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©2011 Waters Corporation 1
ACQUITY UPLC I-Class System
©2011 Waters Corporation 2
Introducing ACQUITY UPLC I-Class
The ACQUITY UPLC I-Class System represents the pinnacle of ultra performance separations technology, built upon seven years of engineering innovations fueled by customer input.
The ACQUITY UPLC I-Class System is the best MS inlet. It does accomplish new levels of analytical capabilities by maximizing peak capacity, advancing the impact of chromatographic separations.
©2011 Waters Corporation 3
ACQUITY UPLC I-Class System What is it?
Binary solvent manager (BSM) — Enhanced performances, 1200bars
— New tubing with smaller ID
Sample manager (SM-FTN or SM-FL) — Less dispersion volume
— Improved carry over
Column ovens — Single or multiple column
— New APH with smaller ID tubing
New cells for TUV and PDA — 500 nl/10mm
— 250 nl/10 mm (for 1mm ID columns)
Best inlet for all MS detectors
Compatible with FLR & ELSD
©2011 Waters Corporation 4
Complex separation challenges require LC systems that are designed to maximize the benefits of sub 2µm particle columns. — Has minimized dispersion to enhance MS and
UV performance — Lowest carryover complementing MS sensitivity
and extending MS linear dynamic range — The system’s low dispersion, faster cycle-time
allows complex separations can be accelerated without compromising chromatographic fidelity
— Seamless method trasfer
— Robust, proven technology
ACQUITY UPLC I Class
©2011 Waters Corporation 5
Complex separation challenges require LC systems that are designed to maximize the benefits of sub 2µm particle columns. — Has minimized dispersion to enhance MS and
UV performance — Lowest carryover complementing MS sensitivity
and extending MS linear dynamic range — The system’s low dispersion, faster cycle-time
allows complex separations can be accelerated without compromising chromatographic fidelity
— Seamless method trasfer
— Robust, proven technology
ACQUITY UPLC I Class
©2011 Waters Corporation 6
0.020" 0.0025" 0.040"
Shortest length is best
Smallest ID is best
Flow Direction
Large ID tubing increases the overall system volume and contributes to bandspread
Elongated tubing lengths contribute to bandspread Decreased tubing ID and length increases system
pressure
System Band Spreading: Effect of Tubing ID and Length
©2011 Waters Corporation 7
Tubing in the pump flow path that experiences high pressure swings can develop pin holes from the stress and leak
New MP35N lines have a smoother interior surface and are more robust to this phenomenon
Better ID control with MP35N material
Binary Solvent Manager Updated Tubing Material
50x 100x
Conventional SS New MP35N
©2011 Waters Corporation 8
System Bandspreading: Effect of Ferrule Fittings
Void volume in fittings causes Broad Peaks, Tailing, Fronting and Splitting Without External Leaks
©2011 Waters Corporation 9
AU
0.000
0.002
Minutes0.00 2.00 4.00 6.00 8.00
mAU
0.00
2.00
Minutes0.00 2.00 4.00 6.00 8.00
ACQUITY UPLC I-Class System Vendor B UHPLC System
Minutes0.40 0.50 0.60 0.70 0.80
Minutes0.40 0.50 0.60 0.70 0.80
Impact Of Improved Dispersion
Resolution and sensitivity are improved
©2011 Waters Corporation 10
