Atlantis T3 and ACQUITY UPLC HSS T3 Columns Brochure
6
■ Superior polar compound retention ■ Wide usable silica pH range ■ Designed for aqueous mobile phases ■ Long column lifetimes ■ Fully Mass Spectrometry (MS) compatible AtLAntiS t3 AnD ACQUitY UPLC HSS t3 CoLUMnS Atlantis ® T3 and ACQUITY UPLC ® HSS T3 columns are the new industry standards for retaining and separating polar compounds via reversed-phase HPLC and UPLC ® technology. Atlantis T3 HPLC columns are universal silica-based, reversed-phase C 18 columns that not only retain and separate highly polar compounds, but also provide superior performance across a wider pH range. ACQUITY UPLC HSS T3 columns maintain all the advantages of Atlantis T3 HPLC columns but are also certified for use in UPLC separations while providing an additional UPLC chemistry choice. The Atlantis T3 columns are designed to provide longer column lifetimes at low pH as well as improved peak shape for amine-containing bases at pH 7. Waters has created a second pressure-tolerant, UPLC-certified particle. This High Strength Silica (HSS) particle is the first and only 100% silica particle designed specifically for 15000 psi/1000 bar applications. ACQUITY UPLC HSS T3 columns utilize the same T3 bonding and provides similar attributes as Atlantis T3 HPLC columns. T3: REDEFINING POLAR COMPOUND RETENTION IN HPLC AND UPLC
Transcript of Atlantis T3 and ACQUITY UPLC HSS T3 Columns Brochure
EasiEr MEt hod t ransfEr
Since Atlantis T3 HPLC and ACQUITY UPLC HSS T3 columns utilize T3 bonding, HPLC methods can be transferred to UPLC methods. Because Atlantis and HSS particles are different substrates, some method modification and/or optimization may be required. A compelling benefit of UPLC technology is the ease and speed at which this method optimization can occur since UPLC technology offers greater resolution in less time.
Moving from XBridge™ HPLC to ACQUITY UPLC BEH columns is nearly seamless, as the hybrid particle substrates differ only in particle size. However, because of T3 bonding it should be straightforward to make simple adjustments to successfully transfer a separation from an Atlantis T3 HPLC column to an ACQUITY UPLC HSS T3 column.
additional choicEs for UPlc sEParations
Chromatographers continue to embrace UPLC technology as the separation technology of the future. Additional UPLC column chemistries that provide complementary selectivities are needed to provide application-specific solutions. When combined with the ultra-resolution of UPLC, these new UPLC column chemistries will enable the development of fast, robust methods in less time. ACQUITY UPLC HSS T3 columns offer another selectivity choice for UPLC separations.
T3 Bonding Enables HPLC to UPLC Transfer
HPLC Separation on Atlantis T3
UPLC Separation on ACQUITY UPLC HSS T3
CompoundRetention Time (min)
USP Resolution
1. Cytosine 2.77
2. 5-Fluorocytosine 3.58 4.56
3. Uracil 3.93 1.71
4. 5-Fluorouracil 4.59 3.41
5. Guanine 5.92 7.76
6. Thymine 7.63 11.01
7. Adenine 8.39 4.70
Similar Selectivities
CompoundRetention Time (min)
USP Resolution
1. Cytosine 0.34
2. 5-Fluorocytosine 0.44 5.02
3. Uracil 0.48 1.50
4. 5-Fluorouracil 0.56 3.17
5. Guanine 0.80 8.16
6. Thymine 1.25 14.65
7. Adenine 1.33 3.50
T3 Bonding Enables HPLC to UPLC™ Transfer
HPLC on Atlantis® T3
UPLC™ on ACQUITY UPLC™ HSS T3
>80% Reduction in Run Time
0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 min
1 2 3 4 5 6 7 8 9 min
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2
3
4
5
6
7
1
2 3
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1 Cytosine2 5-Fluorocytosine3 Uracil4 5-Fluorouracil5 Guanine6 Thymine7 Adenine
T3 Bonding Enables HPLC to UPLC™ Transfer
HPLC on Atlantis® T3
UPLC™ on ACQUITY UPLC™ HSS T3
>80% Reduction in Run Time
0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 min
1 2 3 4 5 6 7 8 9 min
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1 Cytosine2 5-Fluorocytosine3 Uracil4 5-Fluorouracil5 Guanine6 Thymine7 Adenine
T3 bonding allows for the transfer of HPLC methods to UPLC.
Part Number Particle Size Dimension
186003713 3 µm 1.0 x 50 mm
186003730 5 µm 1.0 x 50 mm
186003714 3 µm 1.0 x 150 mm
186003731 5 µm 1.0 x 150 mm
186003715 3 µm 2.1 x 20 mm IS™
186003732 5 µm 2.1 x 20 mm IS
186003716 3 µm 2.1 x 30 mm
186003733 5 µm 2.1 x 30 mm
186003717 3 µm 2.1 x 50 mm
186003734 5 µm 2.1 x 50 mm
186003718 3 µm 2.1 x 100 mm
186003735 5 µm 2.1 x 100 mm
186003719 3 µm 2.1 x 150 mm
186003736 5 µm 2.1 x 150 mm
186003720 3 µm 3.0 x 20 mm IS
186003737 5 µm 3.0 x 20 mm IS
186003721 3 µm 3.0 x 50 mm
186003738 5 µm 3.0 x 50 mm
186003722 3 µm 3.0 x 100 mm
186003739 5 µm 3.0 x 100 mm
186003723 3 µm 3.0 x 150 mm
186003740 5 µm 3.0 x 150 mm
186003741 5 µm 3.0 x 250 mm
186003724 3 µm 4.6 x 20 mm IS
186003742 5 µm 4.6 x 20 mm IS
186003725 3 µm 4.6 x 30 mm
186003743 5 µm 4.6 x 30 mm
186003726 3 µm 4.6 x 50 mm
186003744 5 µm 4.6 x 50 mm
186003727 3 µm 4.6 x 75 mm
186003745 5 µm 4.6 x 75 mm
186003728 3 µm 4.6 x 100 mm
186003746 5 µm 4.6 x 100 mm
186003729 3 µm 4.6 x 150 mm
186003747 5 µm 4.6 x 150 mm
186003748 5 µm 4.6 x 250 mm
Part Number Particle Size Dimension
1860036953 5 µm 10 x 10 mm Guard
1860037063 10 µm 10 x 10 mm Guard
186003691 5 µm 10 x 50 mm
186003692 5 µm 10 x 100 mm
186003693 5 µm 10 x 150 mm
186003704 10 µm 10 x 150 mm
186003694 5 µm 10 x 250 mm
186003705 10 µm 10 x 250 mm
1860036994 5 µm 19 x 10 mm Guard
1860037104 10 µm 19 x 10 mm Guard
186003696 5 µm 19 x 50 mm OBD™
186003707 10 µm 19 x 50 mm OBD
186003697 5 µm 19 x 100 mm OBD
186003698 5 µm 19 x 150 mm OBD
186003708 10 µm 19 x 150 mm OBD
186003709 10 µm 19 x 250 mm OBD
186004026 5 µm 19 x 250 mm OBD
186003700 5 µm 30 x 50 mm OBD
186003701 5 µm 30 x 75 mm OBD
186003702 5 µm 30 x 100 mm OBD
186003703 5 µm 30 x 150 mm OBD
186003711 10 µm 30 x 150 mm OBD
186003712 10 µm 30 x 250 mm OBD
186004080 5 µm 50 x 50 mm OBD
186004081 5 µm 50 x 100 mm OBD
186004082 5 µm 50 x 150 mm OBD
186004083 10 µm 50 x 50 mm OBD
186004084 10 µm 50 x 100 mm OBD
186004085 10 µm 50 x 150 mm OBD
186004086 10 µm 50 x 250 mm OBD
Part Number Particle Size Dimension
186003751 3 µm 4.6 x 150 mm
186003754 5 µm 4.6 x 150 mm
186003755 5 µm 4.6 x 250 mm
Part Number Particle Size Dimension
1860037561 3 µm 2.1 x 10 mm (2/pk)
1860037591 5 µm 2.1 x 10 mm (2/pk)
1860037572 3 µm 3.9 x 20 mm (2/pk)
1860037602 5 µm 3.9 x 20 mm (2/pk)
1860037582 3 µm 4.6 x 20 mm (2/pk)
1860037612 5 µm 4.6 x 20 mm (2/pk)
ordEring inforMat ion
1 - Requires Sentry Guard Holder WAT0979582 - Requires Sentry Guard Holder WAT0469103 - Requires Cartridge Guard Holder 2890007794 - Requires Cartridge Guard Holder 186000709
Compounds 1. Caftaric acid2. Chlorogenic acid3. Cynarin4. Echinacoside5. Cichoric acid
Columns: ACQUITY UPLC HSS 2.1 x 50 mm, 1.8 µm ACQUITY UPLC BEH 2.1 x 50 mm, 1.7 µmMobile Phase A: 0.1% CF3COOH in H2OMobile Phase B: 0.08% CF3COOH in ACNFlow Rate: 0.5 mL/min Gradient: Time Profile Curve (min) %A %B 0.0 92 8 6 0.1 92 8 6 4.45 50 50 7 4.86 10 90 6 5.0 92 8 6 6.0 92 8 6Injection Volume: 1.0 µLSample Diluent: 50:50 H2O: MeOH with 0.05% CF3COOH Sample Conc.: 100 µg/mLTemperature: 40 ˚CDetection: UV @ 330 nmSampling Rate: 40 pts/secTime Constant: 0.1Instrument: Waters ACQUITY UPLC system with ACQUITY UPLC TUV
Minutes
0.00 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00
1 24 53
BEH C18
BEH C8
BEH Shield RP18
BEH Phenyl
1 24 53
1 24 53
1 2 453
1
2 4 53HSS T3
HSS C18143 52
1 2 45
3 HSS C18 SB
The additional UPLC column selectivity choices provide and facilitate rapid development of robust methods.
