HPLC Method Development Guidelines - Phenomenexphx.phenomenex.com/lib/PO97080313_W.pdf · Xylene...
1
PO97080313_W USP: L11 MOBILE PHASE COMPOSITION: BUFFER AND ORGANIC SOLVENTS Henderson-Hasselbalch By using a pH value which is at least two units above or below the pK a value of analyte you ensure that your com- pound is either fully charged or uncharged. Reminder: A buffer always consists of an acid or base and the corresponding salt. Typical concentrations are usually in the range of 10-50 mM depending on the needed ionic strength and solubility of the buffer system. See below for more buffer details. Buffer, pK s and pH Calculated according to the Henderson-Hasselbalch dissociation constant (K a ) [H + ][A - ] [HA] K a = [HA] [A - ] pH = pK a -log [A - ]/[HA] pH 100:1 pK a + 2 10:1 pK a + 1 1:1 pK a 1:10 pK a - 1 1:100 pK a - 2 NH 2 SO 3 SILICA BASE Van der Waals Forces Hydrophobic / Non-polar / Reversed Phase Typical Phases: C18, C8, Phenyl, Polymer (i.e. SDVB) Interactions with alkyl chains Hydrophilic / Polar / Normal Phase Typical Phases: Silica, NH 2 , CN Interactions with silanols and polar functional groups Bond Energies Involved with Retention Mechanisms Physical Interaction Types of Compounds Energy (kJ/mol) Van der Waals Forces Alkyl chains, aromatic rings 2 – 10 Dipole-Dipole Interactions Carbonyls, heterocyclic 8 – 15 Hydrogen Bond Alcohol, amine, acid 20 – 50 Coulomb Forces Ionic compounds 100 – 400 Covalent Bond Nonmetals > 300 REVERSED PHASE HPLC METHOD DEVELOPMENT PARAMETERS • Analyte structure and pK a • Mobile phase composition • HPLC column choice (optimal stationary phase) RETENTION MECHANISMS HPLC Method Development Guidelines Hydrogen bonding and dipole-dipole interactions O C N H Si O O O O H H OH SILICA BASE CO 2 SILICA BASE SO 3 OCON(CH 3 ) 2 NH 3 + + N CO 2 H OH Electrostatic and Ionic Interactions Ion-Exchange Typical Phases: SCX, WCX, SAX, NH 2 Interactions with ionic functional groups and strong/weak ion-exchangers Buffer Selection Buffer * pK a Buffer Range (pH) MS Compatibility Trifluoroacetic Acid < 2 < 2.5 • ** Phosphoric Acid (pK 1 ) 2.1 1.5 - 2.7 Citric Acid (pK 1 ) 3.1 2.5 - 3.7 Formic Acid 3.8 2.8 - 4.8 • Citrate (pK 2 ) 4.7 4.1 - 5.3 Acetic Acid 4.8 3.8 - 5.8 • Citrate (pK 3 ) 5.4 4.8 - 6.0 Carbonate (pK 1 ) 6.4 5.4 - 7.4 • Phosphate (pK 2 ) 7.2 6.6 - 7.8 Triethanolamine 7.8 6.8 - 8.8 • TRIS 8.3 7.3 - 9.3 Diethanolamine 8.9 7.9 - 9.9 • Ammonia 9.2 8.2 - 10.2 • Ethanolamine 9.5 8.5 - 10.5 • Carbonate (pK 2 ) 10.3 9.3 - 11.3 • Diethylamine 10.5 9.5 - 11.5 • Triethylamine 11.0 10.0 - 12.0 • Piperidin 11.1 10.1 - 12.1 Sources: Practical HPLC Method Development; L.R. Snyder, JJ Kirkland, and JL Glajch. Wiley Interscience, 1997. Introduction to Protein and Peptide HPLC; TP Bradshaw, Phenomenex, 1998. * Common buffers are in bold ** Low concentrations of TFA may be used in LC/MS applications, however it can affect sensitivity. Miscibility Table *Immiscible means that in some proportions two phases will be produced. Immiscible* Xylene Water Trichloroethylene Toluene Tetrahydrofuran di-iso-Propyl Ether iso-propanol 6 n-Propanol Pentane Methyl Ethyl Ketone 5 Methyl-t-Butyl Ether 4 Methanol Hexane Heptane di-Ethyl Ether Ethyl Acetate Ethanol Dioxane Dimethyl Sulfoxide 3 Dimethylformamide Dichlormethane 2 1,2-Dichloroethane 1 Cyclohexane Chloroform Carbon tetrachloride n-Butanol Butyl Acetate Benzene Acetonitrile Acetone Acetic Acid & Miscible SYNONYM TABLE 1 Ethylene Chloride 2 Methylene Chloride 3 Methyl Sulfoxide 4 tert-Butyl Methyl Ether 