ACE. Intelligent Solutions for Method Development. Method Development Kits. ACE UHPLC and HPLC Columns. a a

Transcription

1 r: ff e ts ice O ki r ti y mn e p tiv olu sam mn c c u le the col e d S an for ngle le si ab s a l i a a av ACE Method Development Kits Intelligent Solutions for Method Development Highly cost effective UHPLC and HPLC method development kits Kits available from microbore to analytical dimensions Porous, solid-core, biomolecule and HILIC options Excellent peak shape, efficiency, reproducibility and lifetime Wide range of particle sizes and complementary phases available Now Available NEW HILIC Method Development Kits ACE UHPLC and HPLC Columns

2 ACE Method Development Kits Intelligent Solutions for Method Development Highly cost effective - ACE Method Development Kits are available for the same price as a single column! different ACE Method Development Kits available from microbore (.mm id) through to analytical (.mm id) dimensions for rapid, systematic method development. Each kit contains carefully selected ACE phases which enables the power of selectivity to be fully exploited. Each ACE phase provides different selectivity due to differing interactions. Bonded Phase Hydrophobic Binding Separation Mechanism and Relative Strength π-π Interaction Dipole- Dipole Hydrogen Bonding Shape Selectivity ACE Advanced Method Development Kit (see pages -) ACE Extended Method Development Kit (see pages -) ACE C **** - - * ** ACE C-AR **** *** (donor) * ** *** ACE C-PFP **** *** (acceptor) **** *** **** ACE SuperC **** ** ACE C-Amide **** - ** **** **/*** ACE CN-ES *** * *** ** * ACE UltraCore Method Development Kit (see pages -) ACE UltraCore SuperC ACE UltraCore SuperPhenylHexyl *** ** ** *** (donor) * ** *** ACE Bioanalytical ACE C- ** - - * * Å Method Development Kit ACE C- * (see pages -) ACE Phenyl- * ** (donor) * ** ** Approximate value determined by semi-quantitative mechanism weightings and/or by reference to other ACE phases using > characterising analytes. Separation Mechanism and Relative Strength Bonded Phase Partitioning Anionic Analyte Interactions Cationic Analyte Interactions Attraction Repulsion Attraction Repulsion H-bonding ACE HILIC Method Development Kit (see pages -) ACE HILIC-A ** - *** **** - * ACE HILIC-B *** **** - - *** * ACE HILIC-N **** **** Approximate value determined by semi-quantitative mechanism weightings and/or by reference to other ACE phases using > characterising analytes. FREE Method Development Support! Not sure which ACE phase or kit will work best for your application? FREE Application Support and FREE Method Development Service Trust your method development to our experts and free up time for your other projects! Contact our expert method development team via info@ace-hplc.com or contact your local distributor Learn More: Columns

3 Selectivity Offer: and column kits available for the same price as a single column Why Use ACE Method Development Kits? ACE HPLC/UHPLC columns have earned a well deserved reputation for delivering excellent efficiency, reproducibility and lifetime. ACE Method Development Kits group together columns with different mechanisms of interaction to maximise selectivity and improve the likelihood of separating difficult or closely related analytes in mixtures. Screening columns containing different bonded phases under the same mobile phase conditions can help you achieve your desired separation more quickly, therefore increasing productivity. Highly cost effective to promote bonded phase investigation as part of an intelligent method development strategy, ACE Method Development Kits are available for the same price as a single column! Si-OH ACE Stationary Phases Virtually Eliminate the Negative Effects of Silanols on UHPLC & HPLC Separations Using ACE Method Development Kits to Improve Separations Columns Within ACE Method Development Kits Provide Alternative Selectivity Application #, ACE C,, ACE C provides excellent peak shape, but here the essentially hydrophobic-only interaction results in co-elution. Investigation of alternative bonded phases using the same test conditions is recommended. Time - ACE C-PFP ACE C-PFP provides additional interactions compared to alkyl C phases. Whilst these change selectivity, in this instance co-elution of different analyte pairs is observed. Time - ACE C-AR The ACE C-AR phase provides a further change in selectivity due to different interaction contributions ultimately enabling a successful separation. Screening alternative phases can maximise selectivity and reduce method development time. Time - Sample: ) metronidazole ) -hydroxybenzoic acid ) -hydroxybenzoic acid ) benzyl alcohol ) benzoic acid ) myrecetin ) p-cresol ) propranolol ) ethyl paraben ) furosemide ) anisole ),,-trinitrobenzene ) toluene ) nimesulide ) mefenamic acid ),,-trichlorobenzene Mobile Phase: A =.% formic acid in H O B =.% formic acid in MeCN Gradient: % B in minutes Column Dimensions: x.mm Flow Rate:.ml/min Temperature: C Detection: nm

4 ACE Advanced Method Development Kit Contains ACE C, ACE C-AR and ACE C-PFP phases Ideal starting point for routine method development Available from microbore (.mm id) through to analytical (.mm id) dimensions (see p.) Particularly recommended for compounds containing aromatic rings Phase Functional Group Endcapped Particle Size (µm) Pore Size (Å) Surface Area (m /g) Carbon Load (%) Recommended ph Range % Aqueous Compatible USP Listing ACE C Octadecyl (C) Yes.,,,,..-. a No L ACE C-AR ACE C-PFP C with integral Phenyl C with integral PFP Yes.,,,,..-. a Yes L Yes.,,,,..-. a Yes L a For optimum column lifetime, a ph range of - is recommended. To increase column lifetime at higher ph, organic buffers, low buffer concentrations, high % organic solvent and low temperatures must be considered. Further information is contained within A Guide to HPLC and LC/MS Buffer Selection by John Dolan please contact your distributor to request your FREE copy or visit. ACE C ACE C-AR ACE C-PFP ACE C remains the go-to column of choice for HPLC and UHPLC separations. With an excellent reputation for performance, reproducibility and lifetime, ACE C provides a rugged, reproducible starting point for method development. Recommended Applications Analytes differing in hydrophobicity Polar, moderately polar and non-polar analytes Uncharged acids and bases Ionized acids or bases using ion-pairing Ideal starting point for method development ACE C-AR combines the excellent performance and advantages of the ACE C phase with the added selectivity of an integral phenyl group. Recommended Applications Analytes with π-bonding and conjugated systems Analytes with electron delocalization and electron withdrawing groups, such as halogens, nitro groups, ketones, esters and acids Analytes with different dipole moments Analytes differing in hydrophobicity Stereoisomers, steroids, substituted aromatics and sulphur containing compounds Fully wettable - % aqueous buffer compatible Applications where C does not provide adequate separation Applications where conventional phenyl phases provide insufficient retention, poor stability, or significant bleed. ACE C-PFP brings together the stability, reproducibility and low bleed of the ACE C phase with the additional selectivity of an integral pentafluorophenyl (PFP) group. Recommended Applications Analytes with π-bonding Analytes with electron donating groups, such as phenols, aromatic ethers and amines Analytes with proton donor groups Analytes with different dipole moments Analytes differing in hydrophobicity Stuctural isomers, steroids, substituted aromatics and taxanes Fully wettable - % aqueous buffer compatible Applications where C does not provide adequate separation Applications where conventional PFP phases provide insufficient retention, poor stability or significant bleed. Additional Information Product bulletins containing further details on the ACE C, C-AR and C-PFP columns contained within the Advanced ACE Method Development Kit are available to download at Alternatively, please contact our technical support team via info@ace-hplc.com or contact your local distributor. Learn More: ACE HPLC and UHPLC Columns Product Catalogue ACE

