agilent 1290 infinity lc · agilent 1290 infinity lc - separation power per time page 12 june 06,...

June 06, 2005

Agilent 1290 Infinity LC

Dr. Michael KraftPharma Industry Marketing ManagerWaldbronn, Germany

Agenda

• 1290 Infinity LC• UHPLC: Situation & Challenges• Performance Examples• System Overview • 1290 Infinity Pump• 1290 Infinity DAD• 1290 Infinity Autosampler and 1290 Infinity Flexible Cube• 1290 Infinity Thermostatted Column Compartment• Gradient Behaviour and Method Transfer• 1290 Infinity LC for LC/MS• 3rd party SW – Instrument control• Summary

What Debate ?

Why is it Over ?

Short History of UHPLC

2003 Agilent 1200 Binary 1.8um RRHT Columns2004 Waters Acquity 1.7um BEH Columns2006 Agilent 1200 RRLC

Thermo Accela 2007 Shimadzu UFLCxr

Jasco Xtreme-LCHitachi LaChrom UltraScient. Syst. Ultra HP

2008 Dionex RSLCKnauer Platin Blue

2009 PE Flexar

10 New UHPLC Systems in 5 Years – All different.

UHPLC Situation 2009 – Power Ranges

bar

ml/min

Standard LC

Vendor A

Agilent RRLC

2.1mm ID 3 – 4.6mm ID

STM and SP (superficially porous) columnsSTM columns

Vendor E

Vendor C

Vendor D

Power = Pressure x Flow Rate

UHPLC – Situation 2009

Problems

• Method Transfer• Not possible• Requires re-development

and re-validation• Performance Limitations

• Speed • Resolution • Sensitivity • Precision• Linearity

Differencies

• System Volumes

• Pressures

• Flow Rates

• Temperatures

• Data rates

• Sensitivity

• Injection principle

• ...

10 New UHPLC Systems in 5 Years – All different.

STM = Sub-2um SP = Superficially PorousTP = Totally Porous

Debate

• Methods: transferable?

• ID: 4.6, 3 or 2.1 mm?

• Particle: TP or SP?

• Size: STM or larger?

• Flow: high or low?

• T: high or low?

• P: 600 or 1000bar?

UHPLC Situation 2009- An “Ideal” System and Its Benefits - Turning “Or” to “And”

Ideal: A System that solves all problems ... and ends the UHPLC debate.Characteristics

• Maximum Power Range • enclosing any other (U)HPLC

system power range

• Smallest System Volume• tools that allow to emulate any

other (U)HPLC system volume

• Best-in-class Spec/Performance• All categories

• Flexible Injection Principle• Variable loop and fixed loop

Benefits• Seamless Method Transfer

• From / To: ANY (U)HPLC system

• No Exclusion / Perform. Limitation • Column ID: 2.1 AND 3 / 4.6mm ID

• Particle size: STM AND larger

• Particle type: TP AND SP

• Flow rates: low AND high

• Pressures: high at ANY flow rate

• Solvent: ACN AND Methanol

• Detection: UV AND MS

STM = Sub-2um SP = Superficially PorousTP = Totally Porous

June 06, 2005


1290 Infinity

1290 Infinity – Compatible with any LC

bar

ml/min

Standard LC

Vendor A

Agilent RRLCVendor B

Vendor C

Vendor D

A new power range providing maximum performance, flexibility, compatibility and investment protection

AdvantagesSupports all analytical columns

Enables Method Transfer

More Resolution

More Speed

More viscous solvents

Lower temperatures

Agilent 1290 Infinity LC- Separation Power per time

. Tranquilizer SampleColumn: Superficially porous

50 x 2.1mm, 2.7umFlow 3.5ml/min, ~ 700 bar

Gradient 2-92% ACN in 0.4minT= 80 °C

0.095sec PW

0.140secPW

*Pc = based on 2nd peak W(1/2)

Peak capacity* ~ 170 in 24 sec.World record !


NH

NH

O

O

OH

O

OHNH

N Cl

O

O

OH

OH

GlafeninePeak 3

ON

ClH

Diphenhydramine.HClPeak 4

N

ClH

Amitriptyline.HCl Peak 5

O

O

O

O

O

OO

O

(+)-Bibenzyl-L-tatratePeak 7

DiazepamPeak 8

N

NCl

O

NH

OH

O

Cl

Cl

DiclofenacPeak 9

NaphthalenePeak 10

UracilPeak 1

Tropic acidPeak 2

Methyl benzoatePeak 6

10 Compound Pharma Test Mix


June 06, 2005

F = 2 ml/minP = 700 bar

0.6min

H2O (0.05% Formicacid) / 2-98 % ACN T = 80°CV_i = 1 µl WL = 210 nm DR = 80 Hz



C18 2.1 x 50mm, 1.7 µm


Agilent ZORBAX RRHD SB C18 2.1 x 100mm, 1.8 µm

H2O (0.05% Formic acid) / 2-98 % Acetonitrile T = 80°CV_i = 0.5 µl WL = 210 nm DR = 80 Hz


1.0min

Baseline separation of Glafenine diastereomere achieved!Less than 1 minute analysis time!


