ACQUITY UPLC Technology Overview Instruments & Chemistry · ©2013 Waters Corporation 1 . ACQUITY...

©2013 Waters Corporation 1

ACQUITY UPLC Technology Overview

Instruments & Chemistry

Bàrbara Bagó, Chemistry Operations Manager Rosa Maria Brunet, System Solutions Sales Manager

Barcelona, 28 October 2014


UPLC® Technology Increasing Resolution, Speed and Sensitivity

UPLC

Reliability

Structural Integrity

Accuracy and

Precision

Low Dispersion

High Speed Detection

Column Technology


The Promise of Small Particles van Deemter Curves (< 500 MW)

HPLC

UPLC


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Minutes 0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00 28.00 30.00

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UPLC®

Rs = 2.2

Rs = 1.7

Traditional HPLC vs. UltraPerformance LC®

UPLC® increases speed to get to right answers…. faster

Rs = 2.4 Rs = 1.8

Traditional HPLC

Run Time = 30 min

Run Time = 4 min


Ultra Performance LC Technology

Years of focused research – At Waters (head quarters) – In collaboration with LC scientists in academia and industry

Introduced the first holistically designed UPLC system in 2004 – ACQUITY UPLC was immediately recognized as a scientific success

– More speed, resolution and sensitivity – >20,000 systems worldwide – >300.000 UPLC Columns worldwide

Holistic design: o Binary Solvent Manager o Sample Manager o UPLC detectors (TUV, PDA) o UPLC columns (BEH C18)


ACQUITY UPLC M-Class H-Class H-Class Bio 2D Clinical

I-Class UPC2 HDX PATROL

UPLC Technology Platform Has Expanded: Adopting Across Organization to Achieve Business Goals


ACQUITY UPLC® H-Class/ H-Class BIO System : HPLC Simplicity, UPLC Performance

Quaternary gradient Variable volume injector

– Flow Through needle Embedded method transfer tools Ease of use UPLC powered

H-Class BIO System

Bio-compatible fluidics for analysis of

biological molecules

AutoBlend Plus simplifies methods

development


ACQUITY UPLC I-Class System

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

New pump heads, check valves, seals, tubings, …

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


Waters Column Product History

Styragel®

µBondapak™

DeltaPak®

1964 1979 1973

1992

Symmetry®

Spherisorb®

Atlantis®

Symmetry® 300

XTerra®

XTerraPrep®

1998

Nova-Pak®

ProteinPakTM

Pico-TagTM

SymmetryShield®

1984

1986

2002

1999 1994 2003

Atlantis® HILIC Silica Prep OBD™ Intelligent SpeedTM

BioSuite™ NanoEase™

2004

ACQUITY UPLC® BEH SunFire™ Columns

PrepPak®

1976

1958

Atlantis® T3 ACQUITY UPLC® HSS T3 AccQTagTM Ultra

BEH130 Columns BEH300 Columns

2006

2005

XBridge™

2007

ACQUITY UPLC® HSS C18 and HSS C18 SB

2008

XBridge™ HILIC

2009

ACQUITY UPLC® BEH Amide ACQUITY UPLC® BEH Glycan XBridge Amide XSelect HSS HPLC Columns

2010

ACQUITY UPLC® BEH200 SEC XSelect CSH HPLC columns

ACQUITY CSH Columns Viridis SFC Columns

ProteinPak High Rs IEX

AccQTagTM

2011

ACQUITY UPLC® HSS Cyano & PFP columns XSelectTM HSS Cyano & PFP columns XP 2.5 µm Columns

2012

ACQUITY UPLC® BEH125 SEC ACQUITY UPC2™ Columns

2013

CORTECS™ Columns ACQUITY UPLC® BEH450 SEC ACQUITY APC™ Columns CSH130 Columns


CORTECS 1.6µ/2.7 µm Columns

UPLC Columns featuring 1.6 µm solid-core silica particles Key benefits:

– Highest efficiency UPLC Column (>35% vs fully porous sub-2-µm columns) – Improved performance at similar backpressure – Increased throughput – Highlight the system band spreading volume difference

(best performance on the UPLC I-Class)

HPLC/UHPLC columns featuring 2.7 µm solid-core silica particles Key benefits

– High efficiency at reduced backpressure o Resolution o Speed

– Scalability to/from UPLC

3 chemistries:

– C18+

– C18

– HILIC

Brochure 720004675en Application Notebook 720004739en


Higher Throughput – Sulfa Drugs: Double the Flow Rate

Comparable peak capacities (Pc) in

half the time

Note: the gradient is scaled to account for the change in flow rate

Comparative separations may not be representative in all applications.

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Competitor Fully-porous C18 at 0.5 mL/min 2.1 x 50 mm 1.8 µm Gradient time: 4.2 min Runtime: 6 min

1

2 3

4

5

6

7

1. Sulfathiazole 2. Sulfamerazine 3. Sulfamethazine 4. Sulfamethoxypyridazine 5. Sulfachloropyridazine 6. Sulfamethoxazole 7. Sulfasoxazole

CORTECS UPLC C18+ at 1.0 mL/min 2.1 x 50 mm 1.6 µm Gradient time: 2.1 min Runtime: 3 min

Pc= 131 31 peaks per minute (gradient)

Pc= 136 65 peaks per minute (gradient)


Impact of System Dispersion on CORTECS Column Efficiency

ACQUITY UPLC I-Class 5.5 µL

ACQUITY UPLC H-Class 12 µL

Acetonitrile/ Water (70/30 v/v), 0.4 mL/ min, 30oC, 0.5 µL injection. Peak i.d.: Acetone, Naphthalene, Acenaphthene 2.1 x 50 mm CORTECS C18 Column

USP N: 18,000

USP N: 11,700

53% Increase in CORTECS Column Efficiency on the ACQUITY UPLC I Class System


Widest Selection of UPLC Particle Offering

CORTECS Solid-Core Technology

Higher efficiency and

resolution

Increased throughput at similar resolution

Higher performance

at same backpressure

BEH Technology

Unparalleled pH stability

Mobile phase and

temperature versatility

Seamless scalability to HPLC

HSS Technology

Maximum retention

Particle and ligand selectivity

Seamless scalability

to HPLC

CSH Technology

Exceptional loading capacity

Superior peak shape

for basic analytes

Seamless scalability to HPLC

Waters is the ONLY company that has fully porous and solid-core sub-2-µm particle columns


UPLC Columns Offering

1.6 [UPLC] 2.7 [HPLC]

1.7 [UPLC] 2.5, 3.5, 5 and 10 µm 2.5, 3.5 and 5 µm HSS & CSH

1.7 [UPLC], 1.8 [UPLC]


ACQUITY Advanced Polymer Chromatography (APC) System

Precise solvent

management

Low system dispersion

Compatibility with challenging

solvents

Rigid, solvent-resilient

columns

Versatile column

management

Stable refractive index

detection

Flexible detection

techniques

Wide range of APC

standards


APC benefits at a glance

MV

0

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10

15

20

25

30

35

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µRIU

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2

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20

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100K

10K

1K

100K

10K

1K

µRIU

0.0

0.4

0.8

1.2

1.6

2.0

2.4

2.8

Minutes 3.70 3.80 3.90 4.00 4.10 4.20 4.30 4.40 4.50 4.60 4.70

MV

-1.0

0.0

1.0

2.0

3.0

4.0

Minutes 20.0 20.5 21.0 21.5 22.0 22.5 23.0 23.5 24.0 24.5 25.0 25.5 26.0

1K polystyrene standard

1K polystyrene standard

GPC APC

3 x Styragel 7.8x300mm (4e, 2, 0.5)

3 x APC TMS 4.6x150mm (200,45,45)