5 ACE C - Comparison of Column Inertness Column brands from major manufacturers investigated Comparison of column efficiency for pyridine a basic molecule Peak Efficiency Comparison Application # ACE C Sunfire. C HyPurity C Zorbax XDB. C Zorbax SB. C Luna C() XBridge. C Inertsil ODS- XTerra MS. C Ascentis Express. C,,,,,,,,,, Hypersil BDS C Spherisorb ODS,, Symmetry. C, Hypersil ODS,,,,, N. (pyridine) (plates/m) Si-OH Column Dimensions: x.mm Sample: ) uracil ) pyridine ) phenol Mobile Phase: : (v/v) MeOH/H O Flow Rate:.ml/min Temperature: C Detection: nm Comparative data may not be representative of all applications. Please see back page for acknowledgement of trademarks. ACE Stationary Phases Virtually Eliminate the Negative Effects of Silanols on UHPLC & HPLC Separations ACE C Delivers Excellent Performance Rapid UHPLC Screening of Pharmaceuticals and Related Compounds Application # ACE Excel C (fully porous ultra-inert silica) P max = bar ZORBAX Eclipse. XDB C (fully porous silica), P max = bar These leading C phases provide an essentially hydrophobic-only interaction, and therefore exhibit similar selectivity with only slight differences between brands observed. Waters ACQUITY. BEH C (hybrid particle) Phenomenex Kinetex. C (core shell particle),, P max = bar P max = bar To implement a change in selectivity, the use of an alternative bonded phase (eg ACE C-AR, ACE C-PFP) that can leverage additional modes of interaction is recommended. Sample: ) N-acetylprocainamide ) -hydroxybenzoic acid ) pindolol ) methylphenylsulphoxide ) benzyl alcohol ) quinoxaline ),-dinitrobenzene ) phenacetin ),-dimethoxybenzene ) furosemide ) anisole ) methyl benzoate ) remacemide ) nimesulide ) ethyl benzoate ) diflunisal Mobile Phase: A = mm KH PO, ph. B = mm KH PO, ph. in MeOH/H O (: v/v) Gradient: % B in minutes Column Dimensions: x.mm Flow Rate:.ml/min Temperature: C Detection: nm Comparative data may not be representative of all applications. Please see back page for acknowledgement of trademarks.

6 Leveraging the Unique Selectivity of ACE C-AR Improving an Analgesics Separation by Changing Phase Application #,, ACE C,, C phase provides essentially hydrophobic-only interaction ACE C-AR Multi-mode interaction including π - π and hydrophobic interactions provides complete separation Sample: ) -acetamidophenol ) -aminobenzoic acid ) -hydroxybenzoic acid ) caffeine ) -acetamidophenol ) -hydroxybenzoic acid ) salicylamide ) acetanilide ) phenol ) acetylsalicylic acid ) benzoic acid ) sorbic acid ) salicylic acid ) phenylacetin ) salicylaldehyde Mobile Phase: A =.% v/v formic acid in H O B =.% v/v formic acid in MeCN Gradient: - % B in minutes, hold at % B until minutes Column Dimensions: x.mm Flow Rate:.ml/min Temperature: C Detection: nm ACE C-PFP Provides a Separation that a C or PFP Column Alone Cannot Achieve The Importance of Maintaining Hydrophobicity During Multi-Mode Interactions Application # ACE C-PFP Multi-mode interaction including hydrophobic interaction provides complete separation, ACE C, C phase provides essentially hydrophobic-only interaction Hypersil GOLD µm PFP (example short chain PFP phase) Reduced Hydrophobicity Hydrophobic reference Significantly reduced hydrophobicity reduces the separation despite other interaction modes Column Dimensions: x.mm Sample: ),,-trimethoxybenzene ),,-trimethoxybenzene ),-dimethoxybenzene ),-dimethoxybenzene ) methoxybenzene ),-dimethoxybenzene ),,-trimethoxybenzene ) neutral molecule (reference) Mobile Phase: : v/v MeOH/H O Flow Rate:.ml/min Temperature: C Detection: nm Comparative data may not be representative of all applications. Please see back page for acknowledgement of trademarks.

7 ACE Advanced Method Development UHPLC/HPLC Column Kits (Contains columns: ACE C, ACE C-AR and ACE C-PFP of specified dimensions) (UHPLC/HPLC hardware format with bar/psi pressure limit) Column Dimensions.µm µm µm µm. x mm MDKA--U MDKA--U MDKA--U MDKA--U. x mm MDKA--U MDKA--U MDKA--U MDKA--U. x mm MDKA--U MDKA--U MDKA--U MDKA--U. x mm MDKA--U MDKA--U MDKA--U MDKA--U. x mm MDKA--U MDKA--U MDKA--U MDKA--U. x mm MDKA--U MDKA--U MDKA--U MDKA--U. x mm - MDKA--U MDKA--U MDKA--U. x mm - MDKA--U MDKA--U MDKA--U. x mm - - MDKA--U MDKA--U. x mm MDKA--U MDKA--U MDKA--U MDKA--U. x mm MDKA--U MDKA--U MDKA--U MDKA--U. x mm MDKA--U MDKA--U MDKA--U MDKA--U Selectivity Offer:. x mm MDKA--U MDKA--U MDKA--U MDKA--U. x mm MDKA--U MDKA--U MDKA--U MDKA--U. x mm - MDKA--U MDKA--U MDKA--U Each column kit is available for the same price as a single column. x mm MDKA--U MDKA--U MDKA--U MDKA--U. x mm - MDKA--U MDKA--U MDKA--U. x mm - - MDKA--U MDKA--U. x mm - MDKA--U MDKA--U MDKA--U. x mm - MDKA--U MDKA--U MDKA--U. x mm - MDKA--U MDKA--U MDKA--U. x mm - MDKA--U MDKA--U MDKA--U. x mm - MDKA--U MDKA--U MDKA--U. x mm - MDKA--U MDKA--U MDKA--U. x mm - MDKA--U MDKA--U MDKA--U. x mm - MDKA--U MDKA--U MDKA--U. x mm - - MDKA--U MDKA--U ACE Advanced Method Development Microbore HPLC Column Kits (Contains columns: ACE C, ACE C-AR and ACE C-PFP of specified dimensions) (HPLC hardware format with bar/psi recommended pressure limit) µm µm µm Column Dimensions / port / port / port / port / port / port. x mm MDKA-- MDKA--S MDKA-- MDKA--S MDKA-- MDKA--S. x mm MDKA-- MDKA--S MDKA-- MDKA--S MDKA-- MDKA--S. x mm MDKA-- MDKA--S MDKA-- MDKA--S MDKA-- MDKA--S. x mm MDKA-- MDKA--S MDKA-- MDKA--S MDKA-- MDKA--S. x mm MDKA-- MDKA--S MDKA-- MDKA--S MDKA-- MDKA--S. x mm MDKA-- MDKA--S MDKA-- MDKA--S MDKA-- MDKA--S. x mm MDKA-- MDKA--S. x mm MDKA-- MDKA--S MDKA-- MDKA--S MDKA-- MDKA--S. x mm MDKA-- MDKA--S MDKA-- MDKA--S MDKA-- MDKA--S. x mm MDKA-- MDKA--S MDKA-- MDKA--S MDKA-- MDKA--S. x mm MDKA-- MDKA--S MDKA-- MDKA--S MDKA-- MDKA--S. x mm MDKA-- MDKA--S MDKA-- MDKA--S MDKA-- MDKA--S. x mm MDKA-- MDKA--S MDKA-- MDKA--S MDKA-- MDKA--S. x mm MDKA-- MDKA--S Important Note: ACE microbore columns (.mm id and.mm id) are available with either standard / (- thread) connections or / (- thread) connections. For use with Eksigent micro and nano LC systems, order columns with / connections and use either ACE - fittings (part number ACE-MC, pack) or Eksigent - fittings (part number ). For / HPLC column connections up to psi, PEEK / fingertight fittings (part number ACE-CC, pack) are recommended. For / microbore HPLC column connections up to psi, PEEK / (- thread) fingertight fittings (part number ACE-MC, pack) are recommended. For / UHPLC column connections up to psi, reuseable / fittings (part number EXL-CC, pack) are recommended. To further extend UHPLC and HPLC column lifetimes, ACE pre-column filters are recommended. For further details please contact your distributor or visit

