The Column Advisor recommends the most appropriate columns depending on the application requirements you need to consider or the column characteristics you are looking for. Click on the appropriate tab and move your mouse on the left hand side table to reveal the corresponding application or column characteristics in the middle of the screen.

• Method Attributes

  Compound retention and compound loading are two important attributes that help decide which column will be most useful for your separation.  The properties and attributes listed here help us recommend a column or column family for you based on the method attributes that you consider being important.  Choosing a column based on retention is analyte specific; the column choice is tailored to the majority of the compounds of interest.  However, some methods require columns that are tolerant to high mass loading conditions.

• Compound Retention

  Chromatographic sorbents are chosen for their ability to retain a certain class or mixture of compounds.  In most cases, using a column that increases retention will allow you to increase the separation between closely eluting chromatographic peaks.  The columns in this section can be chosen by how well they can retain compound classes as well as those that are suited for polar compound retention.

• Polar Analytes (RP)

  Polar compounds often elute in the void when organic modifiers are used in the mobile phase making accurate quantification difficult or impossible. It is common to use 100% aqueous mobile phases to retain polar compounds, such as metabolites or small organic acids.  The columns listed here are designed to maximize retention under these aqueous mobile phase conditions.
  134529162,134529189,
  • XSelect HSS T3 The best option for transferring methods from HPLC to ACQUITY UPLC. The XSelect HSS T3 columns are available in both HPLC and ACQUITY UPLC particle sizes.
  • Atlantis T3 The best option for reversed-phase HPLC methods and general purpose chromatography.
• Polar Analytes (HILIC)

  HILIC (Hydrophilic Interaction Chromatography) is a technique that is commonly used when reversed-phase columns, such as the Atlantis T3, do not provide enough compound retention, even with 100% aqueous mobile phases.

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  • XBridge Amide The extended pH range of this column is ideally suited for general purpose retention of polar acidic, polar basic or mixtures of compounds. Optimized for separating simple sugars (e.g.: sucrose, glucose) and polysaccharides.
  • Atlantis HILIC This silica based sorbent is the first choice for retaining small polar bases. It is best suited for low pH mobile phases. Available in HPLC particle sizes only.
  • CORTECS HILIC This high efficiency solid-core silica column is designed for retention of extremely polar, basic, water-soluble analytes. It is usable under low pH conditions.
• General Mixtures

  Columns are recommended based on their ability to provide the widest retention capability for polar and non-polar compound mixtures using standard reversed-phase mobile phases.

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  • XSelect HSS T3 The best option for when you desire to transfer methods from HPLC to UPLC in the future. The XSelect HSS T3 columns are available in both HPLC and ACQUITY UPLC particle sizes for easy method transfer between LC platforms.
  • CORTECS C18 The best general purpose option when high-efficiency separations are desired. This solid-core column offers a balanced retention of acidic, basic, and neutral analytes, while providing enhanced resolution for complex mixtures.
  • Atlantis T3 The best option for HPLC methods development and general purpose chromatography. Available in HPLC particle sizes only.
• Non-Polar Basic Analytes

  My recommendation is to use a general purpose column that is capable of using a wide mobile phase pH range, which gives the user the greatest ability to influence compound retention by modifying additive choice.  For example, the general rule for a basic analyte:
  - Low pH mobile phases decrease the retention of bases
  - High pH mobile phases increase the retention of bases

  134528929,134529153,134828118,
  • XBridge BEH C18 This column will give you the maximum retention for basic compounds when using high pH mobile phase buffers. Available in both HPLC and ACQUITY UPLC particle sizes for easy method transfer between LC platforms.
  • XSelect CSH C18 This column is preferred when the majority of your methods are developed for low pH mobile phases. Out of these selections, this column will provide the best peak shape for bases at low pH. Available in both HPLC and ACQUITY UPLC particle sizes.
  • CORTECS C18+ Use this column for high-efficiency separations at low pH. The charged surface of the particle enables excellent peak shape of basic compounds at low pH, especially in the presence of low ionic strength modified mobile phases.
• Non-Polar Acidic Analytes

  Retaining non-polar acidic compounds is most achievable using specialized phases other than C18; however, reproducibility and column lifetimes have been the two biggest concerns for users.  Generally, these columns are used with low pH mobile phases.

