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  • Agilent ZORBAX ColumnSelection Guide forAnalytical HPLCResolution that Lasts

  • ZORBAX Silica . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2

    Reversed-Phase HPLC Method Development . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4

    ZORBAX Columns for Analytical Chemistry

    ZORBAX StableBond HPLC Columns . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8

    ZORBAX SB-Aq HPLC Columns . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12

    ZORBAX Rx HPLC Columns . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13

    ZORBAX Eclipse XDB HPLC Columns . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14

    ZORBAX Bonus-RP HPLC Columns . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16

    ZORBAX Extend-C18 HPLC Columns . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18

    Additional ZORBAX Analytical HPLC Columns . . . . . . . . . . . . . . . . . . . . . . . . . 20

    ZORBAX High-Throughput HPLC Columns

    ZORBAX CombiHT HPLC Columns . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21

    ZORBAX LC/MS HPLC Columns and Cartridge Columns . . . . . . . . . . . . . . . . 22

    ZORBAX Rapid Resolution 3.5 m HPLC Columns . . . . . . . . . . . . . . . . . . . . . . 23

    High-Throughput Column Ordering HPLC Information . . . . . . . . . . . . . . . . . . 24

    MicroBore and Capillary Columns . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25

    ZORBAX Columns for BioAnalytical Chemistry

    ZORBAX Wide-Pore 300StableBond HPLC Columns . . . . . . . . . . . . . . . . . . . 26

    ZORBAX 300Extend-C18 HPLC Columns . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28

    ZORBAX Eclipse Amino Acid Analysis (AAA) HPLC Columns . . . . . . . . . . . . 30

    ZORBAX Eclipse dsDNA Analysis HPLC Columns . . . . . . . . . . . . . . . . . . . . . . 31

    ZORBAX GF-250/450 Size Exclusion HPLC Columns . . . . . . . . . . . . . . . . . . . . 32

    ZORBAX Oligo HPLC Columns . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33

    Contents

    Standard Fittings

    ZORBAX 15 mm and 30 mm Rapid Resolution cartridges

    75 mm and 150 mmAgilent cartridgecolumns

    CombiHT

  • 2

    ZORBAX Rx-SIL Xerogel(Sol-type) (SIL-type)

    Uniform Sponge-like, Structure Sub Particles Polymeric Network

    Purity High (99.995%) Low-High

    Strength High Moderate

    Pore Size, Particle Size Distribution Narrow Broad

    Pore Size/Surface Area 80/180m2/g 100/300m2/g

    Porosity (%) 50 70

    High pH Resistance Good Poor

    The ZORBAX Rx-SIL process producesultra-pure (99.995%) particles, with very low metal content. The final silicaparticle is fully hydroxylated and of low acidity. The Rx-SIL process alsoallows careful and reproducible controlof pore size and particle size. These key features: purity (low acidity),strength, and careful control of poresize as well as particle size are criticalto excellent chromatographic resultsand are the building blocks of superiorZORBAX bonded-phases.

    Table 1 compares the process used tomake the ZORBAX Rx-SIL particles to a second process the Xerogelprocess commonly used to make silica particles for HPLC columns. To produce silica with the key featuresthat maximizes chromatographic performance purity, strength, con-trolled pore size and particle size distribution, plus higher pH resistance the Agilent ZORBAX process is anexcellent choice.

    Lets look at how these featuresimprove your chromatography.

    Agilent ZORBAX Silica is an EssentialPart of Providing Resolution that Lasts

    Silica-based HPLC columns have longbeen considered the best choice for maximizing resolution with long life-times and good column performance.The advantages of silica include highefficiency, small particle sizes, goodmechanical strength and compatibilitywith a wide range of organic solvents.The quality and consistency of the silica support is critical. Therefore,close attention to the manufacture of the silica particles can result inimproved chromatography for smallbasic molecules as well as peptidesand proteins. Agilent Technologiesmanufactures silica particles using aunique process to provide a superiorsilica support for ZORBAX HPLCcolumns with total control over themanufacturing process.

    The ZORBAX Silica ManufacturingProcess The Making of a Rugged, High-Purity Silica

    All Agilent ZORBAX columns are built from porous silica microspheres(PSM) based on silica sols. The silica particle is made of tiny, solid sol microparticles agglutinated in apatented polymerization process, then fused together at very high temperatures to form the final particle (Figure 1). These strong,durable silica particles are calledZORBAX Rx-SIL and are the base silica for most of the ZORBAXcolumns in this brochure.

