization reagents for enantiomeric
excess determinations. To meet any analytical situation, the ChiraSelect line provides pairs of reagents with each respective enantiomer exhibiting an
enantiomeric ratio of 99.5:0.5.
ChiraSelect™ HPLC Derivatization Reagents

CAS No. Compound Cat. No. Qty
7322-88-5 (S)-(+)-O-Acetylmandelic acid, 99% 253022-5G 5g
- (S)-5-Allyl-2-oxabicyclo[3.3.0]oct-8-ene, purum p.a., chiral derivatization reagent for HPLC, ≥97.0% (GC) 53835-1G 1g
53835-5G 5g
20887-95-0 Boc-Cys-OH, for chiral derivatization, ≥98.5% 15411-250MG 250 mg
15411-1G 1g
19132-06-0 L-(+)-2,3-Butanediol, for chiral derivatization, ≥97.0% 18967-1ML 1 mL
18967-5ML 5 mL
89104-48-3 (4R,5R)-2-Chloro-4,5-dimethyl-1,3,2-dioxaphospholane 2-oxide, for chiral derivatization, ≥95.0% 24370-500MG-F 500 mg
24370-5G-F 5g
28166-41-8 α-Cyano-4-hydroxycinnamic acid, 99% 476870-2G 2g
476870-10G 10 g
130678-42-1 (+)-Diisopropyl-O,O′-bis(trimethylsilyl)-L-tartrate, 99%, Flukabrand™ ChiraSelect reagent 420131-1G 1g
95713-52-3 Nα-(2,4-Dinitro-5-fluorophenyl)-L-alaninamide, for chiral derivatization, ≥99.0% 71478-50MG 50 mg
210529-62-7 Nα-(2,4-Dinitro-5-fluorophenyl)-D-valinamide, for chiral derivatization, ≥98.0% 42100-500MG 500 mg
132679-61-9 Nα-(2,4-Dinitro-5-fluorophenyl)-L-valinamide, for chiral derivatization, ≥98.0% 42102-100MG 100 mg
69632-32-2 (R)-(−)-3,5-Dinitro-N-(1-phenylethyl)benzamide, 98% 296902-1G 1g
69632-31-1 (S)-(+)-3,5-Dinitro-N-(1-phenylethyl)benzamide, 98% 296910-1G 1g
154479-90-0 (−)-1-(9-Fluorenyl)ethyl chloroformate solution, 18 mM in acetone, for chiral derivatization 338710-1ML 1 mL
107474-79-3 (+)-1-(9-Fluorenyl)ethyl chloroformate solution, ≥18 mM in acetone, for chiral derivatization 23182-10X1ML-F 10 × 1 mL
23182-10ML-F 10 mL
124529-07-3 N-Isobutyryl-D-cysteine, for chiral derivatization, ≥97.0% 58689-250MG 250 mg
124529-02-8 N-Isobutyryl-L-cysteine, for chiral derivatization, ≥97.0% 58698-250MG-F 250 mg
58698-1G-F 1g
784213-51-0 (1R,4aS,10aR)-7-Isopropyl-1-isothiocyanato-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene, for chiral 89394-100MG 100 mg
derivatization
3966-32-3 (R)-(−)-α-Methoxyphenylacetic acid, for chiral derivatization, ≥99.0% 65209-1G 1g
26164-26-1 (S)-(+)-α-Methoxyphenylacetic acid, for chiral derivatization, ≥99.0% 65208-250MG 250 mg
139658-04-1 (R)-6-Methoxy-2,5,7,8-tetramethylchromane-2-carboxylic acid, for chiral derivatization, ≥99.0% 93509-50MG 50 mg
24277-44-9 (−)-α-Methylbenzyl isothiocyanate, for chiral derivatization, ≥99.0% 89568-250MG-F 250 mg
24277-43-8 (S)-(+)-α-Methylbenzyl isothiocyanate, for chiral derivatization, ≥99.0% 75491-1G-F 1g
10420-89-0 (S)-(−)-1-(1-Naphthyl)ethylamine, ≥99% 237450-1G 1g
237450-5G 5g
73671-79-1 (S)-(+)-1-(1-Naphthyl)ethyl isocyanate, 99% 295957-250MG 250 mg
295957-1G 1g
159717-68-7 N-(7-Nitro-4-benzofurazanyl)-L-prolyl chloride, for fluorescence 84999-50MG-F 50 mg
5978-70-1 (R)-(−)-2-Octanol, for chiral derivatization, 99% 147990-1G 1g
147990-5G 5g
147990-10G 10 g
6169-06-8 (S)-(+)-2-Octanol, 99% 147982-5G 5g
147982-10G 10 g
7782-24-3 (S)-(+)-2-Phenylpropionic acid, 97% 279900-250MG 250 mg
279900-1G 1g
- Quaternary β-cyclodextrin, 100mg, neat 33805 100 mg
- Sulphated β-cyclodextrin, 100mg, neat 33806 1 amp
14152-97-7 2,3,4,6-Tetra-O-acetyl-β-D-glucopyranosyl isothiocyanate, for HPLC derivatization T5783-100MG 100 mg
T5783-1G 1g
- 2,3,4,6-Tetra-O-(2-naphthoyl)-β-D-galactopyranosyl isothiocyanate, for derivatization, ~90% (HPLC) 04669-25MG-F 25 mg
04669-100MG-F 100 mg
147948-52-5 2,3,4,6-Tetra-O-pivaloyl-β-D-galactopyranosyl isothiocyanate, for chiral derivatization, ≥95% (HPLC, sum of 88102-100MG 100 mg
enantiomers) 88102-500MG 500 mg
958300-06-6 2,3,4,6-Tetra-O-pivaloyl-β-D-glucopyranosyl isothiocyanate, for chiral derivatization, ≥95.0% (HPLC) 44891-100MG-F 100 mg
62414-75-9 2,3,4-Tri-O-acetyl-α-D-arabinopyranosyl isothiocyanate, for chiral derivatization, ≥98.0% 90245-100MG 100 mg
10531-50-7 (R)-(−)-α-(Trifluoromethyl)benzyl alcohol, puriss., ≥99.0% (GC, sum of enantiomers) 79231-1ML 1 mL
340-06-7 (S)-(+)-α-(Trifluoromethyl)benzyl alcohol, 99% 411140-250MG 250 mg
411140-1G 1g
14645-24-0 (−)-Tröger’s base, for chiral derivatization, ≥99.0% 40765-100MG 100 mg
21451-74-1 (+)-Tröger’s base, for chiral derivatization, ≥99.0% 40764-100MG 100 mg
135806-59-6 (S)-Trolox methyl ether, for chiral derivatization, ≥98.0% 93510-50MG 50 mg