Built upon proven UPLC Technology – Quantifiable increase in productivity

Exceptional increase in available selectivity – Solve routine and complex separation

challenges


UPC2 – New Columns

UPC2 Chiral Columns – TREFOIL (2.5 µm, 3.0 and 2.1mm ID) – Amylose (AMY1), Cellulose (CEL1) and Cellulose (CEL2) – MS compatible

Torus 1.7 µm columns for achiral separations

– Torus 2-PIC most versatile – Torus DEA strong bases – Torus DIOL acids – Torus 1-AA neutrals

1.7 µm BEH Particle

High Density Bonding O Si

O

OO

O

O Si

O

OO

OHNH

N

O SiO

OO

OHNH

O SiO

OO

OHN

O Si

O

OO

OHOH

ACQUITY UPC2

Torus 2-PIC

ACQUITY UPC2

Torus DEA

ACQUITY UPC2

Torus DIOL

ACQUITY UPC2

Torus 1-AA


ACQUITY UPLC Detectors

– Absorbance Detection o Tunable ultraviolet/visible (TUV) o Photodiode array (PDA) o Photodiode array extended

wavelength (PDAeλ)

– Fluorescence (FLR) – Evaporative Light Scattering (ELS)

– Refractive Index (RI)

ACQUITY UPLC Detector Product Line

Low Dispersion

High Data Rates

Maximum Signal:Noise

UPLC Performance


ACQUITY QDa Detector

The ACQUITY QDa Detector is a mass detector designed as an element of a chromatographic separations system


Complementary & Compatible

“Information rich” mass spectral data

– Size of a PDA

Targeted for small molecule applications

Complementary to & compatible with PDA

– UPLC & convergence chromatography UPC2

– Rapid, routine screening & profiling

Benefits of mass detection with the ease of use of a PDA

• 30-1250 DA • PRE-OPTIMIZED ES± • 20 HZ FS • 100 HZ SIR • 4 ORDERS • INTUITIVE •


What do they need?


Method Development Peak Tracking

Track peaks, see coelutions, see more peaks

• RISK MANAGEMENT • WORKFLOW STREAMLINING • PRODUCTIVITY •


USP 37-NF 32 through First Supplement August 1, 2014

Variable USP 36-NF 31 USP 37-NF 32 Through first supplement

Isocratic Gradient

Particle Size -50% L/dp Ratio Constant or N: -25 to + 50%

No changes allowed

Column Length ±70%

No changes allowed

Flow Rate F2=F1 (d22/d12) and ±50% F2-F1 x[(dc22 x dp1)/dC12 x dp2)] and ±50% Not applicable

Column ID

Any allowed if linear velocity is constant

Any allowed if linear velocity is constant

No changes allowed

Injection Volume

Any reduction consistent with precision and detection limits;

no increase permitted

Can be adjusted as consistent with precision and detection

limits

Can be adjusted as consistent with precision and

detection limits

Column Temperature ±10% ±10%

±10%

Mobile Phase pH ±0.2 unit ±0.2 unit

±0.2 unit

F=Flow rate; d = internal column diameter; dc = column diameter, dp = particle size


• Changes made within USP <621> allowable adjustments (USP37-NF32 S1, August 2014) • 10X faster run with 94% solvent savings • Re-validation not required

Isocratic USP Method (UPLC) HPLC to UPLC, 5µm 1.7µm column

0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 0.000

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0.030 HPLC XBridge C

8

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0.030 HPLC USP Res: 6.9

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ACQUITY UPLC H-Class Column: ACQUITY BEH C8, 2.1 x 50mm, 1.7µ (L7) L/dp = 29,400 Flow Rate: 0.61 mL/min, pressure 7700 psi Mobile Phase: 35:15:45 ACN:MeOH:Water Injection Volume: 3.5 µL

Alliance 2695 HPLC Column: XBridge C8, 4.6 x 150mm, 5µ (L7) L/dp = 30,000 Flow Rate: 1.0 mL/min , pressure 1400 psi Mobile Phase: 35:15:45 ACN:MeOH:Water Injection Volume: 50 µL

UPLC USP Res: 7.0

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