Pentabromobenzyl Group Bonded HPLC Column COSMOSIL PBr · COSMOSIL PBr shows excellent separation...
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• Separate hydrophilic compounds in reversed-phase conditions • Simple mobile phase condition compared to HILIC • More sample loading capacity than HILIC • Alternative selectivity to C18 column Separation of hydrophilic compounds in reversed-phase conditions Hydrophilic interaction chromatography (HILIC) is an increasingly popular analytical technique. However, it is often difficult to develop a robust method due to users’ less familiarity with HILIC conditions. Furthermore, high concentration of acetonitrile used in HILIC mobile phase makes it extremely sensitive to samples’ water concentration; injecting samples in high water concentration often results in poor peak shapes. COSMOSIL PBr enables separation of hydrophilic compounds in reversed- phase conditions, maintaining sharp peak shapes even with aqueous samples. COSMOSIL Application Data COSMOSIL Application Data NACALAI TESQUE, INC Column: HILIC Column size: 4.6mmI.D.-150mm Mobile phase: Acetonitrile/ H 2 O = 90/10 Flow rate: 1.0 ml/min Temperature: 30°C Detection: UV260nm Sample: 1; Uracil (0.1mg/ml) 2; Uridine (0.2mg/ml) Sam. Solution H 2 O 150mm Column: PBr Column size: 4.6mmI.D.- Mobile phase: H 2 O Flow rate: 1.0 ml/min Temperature: 30°C Detection: UV260nm Sample: 1; Uracil (0.1mg/ml) 2; Uridine (0.2mg/ml) Sam. Solution H 2 O Undiluted sol. 1μl Injection 1/20 dilute sol. 20μl Injection Undiluted sol. 1μl Injection 1/20 dilute sol. 20μl Injection N H NH O O NH O O N O OH OH H H H H HO 1; Uracil 2; Uridine Comparison with C18 Columns COSMOSIL PBr retains hydrophilic compounds stronger than C18 columns under the same reversed-phase conditions. COSMOSIL Application Data COSMOSIL Application Data NACALAI TESQUE, INC 5C 18 -PAQ PBr Column: Column size: 4.6mmI.D.-250mm Mobile phase: 20mmol/l Phosphate buffer(pH7.0) Flow rate: 1.0 ml/min Temperature: 30°C Detection: UV225nm Sample: 1; Ammelide (0.1mg/ml) 2; Cyanuric Acid (0.5mg/ml) 3; Ammeline (0.1mg/ml) 4; Melamine (0.1mg/ml) Inj.Vol.: 1.0μl N N N NH2 HO OH HN N H NH O O O N N N NH2 HO NH2 N N N NH2 H2N NH2 1; Ammelide 2; Cyanuric Acid 3; Ammeline 4; Melamine T 0 ; Retention time of unretained peak Pentabromobenzyl Group Bonded HPLC Column COSMOSIL PBr
Transcript of Pentabromobenzyl Group Bonded HPLC Column COSMOSIL PBr · COSMOSIL PBr shows excellent separation...
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• Separatehydrophiliccompoundsinreversed-phaseconditions• SimplemobilephaseconditioncomparedtoHILIC• MoresampleloadingcapacitythanHILIC• AlternativeselectivitytoC18column
Separation of hydrophilic compounds in reversed-phase conditions Hydrophilic interaction chromatography (HILIC) is an increasingly popular analytical technique. However, it is often difficult to develop a robust method due to users’ less familiarity with HILIC conditions. Furthermore, high concentration of acetonitrile used in HILIC mobile phase makes it extremely sensitive to samples’ water concentration; injecting samples in high water concentration often results in poor peak shapes. COSMOSIL PBr enables separation of hydrophilic compounds in reversed-phase conditions, maintaining sharp peak shapes even with aqueous samples.
COSMOSIL Application DataCOSMOSIL Application Data
NACALAI TESQUE, INC
Column: HILIC Column size: 4.6mmI.D.-150mmMobile phase: Acetonitrile/ H2O = 90/10Flow rate: 1.0 ml/min Temperature: 30°CDetection: UV260nm
Sample: 1; Uracil (0.1mg/ml)2; Uridine (0.2mg/ml)
Sam. Solution H2O
150mmColumn: PBr Column size: 4.6mmI.D.-Mobile phase: H2OFlow rate: 1.0 ml/min Temperature: 30°CDetection: UV260nm
Sample: 1; Uracil (0.1mg/ml)2; Uridine (0.2mg/ml)
Sam. Solution H2O
Undiluted sol.
1µl Injection
1/20 dilute sol.
20µl Injection
Undiluted sol.
1µl Injection
1/20 dilute sol.
20µl Injection
NH
NH
O
O
NH
O
ON
O
OHOH
HHHH
HO
1; Uracil 2; Uridine
Comparison with C18 ColumnsCOSMOSIL PBr retains hydrophilic compounds stronger than C18 columns under the same reversed-phase conditions.