Reversed-Phase UPLC Chemistry Choices: Caffeic Acid Derivatives in Echinacea
Part Number Particle Size Dimension
186004055 1.8 µm 2.1 x 50 mm
186004056 1.8 µm 2.1 x 100 mm
■ Superior polar compound retention
■ Wide usable silica pH range
■ Designed for aqueous mobile phases
■ Long column lifetimes
■ Fully Mass Spectrometry (MS) compatible
AtLAntiS t3 AnD ACQUitY UPLC HSS t3 CoLUMnS
Atlantis® T3 and ACQUITY UPLC® HSS T3 columns are the new industry standards for retaining and separating polar compounds via reversed-phase HPLC and UPLC® technology. Atlantis T3 HPLC columns are universal silica-based, reversed-phase C18 columns that not only retain and separate highly polar compounds, but also provide superior performance across a wider pH range. ACQUITY UPLC HSS T3 columns maintain all the advantages of Atlantis T3 HPLC columns but are also certified for use in UPLC separations while providing an additional UPLC chemistry choice.
The Atlantis T3 columns are designed to provide longer column lifetimes at low pH as well as improved peak shape for amine-containing bases at pH 7.
Waters has created a second pressure-tolerant, UPLC-certified particle. This High Strength Silica (HSS) particle is the first and only 100% silica particle designed specifically for 15000 psi/1000 bar applications. ACQUITY UPLC HSS T3 columns utilize the same T3 bonding and provides similar attributes as Atlantis T3 HPLC columns.
t3: rEdEfining Polar coMPoUnd rEt Ent ion in hPlc and UPlc
80% Run Time Reduction!!
Atlantis T3 Analytical Columns Atlantis T3 Preparative Columns Atlantis T3 Sentry Guard Cartridges
Atlantis T3 Method Validation Kits*
ACQUITY UPLC HSS T3 Columns
ACQUITY UPLC HSS T3 Method Validation Kits*
* Contains 3 columns, each packed with a different batch of packing material.
Waters corporation 34 Maple Street Milford, MA 01757 U.S.A. T: 1 508 478 2000 F: 1 508 872 1990 www.waters.com
Waters, The Science of What’s Possible, Atlantis, ACQUITY UPLC, UPLC, ACQUITY, ACQUITY Ultra Performance LC, SunFire, XBridge, IS, OBD, Alliance and Sentry are trademarks of Waters Corporation. All trademarks used are acknowledged. All rights reserved.All other trademarks are the property of their respective owners.
©2007 Waters Corporation. Printed/Produced in the U.S.A.November 2007 720001887EN SC-XX
Part Number Particle Size Dimension
186003535 1.8 µm 1.0 x 50 mm
186003536 1.8 µm 1.0 x 100 mm
186003537 1.8 µm 1.0 x 150 mm
186003944 1.8 µm 2.1 x 30 mm
186003538 1.8 µm 2.1 x 50 mm
186003539 1.8 µm 2.1 x 100 mm
186003540 1.8 µm 2.1 x 150 mm
Minutes
Minutes
As chromatographers around the world continue to realize the benefits of UPLC technology in different application areas, Waters continues to develop new UPLC particles and chemistries to meet these ever-growing needs.
ACQUITY UPLC HSS (High Strength Silica) columns contain the first and only 100% silica particle designed, tested and intended for use in applications up to 15000 psi/1000 bar. The ACQUITY UPLC HSS particle is not an HPLC particle. High pore volume HPLC particles do not posses the mechanical stability necessary to withstand the high column packing and operating pressures of UPLC technology. This rugged and unique UPLC- certified silica particle is specifically designed to withstand the demanding operation conditions encountered in UPLC separations.
Waters is the only manufacturer offering two UPLC-certified particles.
acQUit Y UPlc hss t3 colUMns for Polar coMPoUnd rEt Ent ion
ACQUITY UPLC HSS T3 columns utilize T3 bonding in order to retain and separate polar organic compounds. ACQUITY UPLC HSS T3 columns possess the superior polar compound retention, aqueous mobile phase compatibility and ultra-low MS bleed of Atlantis T3 columns.
When compared to ACQUITY UPLC BEH C18 columns, compounds are more strongly retained on ACQUITY UPLC HSS T3 columns.
Features of the ACQUITY UPLC HSS T3 columns include:
■ Superior polar and non-polar compound retention
■ Aqueous mobile phase compatibility
■ Ultra-low MS bleed
■ Additional selectivity choice for UPLC separations.
acQUit Y UPlc hss colUMnsiMP rov Ed ph 7 PErforManc E
At pH 7, poor peak shape for amine-containing bases and shortened column lifetimes are encountered when using intermediate ligand density C18 columns designed for polar compound retention. Poor peak shape is due to secondary interactions with unreacted silanols that remain present after bonding and endcapping. The proprietary T3 end-capping procedure reacts with more of these active silanols thereby dramatically improving peak shape for bases. Shortened column lifetimes are due to the dissolution of the silica particle substrate by the high pH mobile phase. The more effective and efficient T3 bonding helps “protect” the silica substrate from dissolution, thus providing longer column lifetimes.
When compared to other reversed-phase “polar compound retention columns,” Atlantis T3 columns provide superior pH 7 peak shape.
1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0
Thermo Aquasil C18
Agilent Zorbax® SB-Aq
Atlantis dC18
Phenomenex® Aqua
Phenomenex Synergi™ Hydro-RP
Phenomenex Synergi™ Polar-RP
Shiseido Capcell PAK™ C18 AQ®
GL Sciences Inertsil™ ODS-SP
Atlantis T3
Amitriptyline USP Tailing Factor
Did not elute
t3 Bonding
The superior performance of Atlantis T3 HPLC columns and the ACQUITY UPLC HSS T3 columns is a result of Waters new, advanced T3 bonding process. T3 bonding utilizes a trifunctional C18 alkyl phase bonded at a ligand density that promotes polar compound retention and aqueous mobile phase compatibility. The proprietary T3 endcapping process is much more effective than traditional trimethyl silane (TMS) endcapping.