    Silica Sol+

    Urea (pH~2)+

    CH2O

    80 pore

    O2 3.5, 5, 7 m

    StableBond SB, 300SBEclipse XDBBonus-RPExtend-C18, 300Extend-C18

    TABLE 1Characteristics of ZORBAX Rx-SIL and a Contrasting Type of Silica

    FIGURE 1Formation of ZORBAX Porous Silica Particles

  • 3

    CH2-OH

    CH2-OH

    CH2CH3

    CH2CH3

    NH2

    NH2

    The Benefit of Silica Purity Reduced Peak Tailing

    Peak tailing of basic compounds canbe a major chromatographic problem.Peak tailing reduces chromatographicefficiency and the accuracy and reproducibility of results. The majorcause of peak tailing is interactionsbetween analytes and the silica surface (Figure 2). Typically thepresence of acidic silanol sites on the silica surface cause this type ofpeak tailing. Trace metals in silicaincrease silanol acidity and peakasymmetry. These silanol interactionsare reduced or eliminated by choos-ing a less acidic, ultrapure (99.995%) silica, such as ZORBAX Rx-SIL.

    The improvement in chromatographyis dramatic. Figure 3 shows thereduction in peak tailing using ZORBAX Rx-SIL versus a more acidic silica.

    The Benefits of Strong Particles Greater Efficiency and Durability

    During the silica manufacturingprocess, the agglutinated sol particlesare sintered for increased strength.This improved mechanical stabilityallows ZORBAX columns to be packedat high pressures up to 8000 psi!This results in a packed column withan exceptionally stable column bedthat will not compress under normalor even high operating pressures upto 5000 psi. This packed bed stabilityincreases column lifetime using ZORBAX 3.5, 5 or 7 m particles.When ZORBAX Rapid Resolution, 3.5 m silica particles are used as the underlying support, high speed,high efficiency chromatography is possible without compromising column lifetime.

    The Benefits of Careful Pore Size andParticle Size Control High Efficiencyand Better Reproducibility with MoreColumn Choices

    Accurate and closely monitored parti-cle and pore size control for ZORBAXRx-SIL produces reproducible reten-tion behavior from column-to-columnand lot-to-lot. The narrow, consistentparticle size distribution of ZORBAXRx-SIL particles maximizes efficiencyand column bed stability. Column pres-sure is never unusually high due tofines - smaller particles at the lowend of the particle size distribution.

    Accurate and precise control of particle size allows specific 3.5, 5, and 7 m ZORBAX Rx-SIL particles to be produced. The small 3.5 m particle size is the basis for the Rapid Resolution, high-speed analysis

    SiO-Na+ + R3NH+ SiO- NH+R3 + Na

    +

    -SiOH + RCOO- -SiO- ...H+ ... -OOCR

    1. Ionized silanols (SiO-) will ion-exchange with protonated bases (R3NH

    +) which can cause tailing and method variability.

    2. Unprotonated acids can compete for H+ with protonated silanols.

    SilicaParticle

    SilicaParticle

    SilicaParticle

    SilicaParticle

    columns designed to maximize resolution in shorter column lengths ideal for LC/MS or any applicationdemanding shorter analysis times.The 5 m particles are an industrystandard and provide high resolutionin a wide variety of column dimen-sions. This particle size also provideshigh efficiency in a short preparativeconfiguration the CombiHT column because careful particlesize control means consistent pressure expectations within normaloperating limits. The 7 m particleprovides the ideal balance betweenefficiency and operating pressure for longer preparative columns.

    ZORBAX Rx-SIL The Foundation for Many Bonded-Phases

    With such strong performance characteristics, ZORBAX Rx-SIL particles have been developed intomany effective bonded phases forsolving key analytical problems. These include columns that can beused at extremes of pH, unmatched by any other silica-based columns.Because silica-based columns havedifferent limitations at low and highpH, specific bonded-phase chemistriesare required to provide longer columnlife over different pH ranges.

    As a result, Agilent ZORBAX RP-HPLCbonded phases are designed to giveextended column lifetime and reproducibility in the pH ranges thatprovide optimum and long lasting resolution, all starting with high performance ZORBAX Rx-SIL.

    FIGURE 2Potential Secondary Interactions with Silica Silanols and Ionizable Compounds

    FIGURE 3Chromatographic Improvement UsingHighly Purified ZORBAX Rx-SIL

    Original ZORBAX SIL (1973)

    Highly Purified ZORBAX Rx-SIL

    Mobile Phase: 5% Propanol in Heptane

    Flow Rate: 2.0 mL/min

  • Recommended ColumnZORBAX Extend-C18

    pH Range for Optimal Column Life1 2 3 4 5 6 7 8 9 10 11 12

Recommended

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