Waters ACQUITY UPLC Family...2.1 x 50 mm, 1.7 µm 10 min Compatible with HPLC, UHPLC and UPLC systems 1.368 1.641 1.902 0.00 ACQUITY UPLC H-Class L/dp = 29,411 ACQUITY UPLC BEH C 18

Waters ACQUITY UPLC Family

UPLC Users Meetings Denmark

April, 26-27th

Frederic Forini

Waters European Headquarters

©2012 Waters Corporation 1

“Demand for UHPLC systems has continued to outpace demand for other HPLC techniques. UHPLC t f t UHPLC systems are fast becoming the standard tool for research and routine applications ”research and routine applications.

– Strategic Directions International


Industry Trends:Industry Trends:The Market has ChangedThe Market has ChangedThe Market has ChangedThe Market has Changed

Transitioning from HPLC to UPLC Technology

Increasing number of organizations have realized the business and scientific advantages UPLC Technology

Increased availability of UHPLC instruments provides vendor choice

T h l hift h l d i t l t h t b t tili Technology shift has led companies to evaluate how to best utilize their existing HPLC instruments as they continue to invest in, and transition to, newer UPLC systems.

During this transition, a number of challenges have arisen

that need to be addressed


Resolution Power Provides Opportunity Resolution Power Provides Opportunity to Reduce Run Time to Reduce Run Time Without Compromising ResultsWithout Compromising Results

10

m

AU 0.20

0.30

p gp g

min

A

0.00

0.10

Minutes0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00

1.5

miAU0.20

0.30 HPLC UPLC

Number of samples* 13,500 13,500

Total days 250 38

in

0.00

0.10

0.00 0.50 1.00 1.50 2.50 3.00 3.50Minutes

Total days 250 38

Acetonitrile costs ($100/L) $14,175.00 $715.50

Disposal cost* * $28,350.00 $1,431.00

Total cost $42,530.00 $2,147.00

Cost per sample $3.15 $0.16

Savings per sample $2.99Saving per year $40,383.00

*10 min/analysis, 8 hr/day, 250 working/year


** Assume disposal is two times cost of acquiring

Reducing Environmental Impact Reducing Environmental Impact Without Compromising ResultsWithout Compromising Results

10

m

AU 0.20

0.30 XSelect CSH C18 XP4.6 x 75 mm, 2.5 µm

HPLC System

p gp g

min

A

0.00

0.10

Minutes0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00

1.5

m

ACQUITY UPLC CSH C182.1 x 50 mm, 1.7 µm

UPLC System

AU

0.20

0.30

minA

0.00

0.10

0.00 0.50 1.00 1.50 2.50 3.00 3.50Minutes

Traditional HPLC System 10 min run time

Uses 10.5 ml of ACN

Required disposal 31.5 ml of solvent

ACQUITY® UPLC® System 1.5 min run time

Uses 0.53 ml of ACN

Required disposal 1.19 ml of solvent


(H2O/ACN) (H2O/ACN)

Greener result

Operating costs reduced

Evolution of ACQUITY UPLC TechnologyEvolution of ACQUITY UPLC TechnologyEvolution of ACQUITY UPLC TechnologyEvolution of ACQUITY UPLC Technology20042004 20052005 20062006 20072007 20082008 20092009 20102010 20112011 20122012

Instrument ACQUITY UPLC‐BinaryL I j

ACQUITY UPLC‐Sample O i

Column Oven‐4 column 90CH /C l

ACQUITY UPLC‐Active CVL k

UPLC PATROL‐Process A l

ACQUITY UPLC‐Local Console C ll

UPLC H‐Class‐Quaternary Di I j

UPLC I‐Class‐BinaryL I j

Stay tuned!

