©2011 Waters Corporation 2 · 2012. 5. 2. · .00250025 ID inlet PEEK tubing” ID inlet PEEK tubing .0035” ID outlet PEEK tubing 250nL / 183 µm version for 1mm chemistry (10

Extending the ACQUITY UPLC Family…

©2011 Waters Corporation 1


Introducing ACQUITY UPLC IIntroducing ACQUITY UPLC I--ClassClass

The ACQUITY UPLC I-Class System represents

The ACQUITY UPLC I-Class System accomplishes

g Qg Q

System represents System accomplishes Highest throughput without

compromising performance More recent evolution of UPLC

Based on 7 years of engineering Maximum Peak Capacity

Enhances the performance of any

Based on 7 years of engineering innovation

Fueled by customer inputMass Spectrometer

Better data quality Pinnacle of chromatographic

performance


ACQUITY UPLC IACQUITY UPLC I--Class SystemClass SystemWhat is it?What is it?

New cells for TUV and PDA

Full range of ACQUITY UPLC Column chemistries

— 500 nl/10mm

— 250 nl/10 mm (for 1mm ID columns)

Compatible with FLR, ELSD & all MS

Column Manager Flexibility (CH-A or CM-A)

— Single or multiple column

N APH ith ll ID

Sample manager (SM-FTN or SM-FL)

— Less dispersion volume (tubing valve)

— New APH with smaller ID

Binary solvent manager (BSM)

Less dispersion volume (tubing, valve)

— Improved carry over


— 1200 bar

— New flow path with smaller ID

Instrument Contribution to Instrument Contribution to Bandspread/ExtraBandspread/Extra--Column EffectsColumn Effectsp /p /

22det

2,det

2,

2,

2,

2,

2, Fectorectorvpostcolumnvcolumnvprecolumnvinjectorvtotalv

Injectionvolume

+injector

Tubingbetweeninjector

and

Columnvolume

+frits

Tubingbetweencolumn

and

Band-spreadinginside thedetector

Time-basedBand-

spreadingin theinjector

band-spreading

and column

frits and detector

detectorcell+

tubing

in theDetector

(SamplingRate; TimeConstant)

Engineering developments have specifically improved dispersion

— Injector design, injector volume, fittings, flow path, sealing surfaces

— Reduced tubing volumes - higher pressure/extended flow rate range

— Improved flow cell dispersion


System Band System Band Spreading:Spreading:Effect Effect of of Tubing ID and LengthTubing ID and Length

Tubing with a large ID increases the overall system volume and contributes to band spreading

g gg g

Excessive tubing lengths contribute to band spreading Decreasing tubing ID and length increases system

pressure

Smallest ID is best

Flow Direction

0.020"0.0025" 0.040"

Shortest length is best


Extra Colum VolumeExtra Colum VolumeContribution Of Tubing ID Contribution Of Tubing ID

Tubing ID Volume/foot Volume/100 cm

0 009’’ 12 l 42 l

gg

0.009’’ 12 µl 42 µl

0.005’’ 3.5 µl 12.5 µl

0.004’’ 2.5 µl 8.2 µl

0.003’’ 1.4 µl 4.6 µl

How does it translate to chromatographic performances?— Peak capacity is increased (with decreasing ID)Peak capacity is increased (with decreasing ID)

— Peak shape is improved

— Overal chromatographic performances are strongly impacted

— Pressure drop created with tubings is increasedp g

0.003’’ vs 0.004’’: 50% less volume, 3x more pressure

Higher pressure limit is required, just a tool, a technology enabler

Higher pressure is not a goal


Higher pressure is not a goal

The goal is to maintain the reduced elution volume of resolved UPLC peaks

Two NEW Sample ManagersTwo NEW Sample Managersp gp g

Fixed Loop Flow Through Needle

New EverFlow inject valve designNew EverFlow inject valve designto enable higher pressures

ACQUITY UPLC H-Class chassis and robust rotary sample tray/plate mechanism

New low dispersion fittings, shorter sample path for the FL (10 µL)