8 Exploit selectivity at low, intermediate and high ph Use with LC/MS compatible buffers between ph.. High efficiency µm, µm, µm and µm particles for UHPLC and HPLC Ultra-inert for maximum performance and reproducibility UHPLC and HPLC Columns Alternate selectivity for method development Improved separations with polar, acidic, basic and phenolic compounds High efficiency µm, µm, µm and µm particles for UHPLC and HPLC Ultra-inert for maximum performance and reproducibility ACE C-Amide for increased polar retention and alternate selectivity UHPLC and HPLC Columns ACE Extended Method Development Kit Contains ACE SuperC, ACE C-Amide and ACE CN-ES phases Use ACE SuperC to exploit selectivity changes at low, intermediate and high ph Available from microbore (.mm id) through to analytical (.mm id) dimensions (see p.) ACE C-Amide and ACE CN-ES phases both offer alternative selectivity, especially for polar molecules Phase Functional Group Endcapped ACE SuperC Octadecyl (C) Encapsulated bonding Particle Size (µm) Pore Size (Å) Surface Area (m /g) Carbon Load (%) Recommended ph Range % Aqueous Compatible USP Listing.,,,,..-. a No L ACE C-Amide ACE CN-ES C with integral amide polar group CN with proprietary extended alkyl spacer Yes.,,,,..-. b Yes L/L Yes.,,,,..-. b Yes L a ACE SuperC is designed for use with LC/MS compatible buffers. Further information is contained within ACE SuperC - A Guide to Buffer Selection please contact your distributor to request your FREE copy or visit. b For optimum column lifetime, a ph range of - is recommended. To increase column lifetime at higher ph, organic buffers, low buffer concentrations, high % organic solvent and low temperatures must be considered. Further information is contained within A Guide to HPLC and LC/MS Buffer Selection by John Dolan please contact your distributor to request your FREE copy or visit ACE SuperC ACE C-Amide ACE CN-ES ACE SuperC is a uniquely bonded, EBT endcapped C phase which offers unprecedented inertness, excellent efficiency and uncompromising durability over an extended ph range of... ACE C-Amide is a uniquely designed polar-embedded phase that offers enhanced retention and resolution of polar acidic, phenolic and hydroxysubstituted analytes. The extended spacer ligand technology provides extended column lifetime. ACE CN-ES is a unique phase having an extended alkyl chain with a terminal cyano group. It provides C levels of retention and stability compared to commercial cyano propyl phases which typically exhibit low retentivity and poor stability. Recommended Applications Analytes differing in hydrophobicity Polar, moderately polar and non-polar analytes Uncharged acids and bases Ionized acids or bases using ion-pairing Recommended starting point for developing methods at intermediate and high ph to exploit selectivity changes Recommended Applications Small water soluble analytes and polar molecules - especially acidic species Analytes with H bond donors, acids, bases and phenolic compounds Small peptides Analytes differing in hydrophobicity Fully wettable - % aqueous buffer compatible Applications where C does not provide adequate separation Applications where conventional amide/ polar embedded phases provide insufficient retention, poor stability, or significant bleed Recommended Applications Mixtures of very polar, polar and nonpolar analytes Analytes with double and triple bonds Analytes differing in hydrophobicity Suitable for NP and RP separations Fully wettable - % aqueous buffer compatible Applications where a typical C column does not provide adequate separation Applications where traditional CN bonded phases provide insufficient retention, poor stability or significant bleed An orthogonal phase for method development Additional information Product bulletins containing further details on the ACE SuperC, C-Amide and CN-ES columns contained within the Extended ACE Method Development Kit are available to download at Alternatively, please contact our technical support team via info@ace-hplc.com or contact your local distributor. Learn More: TM ACE SuperC Ultra-Inert UHPLC and HPLC Columns with Extended ph Stability ACE ACE C-Amide For increased polar retention and alternate selectivity

9 ACE SuperC Provides Excellent Column Inertness Leading column brands in x.mm LC/MS compatible dimensions at intermediate ph. Silica, Hybrid and Superficially Porous particle technologies compared Comparison of column efficiency for pyridine a basic molecule Efficiency measured at % peak height to account for peak tailing effects Peak Efficiency Comparison Application # Reproduced with kind permission of The Open University, UK ACE Excel SuperC,pl/m ZORBAX Eclipse. XDB C Agilent Poroshell. EC C,pl/m,pl/m Higher Efficiency Waters XBridge. BEH C XP,pl/m Waters ACQUITY. BEH C,pl/m Phenomenex Kinetex. C,pl/m Waters XBridge. C,pl/m Phenomenex Luna. C() HST a,pl/m NUCLEOSHELL. RP a,pl/m Phenomenex Kinetex. C ZORBAX Extend. C,pl/m,pl/m Lower Efficiency Thermo Accucore. C,pl/m,,,,, N. (pyridine) (plates/m) Column Dimensions: x.mm ( a x.mm) Sample: ) uracil ) pyridine ) phenol Mobile Phase: : v/v MeOH/mM NH OAc in H O (ph.) Flow Rate:.ml/min Temperature: C Detection: nm Comparative data may not be representative of all applications. Please see back page for acknowledgement of trademarks. Use ACE SuperC to Investigate ph Effects Exploit Selectivity by Adjusting ph Application # ph ph ph. ph ph ph ph ph ph. ph ph ph ph Column: ACE Excel µm SuperC, x.mm Sample: ) nizatidine ) salbutamol ) amiloride ) N-acetylprocainamide ) quinoxaline ) methyl paraben ) p-cresol ) reserpine ) piperine ) toluene ) felodipine Temperature: C Flow Rate:.ml/min Wavelength: nm Gradient: - % B in minutes Acidic Mobile Phase: A: mm ammonium formate in H O (ph.) B: mm ammonium formate (ph.) in : v/v MeCN/H O Basic Mobile Phase: A:.% NH (= mm) in H O (ph.) B:.% NH (=mm), ph. in : v/v MeCN/H O ph

