5µm 3

µm

5 Frequently Asked Question About Lux®

Chiral Columns

phenomenex.com/Lux

GR

6796

0419

How can I reverse enantiomeric elution order?

Frequently Asked Chiral Questions

Q1:

App

ID 2

5337

OO

H3C

Cl

NH

Amylose

Batch A

0 5 10 15 20 min

mAU

0

100

200

300

400

NEW Lux i-Amylose-3

App

ID 2

5366

Cellulose

OO

CI

CI

NH

0 5 10 15 20 min

mAU

0

200

400

600

800

OO

H3C

CH3

NH

Amylose

App

ID 2

5367

0 5 10 15 20 min

mAU

0

100

200

300

400

Lux i-Amylose-1

Enantiomers of Diniconazole

Cl

Cl

H

OH

N N

N

CH3

CH3

CH3

E

S

Cl

Cl

H

OH

N N

N

CH3

CH3

CH3

E

R

Columns: Lux 5 µm i-Amylose-3Lux 5 µm i-Cellulose-5Lux 5 µm i-Amylose-1

Dimensions: 250 x 4.6 mm Part No.: 00G-4779-E0

00G-4756-E000G-4762-E0

Mobile Phase: Water with 0.1 % Diethylamine/Acetonitrile (35:65)Flow Rate: 1.0 mL/min

Injection Volume: 10 µL (2 mg/mL) Detection: UV @ 254 nm

Sample: 1. Diniconazole 2. Diniconazole

Because of the intrinsic nature of enantiomers, it is difficult to predict selectivity and therefore the relative analyte elution order. However, the Lux® polysaccharide chiral column portfolio contains a wide range of chiral stationary phases that can easily be screened for complementary or orthogonal selectivity. For instance, Lux i-Amylose-3 and i-Cellulose-5 have complementary but distinct selectivity in comparison to each other. In addition, the phase immo-bilization affords greater solvent flexibility, increasing the potential for enantioselectivity.

Lux i-Cellulose-5

Why does Phenomenex use DEA over TEA as the primary basic modifier in application notes?

Frequently Asked Chiral Questions

Q2:

Need a chiral method right now?Start your chiral method search by an analyte

name or structure search in seconds here:

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Both DEA (diethylamine) and TEA (triethylamine) are widely published as good basic modifiers for improving peak shapes on polysaccharide-type chiral columns. We chose DEA for our initial screening data and have continued with it routinely to maintain consistency. TEA is also just as effective and commonly used by many customers successfully on our Lux® polysaccharide chiral columns.

min0 2 4 6 8 10 12 14 16 18

mAU

123456789

min2 4 6 8 10 12 14 16 18

min2 4 6 8 10 12 14 16 18

App

ID 2

5342

Columns: Lux 5 µm i-Amylose-3Dimensions: 250 x 4.6 mm

Part No.: 00G-4779-E0Mobile Phase: Acetonitrile with 0.1 % Diethylamine

Flow Rate: 1.0 mL/min Injection Volume: 10 µL (2 mg/mL)

Detection: UV @ 254 nm Sample: 1. Thalidomide

2. Thalidomide

Enantiomers of Thalidomide

N

NH

O

OCH2

CH2

S N

NH

O

OCH2

CH2

R

Batch B

Batch C

Batch D

min0 2 4 6 8 10 12 14 16 18

mAU

2

4

6

8

10

12

0

min2 4 6 8 10 12 14 16 18

min2 4 6 8 10 12 14 16 18

App

ID 2

5340

Enantiomers of Econazole

Cl

O

Cl

Cl

H

N

N

S

Cl

O

Cl

Cl

H

N

N

R

Batch A

Batch B

Batch C

Columns: Lux® 5 µm i-Amylose-3Dimensions: 250 x 4.6 mm

Part No.: 00G-4779-E0Mobile Phase: Water with 5 mM Ammonium Acetate + 0.05 %

Formic Acid/Acetonitrile (65:35) Flow Rate: 1.0 mL/min

Injection Volume: 10 µL (2 mg/mL) Detection: UV @ 254 nm

Sample: 1. Econazole 2. Econazole

How does the aromaticity of chiral compounds affect selectivity?

