nuevas columnas cortecs™ partículas núcleo sólido 1.6 µm · nuevas columnas cortecs™...

Nuevas columnas CORTECS™

Partículas núcleo sólido 1.6 µm

©2013 Waters Corporation 1

Partículas núcleo sólido 1.6 µm

Bàrbara Bagó

Cerdanyola del Vallès, Enero 2014

AGENDAAGENDA

� Introducción

� Nuevas Herramientas Acquity UPLC – Columnas CORTECS ™

©2013 Waters Corporation 2

� Aplicaciones

� Resumen

Primary

Manufacturer BonderColumn

Packers Distributors

Bonding

Silica

(Hybrid)

Bonding

Source & control of silica gel can make a big

difference in your chromatography

Chromatography ManufacturingChromatography Manufacturing

©2013 Waters Corporation 3

Waters

Sales* &

Distribution

Column

Packer

Sales &

Distribution

Column

Packer

Sales &

Distribution

Column

Packer

Sales &

Distribution

* Waters: Worldwide Sales & Distribution3

• Manufactures under cGMP

• Registered with FDA as a medical device manufacturer

Developing a Waters ColumnDeveloping a Waters Column

� Hardware

– Materials

– Design

– Optimization

� Sorbents

©2013 Waters Corporation 4

� Sorbents

– Chemistries

– Synthesis

� Packing

– Equipment Considerations

– Optimization

– Product QC and testing

Developing a Waters Column:Developing a Waters Column:Particle Synthesis and BondingParticle Synthesis and Bonding

� Manufacture of a particle through bonding takes between 4-6

months

– Each step of the synthesis process quality controlled

©2012 Waters Corporation 5

Developing a Waters Column:Developing a Waters Column:Final QC and InspectionFinal QC and Inspection

� Each column Passes:

� Efficiency testing using a test mixture

©2012 Waters Corporation 6

Waters Column Product Waters Column Product HistoryHistory

Styragel®

µBondapak™

DeltaPak®

1992

Symmetry®

Spherisorb®

XTerra®

XTerraPrep®

1998

SymmetryShield®

1984 2002

ACQUITY UPLC® BEH

SunFire™ Columns

PrepPak®

1958

2006

XBridge™

ACQUITY UPLC®

HSS C18 and HSS C18 SB

2008

ACQUITY UPLC® BEH Amide

ACQUITY UPLC® BEH Glycan

XBridge Amide

XSelect HSS HPLC Columns

2010

ACQUITY UPLC® HSS

Cyano & PFP columns

XSelectTM HSS Cyano &

PFP columns

XP 2.5 µm Columns

2012

©2012 Waters Corporation 7

1964 19791973

Atlantis®

Symmetry® 300

Nova-Pak®

ProteinPakTM

Pico-TagTM

1986 19991994 2003

Atlantis® HILIC Silica

Prep OBD™

Intelligent SpeedTM

BioSuite™

NanoEase™

20041976

Atlantis® T3

ACQUITY UPLC® HSS T3

AccQTagTM Ultra

2005 2007

XBridge™ HILIC

2009

ACQUITY UPLC® BEH200 SEC

XSelect CSH HPLC columns

ACQUITY CSH Columns

Viridis SFC Columns

ProteinPak High Rs IEX

AccQTagTM

2011

2012

ACQUITY UPLC®

BEH125 SEC

ACQUITY UPC2

CORTECS™ Columns herald an unprecedented level of

efficiency and sensitivity for UPLC columns.

Featuring 1.6 µm silica-based solid-core particles

IntroducingIntroducing

©2013 Waters Corporation 8

Featuring 1.6 µm silica-based solid-core particles

and ultra-low dispersion hardware,

CORTECS UPLC® Columns enable higher peak capacities

and thereby greater resolution and improved throughput

to meet even the most challenging demands.

CORTECS ColumnsCORTECS Columns

� UPLC Columns featuring 1.6 µm solid-core silica particles

� 3 Chemistries:

– C18+

©2013 Waters Corporation 9

– C18

– HILIC

10 Year Evolution of UPLC 10 Year Evolution of UPLC Particle TechnologyParticle Technology

BEH Technology™BEH Technology™ HSS TechnologyHSS Technology CSH™ TechnologyCSH™ Technology

©2013 Waters Corporation 10

BEH Technology™BEH Technology™

First ACQUITY UPLC particle

Wide pH and temperature range

2004

HSS TechnologyHSS Technology

Enhanced retention

Particle and ligand selectivity

2006

CSH™ TechnologyCSH™ Technology

Designed for selectivity

Improved basic peak shape at low pH

2010

10 Year Evolution of UPLC 10 Year Evolution of UPLC Particle OfferingParticle Offering

CORTECS CORTECS SolidSolid--Core Core BEH TechnologyBEH Technology HSS TechnologyHSS Technology CSH TechnologyCSH Technology

©2013 Waters Corporation 11

SolidSolid--Core Core TechnologyTechnology

Increased efficiency and resolution

Higher throughput

2013

BEH TechnologyBEH Technology

First ACQUITY UPLC particle

Wide pH and temperature range

2004

HSS TechnologyHSS Technology

Enhanced retention

Particle and ligand selectivity

2006

CSH TechnologyCSH Technology

Designed for selectivity

Improved basic peak shape at low pH

2010

The CORTECS SolidThe CORTECS Solid--Core ParticleCore Particle

� Only thin outer layer contains the

pores with the chromatographic

surface

� The center core is nonporous

� The outer shell is typically “bumpy”