‐Loop InjectorColumn Oven‐Single 65CDetectors‐TUV and PDA

OrganizerDetectors‐Revised TUV and PDA‐ELSD

‐Heater/Cooler10 – 90C

‐Leak sensors‐Advanced needle designnanoACQUITY‐CapillaryColumn Oven‐Single 90CDetectors

AnalyzerUPLC Open Access‐open bed autosamplerDetectors‐Extended λPDA

Controller ‐Direct Injector‐Met DevH‐Class BIO‐InertnanoACQUITYHDX‐Hydrogen‐Deuterium

‐Loop Injector‐Direct InjectorACQUITY UPSFC‐SFC / NPUPLC PATROL‐Laboratory AnalyzerTRIZAIC

Chemistry

Detectors‐Fluorescence

PDA Deuterium ExchangeUPLC Online SPE

TRIZAIC‐nanofluidicUPLC 2D

Hybrid 1.7 µmBEH C18

Hybrid 1.7 µmBEHShieldRP18BEH C8BEH Phenyl

Hybrid 1.7 µmBEH HILICBEH 300Å C18AccQ‐Tag Ultra

Silica 1.8 µmHSS C18HSS C18 SB

Hybrid 1.7 µmBEH 300Å C4OST C18

Hybrid 1.7 µmBEH AmideBEH Glycan

Hybrid 1.7 µmCSH C18CSH Fluoro‐PhenylCSH Phenyl‐Hexyl

Silica 1.8 µm‐HSS Cyano‐HSS PFP2.5 µm XP

Size Exclusion‐SEC 125Å

y g

Silica 1.8 µmHSS T3

Guard ColumnVanGuard SFC 1.7 µm

2‐ethylpyridineFluoro‐phenylhybrid

µ‐14 chemistriesSize Exclusion‐SEC 200ÅIon Exchange‐High Res Q‐High Res CM‐High Res SP

Stay tuned!

Mass Spec Technology ACQUITY SQD

ACQUITY TQD3rd party MS control

SYNAPT‐Ion Mobility

XEVO TQ MS XEVO QTof

SYNAPT G2

XEVO G2 QTof

XEVO TQ‐S

SYNAPT G2‐SXEVO TQDACQUITY SQD2

Stay tuned!


Application Kits

AccQ‐Tag Ultra Amino Acid

Peptide Separations

Oligonucleotide Aflatoxin

Intact Proteins

AccQ‐Tag Ultra AminoAcid Analysis for H‐Class

UPLC Technology Platform Has Expanded:UPLC Technology Platform Has Expanded:Adopting Across Organization to Achieve Business GoalsAdopting Across Organization to Achieve Business Goalsp g gp g g

I-ClassH-Class H-Class Bio 2D


nanoACQUITY HDXPATROLUPC2

Transfer Methods With Transfer Methods With MinimalMinimal Disruption to Process…Disruption to Process…MinimalMinimal Disruption to Process…Disruption to Process…

Sub 2 micron Method

HPLC Legacy Method

Sub 2 micron Method

… and Maximum

HPLC Legacy Method

Maximum Impact on

ProductivityHPLC Legacy Method


Ease Ease of Method of Method Transfer Between LC PlatformsTransfer Between LC PlatformsEase Ease of Method of Method Transfer Between LC PlatformsTransfer Between LC Platforms

XSelect HSS T34.6 x 50 mm, 2.5 µm XP

Flow rate = 1.9 mL/min

HPLC

in n /D

aidz

Ura

cil

Gly

citin

Gen

istin

Dai

dze

in

Gly

cite

in

Gen

iste

in

(Daidzin)

Pc = 92

Minutes0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00

1.61

Transfer methods between

XSelect HSS T34.6 x 50 mm, 2.5 µm XP

Flow rate = 1.9 mL/min

ACQUITY UPLC H-Class

P 94

between different LC systems

1.63(Daidzin)