ChiraSelect™ GC Derivatization Reagents
ChiraSelect™ GC Derivatization Reagents

36394-75-9 (S)-(−)-2-Acetoxypropionyl chloride, for chiral derivatization, ≥99.0% 00877-500MG 500 mg
104530-16-7 (1R)-(+)-Camphanic chloride, for chiral derivatization, ≥97.0% 21286-250MG-F 250 mg
39637-74-6 (1S)-(−)-Camphanic chloride, for chiral derivatization, ≥98.0% 21287-1G-F 1g
21287-5G-F 5g
21287-25G-F 25 g
3347-90-8 (S)-2-Hydroxybutyric acid, for chiral derivatization, ≥97.0% 54918-1G-F 1g
20445-31-2 (R)-(+)-α-Methoxy-α-trifluoromethylphenylacetic acid, for chiral derivatization, ≥99.0% 65361-250MG 250 mg
65361-1G 1g
17257-71-5 (S)-(−)-α-Methoxy-α-trifluoromethylphenylacetic acid, for chiral derivatization, ≥99.0% 65369-250MG-F 250 mg
39637-99-5 (R)-(−)-α-Methoxy-α-(trifluoromethyl)phenylacetyl chloride, for chiral derivatization, ≥99.0% 65363-100MG 100 mg
65363-500MG 500 mg
20445-33-4 (S)-(+)-α-Methoxy-α-trifluoromethylphenylacetyl chloride, for chiral derivatization, ≥99.0% 65365-100MG-F 100 mg
65365-500MG-F 500 mg
3886-69-9 (R)-(+)-α-Methylbenzylamine, for chiral derivatization, ≥99.0% 77879-5ML 5 mL
77879-25ML 25 mL
2627-86-3 (S)-(−)-α-Methylbenzylamine, for chiral derivatization, ≥99.0% 77869-5ML 5 mL
77869-25ML 25 mL
33375-06-3 (R)-(+)-α-Methylbenzyl isocyanate, for chiral derivatization, ≥99.0% 77968-1ML 1 mL
77968-5ML 5 mL
14649-03-7 (S)-(−)-α-Methylbenzyl isocyanate, for chiral derivatization, ≥99.0% 77970-1ML 1 mL
77970-5ML 5 mL
104371-21-3 (R)-(+)-α-Methyl-2,3,4,5,6-pentafluorobenzyl alcohol, for chiral derivatization, ≥99.0% 76744-1G 1g
104371-20-2 (S)-(−)-α-Methyl-2,3,4,5,6-pentafluorobenzyl alcohol, for chiral derivatization, ≥99.0% 76746-1G 1g
3886-70-2 (R)-(+)-1-(1-Naphthyl)ethylamine, for chiral derivatization, ≥99.5% 70710-1ML 1 mL
42340-98-7 (R)-(−)-1-(1-Naphthyl)ethyl isocyanate, for chiral derivatization, ≥99.0% 70725-1ML 1 mL
1517-69-7 (R)-(+)-1-Phenylethanol, for chiral derivatization, ≥99.0% 77848-1ML 1 mL
77848-5ML 5 mL
1445-91-6 (S)-(−)-1-Phenylethanol, for chiral derivatization, ≥99.0% 77849-1ML 1 mL
77849-5ML 5 mL

