Page 1
The Right Solution:
The Agilent 1200 Infinity
Series for LC Analysis
Laura Phillips
Application Scientist
Agilent Technologies
June 27th, 2013
Page 2
The Agilent 1200 Infinity Series Where Do We Come From?
June 27, 2013
1100 Bin/Quat
HPLC
Bin (HPM)
Quat (LPM)
400 bar
5 mL/min
1290 Infinity (2009)
UHPLC
Bin (HPM)
Quat (LPM - 2012)
1200 bar
5 mL/min
1260 Infinity Bin/Quat
UHPLC
Bin (HPM) – 5mL/min
Quat (LPM) – 10mL/min
600 bar
Price ~ 1200 Bin/Quat
1220 Infinity
UHPLC
Bin (LPM)
600 bar, 5 mL/min
Price < 1200 Quat
1200 Infinity Series
1200 Bin/Quat
HPLC
Bin (HPM)
Quat (LPM)
400 bar
5 mL/min
1200 RRLC
UHPLC
Bin (HPM)
600 bar
5 mL/min
Price: 1200 Bin+20%
1200 Series (2006 – 2010)
1100 Series (1995 – 2006)
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The Agilent 1200 Infinity Series Raising the Standard in HPLC – UHPLC for EVERY Lab
June 27, 2013
1200 Infinity Series
1220 Infinity
Isocratic, manual
Gradient, manual
Gradient, automated
Gradient-DAD
1260 Infinity
Low-flow
Cap.LC
Nano-LC
Chip-LC
Analytical flow
Binary
Quaternary
Isocratic
Binary-SFC
Quaternary Bio-Inert
Prep-flow
1290 Infinity
Binary
Quaternary
new
new
Page 4
The Agilent 1200 Infinity Series Raising the Standard in HPLC – UHPLC for EVERY Lab
June 27, 2013
Method Developement
Solution
Multi-method Solution
High-Dynamic Range DAD
2D-LC Solution
Online-SPE Valve solutions Bioinert
solutions SFC solutions
GPC solutions
Page 5
1220 Infinity LC A solution to all your needs
• Dual gradient pump with integrated
degasser, 600 bar, up to 10 mL/ min, for
flexible and reliable analysis
• Vial sampler with injector programming for
higher automation
• 80°C oven for one 25 cm column for better
results
• Variable wavelength or diode array
detector with 80 Hz sampling rate for high
sensitivity
June 27, 2013
• Easy: Small footprint, low height. One
cable.
• Affordable with ChemStation/ EZChrom
VL software
• Expandable: Any other module can be
added
Page 6
600 bar for HPLC and UHPLC
June 27, 2013
Same as with the 1260 Infinity LCs, existing conventional HPLC methods
from 1100/1200 Series LC can be run on the 1220 Infinity LC and giving the
same results as well as UHPLC methods on 3.0mm and 4.6mmID columns
can be run. Both with excellent performance!
Saving half of the time by transferring a HPLC
method to UHPLC, without loss of resolution. Excellent retention and area RSDs even under
UHPLC conditions.
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Expandability
June 27, 2013
External tray for walk-up:
Together with Agilent‘s EasyAccess software the 1220 Infinity LC with external
tray is a perfect solution for walk-up environments!
Additional detectors:
Almost all 1260/1290 Infinity detectors and SQ-MS can be added!
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Innovation: 1220 Infinity Mobile LC Solution Affordable, Flexible, Compatible – Everywhere!
June 27, 2013
Instrument Control Framework
• Boost Performance
Complete UHPLC system for
higher resolution and shorter
run times
• Save Time
Bring your laboratory to the
samples
• Reduce Costs
Most affordable system
giving you access to UHPLC
technology
Page 9
1260 Infinity LC
1260/1290 Valve Solutions
1260 Infinity Bio-Inert LC – Buffer Advisor
1260 Infinity SFC
1260 Infinity GPC
1260/1290 ELSD
June 27, 2013
Page 10
1200 Infinity Series valve solutions
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NEW Quick-Change Valve heads
for universal Valve Drive
1290 Infinity TCC 1290 Infinity Flexible Cube 1290 Infinity Valve Drive
Types 600 bar, 1200bar
Std, Bio-inert, µ-flow, prep
2pos/4port duo
2pos/ 6port
2pos/10port
4pos/10port
6pos/14port
8pos/ 9port
12pos/13port
6pos/ 7port
2pos/ 8port
2pos/12port
Custom Valves
Applications • Multi-method/Method Dev
• Column switching (2,4,6,8)
• Solvent selection (13 – 26)
• Matrix-stripping
• Sample Enrichment
• Online-SPE
• Detector selection
• Heart Cutting 2DLC
• Comprehensive 2DLC
• Alternating Column Regen.