This unique combination of bonding and endcapping provides superior polar compound retention and aqueous compatibility while also enhancing column performance, lifetime, peak shape and stability.
To learn more about T3 bonding and polar compound retention, please refer to Waters white paper entitled:
Topics in Liquid Chromatography Part 1. Designing a Reversed-Phase Column for Polar Compound Retention
Part Number 720001889EN
0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00
1 2 3 45
Atlantis T3
12
4 3,5
Agilent Zorbax® SB-Aq
1 2 4
3
5
Thermo Aquasil C18
12,4
3 5
Phenomenex® Synergi™ Polar - RP
12
Phenomenex® Synergi™ Hydro- RP
3,45
1 23 4 5
Shiseido Capcell Pak™ C18 AQ®
Atlantis T3 columns provide 100% aqueous compatibility, superior polar compound retention and excellent peak shape for acids, neutrals and bases.
Compounds 1. Thiourea 4. Guanosine - 5’- monophosphate2. 5-Fluorocytosine 5. Thymine3. Adenine
Retention and Peak Shape Under 100% Aqueous Conditions 10 mM Ammonium Formate, pH 3.0, 30 ˚C
ACQUITY UPLC HSS T3 columns retain compounds longer than ACQUITY UPLC BEH C18 columns.
Superior Retention of Polar Compounds
Compounds 1. Norepinephrine2. Epinephrine3. Dopamine4. 3,4- Dihydroxyphenylacetic acid (DOPAC)5. Serotonin (5-HT)6. 5-Hydroxy-3-indoleacetic acid (5-HIAA)7. 4-Hydroxy-3-methoxyphenylacetic acid (HVA)
Columns: As Indicated Mobile Phase A: 10mM CH3COONH4, pH 5.0Mobile Phase B: ACNFlow Rate: 0.438 mL/min Isocratic Mobile Phase Composition: 2% BInjection Volume: 0.7 µLTemperature: 30 ˚CDetection: UV @ 280 nmInstrument: ACQUITY UPLC System with ACQUITY UPLC 2996 PDA
1
2
34 5
7
6
1
23 4
5
7
6
0 1 2 3 4 min
0 1 2 3 4 min
Superior Low pH Stability
20 hr Exposure to 0.5% TFA at 60 ˚C
T3 bonding allows Atlantis T3 columns to provide long column lifetimes under harsh, low pH conditions.
60 70%40 5020 300 10
Atlantis T3
SunFire™ C18
Phenomenex® Synergi™ Hydro-RP
Shiseido Capcell Pak™ C18 AQ®
Phenomenex® Synergi™ Polar-RP
Agilent Zorbax® Eclipse Plus
Atlantis dC18
Phenomenex® Aqua
GL Sciences Inertsil™ ODS-SP
% Loss of Retention for Methyl Paraben
long colUMn lifEt iMEs at loW ph
The creation of highly retentive, reversed-phase C18 columns for polar compound retention involves bonding at a ligand density that is less hydrophobic and, therefore, more compatible with the weak, highly aqueous mobile phases required for retaining polar compounds. One disadvantage of this ligand bonding procedure is bonded phase hydrolysis under low pH (i.e., pH 2) operating conditions resulting in a gradual loss of compound retention, changes in selectivity and efficiency loss.
Atlantis T3 columns are uniquely and specifically designed to offer long, predictable column lifetimes under low pH conditions while also providing superior polar compound retention and peak shape.
a Univ Ersal rEv ErsEd-PhasE Polar coMPoUnd rEt Ent ion colUMn
Large organizations are looking to narrow or limit the number of column choices available to their separations scientists. This worldwide LC column “harmonization” can provide more predictable, controlled consumable costs, simplified method transfer between laboratory locations around the world and prescreened LC columns for various performance parameters.
With good retention of, and selectivity for, neutral, hydrophobic compounds as well as polar analytes and superior performance over a wider pH range, Atlantis T3 columns merit serious consideration as a more universal alternative to traditional C18 reversed-phase LC columns.
Column: Atlantis T3 4.6 x 150 mm, 5 µmMobile Phase A: H2OMobile Phase B: ACNMobile Phase C: 100 mM NH4HOOCH, pH 3.0Flow Rate: 2.0 mL/min Gradient: Time Profile (min) %A %B %C 0.0 85 5 10 3.0 65 25 10 10.0 65 25 10 11.0 90 0 10 20.0 90 0 10 Injection Volume: 10 µLTemperature: 30 ˚CDetection: UV @ 280 nm Instrument: Waters Alliance 2695, 2996 PDA
Compounds 1. Atenolol 2. Pindolol 3. Metoprolol 4. Oxeprenolol 5. Labetolol 6. Propranolol
Atlantis T3 columns do more than just retain and separate polar compounds.
1
2
3
4
5
6
2.00 4.00 6.00 8.00 10.00 12.00 14.000.00
Not Just for Polar Compounds
Improved Peak Shape for Bases at pH 7
ACQUITY UPLC BEH COLUMNS ACQUITY UPLC HSS Columns Atlantis HPLC Columns
Intended Use UPLC UPLC HPLC
Particle Type Ethylene Bridged Hybrid [BEH] High Strength Silica [HSS] Atlantis Silica
Available Chemistries C18, C8, Shield RP18, Phenyl, HILIC T3, C18, C18 SB T3; dC18; Unbonded [HILIC]
pH Range 1-12; [RP18: 2-11]; [HILIC: 1-8] T3, C18 SB: 2-8; C18: 1-8 T3: 2-8; dC18: 3-7; HILIC Silica: 1-5
Maximum Rated Pressure 15000 psi [~1000 bar] 15000 psi [~1000 bar] 6000 psi [~400 bar]
Particle Size 1.7 µm 1.8 µm 3, 5, 10 µm
Pore Diameter/Volume 130Å / 0.7 mL/g 100Å / 0.7 mL/g 100Å / 1.0 mL/g
Surface Area 185 m2/g 230 m2/g 330 m2/g
ACQUITY UPLC BEH C18
ACQUITY UPLC HSS T3
Minutes
Minutes
Minutes
As chromatographers around the world continue to realize the benefits of UPLC technology in different application areas, Waters continues to develop new UPLC particles and chemistries to meet these ever-growing needs.
ACQUITY UPLC HSS (High Strength Silica) columns contain the first and only 100% silica particle designed, tested and intended for use in applications up to 15000 psi/1000 bar. The ACQUITY UPLC HSS particle is not an HPLC particle. High pore volume HPLC particles do not posses the mechanical stability necessary to withstand the high column packing and operating pressures of UPLC technology. This rugged and unique UPLC- certified silica particle is specifically designed to withstand the demanding operation conditions encountered in UPLC separations.
Waters is the only manufacturer offering two UPLC-certified particles.
acQUit Y UPlc hss t3 colUMns for Polar coMPoUnd rEt Ent ion
ACQUITY UPLC HSS T3 columns utilize T3 bonding in order to retain and separate polar organic compounds. ACQUITY UPLC HSS T3 columns possess the superior polar compound retention, aqueous mobile phase compatibility and ultra-low MS bleed of Atlantis T3 columns.
When compared to ACQUITY UPLC BEH C18 columns, compounds are more strongly retained on ACQUITY UPLC HSS T3 columns.
Features of the ACQUITY UPLC HSS T3 columns include:
■ Superior polar and non-polar compound retention
■ Aqueous mobile phase compatibility
■ Ultra-low MS bleed
■ Additional selectivity choice for UPLC separations.
acQUit Y UPlc hss colUMnsiMP rov Ed ph 7 PErforManc E
At pH 7, poor peak shape for amine-containing bases and shortened column lifetimes are encountered when using intermediate ligand density C18 columns designed for polar compound retention. Poor peak shape is due to secondary interactions with unreacted silanols that remain present after bonding and endcapping. The proprietary T3 end-capping procedure reacts with more of these active silanols thereby dramatically improving peak shape for bases. Shortened column lifetimes are due to the dissolution of the silica particle substrate by the high pH mobile phase. The more effective and efficient T3 bonding helps “protect” the silica substrate from dissolution, thus providing longer column lifetimes.