COSMOSIL Application DataCOSMOSIL Application Data
NACALAI TESQUE, INC
5C18-PAQ PBr
Column: Column size: 4.6mmI.D.-250mmMobile phase: 20mmol/l Phosphate buffer(pH7.0)Flow rate: 1.0 ml/min Temperature: 30°CDetection: UV225nm
Sample: 1; Ammelide (0.1mg/ml)2; Cyanuric Acid (0.5mg/ml)3; Ammeline (0.1mg/ml)4; Melamine (0.1mg/ml)
Inj.Vol.: 1.0µl
N
N
N
NH2HO
OHHN
NH
NH
O
OON
N
N
NH2HO
NH2
N
N
N
NH2H2N
NH2
1; Ammelide 2; Cyanuric Acid 3; Ammeline 4; Melamine T0; Retention time of unretained peak
Pentabromobenzyl Group Bonded HPLC Column
COSMOSIL PBr
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Separation MechanismIn reversed-phase chromatography, compounds are separated by difference in hydrophobicity (the number of CH2 bases, see chromatogram on the left (1)). Compounds with little hydrophobicity (the number of OCH2CH2, see chromatogram on the right (2)), are not retained by a C18 column. However, these compounds can easily be separated by dispersion force interaction using the COSMOSIL PBr column.
NACALAI TESQUE, INC
Column: Column size: 4.6mmI.D.-150mmMobile phase: Methanol/ H2O = 80/20Flow rate: 1.0 ml/min Temperature: 30°CDetection: UV254nm
Sample: 1; Benzene2; Toluene3; Ethylbenzene4; n-Propylbenzene5; n-Butylbenzene
Column: Column size: 4.6mmI.D.-150mmMobile phase: Methanol/ H2O = 60/40Flow rate: 1.0 ml/min Temperature: 30°CDetection: UV254nm
Sample: 1; Benzyl Alcohol2; Ethylene Glycol Monobenzyl Ether3; Diethylene Glycol Monobenzyl Ether4; Triethylene Glycol Monobenzyl Ether5; Tetraethylene Glycol Monobenzyl Ether
(CH2)n-H
n=0-4
5C18-MS-II PBr5C18-MS-II PBr
CH2 (OCH2CH2)n-OH
n=0-4
(1) Compounds with more hydrophobicity (2) Compounds with less hydrophobicity
Dispersion force effect
► Dispersion force (instantaneous dipole–induced dipole force)
London Dispersion force is a weak intermolecular force that results from dipoles temporarily induced from random unsymmetrical electron positions in two adjacent atoms, also known as “instantaneous dipole–induced dipole force”. It is present in all molecules regardless if they are polar or non-polar. Compounds with high polarizability have stronger dispersion force.
► Compounds with stronger dispersion force
• Larger and heavier molecules• Molecules with larger and heavier atoms (e.g. from weakest to strongest in halogens, F2, Cl2, Br2, and
I2)• Molecules with delocalized electrons and resonance (e.g. aromatic compounds)
COSMOSIL PBr column is packed with pentabromobenzyl bonded silica that enables separation by dispersion force interaction.
Suitable Compounds to Analyze by COSMOSIL PBrCOSMOSIL PBr retains cyclic compounds or amide compounds more strongly than C18 columns.
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Dop
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Am
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ide
Cya
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mm
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Nic
otin
amid
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tam
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1A
cryl
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cryl
amid
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MS
OD
MF
C18-PAQ
PBr
Ret
entio
n ra
tio
of e
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sam
ple
(k’
)
Nucleic aci
d bases
(1) Catechola
mines
(2) Organi
c acids
(2) Amino a
cids
(2)Melam
ine
related ma
terials
(3)Wate
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vitamins
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(4)Solve
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(2)
(Mobile phase)(1) Methanol/ 20mmol/l Phosphate buffer(pH7.0) = 10/90(2) 20mmol/l Phosphate buffer(pH2.5)(3) 20mmol/l Phosphate buffer(pH7.0)(4) Methanol/ H2O = 10/90
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ApplicationsGentamycin Nucleic Acid Base
COSMOSIL Application DataCOSMOSIL Application Data
NACALAI TESQUE, INC
Column: PBrColumn size: 2.0mmI.D.-150mmMobile phase: 0.1% Pentafluoropropionic Acid
-Acetonitrile/ H2O = 10/90Flow rate: 0.2 ml/min Temperature: 40°CDetection: ESI-MS, Positive, SIM
Sample: Gentamicin (C1a, C2, C2a, C1)
Inj.Vol.: 1.0µl
COSMOSIL Application DataCOSMOSIL Application Data
NACALAI TESQUE, INC
Column: PBrColumn size: 4.6mmI.D.-150mmMobile phase: Methanol/ 20mmol/l Phosphate
buffer(pH7.0) = 10/90Flow rate: 1.0 ml/min Temperature: 30°CDetection: UV260nm
Sample: 1; Cytosine (0.05mg/ml) 2; Uracil (0.05mg/ml) 3; Thymine (0.05mg/ml) 4; Guanine (0.05mg/ml) 5; Adenine (0.05mg/ml)
Inj.Vol.: 1.0µl
Coenzyme A Methylimidazole Isomers
COSMOSIL Application DataCOSMOSIL Application Data
NACALAI TESQUE, INC
Column: PBrColumn size: 4.6mmI.D.-150mmMobile phase: Methanol/ 20mmol/l Phosphate
buffer(pH7.0) = 20/80Flow rate: 1.0 ml/min Temperature: 30°CDetection: UV260nm
Sample: 1; Malonyl Coenzyme A (0.25mg/ml) 2; Coenzyme A (0.25mg/ml) 3; Acetyl Coenzyme A (0.25mg/ml)
Inj.Vol.: 2.0µl
COSMOSIL Application DataCOSMOSIL Application Data
NACALAI TESQUE, INC
Column: PBrColumn size: 4.6mmI.D.-250mmMobile phase: 20mmol/l Phosphate buffer(pH2.5)Flow rate: 1.0 ml/min Temperature: 30°CDetection: UV220nm
Sample: 1; 1-Methylimidazole (0.1mg/ml)2; 2-Methylimidazole (0.1mg/ml)3; 4-Methylimidazole (0.1mg/ml)
Inj.Vol.: 1.0µl
Column Stability TestCOSMOSIL PBr column remains stable after 1000 injections under 100% aqueous condition.
