ACQUITY UPLC Technology Overview Instruments & Chemistry · ©2013 Waters Corporation 1 . ACQUITY UPLC Technology Overview Instruments & Chemistry . Bàrbara Bagó, Chemistry Operations
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©2013 Waters Corporation 1
ACQUITY UPLC Technology Overview
Instruments & Chemistry
Bàrbara Bagó, Chemistry Operations Manager Rosa Maria Brunet, System Solutions Sales Manager
Barcelona, 28 October 2014
©2013 Waters Corporation 2
UPLC® Technology Increasing Resolution, Speed and Sensitivity
UPLC
Reliability
Structural Integrity
Accuracy and
Precision
Low Dispersion
High Speed Detection
Column Technology
©2013 Waters Corporation 3
The Promise of Small Particles van Deemter Curves (< 500 MW)
HPLC
UPLC
©2013 Waters Corporation 4
AU
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UPLC®
Rs = 2.2
Rs = 1.7
Traditional HPLC vs. UltraPerformance LC®
UPLC® increases speed to get to right answers…. faster
Rs = 2.4 Rs = 1.8
Traditional HPLC
Run Time = 30 min
Run Time = 4 min
©2013 Waters Corporation 5
Ultra Performance LC Technology
Years of focused research – At Waters (head quarters) – In collaboration with LC scientists in academia and industry
Introduced the first holistically designed UPLC system in 2004 – ACQUITY UPLC was immediately recognized as a scientific success
– More speed, resolution and sensitivity – >20,000 systems worldwide – >300.000 UPLC Columns worldwide
Holistic design: o Binary Solvent Manager o Sample Manager o UPLC detectors (TUV, PDA) o UPLC columns (BEH C18)
©2013 Waters Corporation 6
ACQUITY UPLC M-Class H-Class H-Class Bio 2D Clinical
I-Class UPC2 HDX PATROL
UPLC Technology Platform Has Expanded: Adopting Across Organization to Achieve Business Goals
©2013 Waters Corporation 7
ACQUITY UPLC® H-Class/ H-Class BIO System : HPLC Simplicity, UPLC Performance
Quaternary gradient Variable volume injector
– Flow Through needle Embedded method transfer tools Ease of use UPLC powered
H-Class BIO System
Bio-compatible fluidics for analysis of
biological molecules
AutoBlend Plus simplifies methods
development
©2013 Waters Corporation 8
ACQUITY UPLC I-Class System
Binary Solvent Blending – Also known as high pressure mixing, multi-
pump design – Much less system volume than low pressure
mixing design – Can manage very short (< 1min) ballistic
gradients
New pump heads, check valves, seals, tubings, …
Four solvent capability – Solvents A1 or A2 and B1 or B2
Flow rate range – From 0.01 ml/mn to 2 ml/mn
Total system volume – Less than 95 µl (FL) or 100 µl (FTN) including
a 50 µl mixer
Optional 100µl and 425µl mixers
©2013 Waters Corporation 9
Waters Column Product History
Styragel®
µBondapak™
DeltaPak®
1964 1979 1973
1992
Symmetry®
Spherisorb®
Atlantis®
Symmetry® 300
XTerra®
XTerraPrep®
1998
Nova-Pak®
ProteinPakTM
Pico-TagTM
SymmetryShield®
1984
1986
2002
1999 1994 2003
Atlantis® HILIC Silica Prep OBD™ Intelligent SpeedTM
BioSuite™ NanoEase™
2004
ACQUITY UPLC® BEH SunFire™ Columns
PrepPak®
1976
1958
Atlantis® T3 ACQUITY UPLC® HSS T3 AccQTagTM Ultra
BEH130 Columns BEH300 Columns
2006
2005
XBridge™
2007
ACQUITY UPLC® HSS C18 and HSS C18 SB
2008
XBridge™ HILIC
2009
ACQUITY UPLC® BEH Amide ACQUITY UPLC® BEH Glycan XBridge Amide XSelect HSS HPLC Columns
2010
ACQUITY UPLC® BEH200 SEC XSelect CSH HPLC columns
ACQUITY CSH Columns Viridis SFC Columns
ProteinPak High Rs IEX
AccQTagTM
2011
ACQUITY UPLC® HSS Cyano & PFP columns XSelectTM HSS Cyano & PFP columns XP 2.5 µm Columns
