Agilent InfinityLab SFC Solutions
EXTEND YOUR APPLICATION REACH WITH MAXIMUM EFFICIENCY
UHPLC SFC
0.5 1.0 1.5 2.00
50
100
150
200 HPLC
SFC0
2
4
6
8
10
2
Unleash the power of SFC. Separate at high flow rates up to 600 bar. Achieve analysis speeds up to 10-times faster than HPLC.
ANALYTICAL EFFICIENCY
Exploit the full orthogonality of supercritical fluid chromatography. Compare SFC and UHPLC results on one system.
INSTRUMENT EFFICIENCY
Reduce costs of solvent purchase and waste disposal by a factor of 5. Eliminate toxic solvents. Make your lab leaner and greener.
LABORATORY EFFICIENCY
INTRODUCTION
EXTEND YOUR APPLICATION REACH WITH MAXIMUM EFFICIENCY
Agilent’s new supercritical CO2-based technologies deliver robust, reliable, and reproducible separation results for diverse sample types with the widest selectivity from polar to nonpolar. What’s more, Agilent InfinityLab SFC Solutions maximize efficiency, use less solvents, and lower costs while being fully compatible with the full range of Agilent mass spectrometers.
3
COMPLETE SOLUTIONSAgilent InfinityLab SFC Solutions are truly complete end-to-end solutions, combining superior instruments and columns, smart supplies, intuitive software, and dedicated services.
Services and Support
Learn how to gain the maximum benefit from your InfinityLab SFC Solutions through our comprehensive suite of services and learning solutions.
Software and Informatics
Stay in full control of all your SFC applications through OpenLAB CDS software, and optimize development of your SFC methods with Method Scouting Wizard.
InfinityLab Instruments, Columns and Supplies
Designed to work together in perfect harmony, Agilent InfinityLab SFC instrumentation and single quadrupole mass selective detectors combine with Agilent columns and InfinityLab supplies to maximize the efficiency of your SFC workflows.
UHPLC SFC
4
EXPERIENCE ROBUST PERFORMANCE FOR HIGHEST CONFIDENCE IN YOUR DAILY RESULTS
TECHNOLOGY
InfinityLab SFC Solutions are fully-fledged members of the InfinityLab family. While some modules have been reengineered to cater for the demands of liquefied CO2 as mobile phase, many modules remain unchanged and are common to both SFC and UHPLC systems.
SUPERIOR SOLVENT DELIVERYHighest flow accuracy and precision up to 5 mL/min at 600 bar. Improved usability and extended lifetime through integrated degasser, solvent selection, valve and active seal wash.
EFFICIENT COLUMN HANDLINGHands-free selection of up to four columns. Independent heating zones for optimum separation and postcolumn conditioning.
HYBRID, MULTIPURPOSE SAMPLINGNew, unique feed injection for SFC sampling. Classic flow-through injection for UHPLC sampling. Flexible choice of sample containers such as 2 or 6 mL vials, or microtiter plates. Highest capacity up to 432 samples in 2 mL vials.
SFC feed injection
• Extended injection volume range from 0.1 to 90 µL
• Full choice of sample transfer solvent and injection speed
• No delay volume
• Reduced sample solvent effects
Agilent 1260 Infinity II SFC/UHPLC Hybrid SystemSwitch between SFC and UHPLC with a simple mouse click! With the easy-to-install addition of a 2-position/10-port valve and a 1260 Infinity II LC Pump, your system becomes a fully operational hybrid SFC/UHPLC solution. The option of running both SFC and UHPLC applications on the same system allows you to maximize the utilization of your analytical equipment. The core of the hybrid solution is the 1260 Infinity II SFC Multisampler. With feed injection for SFC sampling and classic flow-through injection for UHPLC sampling, the SFC Multisampler is flushed automatically when switching between techniques.
5
EXTENDED APPLICATION REACHCheck out our broad range of detection options, see page 10.
CONVENIENT CO2 HANDLINGProgrammable backpressure gradients for shorter analysis times. Full-flow transfer to MS or ELSD with new low-dispersion nozzle. Increased robustness and extended lifetime through major overhaul of booster pump.
