new advances in hplc - agilent · (with flexible cube for lowest carry-over) 1290 infinity...
TRANSCRIPT
![Page 1: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/1.jpg)
New Advances in UHPLC-Resolution, Speed & Sensitivity
Enhancing Productivity with Agilent’s Newest Instrumentation
Page 1
Patrick CronanLC Applications Scientist
Boston, MA
September 14, 2012
![Page 2: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/2.jpg)
The 1290 Infinity Quaternary LC SystemHow binary pump-like performance can enhance your lab efficiency
1290 Infinity DAD with 60mm cell (& HDR optional)
1290 Infinity TCC with broadest
Page 2
September 24, 2012Page 2
1290 Infinity TCC with broadest valve selectivity
1290 Infinity Autosampler (with Flexible Cube for lowest carry-over)
1290 Infinity Quaternary Pump
![Page 3: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/3.jpg)
Innovation: 1290 Infinity Quaternary PumpQuaternary Pump with Binary Pump-like Performance
1290 Infinity Quaternary Pump
• Boost PerformanceHighest accuracy and precision for composition and flow, with exceptionally low delay volume
Page 3
• Save TimeAccelerates transfer of existing HPLC methods to UHPLC
• Reduce CostsOutstanding UHPLC performance at HPLC-like costs of ownership
![Page 4: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/4.jpg)
New Inlet-Mixer - based on JetWeaver technology
Page 4
Oct. 2011
4
![Page 5: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/5.jpg)
1290 Infinity Quaternary PumpVarious tools enabling the high performance
Multi-Purpose-
Switchable Jet Weaver Mixer
Page 5
5
4-channel Degasser
MCGV (Multi-Channel-Gradient-Valve) Inlet-Weaver Pump Head
Multi-Purpose-Valve
In-line Filter
![Page 6: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/6.jpg)
Multipurpose valve functions for highest comfortStandard Application for standard Delay Volume
Page 6
6
![Page 7: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/7.jpg)
Multipurpose valve functions for highest comfortExtra Mixing Volume for lowest baseline ripple (TFA applications)
Page 7
7
![Page 8: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/8.jpg)
Multipurpose valve functions for highest comfortAutomatic Purging
Page 8
8
![Page 9: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/9.jpg)
Multipurpose valve functions for highest comfortAutomatic filter backflush
Page 9
9
![Page 10: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/10.jpg)
Power Range� For any kind of analysis
Composition Accuracy and Precision< 0.15 % RSD or 0.02 min SD
� 0.4 % (1-99 % Composition B)
� High RT precision in gradient runs
Agilent 1290 Infinity Quaternary PumpSpecifications & Benefits
Page 10
gradient runsFlow Accuracy and Precision
< 0.07 % RSD or 0.01 min SD
� 1.0 % or 10 µL
� High RT precision in isocratic runs
Composition Range1-99 %
� Wide analytical range
Delay Volume< 350 µL
� For fast quaternary gradients
![Page 11: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/11.jpg)
� Different mixtures (H2O, MeOH, ACN) at � different pressures and � different flow rates
Full application range!
Composition Accuracy and PrecisionStep Gradient Analysis with Tracer
Page 11
11 January, 26, 2012
![Page 12: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/12.jpg)
Flow and Composition Accuracy and PrecisionIsocratic analysis of Glucocorticoids
3: Hydrocortisoneacetate
1: Hydrocortisone 2: Beclomethasone-di-proprionate
Page 12
Column: Agilent ZORBAX Eclipse Plus C18, 4.6 x 100 mm, 5 µm (p/n 959996-902)
Mobile phase: Water/ Acetonitrile = 30/70Flow rate: 1 mL/min
Stop time: 5 min
Injection vol.: 3 µL (needle wash for 3 sec)
Column Temp.: 30 °C
DAD: 254/10 nm, (ref 400/ 80 nm), 20 Hz, slit 4 nm
![Page 13: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/13.jpg)
Flow and Composition Accuracy and PrecisionIsocratic analysis of Glucocorticoids
mAU
25
30
1
2
RT of compounds very sensitive regarding changes in
mobile phase composition!