Pc = 94


Minutes0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00

Transferring with Constant L/Transferring with Constant L/dpdp

2.265

4.610

6.916

0.05

XBridge C184.6 x 150 mm, 5 µmAlliance 2695 HPLCL/dp = 30,000

40 min

Transferring with Constant L/Transferring with Constant L/dpdp

12 14 160.00

9 1 4

0.05XBridge BEH C184.6 x 75 mm, 2.5 µm XPAlliance 2695 HPLCL/dp = 30 000

40 min

4X f t th d

3.129

3.731

4.324

0.00

0.05

L/dp = 30,000

XBridge BEH C184.6 x 75 mm, 2.5 µm XPACQUITY UPLC H-Class

10 min4X faster methods

on your HPLC

3.106

3.697

4.275

0.00

0.05

QL/dp = 30,000

ACQUITY UPLC BEH C182.1 x 50 mm, 1.7 µm

10 min Compatible with HPLC, UHPLC and UPLC systems

1.368

1.641

1.902

0.00

ACQUITY UPLC H-ClassL/dp = 29,411

ACQUITY UPLC BEH C182 1 x 50 mm 1 7 µm

4.5 min

Transfer to 1.7 µm UPLC for


1.319

1.582

1.842

0.00

0.05 2.1 x 50 mm, 1.7 µmACQUITY UPLC I-ClassL/dp = 29,411

4.5 min

UPLC for 9X faster methods

Fusion to Fusion to ComplyComply withwith QbDQbDPublishedPublished GuidelinesGuidelinesPublishedPublished GuidelinesGuidelines

Quality by Design: A systematic approach to development that begins with predefinedobjectives and emphasizes product and process objectives and emphasizes product and process understanding and process control, based on sound science and quality risk management

ICH Q8(R2) Guidelines


*ICH: International Conference on Harmonisation

ACQUITY UPLC HACQUITY UPLC H--Class Bio AttributesClass Bio AttributesACQUITY UPLC HACQUITY UPLC H Class Bio AttributesClass Bio Attributes

Based on ACQUITY UPLC H-Class True UPLC performance

Based on ACQUITY UPLC H-

Cl

True UPLC performance

Class

Bio-compatible

AutoBlend+simplifiescompatible

fluidics for biological molecules

simplifiesmeth dev


Extending the ACQUITY UPLC Family…


ACQUITY UPLC IACQUITY UPLC I--Class AttributesClass Attributes

Pinnacle of h t

ACQUITY UPLC IACQUITY UPLC I Class AttributesClass Attributes


chromatographic

performanceHT

without compromising

Built on 7 years user-

based design performancedesign innovation

Compatible with existing

Optimizes the with existing

ACQUITY UPLC & H-

Class methods

the performance of any Mass

Spec


ACQUITY UPLC IACQUITY UPLC I--ClassClassThe Pinnacle of Chromatographic The Pinnacle of Chromatographic PerformancePerformance

Highest resolution offered by Sub 2um particles and low

dispersion fluidics

Accelerated complex separations achieved with low system’s dispersion &

faster cycle-time

Best sensitivity reached with lowest carryover

autosamplerp faster cycle-time p

SYNAPT G2 MS

ACQUITY SQD

ACQUITY TQD

XEVO TQ MS

XEVO TQ-S

Xevo TOFSYNAPT G2 MS

SYNAPT G2 HDMS


Optical Detectors:

TUV; PDA, FLR, ELSD

Q

Rugged and Robust Systems Rugged and Robust Systems Designed to Optimize Productivity Designed to Optimize Productivity g p yg p y

ACQUITY UPLC I-Class System ACQUITY UPLC H-Class System

Ultimate Binary Performance Ultimate Quaternary PerformanceUltimate Binary Performance Ultimate Quaternary Performance

Ideal MS Inlet Methods Transfer & Development

Ultra High Throughput Routine Analysis


Binary Solvent Management Multiple Solvent Management

ACQUITY UPLC with 2D TechnologyACQUITY UPLC with 2D TechnologyACQUITY UPLC with 2D TechnologyACQUITY UPLC with 2D Technology

“The ability to perform two y pdimensional UPLC separations using Waters technology has enabled some of our most challenging assays that require ultra-high sensitivity while maintaining

ll t d d excellent ruggedness and instrument up-time.”

