ACQUITY ARC Changing your system without changing your method AU Tech… · Changing your system...

©2016 Waters Corporation 1

ACQUITY ARC

Changing your system without changing

your method

Rainer Rozenich

Separations - Business Development Manager

Eastern European Region


About the ACQUITY ARCTM System

ARC – it is the latest model in the Waters LC product portfolio - designed for HPLC & UHPLC analytics.

Developed for • Routine HPLC analysis • Routine UHPLC analysis • Easy transfers of existing HPLC methods • Method development • Method optimization


Method transfer issues

AU

0.000

0.012

0.024

0.036

0.048

AU

0.000

0.012

0.024

0.036

0.048

Minutes

1.75 2.00 2.25 2.50 2.75 3.00 3.25 3.50 3.75 4.00

„I cannot use the new HPLC, because I do not have the same plot, so I obviously not have equal separation, I see too much or not enough“ „My chromatograms were part of submission, I cannot accept the other instrument“


Understanding Issue Root Cause

• Different system volumes to the column head

-different mixing of mobile phase -different delivery of the gradient

Instrumentation System Volume Dispersion Volume

HPLC ~ 1000µL and above >30µL

UPLC ~ 325 µL or less <12µL

AU

0.00

0.20

0.40

0.60

0.80

1.00

Minutes

0.50 1.00 1.50 2.00

AU

0.00

0.20

0.40

0.60

0.80

1.00

Minutes

0.50 1.00 1.50 2.00 2.50

• Different dispersion volumes

- Flow Cell - Capillaries


European Pharmacopoeia 8.8

Chapter 2.2.9 Liquid Chromatography

„Unless otherwise specified all information below is valid for standard LC as well as for LC using reduced particle-size columns (e.g. sub-2 µm). The latter requires instrumentation characterised by the capability to apply higher pressures (typically up to 100 MPa, i.e. about 15 000 psi), lower extra-column band broadening, improved gradient mixing and a higher sampling rate in the detection system.”

Chapter 2.2.46 Chromatographic Separation Techniques

“Dwell volume. The configuration of the equipment employed may significantly alter the resolution, retention time and relative retentions described. Should this occur, it may be due to excessive dwell volume.” “Monographs preferably include an isocratic step before the start of the gradient programme so that an adaptation can be made to the gradient time points to take account of differences in dwell volume between the system used for method development and that actually used. It is the user’s responsibility to adapt the length of the isocratic step to the analytical equipment used.”


ArcTM UHPLC – Select your volume

Arc Multi-Flow PathTM Technology


ArcTM UHPLC – How it is done

Path 1: 1100µL Path 2: 700µL

No impact on gradient table !



Fine adjustmensts can be done - using Gradient start smart

No impact on gradient table !



AU

0.00

0.02

0.04

AU

0.00

0.02

0.04

AU

0.00

0.02

0.04

Minutes

5.50 6.00 6.50 7.00 7.50 8.00 8.50 9.00 9.50 10.00 10.50 11.00 11.50 12.00 12.50

1) Volume difference to path 1 or 2 can be compensated. 2) Gradient start smart aids to exactly adjust retention time.

A competitor Binary LC System

Arc Path 1

Arc Path 2


European Pharmacopoeia 8.8

Chapter 2.2.46 Chromatographic Separation Techniques Column: replace the chromatographic column by an appropriate capillary tubing (e.g. 1 m × 0.12 mm), use as mobile phase A water, use as mobile phase B: 0.1 per cent V/V solution of acetone Flow rate: set to obtain sufficient back-pressure (e.g. 2 mL/min). Detection: spectrophotometer at 265 nm Determine the time (t0.5) in minutes when the absorbance has increased by 50 per cent (Figure 2.2.46.-4)

Time [min] % A %B

0-20 100->0 0->100

20-30 0 100



If you do not know the system volume, you can determine it

Dwell volume creates an offset before the solvent composition change reaches the inlet of column.


ArcTM UHPLC – Temperature control

CH30-A with new 0.005” APH 30-cm CH and CHC with new low dispersion passive preheater


ArcTM UHPLC – Column Heating

AU

0.00

0.05

0.10

0.15

0.20

Minutes

8.50 9.00 9.50 10.00 10.50 11.00 11.50 12.00

AU

0.00

0.05

0.10

0.15

0.20

Minutes

8.50 9.00 9.50 10.00 10.50 11.00 11.50 12.00

AU

0.00

0.05

0.10

0.15

0.20

Minutes

8.50 9.00 9.50 10.00 10.50 11.00 11.50 12.00

Preheater Preheater

No Preheater No Preheater

AU

0.00

0.05

0.10

0.15

0.20

Minutes

8.50 9.00 9.50 10.00 10.50 11.00 11.50 12.00

Use the suitable heating mode to avoid unwanted impact during transfer

Different system AQCUITY Arc

Different system AQCUITY Arc


Detector Options (LC)

2998 PDA New Low dispersion analytical flow cell

2489 UV/Vis New Low dispersion analytical flow cell

2414 RI

2475 FLR New Low dispersion analytical flow cell

2424 ELS

2432 CD

In addition very special LC detectors such as the electrochemical (2465) detector are available. Also 3rd party detectors can be connected to Empower using the Waters A/D interface (eSATIN).