Compatible with newest Sample Organizer (18 shelves)

New low dispersion fittings, lower dispersion needle seal for the FTN (<18 µL)

Low dispersion 1, 2, 5 & 10-µL loop design

— Conventional 20, 100, and250 µL available

Optional extension coils

System volume <100 µL100

Omeprazole 10ng 0403 MRM of 1 Channel ES+ 346.083 > 198.068 (Omeprazole)

7.06e7

System volume <95 µL

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

Blank1 10ng 0403 MRM of 1 Channel ES+ 346.083 > 198.068 (Omeprazole)

4.42e3

103

0.20

5

0.24

9

0.27

6

0.29

4

08

psi

16200.00

17100.00

18000.00


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

0.1

0.13

6 0.30

8

13500.00

14400.00

15300.00

Minutes0.00 0.15 0.30 0.45

II--Class FTN vs. IClass FTN vs. I--Class FL:Class FL:Comparison SummaryComparison Summaryp yp y

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 %

P ecision <0.3% Full loop <1% 0.2 to 1.9 μL <0.3% Full loop Precision p<1.0% PLUNO

μ<0.5% 2 to 10 μL

p<1.0% PLUNO

Cycle Time < 15 sec(with load ahead)

< 15 sec(with load ahead)

< 15 sec(with load ahead)

User Simplicity 3 injection modesloop changes

Single injection mode

3 injection modesloop changes


*Measured with a complete system – Measurement on module only doesn’t make sense

A1

Slide 9

A1 Note we have chnaged the spec on teh cycle time both SM's are the same and edited seaker notes to reflect this.Author; 05-10-2011

Comparison SummaryComparison Summaryp yp y


*Measured with a complete system – Measurement on module only doesn’t make sense

A2

Slide 10

A2 Note we have chnaged the spec on teh cycle time both SM's are the same and edited seaker notes to reflect this.Author; 05-10-2011

Column OvensColumn Ovens

NEW Column Heater NEW Column Manager

Active solvent pre-heating

Two (2) Columns plumbed right or left only

Active solvent pre-heating

New 0.003”/0.075 mm I.D. tubing

Robust 1200 bar fitting New 0.003”/0.075 mm I.D. tubing

Robust 1200 Bar fitting

For method development or multi-

Robust 1200 bar fitting

Excellent method transfer between ACQUITY UPLC I-Class Systems

CH-30A is compatible where HPLC method supportcolumns must be supported


ACQUITY UPLC IACQUITY UPLC I--Class SystemClass SystemOptical Detection (TUV & PDA)Optical Detection (TUV & PDA)p ( )p ( )

ACQUITY UPLC I-Class, TUV & PDA

500nL flow cell has been optimized for reduced dispersion

Flow cell body is the samey

Inlet and outlet tubing i.d. has been reduced

0025” ID inlet PEEK tubing .0025 ID inlet PEEK tubing

.0035” ID outlet PEEK tubing

250nL / 183 µm version for 1mm chemistry (10 mm pathlength)


ELSD & FLR are compatible with I-Class

Isocratic Separation ComparisonIsocratic Separation Comparison-- 2.1 x 50 mm column 2.1 x 50 mm column

Instrument Method Name: SampleName: 1290 TCA 50 inj 6

0.44

8

0.55

1 0.68

4

93448.00

60.00

UHPLC low volume configuration; 21 L d b d d0.

1.31

0

1.62

8

2.46

0mA

U

0.00

12.00

24.00

36.00 21 μL measured band spreadAverage USP Plates = 6016

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

646

5

AU

0 024

0.036

0.048

0.060ACQUITY UPLC; 12 μL measured band spreadAverage USP Plates = 7163

1.6

2.48

5

0.000

0.012

0.024

Instrument Method Name: TCA isocratic 50_50 0p8 SampleName: tca 50

01 6

0.10

ACQUITY UPLC I Cl

0.41

5 0.5 0

0.63

6

0.89

1

1.26

2

1.56

4

2.37

3

AU

0.02

0.04

0.06

0.08 ACQUITY UPLC I-Class;5 μL band spreadAverage USP Plates = 11,356


0.00

Minutes0.00 0.40 0.80 1.20 1.60 2.00 2.40 2.80

Lower Lower DDispersion Further Enhances ispersion Further Enhances ACQUITY UPLC I Class PerformanceACQUITY UPLC I Class PerformanceQQ