— Rand Jenkins, Director of Research


and Development Pharmaceutical Product Development

(PPD)

ACQUITY Systems with 2D TechnologyACQUITY Systems with 2D Technology

Characterize Increase

Q y gyQ y gy


Characterize the most complex samples

Increase selectivity

and sensitivity

Eliminate unwanted

interferences

Mobile phase

flexibility

The first ever ACQUITY UPLC System with 2D Technology that


provides the proven UPLC benefits of Resolution, Sensitivity and Productivity, now for multidimensional applications.

2D Chromatography With 2D Chromatography With UPLCUPLCWhat Is This?What Is This?What Is This?What Is This?

2D chromatography is a generic concept (from a hardware point of view): Ypoint of view):– UPLC systems configured with 2 or more solvent managers

– One or several HP valves might be required

S l i h b i d

AC

QU

ITY

– Several temperature zones might be required

Three types of applications:

3 types of applications

R l 2D LC Online Sample Parallel C l


Real 2D LC Online Sample Preparation Column Regeneration

Technology EnablersTechnology EnablersWhat Is Required?What Is Required?qq

New ACQUITY 2D Software for Flexibility– Multi-pump multi-valveMulti pump, multi valve

– Basic User Interface

MassLynx softwaref f– Specific version for 2D

CM-A Required– Two HP valves

– Different temperature zones

Configurations with:– ACQUITY UPLC– ACQUITY UPLC

– ACQUITY H-Class

– ACQUITY H-Class Bio

CQ C Cl


– ACQUITY UPLC I-Class


3 types of applicationsyp pp

Real 2D LC Online Sample PreparationParallel Column Regenerationp g

Column 2 is being regenerating while column 1 is used for the analysis

Increase sample throughput

More flexibility, Faster ROI,

Faster Decision Making, Less Waiting Time…


Hardware

Requirements

(2 SM, 2x6PHPV)



Real 2D LC Online Sample Preparation Parallel Column Regeneration

Trap & Elute At Column Dilution

Column 2 is being regenerating while

column 1 is used for the analysis

Sample of interest istrapped and

concentrated on the cartridge

Impurities can betrapped on the

cartridge


Increased samplethroughput

Increased robustness



More flexibility

Fast ROI…


Hardware

Requirements

(2 SM, 1x6PHPV)

Hardware

Requirements

(2 SM, 1x6PHPV)

Hardware

Requirements

(2 SM, 2x6PHPV)

Increase Sample Loading and Increase Sample Loading and Sensitivity with Trap and BackSensitivity with Trap and Back--Transfer Configuration Transfer Configuration

100MRM of 6 Channels ES+

327 > 269.91.80e5

2.14

20 uL Injection Clozapine 1 ppb on 1D System

gg

Time0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50

%

0

250ul Injection on 1 dimension

%


327 > 269.94.28e5

2.73

250 uL Injection on 1D System Peak DistortionVolume Overload

0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50

0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50

%

0

%


327 > 269.91.97e6

250 uL Injection on 2D System

©2012 Waters Corporation 270.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50

0

Loading 1th dimension Gradient Elution 2nd dimension



Real 2D LC Online Sample PreparationParallel Column

Regeneration

Heart Cutting Comprehensive 2D Trap & Elute At Column Dilution

g


Sample of interest istrapped and

concentrated on the cartridge

Impurities can betrapped on the cartridge

An additional pump is configured for

“reconditioning” the sample plug in aqueous solvent before injection

onto the column






Might be used to simplifysample preparation More flexibility


Hardware

Requirements

(2 SM, 1x6PHPV)

Hardware

Requirements

(2 SM, 1x6PHPV)

Might be required in combination with heart

cutting

Hardware

Requirements

(2 SM, 2x6PHPV)

Compound X in 100% ACN at 1 ppb Compound X in 100% ACN at 1 ppb with ACQUITY System with 2D with ACQUITY System with 2D TechnologyTechnologygygy


278.1 > 105 (Amytriptyline)4.01e4

2.69

2 66

20 uL Injected onto 1D System

%

0

2.66

0.35 2.58

2.912.94

3.01

100

1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.000

MRM of 6 Channels ES+ 278.1 > 105 (Amytriptyline)