14

Pharma: Initial HPLC to Final 1290 UHPLC Method(Determination of impurities and related substance of metoclopramide hydrochloride in a pharmaceutical formulation)

- Highest Speed with Highest Sensitivity & Precision

14 min

3 min

150 x 3.0 mm, 3.5 µm

100 x 2.1 mm, 1.7 µmP_max = 880bar

• Straightforward method transfer• 4 x faster on 1290 with same accuracy and resolution• Outstanding sensitivity

Gain N and Rs with RRHD Long Lengths150 +100 mm = 250 mm 1.8um @ 1000 bar

Group/Presentation TitleAgilent Restricted

Month ##, 200X

α3,2: 1.15Rs3,2: 6.75N6 : 49200Pw2: 0.017 minPw6: 0.029 min

α3,2: 1.15Rs3,2: 4.92N6 : 29600Pw2: 0.015 minPw6: 0.023 min

Mobile Phase 20:80 water:acetonitrile, Flow Rate: 0.5 mL/min, ambient temperature, ZORBAX RRHD SB-C18, 1.8um columns

150 mm (P:560bar)

150 +100 mm = 250mm (P: 960 bar)1. Uracil2. Acetophenone3. Propiophenone4. Butyrophenone5. Valerophenone6. Hexanophenone

1

2

6

543

1

min0 0.5 1 1.5 2 2.5 3 3.5

mAU

0

50

100

150

200

250

300

min0 0.5 1 1.5 2 2.5 3 3.5

mAU

0

50

100

150

200

250

300

Highest Rs in less than 3 minutes, 250mm, 1.8um at 1000 bar

Agilent 1290 Infinity LC - Infinite possibilities

1290 Infinity Binary Pump1290 Infinity Binary Pump

June 25, 2009

Agilent Confidential

1290 Infinity Binary PumpNext generation Pump Technology - How it looks like inside

June 25, 2009

Channel A

ChannelB

Jet Weaver

Multi-layerHeat exchanger

Solvent selectionvalveDegasser

PurgeValve

Silicon CarbidePistons

High ResolutionPump drives Next Generation Technology

Highest Speed and Resolution:1200bar @ 2mL/min800bar @ 5mL/min Lowest Delay volume: 10 µl / 45 µlLowest Mixing NoiseHighest RT PrecisionAutomatic PurgeAutomatic Leak Correction

1290 Infinity Binary Pump- How it looks like inside

New piston material:SSiC (Sintered Silicon Carbide)

1) Improved thermal conductivity for higher seal lifetime and instrument uptime

2) Enabling higher pressure at higher flow rates (1200 bar at 2 ml/min)

1290 Infinity LC – Delay Volume and Mixing Noise- How to achieve lowest delay volume and lowest noise?

1200 SeriesSolvent A

Passive Damper0-300 ul

Passive Mixer400 ul

1200 SeriesSolvent B

1290 InifinitySolvent A

1290 InifinitySolvent B

Very Good Noise~ 800 µl

+ =Active Damping

10µlMicrofluidic Mixing

35µl

10µl / 45µl DV:80x / 20x lower

Lowest Noise< 50%

Agilent Jet Weaver- the next generation in high-efficient mixing

Technology:Multi-layer technology - Diffusion bonded stainless steel,etched structure (Proprietary Microfluidics Technology)

*Mixer Volume is preliminary

Combining highest mixing efficiency with lowest delay volume (35 µl and 200 µl for TFA applications) for lowest UV noise.