2012
ACQUITY UPLC® BEH125 SEC ACQUITY UPC2™ Columns
2013
CORTECS™ Columns ACQUITY UPLC® BEH450 SEC ACQUITY APC™ Columns CSH130 Columns
©2013 Waters Corporation 10
CORTECS 1.6µ/2.7 µm Columns
UPLC Columns featuring 1.6 µm solid-core silica particles Key benefits:
– Highest efficiency UPLC Column (>35% vs fully porous sub-2-µm columns) – Improved performance at similar backpressure – Increased throughput – Highlight the system band spreading volume difference
(best performance on the UPLC I-Class)
HPLC/UHPLC columns featuring 2.7 µm solid-core silica particles Key benefits
– High efficiency at reduced backpressure o Resolution o Speed
– Scalability to/from UPLC
3 chemistries:
– C18+
– C18
– HILIC
Brochure 720004675en Application Notebook 720004739en
©2013 Waters Corporation 11
Higher Throughput – Sulfa Drugs: Double the Flow Rate
Comparable peak capacities (Pc) in
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.
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Competitor Fully-porous C18 at 0.5 mL/min 2.1 x 50 mm 1.8 µm Gradient time: 4.2 min Runtime: 6 min
1
2 3
4
5
6
7
1. Sulfathiazole 2. Sulfamerazine 3. Sulfamethazine 4. Sulfamethoxypyridazine 5. Sulfachloropyridazine 6. Sulfamethoxazole 7. Sulfasoxazole
CORTECS UPLC C18+ at 1.0 mL/min 2.1 x 50 mm 1.6 µm Gradient time: 2.1 min Runtime: 3 min
Pc= 131 31 peaks per minute (gradient)
Pc= 136 65 peaks per minute (gradient)
©2013 Waters Corporation 12
Impact of System Dispersion on CORTECS Column Efficiency
ACQUITY UPLC I-Class 5.5 µL
ACQUITY UPLC H-Class 12 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: 18,000
USP N: 11,700
53% Increase in CORTECS Column Efficiency on the ACQUITY UPLC I Class System
©2013 Waters Corporation 13
Widest Selection of UPLC Particle Offering
CORTECS Solid-Core Technology
Higher efficiency and
resolution
Increased throughput at similar resolution
Higher performance
at same backpressure
BEH Technology
Unparalleled pH stability
Mobile phase and
temperature versatility
Seamless scalability to HPLC
HSS Technology
Maximum retention
Particle and ligand selectivity
Seamless scalability
to HPLC
CSH Technology
Exceptional loading capacity
Superior peak shape
for basic analytes
Seamless scalability to HPLC
Waters is the ONLY company that has fully porous and solid-core sub-2-µm particle columns
©2013 Waters Corporation 14
UPLC Columns Offering
1.6 [UPLC] 2.7 [HPLC]
1.7 [UPLC] 2.5, 3.5, 5 and 10 µm 2.5, 3.5 and 5 µm HSS & CSH
1.7 [UPLC], 1.8 [UPLC]
©2013 Waters Corporation 15
ACQUITY Advanced Polymer Chromatography (APC) System
Precise solvent
management
Low system dispersion
Compatibility with challenging
solvents
Rigid, solvent-resilient
columns
Versatile column
management
Stable refractive index
detection
Flexible detection
techniques
Wide range of APC
standards
©2013 Waters Corporation 16
APC benefits at a glance
MV
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µRIU
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100K
10K
1K
100K
10K
1K
µRIU
0.0
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1.2
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1K polystyrene standard
1K polystyrene standard
GPC APC
3 x Styragel 7.8x300mm (4e, 2, 0.5)
3 x APC TMS 4.6x150mm (200,45,45)
©2013 Waters Corporation 17
Built upon proven UPLC Technology – Quantifiable increase in productivity
Exceptional increase in available selectivity – Solve routine and complex separation
