Introducing HALO-5

The First High-Performance

5-Micron Packing in the

World

www.biotech.se

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

0.6 µm

~ 5 µm

SEM of ~ 5 µm HALO Fused-core Graphical representation of ~ 5 µm HALO Fused-core

HALO® -5 Fused-Core Particles

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

C18 (octadecyl)

PFP

(pentafluorophenylpropyl)

Available November 2012

- Phenyl-Hexyl

- C8

- ES-CN

Available February 2013

- 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

Mobile Phase Flow Rate, mL/min

0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0

Pre

ssu

re, ba

r

0

100

200

300

400

500

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

•Columns: 150 x 4.6 mm, C18

•Mobile Phase:

50% ACN, 50% Water

•Temperature: 30°C

•Instruments: Agilent 1100 or

Agilent 1200 SL

0 100 200 300 400

1.8 µm totally porous

3 µm totally porous

2.7 µm HALO fused-core

5 µm totally porous

4 µm HALO fused-core

5 µm HALO fused-core

Plates/Pressure in bar

• When operated at optimum flow rates, HALO-5 columns provide the

highest number of theoretical plates per unit of pressure

More Theoretical Plates at Lower Pressure

5000

10000

15000

20000

25000

30000

35000

0.01 0.1 1 10 100

Pla

te N

um

be

r

µg of 1-Cl-4-Nitrobenzene

2.7 µm Fused-Core, 0.5 µm shell

5 µm Fused-Core, 0.6 µm shell

5 µm Totally porous

HALO-5 Columns Exhibit the Same Sample Loadability as 2.7-Micron

HALO Columns and are Similar to 5-Micron Totally Porous Columns

Features and Benefits

• HALO-5 Columns Can be Substituted for 5-Micron Totally Porous Columns to Provide Sharper Peaks and Shorter Analysis

Times Under Identical Method Parameters

Easy Transfer from Existing Methods

• HALO-5 Columns Provide the Same Efficiency as 3-Micron

Columns at about Half the Pressure

Obtain 5-Micron Benefits for 3-Micron Methods

0,400

0,600

0,800

1,000

1,200

1,400

1,600

1,800

2,000

0,00 0,50 1,00 1,50 2,00 2,50 3,00 3,50 4,00 4,50 5,00

Re

solu

tio

n

Flow, ml/min

2.7-micron Fused-core

5-micron Fused-core

3-micron TP

5-micron TP

Rs = 1.2

Rs = 1.9

Rs = 1.24

Rs = 0.9

Rs = 1.55

Get 3-micron Resolution at 5-micron Pressure with HALO-5

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

Comparable Selectivity with 2.7-Micron HALO

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.