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  • June 06, 2005Page 1

    Agilent 1290 Infinity LC

    Dr. Michael Kraft Pharma Industry Marketing Manager Waldbronn, 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

    Page 2

  • What Debate ?

    Why is it Over ?

    Page 3

  • Short History of UHPLC

    2003 Agilent 1200 Binary 1.8um RRHT Columns 2004 Waters Acquity 1.7um BEH Columns 2006 Agilent 1200 RRLC

    Thermo Accela 2007 Shimadzu UFLCxr

    Jasco Xtreme-LC Hitachi LaChrom Ultra Scient. Syst. Ultra HP

    2008 Dionex RSLC Knauer Platin Blue

    2009 PE Flexar

    10 New UHPLC Systems in 5 Years – All different.

    Page 4

  • 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

    Page 6

    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 Porous TP = 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 Porous TP = Totally PorousPage 7

  • June 06, 2005Page 8

    Agilent 1290 Infinity LC

  • 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

    Page 9

    Advantages Supports all analytical columns

    Enables Method Transfer

    More Resolution

    More Speed

    More viscous solvents

    Lower temperatures

  • Agilent 1290 Infinity LC - Separation Power per time

    . Tranquilizer Sample Column: Superficially porous

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

    Gradient 2-92% ACN in 0.4min T= 80 °C

    0.095sec PW

    0.140secPW

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

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

    Page 10

  • Agilent 1290 Infinity LC - Separation Power per time

    NH

    N H

    O

    O

    OH

    O

    OH NH

    N Cl

    O

    O

    OH

    OH

    Glafenine Peak 3

    O N

    ClH

    Diphenhydramine.HCl Peak 4

    N

    ClH

    Amitriptyline.HCl Peak 5

    O

    O

    O

    O

    O

    O O

    O

    (+)-Bibenzyl-L-tatrate Peak 7

    Diazepam Peak 8

    N

    N Cl

    O

    N H

    OH

    O

    Cl

    Cl

    Diclofenac Peak 9

    Naphthalene Peak 10

    Uracil Peak 1

    Tropic acid Peak 2

    Methyl benzoate Peak 6

    10 Compound Pharma Test Mix

    Page 11

  • Agilent 1290 Infinity LC - Separation Power per time

    June 06, 2005Page 12

    F = 2 ml/min P = 700 bar

    0.6min

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

    F = 2 ml/min P = 700 bar

    10 Compound Pharma Test Mix

    C18 2.1 x 50mm, 1.7 µm

  • Agilent 1290 Infinity LC - Separation Power per time

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

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

    10 Compound Pharma Test Mix

    1.0min

    Page 13

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

    F = 2 ml/min P = 1085 bar

  • 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 µm P_max = 880bar

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

    Page 14

  • Gain N and Rs with RRHD Long Lengths 150 +100 mm = 250 mm 1.8um @ 1000 bar

    Group/Presentation Title Agilent Restricted

    Month ##, 200XPage 15

    α3,2: 1.15 Rs3,2: 6.75 N6 : 49200 Pw2: 0.017 min Pw6: 0.029 min

    α3,2: 1.15 Rs3,2: 4.92 N6 : 29600 Pw2: 0.015 min Pw6: 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. Uracil 2. Acetophenone 3. Propiophenone 4. Butyrophenone 5. Valerophenone 6. Hexanophenone

    1

    2

    6

    5 43

    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

    Page 16

  • 1290 Infinity Binary Pump1290 Infinity Binary Pump

    June 25, 2009

    Agilent Confidential

    Page 17

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

    June 25, 2009Page 18

    Channel A

    Channel B

    Jet Weaver

    Multi-layer Heat exchanger

    Solvent selection valveDegasser

    Purge Valve

    Silicon Carbide Pistons

    High Resolution Pump drives Next Generation Technology

    Highest Speed and Resolution: 1200bar @ 2mL/min 800bar @ 5mL/min Lowest Delay volume: 10 µl / 45 µl Lowest Mixing Noise Highest RT Precision Automatic Purge Automatic 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)

    Page 19

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

    Page 20

    1200 Series Solvent A

    Passive Damper 0-300 ul

    Passive Mixer 400 ul

    1200 Series Solvent B

    1290 Inifinity Solvent A

    1290 Inifinity Solvent B

    Very Good Noise ~ 800 µl

    + = Active Damping

    10µl Microfluidic 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.

    Page 21

  • 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, 1.8µm Sample: Phenone Standard

    Injection volume: 0.5ul Mobile phase: H2O/ACN with Jetweaver

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

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