6.44e50.83

250 uL Injected onto 1D System

1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00

%

00.31

3.49

%


278.1 > 105 (Amytriptyline)1.04e6

7.00250 uL Injected onto 2D System with At-Column Dilution

5 % Water line A


Time1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00

0

5 % Water line A95 % Water line B50 uL mixer post injector



Real 2D LC Online Sample PreparationParallel Column

Regeneration

Heart Cutting Comprehensive Trap & Elute At Column Dilution

g


1/Sample passes throughthe first column

2/Portions of the effluent are sent to the second column

1/Sample passes throughthe first column

2/All or Portions of the effluent are sent to the

second column

Sample of interest is trappedand concentrated on the

cartridge

Impurities can be trapped on the cartridge

An additional pump is configured for

“reconditioning” the sample plug in aqueous solvent before injection onto the

column


Increased selectivity Increased peak capacityIncreased sample

throughput




Might be used to simplifysample preparation More flexibility


Hardware

Requirements

(2 SM, 1x6PHPV)

Hardware

Requirements

(2 SM, 1x6PHPV)

Hardware

Requirements

(2 SM, 1x6PHPV)

Hardware

Requirements

(2 SM, 1x6PHPV)

Might be required in combination with heart

cutting

Hardware

Requirements

(2 SM, 2x6PHPV)

Typical LC Separation versus Typical LC Separation versus Separation Using ACQUITY UPLC with Separation Using ACQUITY UPLC with 2D Technology2D Technologygygy

T i l LC S ti ACQUITY UPLC ith 2D T h l

desired peptides(co-eluting)

phospholipids interference

Typical LC Separation ACQUITY UPLC with 2D Technology

desired peptidesp p

no phospholipids


ACQUITY UPCACQUITY UPC22 SystemSystemACQUITY UPCACQUITY UPC SystemSystem


ACQUITY UPCACQUITY UPC22

The technique Fi t SFC

ACQUITY UPCACQUITY UPC


The technique of choice for normal phase

analysis

First SFC system

designed to fully utilize

sub 2u sub 2u chemistries

UPSFC capability with LC

Familiarity

The technique of choice for

chiraltiseparations

Minimi e cost pe anal sis b ed cing sol ent cons mption b


Minimize cost per analysis by reducing solvent consumption by up to 80%

What is a Supercritical Fluid ?What is a Supercritical Fluid ?pp

Gas - like low viscosity confers high diffusivity and limits pressure drop

Liquid - like high density ensures good solvent properties

High diffusion coefficient enhances kinetic performance of SFC


High diffusion coefficient enhances kinetic performance of SFC

Supercritical Fluid ApplicationsSupercritical Fluid Applicationsp ppp pp

Chromatography (SFC) Analysis & Purification Chiral Separation Chiral Separation Normal Phase

Extraction (SFE) Bioactive compounds Nutraceuticals

SupercriticalCO2

Nutraceuticals Spices and aromatic Decaffeination Decontamination or cleaning

Val able molec les f om

SF Particle Design(RESS & SAS) Polymerization Cristallization Impregnation Valuable molecules from

waste

Reaction

Impregnation


Reaction Hydrogenation Hydroformylation Carboxylation

Why COWhy CO22 ??yy 22

CO2 reaches supercritical state at 31.1°C and 73.8 bar – Its physical state can be easily manipulatedp y y p

CO2 is non toxic, non flammable

CO2 is chemically pure, stable and non-polar solvent, also compatible with LC detectors

Carbon Dioxide as a Green Solvent– Recovered from industrial and fermentation plants

d h d f h ld h b d f– Avoids the production of CO2 that would have been generated from disposal of the solvents it replaces.

– Less time and energy are used to evaporate fractions to get to pure analytes


analytes.