When compared to other reversed-phase “polar compound retention columns,” Atlantis T3 columns provide superior pH 7 peak shape.
1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0
Thermo Aquasil C18
Agilent Zorbax® SB-Aq
Atlantis dC18
Phenomenex® Aqua
Phenomenex Synergi™ Hydro-RP
Phenomenex Synergi™ Polar-RP
Shiseido Capcell PAK™ C18 AQ®
GL Sciences Inertsil™ ODS-SP
Atlantis T3
Amitriptyline USP Tailing Factor
Did not elute
t3 Bonding
The superior performance of Atlantis T3 HPLC columns and the ACQUITY UPLC HSS T3 columns is a result of Waters new, advanced T3 bonding process. T3 bonding utilizes a trifunctional C18 alkyl phase bonded at a ligand density that promotes polar compound retention and aqueous mobile phase compatibility. The proprietary T3 endcapping process is much more effective than traditional trimethyl silane (TMS) endcapping.
This unique combination of bonding and endcapping provides superior polar compound retention and aqueous compatibility while also enhancing column performance, lifetime, peak shape and stability.
To learn more about T3 bonding and polar compound retention, please refer to Waters white paper entitled:
Topics in Liquid Chromatography Part 1. Designing a Reversed-Phase Column for Polar Compound Retention
Part Number 720001889EN
0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00
1 2 3 45
Atlantis T3
12
4 3,5
Agilent Zorbax® SB-Aq
1 2 4
3
5
Thermo Aquasil C18
12,4
3 5
Phenomenex® Synergi™ Polar - RP
12
Phenomenex® Synergi™ Hydro- RP
3,45
1 23 4 5
Shiseido Capcell Pak™ C18 AQ®
Atlantis T3 columns provide 100% aqueous compatibility, superior polar compound retention and excellent peak shape for acids, neutrals and bases.
Compounds 1. Thiourea 4. Guanosine - 5’- monophosphate2. 5-Fluorocytosine 5. Thymine3. Adenine
Retention and Peak Shape Under 100% Aqueous Conditions 10 mM Ammonium Formate, pH 3.0, 30 ˚C
ACQUITY UPLC HSS T3 columns retain compounds longer than ACQUITY UPLC BEH C18 columns.
Superior Retention of Polar Compounds
Compounds 1. Norepinephrine2. Epinephrine3. Dopamine4. 3,4- Dihydroxyphenylacetic acid (DOPAC)5. Serotonin (5-HT)6. 5-Hydroxy-3-indoleacetic acid (5-HIAA)7. 4-Hydroxy-3-methoxyphenylacetic acid (HVA)
Columns: As Indicated Mobile Phase A: 10mM CH3COONH4, pH 5.0Mobile Phase B: ACNFlow Rate: 0.438 mL/min Isocratic Mobile Phase Composition: 2% BInjection Volume: 0.7 µLTemperature: 30 ˚CDetection: UV @ 280 nmInstrument: ACQUITY UPLC System with ACQUITY UPLC 2996 PDA
1
2
34 5
7
6
1
23 4
5
7
6
0 1 2 3 4 min
0 1 2 3 4 min
Superior Low pH Stability
20 hr Exposure to 0.5% TFA at 60 ˚C
T3 bonding allows Atlantis T3 columns to provide long column lifetimes under harsh, low pH conditions.
60 70%40 5020 300 10
Atlantis T3
SunFire™ C18
Phenomenex® Synergi™ Hydro-RP
Shiseido Capcell Pak™ C18 AQ®
Phenomenex® Synergi™ Polar-RP
Agilent Zorbax® Eclipse Plus
Atlantis dC18
Phenomenex® Aqua
GL Sciences Inertsil™ ODS-SP
% Loss of Retention for Methyl Paraben
long colUMn lifEt iMEs at loW ph
The creation of highly retentive, reversed-phase C18 columns for polar compound retention involves bonding at a ligand density that is less hydrophobic and, therefore, more compatible with the weak, highly aqueous mobile phases required for retaining polar compounds. One disadvantage of this ligand bonding procedure is bonded phase hydrolysis under low pH (i.e., pH 2) operating conditions resulting in a gradual loss of compound retention, changes in selectivity and efficiency loss.
Atlantis T3 columns are uniquely and specifically designed to offer long, predictable column lifetimes under low pH conditions while also providing superior polar compound retention and peak shape.
a Univ Ersal rEv ErsEd-PhasE Polar coMPoUnd rEt Ent ion colUMn
Large organizations are looking to narrow or limit the number of column choices available to their separations scientists. This worldwide LC column “harmonization” can provide more predictable, controlled consumable costs, simplified method transfer between laboratory locations around the world and prescreened LC columns for various performance parameters.
With good retention of, and selectivity for, neutral, hydrophobic compounds as well as polar analytes and superior performance over a wider pH range, Atlantis T3 columns merit serious consideration as a more universal alternative to traditional C18 reversed-phase LC columns.
Column: Atlantis T3 4.6 x 150 mm, 5 µmMobile Phase A: H2OMobile Phase B: ACNMobile Phase C: 100 mM NH4HOOCH, pH 3.0Flow Rate: 2.0 mL/min Gradient: Time Profile (min) %A %B %C 0.0 85 5 10 3.0 65 25 10 10.0 65 25 10 11.0 90 0 10 20.0 90 0 10 Injection Volume: 10 µLTemperature: 30 ˚CDetection: UV @ 280 nm Instrument: Waters Alliance 2695, 2996 PDA
Compounds 1. Atenolol 2. Pindolol 3. Metoprolol 4. Oxeprenolol 5. Labetolol 6. Propranolol
Atlantis T3 columns do more than just retain and separate polar compounds.
1
2
3
4
5
6
2.00 4.00 6.00 8.00 10.00 12.00 14.000.00
Not Just for Polar Compounds
Improved Peak Shape for Bases at pH 7
ACQUITY UPLC BEH COLUMNS ACQUITY UPLC HSS Columns Atlantis HPLC Columns
Intended Use UPLC UPLC HPLC
Particle Type Ethylene Bridged Hybrid [BEH] High Strength Silica [HSS] Atlantis Silica
Available Chemistries C18, C8, Shield RP18, Phenyl, HILIC T3, C18, C18 SB T3; dC18; Unbonded [HILIC]
pH Range 1-12; [RP18: 2-11]; [HILIC: 1-8] T3, C18 SB: 2-8; C18: 1-8 T3: 2-8; dC18: 3-7; HILIC Silica: 1-5
Maximum Rated Pressure 15000 psi [~1000 bar] 15000 psi [~1000 bar] 6000 psi [~400 bar]
Particle Size 1.7 µm 1.8 µm 3, 5, 10 µm
Pore Diameter/Volume 130Å / 0.7 mL/g 100Å / 0.7 mL/g 100Å / 1.0 mL/g
Surface Area 185 m2/g 230 m2/g 330 m2/g
ACQUITY UPLC BEH C18
ACQUITY UPLC HSS T3
Minutes
Minutes
Minutes
As chromatographers around the world continue to realize the benefits of UPLC technology in different application areas, Waters continues to develop new UPLC particles and chemistries to meet these ever-growing needs.
ACQUITY UPLC HSS (High Strength Silica) columns contain the first and only 100% silica particle designed, tested and intended for use in applications up to 15000 psi/1000 bar. The ACQUITY UPLC HSS particle is not an HPLC particle. High pore volume HPLC particles do not posses the mechanical stability necessary to withstand the high column packing and operating pressures of UPLC technology. This rugged and unique UPLC- certified silica particle is specifically designed to withstand the demanding operation conditions encountered in UPLC separations.