CORTECSSolid-core

dp= 1.6 µm

Compared to Fully Porous Particles

©2013 Waters Corporation 12

� The outer shell is typically “bumpy”

not pretty

dcore = 1.12 µm

Rho, ρ = 1.12/1.60 = 0.70

ρ = 0 → fully porous particle

ρ = 1 → nonporous particle

ρ = core diameter / particle diameter

Enabling ExpertiseEnabling ExpertiseDifferentiating Advancements in UPLC ColumnsDifferentiating Advancements in UPLC Columns

Bulk SynthesisRugged sub-2-µm solid-core particlesHighest Efficiencies

©2013 Waters Corporation 13

EngineeringUPLC column hardwareLow band broadening

Column PackingUltra-stable packed column bedsProprietary packing processes

SoftwarePaperless tracking of column history witheCordTM technology

Back to Basics :Back to Basics :Potential for Band SpreadingPotential for Band Spreading

©2013 Waters Corporation 14

Band Spreading will occur along the flow path from the Injector (“Sample Band), into, through and out of the column (“Analyte Bands”), and then into the Detector

C18+ C18 HILIC

Chemistry

Intended Use

General purpose, high-efficiency, reversed-phase column. A positively charged surface delivers excellent peak shape for basic

General purpose, high-efficiency, reversed-phase column. Balanced retention of acids, bases and neutrals at low and mid-range

High-efficiency column designed for retention of extremely polar analytes. Offers orthogonal selectivity vs. C18 columns.

CORTECS Chemistry OverviewCORTECS Chemistry Overview

©2013 Waters Corporation 15

peak shape for basic compounds at low pH.

low and mid-range pH.

vs. C18 columns.

Ligand Type Trifunctional C18 Trifunctional C18 None

Surface Charge Modification

+ None None

Endcap Style Proprietary Proprietary None

Carbon Load 5.7% 6.6% None

Ligand Density 2.4 µmol/m2 2.7 µmol/m2 None

pH Range 2 – 8 2 – 8 1 - 5

Temperature LimitsLow pH = 45 °C High pH = 45 °C

Low pH = 45 °C High pH = 45 °C

Low pH = 45 °C High pH = 45 °C

� Controlled surface charge modification first introduced in 2010

with Charged Surface Hybrids (CSH™ Technology*)

Technology behind CORTECS CTechnology behind CORTECS C1818++

©2013 Waters Corporation 16

� Similar approach used for CORTECS C18+

Unbonded CORTECS Particle

Apply Surface Charge

Bond and Endcap

*Patent Pending

Increased Efficiency of Increased Efficiency of CORTECS ColumnsCORTECS Columns

©2013 Waters Corporation 17

Higher Efficiency leads to Sharper Peaks, Higher Efficiency leads to Sharper Peaks, Better Resolution of Local AnestheticsBetter Resolution of Local Anesthetics

AU

0.00

0.05

0.10

0.15

0.201

2

3

4

5

ACQUITY BEH HILIC2.1 x 50mm 1.7 µm

1. Lidocaine 2. Butacaine 3. Tetracaine4. Procaine5. Procainamide

USP Resolution2,3: 1.2

©2013 Waters Corporation 18

0.00

AU

-0.05

0.00

0.05

0.10

0.15

0.20

Minutes

0.00 0.10 0.20 0.30 0.40 0.50 0.60 0.70 0.80 0.90 1.00

CORTECS UPLC HILIC2.1 x 50 mm 1.6 µm

1

2

3

4

5

USP Resolution2,3: 2.2

Higher Throughput Higher Throughput –– Sulfa Drugs:Sulfa Drugs:Double the Flow RateDouble the Flow Rate

Comparable peak capacities (Pc) in half the time

AU

0.00

0.02

0.04

0.06

0.08

0.10

0.12 Competitor Fully-porous C18 at 0.5 mL/min2.1 x 50 mm 1.8 µmGradient time: 4.2 minRuntime: 6 min

1

23

4

5

6

7

1. Sulfathiazole2. Sulfamerazine3. Sulfamethazine4. Sulfamethoxypyridazine5. Sulfachloropyridazine6. Sulfamethoxazole7. Sulfasoxazole Pc= 131

31 peaks per minute (gradient)

©2013 Waters Corporation 19

half the time

Note: the gradient is scaled to account for the change in flow rate

Comparative separations may not be representative in all applications.

0.00

0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00

CORTECS UPLC C18+ at 1.0 mL/min2.1 x 50 mm 1.6 µmGradient time: 2.1 minRuntime: 3 min

Pc= 13665 peaks per minute (gradient)

Impact of System Dispersion on Impact of System Dispersion on CORTECS Column EfficiencyCORTECS Column Efficiency

ACQUITY UPLC I-Class5.5 µL

ACQUITY UPLC H-Class

USP N: 18,000

USP N: 11,700

©2013 Waters Corporation 20

ACQUITY UPLC H-Class12 µL

Acetonitrile/ Water (70/30 v/v), 0.4 mL/ min, 30oC, 0.5 µL injection. Peak i.d.: Acetone, Naphthalene, Acenaphthene

2.1 x 50 mm CORTECS C18 Column

USP N: 11,700

53% Increase in CORTECS Column Efficiency on the ACQUITY UPLC I Class System

ResumenResumen

� CORTECS Column family features 1.6 µm solid-core silica particles

� Enabled by over 10 years of designing, synthesizing and packing

sub-2-µm particles

� Set the bar as the new efficiency standard for UPLC columns

©2013 Waters Corporation 21

� Three unique chemistries for selectivity and exceptional peak shape

� Higher efficiency leads to improved resolution and faster

throughput

� Lowest dispersion systems result in ultimate efficiency and

performance

¿¿PreguntasPreguntas??

©2013 Waters Corporation 22