ACQUITY UPLC

ACQUITY UPLC I-Class


Impact of Low Dispersion:Impact of Low Dispersion:Increase in SensitivityIncrease in Sensitivityyy

1.87x105

1.04x105


ACQUITY UPLC I Class Performance ACQUITY UPLC I Class Performance vsvs Competitive UHPLCCompetitive UHPLCpp

1400.00

UHPLC

0.29

4

0.33

9

mA

U

800.00

1000.00

1200.00

0.19

2

0.22

6

0.25

8

0.32

1

0.39

1

m

200.00

400.00

600.00

0.00

Minutes0.140 0.168 0.196 0.224 0.252 0.280 0.308 0.336 0.364 0.392 0.420 0.448 0.476 0.504 0.532

0.29

2

0.33

71.00

1.20

1.40

ACQUITY UPLCI-Class

0.19

5

9

0.25

7

9

0.38

7

AU

0.40

0.60

0.80


0.22

0.31

0.00

0.20

Minutes0.140 0.168 0.196 0.224 0.252 0.280 0.308 0.336 0.364 0.392 0.420 0.448 0.476 0.504 0.532

II--Class/FTN 2,1mm ID Column Class/FTN 2,1mm ID Column vsvs1mm1mm

1mm column w/1 µl Inj. volume

0.0275AU

0.007AU

2,1mm column w/2 µl Inj. volume

0.013AU

0.0035AU

h h ff h i i


1 mm ID chromatography offers the same retention time, same peak capacity but much more sensitivity with even less injection volume!

SMSM--FTN: UV Carryover PerformanceFTN: UV Carryover Performanceyy

Chlorhexidine Dioctyl Phthalate CaffeineWash Solvent1% Formic Acid in MeOH

Wash SolventAcetonitrile

Wash Solvent80/20 Water/MeOH

AU

0.70

1.40

AU

0.70

1.40

AU

0.60

1.20

Concentration0.25mg/mL



0.00

Minutes0.00 0.50 1.00 1.50 2.00

0.00

Minutes0.00 1.00 2.00 3.00 4.00 5.00

0.00

Minutes0.00 0.50 1.00 1.50 2.00 2.50 3.00

AU

0.006

0.012

AU 0.0012

0.0024

AU

0.008

Standard TraceBlank Trace


0.000 0.0000

0 60 1 20 1 80 2 40 3 00

0.000

0.004

0 50 1 00 1 50 2 00



Minutes1.00 2.00 3.00 4.00 5.00

For concentrations within linear range of detector(<2AU): No Quantifiable Carryover

Minutes0.60 1.20 1.80 2.40 3.00

Minutes0.50 1.00 1.50 2.00

ACQUITY UPLC IACQUITY UPLC I--ClassClassSMSM--FTN: UV Carryover PerformanceFTN: UV Carryover Performanceyy

Chlorhexidine Dioctyl Phthalate CaffeineWash Solvent1% Formic Acid in MeOH

Wash SolventAcetonitrile

Wash Solvent80/20 Water/MeOH

AU

1.00

2.00

AU

1.00

2.00Concentration10mg/mL(40x Higher)

Carryover0.00093%

Concentration10mg/mL(20x Higher)

Carryover0.00020%

AU

1.00

2.00Concentration10mg/mL(40x Higher)

Carryover0.00034%

0.00

Minutes0.00 1.00 2.00 3.00 4.00 5.00

0.00

Minutes0.00 0.60 1.20 1.80 2.40 3.00

0.00

Minutes0.00 0.50 1.00 1.50 2.00

AU

0.002

AU

0.005

0.010

AU

0.008



0.000

0.001

0.000

0.000

0.004



To quantify carryover: High concentrations, above detection linear range, are injected.