10 ACE C-Amide Provides Enhanced Polar Selectivity Advantages of Multi-Mode Interactions for HPLC Separations Application # Reproduced with kind permission of The Open University, UK ACE C-Amide (ultra-inert fully porous silica) ACE C-Amide provides increased retention and improved separation due to enhanced polar selectivity ACE C (ultra-inert fully porous silica), Waters ACQUITY. BEH C (hybrid particle) Leading C column brands provide similar selectivity, Phenomenex Kinetex. C (superficially porous particle) Column Dimensions: x.mm Sample: ) resorcinol ) catechol ) -methyl resorcinol ) -methyl catechol ) -methyl catechol ) -nitro catechol Mobile Phase: : MeOH/mM H PO in H O Flow Rate:.ml/min Temperature: C Wavelength: nm Comparative data may not be representative of all applications. Please see back page for acknowledgement of trademarks. ACE CN-ES Provides Alternative Selectivity ACE CN-ES Provides a Separation that a C or CN Column Alone Cannot Achieve Application # ACE CN-ES ACE CN-ES phase provides complete separation due to hydrophobic AND dipole-dipole interactions,, ACE CN (example short chain CN phase) CN phase shows insufficient resolution (greatly reduced hydrophobic interaction) ACE C, C phase provides insufficient resolution (no dipole-dipole interaction) Sample: ) metronidazole ) benzyl alcohol ) hydrochlorothiazide ) vanillin ) methyl paraben ),-dinitrobenzene Mobile Phase: A =.% formic acid in H O B =.% formic acid in : MeOH/H O Gradient: - % B in minutes Column Dimensions: x.mm Flow Rate:.ml/min Temperature: C Wavelength: nm

11 ACE Extended Method Development UHPLC/HPLC Column Kits (Contains columns: ACE SuperC, ACE C-Amide and ACE CN-ES of specified dimensions) (UHPLC/HPLC hardware format with bar/psi pressure limit) Column Dimensions.µm µm µm µm. x mm MDKE--U MDKE--U MDKE--U MDKE--U. x mm MDKE--U MDKE--U MDKE--U MDKE--U. x mm MDKE--U MDKE--U MDKE--U MDKE--U. x mm MDKE--U MDKE--U MDKE--U MDKE--U. x mm MDKE--U MDKE--U MDKE--U MDKE--U. x mm MDKE--U MDKE--U MDKE--U MDKE--U. x mm - MDKE--U MDKE--U MDKE--U. x mm - MDKE--U MDKE--U MDKE--U. x mm - - MDKE--U MDKE--U. x mm MDKE--U MDKE--U MDKE--U MDKE--U. x mm MDKE--U MDKE--U MDKE--U MDKE--U. x mm MDKE--U MDKE--U MDKE--U MDKE--U. x mm MDKE--U MDKE--U MDKE--U MDKE--U. x mm MDKE--U MDKE--U MDKE--U MDKE--U. x mm MDKE--U MDKE--U MDKE--U MDKE--U. x mm - MDKE--U MDKE--U MDKE--U. x mm - MDKE--U MDKE--U MDKE--U. x mm - - MDKE--U MDKE--U. x mm - MDKE--U MDKE--U MDKE--U. x mm - MDKE--U MDKE--U MDKE--U. x mm - MDKE--U MDKE--U MDKE--U. x mm - MDKE--U MDKE--U MDKE--U. x mm - MDKE--U MDKE--U MDKE--U. x mm - MDKE--U MDKE--U MDKE--U. x mm - MDKE--U MDKE--U MDKE--U. x mm - MDKE--U MDKE--U MDKE--U. x mm - - MDKE--U MDKE--U Selectivity Offer: Each column kit is available for the same price as a single column ACE Extended Method Development Microbore HPLC Column Kits (Contains columns: ACE SuperC, ACE C-Amide and ACE CN-ES of specified dimensions) (HPLC hardware format with bar/psi recommended pressure limit) µm µm µm Column Dimensions / port / port / port / port / port / port. x mm MDKE-- MDKE--S MDKE-- MDKE--S MDKE-- MDKE--S. x mm MDKE-- MDKE--S MDKE-- MDKE--S MDKE-- MDKE--S. x mm MDKE-- MDKE--S MDKE-- MDKE--S MDKE-- MDKE--S. x mm MDKE-- MDKE--S MDKE-- MDKE--S MDKE-- MDKE--S. x mm MDKE-- MDKE--S MDKE-- MDKE--S MDKE-- MDKE--S. x mm MDKE-- MDKE--S MDKE-- MDKE--S MDKE-- MDKE--S. x mm MDKE-- MDKE--S. x mm MDKE-- MDKE--S MDKE-- MDKE--S MDKE-- MDKE--S. x mm MDKE-- MDKE--S MDKE-- MDKE--S MDKE-- MDKE--S. x mm MDKE-- MDKE--S MDKE-- MDKE--S MDKE-- MDKE--S. x mm MDKE-- MDKE--S MDKE-- MDKE--S MDKE-- MDKE--S. x mm MDKE-- MDKE--S MDKE-- MDKE--S MDKE-- MDKE--S. x mm MDKE-- MDKE--S MDKE-- MDKE--S MDKE-- MDKE--S. x mm MDKE-- MDKE--S IMPORTANT NOTE: ACE microbore columns (.mm id and.mm id) are available with either standard / (- thread) connections or / (- thread) connections. For use with Eksigent micro and nano LC systems, order columns with / connections and use either ACE - fittings (part number ACE-MC, pack) or Eksigent - fittings (part number ). For / HPLC column connections up to psi, PEEK / fingertight fittings (part number ACE-CC, pack) are recommended. For / microbore HPLC column connections up to psi, PEEK / (- thread) fingertight fittings (part number ACE-MC, pack) are recommended. For / UHPLC column connections up to psi, reuseable / fittings (part number EXL-CC, pack) are recommended. To further extend UHPLC and HPLC column lifetimes, ACE pre-column filters are recommended. For further details please contact your distributor or visit