challenges
©2013 Waters Corporation 18
UPC2 – New Columns
UPC2 Chiral Columns – TREFOIL (2.5 µm, 3.0 and 2.1mm ID) – Amylose (AMY1), Cellulose (CEL1) and Cellulose (CEL2) – MS compatible
Torus 1.7 µm columns for achiral separations
– Torus 2-PIC most versatile – Torus DEA strong bases – Torus DIOL acids – Torus 1-AA neutrals
1.7 µm BEH Particle
High Density Bonding O Si
O
OO
O
O Si
O
OO
OHNH
N
O SiO
OO
OHNH
O SiO
OO
OHN
O Si
O
OO
OHOH
ACQUITY UPC2
Torus 2-PIC
ACQUITY UPC2
Torus DEA
ACQUITY UPC2
Torus DIOL
ACQUITY UPC2
Torus 1-AA
©2013 Waters Corporation 19
ACQUITY UPLC Detectors
– Absorbance Detection o Tunable ultraviolet/visible (TUV) o Photodiode array (PDA) o Photodiode array extended
wavelength (PDAeλ)
– Fluorescence (FLR) – Evaporative Light Scattering (ELS)
– Refractive Index (RI)
ACQUITY UPLC Detector Product Line
Low Dispersion
High Data Rates
Maximum Signal:Noise
UPLC Performance
©2013 Waters Corporation 20
ACQUITY QDa Detector
The ACQUITY QDa Detector is a mass detector designed as an element of a chromatographic separations system
©2013 Waters Corporation 21
Complementary & Compatible
“Information rich” mass spectral data
– Size of a PDA
Targeted for small molecule applications
Complementary to & compatible with PDA
– UPLC & convergence chromatography UPC2
– Rapid, routine screening & profiling
Benefits of mass detection with the ease of use of a PDA
• 30-1250 DA • PRE-OPTIMIZED ES± • 20 HZ FS • 100 HZ SIR • 4 ORDERS • INTUITIVE •
©2013 Waters Corporation 22
What do they need?
©2013 Waters Corporation 23
Method Development Peak Tracking
Track peaks, see coelutions, see more peaks
• RISK MANAGEMENT • WORKFLOW STREAMLINING • PRODUCTIVITY •
©2013 Waters Corporation 24
USP 37-NF 32 through First Supplement August 1, 2014
Variable USP 36-NF 31 USP 37-NF 32 Through first supplement
Isocratic Gradient
Particle Size -50% L/dp Ratio Constant or N: -25 to + 50%
No changes allowed
Column Length ±70%
No changes allowed
Flow Rate F2=F1 (d22/d12) and ±50% F2-F1 x[(dc22 x dp1)/dC12 x dp2)] and ±50% Not applicable
Column ID
Any allowed if linear velocity is constant
Any allowed if linear velocity is constant
No changes allowed
Injection Volume
Any reduction consistent with precision and detection limits;
no increase permitted
Can be adjusted as consistent with precision and detection
limits
Can be adjusted as consistent with precision and
detection limits
Column Temperature ±10% ±10%
±10%
Mobile Phase pH ±0.2 unit ±0.2 unit
±0.2 unit
F=Flow rate; d = internal column diameter; dc = column diameter, dp = particle size
©2013 Waters Corporation 25
• Changes made within USP <621> allowable adjustments (USP37-NF32 S1, August 2014) • 10X faster run with 94% solvent savings • Re-validation not required
Isocratic USP Method (UPLC) HPLC to UPLC, 5µm 1.7µm column
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ACQUITY UPLC H-Class Column: ACQUITY BEH C8, 2.1 x 50mm, 1.7µ (L7) L/dp = 29,400 Flow Rate: 0.61 mL/min, pressure 7700 psi Mobile Phase: 35:15:45 ACN:MeOH:Water Injection Volume: 3.5 µL
Alliance 2695 HPLC Column: XBridge C8, 4.6 x 150mm, 5µ (L7) L/dp = 30,000 Flow Rate: 1.0 mL/min , pressure 1400 psi Mobile Phase: 35:15:45 ACN:MeOH:Water Injection Volume: 50 µL
UPLC USP Res: 7.0
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©2013 Waters Corporation 26
Waters on-line communities
©2013 Waters Corporation 27
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