Waters is the only manufacturer offering two UPLC-certified particles.
acQUit Y UPlc hss t3 colUMns for Polar coMPoUnd rEt Ent ion
ACQUITY UPLC HSS T3 columns utilize T3 bonding in order to retain and separate polar organic compounds. ACQUITY UPLC HSS T3 columns possess the superior polar compound retention, aqueous mobile phase compatibility and ultra-low MS bleed of Atlantis T3 columns.
When compared to ACQUITY UPLC BEH C18 columns, compounds are more strongly retained on ACQUITY UPLC HSS T3 columns.
Features of the ACQUITY UPLC HSS T3 columns include:
■ Superior polar and non-polar compound retention
■ Aqueous mobile phase compatibility
■ Ultra-low MS bleed
■ Additional selectivity choice for UPLC separations.
acQUit Y UPlc hss colUMnsiMP rov Ed ph 7 PErforManc E
At pH 7, poor peak shape for amine-containing bases and shortened column lifetimes are encountered when using intermediate ligand density C18 columns designed for polar compound retention. Poor peak shape is due to secondary interactions with unreacted silanols that remain present after bonding and endcapping. The proprietary T3 end-capping procedure reacts with more of these active silanols thereby dramatically improving peak shape for bases. Shortened column lifetimes are due to the dissolution of the silica particle substrate by the high pH mobile phase. The more effective and efficient T3 bonding helps “protect” the silica substrate from dissolution, thus providing longer column lifetimes.
When compared to other reversed-phase “polar compound retention columns,” Atlantis T3 columns provide superior pH 7 peak shape.
1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0
Thermo Aquasil C18
Agilent Zorbax® SB-Aq
Atlantis dC18
Phenomenex® Aqua
Phenomenex Synergi™ Hydro-RP
Phenomenex Synergi™ Polar-RP
Shiseido Capcell PAK™ C18 AQ®
GL Sciences Inertsil™ ODS-SP
Atlantis T3
Amitriptyline USP Tailing Factor
Did not elute
t3 Bonding
The superior performance of Atlantis T3 HPLC columns and the ACQUITY UPLC HSS T3 columns is a result of Waters new, advanced T3 bonding process. T3 bonding utilizes a trifunctional C18 alkyl phase bonded at a ligand density that promotes polar compound retention and aqueous mobile phase compatibility. The proprietary T3 endcapping process is much more effective than traditional trimethyl silane (TMS) endcapping.
This unique combination of bonding and endcapping provides superior polar compound retention and aqueous compatibility while also enhancing column performance, lifetime, peak shape and stability.
To learn more about T3 bonding and polar compound retention, please refer to Waters white paper entitled:
Topics in Liquid Chromatography Part 1. Designing a Reversed-Phase Column for Polar Compound Retention
Part Number 720001889EN
0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00
1 2 3 45
Atlantis T3
12
4 3,5
Agilent Zorbax® SB-Aq
1 2 4
3
5
Thermo Aquasil C18
12,4
3 5
Phenomenex® Synergi™ Polar - RP
12
Phenomenex® Synergi™ Hydro- RP
3,45
1 23 4 5
Shiseido Capcell Pak™ C18 AQ®
Atlantis T3 columns provide 100% aqueous compatibility, superior polar compound retention and excellent peak shape for acids, neutrals and bases.
Compounds 1. Thiourea 4. Guanosine - 5’- monophosphate2. 5-Fluorocytosine 5. Thymine3. Adenine
Retention and Peak Shape Under 100% Aqueous Conditions 10 mM Ammonium Formate, pH 3.0, 30 ˚C
ACQUITY UPLC HSS T3 columns retain compounds longer than ACQUITY UPLC BEH C18 columns.
Superior Retention of Polar Compounds
Compounds 1. Norepinephrine2. Epinephrine3. Dopamine4. 3,4- Dihydroxyphenylacetic acid (DOPAC)5. Serotonin (5-HT)6. 5-Hydroxy-3-indoleacetic acid (5-HIAA)7. 4-Hydroxy-3-methoxyphenylacetic acid (HVA)
Columns: As Indicated Mobile Phase A: 10mM CH3COONH4, pH 5.0Mobile Phase B: ACNFlow Rate: 0.438 mL/min Isocratic Mobile Phase Composition: 2% BInjection Volume: 0.7 µLTemperature: 30 ˚CDetection: UV @ 280 nmInstrument: ACQUITY UPLC System with ACQUITY UPLC 2996 PDA
1
2
34 5
7
6
1
23 4
5
7
6
0 1 2 3 4 min
0 1 2 3 4 min
Superior Low pH Stability
20 hr Exposure to 0.5% TFA at 60 ˚C
T3 bonding allows Atlantis T3 columns to provide long column lifetimes under harsh, low pH conditions.
60 70%40 5020 300 10
Atlantis T3
SunFire™ C18
Phenomenex® Synergi™ Hydro-RP
Shiseido Capcell Pak™ C18 AQ®
Phenomenex® Synergi™ Polar-RP
Agilent Zorbax® Eclipse Plus
Atlantis dC18
Phenomenex® Aqua
GL Sciences Inertsil™ ODS-SP
% Loss of Retention for Methyl Paraben
long colUMn lifEt iMEs at loW ph
The creation of highly retentive, reversed-phase C18 columns for polar compound retention involves bonding at a ligand density that is less hydrophobic and, therefore, more compatible with the weak, highly aqueous mobile phases required for retaining polar compounds. One disadvantage of this ligand bonding procedure is bonded phase hydrolysis under low pH (i.e., pH 2) operating conditions resulting in a gradual loss of compound retention, changes in selectivity and efficiency loss.
Atlantis T3 columns are uniquely and specifically designed to offer long, predictable column lifetimes under low pH conditions while also providing superior polar compound retention and peak shape.
a Univ Ersal rEv ErsEd-PhasE Polar coMPoUnd rEt Ent ion colUMn
Large organizations are looking to narrow or limit the number of column choices available to their separations scientists. This worldwide LC column “harmonization” can provide more predictable, controlled consumable costs, simplified method transfer between laboratory locations around the world and prescreened LC columns for various performance parameters.
With good retention of, and selectivity for, neutral, hydrophobic compounds as well as polar analytes and superior performance over a wider pH range, Atlantis T3 columns merit serious consideration as a more universal alternative to traditional C18 reversed-phase LC columns.
Column: Atlantis T3 4.6 x 150 mm, 5 µmMobile Phase A: H2OMobile Phase B: ACNMobile Phase C: 100 mM NH4HOOCH, pH 3.0Flow Rate: 2.0 mL/min Gradient: Time Profile (min) %A %B %C 0.0 85 5 10 3.0 65 25 10 10.0 65 25 10 11.0 90 0 10 20.0 90 0 10 Injection Volume: 10 µLTemperature: 30 ˚CDetection: UV @ 280 nm Instrument: Waters Alliance 2695, 2996 PDA
Compounds 1. Atenolol 2. Pindolol 3. Metoprolol 4. Oxeprenolol 5. Labetolol 6. Propranolol
Atlantis T3 columns do more than just retain and separate polar compounds.
