ACQUITY UPLC I-Class System - Waters Corporation · The ACQUITY UPLC I-Class System represents the...

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ACQUITY UPLC I-Class System


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.


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


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


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


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


System Bandspreading: Effect of Ferrule Fittings

Void volume in fittings causes Broad Peaks, Tailing, Fronting and Splitting Without External Leaks


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


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


Impact of Low dispersion - Sensitivity

1.87x105

1.04x105


MP35N offers a more inert surface that avoid adsorptions

Smother internal surface

Updated Tubing Material

50x 100x

Conventional SS New MP35N


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


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


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


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


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



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






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


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


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


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*







— Robust, proven technology!



ACQUITY UPLC I-Class Conclusion

I-Class incorporates key features such as:

— Minimized dispersion

— Lowest carryover

— Faster cycle-time



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


APPENDIX


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


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



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


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


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


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


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


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)


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


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)


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


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

ACQUITY UPLC I-Class System - Waters Corporation · The ACQUITY UPLC I-Class System represents the...

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