ACQUITY UPLC I-Class System - Waters Corporation...©2011 Waters Corporation 5 Tubing Size, μL/foot...

©2011 Waters Corporation 1

ACQUITY UPLC I-Class System


Introducing ACQUITY UPLC I-Class

The ACQUITY UPLC I-Class System represents thepinnacle of ultra performance separations technology, built upon seven years of engineering innovations fueled by customer input.

The ACQUITY UPLC I-Class System accomplishes new levels of analytical capabilities by maximizing peak capacity, advancing the impact of chromatographic separations and extending the performance of any MS detector.


ACQUITY UPLC I-Class Attributes- Meaningful Impact

Pinnacle of Chromatographic

performance

I-Class delivers the highest

throughput without compromising performance

I-Class is designed to optimized the

performance of any Mass Spectrometer

Seamless transfer of existing UPLC

methods

Robust, proven performance built on 7 years of user

based design innovation


Instrument Contribution to Bandspread/Extra-Column Effects

Engineering developments have specifically improved dispersion

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

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

— Improved flow cell dispersion

22

det

2

,det

2

,

2

,

2

,

2

,

2

, Fectorectorvpostcolumnvcolumnvprecolumnvinjectorvtotalv

Injectionvolume

+injectorband-

spreading+

frits

Tubingbetweeninjector

and column

Columnvolume

+frits

Tubingbetweencolumn

and detector

Band-spreadinginside thedetector

cell+

tubing+

frits

Time-basedBand-

spreadingin the

Detector(SamplingRate; TimeConstant)


Tubing Size, μL/foot and Usage

H-Class system APH(0.004” ID – 2.5 μL per foot)

Transfer lines from pump, injector, column, detector and outlet of detector if appropriate

(0.009” ID - 12 μL per foot)

Transfer lines and pump plumbing(0.020” ID - 62 μL per foot)

Transfer lines from pump to autosampler(250 μL per foot)

Microbore systems(0.005” ID - 3.5 μL per foot)

I-Class system APH(0.003” ID – 1.3 μL per foot)

0.040” ID

0.020” ID

0.009” ID

0.005” ID

0.004” ID

0.003” ID


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


System Bandspreading:Effect of Ferrule Fittings

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


System Band Spreading:UV Detector Design

Flow cell is major factor

— Swept volume is at least

equal to the cell volume

— Standard HPLC cells diffuse

separation in as little as 2 μL

Tubing and connections μL

— Size, length and ferrule seating

Acquisition parameters can

be electronic ―diffusers‖

— Points per second

— Smoothing of points


Data Acquisition RatesImpact on UV Chromatography Data

Minutes

0.50 0.52 0.54 0.56 0.58 0.60 0.62 0.64 0.66 0.68 0.70 0.72

1 pt/s

2 pt/s

5 pt/s

40 pt/s

20 pt/s

10 pt/s

How many data points is enough?

Simple rule:15-20 points per peak

The RATE the points are collected is determined by how wide the peak is in TIME at the baseline.

If a peak is only 1 second wide, then you need to collect 20 points

in 1 second (20Hz)

Peaks are ~ 1 - 2 seconds wide

No significant improvement going

to higher rate

0.74


Increasing regulatory demands

Quality and Consistency of results

Evolving analytical demands

Why I-Class?

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


Has minimized dispersion to enhance


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


Why I-Class?


Impact of Low dispersion- Sensitivity

1.87x105

1.04x105


Impact of Low dispersion and optimized gradient – more sensitivity





Vendor B UHPLC System – 60mm FC

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

Improved Dispersion with UVNew UV 500 nL/10 mm Pathlength FC

For UV, resolution and sensitivity is improved





Why I-Class?









T im e0.60 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




C om pound nam e: Om eprazo le

C orre la tion coeffic ient: r = 0 .999985, r^2 = 0 .999970

C alib ra tion curve : 134101 * x + -12 .3789

Response type: E xterna l S td , A rea

C urve type: L inear, Orig in: E xclude, W eighting : 1 /x, A xis trans: None

pg on co lum n0 .0 1 .0 2 .0 3 .0 4 .0 5 .0 6 .0 7 .0 8 .0 9 .0 10 .0

Re

sp

on

se

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




Why I-Class









Isocratic separation comparison- 2.1 x 50 mm column.

