HALO-5 Seminar Slides

Introducing HALO-5 The First High-Performance 5-Micron Packing in the World

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Overview  Introduction  Particle Description  Results: • Efficiency • Backpressure • Loadability

 Features and Benefits  Conclusions

Introduction  HALO core-shell particles are unique high-purity silica particles for stable high efficiency packed columns that are capable of very fast separations with modest back pressures  Original HALO particles are 2.7-microns with 90 Å pores  HALO provides the efficiency of sub-2-micron particles at half the pressure  New HALO-5 particles are ~ 5-microns with 90 Å pores  HALO-5 provides the efficiency of 3-micron particles with the pressure of 5-micron particles

HALO® -5 Fused-Core Particles

~ 5 µm

0.6 µm

SEM of ~ 5 µm HALO Fused-core

Graphical representation of ~ 5 µm HALO Fused-core

Specifications for HALO-5 Particles      

~ 5 μm overall diameter 0.6-μm-thick porous outer shell 90 Angstrom pores in porous shell Highly purified Type B silica Surface area ~100 m2/g Stable at pressures of at least 600 bar

HALO-5 Bonded Phases Available November 2012

- Phenyl-Hexyl - C8 C18 (octadecyl)

- ES-CN

Available February 2013 PFP (pentafluorophenylpropyl)

- HILIC -Penta-HILIC

Performance

• As predicted by theory, the smaller the particle, the smaller the plate height (e.g., the sharper the peaks) • The 4 µm and 2.2 µm particles are from R&D batches

• The reduced plate height, h (same plate height data divided by the particle size), provides a measure of packed bed quality. • The larger-particle HALO packings exhibit lower values for h, indicating more homogeneous packed beds. HALO(2.7) h = 1.5, HALO-5 h = 1.2!

Back Pressure Plots for Columns of Different Particle Sizes 600 2.7 m Halo fused-core 3 m totally porous 4 m Halo fused-core 5 m Halo fused-core 5 m totally porous Linear regression

500

Pressure, bar

400

300

200

•Columns: 150 x 4.6 mm, C18 •Mobile Phase: 50% ACN, 50% Water •Temperature: 30°C •Instruments: Agilent 1100 or Agilent 1200 SL

100

0 0.0

0.5

1.0

1.5

2.0

2.5

3.0

3.5

Mobile Phase Flow Rate, mL/min

4.0

4.5

5.0

More Theoretical Plates at Lower Pressure 5 µm HALO fused-core

4 µm HALO fused-core

5 µm totally porous Plates/Pressure in bar

2.7 µm HALO fused-core

3 µm totally porous

1.8 µm totally porous 0

100

200

300

400

• When operated at optimum flow rates, HALO-5 columns provide the highest number of theoretical plates per unit of pressure

HALO-5 Columns Exhibit the Same Sample Loadability as 2.7-Micron HALO Columns and are Similar to 5-Micron Totally Porous Columns 35000

30000

25000

Plate Number

2.7 µm Fused-Core, 0.5 µm shell 20000

5 µm Fused-Core, 0.6 µm shell 5 µm Totally porous

15000

10000

5000 0.01

0.1

1

µg of 1-Cl-4-Nitrobenzene

10

100

Features and Benefits

Easy Transfer from Existing Methods

• HALO-5 Columns Can be Substituted for 5-Micron Totally Porous Columns to Provide Sharper Peaks and Shorter Analysis Times Under Identical Method Parameters

Obtain 5-Micron Benefits for 3-Micron Methods

• HALO-5 Columns Provide the Same Efficiency as 3-Micron Columns at about Half the Pressure

Get 3-micron Resolution at 5-micron Pressure with HALO-5 Rs = 1.9

2,000

2.7-micron Fused-core

1,800

1,600

3-micron TP Rs = 1.55

Resolution

1,400

5-micron Fused-core

1,200

Rs = 1.24

Rs = 1.2

1,000

5-micron TP 0,800

Rs = 0.9

0,600

0,400 0,00

0,50

1,00

1,50

2,00

2,50

Flow, ml/min

3,00

3,50

4,00

4,50

5,00

Comparable Selectivity with 2.7-Micron HALO PEAK IDENTITIES (in order): 1. Uracil 2. Resorcinol 3. Aniline 4. 4-Chloroaniline 5. Acetoacetanilide 6. Dimethylphthalate 7. Cinnamyl alcohol 8. 2,6-Dinitrotoluene 9. Tolbutamide 10. 4-Chloro-3-nitroanisole

• Methods are easily transferred from HALO 2.7-micron columns to HALO-5 Columns with only minor adjustment to mobile phase strength

Who Should Use HALO-5 Columns?  Users who currently use 3- or 5-micron particle columns and want to increase their efficiency and speed  Anyone wanting to transfer a method to a routine laboratory  Users who want more speed or who want more separation power but are owners of pressure-limited HPLC instruments (400 bar max pressure)

Conclusions Regarding HALO-5 Columns  Produce unusual column efficiency; reduced plate heights h of ~1.2 for small molecules making them the first high-performance 5-micron columns in the world.  Efficiency and separation speed rivals that of 3-μm columns, but with about one-half the pressure.  Thin porous outer shell results in superior mass transfer kinetics and better efficiency at high mobile phase velocities, especially for larger molecules.  2-μm porosity frits allow the convenience and ruggedness of columns with 5 μm particles.  Strong particles allow pressures of at least 600 bar.