What is Supercritical Fluid What is Supercritical Fluid Chromatography (SFC) ?Chromatography (SFC) ?g p y ( )g p y ( )

CO2 is used as the mobile phase in combination with one or more organic solvent– Wide range of solvent strength

– Composition from 0 to 100% of organic modifier (usually methanol)

A chromatographic technique similar to HPLC A chromatographic technique similar to HPLC

Unique Features Compressible mobile phase Compressible mobile phase Cooling device for CO2 pump Pressure Resistant UV Flow Cell Back Pressure Regulator


Fast Separation AnalysisFast Separation Analysis

Van Van DeemterDeemter curvecurve H = f(u)H = f(u)80.0

100.0

p yp y

20 0

40.0

60.0

H (µ

m)

uopt

Generic conditionsGeneric conditions2‐EP, 150 x 4.6mm, 5m. CO2‐MeOH, 3.5mL/minOven temp @ 40 C BPR @ 200bar

Generic conditionsGeneric conditions2‐EP, 150 x 4.6mm, 5m. CO2‐MeOH, 10mL/minOven temp @ 40 C BPR @ 200bar

3 x uopt

Only 25% efficiency loss

0.0

20.0

0.0 2.0 4.0 6.0 8.0 10.0u (mm/s)

UV detection @ 220nmUV detection @ 220nm

BenzodiazepinesBenzodiazepinesI. MidazolamII. FlunitrazepamIII. LormetazepamIV Cl

0.15

0.20SteroidsSteroids

1. Androstenedione2. Mestanolone3. Testosterone4 S l l

1

23 1

0.15

0.20I II

III

IV. ClorazepateV. NitrazepamVI. Oxazepam

AU

0.00

0.05

0.104. Stanozolol

4

0.60 min

24

3

1.90 min

AU

0.00

0.05

0.10

2.75 min

III

IVIII

V VI0.85 min

IVV

VI


Minutes0.00 0.40 0.80 1.20 1.60 2.00 2.40 2.80 3.20 3.60 4.00

Minutes0.00 0.40 0.80 1.20 1.60 2.00 2.40 2.80 3.20 3.60 4.00

Courtesy of A. Grand-Guillaume Perrenoud, D. Guillarme, Pr J-L. Veuthey, University of Geneva

High Resolution AnalysisHigh Resolution Analysis

BZDBZD 150150 x 4 6mm 5x 4 6mm 5 mmAnalytical conditions : Analytical conditions : CO2‐MeOH gradient mode, 4mL/min ; PrincetonSFC 2EP 150 x 4.6mm, 5m; Oven temp @ 40 C ; BPR @ 150bar ; UV @ 220nm

g yg y

BZDBZD1. Diazepam2. Midazolam3. Flunitrazepam

150150 x 4.6mm, 5x 4.6mm, 5mm30bar30bar

213 5 + 6

710 + 11

P 634. Lormetazepam5. Flurazepam6. Alprazolam7. Triazolam8. Clorazepate9 Bromazepam

21

48

912 13 + 14

15

Pc = 63

9. Bromazepam10. Nitrazepam11. Clonazepam12. Oxazepam13. Lorazepam14. Clozapine

450450 x 4.6mm, 5x 4.6mm, 5mm80b80b

p15. Olanzapine

N

NR2

R3R4

R1

213

4

6

78 9 1112

1314

80bar80bar

510

15

Pc = 108


R5

Courtesy of A. Grand-Guillaume Perrenoud, D. Guillarme, Pr J-L. Veuthey, University of Geneva

RememberRemember::Smaller Particles the Enabler of ProductivitySmaller Particles the Enabler of Productivityyy