• Process Control/ Sample
Sampling/ Fractionation
• Column Connection
The largest and most flexible valve portfolio Workflow- and Application-Automation Flexibility
July 30, 2013 11
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Automated Column Regeneration 2-position / 10-port valve head
• 1x 1290 Infinity
Thermostatted column
compartment (or 1290 Infinity
Valve Drive)
• 1x 2-pos/10-port valve head
• While the analysis is running
on column 1, column 2 is
regenerated using the flow
from a second pump
• After the analysis on column
1, the flow switches to
column 2 for analysis while
column 1 is regenerated.
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Sample Enrichment 2-position / 6-port valve head
• 1x 1290 Infinity
Thermostatted column
compartment (or 1290 Infinity
Valve Drive)
• 1x 2-pos/6-port valve head
• The analyte is retained and
concentrated on an
enrichment column while the
sample matrix flows to
waste.
• For analysis, the
concentrated sample is
flushed out of the
enrichment column to the
analytical column.
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Sample Cleanup 2-position / 6-port valve head
• 1x 1290 Infinity
Thermostatted column
compartment (or 1290 Infinity
Valve Drive)
• 1x 2-pos/6-port valve head
• Matrix components are
retained on a pre-column
while the analytes pass
through to the analytical
column for separation.
• After sample cleanup or
matrix stripping, the
precolumn is flushed in
reversed flow direction by
the second pump.
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Column Selection for Method Development 6-position / 14-port valve head
• 1x 1290 Infinity
Thermostatted column
compartment
• 1x 6-pos/14-port valve
head
• Up to 6 columns (4 columns
recommended because of
available space)
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Solvent Selection for Method Development 12-position selection valve head
Typically used with 1290 Infinity Valve Drive.
For Method Development systems:
1290 Infinity Valve Drive and new 12-position
selection valve head is fully supported
as pump-valve cluster and by Agilent
ChemStation Method Scouting Wizard
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1260 Infinity Bio-Inert LC
June 27, 2013
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Innovation: 1260 Infinity Bio-inert LC Truly Bio-inert UHPLC System
June 27, 2013
1260 Infinity Bio-inert LC
• Boost Performance
Highly versatile (U)HPLC
system for small- and large-
molecule analysis
• Save Time
Fast switching between bio-
and non-bio applications,
including cleaning
• Reduce Costs
Only one system to maintain
for bio- and non-bio analysis
Stainless steel system
1260 Infinity Bio-inert LC
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New Capillary Design Ensures Bio-inertness
June 27, 2013
Capillaries
Metal clad PEEK capillary
Newest fitting design: Hybrid technique
New capillary technolgy enables
600 bar AND is completely metal free !!
Page 20
June 27, 2013
Buffer Advisor Software Facilitates Workflows in Ion Exchange Chromatography
Buffer Advisor helps to:
1. Define stock solutions for salt and pH
gradients
2. Check if the buffer system is suitable
for your separation task
3. Create a corrected time table for a
salt- or pH gradient.
Manual preparation:
1. Tedious
2. Error-prone
3. Irreproducible (operator)
Part of Bio-inert pump,
G5611A
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Buffer Advisor User Interface
Area 1: Select Buffer and
Gradient Type
June 27, 2013
Area 2: Enter Gradient
Time Table
Area 4: Review
Composition of Stock
Solutions and Adjust
Area 3: Gradient Display
Section: Process
Gradient and Review
Optimized Gradient with
Compensation Steps
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1260 Infinity SFC
June 27, 2013
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Innovation: Hybrid SFC/UHPLC System Double your Discovery
June 27, 2013
Hybrid SFC/UHPLC System
• Boost Performance
SFC with LC-like sensitivity,
unique 600 bar x 5 mL/min
power range
• Save Time
Efficient instrument utilization
and orthogonality through
unique switching capability
between LC and SFC
• Reduce Costs
Lower solvent expenses, less
waste, and only one system
to maintain. Upgrade from LC
to SFC
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Agilent 1260 Infinity Analytical SFC/MS System 1260 Infinity SFC and the G6100 SQ Mass Spectrometer
June 27, 2013
Analysis of leachables in
food packaging material by
SFC/MS
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1260 Infinity GPC
June 27, 2013
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High Performance GPC/SEC for Routine Work
June 27, 2013
High Performance GPC/SEC Solution
• Boost Performance
High performance pump & oven
for reproducible molecular
weights
• Save Time
RID warm up less than 1 hour,
complete system based on
industry-leading Agilent LC
systems for maximum uptime
• Reduce Costs
RID with built-in solvent recycle,
minimum training effort.