Lower Dispersion I-Class versus Competitor Design
AU
0.000
0.026
0.052
0.078
0.104
0.130
Minutes0.00 0.07 0.14 0.21 0.28 0.35 0.42
ACQUITY UPLC I-Class: 5μL extra column band spread
Average Peak Capacity at 5σ: 70
Competitive UHPLC System: 21μL extra column band spread
AU
0.00
18.00
36.00
54.00
72.00
90.00
Minutes0.07 0.14 0.21 0.28 0.35 0.42
Average Peak Capacity at 5σ : 42
©2011 Waters Corporation 11
Impact of Low dispersion - Sensitivity
1.87x105
1.04x105
©2011 Waters Corporation 12
Complex separation challenges require LC systems that are designed to maximize the benefits of sub 2µm particle columns. — Has minimized dispersion to enhance MS and
UV performance — Lowest carryover complementing MS sensitivity
and extending MS linear dynamic range — The system’s low dispersion, faster cycle-time
allows complex separations can be accelerated without compromising chromatographic fidelity
— Seamless method trasfer
— Robust, proven technology
ACQUITY UPLC I Class
©2011 Waters Corporation 13
MP35N offers a more inert surface that avoid adsorptions
Smother internal surface
Updated Tubing Material
50x 100x
Conventional SS New MP35N
©2011 Waters Corporation 14
After injection and cleaning of the valve and sampling mechanism there is a part of the valve fact that is not cleaned (Noted in RED)
By actuating the valve during the run the material is pushed into the sample loop and then ‘injected’ during the end of the gradient
SM-FL: ‘Double-Click’
Injector
3
2 1
6
5 4
BSM
Column
Sample Syringe
Volume Detector
©2011 Waters Corporation 15
Time0.60 0.62 0.64 0.66 0.68 0.70 0.72 0.74 0.76 0.78 0.80 0.82 0.84 0.86 0.88
%
-1
99
0.60 0.62 0.64 0.66 0.68 0.70 0.72 0.74 0.76 0.78 0.80 0.82 0.84 0.86 0.88
%
-1
99
Carryover Performance: I-Class with Xevo TQ-S
>4 orders of magnitude Assurance that the quantitation represents only the presence of
analyte, not carryover
Qality and Consistency of results
©2011 Waters Corporation 16
Compound name: OmeprazoleCorrelation coefficient: r = 0.999985, r^2 = 0.999970Calibration curve: 134101 * x + -12.3789Response type: External Std, AreaCurve type: Linear, Origin: Exclude, Weighting: 1/x, Axis trans: None
pg on column0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0
Resp
onse
0
200000
400000
600000
800000
1000000
1200000
Exploit the linear dynamic range
Ability to analyze disparate levels using the full sensitivity range of the MS
Carryover Performance: I-Class with Xevo TQ-S
©2011 Waters Corporation 17
When usability, flexibility and lowest possible carryover matter most position the FTN Sample Manager
Excellent dispersion Runs ballistic gradients well Longer cycle time
When ultimate UPLC performance or throughput is the user’s paramount concern, position the Fixed Loop Sample Manager
Best for 1.0 mm columns
Great carryover performance
I-Class FTN vs. I-Class FL: Comparison Summary
Time0.56 0.58 0.60 0.62 0.64 0.66 0.68 0.70 0.72 0.74 0.76 0.78 0.80 0.82 0.84 0.86 0.88
%
0
100
0.56 0.58 0.60 0.62 0.64 0.66 0.68 0.70 0.72 0.74 0.76 0.78 0.80 0.82 0.84 0.86 0.88
%
0
100Omeprazole 10ng 0403 MRM of 1 Channel ES+
346.083 > 198.068 (Omeprazole)7.06e7
Blank1 10ng 0403 MRM of 1 Channel ES+ 346.083 > 198.068 (Omeprazole)
4.42e3
0.10
3
0.13
6
0.20
5
0.24
9
0.27
60.
