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ACE® CN-ES

Mechanism of Separation	Strength of Interaction
Dipole-dipole interactions	Strong
Hydrophobic binding interactions	Strong

Target Analytes
·Polar and non-polar analytes
·Analytes with double and triple bonds
·Analytes differing in hydrophobicity
·Polar molecules having insufficient retention on traditional CN bonded phases

Recommended Separations
Mixtures of very polar, polar and non-polar analytes
Suitable for NP and RP separations
Separations where traditional CN bonded phases show insufficient stability/lifetime
Applications where a typical C18 column does not provide adequate separation
As an orthogonal phase in method development

Fig. 1 - ACE CN-ES Provides Alternative Selectivity Application # 1701

Sample: 1) metronidazole 2) benzyl alcohol 3) hydrochlorothiazide 4) vanillin
5) methyl paraben 6) 1,2-dinitrobenzene
Mobile Phase: A = 0.1% formic acid in H2O B = 0.1% formic acid in 90:10 MeOH/H2O
Gradient: 3 - 100% B in 5 minutes
Column Dimensions: 50 x 2.1mm Flow Rate: 0.60ml/min
Temperature: 40°C Wavelength: 254nm

Why does ACE CN-ES provide alternative selectivity?

■ Combines a polar CN group with an extended spacer that provides retention similar to a C18 column
■ Extended spacer technology additionally provides increased column lifetime compared to traditional CN bonded phases with short alkyl spacer (typically C3/propyl)

ACE CN-ES provides retention similar to a C18 column

Fig. 2 - Separation of Neutral Compounds under RP conditions Application # 1702

Sample: 1) uracil 2) dimethyl phthalate 3) toluene 4) biphenyl 5) phenanthrene Mobile Phase: 60:40 MeCN/H2O
Column Dimensions: 150 x 4.6mm Flow Rate: 1.00ml/min Temperature: 22°C Wavelength: 254nm

Alternative Selectivity with MeOH or MeCN

Fig. 3 - Advantages of Multi-Mode Interactions on Separations Application # 1703

Sample: 1) 1,3-dihydroxybenzene 2) catechol 3) hydrochlorothiazide 4) oxyprenolol 5) salicylic acid 6) myricetin 7) piroxicam 8) 1,2-dinitrobenzene 9) tolmetin 10) 1-naphthol
11) piperine 12) diflunisal 13) propylbenzene 14) 1,2,3-trichlorobenzene Mobile Phase: A = 0.1% formic acid in H2O B = 0.1% formic acid in MeCN Gradient: 3 - 100% B in 10 minutes
Column Dimensions: 100 x 2.1mm Flow Rate: 0.60ml/min Temperature: 40°C Wavelength: 210nm

Fig. 4 - Fast Gradient with NSAIDs Application # 1704

Exploit Enhanced Selectivity for UHPLC

Fig. 5 - Steroids by UHPLC Application # 1705

Sample: 1) cortisone 2) corticosterone 3) 11α-hydroxyprogesterone 4) cortisone-21-acetate 5) 11-ketoprogesterone 6) β-estradiol 7) 17α-estradiol 8) 17α-ethynylestradiol 9) estrone
Mobile Phase: A = 0.1% formic acid in H2O B = 0.1% formic acid in MeCN Gradient: 25 – 52.5% B in 5 minutes
Column Dimensions: 50 x 2.1mm Flow Rate: 0.40ml/min Temperature: 40°C Wavelength: 260nm

ACE CN-ES is additionally compatible with Normal Phase conditions

Fig. 6 - Retention Comparison with Substituted Aromatic Compounds under NP Conditions Application # 1706

Product Availability and Specifications

ACE CN-ES Capillary and Nano HPLC Columns
ACE CN-ES capillary and nano columns are available in 1mm, 0.5mm (500μm), 300μm, 100μm and 75μm internal diameters with 3μm, 5μm and 10μm particle sizes.
Please enquire for further information.