“I have always been impressed by the
quality, robustness, reliability and ease-
of-use of their products, which have
served me well.” - John P. O’Donnell,
Research Scientist, FUJIFILM Imaging Colorants
Ltd., Manchester, UK
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1260 Infinity GPC/SEC Multi Detector Suite Summary
June 27, 2013
Key Benefits
Comprehensive Allows complex multi detector GPC
experiments to be performed routinely.
Versatile The MDS can be controlled by software
or the keypad for maximum ease of
use, and can be used with any existing
modular or integrated HPLC system.
Convenient Data from an existing detector can be
collected and processed through the
MDS.
High Quality
Results
The detector cells are heated for
optimum baseline stability and
minimum instrument downtime.
Adaptable Can be expanded at any time to
include additional detectors.
Safe Incorporates multiple solvent leak
detectors to prevent spillage of
potentially hazardous solvents.
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Agilent 1260 Infinity ELSD
& 1290 Infinity ELSD
Evaporative Light Scattering
Detection (ELSD)
Page 29
Introduction: ELSD General Benefits
June 27, 2013
• Universal detection
• More uniform response
• Gradient Compatible
• Complimentary to LC-MS
• Eliminates derivatization steps in analytical methods
Page 30
Agilent ELSD Portfolio
June 27, 2013
G4260B G4261B
1260 Infinity ELSD
1290 Infinity ELSD
(Cooling deleted)
1290 Infinity ELSD
(Standard, cooled)
Light Source Blue LED (480nm) Blue Laser (405nm)
Temperature
range
Nebulizer: 25-90°C
Evaporator: ambient – 120°C
Nebulizer: 25-90°C
Evaporator: ambient – 120°C
Nebulizer: 25-90°C
Evaporator: 10°C – 80°C
Features
• Hydrocortisone limit of detection
30ng
• RS232 Communication only
• Hydrocortisone limit of detection 4ng
• 5-9 x improvement in sensitivity over current model
• Unique sub-ambient operation (greater response for volatile
compounds)
• RS232 and LAN Communication
• Voltage selection (110/240V) no longer required
• Variable gas flow for uniformity of response
• Fast data collection (up to 80Hz)
• Controlled via ChemStation / EZChrom
• Front Panel control of all parameters (Temperature, gas flow, data rate etc)
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Increased sensitivity, reduced baseline noise
June 27, 2013
The increased laser intensity
coupled with a high gain
photomultiplier and digital signal
processing increases signal and
reduces noise.
G4261B
S/N 158
G4260B
S/N = 5
Injection: 10µL (Hydrocortisone 50ng-on column)
Column: Zorbax Eclipse XDB-C18 (5µm, 4.6x150mm)
Mobile Phase: Water/ACN (80/20)
Flow Rate: 1.0 m/min
ELSD: Neb 40°C, Evap 60°C, Gas 1.05SLM
Signal/noise calculated using OpenLab Chemstation v1.04
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Triglycerides in Natural Oils
June 27, 2013
Sample: 2mg/ml Starflower Oil
Column: C18 5µm, 250x4.6mm
Eluent A: ACN
Eluent B: DCM
Gradient: 30-50% B in 40 mins; 50-90% in 2
mins, hold for 3mins
Flow Rate: 1.0ml/min
Inj Vol: 20µl
Detector: neb=40°C, evap=70°C, gas=1.4 SLM
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Environmental Application:
Characterizing Biodiesel
June 27, 2013
Column: BDS Hypersil Cyano 3µm, 250x4.6 mm
Eluent A: 0.4% Acetic acid in Hexane
Eluent B: 0.4% Acetic acid in Methyl tert-Butyl Ether (MTBE)
Gradient: 100% A, hold 5 mins; 0-80% B in 10 mins,
hold 2 mins; 80-0%B in 3 mins
Flow Rate: 1.0 ml/min
Inj Vol: 20µl
Detector: neb=40°C, evap=80°C, gas=0.8SLM
Page 34
1290 Infinity LC
1290 Infinity Quaternary Pump
1290 2D LC
1290 Infinity with ISET
June 27, 2013
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1290 Infinity Quaternary LC
June 27, 2013
Page 36
Innovation: 1290 Infinity Quaternary Pump Quaternary Pump with Binary Pump-like Performance
June 27, 2013
1290 Infinity Quaternary Pump
• Boost Performance
Highest, binary-like accuracy
and precision for composition
and flow, with exceptionally low
delay volume
• Save Time
Accelerates method transfer
from R+D to QA and from
HPLC to UHPLC
• Reduce Costs
One pump for any applications:
Most universal – at HPLC-like
costs of ownership
Page 37
Power Range
1200bar up to 2mL/min
800bar at 5mL/min
For any kind of analysis
Flow Accuracy and Precision
< 0.07 % RSD or 0.01 min SD
± 1.0 % or 10 µL
High RT precision in
isocratic runs
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 Pump Specifications & Benefits
Composition Range
1-99 %
Wide analytical range
Delay Volume
< 350 µL
For fast quaternary gradients
Page 38
1290 Infinity 2D-LC Solution
June 27, 2013
Page 39
Innovation: 1290 Infinity 2D-LC Solution Square your Separation
June 27, 2013
1290 Infinity 2D-LC Solution
• Boost Performance
New levels of separation power,
detection sensitivity and
precision in 2D-LC
• Save Time
A single ultra high peak
capacity run instead of many
conventional runs – with 1min
method setup!