294
0.30
8psi
13500.00
14400.00
15300.00
16200.00
17100.00
18000.00
Minutes0.00 0.15 0.30 0.45 0.60 0.75
©2011 Waters Corporation 18
ACQUITY I-Class FTN I-Class FL
Dwell Volume 120 µL 100 µL 95 µL
Bandspread 12 µL <9 µL <7 µL
Precision <0.3% Full loop
<1.0% PLUNO 20% to 75% of loop
<1% 0.2 to 1.9 μL <0.5% 2 to 10 μL
<1% 0.2 to 1.9 μL <0.5% 2 to 10 μL
PLUNO 20% to 75% of loop
Carryover 0.005 % 0.001 %* 0.002 %
Sample Discrimination Hydrophobic Samples None Hydrophobic Samples
Cycle Time < 15 sec w/Load Ahead <30 sec < 15 sec w/Load Ahead
User Simplicity 3 injection modes, loop changes Single injection mode 3 injection modes, loop changes
I-Class FTN vs. I-Class FL: Comparison Summary
©2011 Waters Corporation 19
Complex separation challenges require LC systems that are designed to maximize the benefits of sub 2µm particle columns. — Has minimized dispersion to enhance MS and UV
performance — Lowest carryover complementing MS sensitivity and
extending MS linear dynamic range — The system’s low dispersion, faster cycle-time
allows complex separations can be accelerated without compromising chromatographic fidelity
— Seamless method trasfer
— Robust, proven technology
ACQUITY UPLC I Class
©2011 Waters Corporation 20
Isocratic separation comparison - 2.1 x 50 mm column.
Instrument Method Name: SampleName: 1290 TCA 50 inj 6
0.44
8
0.55
1 0.68
4
0.93
4
1.31
0
1.62
8
2.46
0mA
U
0.00
12.00
24.00
36.00
48.00
60.00
Instrument Method Name: TCA isocratic 50_50 0p8 SampleName: tca 50
0.44
4
0.55
0
0.68
5
0.93
3
1.32
0
1.64
6
2.48
5
AU
0.000
0.012
0.024
0.036
0.048
0.060
Instrument Method Name: TCA isocratic 50_50 0p8 SampleName: tca 50
0.41
5 0.50
1
0.63
6
0.89
1
1.26
2
1.56
4
2.37
3
AU
0.00
0.02
0.04
0.06
0.08
0.10
Minutes0.00 0.40 0.80 1.20 1.60 2.00 2.40 2.80
ACQUITY 5μl loop; 12μL measured band spread Average USP Plates = 7163
A1290 low volume configuration; 21μL measured band spread Average USP Plates = 6016
ACQUITY I-Class 2μl loop; 5μL band spread Average USP Plates = 11,356
©2011 Waters Corporation 21
Fastest Possible Scenario (1 mm Column) ACQUITY vs ACQUITY IClass
Instrument Method Name: 30sec_10_95p600mL_90C ACQUITY
0.15
8
0.20
5
0.35
1
0.41
9
0.45
7
0.48
3
0.50
3
AU
0.000
0.022
0.044
0.066
0.088
0.110
Instrument Method Name: 20sec_NO hold 10_95_p904mL_90C
0.09
8
0.11
7
0.17
0
0.21
3
0.24
1 0.26
1
0.27
6
AU
0.000
0.022
0.044
0.066
0.088
0.110
Minutes0.00 0.10 0.20 0.30 0.40 0.50 0.60 0.70
Average peak capacity at 5σ = 57
Average peak capacity at 5σ = 70
ACQUITY UPLC
ACQUITY UPLC – I CLASS
Ballistic gradient on 1mm x 50mm BEH C18
Due to the 15k psi limit, with the ACQUITY, the flow rate is 0.6 ml/mn
With the ACQUITY I-Class the flow rate is 0.9 ml/mn
©2011 Waters Corporation 22
Complex separation challenges require LC systems that are designed to maximize the benefits of sub 2µm particle columns. — Has minimized dispersion to enhance MS and UV
performance — Lowest carryover complementing MS sensitivity and
extending MS linear dynamic range — The system’s low dispersion, faster cycle-time
allows complex separations can be accelerated without compromising chromatographic fidelity
— Seamless method trasfer
— Robust, proven technology
ACQUITY UPLC I Class
©2011 Waters Corporation 23
UPLC H-Class to UPLC I-Class Method Transfer Example
H-Class
I-Class
H-Class
System Volume Not Adjusted
Adjusted System Volume
I-Class
©2011 Waters Corporation 24
AU
0.00
0.50
1.00
AU
0.00
0.50
1.00
AU
0.00
0.50
1.00
Minutes0.00 0.50 1.00 1.50 2.00
*Compensated for System Volume (10µL) and Temperature Differences (8°C)
Transferring Methods ACQUITY UPLC to ACQUITY UPLC I-Class
Peaks Resolution
2-3 3.54
3-4 4.56
7-8 2.02
Peaks Resolution
2-3 3.27
3-4 5.20
7-8 2.37
Peaks Resolution
2-3 3.61
3-4 4.80
7-8 2.13
ACQUITY UPLC I-Class – Same Conditions
ACQUITY UPLC
ACQUITY UPLC I-Class – Optimized Transfer*
©2011 Waters Corporation 25
Complex separation challenges require LC systems that are designed to maximize the benefits of sub 2µm particle columns. — Has minimized dispersion to enhance MS and UV
performance — Lowest carryover complementing MS sensitivity and
extending MS linear dynamic range — The system’s low dispersion, faster cycle-time
allows complex separations can be accelerated without compromising chromatographic fidelity
— Seamless method trasfer
— Robust, proven technology!