Page 13
min1 1.5 2 2.5 3 3.5
0
5
10
15
20
Water/ACN 29:71
3
Water/ACN 31:69
Water/ACN 30:70
![Page 14: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/14.jpg)
Accuracy and PrecisionIsocratic analysis of Glucocorticoids
mAU
35
1: Hydrocortisone
Specification (< 0.07 %)
Page 14
min0 1 2 3 4
0
5
10
15
20
25
30
35
3: Hydrocortisoneacetate
2: Beclomethasone-di-proprionate
![Page 15: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/15.jpg)
Power Range� For any kind of analysis
Composition Accuracy and Precision< 0.15 % RSD or 0.02 min SD
� 0.4 % (1-99 % Composition B)
� High RT precision in gradient runs
Agilent 1290 Infinity Quaternary PumpSpecifications & Benefits
Page 15
gradient runsFlow Accuracy and Precision
< 0.07 % RSD or 0.01 min SD
� 1.0 % or 10 µL
� High RT precision in isocratic runs
Composition Range1-99 %
� Wide analytical range
![Page 16: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/16.jpg)
Composition RangeShallow gradient at low organic concentration
1: Theophylline
N
N
N
N
CH3
O
CH3
O
H
N
N
N
N
CH3
O
CH3
O
CH3
2: Caffeine
Page 16
Column: Agilent Poroshell 120 EC-C18, 4.6 x 50 mm, 2.7 µm (p/n 699975-902)
Mobile phase: A = Water / B = Acetonitrile
Gradient: 2 – 12 % B in 5minFlow rate: 2 mL/min
Stop time: 5 min
Injection vol.: 5 µL
Column Temp.: 40 °C
DAD: 254/10 nm, (ref. 360/ 100 nm), 20 Hz
![Page 17: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/17.jpg)
Composition RangeShallow gradient at low organic concentration
mAU
80
100
1: Theophylline
N
N
N
N
CH3
O
CH3
O
H
N
N
N
N
CH3
O
CH3
O
CH3
2: Caffeine
1
Page 17
min2 2.5 3 3.5 4 4.5
0
20
40
60
2
![Page 18: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/18.jpg)
Composition RangeShallow gradient at low organic concentration
0.3
0.4
0.5
RS
D R
T (%
)Retention time precision
Page 18
0
0.1
0.2
0.3
1290 InfinityQuat. System
RS
D R
T (%
)
Theophylline
Caffeine
Specification (< 0.15 %)
![Page 19: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/19.jpg)
Composition RangeShallow gradient at low organic concentration
0.3
0.4
0.5
RS
D R
T (%
)Retention time precision
2.4
Page 19
0
0.1
0.2
0.3
1290 InfinityQuat. System
1260 InfinityQuat. System
Quat. UHPLCOther vendor
RS
D R
T (%
)
Theophylline
Caffeine
Specification (< 0.15 %)
![Page 20: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/20.jpg)
Composition RangeShallow gradient at low organic concentration
mAU
200
250
Step gradient1 % steps from 0 to 10 %
Page 20
0 10 20 30 40 50
0
50
100
150
1290 Infinity Quaternary system
Quaternary UHPLC from other vendor
![Page 21: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/21.jpg)
Ternary/Quaternary GradientsBlend Assist
You need different concentrations of modifiers in your analysis, would like to have just one stock-solution and do online dilution to profite from the quaternary mixing capability of your pump? Here is a simple tool – BlendAssist!
Water Water 1% TFA
Desired method conditions - example: 1. 5 to 95% gradient of ACN with 0.1% TFA in
Water and 0.08% TFA in ACN2. 20 – 80% gradient of ACN with 0.5% TFA in
Page 21
1% TFA
ACNACN
1% TFA
Water and 0.4% TFA in ACN
Without BlendAssist you need to either pre-mix the required solvents or by using stock-solutions of TFA in Water and ACN to program complex gradients (%A, B, C, D).
With BlendAssist: just program your binary organic/aqueous gradient and define the dilution factor!
![Page 22: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/22.jpg)
Ternary/Quaternary GradientsBlend Assist - simple Tool for Online-Dilution of Modifiers and Gradient Set-Up
Page 22
BlendAssist - Solvent/modifier channel definition
![Page 23: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/23.jpg)
Buffer Advisor SoftwareQuaternary mixing
Page 23
September 24, 201223
![Page 24: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/24.jpg)
1260 Infinity Bio-inert HPLC
What is similar compared to standard1260 Infinity Quaternary LC?