• Reduce Costs
Special features to reduce
solvent and valve cost, upgrade
paths for existing LC-systems
Page 40
Data Analysis
June 27, 2013
For heart-cutting 2D-LC by Agilent‘s OpenLAB CDS ChemStation ed.
For comprehensive 2D-LC Agilent recommends:
GC Image LCxLC Edition Software from GC Image LLC.
Key features offered:
• Direct Agilent datafile import
• Peak identification, integration, annotation
• Comparative Analysis and Visualization
• Compound-libraries
• MS-data handling
Page 41
Hardware Module-flexibility
June 27, 2013
1. Dimension
2. Dimension
1290 Infinity
Binary Pump
1290 Infinity
Binary Pump
1290 Infinity
Autosampler or 1260
HiP Autosampler
Optional
1260/1290
Infinity
Detector
One or two
1290 Infinity
TCC
Optional
1260/1290
Infinity
Detector
1260/1290
Infinity
detector
1260 Infinity Binary
or Quaternary Pump
1260 Infinity
Capillary Pump 1260 Infinity
Autosampler
For 1st
dimension
chromatogram
and peak-
triggering
To monitor
waste-line
Almost any Agilent pump or autosampler in the 1st dimension!
Almost any detectors are supported!
A 1290 Infinity Binary Pump for the 2nd dimension is required.
Page 42
Applications examples - components of beer samples
June 27, 2013
Separation by RP (C18)
Sepa
ration b
y R
P (
Phe
nyl)
Analysis of two different commercially available beer samples with different
separation modes
1st dimension - Reverse Phase (C18) 2nd dimension - Reverse Phase (Phenyl)
Beer A Beer B
Page 43
1290 Infinity Binary LC w. ISET
Page 44
Innovation: ISET Intelligent System Emulation Technology
June 27, 2013
Intelligent System Emulation Techn.
• Boost Performance
The same retention times
and peak resolution as the
target system
• Save Time
No method redevelopment or
system modification required
• Reduce Costs
Minimized investment and
service costs, with one
system for all methods
Page 45
June 27, 2013
What is Method Transfer? Different Aspects
From particle size to particle size (HPLC to UHPLC)
often associated with a change of instruments due to constraints in max. pressure, extra-
column band broadening etc.. Some fine-tuning of method might be required due to
frictional heating effects
available tools : Agilent Method translator, Third Party Method development SW (ACD
Labs, ChromSword etc.)
From one eluent type or phase chemistry to an other
Method development
available tools: Method development SW (Chromsword, ACD Labs, etc.)
From column dimension to column dimension (e.g. 4.6 to 2.1 mm i.d., 50 mm to 100 mm)
recalculate flow rates, recalculate gradient times, adjust connection capillaries and flow
cells, due to delay volume results may vary, method might need revalidation
available tools: Agilent Method translator
From Instrument to Instrument
with method/instrument change: Isocratic Hold / Pre-injection, Vol. Modification
available tools w/o method or instrument change : none until ISET
Page 46
Intelligent System Emulation Technology Agilent Technical and Application Notes
Public
ation n
um
ber
5990
-9545E
N
Public
ation n
um
ber
5990
-9546E
N
Public
ation n
um
ber
5990-9
703E
N
Public
ation n
um
ber
5990-9
703E
N
Public
ation n
um
ber
5990-9
703E
N
Page 47
Instrument Control Framework
June 27, 2013
Page 48
Innovation: Instrument Control Framework Integrate your Agilent LC with other CDS
June 27, 2013
Instrument Control Framework
• Boost Performance
Full functionality of all Agilent
LC Systems in non-Agilent
data systems
• Save Time
Fast plug-and-play updates
to latest Agilent LC features,
modules, and systems
• Reduce Costs
Access new Agilent features
in non-Agilent software,
without extra coding