12 ACE UltraCore Method Development Kits Contains ACE UltraCore SuperC and SuperPhenylHexyl phases Use to exploit selectivity changes at low, intermediate and high ph Available from microbore (.mm id) through to analytical (.mm id) dimensions (see p.) Ultra inert core-shell particles and Encapsulated Bonding Technology (EBT ) provide excellent peak shape Phase Functional Group Particle Size (µm) Pore Size (Å) Surface Area (m /g) Carbon Load (%) Maximum ph Range ACE UltraCore. SuperC Octadecyl encapsulated...-. a L ACE UltraCore. SuperPhenylHexyl Phenyl-Hexyl encapsulated...-. a L ACE UltraCore SuperC Octadecyl encapsulated..-. a L ACE UltraCore SuperPhenylHexyl Phenyl-Hexyl encapsulated..-. a L USP Listing a ACE UltraCore columns are designed for use with LC/MS compatible buffers. Further information is contained within ACE UltraCore A Guide to Buffer Selection - please contact your distributor to request your FREE copy or visit. ACE UltraCore - Columns Provide Improved Peak Shape ACE UltraCore SuperC and SuperPhenylHexyl phases are manufactured using our unique Encapsulated Bonding Technology (EBT ) This technology dramatically increases ligand coverage of the silica surface and effectively eliminates the negative effects of unbonded silanol groups The higher ligand coverage results in improved inertness, chromatographic performance and stability Use ACE UltraCore to Investigate ph Effects Si-OH ACE UltraCore Stationary Phases Virtually Eliminate the Negative Effects of Silanols on UHPLC & HPLC Separations Exploit Selectivity by Adjusting ph Application #, ph ph ph. ph ph ph ph ph ph. ph ph ph ph Column: ACE UltraCore.µm SuperC, x.mm Sample: ) atenolol ) methylphenylsulphoxide ) eserine ) prilocaine ) bupivacaine ) tetracaine ),,,-tetrahydro--naphthol ) carvedilol ) nitrobenzene ) methdilazine ) amitriptyline ) valerophenone Temperature: C Flow Rate:.ml/min Wavelength: nm Gradient: % B in minutes Acidic Mobile Phase: A: mm ammonium formate in H O (ph.) B: mm ammonium formate (ph.) in : (v/v) MeCN/H O Basic Mobile Phase: A:.% NH (= mm) in H O (ph.) B:.% NH (=mm), ph. in : (v/v) MeCN/H O ph

13 ACE UltraCore Columns are Highly Inert Solid-core columns from leading manufacturers investigated Comparison of column efficiency for pyridine a basic molecule Peak Efficiency Comparison Application # Reproduced with kind permission of The Open University, UK Pressure Efficiency ACE UltraCore. SuperPhenylHexyl bar,pl/m Higher Efficiency ACE UltraCore. SuperC Phenomenex Kinetex. C Phenomenex Kinetex. C Waters Cortecs. C Phenomenex Kinetex. C bar bar bar bar bar,pl/m,pl/m,pl/m,pl/m,pl/m Thermo Accucore. C bar,pl/m,,,,, N. (pyridine) (plates/m) Lower Efficiency Column Dimensions: x.mm Sample: ) uracil ) pyridine ) phenol Mobile Phase: : v/v MeOH/mM NH OAc in H (ph.) Flow Rate:.ml/min Temperature: C Detection: nm Comparative data may not be representative of all applications. Please see back page for acknowledgement of trademarks. Introducing Selectivity Changes Using ACE UltraCore Method Development Kits Vitamin Separation Application #, ACE UltraCore SuperC ACE UltraCore SuperPhenylHexyl Multi-mode interaction (including π-π and hydrophobic interactions) provides complete separation..... Column: x.mm,.µm Sample: ) pyridoxine ) p-aminobenzoic acid ) pantothenic acid ) folic acid ) d-biotin ) cyanocobalamin ) riboflavin Mobile Phase A: mm KH PO ph. (aq) B: mm KH PO ph. in MeOH:H O : (v/v) Gradient: Time (mins).... %B Flow Rate:. ml/min Temperature: C Wavelength: nm min

14 ACE UltraCore Method Development UHPLC/HPLC Column Kits (Contains columns: ACE UltraCore SuperC and ACE UltraCore SuperPhenylHexyl of specified dimensions) (UHPLC/HPLC hardware format with bar/psi pressure limit) Column Dimensions.µm µm. x mm MDKU--U MDKU--U. x mm MDKU--U MDKU--U. x mm MDKU--U MDKU--U. x mm MDKU--U MDKU--U. x mm MDKU--U MDKU--U. x mm MDKU--U MDKU--U. x mm MDKU--U MDKU--U. x mm MDKU--U MDKU--U. x mm - MDKU--U. x mm MDKU--U MDKU--U. x mm MDKU--U MDKU--U. x mm MDKU--U MDKU--U. x mm MDKU--U MDKU--U. x mm MDKU--U MDKU--U. x mm MDKU--U MDKU--U. x mm MDKU--U MDKU--U. x mm MDKU--U MDKU--U. x mm - MDKU--U. x mm MDKU--U MDKU--U. x mm MDKU--U MDKU--U. x mm MDKU--U MDKU--U. x mm MDKU--U MDKU--U. x mm MDKU--U MDKU--U. x mm MDKU--U MDKU--U. x mm MDKU--U MDKU--U. x mm MDKU--U MDKU--U. x mm - MDKU--U Selectivity Offer: Each column kit is available for the same price as a single column ACE UltraCore Method Development Microbore HPLC Column Kits (Contains columns: ACE UltraCore SuperC and ACE UltraCore SuperPhenylHexyl of specified dimensions) Column Dimensions (HPLC hardware format with bar/psi recommended pressure limit).µm µm / port / port / port / port. x mm MDKU-- MDKU--S MDKU-- MDKU--S. x mm MDKU-- MDKU--S MDKU-- MDKU--S. x mm MDKU-- MDKU--S MDKU-- MDKU--S. x mm MDKU-- MDKU--S MDKU-- MDKU--S. x mm MDKU-- MDKU--S MDKU-- MDKU--S. x mm MDKU-- MDKU--S MDKU-- MDKU--S. x mm - - MDKU-- MDKU--S. x mm MDKU-- MDKU--S MDKU-- MDKU--S. x mm MDKU-- MDKU--S MDKU-- MDKU--S. x mm MDKU-- MDKU--S MDKU-- MDKU--S. x mm MDKU-- MDKU--S MDKU-- MDKU--S. x mm MDKU-- MDKU--S MDKU-- MDKU--S. x mm MDKU-- MDKU--S MDKU-- MDKU--S. x mm - - MDKU-- MDKU--S IMPORTANT NOTE: ACE microbore columns (.mm id and.mm id) are available with either standard / (- thread) connections or / (- thread) connections. For use with Eksigent micro and nano LC systems, order columns with / connections and use either ACE - fittings (part number ACE-MC, pack) or Eksigent - fittings (part number ). For / HPLC column connections up to psi, PEEK / fingertight fittings (part number ACE-CC, pack) are recommended. For / microbore HPLC column connections up to psi, PEEK / (- thread) fingertight fittings (part number ACE-MC, pack) are recommended. For / UHPLC column connections up to psi, reuseable / fittings (part number EXL-CC, pack) are recommended. To further extend UHPLC and HPLC column lifetimes, ACE pre-column filters are recommended. For further details please contact your distributor or visit