Precision of Retention times for fast LC- Overlay of 10 runs

Precision Peak 1 Peak 2 Peak 3 Peak 3 Peak 5 Peak 6 Peak 7 Peak 8 Peak 9

% RSD RT 0.016 0.0077 0.0052 0.0058 0.0058 0.0054 0.0059 0.0057 0.0080

% RSD Area 0.5 µl 0.401 0.393 0.429 0.470 0.398 0.463 0.437 0.518 0.555

2.1x50mm Zorbax RRHD SB C18, 1.8µmSample: Phenone Standard

Injection volume: 0.5ulMobile phase: H2O/ACN with Jetweaver

0 min 5%ACN, at 0.5min 95% ACNFlow rate: 1.2ml/min

June 06, 2005

1290 Infinity Diode-Array Detector1290 Infinity Diode-Array Detector

1290 Infinity Diode-Array Detector- The Next Generation in UV-Vis Detection

Deuterium lamp

Max-Light Cartridge Cell

Programmableslit

1024 elementdiode-array

GratingMirror

Next Generation Flow Cell Design

Non-Coated Fused Silica Optofluidic Waveguides

1290 Infinity Diode Array Detector- highest sensitivity under any condition

1290 Infinity DAD

1200 DAD SL

1200 RRLC (no mixer, damper)5-95% ACN, 0.5 ml/min 80C, 40 Hz, 230nm/360nm

Next Level Sensitivity and ProductivityMore automated integration at any condition2 – 20x higher sens with 1 cm Cell8 – 80x higher sens with 6 cm UHS Cell

1290 Infinity Diode Array Detector- Ultra Sensitivity by 6cm Flow Cell

min0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8

mAU

0

5

10

15

20

25

30

35

40

DAD1 B, DAD1A, DAD: Signal A, 251 nm/Bw:10 nm Ref 360 nm/Bw:100 nm (F:\AE4-ANTHRACENE\ANTHRA-STD-CEL7.D)DAD1 B, DAD1A, DAD: Signal A, 251 nm/Bw:10 nm Ref 360 nm/Bw:100 nm (F:\AE4-ANTHRACENE\ANTHRA-HS-CEL07.D)

High-Sensitivity Standard Cell (1cm)S/N: 612

Anthracene, 16.4pg/µlWater/ACN = 30/700.5ml/min, 1µl injection36C, 251nm, PW>0.05min

Ultra High-Sensitivity Cell (6cm)S/N: 2339

4 x Higher Sensitivity

1290 Infinity Autosampler1290 Infinity FlexCube

1290 Infinity Autosampler1290 Infinity FlexCube

1290 Infinity Autosampler- Next Generation Variable Loop Injector

AdvantagesLower volume (70ul)

Highest precision w/o loop change from 0.1 – 40ul

No loss of sample

Lowest Carry Over

<0.002% Chlorhexidine

New Cartridge Loop

1290 Infinity FlexCube- Extending the capabilities

Syringe PumpRail for

external Valve

Valve option 1 Valve option 2Low pressureValve

Applications:a) Automated needle seat back-flushing for lower carry overb) Fixed loop injection mode for fastest cycle timesc) and more ...........

1290 Infinity FlexCube- Automated needle seat backflushing - Fixed loop injection mode

AdvantagesMore robust lowest carry overPrevention of needle seat blockages

AdvantagesLow Delay volume (10 µl)

Overlapped injection with very low carry over

Fixed Loop vs. Flow-through Variable LoopFlow-Through Fixed Loop

Injection Precision for Fixed Volume ■■■■■ ■■■■■Injection Precision for a Wider Range of Volume (w/o hardware change)

■■■■■ ■■■■

Injection precision for small Volumes(< 1 µl)

■■■■■ ■■■

Sample Losses ■■■■■ ■■■■

Carry-over Performance ■■■■ ■■Hydraulic Delay Volume ■ ■■■■■Cycle time reductionthrough overlapped injection

■■ ■■■■■

■ Strength of Advantage

■ Disadvantage

Fixed Loop Ultra Hi-ThroughputVariable Loop all others (> 98%)

Delay volume overview- Configure your sytem to your needs

Configuration Delay Vol.* / µl

1290 Pump 10

1290 Pump + Fixed Loop1 (for MS) 20

1290 Pump + Jet Weaver + Fixed1 Loop 55

1290 Pump + Variable Loop (for MS) 80

Waters Acquity1 (mixer) 90

1290 Pump + Jet Weaver1 + Variable Loop (for UV) 110

RRLC Pump + Variable Loop (for MS) 260

RRLC Pump + Damper/Mixer + Variable Loop (for UV) 740-940

* Delay Volume: 10 % step gradient Definition, preliminary data1 5 µl Loop

1290 Infinity Thermostatted Column Compartment1290 Infinity Thermostatted Column Compartment

1290 Infinity Therm. Column Compartment- Next level in usability:

New prolonged HX-carrierwith 3rd hand (not shown)