5 2-Butanone 6 2-Propanol Polarity Refractive UV (nm) Boiling Point Viscosity Solubility in Solvent Index Index @ 20 °C Cutoff @ 1 AU (°C) (cPoise) Water (% w/w) Acetic Acid 6.2 1,372 230 118 1.26 100 Acetone 5.1 1,359 330 56 0.32 100 Acetonitrile 5.8 1,344 190 82 0.37 100 Benzene 2.7 1,501 280 80 0.65 0.18 Butyl Acetate 4.0 1,394 254 125 0.73 0.43 n-Butanol 3.9 1,399 215 118 2.98 7.81 Carbon tetrachloride 1.6 1,466 263 77 0.97 0.08 Chloroform 4.1 1,446 245 61 0.57 0.815 Cyclohexane 0.2 1,426 200 81 1.00 0.01 1,2-Dichloroethane 1 3.5 1,444 225 84 0.79 0.81 Dichloromethane 2 3.1 1,424 235 41 0.44 1.6 Dimethylformamide 6.4 1,431 268 155 0.92 100 Dimethyl Solfoxide 3 7.2 1,478 268 189 2.00 100 Dioxane 4.8 1,422 215 101 1.54 100 Ethyl Acetate 4.4 1,372 260 77 0.45 8.7 Ethanol 5.2 1,360 210 78 1.20 100 di-Ethyl Ether 2.8 1,353 220 35 0.32 6.89 Heptane 0.0 1,387 200 98 0.39 0.0003 Hexane 0.0 1,375 200 69 0.33 0.001 Methanol 5.1 1,329 205 65 0.60 100 Methyl-t-Butyl Ether 4 2.5 1,369 210 55 0.27 4.8 Methyl Ethyl Ketone 5 4,7 1,379 329 80 0.45 24 Pentane 0.0 1,358 200 36 0.23 0.004 n-Propanol 4.0 1,384 210 97 2.27 100 iso-Propanol 6 3.9 1,377 210 82 2.30 100 di-iso-Propyl Ether 2.2 1,368 220 68 0.37 Tetrahydrofuran 4.0 1,407 215 65 0.55 100 Toluene 2.4 1,496 285 111 0.59 0.051 Trichloroethylene 1.0 1,477 273 87 0.57 0.11 Water 9.0 1,333 200 100 1.00 100 Xylene 2.5 1,500 290 139 0.61 0.018 Solvent Strength WEAK Water Methanol Iso-propyl Alcohol Acetonitrile Acetone Ethyl Acetate Ethyl Ether Tetrahydrofuran Methylene Chloride Chloroform Toluene Iso-octane Hexane STRONG TIP: For aromatic compounds, phenyl phases offer a different selectivity then alkyl phases (C18, C8). Based on the interactions between the pi-electrons of the stationary phase‘s phenyl ring and the double bonds in the aromatic ring(s) of your analyte, retention time and elution order can change signifi- cantly and thus have a very positive effect on your method. You can further adjust the selectivity of a phenyl stationary phase by using methanol instead of acetonitrile within the mobile phase. The pi-electrons of acetonitrile can interact with both the phenyl phase and aromatic func- tional groups in a competing way, thus leading to a reduced analyte reten- tion time. TIP: Use both methanol and acetonitrile to vary the selectivity. Pentafluorophenyl phase offers a high degree of steric interac- tions for improved separation of structural isomers, and the electronegative fluorine groups can offer increased retention of polar basic compounds TMS TMS TMS TMS XB- C18 Si TMS TMS TMS TMS HILIC O O OH H20 H20 H20 H20 H20 H20 H20 H20 H20 H20 H20 H20 ACN ACN ACN ACN ACN ACN ACN ACN ACN ACN ACN ACN ACN ACN ACN ACN HO HO OH Si Si Si HO OH TMS F F F F F TMS ACHIEVE THE BEST RESOLUTION with proper selectivity Aeris ™ Core-Shell Phases Use the Aeris PEPTIDE for the peptides and proteins up to 10 kDa and use the Aeris WIDEPORE for proteins larger than 10 kDa • Aeris PEPTIDE and WIDEPORE • Hydrophilic peptides and proteins • PEGylated proteins • High temperature separations • Alternate selectivity for peptide mapping • Aeris WIDEPORE • Large proteins • Moderately hydrophobic proteins • Monoclonal antibodies • High temperature separations • Aeris WIDEPORE • Very large proteins • Very hydrophobic proteins • Membrane proteins • Least retentive Si TMS TMS XB- C18 Si TMS TMS TMS TMS Kinetex ® Core-Shell Phases Use these different selectivities for the analysis of small compounds up to 10 kDa This unique C18 phase yields increased