� Pumping performance, gradient performance
� Pumping Specifications� Detector, Autosampler Performance and
Page 24
� Detector, Autosampler Performance and Specs
� Overall System Robustness� TCC specs
600 bar also for fast or high resolution Bioseparations
Modular flexibility maintained
September 24, 201224
![Page 25: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/25.jpg)
1260 Infinity Bio-inert HPLC
What is different compared to standard 1260 Infinity Quaternary LC
� Pump titanium based, no SST �Autosampler metal-free�Capillaries and connections metal-free�Detector Flow Cell metal-free
Page 25
�Detector Flow Cell metal-free�pH-range 1-13, short term pH 14� Active Seal wash included in pump� Low pressure is possible
•Wide pH range•High salt tolerance•Lowest surface activity� Bio-Inertness
September 24, 201225
![Page 26: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/26.jpg)
New Capillary Design Ensures Bio-inertnessCapillaries:
Metal cladded PEEK capillary
New capillary technolgy enables 600 bar
AND is completely metal free !!
Page 26
September 24, 201226
Needle:
SST housingfor robustness
PEEK and ceramic for metal free sample path
![Page 27: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/27.jpg)
Page 27
![Page 28: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/28.jpg)
Page 28
![Page 29: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/29.jpg)
Buffer Advisor SoftwareWorkflow
1: Select Buffer and Gradient Solution
2: Enter Gradient Timetable
3: Review Composition
Page 29
September 24, 201229
3: Review Composition of Stock Solutions and Adjust
4: Gradient Display Section: Process and Review Gradient
![Page 30: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/30.jpg)
Experimental Situation
Page 30
![Page 31: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/31.jpg)
Buffer Advisor SoftwareWorkflow
Page 31
![Page 32: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/32.jpg)
Buffer Advisor SoftwareTimetable import into Chemstation
Page 32
32
![Page 33: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/33.jpg)
Buffer Advisor SoftwarepH scouting
Page 33
33
![Page 34: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/34.jpg)
Buffer Advisor Softwarestable pH
Page 34
34
![Page 35: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/35.jpg)
30x wider linear UV range -Quantification of widely different concentration levels in one single run
Page 35
Agilent 1200 Infinity High Dynamic Range (HDR) Solution
![Page 36: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/36.jpg)
History of DAD Linearity Gain The last 30 years
> 30
20.0
25.0
30.0
35.0
40.0
45.0
Line
arity
Gai
n
1200 Infinity HDR-DAD
With HDR
>30X Linearity
Page 36
Detector Model (Intro Year)
1.4 1.7 2.0 1.4 2.9
0.0
5.0
10.0
15.0
20.0
1040 to 1090 1090 to 1050 1050 to 1100 1110 to 1200 1200 to 1290/60 1200 to HDR
Line
arity
Gai
n
(1988)(1986) (1995) (2006) (2010) (2011)
September 24, 201236
![Page 37: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/37.jpg)
Programmable
GratingMirror
Optofluidic Waveguides: Max-Light Flow Cells Total-internal reflection in a non-coated fused silica fiber
1260 / 1290 Infinity DAD
Page 37
Deuterium Lamp
Max-Light Cartridge Cell10 mm or 60 mm path length
Programmableor fixed slit
1024 elementdiode-array
September 24, 201237
![Page 38: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/38.jpg)
Effects of path length increase
Peak Dispersion
60 mm pathlengthlarge geom. cell volume (78 µL)
- Loss of resolution- Loss of signal height
10 mm pathlengthsmall geom. cell volume (13 µL)
high light transmission
Conventional flow cells:
Page 38
large geom. cell volume (78 µL)lower light transmission
60 mm pathlength4 µL �V dispersion volume
Max-Light High Sensitivity cell:
Optofluidic waveguides (total internal reflection)
September 24, 201238
![Page 39: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/39.jpg)
10x Higher Sensitivity 1290/1260 Infinity DAD compare to 1200 Series DAD SL
mAU
20
25
30
Columns: 150 x 4.6mm Zorbax SB C18, 5µm
Sample: Anthracene: 835 pg/µL
Mobile phase: A: Water, B: Acetonitrile
Elution: isocratic 80 % B
Injection volume: 5 µL