Frequently Asked Chiral Questions

Q3: In chiral compounds, the proximity of aromatic groups to the stereocenter is typically linked to the ease of enan-tiomeric resolution. For instance, the separations of enantiomers in which the aromatic functionality is 4 or more atoms away from the stereocenter can be challenging and chromatographically uncommon. Enantioselectivity is most effective when the distances between the aromatic group and stereocenter are equivalent in both chiral conformations. If the aromatic group of the compound has electron withdrawing groups like halogens or oxygen it will be more electron deficient and will interact more effectively with electron-rich aromatic groups of the chiral stationary phase.

What is the difference between Amylose and Cellulose polysaccharide backbones?Q4:

Both backbones form 3-dimensional helical structures that are well suited to provide grooves and cavities for po-tential steric interactions, Hydrogen bonding, dipole-dipole, and π-π based interactions. However, the structure for amylose is considered more tightly coiled in comparison to the looser cellulose structure which may accommodate interactions differently. Practically, this difference can result in distinct selectivity for amylose and cellulose even if with an identical chiral selector, as the 3-dimensional orientation around the CSP will differ.

App

ID 2

5368

3

min0 5 10 15 20 25 min 35 40 45

mAU

0

10

20

30

40

50

60

App

ID 2

5369

5

min0 5 10 15 20 25 min 35 40 45

mAU

01020304050607080

mAU

App

ID 2

5370

min0 5 10 15 20 min 35 40 45

mAU

0

20

40

60

80

100

Enantiomers of Napropamide

O

CH3N

O

CH3

CH3

S

O

CH3N

O

CH3

CH3

R

Columns: Lux® 5 µm i-Amylose-3Lux 5 µm i-Cellulose-5 Lux 5 µm i-Amylose-1

Dimensions: 250 x 4.6 mm Part No.: 00G-4779-E0

00G-4756-E000G-4762-E0

Mobile Phase: Water with 5 mM Ammonium Acetate + 0.05% Formic Acid/Acetonitrile (53:47)

Flow Rate: 1.0 mL/min Injection Volume: 10 µL (2 mg/mL)

Detection: UV @ 254 nm Sample: 1. Napropamide

2. Napropamide

Frequently Asked Chiral Questions

What separation modes can Lux® polysaccharide chiral col-umns operate in?Q5:

Frequently Asked Chiral Questions

Lux chiral columns can be used in reversed phase, normal phase, polar organic, polar ionic and SFC conditions. However, the immobilization and bonding technology used within the Lux i-Amylose-3, i-Cellulose-5 and i-Amy-lose-1 columns promotes stability in strong organic solvents, which affords you the ability to expand your chiral separation success with even more solvent systems.

5µm

Column: Lux 5 µm i-Amylose-3Dimensions: 250 x 4.6 mm

Part No.: 00G-4779-E0Mobile Phase: Hexane/Isopropanol with 0.1 %

Diethylamine (80/20)Flow Rate: 0.5 mL/min

Injection Volume: 10 µL (2 mg/mL) Detection: UV @ 220 nm

Sample: 1. Trans-Stilbene Oxide 2. Trans-Stilbene Oxide

*Aggressive solvent stability was tested by flushing columns with DCM followed by THF before rerunning in the mobile phase.

Before Exposure to Strong Solvents (DCM & THF)*After Exposure

App

ID 2

5392

0 2 4 6 8 10 12 14

0

20

40

60

80

100

mAU

min

Terms and Conditions - Subject to Phenomenex Standard Terms & Conditions, which may be viewed at www.phenomenex.com/TermsAndConditions • Trademarks - Lux is a registered trademark, SecurityLINK, and SecurityGuard are trademarks of Phenomenex. • Disclaimer - Comparative separations may not be representative of all applications. SecurityGuard is patented by Phenomenex. U.S. Patent No. 6,162,362. • CAUTION: this patent only applies to the analytical-sized guard cartridge holder, and does not apply to SemiPrep, PREP or ULTRA holders, or to any cartridges. • FOR RESEARCH USE ONLY. Not for use in clinical diagnostic procedures. • © 2020 Phenomenex, Inc. All rights reserved.