hydrogen bonding with hydrophobic selectivity, resulting in improved peak shape for basic compounds and increased retention of acidic compounds Balanced C18 phase that provides the highest degree of hydrophobic selectivity relative to the other Kinetex phases Used under HILIC running conditions, this phase provides the highest polar selectivity for retention and separation of hydrophilic compounds Aromatic and moderate hydro- phobic selectivity result in the great retention and separation of aromatic hydrocarbons Moderate hydrophobic and steric selectivity is offered, bringing ultrahigh performance to USP L7 and other octyl silane methods Trademarks Kinetex is a registered trademark and Aeris is a trademark of Phenomenex. © 2013 Phenomenex, Inc. All rights reserved. Don’t hesitate to contact us with any questions! Web: www.phenomenex.com / Email: [email protected] Core-Shell Technology for Proteins and Peptides Core-Shell Technology Performance gains on ANY LC System Learn more at: www.phenomenex.com/Kinetex Learn more at: www.phenomenex.com/Aeris PERFORMANCE GAINS ON ANY LC SYSTEM USP: L1 USP: L1 USP: L7 Phases Ligand Description Selectivity Profile Si TMS TMS Kinetex XB-C18 Di-isobutyl side chains differentiate this C18 column. Low ligand density and an inactive surface make this column a great hydrogen acceptor. This phase will demonstrate improved peak shape for basic compounds and increased retention of acids. Cation Selectivity at pH 7.0 Cation Selectivity at pH 2.8 Hydrogen Bond Accepting Capacity Hydrogen Bond Donating Capacity Steric Interaction Hydrophobicity Low High TMS TMS Kinetex C18 Very well balanced column providing some selectivity through steric, hydrogen, and cationic pathways. This is a great starting point for ultra-high efficiency separations. Cation Selectivity at pH 7.0 Cation Selectivity at pH 2.8 Hydrogen Bond Accepting Capacity Hydrogen Bond Donating Capacity Steric Interaction Hydrophobicity Low High TMS TMS Kinetex C8 Brings the benefits of core-shell technology to USP L7 methods. The phase will provide moderate hydrophobicity and good steric and hydrogen donating selectivity. Cation Selectivity at pH 7.0 Cation Selectivity at pH 2.8 Hydrogen Bond Accepting Capacity Hydrogen Bond Donating Capacity Steric Interaction Hydrophobicity Low High TMS F F F F F TMS Kinetex PFP This pentafluorophenyl column provides a very high degree of steric selectivity to separate structural isomers. The electronegative fluorine groups offer high selectivity for cationic compounds. Cation Selectivity at pH 7.0 Cation Selectivity at pH 2.8 Hydrogen Bond Accepting Capacity Hydrogen Bond Donating Capacity Steric Interaction Hydrophobicity Low High Kinetex Phenyl-Hexyl Aromatic and moderate hydrophobic selectivity result in the great retention and separation of aromatic hydrocarbons. TMS TMS USP: L11 Cation Selectivity at pH 7.0 Cation Selectivity at pH 2.8 Hydrogen Bond Accepting Capacity Hydrogen Bond Donating Capacity Steric Interaction Hydrophobicity Low High Columns are pH stable from 1.5-10 under isocratic conditions. Columns are pH stable 1.5-8.5 under gradient conditions. 2.1 mm ID Kinetex columns are pressure stable up to 1000 bar. When using Kinetex 1.3 μm or 1.7 μm, increased performance can be achieved, however high pressure-capable instrumentation is required. ™ ™ 5 µm ™ 2,6 µm ™ 1,7 µm ™ 1,3 µm WIDEPORE PEPTIDE USP: L43 USP: L7 USP: L1 USP: L1
Transcript of HPLC Method Development Guidelines - Phenomenexphx.phenomenex.com/lib/PO97080313_W.pdf · Xylene...