15 ACE Bioanalytical Å Method Development Kits Contain ACE C-, ACE C- and ACE Phenyl- phases Ideal starting point for protein and peptide method development Available from microbore (.mm id) through to analytical (.mm id) dimensions (see p. ) Ultra-inert Å phases provide excellent peak shape and reproducibility Phase Functional Group Particle Size (µm) Pore Size (Å) Surface Area (m /g) Carbon Load (%) Recommended ph Range USP Listing ACE C- Octadecyl (C),,..-. a L ACE C- Butyl (C),,..-. a L ACE Phenyl- Phenyl,,..-. a L a For optimum column lifetime, a ph range of - is recommended. To increase column lifetime at higher ph, organic buffers, low buffer concentrations, high % organic solvent and low temperatures must be considered. Further information is contained within A Guide to HPLC and LC/MS Buffer Selection by John Dolan please contact your distributor to request your FREE copy or visit. ACE Å Ultra-Inert Columns Provide Improved Peak Shape ACE Å ultra-inert HPLC columns are manufactured using advanced technology that virtually eliminates the negative effects of silanols and metal contamination for the separation of peptides, proteins and other high molecular weight biomolecules. The ultra-inert characteristics of ACE Å columns permit the use of as little as.% TFA in the mobile phase. Lower purity columns show unacceptable peak tailing even when using as much as.% TFA. The ability to run at reduced TFA concentrations results in increased sensitivity. ACE Å Bioanalytical Columns Provide Excellent Peak Shape Application # ACE C- (Ultra-inert silica) Å, C Column (Lower purity active silica).% TFA.% TFA.% TFA Columns: x.mm, µm Sample: ) Gly-Tyr ) Oxytocin ) Angiotensin II ) Neurotensin Mobile Phase: A = TFA in H O (% as specified above) B = TFA in MeCN (% as specified above) Gradient: - % B in. minutes Flow Rate:.ml/min Temperature: C Wavelength: nm Comparative data may not be representative of all applications. Please see back page for acknowledgement of trademarks.

16 Using ACE Bioanalytical Å Method Development Kits to Optimise Selectivity Introducing Selectivity Changes by Careful Consideration of Bonded Phase and Mobile Phase Additive.% TFA.% Formic Acid Application # ACE C- Response - Millivolts Response - Millivolts,, ACE C- ACE Phenyl- Response - Millivolts Response - Millivolts Response - Millivolts Response - Millivolts The ACE Å C, C and Phenyl chemistries contained within ACE Bioanalytical Å Method Development Kits enable the analyst to investigate selectivity effects due to phase variations. The ultra-inert characteristics of the ACE Å silica enable different mobile phase additives to be investigated without a deterioration in peak shape or sensitivity. Columns: x.mm, µm Sample: ) Gly-Tyr ) Val-Tyr-Val ) Methionine enkephalin ) Angiotensin II ) Leucine enkephalin Mobile Phase: A =.% TFA in H O or.% Formic Acid in H O (as specified above) B = MeCN Gradient: % B in minutes Flow Rate:.ml/min Temperature: C Wavelength: nm

17 ACE Bioanalytical Å Method Development HPLC Column Kits (Contains columns: ACE C-, ACE C- and ACE Phenyl- of specified dimensions) (HPLC hardware format with bar/psi pressure limit) Column Dimensions µm µm. x mm MDKB-- MDKB--. x mm MDKB-- MDKB--. x mm MDKB-- MDKB--. x mm MDKB-- MDKB--. x mm MDKB-- MDKB--. x mm MDKB-- MDKB--. x mm MDKB-- MDKB--. x mm MDKB-- MDKB--. x mm - MDKB--. x mm MDKB-- MDKB--. x mm MDKB-- MDKB--. x mm MDKB-- MDKB--. x mm MDKB-- MDKB--. x mm MDKB-- MDKB--. x mm MDKB-- MDKB--. x mm MDKB-- MDKB--. x mm MDKB-- MDKB--. x mm - MDKB--. x mm MDKB-- MDKB--. x mm MDKB-- MDKB--. x mm MDKB-- MDKB--. x mm MDKB-- MDKB--. x mm MDKB-- MDKB--. x mm MDKB-- MDKB--. x mm MDKB-- MDKB--. x mm MDKB-- MDKB--. x mm - MDKB-- Selectivity Offer: Each column kit is available for the same price as a single column Note:.mm id ACE Bioanalytical Å Method Development Kits also available please enquire ACE Bioanalytical Å Method Development Microbore HPLC Column Kits (Contains columns: ACE C-, ACE C- and ACE Phenyl- of specified dimensions) (HPLC hardware format with bar/psi recommended pressure limit) µm µm Column Dimensions / port / port / port / port. x mm MDKB-- MDKB--S MDKB-- MDKB--S. x mm MDKB-- MDKB--S MDKB-- MDKB--S. x mm MDKB-- MDKB--S MDKB-- MDKB--S. x mm MDKB-- MDKB--S MDKB-- MDKB--S. x mm MDKB-- MDKB--S MDKB-- MDKB--S. x mm MDKB-- MDKB--S MDKB-- MDKB--S. x mm - - MDKB-- MDKB--S. x mm MDKB-- MDKB--S MDKB-- MDKB--S. x mm MDKB-- MDKB--S MDKB-- MDKB--S. x mm MDKB-- MDKB--S MDKB-- MDKB--S. x mm MDKB-- MDKB--S MDKB-- MDKB--S. x mm MDKB-- MDKB--S MDKB-- MDKB--S. x mm MDKB-- MDKB--S MDKB-- MDKB--S. x mm - - MDKB-- MDKB--S IMPORTANT NOTE: ACE microbore columns (.mm id and.mm id) are available with either standard / (- thread) connections or / (- thread) connections. For use with Eksigent micro and nano LC systems, order columns with / connections and use either ACE - fittings (part number ACE-MC, pack) or Eksigent - fittings (part number ). For / HPLC column connections up to psi, PEEK / fingertight fittings (part number ACE-CC, pack) are recommended. For / microbore HPLC column connections up to psi, PEEK / (- thread) fingertight fittings (part number ACE-MC, pack) are recommended. For / UHPLC column connections up to psi, reuseable / fittings (part number EXL-CC, pack) are recommended. To further extend UHPLC and HPLC column lifetimes, ACE pre-column filters are recommended. For further details please contact your distributor or visit