Chiral Mobile Phase Additives
Chiral Mobile Phase Additives

Enantiomers of the same parent compound differ in the way they interact with other chiral molecules, like chiral stationary phases. However, it is possible to
affect an enantiomeric separation using conventional HPLC and CE stationary phases by adding the chiral selector to the mobile phase (1,2). These chiral
selector additives generally interact via ion pair, ligand exchange or inclusion interactions with enantiomer analytes, forming mobile diastereomeric complexes
that are therefore separable by conventional normal or reversed-phase columns. When free cyclodextrins are added to the mobile phase, inclusion complexes
are formed and separations can approach those obtained on cyclodextrin-based CSPs. Note that this approach often leads to a reversal in the elution order
obtained on a cyclodextrin CSP. Sigma-Aldrich carries a number of highly-pure cyclodextrin derivatives for this application.
(1) Armstrong, D. W. Pseudophase Liquid Chromatography: Applications to TLC. J. Liq. Chromatogr. 1980, 3(6), 895-900.
(2) Ameyibor, E.; Stewart, J. T. Enantiomeric HPLC Separation of Selected Chiral Drugs Using Native and Derivatized ß-Cyclodextrins as Chiral Mobile Phase
Additives. J. Liq. Chromatogr. 1997, 20(6), 855-869.

- (2-Carboxyethyl)-β-cyclodextrin sodium salt 21872-1G-F 1g
21872-5G-F 5g
- Carboxymethyl-β-cyclodextrin sodium salt 21906-1G 1 g
21906-5G 5 g
10016-20-3 α-Cyclodextrin, purum, ≥98.0% (HPLC) 28705-5G 5 g
28705-25G 25 g
28705-100G 100 g
68168-23-0 β-Cyclodextrin hydrate, 99% 856088-5G 5g
856088-25G 25 g
856088-100G 100 g
68168-23-0 β-Cyclodextrin hydrate, puriss., ≥99.0% (HPLC) 28707-5G 5g
28707-25G 25 g
28707-100G 100 g
91464-90-3 γ-Cyclodextrin hydrate 861413-100MG 100 mg
861413-1G 1g
699020-02-5 α-Cyclodextrin, sulfated sodium salt hydrate 494542-5G 5g
37191-69-8 β-Cyclodextrin, sulfated sodium salt, extent of labeling: 7-11 mol per mol β-CD 389153-5G 5g
389153-25G 25 g
51166-71-3 Heptakis(2,6-di-O-methyl)-β-cyclodextrin H0513-1G 1g
H0513-5G 5g
51166-71-3 Heptakis(2,6-di-O-methyl)-β-cyclodextrin, ≥98.0% (TLC) 39915-1G 1g
55216-11-0 Heptakis(2,3,6-tri-O-methyl)-β-cyclodextrin, ≥90% H4645-5G 5g
55216-11-0 Heptakis(2,3,6-tri-O-methyl)-β-cyclodextrin, ≥98.0% 51707-1G 1g
51707-5G 5g
128446-32-2 (2-Hydroxyethyl)-β-cyclodextrin, extent of labeling: ~0.7 mol per mol cellulose 389137-10G 10 g
128446-33-3 (2-Hydroxypropyl)-α-cyclodextrin, average Mw ~1,180 390690-5G 5 g
390690-25G 25 g
128446-35-5 (2-Hydroxypropyl)-β-cyclodextrin, average Mw ~1,380 332593-5G 5 g
332593-25G 25 g
332593-100G 100 g
128446-35-5 (2-Hydroxypropyl)-β-cyclodextrin, average Mw ~1,460 332607-5G 5 g
332607-25G 25 g
332607-100G 100 g
332607-500G 500 g
128446-35-5 (2-Hydroxypropyl)-β-cyclodextrin, average Mw ~1,540 389145-5G 5g
389145-25G 25 g
128446-35-5 (2-Hydroxypropyl)-β-cyclodextrin, Mr ~1380 56332
128446-34-4 (2-Hydroxypropyl)-γ-cyclodextrin, solid H125-5G-I 5g
H125-100G-I 100 g
128446-34-4 (2-Hydroxypropyl)-γ-cyclodextrin, extent of labeling: 0.6 molar substitution 390704-5G 5g
390704-25G 25 g
128446-36-6 Methyl-β-cyclodextrin, average Mn 1310 332615-5G 5g
332615-25G 25 g
- Succinyl-β-cyclodextrin 85990-500MG 500 mg
85990-5G 5g
23739-88-0 Triacetyl-β-cyclodextrin 332623-10G 10 g
AUTHORIZED DISTRIBUTOR

