HPLC Column for Structual Isomers - obrnutafaza.hr · Column Size Mobile Phase Flow Rate Temperature Detection 4.6 mm I.D. x 150 mm PYE: Methanol : Water = 90 : 10 NPE: Methanol :

HPLC Column for Structual Isomers

Due to the planar pyrene ring structure and strong - interactions, COSMOSIL PYE achieves excellent separation of aromatic isomers.

Column SizeMobile PhaseFlow RateTemperatureDetection

4.6 mm I.D. x 150 mmMethanol : Water = 90 : 101.0 ml/min30UV 254 nm

Sample 1. Benz [a] Anthracene2. Chrysene

Column SizeMobile Phase

Flow RateTemperatureDetection

Sample

4.6 mm I.D. x 150 mmA: Water, B: MethanolB 80% B 100%20 min. Linear Gradient1.0 ml/min30UV 254 nm

1. Naphthalene2. Acenaphthylene3. Acenaphthene4. Flourene5. Phenathrene6. Acnthracene7. Fluoranthene8. Pyrene9. 2, 3-Benzofluorene10. Benz [a] Anthracene11. Chrysene12. Triphenylene13. Naphthacene14. Benzo[a] Pyrene15. Perylene16. Dibenz [a,c] Anthracene17. Dibenz [a,h] Anthracene

1. Benz [a] anthracene 2. Chrysene

Separation of PAHs

Conventional ODS

COSMOSIL 5PYE

COSMOSIL PYE (Pyrenylethyl group bonded) and COSMOSIL NPE (Nitrophenylethyl group bonded) column show unique retention characteristics based on multiple separation modes such as hydrophobic, charge transfer and - interactions. These columns are recommended for the separation of structural isomers.

COSMOSIL PYE column is a reversed phase column with 2-(1-Pyrenyl) ethyl groups bonded silica packing material. This column utilizes - interactions originating from the planar pyrene ring structure to separate structural isomers.

High Purity Porous Spherical Silica5 m

approx. 120

Silica GelAverage Particle SizeAverage Pore SizeStationary PhaseMain Interaction

Carbon Content

COSMOSIL PYE

2-(1-Pyrenyl)ethyl GroupHydrophobic Interaction

- InteractionCharge Transfer Interaction

Stereoselectivityapprox. 18%

COSMOSIL NPE

Nitrophenylethyl GroupHydrophobic Interaction

- InteractionDipole-Dipole Interaction

approx. 9%

SiH3C CH3 SiH3C CH3

NO2

PYE NPE

Pyrenylethyl group bonded stationary phase Separation with high molecular shape selectivity or - interactions Excellent separation for structural isomers

Application Data Separation of Sterols

Separation of Phthalonitriles

Separation of Tocopherols



4.6 mm I.D. x 150 mm5C18-MS-II NAP

5PYE 1.0 ml/min30UV 210 nm

Methanol : H2O = 98 : 2 Methanol : H2O = 90 : 10 Methanol : H2O = 95 : 5

Sample (5.6 g)(5.9 g)

1. Cholesterol 2. Stigmasterol 3. Campesterol4. Sitosterol

5C18-MS-II NAP 5PYE

3

CH3

3

H3C CH3

HO

CH3

CH3R

H3C CH3

CH3

CH3

H3C CH

CH3

2; H3C CHCH3

CH3

3;

4;

CH31;

R=

5C18-MS-II NAP 5PYE



4.6 mm I.D. x 150 mm5C18-MS-II NAP


Methanol : H2O = 30: 70 Methanol : H2O = 50 : 50 Methanol : H2O = 80 : 20

Sample Phthalonitrile IsophthalonitrileTerephthalonitrile

o-; m-; p-;

(0.3 g)(3.0 g)(0.15 g)

CN

CN

CN

CN

CN

CNo- m- p-

5C18-MS-II NAP 5PYE



Sample4.6 mm I.D. x 150 mm5C18-MS-II NAP


Methanol : H2O = 95 : 5 Methanol : H2O = 90 : 10 Methanol : H2O = 90 : 10

-Tocopherol (R1:Me, R2:Me)-Tocopherol (R1:Me, R2:H)-Tocopherol (R1:H, R2:Me)-Tocopherol (R1:H, R2:H)

H

R 2

O

CH3

3C

R1

H3C H

HO

H3C H

CH3

CH3

Separation of Capsaicin

O

HO

CH3

HN

O

H

HCH3

CH3Capsaicin

HN

O

HO

CH3 O

CH3N-Vanillylnonanamide

Column sizeMobile phaseFlow rateTemperatureDetection


(1.0 g)(0.5 g)

Sample1. Capsaicin2. N-Vanillylnonanamide

5PYE5C18-MS-II NAP

o,p-

m- m-o-

p-

o-m-

p-

COSMOSIL PYE (continued)



4.6 mm I.D. x 150 mmPYE: Methanol : Water = 90 : 10NPE: Methanol : Water = 70 : 30ODS: Methanol : Water = 80 : 201.0 ml/min30UV 254 nm

COSMOSIL NPE strongly retains 1,8-dinitronaphthalene because of the strong dipole formed by the two nitro groups positioned on the same side of naphthalene.