18 ACE HILIC Method Development Kits Contains ACE HILIC-A, ACE HILIC-B and ACE HILIC-N phases Alternative and improved selectivity to reversed-phase for polar and very polar analytes Available from microbore (.mm id) through to analytical (.mm id) dimensions (see p. ) ACE HILIC-A, ACE HILIC-B and ACE HILIC-N provide alternative selectivity to each other Phase Functional Group Endcapped Particle Size (µm) Pore Size (Å) Surface Area (m /g) Carbon Load (%) Recommended ph Range ACE HILIC-A Proprietary SIL No.,, -.-. L ACE HILIC-B ACE HILIC-N Proprietary Aminopropyl Proprietary Polyhydroxy USP Listing No.,,..-. L No.,,..-. Pending To extend column lifetime under HPLC conditions (up to psi / bar), ACE guard cartridges (/pk) or ACE pre-column filters (/pk) are recommended. For the guard system a guard cartridge holder (H) and coupler (C) are also required. To extend column lifetime under UHPLC conditions (up to psi / bar) an ACE UHPLC pre-column filter is recommended. What is HILIC? Hydrophilic Interaction Liquid Chromatography (HILIC) was first described by Alpert*. HILIC is ideal for the separation and retention of polar species including polar neutral and polar ionised analytes. HILIC separations typically include a polar stationary phase with high organic solvent containing mobile phases. Mechanistically HILIC is complex (Fig ) and provides multiple modes of interaction between the analyte, stationary phase, eluent and water enriched layer at the stationary phase particle-eluent interface**. * A. J. Alpert, J. Chromatogr., (). ** See the FREE ACE guide to reproducible HILIC method development for more information order your copy now. Charged Analyte Electrostatic Attraction Polar Neutral Analyte Neutral Analyte Partitioning H-bonding Stationary Phase Particle Electrostatic Repulsion Water enriched layer Figure. Schematic of interactions between different types of polar analytes and a stationary phase in HILIC mode When Should You Consider HILIC? HILIC provides the retention and separation of hydrophilic or polar to very polar analytes not well retained in RPLC. Hydrophilic or polar to very polar analytes have log P values (measure of lipophilicity) of around zero or less (Fig A). Generally, polar analytes are suitable for HILIC if they elute before caffeine in gradient RPLC (Fig B). Charged Analyte Figure A. Analyte suitability for HILIC from Log P HILIC RPLC << - - Log P Scale >> Figure reproduced with permission and adapted from work first published in Chromatography Today, Volume, Issue, November/December Glyphosate log P = -. Caffeine log P = -. Amitriptyline log P =. mv Glyphosate Caffeine Amitriptyline min Figure B. Analyte suitability for HILIC from gradient RPLC Column: ACE Excel C, x.mm, µm Part Number: EXL --U Mobile Phase: A = mm ammonium formate, ph. (aq) B = mm ammonium formate, ph. in : v/v MeCN:H O Gradient: -% B in minutes Detection: ELSD Flow Rate:.mL min Temperature: ºC Injection: µl Analysed using VWR-Hitachi Chromaster with VWR ELSD.

19 ACE HILIC Columns Alternative Selectivities ACE HILIC-A ACE HILIC-B ACE HILIC-N An acidic character phase with an ionisable negative surface charge depending on mobile phase ph A basic character phase with an ionisable positive surface charge depending on mobile phase ph A neutral character phase capable of H-bonding amongst other mechanisms of interaction ACE HILIC-A An acidic character phase. b-blockers (ACE AN#) mau. Propranolol. Sotalol min.. min. Acebutalol. Salbutamol. Atenolol Column: ACE HILIC-A, x.mm, µm Part Number: HILA--U Mobile Phase: mm ammonium formate ph. in MeCN/H O (: v/v) Flow Rate:.mL/min Injection: µl Temperature: ºC Detection: nm ACE HILIC-B A basic character phase. Succinic acid and methylmalonic acid (ACE AN#) mv Solvent front. Succinic acid. Methylmalonic acid min Column: ACE HILIC-B, x.mm, µm Part Number: HILB--U Mobile Phase: mm ammonium formate ph. in MeCN/H O (: v/v) Flow Rate:.mL/min Temperature: ºC Injection: µl Detection: ELSD (Evaporator temp: ºC, Nebuliser temp: ºC, Gas speed: SLM) ACE HILIC-N A polar neutral character phase. Nucleobases and nucleosides (ACE AN#) mau.e+.e+.e+.e+.e+.e+ min. Deoxyadenosine. Adenine. Adenosine Column: ACE HILIC-N, x.mm, µm Part Number: HILN--U Mobile Phase: mm ammonium formate ph. in MeCN/H O (: v/v) Flow Rate:.mL/min Temperature: ºC Injection: µl Detection: nm

20 ACE HILIC Method Development ACE HILIC columns provide alternative selectivity to each other. The power of systematic screening of different phase chemistries for HILIC method development is seen below. Maximise your HILIC method development success by following the ACE HILIC method development protocol using three optimised ACE HILIC column chemistries protocol available in the FREE HILIC Method Development guide. Advantages of using ACE HILIC Method Development Kits ACE HILIC-A mau,, Explore Selectivity: Method Development Kits available: column kit for the same price as a single column see page min min mau ACE HILIC-B min min Different Elution Orders and Retention Times Observed ACE HILIC-N mau,, min min. Acebutolol. p-aminobenzoic acid. -Hydroxybenzoic acid. Nicotinamide. -Deoxyguanosine. Adenine. Mandelic acid. Tyramine. Atenolol Columns: All x.mm, µm (Part numbers: ACE HILIC-A: HILA--U, ACE HILIC-B: HILB--U, ACE HILIC-N: HILN--U) Mobile Phase: mm ammonium formate ph. in MeCN/H O (: v/v) Flow Rate:.mL/min Temperature: C Detection: nm Sample: ) p-aminobenzoic acid ) -Hydroxybenzoic acid ) Nicotinamide ) Acebutolol ) Adenine ) Mandelic acid ) Tyramine ) Atenolol ) -Deoxyguanosine Conclusions ACE HILIC columns provide alternative selectivity to each other ideal for HILIC method development. FREE HILIC Method Development Technical Guide A page HILIC Method Development Technical Guide illustrating a tried and tested approach to HILIC method development is available. Request your copy today and learn how to develop reproducible and robust HILIC methods simply and efficiently. Available at Alternatively, please contact our technical support team via info@ace-hplc.com