Supelco 3 Chiral PDF

Uploaded by

Copyright:

Available Formats

Supelco 3 Chiral PDF

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Supelco 3 Chiral PDF

Uploaded by

Copyright:

Available Formats

Table of Contents 5

For all of your analytical needs, visit us at sigma-aldrich.com/analytical

Think Chiral...Think Supelco

sigma-aldrich.com/analytical View our current seminar and tradeshow schedule at sigma-aldrich.com/analytical-events

Method Development and Column Screening Protocol on CHIRO-

• Astec CHIROBIOTIC® macrocyclic glycopeptide-based CSPs

For hazardous product information, visit sigma-aldrich.com/safetycenter

Chiral HPLC & SFC Columns

• A number of the Astec CYCLOBOND derivatives offer good opportunities

CHIROBIOTIC® CSPs—Physical Properties

Proposed structure of vancomycin (the chiral selector in CHIROBIOTIC® V and V2)

sigma-aldrich.com/analytical Access our analytical literature and newsletters at sigma-aldrich.com/literature

Chiral HPLC & SFC Columns

 application for HPLC

t-Bu H3C N HBr

t-Bu t-Bu CH3

Application courtesy of Dr. M. Althous,

Demonstration of CHIROBIOTIC® column utility in both normal phase

Astec CHIROBIOTIC® Application Areas

Simplified Chiral Method Development

View our technical presentations at sigma-aldrich.com/analytical-videos

Chiral HPLC & SFC Columns

Complementary Selectivity to Cellulosic/Amylosic HPLC Analysis of Clenbuterol Enantiomers on Astec® CHIRO-

24 because separations on Astec CHIROBIOTIC® CSPs are usually very fast.

sigma-aldrich.com/analytical To order, visit sigma-aldrich.com/order

Chiral HPLC & SFC Columns

For hazardous product information, visit sigma-aldrich.com/safetycenter

Chiral HPLC & SFC Columns

Astec CHIROBIOTIC® T Chiral HPLC Column Astec CHIROBIOTIC® R (Ristocetin A)

Astec CHIROBIOTIC® T2 Chiral HPLC Column

sigma-aldrich.com/analytical For all of your analytical needs, visit us at sigma-aldrich.com/analytical

Chiral HPLC & SFC Columns

Astec CHIROBIOTIC® TAG Chiral HPLC Column

I.D. (mm) L (cm) Cat. No. Qty No. Title

Representation of a cyclodextrin toroid molecule attached to a silica surface. The functionalized

For technical assistance, visit sigma-aldrich.com/techinfo

Chiral HPLC & SFC Columns

CYCLOBOND is the name given to the Astec technology for bonding

sigma-aldrich.com/analytical View our current seminar and tradeshow schedule at sigma-aldrich.com/analytical-events

Chiral HPLC & SFC Columns

• Bonded phase: Dimethylated β-cyclodextrin 4.6 25 22724AST 1 ea

For hazardous product information, visit sigma-aldrich.com/safetycenter

Chiral HPLC & SFC Columns

sigma-aldrich.com/analytical Access our analytical literature and newsletters at sigma-aldrich.com/literature

Chiral HPLC & SFC Columns

View our technical presentations at sigma-aldrich.com/analytical-videos

Chiral HPLC & SFC Columns

Columns For other column dimensions not listed, please inquire.

sigma-aldrich.com/analytical To order, visit sigma-aldrich.com/order

Chiral HPLC & SFC Columns

Astec CLC-L Chiral HPLC Column

We recommend using a Supelco ColumnSaver precolumn filter (Cat. No.

For hazardous product information, visit sigma-aldrich.com/safetycenter

Chiral HPLC & SFC Columns

CHIRALPAK® AGP HPLC Column

sigma-aldrich.com/analytical For all of your analytical needs, visit us at sigma-aldrich.com/analytical

Chiral HPLC & SFC Columns

For technical assistance, visit sigma-aldrich.com/techinfo

Chiral HPLC & SFC Columns