Sample1. 1,8-Dinitronaphtalene2. 1,5-Dinitronaphtalene3. Naphtalene4. 1-Methylnaphtalene5. 1,5-Dimethylnaphtalene

NO2 NO2

NO2

NO2 CH3 CH3

CH31 2 3 4 5

Application of Disubstituted Naphtalenes

COSMOSIL NPE column is a reversed phase column with Nitrophenylethyl groups bonded silica packing material. This columnprovides unique retention characteristics, slightly different from the COSMOSIL PYE column, utilizing both dipole-dipole and - interactions.

T

m-

p-

o-

B

The chromatogram below illustrates the separation of o-, m-, p-tolunitrile. Since hydrophobic interaction is dominant in the separation by a ODS column, tolunitrile elutes first. In COSMOSIL NPE, tolunitrile elutes later. This suggests that COSMOSIL NPE utilizes the interaction between -electron of the nitrophenyl group and CN- for retention in addition to hydrophobic interac-tion.

Column SizeMobile PhaseFlow RateTemperatureDetection


CH3

CN

CH3

Tolunitrile(o-, m-, p-)

Benzene(B)

Toluene(T)

T

m-

o-, p-

B

Advantage of Dipole-Dipole Interactions

5NPE

5PYE 5NPE 5C18-MS-II

Nitrophenylethyl group bonded stationary phase Separation with dipole-dipole and - interactions Excellent separation for structural isomers

5C18-MS-II

5NPE 5PYE

The ODS column cannot sharply separate the positional isomers like phthalonitriles. In contrast, COSMOSIL PYE and NPE can separate them very well by - interaction. Furthermore, COSMOSIL NPE strongly retains ortho compound which has a big dipole moment.


Flow RateTemperatureDetectionSample

4.6 mm I.D. x 150 mmNPE: Methanol : Water = 40 : 60PYE: Methanol : Water = 80 : 20ODS: Methanol : Water = 30 : 701.0 ml/min30UV 254 nm1. o-Phthalonitrile2. m-Phthalonitrile3. p-Phthalonitrile

(0.30 g)(3.00 g)(0.15 g)

o- p-m-

CN

CN

CN

CN

CNCN

5C18-MS-II

Effect of Dipole-Dipole Interaction

m-

p-

o-

o-m-

p-

o, p-

m-

NACALAI TESQUE, INC.Nijo Karasuma, Nakagyo-kuKyoto 604-0855, JAPANTELFAXWebsiteE-mail

For research use only, not intended for diagnostic or drug use.

1004

: +81-(0)75-251-1730: +81-(0)75-251-1763: http://www.nacalai.com: [email protected]

Other size may be available. Please enquire.

Application Data

Column SizeMobile PhaseFlow RateTemperatureDetectionSample

4.6 mm I.D. x 150 mmMethanol : 20 mmol/l KH2PO4 = 60 : 401.0 ml/min30UV 254 nm1. 2-Benzylpyridine2. 3-Benzylpyridine3. 4-Benzylpyridine

5PYE

Column SizeMobile PhaseFlow RateTemperatureDetectionSample

4.6 mm I.D. x 150 mm70% Methanol1.0 ml/min30UV 215 nm1. Carvylacetate (cis, trans)

5PYE

OCOCH3

N

Positional Isomers like Benzylpyridines

Geometrical Isomers like Carvylacetates

5NPE

5NPE

Product NameCOSMOSIL 5PYEPacked Column

COSMOSIL 5NPEPacked Column

Column Size1.0 mm I.D. x 150 mm2.0 mm I.D. x 150 mm2.0 mm I.D. x 250 mm4.6 mm I.D. x 150 mm4.6 mm I.D. x 250 mm10.0 mm I.D. x 250 mm20.0 mm I.D. x 250 mm1.0 mm I.D. x 150 mm2.0 mm I.D. x 150 mm2.0 mm I.D. x 250 mm4.6 mm I.D. x 150 mm4.6 mm I.D. x 250 mm10.0 mm I.D. x 250 mm20.0 mm I.D. x 250 mm

Product Number02851-7138042-6134450-3137837-9137989-1137996-1138044-4105897-0134328-5134379-9137902-2137990-7105469-1138046-21

Product NameCOSMOSIL 5PYEGuard Column

COSMOSIL 5NPEGuard Column

Ordering Information

COSMOSIL NPE (continued)

Column Size4.6 mm I.D. x 10 mm10.0 mm I.D. x 20 mm20.0 mm I.D. x 20 mm20.0 mm I.D. x 50 mm4.6 mm I.D. x 10 mm10.0 mm I.D. x 20 mm20.0 mm I.D. x 20 mm20.0 mm I.D. x 50 mm

Product Number37903-1138041-7105867-9134475-2137904-0138045-3105868-8105869-71

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COSMOSIL PYEPyrenylethyl group bonded stationary phase