Minutes0.60 1.20 1.80 2.40 3.00

Minutes1.00 2.00 3.00 4.00 5.00

Minutes0.50 1.00 1.50 2.00

Carryover Performance: Carryover Performance: Vendor A, ACQUITY Vendor A, ACQUITY UPLC, ACQUITY UPLC, ACQUITY UPLC IUPLC I--Class with Class with XevoXevo TQTQ--SSUPLC, ACQUITY UPLC, ACQUITY UPLC IUPLC I Class with Class with XevoXevo TQTQ SS

High Conc. Standard at Top of Linear Range of DetectorMethod Requirements Carryover to be <20% of LLOQ

0.0136%of LLOQ

LLOQ > s/n = 10LLOD > s/n = 3

n/d n/d n/dTherefore, method Requires <0.001% Carryover

0.0020%

High Conc. Standard Above Linear Range of Detector

Carryover

0.00470%

0.00052% 0.00041%0.00120% 0.00087%20s wash on vendor

A and with multiple


pvalve switches!

Transferring Methods ACQUITY UPLC Transferring Methods ACQUITY UPLC to ACQUITY UPLC Ito ACQUITY UPLC I--ClassClass

Important things to remember— Passive versus active heating

QQ

Passive versus active heating

— System volume differences

— System dispersion differences

Binary versus Quaternary gradients— Binary versus Quaternary gradients


Original Barbiturates Separation Original Barbiturates Separation ACQUITY UPLC HACQUITY UPLC H--Class SystemClass SystemACQUITY UPLC HACQUITY UPLC H Class SystemClass System

1 – Allobarbital2 – Phenobarbital3 – Butalbarbital4 – Mephobarbital5 – Amobarbital6 - Secobarbital


UPLC Method on ACQUITY UPLC HUPLC Method on ACQUITY UPLC H--Class SystemClass SystemClass SystemClass System

Sample: Barbiturates Mix @ 20μM

Column: 2.1 x 50mm ACQUITY UPLC BEH C18, 1.7μmColumn: 2.1 x 50mm ACQUITY UPLC BEH C18, 1.7μm

Injection Volume: 1.5μL

Temperature: 45°C

Fl R t 0 64 L/ iFlow Rate: 0.64mL/min

Mobile Phase A: 10mM Ammonium Bicarbonate Buffer pH 10.0

Mobile Phase B: Acetonitrile

Gradient: Please refer to the table below

Detection Wavelength: 245nm

Time (min) %A %BTime (min) %A %B

Initial 90 10

1.70 68 32


1.71 90 10

Gradient Hold Insert for Gradient Hold Insert for ACQUITY UPLC IACQUITY UPLC I--ClassClassACQUITY UPLC IACQUITY UPLC I--ClassClass


System System Volume Difference Volume Difference Not AdjustedNot Adjusted

1 2

0.28

System System Volume Difference Volume Difference Not AdjustedNot Adjusted

ACQUITY UPLC H-Class

3

4

5

6

AU

007

0.14

0.21

0.00

0.07

ACQUITY UPLC I Cl

1

2

3

4

5

6

U

0.21

0.28ACQUITY UPLC I-Class

AU

000

0.07

0.14


0.00

Minutes0.00 0.30 0.60 0.90 1.20 1.50

Adjusted Gradient Table forAdjusted Gradient Table forACQUITY UPLC IACQUITY UPLC I--Class SystemClass System