Flow: 1.5 mL/min
DAD: 251/4nm, Ref= 450/80nm, 2.5Hz, slit width 4nm
1260/1290 Infinity DAD
Page 39
1260/1290 DAD60 mm
1200 DAD SL 10mm
Height [mAU] 28.876 4.938
Noise [mAU] 0.0098 0.0190
Signal/ noise 2944 259
Increase 11.4 xmin0 1 2 3 4 5
0
5
10
15
20 DAD: 251/4nm, Ref= 450/80nm, 2.5Hz, slit width 4nm
1200 Series DAD / DAD SL
September 24, 201239
![Page 40: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/40.jpg)
30x Wider Linear Range with HDR-DAD3.7 mm and 60 mm and Max-Light flow cell
3.7 mm path-length
Detector Signal[AU/cm]
Page 40
60 mm path-length
Computing the signals from60 mm path-length for the low concentration3.7 mm path-length for the high concentration
Conc [µg/mL]
September 24, 201240
![Page 41: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/41.jpg)
1290/1260 High Dynamic Range – HDR Increasing Linear UV Range by >30x
Technology
� Cluster of two DADs
� DAD 1: 60 mm Cell – for low concentrations
� DAD 2: 3.7 mm Cell – for high concentrations
� Output: One, combined signal, normalized to 10 mm path length (HDR range: 0.6 x 10-6 to 6.7 AU/cm) vs for 1200 Series: 7 x 10-6 to 2 AU/cm)
60mmDAD 1:
Page 41
Control / Usability
� Like standard 1290/1260 DAD: Control, data analysis, reporting
Investment Protection
� Existing 1290/1260 DAD can be upgraded (by 2nd DAD)
� Existing 1100/1200 Systems can be upgraded (by two 1290 or 1260 DAD)3.7mm
Clustered DADDAD 2:
September 24, 201241
![Page 42: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/42.jpg)
Fixed Dose CombinationsFrom Roche Switzerland
Fixed dose combinations:
A combination of two or more
APIs in a fixed ratio of dosesAgilent 1200 (black): API 2: 10 µl injected
Agilent 1200 (red): API 1: 100 µl injected
Infinity HDR-DAD (blue): API 1 and 2 simultaneously: 70 µl injected
Page 42
API 1
API 2
September 24, 201242
![Page 43: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/43.jpg)
Fixed Dose CombinationsFrom Roche Switzerland
“With the Agilent 1200 Infinity HDR-DAD we significantly decreased our turnaround time. We also get lower
LODs and better area precision”
Dr. Rose-Marie Meier F. Hoffmann - La Roche AG
Page 43
Infinity HDR-DAD
Agilent 1200 DAD
September 24, 201243
![Page 44: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/44.jpg)
2 Injections needed if a conventional DAD is used
1µl injection to determine the first 3 peaks
min0 1 2 3 4 5
mAU
0
200
400
600
800
1000 Vitamin C
Paracetamol
Caffeine Chlorphenamine ?
Page 44
September 24, 201244
Vitamin CParacetamol
Caffeine
Chlorphenamine
5µl injection to determine the last peak and further impuritiesThe first 2 peaks are out of linear range
min0 1 2 3 4 5
mAU
0
1000
2000
3000
3800Detector saturationOutside the linear range
![Page 45: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/45.jpg)
2 Injections needed if a conventional DAD is used
ChlorphenaminemAU
15
17.5
20
DAD1 A, Sig=254,20 Ref=380,80 (SOP 98 GRI...0FEB\HDR_GRIPPO_OLD_20FEB 2012-02-20 10-40-57\GRIPPO_1UL09.D)
DAD1 A, Sig=254,20 Ref=380,80 (SOP 98 GRI...0FEB\HDR_GRIPPO_OLD_20FEB 2012-02-20 10-40-57\GRIPPO_5UL10.D)
mAU
0
0.2
0.4
0.6
0.8
1
Chlorphenamine
Imp 3
Imp 4
Page 45
September 24, 2012Confidentiality Label
45
Blue: 1µl injection with 3.7mAU peak height for Chlorphenamine
Red: 5µl injection with 21.8mAU peak height for Chlorphenamine
min4 4.5 5 5.5 6
0
2.5
5
7.5
10
12.5
15
Imp 3 Imp 4
min4.6 4.7 4.8 4.9 5 5.1 5.2 5.3
-0.2
0
![Page 46: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/46.jpg)
Comparison of conventional DAD vs. HDR DAD, 5µl injection
mAU
2500
3000
3500
4000
Red conventional DAD: 2 peaks out of linear range with 5µl injection
Green HDR-DAD: All peaks within linear range
Paracetamol
Vitamin CDetector saturationOutside the linear range
Page 46
September 24, 201246
min0 1 2 3 4 5
0
500
1000
1500
2000All peaks within linear range (up to 6AU)
Caffeine
Chlorphenamine
![Page 47: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/47.jpg)
Comparison of LOD
mAU
0.8
0.9
1
1.1
1.2
1.3
HDR DAD
Conventional DAD
More reliable automated peak integration
Page 47
September 24, 201247
Compound LOD with S/N=3 for Conventional DAD
LOD with S/N=3 for HDR- DAD solution