PERFECTLY MATCHED SUPPLIESSmaller footprint, easy-to-grip solvent bottles with Stay Safe caps make solvent handling more convenient – just two of the many InfinityLab LC supplies that help deliver reliable, robust performance.
15 32
6
DEVELOP YOUR SFC METHODS WITH MAXIMUM EFFICIENCY
TECHNOLOGY
InfinityLab Method Development Solutions comprise instruments, columns, software, and services that work together in perfect harmony, helping you to develop your chromatographic methods with maximum efficiency.
Dedicated hardware for the highest degree of automationThe flexibility of the InfinityLab SFC solutions helps you to solve your toughest development challenges. Simply install an external solvent-selection valve and gain immediate, automated access to 15 different solvents. Cluster two, three, or even four 1290 Infinity II Multicolumn Thermostats for automated screening of up to 32 columns.
Columns and supplies for simple and quick method developmentZORBAX and InfinityLab Poroshell columns are manufactured end-to-end by Agilent and monitored with multiple QC tests, so you can be assured of rugged, reproducible performance and long column lifetimes. From Quick Connect capillaries through Stay Safe caps, our wide range of parts, kits, and accessories are designed, manufactured, and tested under a quality system registered to ISO 9001.
Download our Quick Reference Guide for SFC Supplies from www.agilent.com, search for 5991-8116EN.
Easy, automated access to columns and solvents gives you ultimate flexibility in method development.
7
Dedicated software accelerates method developmentAgilent OpenLAB CDS ChemStation Edition provides the versatility you need to develop your SFC methods. Agilent Method Scouting Wizard helps you to generate methods sequences for complex screening campaigns within minutes. Agilent MassHunter Walkup software allows users of different skill levels to perform their own SFC and SFC/MS analyses. Results show up in the submitter’s in-box automatically.
Method Scouting Wizard is an easy-to-use but highly effective tool to develop SFC and UHPLC methods with less effort. Use this add-on for OpenLAB CDS ChemStation to reduce time spent on manual scanning of different parameters such as mobile phases and columns.
OpenLAB CDS 2 – One software for Agilent SFC, LC, LC/MS, GC, GC/MS and other vendors’ instruments provides the same user experience and enables viewing data across techniques and instruments with the same user interface.
The right tools for the jobWhether you run a small lab with few instruments or a larger operation with a requirement for networked solutions, OpenLAB software products cover all demands. OpenLAB CDS 2 increases productivity in analytical QC labs and gives you peace of mind when regulatory agencies audit or inspect your operation. With support for SFC and single quadrupole mass selective detection, OpenLAB CDS 2 helps you to:• Deliver results quickly and securely by performing calculations and
producing reports in the protected environment of the software
• Identify nonconforming samples quickly with Peak Explorer visualization tool and MatchCompare add-on
Advanced tools for advanced applicationsWhen combining your SFC solution with triple quadrupole or quadrupole time-of-flight mass spectrometers, Agilent MassHunter software is designed to make your MS analyses faster, easier, and more productive. MassHunter’s modular architecture lets you integrate new features to match your workflow requirements.
Normal-phase HPLC HILIC
Reversed-phase HPLC Ion chromatography
Ion pairing
SFC: CO2 and organic modifiers SFC: CO2, modifiers, additives, and water
SFC: CO2, modifiers and additives
Hydrocarbons Aldehydes and ketones Hydroxy acids and poly acids Peptides
Fatty acids and waxes Analine and benzoic acids
Zwitter ions
Primary amines
Ions
Proteins
Normal-phase HPLC HILIC
Reversed-phase HPLC Ion chromatography
Ion pairing
SFC: CO2 and organic modifiers SFC: CO2, modifiers, additives, and water
SFC: CO2, modifiers and additives
INCREASING POLARITY
DNA/RNAEthers and esters
SFC: pure CO2
8
SWITCH TO SFC AND PROTECT YOUR INVESTMENT AND THE ENVIRONMENT
Switch to SFC now and benefit from an immediate reduction in the costs of solvent purchase and waste disposal. What’s more, the virtual elimination of hazardous solvents makes your lab a cleaner place to work as well as protecting the environment.
