Agilent HPLC Column Selection Guide
Agilent HPLC Column Selection Guide Agilent HPLC Column Selection Guide
HPLC Column Selection GuidelinesHPLC Column Selection Flow ChartTo use the column selection guide diagram below, simply follow the path for your analyte and mobile phase. At the far right, follow your finalcolumn selection to the pages indicated.Please see the 2009-2010 Essential Chromatography and Spectroscopy Catalog for a complete listing of LC columns and supplies.Hexane SolubleNormal-Phase Mode with Bare SilicaSee page62Organic SolventSolubleMeOH and MeOH:H 2 O or ACNandACN:H 2 O SolubleNormal-Phase Mode with Bonded PhaseReversed-Phase Mode with Bonded PhaseSee page62See page19THF SolubleGel-Permeation (Small Molecule)Molecular Weight2000Gel-Filtration ChromatographySee page124Water SolubleIon-Exchange Modewith Wide-Pore MaterialSee pages71 & 128Reversed-Phase Modewith Wide-Pore MaterialSee page113Adapted with permission from "Practical HPLC Methodology and Applications," Brian A. Bidlingmeyer, John Wiley & Sons, Inc., New York, p. 1094 Order online at www.agilent.com/chem/store
HPLC Column Selection GuidelinesQuick Guide to ZORBAX Reversed-Phase Bonded PhasesModern ZORBAX RP-HPLC Columns Recommended Uses and Applications Page No.Eclipse PlusAvailable in RRHD and RRHT, 1.8 µm Excellent first choice for method development Long life from pH 2-9 for reliable separations of basic, acidic and neutral compounds Superior peak shape with basic compounds High resolution and efficiency with 1.8, 3.5 and 5 µm columns Rigorous QA/QC testing for greater long-term reproducibility23Poroshell 120Eclipse XDBAvailable in RRHT, 1.8 µmStableBond (SB)Available in RRHD and RRHT, 1.8 µmZORBAX RxAvailable in RRHT, 1.8 µmBonus-RPAvailable in RRHT, 1.8 µmExtend-C18Available in RRHT, 1.8 µm Superficially porous particles for high efficiency at low pressure Sub 2-micron like efficiency with a 2.7 µm particle Endcapped and non-endcapped C18 phases for selectivity optimization Compatible with 400 bar and 600 bar LC's Four selectivity choices for flexible method development High performance over a wide pH range, pH 2-9 Good peak shape for acids, bases and neutrals Long lifetime with extra dense bonding and double endcapping Fast, ultra-fast, and high resolution separations using 1.8 and 3.5 µm columns Choices from capillary to prep Basic, acidic, neutral compounds Exceptional stability at low pH Use of high temperature (up to 90°C for C18, 80°C for C8, C3, Phenyl, CN, and Aq)and low pH as an added selectivity tool Widest selection of bonded phases for different selectivity (C18, C8, C3, CN,Phenyl, Aq) Uses mobile phases for LC/MS with formic acid, acetic acid, or TFA Uses mobile phases with TFA for peptide and protein separation Rapid separations using 1.8 and 3.5 µm columns General separation of basic, acidic and neutral compounds at low pH with differentselectivity than SB columns Rx-C8 is the same as SB-C8 Separating basic compounds in higher aqueous mobile phases General separation of basic, neutral, acidic compounds at mid-range pH or low pH;especially stable at low pH Separating peptides for different selectivity Rapid separations using 3.5 µm columns Separating basic compounds above their pKa in free base form; separation of basic,acidic, neutral compounds at high pH; up to pH 11.5 Uses ammonium hydroxide as mobile phase additive with LC/MS with small moleculesor peptides Separating at high, mid-range and low pH for selectivity changes Rapid separations using 3.5 µm columnsOriginal ZORBAX Columns Recommended Uses and Applications Page No.ZORBAX General separation of basic, acidic, neutral compounds at low pH with differentselectivity than SB columns; higher number of active silanols than SB "Mixed mode" separation at more neutral pH values61ZORBAX ODS Classic(non-endcapped) General separation of basic, acidic, neutral compounds at mid-range to low pHwith different selectivity than SB or XDB columns20324049565261Order online at www.agilent.com/chem/store5
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<strong>HPLC</strong> <strong>Column</strong> <strong>Selection</strong> <strong>Guide</strong>lines<strong>HPLC</strong> <strong>Column</strong> <strong>Selection</strong> Flow ChartTo use the column selection guide diagram below, simply follow the path for your analyte and mobile phase. At the far right, follow your finalcolumn selection to the pages indicated.Please see the 2009-2010 Essential Chromatography and Spectroscopy Catalog for a complete listing of LC columns and supplies.Hexane SolubleNormal-Phase Mode with Bare SilicaSee page62Organic SolventSolubleMeOH and MeOH:H 2 O or ACNandACN:H 2 O SolubleNormal-Phase Mode with Bonded PhaseReversed-Phase Mode with Bonded PhaseSee page62See page19THF SolubleGel-Permeation (Small Molecule)Molecular Weight2000Gel-Filtration ChromatographySee page124Water SolubleIon-Exchange Modewith Wide-Pore MaterialSee pages71 & 128Reversed-Phase Modewith Wide-Pore MaterialSee page113Adapted with permission from "Practical <strong>HPLC</strong> Methodology and Applications," Brian A. Bidlingmeyer, John Wiley & Sons, Inc., New York, p. 1094 Order online at www.agilent.com/chem/store