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  • XSelect CSH Fluoro-Phenyl This unique fluorinated ligand provides the best retention for highly acidic compounds (i.e. strong acids) at low pH.
  • XBridge BEH Shield RP18 Generally less retentive for charged species compared to the XSelect Fluoro-Phenyl column, this column does well for weak acids and is more retentive that a standard alkyl C18 column
  • XBridge BEH C18 Even though this is the least retentive option, this column provides the longest lifetime at low pH. It is for this reason we recommend it as the best option for routine LC/MS applications.
• Compound Loading

  Compound loading ability is an indicator of how well a chromatographic sorbent can tolerate extreme mass loading of an analyte before resolution or peak shape is compromised.  High load capabilities are important in areas such as impurity profiling and natural product isolation and are valued by preparative and dissolution scientists.

• Acidic Analytes

  For acidic analytes, the best chromatographic performance under high loading conditions is achieved using low pH mobile phases and columns that are designed for chemical stability under these conditions.

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  • XSelect CSH C18 Provides alternative selectivity to the SunFire C18 column. This new hybrid-based column also is suitable for high load performance and has been optimized for low pH separations. Available in both HPLC and ACQUITY UPLC particle sizes.
  • SunFire C18 This silica-based column provides superior loading and lifetime under low pH conditions. Available in HPLC and preparative formats only.
• Basic Compounds

  For basic analytes, there are two ways to maintain separation and peak shape under high loading conditions.  For the highest analyte loading for basic compounds, many users prefer to use high pH mobile phases.  The high pH mobile ensures that the analyte is in a neutral charge state, thus maximizing reversed-phase retention and reducing any unwanted ionic interactions with the sorbent substrate. When the compound is chemically unstable at high pH, it is common to use ion-pairing agents and low mobile phase pH conditions. 

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  • XSelect CSH C18 Often selected for low pH applications without the need for unnecessary mobile phase additives, such as TFA. Available in both HPLC and ACQUITY UPLC particle sizes for easy method transfer between LC platforms.
  • XBridge BEH C18 Selected for high pH applications because the highest mass loading for bases is often obtained when the compound is eluted in its neutral state at high pH. Available in both HPLC and ACQUITY UPLC particle sizes.
  • SunFire C18 A silica-based option that provides excellent loading and alternative selecivity for bases at low pH. Available in HPLC and preparative formats only.

• Column Attributes

  In some circumstances you will need a column to perform based on its chemical properties to improve analyte-stationary phase interactions. The column properties listed here are the most commonly referred to characteristics that aid chromatographic separations.

• Selectivity

  Selectivity refers to the extent to which a column can separate compounds in mixtures without interferences from other components.  In general, most methods are developed using a classical C18 phase.  More recent phases, such as those containing embedded polar groups, provide alternative selectivity, while providing similar retention.

• Classic C18 Behavior

  Columns that exhibit C18-like selectivity are classified using the USP designation L1, and are often referred to as “ODS” phases.  These general purpose columns are widely used to solve the most common problems for reversed-phase separations.

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  • CORTECS C18 The best general purpose option when high-efficiency separations are desired. This solid-core column offers a balanced retention of acidic, basic, and neutral analytes, while providing enhanced resolution for complex mixtures.
  • XBridge BEH C18 The best choice for applications that exclusively use high pH mobile phases. An excellent high pH choice for high column lifetimes.
  • XSelect HSS C18 This classic silica-based option that provides alternative selectivity for users who desire a general purpose C18 phase suitable for low pH applications.
• Specialty C18

  These columns are classified with a USP L1 designation; however, they contain specialized ligand bonding or endcapping that gives preferential selectivity to a particular compound class. These columns include embedded polar (shield) columns and columns designed for polar retention. 

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  • XBridge BEH Shield RP18 Contains a polar embedded group within the C18 ligand that provides alternative selectivity and sharper peaks compared to an alkyl C18, such as XBridge C18 (e.g., increased retention for phenolic compounds).
  • XSelect HSS C18 SB This column contains a standard silica-based alkyl C18 ligand that is non-endcapped exposing active silanol sites, which increases interactions for basic analytes.
  • XSelect HSS T3 The premier column for reversed-phase polar compound retention.
• Alternate to C18

  Columns that contain alternative ligands have markedly different selectivity when compared to standard reversed-phase C18 columns.  They are ideally suited for method development, where it is desirable to discriminate between critical pairs or to significantly increase the separation of chromatographic interferences.