SFC Column Efficiency CurvesSFC Column Efficiency Curvesyy

5.0 µm XBridge™ HILIC, 3.0x50 mm



µ g ,

1.7 µm ACQUITY BEH , 3.0x50 mm

Isocratic Conditions


CO2/Methanol 85:15 130 bar outlet pressure

Resolution ComparisonResolution Comparisonpp

0 14

0.16

5 µm 3.0x100 mm Column

AU

0.08

0.10

0.12

0.14µ

2 mL/minuteCO2/Methanol Gradient

0.00

0.02

0.04

0.06

0.20

0.25 1.7 µm

AU

0 05

0.10

0.15


0.00

0.05

Minutes0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00

nanoACQUITYnanoACQUITY

Inlet of choice N UPLC

nanoACQUITYnanoACQUITY


Inlet of choice for High

Resolution MS systems

Nano UPLC for 3rd party MS systems

System of choice for sample limited l

Direct flow, non-split system

applications


PATROL Laboratory Analysis SystemPATROL Laboratory Analysis System

Seamless Mi i i

PATROL Laboratory Analysis SystemPATROL Laboratory Analysis System


Seamless transfer of optimized

methods from the Process L b t Mf

Minimize error,

optimize methods,

and improve Lab to Mfg floor

and improve workflow

Expand laboratory capabilities from QC to

Bridges pilot plant to mfg

Process Development

Inc ease th o ghp t fo the Reaction Monito ing/Optimi ation


Increase throughput for the Reaction Monitoring/Optimization Labs

Sample Work Flow ComparisonSample Work Flow ComparisonACQUITY & PATROL AtlineACQUITY & PATROL AtlineQQ

LOADVIAL

ASPIRATESAMPLE

INJECTSAMPLE

ACQUITY SAMPLE

MANAGER

LOADVIAL

VERIFYVIAL

READBARCODE

VALIDATESAMPLE

ASPIRATESAMPLE

DILUTESAMPLE

INJECTSAMPLE

PATROLPROCESSSAMPLE

MANAGER


Product Life CycleProduct Life CycleAnalytical SupportAnalytical SupportAnalytical SupportAnalytical Support

DATA and INFORMATICSSpecific to ManufacturingSpecific to Manufacturing

VibrationalSpectroscopy


Product Life CycleProduct Life CycleAnalytical SupportAnalytical SupportAnalytical SupportAnalytical Support

DATA and INFORMATICS


Hydrogen Deuterium Exchange (HDX)Hydrogen Deuterium Exchange (HDX)What is it?What is it?

Measures changes in protein conformation

Application Space– Drug Binding

– Protein Product Conformation and Stability

– Protein-Protein Interactions


Typical HDX WorkflowTypical HDX Workflowypyp

D OAt various times,

li t D O

Add D2O, pH 7

D2O

D2O

D2O

D2O

D2O

D2OD2O

LabelingEquilibrate

move an aliquotto quench buffer

Quenched

D2O

D2O

D2O

D2O

D2O

D2OD2O

Labeling25 C, pH 7

Equilibrate25 C, pH 7

QuenchedpH 2.5

++ +

Mass

UPLCElectrosprayMass Spectrometry

+ +

Analyzer

Peptides

Online Digestion

0° C


p y

nanoACQUITY UPLC System nanoACQUITY UPLC System with HDX Technologywith HDX Technology

nanoACQUITY UPLC System with HDX Technology

gygy

consisting of:

nanoACQUITY Binary Solvent Manager

Auxiliary Solvent Manager

HDX Manager (replaces sample manager)

nanoACQUITY UPLC System with HDX Automation

consisting of:

ACQUITY Bi S l t MnanoACQUITY Binary Solvent Manager

Auxiliary Solvent ManagerHDX Manager (replaces sample manager)

Leap HDX Automation Manager (not shown)


Leap HDX Automation Manager (not shown)

nanoACQUITY UPLC HDX ManagernanoACQUITY UPLC HDX ManagernanoACQUITY UPLC HDX ManagernanoACQUITY UPLC HDX Manager

Injection PortInjection PortInjection PortInjection Port Injection ValveInjection ValveInjection ValveInjection Valve Trap ValveTrap ValveTrap ValveTrap Valve


Column HeaterColumn Heater(Not Visible)(Not Visible)

Column HeaterColumn Heater(Not Visible)(Not Visible) Analytical ColumnAnalytical ColumnAnalytical ColumnAnalytical Column

Documents

Waters ACQUITY UPLC Family...2.1 x 50 mm, 1.7 µm 10 min Compatible with HPLC, UHPLC and UPLC systems 1.368 1.641 1.902 0.00 ACQUITY UPLC H-Class L/dp = 29,411 ACQUITY UPLC BEH C 18