ACQUITY UPLC I Class
©2011 Waters Corporation 26
©2011 Waters Corporation 27
ACQUITY UPLC I-Class Conclusion
I-Class incorporates key features such as:
— Minimized dispersion
— Lowest carryover
— Faster cycle-time
— Seamless method trasfer
— Robust, proven technology
ACQUITY UPLC I-Class is a system designed to:
— Provide ultimate performance to the Bioanalysts
— Be the best MS inlet
— Achieve the maximum productivity
— Meet the constantly evolving analytical demands
©2011 Waters Corporation 28
APPENDIX
©2011 Waters Corporation 29
ACQUITY UPLC I-Class Key Product Compatibility
ACQUITY UPLC I-Class is fully compatible with: — All Waters’ current high-end MS detectors — Empower 1154, Empower 2154 and Empower 3 — UNIFI version 1.50
ACQUITY UPLC I-Class is compatible with:
— AB/Sciex Analyst version 1.4.8/1.50 with Driver Pack v1.50
— Regulated users cannot move immediately
ACQUITY UPLC I-Class is not compatible with: — Agilent — Dionex — Thermo
o ACQUITY UPLC should continue to be positioned
©2011 Waters Corporation 30
New Binary Solvent Manager Features
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
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
©2011 Waters Corporation 31
I-Class FTN vs. I-Class FL: Comparison Summary
ACQUITY I-Class FTN I-Class FL
Dwell Volume 120 µL 100 µL 95 µL
Bandspread 12 µL <9 µL <7 µL
Carryover 0.005 % 0.001 % 0.002 %
Precision <0.3% Full loop <1.0% PLUNO
<1% 0.2 to 1.9 μL <0.5% 2 to 10 μL
<0.3% Full loop <1.0% PLUNO
Cycle Time < 15 sec (with load ahead) <30 sec < 15 sec
(with load ahead)
User Simplicity 3 injection modes loop changes
Single injection mode
3 injection modes loop changes
Sample organizer available with both configurations — 18 shelves
©2011 Waters Corporation 32
When usability, flexibility and lowest possible carryover matter most position the FTN Sample Manager
Excellent dispersion Runs ballistic gradients well Longer cycle time
When ultimate UPLC performance or throughput is the user’s paramount concern, position the Fixed Loop Sample Manager
Best for 1.0 mm columns
Great carryover performance
I-Class FTN vs. I-Class FL: Comparison Summary
Time0.56 0.58 0.60 0.62 0.64 0.66 0.68 0.70 0.72 0.74 0.76 0.78 0.80 0.82 0.84 0.86 0.88
%
0
100
0.56 0.58 0.60 0.62 0.64 0.66 0.68 0.70 0.72 0.74 0.76 0.78 0.80 0.82 0.84 0.86 0.88
%
0
100Omeprazole 10ng 0403 MRM of 1 Channel ES+
346.083 > 198.068 (Omeprazole)7.06e7
Blank1 10ng 0403 MRM of 1 Channel ES+ 346.083 > 198.068 (Omeprazole)
4.42e3
0.10
3
0.13
6
0.20
5
0.24
9
0.27
60.