UHPLC and HPLC Method Development Kits

Method development kits enable the optimum bonded phase for an application to be identified. ACE columns are available with a unique range of highly selective phases in
2μm, 3μm, 5μm and 10μm particle sizes, specially developed for challenging UHPLC and HPLC applications.
The availability of a range of phases offering complementary selectivity enables resolution to be maximised, improves the chances of impurity detection and increases the speed at which methods can be systematically developed. Please enquire for further information.

UHPLC and HPLC Column Accessories
UHPLC Column Connectors

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-CC10, 10 pack) enable the inlet end of UHPLC columns to be correctly installed every time. Their unique reuseable design
ensures that they maintain a 1700 bar (25000 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 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-CC10, 10 pack, see overleaf) may alternatively be used.

UHPLC Pre-column Filters
To further extend column lifetime under UHPLC conditions up to 15000 psi (1000 bar), ACE Excel UHPLC Pre-column filters are recommended. By one estimate, over
70% of the failures of UHPLC and HPLC columns are caused by inlet frit plugging.Due to their ultra low dispersion design, column performance and retention remain
unaffected, whilst the column inlet is protected from blockage. ACE Excel UHPLC Pre-column filters can be installed on any brand of UHPLC column in seconds.
Simply finger tighten initially, then wrench tighten a further ? turn. Two UHPLC pre-column filter options are available, for compatibility with all UHPLC systems:
■ ACE Excel UHPLC Pre-column Filters (p/n EXL-PCF10, 10 pack) are compatible with almost all UHPLC system configurations.
■ For inlet connections onto a Waters Acquity system (containing a Waters Acquity 1/16” fitting and ferrule on the inlet tubing) the use of a pre-column filter incorporating the Waters Acquity column port profile is alternatively recommended (p/n EXL-PCF10/ACQ - 10 pack) to ensure maximum compatibility.

■ 1000 bar (15000 psi) pressure rating
■ Eliminates poor connections
■ Compatible with all UHPLC column brands
■ Compatible with all UHPLC systems including Waters Acquity

HPLC Column Connectors

ACE Fingertight HPLC Column Connectors (p/n ACE-CC10, 10 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 350 bar/5000 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.

■ Fingertight to 350 bar (5000 psi)
■ Reuseable and simple to install
■ Eliminates poor connections
■ Compatible with all

HPLC Guard Cartridges

Guard cartridges are recommended to prevent both inlet frit blockage and irreversible sample adsorption onto the top of the column. Guard cartridges are available for all
ACE HPLC columns from nano and capillary dimensions through to preparative scale.
■ Pressure rated to 275 bar (4000 psi)
■ Simple to install and exchange
■ Increase HPLC column lifetime

HPLC Pre-column Filters

As an alternative to guard cartridges, pre-column filters may be used to protect the column inlet frit from blockage. Due to their ultra low dispersion design, column
performance and retention remain unaffected. ACE HPLC pre-column filters utilise a PEEK? fingertight design that connects directly into any 1/16” 10-32 internal thread column inlet, and are thus compatible with all HPLC column brands.
For 3.0 - 4.6mm id HPLC columns, the use of ACE Analytical HPLC Pre-column Filters (p/n ACE-CS210, 10 pack) are recommended. For 2.1mm id HPLC columns, the use of
ACE Microbore HPLC Pre-column Filters (p/n ACE-HP210, 10 pack) are recommended.
ACE HPLC pre-column filters are simply hand tightened into the column inlet to achieve a pressure rating of 350 bar (5000 psi). If higher pressure rating is required,
ACE Excel UHPLC Pre-column Filters (suitable for use up to 1000 bar/15000 psi) are alternatively recommended.
■ Pressure rated to 350 bar (5000 psi)
■ Simple to install and replace
■ Effective low cost column protection
■ Compatible with all HPLC column brands

ACE® and Excel® are registered trademarks of Advanced Chromatography Technologies Limited. Comparative data may not be representative of all applications.
Advanced Chromatography Technologies Limited acknowledges the registered and unregistered trademarks of Victrex Plc and Waters Corporation and has no association with
either company.