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  • XSelect HSS PFP A silica-based perfluorinated phenyl column designed to maximize selectivity and retention for polar and non-polar basic compounds. Available in both HPLC and ACQUITY UPLC particle sizes for easy method transfer between LC platforms.
  • XSelect HSS CN This column chemistry can be used for multiple modes of chromatographic separation, including reversed-phase, normal-phase and HILIC separations. Available in both HPLC and ACQUITY UPLC particle sizes for easy method transfer between LC platforms.
  • XSelect CSH Fluoro-Phenyl Designed to maximize selectivity differences, for acidic compounds. Available in both HPLC and ACQUITY UPLC particle sizes for easy method transfer between LC platforms. (USP L43)
  • XSelect CSH Phenyl-Hexyl XSelect CSH Phenyl-Hexyl columns (USP L11) provide you with alternative analyte selectivity for polyaromatic compounds through pi-pi interactions. Available in both HPLC and ACQUITY UPLC particle sizes for easy method transferbetween LC platforms.
  • XBridge Phenyl These columns provide complementary selectivity to C18 stationary phases, especially for polyaromatic compounds. Available in both HPLC and ACQUITY UPLC particle sizes for easy method transfer between LC platforms. (USP L11)
• Non Endcapped Columns

  A non-endcapped stationary phase can provide different analyte selectivity compared to one that is endcapped, although sometimes at the expense of chromatographic peak shape and pH stability.  A non-endcapped stationary phase exposes un-bonded silanol groups for potential interaction with the analyte.  Such chemistries are valuable method development tools.

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  • XSelect HSS C18 SB This column contains a standard silica-based alkyl C18 ligand that is non-endcapped exposing active silanol sites. This column is categorized as a USP L1 column.
  • XSelect HSS PFP This silica-based column contains perfluorophenyl ligand that is non-endcapped. This column is categorized as a USP L43 column.
  • XSelect CSH Fluoro-Phenyl This column is an alternative to the traditional C18 column and is classified as USP L43 column. The non-encapped fluorinated phase provides alternative selectivity to standard LC separations.
  • XSelect HSS CN This column is an alternative to the traditional C18 column and is classified as USP L10 column. The non-encapped cyano phase provides alternative selectivity to standard LC separations.
• Stability

  The stability of a chromatographic sorbent refers to its ability to resist chemical attack from the mobile phase under a variety of operating conditions.  Chemical attack, including ligand loss through hydrolysis and particle dissolution, results in reduced column lifetime, peak deterioration, and changes in analyte selectivity.

• Low pH

  Column lifetime performance using mobile phases with pH less than 3 can be controlled by optimizing ligand density and bonding.  Under these acidic conditions a tri-functionally bonded ligand and chemically stable base particle will outperform a mono- or di- functionally bonded stationary phase.

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  • XSelect HSS C18 This column is selected when you require a silica-based reversed-phase column option for alternative selectivity.
  • XBridge BEH C18 Is the most modern option available for general purpose LC separations at low pH. The nature of the hybrid-based particle used in this column has reduced silanol activity compared to a similar silica-based C18 column.
  • CORTECS C18 The best general purpose option when high-efficiency separations are desired. This solid-core column offers a balanced retention of acidic, basic, and neutral analytes, while providing enhanced resolution for complex mixtures.
• High pH

  Column lifetime performance is often compromised when using mobile phases that exceed pH 7 and most silica based columns begin to degrade rapidly beyond pH 8.  Hybrid based particles with high ligand densities exhibit the longest lifetimes and provide the ability to routinely use mobile phases that exceed pH 10 for extended periods.

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  • XBridge C18 The most modern option available for general purpose LC separations using high pH mobile phases.
  • XBridge BEH Phenyl This column is selected when you require an alternative to XBridge C18. The highly stable phenyl ligand provides complimentary selectivity especially for polyaromatic compounds.
• Specific Requirement...

  Some columns are designed to have characteristics that can be applied to common application needs, such as general purpose LC/MS columns or those specific to biomolecular analysis.

• LC/MS Recommended

  Most modern LC columns are compatible with MS instrumentation; however, column bleed can compromise MS performance.  It is for this reason that following columns are selected for their mechanical and chemical stability under a wide range of operating conditions.