21 ACE HILIC Method Development UHPLC/HPLC Column Kits (Contains columns: ACE HILIC-A, ACE HILIC-B and ACE HILIC-N of specified dimensions) (UHPLC/HPLC hardware format with bar/psi pressure limit) Column Dimensions.µm µm µm. x mm MDKH--U MDKH--U MDKH--U. x mm MDKH--U MDKH--U MDKH--U. x mm MDKH--U MDKH--U MDKH--U. x mm MDKH--U MDKH--U MDKH--U. x mm MDKH--U MDKH--U MDKH--U. x mm MDKH--U MDKH--U MDKH--U. x mm - MDKH--U MDKH--U. x mm - MDKH--U MDKH--U. x mm - MDKH--U MDKH--U. x mm MDKH--U MDKH--U MDKH--U. x mm MDKH--U MDKH--U MDKH--U. x mm MDKH--U MDKH--U MDKH--U. x mm MDKH--U MDKH--U MDKH--U. x mm MDKH--U MDKH--U MDKH--U. x mm MDKH--U MDKH--U MDKH--U. x mm - MDKH--U MDKH--U. x mm - MDKH--U MDKH--U. x mm - MDKH--U MDKH--U. x mm - MDKH--U MDKH--U. x mm - MDKH--U MDKH--U. x mm - MDKH--U MDKH--U. x mm - MDKH--U MDKH--U. x mm - MDKH--U MDKH--U. x mm - MDKH--U MDKH--U. x mm - MDKH--U MDKH--U. x mm - MDKH--U MDKH--U. x mm - MDKH--U MDKH--U Selectivity Offer: Each column kit is available for the same price as a single column ACE HILIC Method Development Microbore HPLC Column Kits (Contains columns: ACE HILIC-A, ACE HILIC-B and ACE HILIC-N of specified dimensions) (HPLC hardware format with bar/psi recommended pressure limit) µm µm Column Dimensions / port / port / port / port. x mm MDKH-- MDKH--S MDKH-- MDKH--S. x mm MDKH-- MDKH--S MDKH-- MDKH--S. x mm MDKH-- MDKH--S MDKH-- MDKH--S. x mm MDKH-- MDKH--S MDKH-- MDKH--S. x mm MDKH-- MDKH--S MDKH-- MDKH--S. x mm MDKH-- MDKH--S MDKH-- MDKH--S. x mm - - MDKH-- MDKH--S. x mm MDKH-- MDKH--S MDKH-- MDKH--S. x mm MDKH-- MDKH--S MDKH-- MDKH--S. x mm MDKH-- MDKH--S MDKH-- MDKH--S. x mm MDKH-- MDKH--S MDKH-- MDKH--S. x mm MDKH-- MDKH--S MDKH-- MDKH--S. x mm MDKH-- MDKH--S MDKH-- MDKH--S. x mm - - MDKH-- MDKH--S IMPORTANT NOTE: ACE microbore columns (.mm id and.mm id) are available with either standard / (- thread) connections or / (- thread) connections. For use with Eksigent micro and nano LC systems, order columns with / connections and use either ACE - fittings (part number ACE-MC, pack) or Eksigent - fittings (part number ). For / HPLC column connections up to psi, PEEK / fingertight fittings (part number ACE-CC, pack) are recommended. For / microbore HPLC column connections up to psi, PEEK / (- thread) fingertight fittings (part number ACE-MC, pack) are recommended. For / UHPLC column connections up to psi, reuseable / fittings (part number EXL-CC, pack) are recommended. To further extend UHPLC and HPLC column lifetimes, ACE pre-column filters are recommended. For further details please contact your distributor or visit

22 UHPLC, HPLC and Microbore Column Accessories UHPLC Column Connectors Pressure rating > bar (> psi) Compatible with all UHPLC systems Compatible with all UHPLC column brands Eliminates poor connections Innovative reusable design Wrench Flat ¼ (.cm) Proprietary PEEK cone Reuseable column connector with - threads for / od tubing ACE Excel UHPLC Column Connector (p/n EXL-CC, pack) All UHPLC column brands require correct installation in order to realise maximum column efficiency. To avoid connection problems, permanently swaged fittings are not recommended as they do not allow free movement between the tubing, fitting and column inlet on installation. This can result in a poorly connected column that shows unexpected peak tailing due to the introduction of extra column volume (dead volume) to the system. Alternatively, a leak at the inlet fitting connection may be observed. ACE Excel UHPLC Column Connectors (p/n EXL-CC, pack) enable the inlet end of UHPLC columns to be correctly installed every time. Their unique reuseable design ensures that they maintain a bar ( psi) pressure rating with repeated use, yet do not permanently swage onto the inlet tubing. To maximise the lifetime of the fitting, the use of a torque wrench (p/n EXL-TW) is required. At the outlet end of the UHPLC column (where pressure demands are lower but a correct connection remains important), ACE Fingertight HPLC Column Connectors (p/n ACE-CC, pack, see below) may alternatively be used. Note: For inlet connections onto a Waters Acquity system (containing a Waters Acquity / fitting and ferrule on the inlet tubing) the use of a pre-column filter incorporating the unique Waters Acquity column port profile is alternatively recommended (p/n EXL-PCF/ACQ - pack) to ensure maximum compatibility with the Waters Acquity system fittings. HPLC Column Connectors Fingertight to bar ( psi) Reuseable and simple to install Eliminates poor connections Compatible with all HPLC column brands and instruments ACE Fingertight HPLC Column Connectors (p/n ACE-CC, pack) are recommended for the connection of both the inlet and outlet ends of HPLC columns. Manufactured from premium quality PEEK, the fittings simply hand tighten to provide a perfect column connection, and are pressure rated to bar/ psi. ACE Fingertight HPLC Column Connectors may additionally be used at the outlet end of UHPLC columns, where pressure demands are lower but a correct connection remains important. ACE Fingertight HPLC Column Connector (p/n ACE-CC, pack) Microbore HPLC Column Connectors Fingertight to bar ( psi) Reuseable and simple to install Compatible with both capillary (µm od) and / od connection tubing Reuseable connector with - threads for / od tubing ACE Microbore / Column Connectors (p/n ACE-MC, pack) are recommended for the connection of inlet and outlet ends of ACE Microbore HPLC columns with / ports. Each pack comes complete with fittings and capillary sleeves (which enable the fittings to be used with µm od tubing), plus a fingertight tightening adaptor. These fittings are additionally compatible with any other brand of microbore column terminating with / ports and - threads. ACE Microbore / Column Connector (p/n ACE-MC, pack)

23 Your decision has lasting effects. Ultra-Inert Base-Deactivated UHPLC/HPLC Columns For Performance, Selectivity and Guaranteed Reproducibility ACE and Excel are registered trademarks of Advanced Chromatography Technologies Limited. ACE SuperC, ACE UltraCore, EBT and HSC are trademarks of Advanced Chromatography Technologies Limited. Advanced Chromatography Technologies Limited acknowledges the registered and unregistered trademarks of Alltech Associates Inc., Eksigent Technologies, GL Sciences, MACHEREY-NAGEL GmbH & Co KG, Phenomenex Inc., Sigma-Aldrich Co., Thermo Fisher Scientific, Victrex Plc and Waters Corporation and has no affiliation with any of these companies. Columns

24 ACE ACE products are available through our international network of distributors ACE UHPLC and HPLC Columns Advanced Chromatography Technologies Ltd, Berry Street, Aberdeen, AB HF, Scotland Tel : + () Fax: + () info@ace-hplc.com P----