PO
9708
0313
_W
USP: L11
Mobile Phase CoMPosition: buffer and organiC solvents Henderson-HasselbalchBy using a pH value which is at least two units above or below the pKa value of analyte you ensure that your com-pound is either fully charged or uncharged.Reminder: A buffer always consists of an acid or base and the corresponding salt. Typical concentrations are usually in the range of 10-50 mM depending on the needed ionic strength and solubility of the buffer system. See below for more buffer details.
Buffer, pKs and pHCalculated according to the Henderson-Hasselbalchdissociation constant (Ka)
[H+][A-][HA]Ka=
[HA][A-]pH = pKa-log
[A-]/[HA] pH
100:1 pKa + 2
10:1 pKa + 1
1:1 pKa
1:10 pKa - 1
1:100 pKa - 2
NH2
SO3
SILIC
A B
AS
E
Van der Waals Forces
Hydrophobic / Non-polar / Reversed PhaseTypical Phases: C18, C8, Phenyl, Polymer (i.e. SDVB)
Interactions with alkyl chains
Hydrophilic / Polar / Normal PhaseTypical Phases: Silica, NH2, CN
Interactions with silanols and polar functional groups
Bond Energies Involved with Retention MechanismsPhysical Interaction Types of Compounds Energy
(kJ/mol)
Van der Waals Forces Alkyl chains, aromatic rings
2 – 10
Dipole-Dipole Interactions Carbonyls, heterocyclic 8 – 15
Hydrogen Bond Alcohol, amine, acid 20 – 50
Coulomb Forces Ionic compounds 100 – 400
Covalent Bond Nonmetals > 300
reversed Phase hPlC Method develoPMent ParaMeters• Analyte structure and pKa
• Mobile phase composition
• HPLC column choice (optimal stationary phase)
retention MeChanisMs
HPLC Method Development Guidelines
Hydrogen bonding and dipole-dipole interactions
O
CN
H
SiO
O O O
H
H
OH
SILIC
A B
AS
E
CO2
SILIC
A B
AS
E
SO3
OCON(CH3)2
NH3+
+N
CO2H
OH
Electrostatic and Ionic Interactions
Ion-ExchangeTypical Phases: SCX, WCX, SAX, NH2
Interactions with ionic functional groups and strong/weak ion-exchangers
Buffer Selection
Buffer* pKa
Buffer Range (pH)
MS Compatibility
Trifluoroacetic Acid < 2 < 2.5 •**
Phosphoric Acid (pK1) 2.1 1.5 - 2.7Citric Acid (pK1) 3.1 2.5 - 3.7Formic Acid 3.8 2.8 - 4.8 •Citrate (pK2) 4.7 4.1 - 5.3Acetic Acid 4.8 3.8 - 5.8 •Citrate (pK3) 5.4 4.8 - 6.0Carbonate (pK1) 6.4 5.4 - 7.4 •Phosphate (pK2) 7.2 6.6 - 7.8Triethanolamine 7.8 6.8 - 8.8 •TRIS 8.3 7.3 - 9.3Diethanolamine 8.9 7.9 - 9.9 •Ammonia 9.2 8.2 - 10.2 •Ethanolamine 9.5 8.5 - 10.5 •Carbonate (pK2) 10.3 9.3 - 11.3 •Diethylamine 10.5 9.5 - 11.5 •Triethylamine 11.0 10.0 - 12.0 •Piperidin 11.1 10.1 - 12.1
Sources: Practical HPLC Method Development; L.R. Snyder, JJ Kirkland, and JL Glajch. Wiley Interscience, 1997. Introduction to Protein and Peptide HPLC; TP Bradshaw, Phenomenex, 1998. * Common buffers are in bold ** Low concentrations of TFA may be used in LC/MS applications, however it can affect sensitivity.
Miscibility Table
*Immiscible means that in some proportions two phases will be produced.