1
2
3
4
5
6
2.00 4.00 6.00 8.00 10.00 12.00 14.000.00
Not Just for Polar Compounds
Improved Peak Shape for Bases at pH 7
ACQUITY UPLC BEH COLUMNS ACQUITY UPLC HSS Columns Atlantis HPLC Columns
Intended Use UPLC UPLC HPLC
Particle Type Ethylene Bridged Hybrid [BEH] High Strength Silica [HSS] Atlantis Silica
Available Chemistries C18, C8, Shield RP18, Phenyl, HILIC T3, C18, C18 SB T3; dC18; Unbonded [HILIC]
pH Range 1-12; [RP18: 2-11]; [HILIC: 1-8] T3, C18 SB: 2-8; C18: 1-8 T3: 2-8; dC18: 3-7; HILIC Silica: 1-5
Maximum Rated Pressure 15000 psi [~1000 bar] 15000 psi [~1000 bar] 6000 psi [~400 bar]
Particle Size 1.7 µm 1.8 µm 3, 5, 10 µm
Pore Diameter/Volume 130Å / 0.7 mL/g 100Å / 0.7 mL/g 100Å / 1.0 mL/g
Surface Area 185 m2/g 230 m2/g 330 m2/g
ACQUITY UPLC BEH C18
ACQUITY UPLC HSS T3
Minutes
Minutes
Minutes
EasiEr MEt hod t ransfEr
Since Atlantis T3 HPLC and ACQUITY UPLC HSS T3 columns utilize T3 bonding, HPLC methods can be transferred to UPLC methods. Because Atlantis and HSS particles are different substrates, some method modification and/or optimization may be required. A compelling benefit of UPLC technology is the ease and speed at which this method optimization can occur since UPLC technology offers greater resolution in less time.
Moving from XBridge™ HPLC to ACQUITY UPLC BEH columns is nearly seamless, as the hybrid particle substrates differ only in particle size. However, because of T3 bonding it should be straightforward to make simple adjustments to successfully transfer a separation from an Atlantis T3 HPLC column to an ACQUITY UPLC HSS T3 column.
additional choicEs for UPlc sEParations
Chromatographers continue to embrace UPLC technology as the separation technology of the future. Additional UPLC column chemistries that provide complementary selectivities are needed to provide application-specific solutions. When combined with the ultra-resolution of UPLC, these new UPLC column chemistries will enable the development of fast, robust methods in less time. ACQUITY UPLC HSS T3 columns offer another selectivity choice for UPLC separations.
T3 Bonding Enables HPLC to UPLC Transfer
HPLC Separation on Atlantis T3
UPLC Separation on ACQUITY UPLC HSS T3
CompoundRetention Time (min)
USP Resolution
1. Cytosine 2.77
2. 5-Fluorocytosine 3.58 4.56
3. Uracil 3.93 1.71
4. 5-Fluorouracil 4.59 3.41
5. Guanine 5.92 7.76
6. Thymine 7.63 11.01
7. Adenine 8.39 4.70
Similar Selectivities
CompoundRetention Time (min)
USP Resolution
1. Cytosine 0.34
2. 5-Fluorocytosine 0.44 5.02
3. Uracil 0.48 1.50
4. 5-Fluorouracil 0.56 3.17
5. Guanine 0.80 8.16
6. Thymine 1.25 14.65
7. Adenine 1.33 3.50
T3 Bonding Enables HPLC to UPLC™ Transfer
HPLC on Atlantis® T3
UPLC™ on ACQUITY UPLC™ HSS T3
>80% Reduction in Run Time
0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 min
1 2 3 4 5 6 7 8 9 min
1
2
3
4
5
6
7
1
2 3
4
5
6
7
1 Cytosine2 5-Fluorocytosine3 Uracil4 5-Fluorouracil5 Guanine6 Thymine7 Adenine
T3 Bonding Enables HPLC to UPLC™ Transfer
HPLC on Atlantis® T3
UPLC™ on ACQUITY UPLC™ HSS T3
>80% Reduction in Run Time
0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 min
1 2 3 4 5 6 7 8 9 min
1
2
3
4
5
6
7
1
2 3
4
5
6
7
1 Cytosine2 5-Fluorocytosine3 Uracil4 5-Fluorouracil5 Guanine6 Thymine7 Adenine
T3 bonding allows for the transfer of HPLC methods to UPLC.
Part Number Particle Size Dimension
186003713 3 µm 1.0 x 50 mm
186003730 5 µm 1.0 x 50 mm
186003714 3 µm 1.0 x 150 mm
186003731 5 µm 1.0 x 150 mm
186003715 3 µm 2.1 x 20 mm IS™
186003732 5 µm 2.1 x 20 mm IS
186003716 3 µm 2.1 x 30 mm
186003733 5 µm 2.1 x 30 mm
186003717 3 µm 2.1 x 50 mm
186003734 5 µm 2.1 x 50 mm
186003718 3 µm 2.1 x 100 mm
186003735 5 µm 2.1 x 100 mm
186003719 3 µm 2.1 x 150 mm
186003736 5 µm 2.1 x 150 mm
186003720 3 µm 3.0 x 20 mm IS
186003737 5 µm 3.0 x 20 mm IS
186003721 3 µm 3.0 x 50 mm
186003738 5 µm 3.0 x 50 mm
186003722 3 µm 3.0 x 100 mm
186003739 5 µm 3.0 x 100 mm
186003723 3 µm 3.0 x 150 mm
186003740 5 µm 3.0 x 150 mm
186003741 5 µm 3.0 x 250 mm
186003724 3 µm 4.6 x 20 mm IS
186003742 5 µm 4.6 x 20 mm IS
186003725 3 µm 4.6 x 30 mm
186003743 5 µm 4.6 x 30 mm
186003726 3 µm 4.6 x 50 mm
186003744 5 µm 4.6 x 50 mm
186003727 3 µm 4.6 x 75 mm
186003745 5 µm 4.6 x 75 mm
186003728 3 µm 4.6 x 100 mm
186003746 5 µm 4.6 x 100 mm
186003729 3 µm 4.6 x 150 mm
186003747 5 µm 4.6 x 150 mm
186003748 5 µm 4.6 x 250 mm
Part Number Particle Size Dimension
1860036953 5 µm 10 x 10 mm Guard
1860037063 10 µm 10 x 10 mm Guard
186003691 5 µm 10 x 50 mm
186003692 5 µm 10 x 100 mm
186003693 5 µm 10 x 150 mm
186003704 10 µm 10 x 150 mm
186003694 5 µm 10 x 250 mm
186003705 10 µm 10 x 250 mm
1860036994 5 µm 19 x 10 mm Guard
1860037104 10 µm 19 x 10 mm Guard
186003696 5 µm 19 x 50 mm OBD™
186003707 10 µm 19 x 50 mm OBD
186003697 5 µm 19 x 100 mm OBD
186003698 5 µm 19 x 150 mm OBD
186003708 10 µm 19 x 150 mm OBD
186003709 10 µm 19 x 250 mm OBD
186004026 5 µm 19 x 250 mm OBD
186003700 5 µm 30 x 50 mm OBD
186003701 5 µm 30 x 75 mm OBD
186003702 5 µm 30 x 100 mm OBD
186003703 5 µm 30 x 150 mm OBD
186003711 10 µm 30 x 150 mm OBD
186003712 10 µm 30 x 250 mm OBD
186004080 5 µm 50 x 50 mm OBD
186004081 5 µm 50 x 100 mm OBD
186004082 5 µm 50 x 150 mm OBD
186004083 10 µm 50 x 50 mm OBD
186004084 10 µm 50 x 100 mm OBD
186004085 10 µm 50 x 150 mm OBD
186004086 10 µm 50 x 250 mm OBD
Part Number Particle Size Dimension
186003751 3 µm 4.6 x 150 mm
186003754 5 µm 4.6 x 150 mm
186003755 5 µm 4.6 x 250 mm
Part Number Particle Size Dimension
1860037561 3 µm 2.1 x 10 mm (2/pk)
1860037591 5 µm 2.1 x 10 mm (2/pk)
1860037572 3 µm 3.9 x 20 mm (2/pk)
1860037602 5 µm 3.9 x 20 mm (2/pk)
1860037582 3 µm 4.6 x 20 mm (2/pk)
1860037612 5 µm 4.6 x 20 mm (2/pk)
ordEring inforMat ion
1 - Requires Sentry Guard Holder WAT0979582 - Requires Sentry Guard Holder WAT0469103 - Requires Cartridge Guard Holder 2890007794 - Requires Cartridge Guard Holder 186000709
Compounds 1. Caftaric acid2. Chlorogenic acid3. Cynarin4. Echinacoside5. Cichoric acid
Columns: ACQUITY UPLC HSS 2.1 x 50 mm, 1.8 µm ACQUITY UPLC BEH 2.1 x 50 mm, 1.7 µmMobile Phase A: 0.1% CF3COOH in H2OMobile Phase B: 0.08% CF3COOH in ACNFlow Rate: 0.5 mL/min Gradient: Time Profile Curve (min) %A %B 0.0 92 8 6 0.1 92 8 6 4.45 50 50 7 4.86 10 90 6 5.0 92 8 6 6.0 92 8 6Injection Volume: 1.0 µLSample Diluent: 50:50 H2O: MeOH with 0.05% CF3COOH Sample Conc.: 100 µg/mLTemperature: 40 ˚CDetection: UV @ 330 nmSampling Rate: 40 pts/secTime Constant: 0.1Instrument: Waters ACQUITY UPLC system with ACQUITY UPLC TUV
Minutes
0.00 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00
1 24 53
BEH C18
BEH C8
BEH Shield RP18
BEH Phenyl
1 24 53
1 24 53
1 2 453
1
2 4 53HSS T3
HSS C18143 52
1 2 45
3 HSS C18 SB
The additional UPLC column selectivity choices provide and facilitate rapid development of robust methods.
