Agilent 1290 Infinity LC · 2016. 8. 30. · Short History of UHPLC 2003 Agilent 1200 Binary 1.8um...
Transcript of Agilent 1290 Infinity LC · 2016. 8. 30. · Short History of UHPLC 2003 Agilent 1200 Binary 1.8um...
June 06, 2005Page 1
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
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What Debate ?
Why is it Over ?
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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.
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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
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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 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
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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 !
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Agilent 1290 Infinity LC- Separation Power per time
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
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Agilent 1290 Infinity LC- Separation Power per time
June 06, 2005Page 12
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
F = 2 ml/minP = 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°CV_i = 0.5 µl WL = 210 nm DR = 80 Hz
10 Compound Pharma Test Mix
1.0min
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Baseline separation of Glafenine diastereomere achieved!Less than 1 minute analysis time!
F = 2 ml/minP = 1085 bar
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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
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Gain N and Rs with RRHD Long Lengths150 +100 mm = 250 mm 1.8um @ 1000 bar
Group/Presentation TitleAgilent Restricted
Month ##, 200XPage 15
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
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1290 Infinity Binary Pump1290 Infinity Binary Pump
June 25, 2009
Agilent Confidential
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1290 Infinity Binary PumpNext generation Pump Technology - How it looks like inside
June 25, 2009Page 18
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)
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1290 Infinity LC – Delay Volume and Mixing Noise- How to achieve lowest delay volume and lowest noise?
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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.
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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
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1290 Infinity Diode-Array Detector1290 Infinity Diode-Array Detector
1290 Infinity Diode-Array Detector- The Next Generation in UV-Vis Detection
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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
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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
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4 x Higher Sensitivity
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1290 Infinity Autosampler1290 Infinity FlexCube
1290 Infinity Autosampler1290 Infinity FlexCube
1290 Infinity Autosampler- Next Generation Variable Loop Injector
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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 ...........
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1290 Infinity FlexCube- Automated needle seat backflushing - Fixed loop injection mode
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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
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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
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1290 Infinity Thermostatted Column Compartment1290 Infinity Thermostatted Column Compartment
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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
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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, 2009Page 36
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
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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
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Agilent 1290 Infinity LC - Infinitely Better for UHPLC and HPLC
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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
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June 06, 2005Page 42
Agilent 1290 Infinity LC
Thank you for your attention.
Questions?