Dectector Options (MS)

ACQUITY QDa

TQD

TOF

MS detection systems are in multiple configurations available. Single quadrupoles, tandem quadrupoles, time-of-flight, etc. For screening solutions, for characterization, for quantitation, for resarch as well as routine analysis under GXP.


Acquity QDa Detection

Automated Calibration & Pre-optimized ES

Zero Tuning & Disposable Sample Cone

50-1250 Da Range, 10,000 Da/s

+/- Switching, 4 Orders Dynamic Range

PLUG &

PLAY


Acquity QDa Detection

pH3.1, Col3, ACN

Time1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60

AU

0.0

5.0e-2

1.0e-1

1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60

AU

0.0

5.0e-2

1.0e-1

1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60

AU

0.0

2.0e-2

4.0e-2

6.0e-2

8.0e-2

pH3.1, Col3, ACN

m/z400 402 404 406 408 410 412 414 416 418 420 422 424 426 428 430 432 434 436 438 440

%

0

100427.0

429.0

428.0

429.9

pH3.1, Col3, ACN

m/z400 402 404 406 408 410 412 414 416 418 420 422 424 426 428 430 432 434 436 438 440

%

0

100413.0

415.0

pH5, Col3, ACN

m/z400 402 404 406 408 410 412 414 416 418 420 422 424 426 428 430 432 434 436 438 440

%

0

100413.0

415.0

416.0

pH5, Col3, ACN

m/z400 402 404 406 408 410 412 414 416 418 420 422 424 426 428 430 432 434 436 438 440

%

0

100427.0

429.0

430.0

pH4, Col3, ACN

m/z400 402 404 406 408 410 412 414 416 418 420 422 424 426 428 430 432 434 436 438 440

%

0

100413.0

415.0

427.0

416.0 429.0

413.1427.1

413.1

413.1427.1

427.1

427.1

427.1

427.1

413.1

413.1

413.1

pH 3.1

pH 4.0

pH 5.0

m/zm/z

m/z

m/zm/z


Acquity QDa Detection – Easy to use!


About Auto Blend Plus

Program methods directly in units of pH and molarity or in

units of pH and % of organic solvent

Calculation of required proportions from physical constants

– pK, using Henderson-Hasselbalch equation

– “Corrected” pK with salt compensation

– Empirical calibration table covering operating range

Proportions calculated at each pump stroke for best fidelity

Independent gradients for pH and salt concentration

Water Acetonitrile

Alcohol Concentrated

Modifier

AU

0.00

0.05

0.10

AU

0.00

0.06

0.12

AU

0.00

0.05

0.10

AU

0.00

0.05

0.10

AU

0.00

0.05

0.10

Minutes

0.00 5.00 10.00 15.00

pH 6.1

pH 7.6

pH 7.1

pH 7.0

pH 6.9

A

A

A+B

A

A

B

B

B

B

C

C

C

C

C


Auto Blend Plus


Auto Blend Plus

6.35

6.45

6.55

6.65

6.75

6.85

6.95

7.05

7.15

1 2 3 4 5 6 7 8 9

Measured

pH

Fraction Number

Auto•Blend™ Plus Calibration OptionsDelivered pH

Programmed

pK

Corrected pK

Empirical


Benefits using Arc

• Two in One development • Develope new methods using UHPLC mode • Check method in HPLC and UHPLC modes • Cross validate method with both settings (UHPLC/HPLC)

• AutoBlend Plus – let the instrument mix the mobile phase

• Use of stock solutions and pure solvents • Let the instrument mix mobile phase gradient/salt compositions

and let it adjust the pH values • Test multiple elution systems wihtout preparing each mobile phase

manually

System is easy to deploy and implement in the QC/AD labs because you do not need to change your existing method(s) (e.g. gradient table) to obtain the same chromatographic profile(s)/result(s).


Summary

Replicate existing HPLC method

– Regulated EP & USP methods

– Easy transfer from existing platforms

Improve existing HPLC method

– Enable small particles (2.x µm) technology

– Modernization of LC separations

– Easy transfer and method development tools

Accept efficient UPLC methods and transfer to HPLC and (U)HPLC methods

ACQUITY ARC Changing your system without changing your method AU Tech… · Changing your system...

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