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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
Column SizeMobile Phase
Flow RateTemperatureDetection
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
Column SizeMobile Phase
Flow RateTemperatureDetection
4.6 mm I.D. x 150 mm5C18-MS-II NAP
5PYE 1.0 ml/min30UV 254 nm
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
Column SizeMobile Phase
Flow RateTemperatureDetection
Sample4.6 mm I.D. x 150 mm5C18-MS-II NAP
5PYE 1.0 ml/min30UV 295 nm
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
4.6 mm I.D. x 150 mmMethanol : Water = 70 : 301.0 ml/min30UV 280 nm
(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)
Column SizeMobile Phase
Flow RateTemperatureDetection
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
4.6 mm I.D. x 150 mmMethanol : Water = 50 : 501.0 ml/min30UV 254 nm
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.
Column SizeMobile Phase
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: info.intl@nacalai.com
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
26
I. H
PLC
Col
umns
II. U
HPL
C C
olum
nsIII
. Pre
para
tive
Pack
ing
Mat
eria
lsIV
. Rel
ated
Pro
duct
sV.
App
licat
ions
VI. T
echn
ical
Not
esVI
I. In
dex
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=
COSMOSIL Application DataCOSMOSIL Application Data
NACALAI TESQUE, INC
Column: Column size: 4.6mmI.D.-150mmMobile phase: 5C18-MS-II Methanol/ H2O = 30/70
NAP Methanol/ H2O = 50/505PYE Methanol/ H2O = 80/20
Flow rate: 1.0 ml/min Temperature: 30 CDetection: UV254nm
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
27
I. HPLC
Colum
nsII. U
HPLC
Colum
nsIV. R
elated ProductsV. A
pplicationsVI. Technical N
otesVII. Index
III. Preparative Packing M
aterials
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
COSMOSIL Application DataCOSMOSIL Application Data
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
COSMOSIL Application DataCOSMOSIL Application Data
NACALAI TESQUE, INC
Column: Column size: 4.6mmI.D.-150mmMobile phase: 5C18-MS-II Methanol/ H2O = 95/5
NAP Methanol/ H2O = 90/105PYE Methanol/ H2O = 90/10
Flow rate: 1.0 ml/min Temperature: 30 CDetection: UV295nm
Sample: -Tocopherol-Tocopherol-Tocopherol-Tocopherol
AP-1028
5C18-MS-II 5PYENAP
O
CH3
H3C
R1
R2H3C H
HO
H3C H
CH3
CH3
28
I. H
PLC
Col
umns
II. U
HPL
C C
olum
nsIII
. Pre
para
tive
Pack
ing
Mat
eria
lsIV
. Rel
ated
Pro
duct
sV.
App
licat
ions
VI. T
echn
ical
Not
esVI
I. In
dex
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
Column SizeI.D. x Length (mm)
1.0 x 150 05897-012.0 x 150 34328-512.0 x 250 34379-91
Column SizeI.D. x Length (mm)
4.6 x 150 37902-214.6 x 250 37990-7110 x 250 05469-1120 x 250 38046-21
Column SizeI.D. x Length (mm)
4.6 x 10 37904-0110 x 20 38045-3120 x 20 05868-8120 x 50 05869-71
Recommended