COSMOSIL Application DataCOSMOSIL Application Data
NACALAI TESQUE, INC
#1 #1000
Column: PBr Column size: 4.6mmI.D.-150mmMobile phase: 20mmol/l Phosphate buffer(pH2.5)Flow rate: 1.0 ml/min Temperature: 40°CDetection: UV280nm
Sample: 1; L-Noradrenaline (0.5mg/ml)2; L-Adrenaline (0.5mg/ml)3; Dopamine (0.5mg/ml)4; L-DOPA (0.5mg/ml)
Inj.Vol.: 1.0µl
SpecificationsPacking Material 5PBrSilica gel High-purity porous spherical silicaAverage particle size 5 μmAverage pore size approx. 120ÅSpecific surface area approx. 300 m2/gStationary phase Pentabromobenzyl groupBonding type MonomericEndcapping treatment YesCarbon load approx. 8%Usable pH range 2-7.5
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1505
For research use only, not intended for diagnostic or drug use.
NACALAI TESQUE, INC.Nijo Karasuma, Nakagyo-ku, Kyoto 604-0855 JAPANTELFAXWebsiteE-mail
: +81-(0)75-251-1730: +81-(0)75-251-1763: www.nacalai.com: [email protected]
Applications for Surfactants and Halogen CompoundsSurfactants
COSMOSIL PBr shows excellent separation for surfactants compared to C18 or PFP columns.
COSMOSIL Application DataCOSMOSIL Application Data
NACALAI TESQUE, INC
5C18-MS-II PBr
Column: Column size: 4.6mmI.D.-150mmMobile phase: 5C18-MS-II, 5PFP Methanol/ H2O = 80/20
PBr MethanolFlow rate: 1.0 ml/min Temperature: 30°CDetection: UV254nm
Sample: Triton(R) X-100 (50mg/ml)Inj.Vol.: 2.0µl
5PFP
Halogen Compounds
COSMOSIL PBr shows high selectivity for halogen compounds.
COSMOSIL Application DataCOSMOSIL Application Data
NACALAI TESQUE, INC
5C18-MS-II 5PFP PBr
Column: Column size: 4.6mmI.D.-150mmMobile phase: Methanol/ H2O = 65/35Flow rate: 1.0 ml/min Temperature: 30°CDetection: UV254nm
Sample: 1; Fluorobenzene (1.0mg/ml)2; , , -Trifluorotoluene
Benzotrifluoride (0.25mg/ml)3; Toluene (2.5mg/ml)4; Chlorobenzene (5.0mg/ml)5; Bromobenzene (5.0mg/ml)6; Iodobenzene (2.0mg/ml)
Inj.Vol.: 1.0µl
Ordering Informations
Column SizeI.D. x Length (mm) Product Number
2.0 x 50 12943-612.0 x 100 13245-812.0 x 150 12392-812.0 x 250 13247-613.0 x 50 12592-613.0 x 100 13249-413.0 x 150 13250-013.0 x 250 13251-914.6 x 50 13252-814.6 x 150 12394-614.6 x 250 12395-51
Column SizeI.D. x Length (mm) Product Number
10 x 50 13253-7110 x 100 13254-6110 x 150 13255-5110 x 250 12397-3120 x 50 13257-3120 x 100 13258-2120 x 150 13259-1120 x 250 12398-2128 x 100 13260-7128 x 150 13261-6128 x 250 13262-51
Column SizeI.D. x Length (mm) Product Number
4.6 mm I.D. x 10 mm cartridge* 12444-14
10 x 20 12396-4120 x 20 13256-41
4.6 mm I.D. Cartridge Holder 38009-79
*Cartridge Holder is required.