Reversed-Phase UPLC Chemistry Choices: Caffeic Acid Derivatives in Echinacea
Part Number Particle Size Dimension
186004055 1.8 µm 2.1 x 50 mm
186004056 1.8 µm 2.1 x 100 mm
■ Superior polar compound retention
■ Wide usable silica pH range
■ Designed for aqueous mobile phases
■ Long column lifetimes
■ Fully Mass Spectrometry (MS) compatible
AtLAntiS t3 AnD ACQUitY UPLC HSS t3 CoLUMnS
Atlantis® T3 and ACQUITY UPLC® HSS T3 columns are the new industry standards for retaining and separating polar compounds via reversed-phase HPLC and UPLC® technology. Atlantis T3 HPLC columns are universal silica-based, reversed-phase C18 columns that not only retain and separate highly polar compounds, but also provide superior performance across a wider pH range. ACQUITY UPLC HSS T3 columns maintain all the advantages of Atlantis T3 HPLC columns but are also certified for use in UPLC separations while providing an additional UPLC chemistry choice.
The Atlantis T3 columns are designed to provide longer column lifetimes at low pH as well as improved peak shape for amine-containing bases at pH 7.
Waters has created a second pressure-tolerant, UPLC-certified particle. This High Strength Silica (HSS) particle is the first and only 100% silica particle designed specifically for 15000 psi/1000 bar applications. ACQUITY UPLC HSS T3 columns utilize the same T3 bonding and provides similar attributes as Atlantis T3 HPLC columns.
t3: rEdEfining Polar coMPoUnd rEt Ent ion in hPlc and UPlc
80% Run Time Reduction!!
Atlantis T3 Analytical Columns Atlantis T3 Preparative Columns Atlantis T3 Sentry Guard Cartridges
Atlantis T3 Method Validation Kits*
ACQUITY UPLC HSS T3 Columns
ACQUITY UPLC HSS T3 Method Validation Kits*
* Contains 3 columns, each packed with a different batch of packing material.
Waters corporation 34 Maple Street Milford, MA 01757 U.S.A. T: 1 508 478 2000 F: 1 508 872 1990 www.waters.com
Waters, The Science of What’s Possible, Atlantis, ACQUITY UPLC, UPLC, ACQUITY, ACQUITY Ultra Performance LC, SunFire, XBridge, IS, OBD, Alliance and Sentry are trademarks of Waters Corporation. All trademarks used are acknowledged. All rights reserved.All other trademarks are the property of their respective owners.
©2007 Waters Corporation. Printed/Produced in the U.S.A.November 2007 720001887EN SC-XX
Part Number Particle Size Dimension
186003535 1.8 µm 1.0 x 50 mm
186003536 1.8 µm 1.0 x 100 mm
186003537 1.8 µm 1.0 x 150 mm
186003944 1.8 µm 2.1 x 30 mm
186003538 1.8 µm 2.1 x 50 mm
186003539 1.8 µm 2.1 x 100 mm
186003540 1.8 µm 2.1 x 150 mm
Minutes
Minutes
EasiEr MEt hod t ransfEr
Since Atlantis T3 HPLC and ACQUITY UPLC HSS T3 columns utilize T3 bonding, HPLC methods can be transferred to UPLC methods. Because Atlantis and HSS particles are different substrates, some method modification and/or optimization may be required. A compelling benefit of UPLC technology is the ease and speed at which this method optimization can occur since UPLC technology offers greater resolution in less time.
Moving from XBridge™ HPLC to ACQUITY UPLC BEH columns is nearly seamless, as the hybrid particle substrates differ only in particle size. However, because of T3 bonding it should be straightforward to make simple adjustments to successfully transfer a separation from an Atlantis T3 HPLC column to an ACQUITY UPLC HSS T3 column.
additional choicEs for UPlc sEParations
Chromatographers continue to embrace UPLC technology as the separation technology of the future. Additional UPLC column chemistries that provide complementary selectivities are needed to provide application-specific solutions. When combined with the ultra-resolution of UPLC, these new UPLC column chemistries will enable the development of fast, robust methods in less time. ACQUITY UPLC HSS T3 columns offer another selectivity choice for UPLC separations.
T3 Bonding Enables HPLC to UPLC Transfer
HPLC Separation on Atlantis T3
UPLC Separation on ACQUITY UPLC HSS T3
CompoundRetention Time (min)
USP Resolution
1. Cytosine 2.77
2. 5-Fluorocytosine 3.58 4.56
3. Uracil 3.93 1.71
4. 5-Fluorouracil 4.59 3.41
5. Guanine 5.92 7.76
6. Thymine 7.63 11.01
7. Adenine 8.39 4.70
Similar Selectivities
CompoundRetention Time (min)
USP Resolution
1. Cytosine 0.34
2. 5-Fluorocytosine 0.44 5.02
3. Uracil 0.48 1.50
4. 5-Fluorouracil 0.56 3.17
5. Guanine 0.80 8.16
6. Thymine 1.25 14.65
7. Adenine 1.33 3.50
T3 Bonding Enables HPLC to UPLC™ Transfer
HPLC on Atlantis® T3
UPLC™ on ACQUITY UPLC™ HSS T3
>80% Reduction in Run Time
0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 min
1 2 3 4 5 6 7 8 9 min
1
2
3
4
5
6
7
1
2 3
4
5
6
7
1 Cytosine2 5-Fluorocytosine3 Uracil4 5-Fluorouracil5 Guanine6 Thymine7 Adenine
T3 Bonding Enables HPLC to UPLC™ Transfer
HPLC on Atlantis® T3
UPLC™ on ACQUITY UPLC™ HSS T3
>80% Reduction in Run Time
0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 min
1 2 3 4 5 6 7 8 9 min
1
2
3
4
5
6
7
1
2 3
4
5
6
7
1 Cytosine2 5-Fluorocytosine3 Uracil4 5-Fluorouracil5 Guanine6 Thymine7 Adenine
T3 bonding allows for the transfer of HPLC methods to UPLC.