294
0.30
8psi
13500.00
14400.00
15300.00
16200.00
17100.00
18000.00
Minutes0.00 0.15 0.30 0.45 0.60 0.75
©2011 Waters Corporation 33
Column Management
Column management options equivalent to H-Class, lower dispersion, higher pressure, optimized fittings
Maximum 2 columns and 1 Column Manager
I-Class CH-A I-Class CM-A
Column Capacity Single column Up to 2 columns
Dimensions Max. 150mm length x 1.0 or 4.6 mm ID w/ prefilter or guard column in-line
Max. 150mm length x 1.0 or 4.6 mm ID w/ prefilter or guard column in-line
Temperature Range 20°C to 90 °C 4 °C to 90 °C
Solvent Conditioning Active Pre-Heating (APH) independent control of each column with Active Pre-Heating (APH)
Access to MS inlet Door slides across Inlet/Outlet configurable Left or Right
Column Fittings 18 K PSI- New design 18 K PSI- New design
Advanced Functionality None Column selection or 2D
High Pressure Valve Configuration None TWO valves - 18 K PSI, Dual 9 port valve
2-column selector
2D Functionality None TWO valves - 18 K PSI, Dual 2 position column selector
©2011 Waters Corporation 34
High Temp Column Heater With New APH
Supports one UPLC column with guard-column
From ambiant +5°C to 90°C
New 0.003” ID tubing (APH)
Robust 18K psi fitting
Excellent method transfer between I-Class systems
CH-30A is compatible where HPLC columns must be supported — H-Class performance only
©2011 Waters Corporation 35
Column Manager With New APH
Two (2) Columns plumbed right or left only
From 4°C to 90°C
New 0.003” ID tubing (APH)
Robust 18 K PSI fitting
Excellent Method transfer between I-Class systems
2D Technology supported (independent control of HP valves)
©2011 Waters Corporation 36
Evaluating Dispersion - Definition of Bandspreading
w
Peak width at 4.4% of the height of the peak is 5-sigma (5 standard deviations)
tv F σσ ⋅=5w
t =σ
©2011 Waters Corporation 37
Instrument Contribution to Bandspread/Extra-Column Effects
Engineering developments have specifically improved dispersion — Injector, injection volume, fittings, flow path, sealing surfaces
— Reduced tubing volumes
— Improved flow cell dispersion
— Higher pressure/extended flow rate range
22det
2,det
2,
2,
2,
2,
2, Fectorectorvpostcolumnvcolumnvprecolumnvinjectorvtotalv ⋅+++++= τσσσσσσ
Injection volume
+ injector band-
spreading +
frits
Tubing between injector
and column
Column volume
+ frits
Tubing between column
and detector
Band- spreading inside the detector
cell +
tubing +
frits
Time-based Band-
spreading in the
Detector (Sampling Rate; Time Constant)
©2011 Waters Corporation 38
ACQUITY UPLC I-Class BSM
High pressure Binary mixing — 4 solvents
New seals, check valves and
intelligent Intake Valves (i2V)
Extended automatic, compressibility compensation
New vent valve for convenience even at higher pressures
New higher pressure mixers — 50 µL default, 100 µL and 380 µL
Two NEW Binary Solvent Manager
©2011 Waters Corporation 39
Fixed Loop Flow Through Needle
New EverFlow inject valve design to enable higher pressures
H-Class chassis and robust rotary sample tray/plate mechanism
Compatible with newest Sample Organizer (18 shelves)
New low dispersion fittings, lower dispersion needle seal for the FTN
Optional, conventional extension loops
New EverFlow inject valve to enable higher pressures
H-Class chassis and robust rotary sample tray/plate mechanism
Compatible with newest Sample Organizer (18 shelves)
New low dispersion fittings, shorter sample path for the FL (10 µL)
— More robust sample transfer
Low dispersion 1, 2, 5 and 10 µL loop design
— Conventional 20, 100 and 250 µL available
Two NEW Sample Managers