Immiscible*
Xylen
eWa
terTri
chlor
oeth
ylene
Tolue
neTe
trahy
drofu
ran
di-iso
-Pro
pyl E
ther
iso-p
ropa
nol6
n-Pr
opan
olPe
ntan
eMe
thyl
Ethy
l Keto
ne5
Meth
yl-t-B
utyl
Ethe
r4
Meth
anol
Hexa
neHe
ptan
edi-
Ethy
l Eth
erEt
hyl A
cetat
eEt
hano
lDi
oxan
eDi
meth
yl Su
lfoxid
e3
Dim
ethylf
orm
amide
Dich
lorm
ethan
e2
1,2-
Dich
loroe
than
e1
Cyclo
hexa
neCh
lorofo
rmCa
rbon
tetra
chlor
iden-
Butan
olBu
tyl Ac
etate
Benz
ene
Aceto
nitrile
Aceto
neAc
etic A
cid
& Miscible SYNONYM TABLE1 Ethylene Chloride2 Methylene Chloride3 Methyl Sulfoxide
4 tert-Butyl Methyl Ether5 2-Butanone6 2-Propanol
Polarity Refractive UV (nm) Boiling Point Viscosity Solubility inSolvent Index Index @ 20 °C Cutoff @ 1 AU (°C) (cPoise) Water (% w/w)
Acetic Acid 6.2 1,372 230 118 1.26 100Acetone 5.1 1,359 330 56 0.32 100Acetonitrile 5.8 1,344 190 82 0.37 100Benzene 2.7 1,501 280 80 0.65 0.18Butyl Acetate 4.0 1,394 254 125 0.73 0.43n-Butanol 3.9 1,399 215 118 2.98 7.81Carbon tetrachloride 1.6 1,466 263 77 0.97 0.08Chloroform 4.1 1,446 245 61 0.57 0.815Cyclohexane 0.2 1,426 200 81 1.00 0.011,2-Dichloroethane1 3.5 1,444 225 84 0.79 0.81Dichloromethane2 3.1 1,424 235 41 0.44 1.6Dimethylformamide 6.4 1,431 268 155 0.92 100Dimethyl Solfoxide3 7.2 1,478 268 189 2.00 100Dioxane 4.8 1,422 215 101 1.54 100Ethyl Acetate 4.4 1,372 260 77 0.45 8.7Ethanol 5.2 1,360 210 78 1.20 100di-Ethyl Ether 2.8 1,353 220 35 0.32 6.89Heptane 0.0 1,387 200 98 0.39 0.0003Hexane 0.0 1,375 200 69 0.33 0.001Methanol 5.1 1,329 205 65 0.60 100Methyl-t-Butyl Ether4 2.5 1,369 210 55 0.27 4.8Methyl Ethyl Ketone5 4,7 1,379 329 80 0.45 24Pentane 0.0 1,358 200 36 0.23 0.004n-Propanol 4.0 1,384 210 97 2.27 100iso-Propanol6 3.9 1,377 210 82 2.30 100di-iso-Propyl Ether 2.2 1,368 220 68 0.37 Tetrahydrofuran 4.0 1,407 215 65 0.55 100Toluene 2.4 1,496 285 111 0.59 0.051Trichloroethylene 1.0 1,477 273 87 0.57 0.11Water 9.0 1,333 200 100 1.00 100Xylene 2.5 1,500 290 139 0.61 0.018
Solvent Strength wEAk
Water
Methanol
Iso-propyl Alcohol
Acetonitrile
Acetone
Ethyl Acetate
Ethyl Ether
Tetrahydrofuran
Methylene Chloride
Chloroform
Toluene
Iso-octane
Hexane
STRONG
TIP: For aromatic compounds, phenyl phases offer a different selectivity then alkyl phases (C18, C8). Based on the interactions between the pi-electrons of the stationary phase‘s phenyl ring and the double bonds in the aromatic ring(s) of your analyte, retention time and elution order can change signifi-cantly and thus have a very positive effect on your method.
You can further adjust the selectivity of a phenyl stationary phase by using methanol instead of acetonitrile within the mobile phase. The pi-electrons of acetonitrile can interact with both the phenyl phase and aromatic func-tional groups in a competing way, thus leading to a reduced analyte reten-tion time.
TIP: Use both methanol and acetonitrile to vary the selectivity.