Part Number Particle Size Dimension
186003713 3 µm 1.0 x 50 mm
186003730 5 µm 1.0 x 50 mm
186003714 3 µm 1.0 x 150 mm
186003731 5 µm 1.0 x 150 mm
186003715 3 µm 2.1 x 20 mm IS™
186003732 5 µm 2.1 x 20 mm IS
186003716 3 µm 2.1 x 30 mm
186003733 5 µm 2.1 x 30 mm
186003717 3 µm 2.1 x 50 mm
186003734 5 µm 2.1 x 50 mm
186003718 3 µm 2.1 x 100 mm
186003735 5 µm 2.1 x 100 mm
186003719 3 µm 2.1 x 150 mm
186003736 5 µm 2.1 x 150 mm
186003720 3 µm 3.0 x 20 mm IS
186003737 5 µm 3.0 x 20 mm IS
186003721 3 µm 3.0 x 50 mm
186003738 5 µm 3.0 x 50 mm
186003722 3 µm 3.0 x 100 mm
186003739 5 µm 3.0 x 100 mm
186003723 3 µm 3.0 x 150 mm
186003740 5 µm 3.0 x 150 mm
186003741 5 µm 3.0 x 250 mm
186003724 3 µm 4.6 x 20 mm IS
186003742 5 µm 4.6 x 20 mm IS
186003725 3 µm 4.6 x 30 mm
186003743 5 µm 4.6 x 30 mm
186003726 3 µm 4.6 x 50 mm
186003744 5 µm 4.6 x 50 mm
186003727 3 µm 4.6 x 75 mm
186003745 5 µm 4.6 x 75 mm
186003728 3 µm 4.6 x 100 mm
186003746 5 µm 4.6 x 100 mm
186003729 3 µm 4.6 x 150 mm
186003747 5 µm 4.6 x 150 mm
186003748 5 µm 4.6 x 250 mm
Part Number Particle Size Dimension
1860036953 5 µm 10 x 10 mm Guard
1860037063 10 µm 10 x 10 mm Guard
186003691 5 µm 10 x 50 mm
186003692 5 µm 10 x 100 mm
186003693 5 µm 10 x 150 mm
186003704 10 µm 10 x 150 mm
186003694 5 µm 10 x 250 mm
186003705 10 µm 10 x 250 mm
1860036994 5 µm 19 x 10 mm Guard
1860037104 10 µm 19 x 10 mm Guard
186003696 5 µm 19 x 50 mm OBD™
186003707 10 µm 19 x 50 mm OBD
186003697 5 µm 19 x 100 mm OBD
186003698 5 µm 19 x 150 mm OBD
186003708 10 µm 19 x 150 mm OBD
186003709 10 µm 19 x 250 mm OBD
186004026 5 µm 19 x 250 mm OBD
186003700 5 µm 30 x 50 mm OBD
186003701 5 µm 30 x 75 mm OBD
186003702 5 µm 30 x 100 mm OBD
186003703 5 µm 30 x 150 mm OBD
186003711 10 µm 30 x 150 mm OBD
186003712 10 µm 30 x 250 mm OBD
186004080 5 µm 50 x 50 mm OBD
186004081 5 µm 50 x 100 mm OBD
186004082 5 µm 50 x 150 mm OBD
186004083 10 µm 50 x 50 mm OBD
186004084 10 µm 50 x 100 mm OBD
186004085 10 µm 50 x 150 mm OBD
186004086 10 µm 50 x 250 mm OBD
Part Number Particle Size Dimension
186003751 3 µm 4.6 x 150 mm
186003754 5 µm 4.6 x 150 mm
186003755 5 µm 4.6 x 250 mm
Part Number Particle Size Dimension
1860037561 3 µm 2.1 x 10 mm (2/pk)
1860037591 5 µm 2.1 x 10 mm (2/pk)
1860037572 3 µm 3.9 x 20 mm (2/pk)
1860037602 5 µm 3.9 x 20 mm (2/pk)
1860037582 3 µm 4.6 x 20 mm (2/pk)
1860037612 5 µm 4.6 x 20 mm (2/pk)
ordEring inforMat ion
1 - Requires Sentry Guard Holder WAT0979582 - Requires Sentry Guard Holder WAT0469103 - Requires Cartridge Guard Holder 2890007794 - Requires Cartridge Guard Holder 186000709
Compounds 1. Caftaric acid2. Chlorogenic acid3. Cynarin4. Echinacoside5. Cichoric acid
Columns: ACQUITY UPLC HSS 2.1 x 50 mm, 1.8 µm ACQUITY UPLC BEH 2.1 x 50 mm, 1.7 µmMobile Phase A: 0.1% CF3COOH in H2OMobile Phase B: 0.08% CF3COOH in ACNFlow Rate: 0.5 mL/min Gradient: Time Profile Curve (min) %A %B 0.0 92 8 6 0.1 92 8 6 4.45 50 50 7 4.86 10 90 6 5.0 92 8 6 6.0 92 8 6Injection Volume: 1.0 µLSample Diluent: 50:50 H2O: MeOH with 0.05% CF3COOH Sample Conc.: 100 µg/mLTemperature: 40 ˚CDetection: UV @ 330 nmSampling Rate: 40 pts/secTime Constant: 0.1Instrument: Waters ACQUITY UPLC system with ACQUITY UPLC TUV
Minutes
0.00 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00
1 24 53
BEH C18
BEH C8
BEH Shield RP18
BEH Phenyl
1 24 53
1 24 53
1 2 453
1
2 4 53HSS T3
HSS C18143 52
1 2 45
3 HSS C18 SB
The additional UPLC column selectivity choices provide and facilitate rapid development of robust methods.
Reversed-Phase UPLC Chemistry Choices: Caffeic Acid Derivatives in Echinacea
Part Number Particle Size Dimension
186004055 1.8 µm 2.1 x 50 mm
186004056 1.8 µm 2.1 x 100 mm
■ Superior polar compound retention
■ Wide usable silica pH range
■ Designed for aqueous mobile phases
■ Long column lifetimes
■ Fully Mass Spectrometry (MS) compatible
AtLAntiS t3 AnD ACQUitY UPLC HSS t3 CoLUMnS
Atlantis® T3 and ACQUITY UPLC® HSS T3 columns are the new industry standards for retaining and separating polar compounds via reversed-phase HPLC and UPLC® technology. Atlantis T3 HPLC columns are universal silica-based, reversed-phase C18 columns that not only retain and separate highly polar compounds, but also provide superior performance across a wider pH range. ACQUITY UPLC HSS T3 columns maintain all the advantages of Atlantis T3 HPLC columns but are also certified for use in UPLC separations while providing an additional UPLC chemistry choice.
The Atlantis T3 columns are designed to provide longer column lifetimes at low pH as well as improved peak shape for amine-containing bases at pH 7.
Waters has created a second pressure-tolerant, UPLC-certified particle. This High Strength Silica (HSS) particle is the first and only 100% silica particle designed specifically for 15000 psi/1000 bar applications. ACQUITY UPLC HSS T3 columns utilize the same T3 bonding and provides similar attributes as Atlantis T3 HPLC columns.
t3: rEdEfining Polar coMPoUnd rEt Ent ion in hPlc and UPlc
80% Run Time Reduction!!
Atlantis T3 Analytical Columns Atlantis T3 Preparative Columns Atlantis T3 Sentry Guard Cartridges
Atlantis T3 Method Validation Kits*
ACQUITY UPLC HSS T3 Columns
ACQUITY UPLC HSS T3 Method Validation Kits*
* Contains 3 columns, each packed with a different batch of packing material.
Waters corporation 34 Maple Street Milford, MA 01757 U.S.A. T: 1 508 478 2000 F: 1 508 872 1990 www.waters.com
Waters, The Science of What’s Possible, Atlantis, ACQUITY UPLC, UPLC, ACQUITY, ACQUITY Ultra Performance LC, SunFire, XBridge, IS, OBD, Alliance and Sentry are trademarks of Waters Corporation. All trademarks used are acknowledged. All rights reserved.All other trademarks are the property of their respective owners.
©2007 Waters Corporation. Printed/Produced in the U.S.A.November 2007 720001887EN SC-XX
Part Number Particle Size Dimension
186003535 1.8 µm 1.0 x 50 mm
186003536 1.8 µm 1.0 x 100 mm
186003537 1.8 µm 1.0 x 150 mm
186003944 1.8 µm 2.1 x 30 mm
186003538 1.8 µm 2.1 x 50 mm
186003539 1.8 µm 2.1 x 100 mm
186003540 1.8 µm 2.1 x 150 mm
Minutes
Minutes