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  • XBridge BEH C18 Provides the widest usable pH range and the best column lifetimes for general purpose LC/MS work. These columns are ideally suited for volatile buffers between pH 2 and 11.
  • CORTECS C18+ Use this column for high-efficiency separations at low pH. The charged surface of the particle enables excellent peak shape of basic compounds at low pH, especially in the presence of low ionic strength modified mobile phases.
  • XSelect HSS T3 These columns are selected when you require a silica-based column option for alternative selectivity. These columns are designed to be used for both polar and non polar acid, neutral and basic compounds.
• Efficiency

  Column efficiency or plate count measures peak dispersion. A highly efficient column will generate chromatograms with tall, sharp peaks and enhance resolution, speed and sensitivity. The following high performance columns utitilize solid-core technology, which provides an efficiency advantage when compared to similar columns of a fully porous nature.

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  • CORTECS C18+ Use this column for high-efficiency separations at low pH. The charged surface of the particle enables excellent peak shape of basic compounds at low pH, especially in the presence of low ionic strength modified mobile phases.
  • CORTECS C18 The best general purpose option when high-efficiency separations are desired. This solid-core column offers a balanced retention of acidic, basic, and neutral analytes, while providing enhanced resolution for complex mixtures.
  • CORTECS HILIC This high efficiency solid-core silica column is designed for retention of extremely polar, basic, water-soluble analytes. It is usable under low pH conditions.
• Best Peakshape for Bases

  The best performing and most desirable columns maintain good peak shape for basic compounds under a wide set of operating conditions, including the use of reduced ionic-strength mobile phases (e.g: 0.1% formic acid).  These columns will also maintain this symmetrical peak shape as the analyte mass load in increased.

  134529153,134529172,134828118,
  • XSelect CSH C18 These columns are specifically designed for use with non-buffered, low ionic strength mobile phases. The controlled positive charge on the CSH particle reduces the effect of high mass load and is the best choice for when you need to scale the separation
  • CORTECS C18+ Use this column for high-efficiency separations at low pH. The charged surface of the particle enables excellent peak shape of basic compounds at low pH, especially in the presence of low ionic strength modified mobile phases.
  • SunFire C18 These columns are selected when you require a silica-based column. The proprietary bonding and endcapping used for SunFire columns provides symmetrical peak shapes and improved resolution for acidic, neutral and basic compounds at low to intermediate pH
• Biomolecule and Wide Pore

  These columns are design and QC tested for biomolecular separations, specifically protein and peptides using reversed-phase conditions.  These columns have wider pore volumes making them more suitable for these larger molecule separations.

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  • XBridge Peptide BEH300 C18 The XBridge BEH300 C18 Peptide Separation Technology (PST) columns are optimized and QC tested for peptide separations.
  • XBridge Protein BEH300 C4 Waters Protein Separation Technology (PrST) columns are QC tested and optimized to separate proteins based on size, hydrophobicity and isoelectric point.
  • XBridge Peptide BEH130 C18 The XBridge BEH130 C18 Peptide Separation Technology (PST) columns are optimized and QC tested for peptide separations.
  • XBridge Oligonucleotide BEH C18 Waters Oligonucleotide Separation Technology (OST) columns are used to separate detritylated synthetic oligonucleotides based on ion-pair, reversed-phase chromatography.
• HILIC

  HILIC (Hydrophilic Interaction Chromatography) is a secondary technique that is commonly used when specialized reversed-phase polar retentions columns, such as the Atlantis T3, do not provide enough compound retention, even with 100% aqueous mobile phases.
  Polar stationary phases, such as unbonded silica are typically used with polar solvents such as acetonitrile to achieve separation of polar analytes.  Stronger retention is observed when the polarity of the stationary increases (hybrid particle versus silica particle) and the polarity of the mobile phase decreases (methanol versus acetonitrile).

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  • Atlantis HILIC This silica based sorbent is the first choice for retaining small polar bases; however, it is limited to low pH mobile phases. Available in HPLC formats only.
  • CORTECS HILIC This high efficiency solid-core silica column is designed for retention of extremely polar, basic, water-soluble analytes. It is usable under low pH conditions.
  • XBridge BEH Amide The first choice for sugar and carbohydrate analysis. It is also recommended for retaining polar acids and neutral polar compound mixtures. Available in HPLC and UPLC formats.
  • XBridge BEH HILIC This hybrid based sorbent has less HILIC retention than the Atlantis HILIC. It is used when you require more pH stability under more aggressive mobile phases conditions (pH range: 1-9). Available in HPLC and UPLC formats for easy method transfer.