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
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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??
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