Excellent separation for structural isomers

Applications

0.0 1.0 2.0 2.5

Nitrobenzene/Benzene), 50%Methanol

3.01.5

Cholester

C18-MS-

PE-MS

NAP

PYE

0.5

-

COSMOSIL Application DataCOSMOSIL Application Data

NACALAI TESQUE, INC

Column: Column size: 4.6mmI.D.-150mmMobile phase: 5C18-MS-II Methanol/ H2O = 98/2

NAP Methanol/ H2O = 90/105PYE Methanol/ H2O = 95/5

Flow rate: 1.0 ml/min Temperature: 30 CDetection: UV210nm

Sample: 1; Cholesterol (5.6 g) 2; Stigmasterol (5.9 g) 3; Campesterol4; Sitosterol

AP-1026

5C18-MS-II 5PYENAP

HO

CH3

CH3R

H3C CH3

CH3

CH3

H3C CH3

CH3

CH3

2; H3C CH3CH3

CH3

3;

4;

H3C CH3

CH31;

R=


NACALAI TESQUE, INC




Sample: o-; Phthalonitrile (0.3 g) m-; Isophthalonitrile (3.0 g) p-; Terephthalonitrile (0.15 g)

AP-1027

5C18-MS-II 5PYENAP

CN

CN

CN

CN

CN

CNo- m- p-

-

PE-MS PYE

SiH3C CH3

SiH3C CH3

SiH3C CH3

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Applications

Attention1. Methanol is recommended as a mobile phase for COSMOSIL PYE column. Acetonitrile is not recommended because it has

-

2. The stationary phase of COSMOSIL PYE, pyrenylethyl group, has a large UV absorption. When the stationary phase detaches

from silica gel and elutes, even a slight quantity can be detected and causes baseline noise. In such a case, wash the column

with tetrahydrofuran. Detachment of a small amount of the stationary phase does not deteriorate a columns separation ability.

3. COSMOSIL PYE column is not suitable for gradient analysis.

Ordering Information


NACALAI TESQUE, INC

Column: Column size: 4.6mmI.D.-150mmMobile phase: Methanol/ H2O = 70/30 Flow rate: 1.0 ml/min Temperature: 30 CDetection: UV280nm

Sample: 1; Capsaicin (1.0 g) 2; N-Vanillylnonanamide (0.5 g)

AP-1073

5C18-MS-II 5PYENAP

O

HO

CH3

HN

O

H

HCH3

CH3

HN

O

HO

CH3 O

CH3

Capsaicin

N-Vanillylnonanamide

COSMOSIL 5PYE Packed ColumnColumn Size

I.D. x Length (mm)1.0 x 150 02851-712.0 x 150 38042-612.0 x 250 34450-31

Column SizeI.D. x Length (mm)

4.6 x 150 37837-914.6 x 250 37989-1110 x 250 37996-1120 x 250 38044-41

COSMOSIL 5PYE Guard ColumnColumn Size

I.D. x Length (mm)4.6 x 10 37903-1110 x 20 38041-7120 x 20 05867-9120 x 50 34475-21


NACALAI TESQUE, INC




Sample: -Tocopherol-Tocopherol-Tocopherol-Tocopherol

AP-1028

5C18-MS-II 5PYENAP

O

CH3

H3C

R1

R2H3C H

HO

H3C H

CH3

CH3

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Excellent separation for structural isomers

Selectivity for Dipole-dipole Interactions1,8-dinitronaphthalene because of the strong dipole formed by the two nitro groups positioned on

the same side of naphthalene.

Attention1.

-

2.

from silica gel and elutes, even a slight quantity can be detected and causes baseline noise. In such a case, wash the column

with tetrahydrofuran. Detachment of a small amount of the stationary phase does not deteriorate a columns separation ability.

3.

Ordering InformationAnalytical / Preparative Column

NACALAI TESQUE, INC

Column size: 4.6mmI.D.-150mmMobile phase: NPE Methanol / H2O = 70/30

PYE Methanol / H2O = 90/10C18-MS-II Methanol / H2O = 80/20

Flow rate: 1.0 ml/min Temperature: 30 CDetection: UV254nmSample: 1; 1,8-Dinitronaphthalene

2; 1,5-Dinitronaphthalene3; Naphthalene4; 1-Methylnaphthalene5; 1,5-Dimethylnaphthalene

AP-1074

5NPE 5C18-MS-II5PYE

CH3

CH3

NO2 NO2 NO2

NO2

CH3

1 2 3 4 5

Comparison of Separation PropertyComparison of Separation Property


1.0 x 150 05897-012.0 x 150 34328-512.0 x 250 34379-91


4.6 x 150 37902-214.6 x 250 37990-7110 x 250 05469-1120 x 250 38046-21


4.6 x 10 37904-0110 x 20 38045-3120 x 20 05868-8120 x 50 05869-71

Documents

HPLC Column for Structual Isomers - obrnutafaza.hr · Column Size Mobile Phase Flow Rate Temperature Detection 4.6 mm I.D. x 150 mm PYE: Methanol : Water = 90 : 10 NPE: Methanol :