New valve drive and valve head

New capillary guide

New leak funnel

modified lowdisp HX

New door-open sensor

RFID-tag, stores type, pressure range, valve switches

NewThermal insulation

New 2PS/6PT, 2PT/10PT and 9PT/8PS Quick-Change high pressure valves

1290 Infinity LC Solution- Method development and multi-method walk-up solution

Up to 3 clustered TCC

• Up to 8 Columns• Up to 26 Solvents• Up to 6 Temperature Zones

Method development

• Method Scouting Wizard• AutoChrom/ChromSword

Multi-method labs

• ChemStation• Easy Access walk-up

35

1. Highest power range (1200 bar x 2 ml) for highest separation power per time- Speed for higher throughput- Resolution for higher data quality (ion suppression)

2. Lowest delay volume* for fastest gradients(pump delay volume (10 µl w/o mixer*) a) with Fixed loop (20 µl*) injectionb) with Flow through (80 µl*) injection

3. More robustness for lowest carry over with 1290 Infinity Autosampler< 0.002 % Chlorhexidine** * Delay Volume: 10 % step gradient Definition, preliminary Data

1290 Inifinity LC and MS- Infinitely better for LC/MS

June 25, 2009

5x10

0.025

0.05

0.075

0.1

0.125

0.15

0.175

0.2

0.225

0.25

0.275

0.3

0.325

0.35

0.375

0.4

0.425

0.45

0.475

0.5

0.525

0.55

0.575

0.6

0.625

0.65

0.675

0.7

0.725

0.75

0.775

0.8

0.825

0.85

0.875

0.9

0.925

0.95

0.975

1

1.025

1.05

1.075

1.1 63.0Cpd 109: Terbutryn

Counts vs. Acquisition Time (sec)2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 46 48 50 52 54 56 58 60 62 64 66 68 70 72 74 76 78 80 82 84 86 88 90

1290 Infinity LC and MS

(2ppm mass tolerance, MFE)50 pg on-column2.1x50mm C-18

1.8 micronFlow rate 1.5 mL/min

900 Bar

Gradient was t0=10% ACNt1.5=100% ACN

131/220 (60%) Pesticides Identified90 second analysis time

20Hz (2GHz mode)

University of ColoradoDrs. Imma Ferrer and Michael Thurman

5x10

0.040.060.08

0.10.120.140.160.18

0.20.220.240.260.28

0.30.320.340.360.38

0.40.420.440.460.48

0.50.520.540.560.58

0.60.620.640.660.68

0.70.720.740.760.78

0.80.820.840.860.88

0.90.920.940.960.98

11.021.041.061.08 63.0

Cpd 109: Terbutryn

Counts vs. Acquisition Time (sec)2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 46 48 50 52 54 56 58 60 62 64 66 68 70 72 74 76 78 80 82 84 86 88 90

Terbutryn 90 second Analysis time 20Hz

(2GHz mode)

0.7 second Peak width

@ half height 20 Data points across peak

1290 Infinity LC and MS

Agilent 1290 Infinity LC - Infinitely Better for UHPLC and HPLC

MoreFlexible

Best-in-classSensitivity UV : 2 – 20x with 1 cm cell

8 – 80x with 6 cm UHS cellCondition independent

Sensitivity MS/MS: < 1 fg with Jet StreamCarry Over: < 0.002% Chlorhexidine

Supports all analytical columns w/o limitationEnables Method transfer from/to any (U)HPLCFlexible Injection: flow-through and fixed loopHighest-Throughput Solution (> 2000 samples/day)Method Development and Multi-method Solutions: up to 8 columns, 26 solvents, 6 temperature zones

More Sensitive

More Powerful

Best-in-classResolution and Speed: 1200 bar (2 ml/min)

800 bar (5 ml/min)20ul delay volume

RRHD Columns (1.8u / 1200bar): N=50000 in 3min

3rd Party Instrument Control- the concept

• 1120, 1200, 1290 Drivers for all mayor 3rd Party CDS and MS-WS• With identical functionality as Agilent CDS & MS-WS Drivers • Easy plug-in of new modules (“… like a Windows Printer Driver …”)• Developed and sold by Agilent

• Phase 1: Target CY‘09 – Empower, Chromeleon, Analyst• Phase 2: Target CY‘10 – XCalibur, and other CDS and MS-WS• Good progress made. Realization depends on 3rd parties.

Third

Par

ty

App

licat

ion

New

Ada

pter

New

Clip

per S

DK

RC.NET LC Driver

RC.NET LC Driver

RC.NET LC Driver

3rd Party Agilent

Summary: Agilent 1290 Infinity LC - Infinite possibilities

June 06, 2005


Thank you for your attention.

Questions?

agilent 1290 infinity lc · agilent 1290 infinity lc - separation power per time page 12 june 06,...

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