Pentafluorophenyl phase offers a high degree of steric interac-tions for improved separation of structural isomers, and the electronegative fluorine groups can offer increased retention of polar basic compounds
C8
TMSTMS
C18
TMSTMS
XB-C18
Si TMSTMS
Phenyl-Hexyl
TMSTMS
HILIC
OO
OHH20
H20H20 H20
H20
H20
H20
H20 H20
H20
H20H20
ACN ACN ACN ACN
ACN ACN ACN ACN
ACN ACN ACN ACN
ACN ACN ACN ACN
HO
HO
OHSi
SiSi
HO
OH
PFP
TMS
F
F
F
F
F
TMS
aChieve the best resolution with proper selectivity
Aeris™ Core-Shell PhasesUse the Aeris PEPTIDE for the peptides and proteins up to 10 kDa and use the Aeris WIDEPORE for proteins larger than 10 kDa
• Aeris PEPTIDE and WIDEPORE
• Hydrophilic peptides and proteins
• PEGylated proteins
• High temperature separations
• Alternate selectivity for peptide mapping
• Aeris WIDEPORE
• Large proteins
• Moderately hydrophobic proteins
• Monoclonal antibodies
• High temperature separations
• Aeris WIDEPORE
• Very large proteins
• Very hydrophobic proteins
• Membrane proteins
• Least retentiveXB-C8
Si TMSTMS
XB-C18
Si TMSTMS
C4
TMSTMS
kinetex® Core-Shell PhasesUse these different selectivities for the analysis of small compounds up to 10 kDa
This unique C18 phase yields increased hydrogen bonding with hydrophobic selectivity, resulting in improved peak shape for basic compounds and increased retention of acidic compounds
Balanced C18 phase that provides the highest degree of hydrophobic selectivity relative to the other Kinetex phases
Used under HILIC running conditions, this phase provides the highest polar selectivity for retention and separation of hydrophilic compounds
Aromatic and moderate hydro-phobic selectivity result in the great retention and separation of aromatic hydrocarbons
Moderate hydrophobic and steric selectivity is offered, bringing ultrahigh performance to USP L7 and other octyl silane methods
Trademarks Kinetex is a registered trademark and Aeris is a trademark of Phenomenex. © 2013 Phenomenex, Inc. All rights reserved.
Don’t hesitate to contact us with any questions! Web: www.phenomenex.com / Email: [email protected]
Core-Shell Technology for Proteins and Peptides
Core-Shell Technology Performance gains on ANY LC System
Learn more at: www.phenomenex.com/Kinetex
Learn more at: www.phenomenex.com/aeris
PerforManCe gains on anY lC sYsteM
USP: L1
USP: L1
USP: L7
Phases
Ligand Description Selectivity Profile
XB-C18
Si TMSTMS
Kinetex Xb-C18Di-isobutyl side chains differentiate this C18 column. Low ligand density and an inactive surface make this column a great hydrogen acceptor. This phase will demonstrate improved peak shape for basic compounds and increased retention of acids.
Cation Selectivity at pH 7.0Cation Selectivity at pH 2.8
Hydrogen Bond Accepting CapacityHydrogen Bond Donating Capacity
Steric InteractionHydrophobicity
Low High
TMSTMSKinetex C18Very well balanced column providing some selectivity through steric, hydrogen, and cationic pathways. This is a great starting point for ultra-high efficiency separations.
Cation Selectivity at pH 7.0Cation Selectivity at pH 2.8
Hydrogen Bond Accepting CapacityHydrogen Bond Donating Capacity
Steric InteractionHydrophobicity
Low High
TMSTMSKinetex C8Brings the benefits of core-shell technology to USP L7 methods. The phase will provide moderate hydrophobicity and good steric and hydrogen donating selectivity.
Cation Selectivity at pH 7.0Cation Selectivity at pH 2.8
Hydrogen Bond Accepting CapacityHydrogen Bond Donating Capacity
Steric InteractionHydrophobicity
Low High
TMS
F
F
F
F
F
TMSKinetex PfPThis pentafluorophenyl column provides a very high degree of steric selectivity to separate structural isomers. The electronegative fluorine groups offer high selectivity for cationic compounds.
Cation Selectivity at pH 7.0Cation Selectivity at pH 2.8
Hydrogen Bond Accepting CapacityHydrogen Bond Donating Capacity
Steric InteractionHydrophobicity
Low High
Kinetex Phenyl-hexylAromatic and moderate hydrophobic selectivity result in the great retention and separation of aromatic hydrocarbons.
TMSTMS
USP: L11
Cation Selectivity at pH 7.0Cation Selectivity at pH 2.8
Hydrogen Bond Accepting CapacityHydrogen Bond Donating Capacity
Steric InteractionHydrophobicity
Low High
Columns are pH stable from 1.5-10 under isocratic conditions. Columns are pH stable 1.5-8.5 under gradient conditions.
2.1 mm ID Kinetex columns are pressure stable up to 1000 bar. When using Kinetex 1.3 μm or 1.7 μm, increased performance can be achieved, however high pressure-capable instrumentation is required.
™
™
™
5 µm
™
2,6 µm
™
1,7 µm
™
1,3 µm
WIDEPORE PEPTIDE
USP: L43
USP: L7
USP: L1
USP: L1
