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Thermo Scientific Accucore HPLC Columns

Stephen LukeProduct ManagerJune 2011

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Fast Chromatography - Understanding the Drivers

• Analytical Scientist want

FAST CHROMATOGRAPHY

Speed 62%

Resolution 23%

Sensitivity 7%

Other 8%

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Fast HPLC – Particle Evolution

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Core Enhanced TechnologyTM

Solid Core Particles – 2.6µm diameter particles with a solid core generate high speed, high resolution separations without excessive backpressure

Advanced Bonding Technology – Optimised phase bonding creates a series of high coverage, robust phases

Tight Control of Particle Diameter – Enhanced selection process keeps particle size distribution to a minimum and produces high efficiency columns

Automated Packing Process -Enhanced automated procedures ensure that all columns are packed with the highest quality

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0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0Hei

ght E

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t The

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ate

Linear velocity of mobile phase (mm/s)Accucore 2.6µm 5µm 3µm <2µm

Core Enhanced Technology - Efficiency

Highest efficiency and lowest rate of efficiency loss with flow rate with Core Enhanced Technology

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0.0003

0.0030

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1,000.00 10,000.00 100,000.00 1,000,000.00

Accucore RP-MS 2.6um 5um 3um <2umN (/)

Nopt

t0/N (s)

NoptTim

e re

quire

d to

obt

ain

a se

t effi

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Core Enhanced Technology – Efficiency & Speed

Efficiency increasing

Highest plate generation rate with Core Enhanced Technology

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Core Enhanced Technology – Low Pressure

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Accucore RP-MS 2.6µm <2µm 3µm 5µm

Columns: 100 x 2.1 mmMobile phase: H2O / ACN (1:1)Temperature: 30 °C

600 bar limit

HPLC pressure limit

Pressures below 600 bar over optimum flow rate range with Core Enhanced Technology

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• Ultimate Core Performance to Maximize Your Investment

• Rugged and reproducible solid core particles• Fast separations with superb resolution• Low backpressures

AccucoreTM HPLC Columns

Core Enhanced Technology

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Accucore HPLC Columns – Features and Benefits

Ultimate Core Performance Maximizing Your Investment

Fast Separations Increase sample throughput and decrease solvent costs

High Peak Capacity Separate and identify more peaks

Increased Sensitivity Detect trace amounts

Lower Pressure Use existing HPLC instruments or prolong the life of UHPLC instruments

Loading Capacity Analyze a wide range of concentrations

Reproducible Chromatography Have confidence in your separations

Long Lifetime Use columns for longer

Wide Selectivity Use the right column for your analysis

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Accucore HPLC Columns - Specifications

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Accucore HPLC Columns - Characterization

Hydrophobic Interactions

Secondary Interactions

Acidic Interactions

Hydrophobic retention

Base activity Acid interaction

HR BA AI Hydrophobic selectivity

Chelation

Ion exchange capacity pH 2.7

HS C IEX (2.7) Steric selectivity Ion exchange

capacity (pH 7.6)

SS IEX (7.6) Hydrogen bonding capacity

HBC

HR K’ Pentylbenzene

HS α Butylbenzene / Pentylbenzene

SS α Triphenylene / o-Terphenyl

HBC α Caffeine / Phenol

BA tf Amitripyline

C tf Quinizarin

IEX(7.6) α Benzylamine / Phenol

AI tf 4-Chlorocinnamic acid

IEX(2.7) α Benzylamine / Phenol

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Accucore HPLC Columns – Phase Range

RP-MS Optimized for MS detection, excellent combination of speed and quality of separation

Phenyl-Hexyl Unique selectivity for aromatic and moderately polar

analytes

C18 Optimum retention for non-polar analytes

PFP Alternative

selectivity to C18, particularly for

halogenated analytes

aQ Compatible with 100% aqueous mobile phases, special selectivity for polar analytes

HILIC

Enhanced Retention of polar

and hydrophilic analytes

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Accucore RP-MS

• Optimized for MS detection• Bleed tested

• Excellent Peak Shapes• Fewer mobile phase modifiers

• Best Combination of Speed and Efficiency

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Accucore C18

• Optimum retention of non-polar compounds • Hydrophobic interaction mechanism• Separate a broad range of analytes

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Accucore aQ

• Retention and Resolution of Polar Analytes• Polar Endcapped C18 Stationary Phase for Alternative Selectivity• Ideal for Highly Aqueous Mobile Phases

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Accucore Phenyl-Hexyl

• Mixed-Mode Selectivity for Aromatic and Moderately Polar Analytes• Enhanced Pi-pi Interactions with Aromatics• Moderate Hydrophobicity

L11

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Accucore PFP

• Alternative Selectivity to C18• Extra Retention for Halogenated Species• Unique Selectivity for Non-halogenated Polar Compounds

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Accucore HILIC

• Enhanced Retention of Polar and Hydrophilic Analytes• Alternative Selectivity to C18 without Ion-pair or Derivatization• Improved Sensitivity for MS detection

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Accucore HPLC Columns – SpeedAccucore C18 2.6µm, 50 mm x 2.1 mmMobile phase A: water; B: AcetonitrileGradient: Min %B

0 400.4 950.80 95

Flow rate : 1.0 mL /minInjection volume : 1 µLTemperature: 45 °CDetection: UV at λ = 258 nm

Pressure: 113 bar

• Analytes: 1. 2-Pentanone2. Acetophenone3. 2-Heptanone4. Butyrophenone5. Hexanophenone6. Octanophenne7. Decanophenone

0.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9Time (min)

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High resolution separation 6 times faster than a 150x4.6mm, 5µm column – at low pressure

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Accucore HPLC Columns – Resolution

Accucore PFP 2.6µm, 100 mm x 2.1 mmMobile phase A : 0.1% Formic acid in waterMobile phase B: 0.1% Formic acid in AcetonitrileGradient: 15-30%B in 7 minutesFlow rate: 600 µL/minInjection volume: 2 µLTemperature: 50 °CDetection: UV at 270 nm

Pressure: 270 bar

Minutes0 2 4 6 8

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1 3,4 - Dimethoxyphenol

2 2,6 - Dimethoxyphenol

3 2,6 - Difluorophenol

4 3,5 - Dimethoxyphenol

5 2,4 - Difluorophenol

6 2,3 - Difluorophenol

7 3,4 - Difluorophenol

8 3,5 - Dimethylphenol

9 2,6 - Dimethylphenol

10 2,6 - Dichlorophenol

11 4 – Chloro-3-Methylphenol

12 4-Chloro-2-Methylphenol

13 3,4 - Dichlorophenol

14 3,5 - Dichlorophenol

Baseline resolution of 14 positional isomers in 7-minutes

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Accucore HPLC Columns – Ruggedness

Efficiency (o-Xylene)

Asymmetry (o-Xylene)

Accucore RP-MS 2.6µm 100x 2.1mm IDMobile Phase: 60/40 ACN/H2OFlow Rate: 400 µL/minInjection Volume: 1 µLColumn Temp: 30°C

Accucore RP-MS 2.6µm 100x 2.1mm IDMobile Phase A: Water (0.05% TFA)Mobile Phase B: Acetonitrile (0.05% TFA)Wash: H2O (0.05% TFA)Injection Volume: 1 µLColumn Temperature: 30°C

4,000 + isocratic test injections with no decrease in performance

6,000 + fast gradient injections with no change in retention

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• Ultimate Core Performance to Maximize Your Investment

• Rugged and reproducible solid core particles• Fast separations with superb resolution• Low backpressures

Conclusion

Core Enhanced Technology