ACQUITY UPLC H-Class Gradient Table

ACQUITY UPLC IACQUITY UPLC I--Class SystemClass System

BEFORE NOW

Time (min) %A %B

Initial 90 10

1.70 68 32

Q

1.71 90 10

Gradient Hold insert of 262uL at 640uL/min Flow Rate translates to

ACQUITY UPLC I-Class Gradient Table

Flow Rate translates to 0.40 minutes

Time (min) %A %B

Initial 90 10

0.40 90 10

2 10 68 32


2.10 68 32

2.11 90 10

System System Volume Difference Volume Difference AdjustedAdjusted

1 2

0.28

yy jj

ACQUITY UPLC H-Class

3

4

5

6

AU

007

0.14

0.21

0.00

0.07


1 2

3 4

5

6

U

0.21

0.28ACQUITY UPLC I-Class

AU

000

0.07

0.14


0.00

Minutes0.00 0.30 0.60 0.90 1.20 1.50

Average of 6 Replicate InjectionsAverage of 6 Replicate InjectionsAverage of 6 Replicate InjectionsAverage of 6 Replicate Injections

USP Resolution

Barbiturates Mix ACQUITY UPLC H-Class


Peak 1 and 2 5.70 5.77

Peak 2 and 3 10.74 10.53

Peak 3 and 4 13.30 13.33

Peak 4 and 5 5.32 5.32

Peak 5 and 6 11.28 11.34



1.00

QQ

Peaks Resolution

2-3 3.54ACQUITY UPLC

AU

0.00

0.50 3-4 4.56

7-8 2.02

AU

0.50

1.00 Peaks Resolution

2-3 3.27

3-4 5.20

7 8 2 37

ACQUITY UPLC I-Class – Same Conditions

0.00

1.00

7-8 2.37

Peaks Resolution

2 3 3 61ACQUITY UPLC I-Class – Optimized Transfer*

AU

0.00

0.50

2-3 3.61

3-4 4.80

7-8 2.13


Minutes0.00 0.50 1.00 1.50 2.00

*Compensated for System Volume (20µL) and Temperature Differences (8°C)


Important things to remember— Passive versus active heating

QQ

Passive versus active heating

— Small system volume differences

— System dispersion differences


I‐Class


Third party instrument Third party instrument control – MS

March 2012


ABSciexABSciex /Danaher/Danaher

ACQUITY BSMNanoAcquity UPLC H‐Class UPLC I‐Class

ABI

Analyst V1.42

XX

No 2D

Supports ACQUITY UPLC – Driver Pack v1.40 & June 2011 Driver PackSupports nanoACQUITY UPLC/1D – Driver Pack v1.41

Analyst V1.51

XX

No 2D X X

Enabled with June 2011 Driver PackHot fix v1.52 – supports all previously tested drivers

Analyst V1.60

Not Tested Not Tested Not Tested Not TestedWindows 7 version 32 bit – not currently released – pending date


HystarHystar / / BrukerBrukeryy

ACQUITY BSMNanoAcquityUPLC H‐ClassUPLC I‐Class

Bruker

Hystar V3.2 X

XSupports ACQUITY UPLC – Driver Pack v1.40Supports nanoACQUITY UPLC/1D – Driver Pack v1 41

SR2X

No 2DSupports nanoACQUITY UPLC/1D – Driver Pack v1.41

Hystar V3 2 44 XWith plug in Version 1.5 (Planned on Dec 2011)

Hystar V3.2.44 SR2

XX

No 2DX X


ThermoFisherThermoFisher//DionexDionex

ACQUITY BSM NanoAcquity UPLC H‐Class UPLC I‐Class

Thermo

XcaliburV 2.1

XX

No 2D

Supports ACQUITY UPLC – Driver Pack v1.30Supports nanoACQUITY UPLC/1D – Driver Pack v1.31June 2011 SR2 / March 2011 Driver Packs ‐ Not currently tested ‐ in process [Waters ‐Milford]‐ Issues may delay introduction

XcaliburX

XNo

X

ACQUITY UPLC, ACQUITY UPL I‐Class – June 2011 Driver Pack nanoACQUITY UPLC– March 2011 Driver Pack [v1 42] –

V 2.2X

No 2DNo

No FTNMarch 2011 Driver Pack [v1.42]


Documents

©2011 Waters Corporation 2 · 2012. 5. 2. · .00250025 ID inlet PEEK tubing” ID inlet PEEK tubing .0035” ID outlet PEEK tubing 250nL / 183 µm version for 1mm chemistry (10