Instrument Method Name: SampleName: 1290 TCA 50 inj 6

0.4

48

0.5

51

0.6

84

0.9

34

1.3

10

1.6

28

2.4

60m

AU

0.00

12.00

24.00

36.00

48.00

60.00

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

0.4

44

0.5

50

0.6

85

0.9

33

1.3

20

1.6

46

2.4

85

AU

0.000

0.012

0.024

0.036

0.048

0.060

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

0.4

15

0.5

01

0.6

36

0.8

91

1.2

62

1.5

64

2.3

73

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 spreadAverage USP Plates = 7163

A1290 low volume configuration; 21μL measured band spreadAverage USP Plates = 6016

ACQUITY I-Class 2μl loop;5μL band spreadAverage USP Plates = 11,356


Accelerate Ballistic SeparationsACQUITY and ACQUITY I-Class

Instrument Method Name: 30sec_10_95p600mL_90C ACQUITY

0.1

58

0.2

05

0.3

51

0.4

19

0.4

57

0.4

83

0.5

03

AU

0.000

0.022

0.044

0.066

0.088

0.110

Instrument Method Name: 20sec_NO hold 10_95_p904mL_90C

0.0

98

0.1

17

0.1

70

0.2

13

0.2

41 0.2

61

0.2

76

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 capacityat 5σ = 57

Average peak capacityat 5σ = 70

ACQUITY UPLC

ACQUITY UPLC – I CLASS



Impact

Complex separation challenges require LC systems designed

to maximize the benefits of sub 2µm particle columns. The

ACQUITY UPLC I-Class system:

— Maximized detection sensitivity

— Robust binary based methods

— Increased throughput without compromising chromatographic

fidelity maximizing return on investment


ACQUITY UPLC I-Class System-What is it?

Binary Solvent Manager

Sample Manager (2 options)

— Fixed-Loop (FL) Sample Manager

— Flow-Through-Needle (FTN) Sample Manager

Column Management (2 options)

— Single Column Heater

— Dual Column Manager

o Max 2 columns with optional 2D

Technology feature

Detection

— MS

— TUV or PDA only – new lower dispersion flow cells

New ACQUITY UPLC Systems Driver Pack (DP 3)

Existing ACQUITY UPLC Chemistries


NEW Binary Solvent Manager

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


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 toenable 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 and10 µL loop design

— Conventional 20, 100 and250 µL available

Two NEW Sample Managers


ACQUITY I-Class FTN I-Class FL

Dwell Volume 120 µL 100 µL 95 µL

Bandspread 12 µL


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

100

Omeprazole 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

0.1

03

0.1

36

0.2

05

0.2

49

0.2

76

0.2

94

0.3

08p

si

13500.00

14400.00

15300.00

16200.00

17100.00

18000.00

Minutes0.00 0.15 0.30 0.45 0.60 0.75


NEW Column Heater

New 0.003‖ ID tubing

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




UPLC H-Class to UPLC I-ClassMethod Transfer Example

ACQUITY UPLCH-Class

ACQUITY UPLC I-Class

ACQUITY UPLC H-Class


System Volume Not Adjusted

Adjusted System Volume


->Up time


NEW Column Manager

Two (2) Columns plumbed right or left only

New 0.0003‖ ID tubing

Robust 18 K PSI fitting

Excellent Method transfer between I-Class systems

2D Technology supported



->Up time


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

DimensionsMax. 150mm length x 1.0 or 4.6 mm ID

w/ prefilter or guard column in-lineMax. 150mm length x 1.0 or 4.6 mm ID

w/ prefilter or guard column in-line

Temperature Range Ambient +5°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

NoneTWO valves - 18 K PSI, Dual9 port valve 2-column selector

2D Functionality NoneTWO valves - 18 K PSI, Dual

2 position column selector


Naming Convention

Reinforces that each ACQUITY UPLC Family

member has a unique feature set - ―built for

purpose‖ to address specific user needs

The name support the expansion

of the brand...

...introducing ACQUITY UPLC I-Class

— Inlet

— Innovative

— Investigative


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


ACQUITY UPLC Fast Gradients


ACQUITY UPLC I-Class Faster Gradients for LC/MS

0.3

55

0.3

81

0.4

06

0.4

39 0.4

74

0.4

94

0.5

32

psi

10800.0

12000.0

13200.0

14400.0

Minutes0.00 0.16 0.32 0.48 0.64 0.80

Faster with better resolution


ACQUITY UPLC H-ClassQuaternary Gradient


I-Class



ACQUITY UPLC I-Class is a system deigned to meet evolving

analytical demands, by

— Maximized detection sensitivity, especially MS

— Generating robust binary based methods that will comply

with modern method validation conventions

— Increasing productivity without compromising chromatographic

fidelity to maximize return on investment sooner

Clear where I-Class fits, for the user, now and in the future

Date post:	24-Jan-2021
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