Chlorphenamine ~1ng ~0.1ng
min3.9 4 4.1 4.2 4.3 4.4 4.5 4.6 4.7 4.80.5
0.6
0.7
0.8
![Page 48: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/48.jpg)
Comparison of conventional DAD and HDR DAD, 5µl injection
mAU
15
20
DAD1 K, Sig=254,20 Ref=380,80 (SOP 98 GRI...B\HDR_GRIPPO_HDR_21FEB 2012-02-21 07-51-02\GRIPPO_HDR5UL09.D)DAD1 A, Sig=254,20 Ref=380,80 (SOP 98 GRI...0FEB\HDR_GRIPPO_OLD_20FEB 2012-02-20 10-40-57\GRIPPO_5UL09.D)
Red conventional DAD: determination of impurities and chlorphenamine in a second run
Blue HDR DAD: dermination of all peaks in one run
ParacetamolCaffeine
ChlorphenamineVitamin C
Page 48
48
min1 2 3 4 5
0
5
10
Imp 1 Imp 2 Imp 3 Imp 4
![Page 49: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/49.jpg)
Comparison of area precision
14.908 13.013 5.37 5.723
1
1.5
2
2.5
3
RS
D A
rea
(%)
Comparison of Area Precision
HDR Signal
Conv.DAD signal (1µl)
Conv.DAD (5µl)
Page 49
September 24, 201249
HDR DAD 5µl injection volume for detemination of all compounds (blue) Conventional DAD: 1µl injection volume for determination of Vit C, paracetamol and Caffeine (orange)
5µl injection volume for determination of Chlorphenamine and further impurities (green)
0
0.5
![Page 50: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/50.jpg)
Intelligent System Emulation Technology
(ISET)
Page 50
September 24, 2012Page 50
![Page 51: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/51.jpg)
What is ISET?- Two components
A. Best in class performance of the 1290 Infinity LCBroad power range, ultra-low delay volume, superior sensitivity, unmatched flow and composition accuracy
Peak 3Overlay of 10 runs
RSD = 0.005 %
Page 51
Page 51
B. The revolutionary ISET emulationalgorithmFull characterization of target LC systemsCreation of emulation function
FA
![Page 52: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/52.jpg)
Method Transferability: 1290 Infinity LC- System Emulation Technology
Concept
EmulatedSystem A
FA
Page 52
Page 52
Select System to be emulated by a simple mouse click:
Agilent 1100 Series
Agilent 1200 Series LC
Agilent 1260Infinity LC
Agilent 1220Infinity LC
Alliance(2012)
![Page 53: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/53.jpg)
Intelligent System emulation technology- How it works (1290 to 1200)
250
300
350
400
mAU
Injection
Same gradient conditions by moving the gradient and by emulating the gradient mixing behavior.
Page 53
Page 53
Programmed gradient
1290 gradient
1200 gradient
0 0.5 1 1.5 2 2.5 3 3.5
0
50
100
150
200
min
gradient mixing behavior.
![Page 54: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/54.jpg)
Select pump
Intelligent System Emulation Technology ISETImplementation
Page 54
Select sampler
Page 54
![Page 55: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/55.jpg)
PesticidesColumn: 3 x 50 mm, 2.7 µm Poroshell Run time: 3.5 min
mAU
500
600
700
800
Sim
azin
e
Chl
orid
azon
e
Chl
orto
luro
n
Diu
ron
Pro
pazi
neTe
rbut
ylaz
ine
Pro
met
ryn
1290 System with iSETM
etam
itron
Cya
nizi
ne
Rs= 1.87
Page 55
55
min1 1.5 2 2.5 3 3.5
0
100
200
300
400
1290 System
Rs= 1.90
1100 Binary System
Rs= 1.15
![Page 56: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/56.jpg)
PesticidesColumn: 3 x 50 mm, 2.7 µm Poroshell Run time: 3.5 min
Page 56
56
![Page 57: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/57.jpg)
PesticidesColumn: 3 x 50 mm, 2.7 µm Poroshell Run time: 3.5 min
Page 57
57
![Page 58: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/58.jpg)
Generic emulation- Limitations
Example: 1290 to Vendor X
Injection
250
300
350
Generic emulation: Move the gradient
mAU
Page 58
Page 58
0 0.5 1 1.5 2 2.5 3 3.5
0
50
100
150
200
min
Programmed gradient
1290 gradient
Vendor X
![Page 59: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/59.jpg)
Generic emulation: Move the gradient
Generic emulation- Limitations
Example: 1290 to Vendor X
Injection
250
300
350
mAU
Generic emulation: Move the gradient
Page 59
Page 59
Programmed gradient
1290 gradient
Vendor x
0 0.5 1 1.5 2 2.5 3 3.5
0
50
100
150
200
min
Does not consider the gradient mixing behavior (different curves) ordelay volume changes because of pressure changes for pumps with damper.
![Page 60: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/60.jpg)
Method Transfer Possibilities Today- A) Adding physical volume (plumbing solution)
1290 + 1000 �L dwell vol.
Example: 1290 Infinity LC
1100/1200 Quat Pump
Page 60
Page 60
Advantages
• Works well for generic, slower analysis• No Method change
Disadvantages
• Requires hardware change, no flexibility
• Hardware change difficult in validated environment
• Requires manual determination of the dwell volume (Pumps with damper the dwell volume is not constant and depends on pressure)
![Page 61: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/61.jpg)
Isocratic hold: move the gradient
Method Transfer Possibilities Today- B) Isocratic Hold
Example: 1290 to 1200
Injection
250
300
350
mAU
Isocratic hold: move the gradient
Page 61
Page 61
Programmed gradient
1290 gradient
1200 gradient
0 0.5 1 1.5 2 2.5 3 3.5
0
50
100
150
200
min
Does not consider the gradient mixing behavior (different curves).
![Page 62: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/62.jpg)
Method Transfer Possibilities Today- B) Isocratic Hold
Example: 1290 to 1200
1200 Series LCBinary Pump
1290 Infinity LC+ 860 µl “Delay Volume”
Page 62
Page 62
1290 Infinity LC+ 900 µl “Delay Volume”
1290 Infinity LC+ 940 µl “Delay Volume”
![Page 63: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/63.jpg)
Method Transfer Possibilities Today- B) Isocratic Hold
Example: 1290 to 1200
1260 Infinity LC
1290 Infinity LC+ 900 µl hold
Page 63
Page 63
Advantages
• Flexibile, Isocratic Hold is part of the method
Disadvantages
• Works not in all cases• Requires manual determination of the dwell volume/
isocratic hold (Pumps with damper the dwell volume is not constant and depends on pressure)
• Requires modification of the methods (should be avoided in validated environment,but doesn’t require revalidation USP Chapter <621>)
![Page 64: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/64.jpg)
Method transfer, pesticide example2.1x100 mm Zorbax Eclipse Plus, 1.8 µm column, flow: 0,8 mL/min
(A)1100
Quaternary400 bar
(B)
1 2,3 4 5 6,7 8 9 10,11,12 13 14 15 16
Page 64
September 24, 2012Page 64
(C)1290 Infinity
1200 barwith ISET
(B)1290 Infinity
1200 bar
![Page 65: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/65.jpg)
Summary1290 Quaternary Pump:� Full 1290 Infinity LC Power range
� Highest composition precision and accuracy across pressure and flow rate range
� Full application range with binary-like pump performance! 1290 Systems using ISET:• Achieves higher productivity for method development • Simplify your instrument to instrument method transfer while minimizing costs
• Run your legacy methods with ISET and use UHPLC speed for faster separations
Page 65
• Run your legacy methods with ISET and use UHPLC speed for faster separations
HDR DAD Solution:• Determination of low and high dose drugs is possible in one run• Improved Precision and Lower Detection Limits as compared to convention DAD
Using Buffer Advisor with BioInert Quat System:• Shortens and simplifies the workflow for pH scouting by using pre-validated
buffer systems and dynamically mixed gradients• Excellent tool using for automated pH scouting and accurate ion exchange
chromatography
65
![Page 66: New Advances in HPLC - Agilent · (with Flexible Cube for lowest carry-over) 1290 Infinity Quaternary Pump. Innovation: 1290 Infinity Quaternary Pump ... Buffer Advisor Software Timetable](https://reader034.vdocuments.mx/reader034/viewer/2022042808/5f871efe7a8f3b43987ec5b1/html5/thumbnails/66.jpg)
Page 66