Except for extremely polar separation techniques such as ion exchange chromatography, SFC largely covers the same application areas as normal-phase LC, reversed-phase LC, and hydrophilic interaction chromatography (HILIC). The applicability of SFC ranges from nonpolar hydrocarbons through high-polarity amines, hydroxy acids, and peptides.
SFC AT A GLANCE
Leaner and greener – but where can I use SFC?As a fully orthogonal separation technology to liquid chromatography, SFC has the potential to replace LC in a huge number of applications that span a wide range of analyte polarities. An impressive number of publications on completely diverse applications report SFC as the superior choice to LC. The main area where SFC delivers better results covers applications currently using normal-phase LC such as the analysis of chiral molecules.
Hept
ane
NPLC
GHS classification
RPLC SFC
Acet
onitr
ile
(+ m
odifi
er)
Hept
ane
Acet
onitr
ile
CO2
NPLC
Cost per liter
RPLC SFC
100 %
5 - 95
%
5 - 40
%
NPLC
Proportion of organic solvents
RPLC SFC
P [bar]
250 300 350 4001200
10
100
1,000
10,000
triple point
critical point
supercriticalfluid
liquid
solid
gas
T [K]
9
Reduce hazardous solventsReplacing normal-phase LC by SFC means a significant reduction in the need for hazardous solvents such as acetonitrile, dichloromethane, or heptane. Your lab becomes a greener place to work where your only concern is the GHS classification of typical SFC modifiers such as methanol, ethanol, or isopropanol.
Reduce solvent costsInfinityLab SFC Solutions use food-grade carbon dioxide as the primary mobile phase. Food-grade CO2 can be purchased at a fraction of the cost of the primary solvents used in other modes of liquid chromatography. Significant savings are also made in waste disposal with only small amounts of modifiers being used.
Reduce waste productionThe proportion of organic modifier used in SFC gradients is generally significantly lower than in normal- or reversed-phase LC. As a result, SFC generates far less waste. The major solvent, carbon dioxide, simply evaporates and diffuses safely into the atmosphere.
A supercritical fluid is any substance at a temperature and pressure above its critical point, where distinct liquid and gas phases do not exist. Close to the critical point, small changes in pressure or temperature result in large changes in density. These characteristics make it possible to adjust many physical properties so that a supercritical fluid can be used as a substitute for organic solvents in industrial and laboratory processes such as chromatography.
In supercritical fluid chromatography, carbon dioxide is the most commonly used solvent whereby cosolvents are added to modify the mobile phase polarity and thereby change the selectivity of the separation system.
Benefits at a Glance
SFC
Full flow
ELSD/MSD
10
MATCH DETECTION TECHNIQUE TO YOUR APPLICATION NEEDS
APPLICATIONS
InfinityLab SFC Solutions extend and broaden your application reach by providing an orthogonal separation mechanism that can be combined with a wide variety of different detection techniques. Choose a detector that matches your needs for selectivity and sensitivity, the number of analytes, and the complexity of your sample matrix.
Evaporative light scattering detection 1290 Infinity II ELSD Quantitative and qualitative analysis of nonvolatile analytes that lack UV-visible chromophores such as sugars.
Flame ionization detection* 1260 Infinity II SFC Sensitive analysis of fuels by flame ionization, according to standard methods such as ASTM D5186. *SIM Scientific Instruments Manufacturer GmbH
UV-visible detection 1260 Infinity II DAD Determination of enantiomeric ratios or quantification of a small number of known analytes.
Full-flow transfer for highest sensitivityTo achieve highest sensitivity, transfer the complete SFC effluent containing your precious sample to the ELSD, MSD, or LC/MS. The new low-dispersion nozzle significantly reduces the adverse effects of passing through the backpressure regulator.
SFC
Split flow
MS
11
Mass selective detection InfinityLab MSD Simplified peak tracking in method development or quantification of a few analytes in simple matrixes.
Triple quadrupole mass spectrometry 6470 Triple Quadrupole LC/MS Highly sensitive forensic screening, confirmation, and quantification of drugs of abuse, pesticides, or vitamins in challenging matrixes.
Quadrupole time-of-flight mass spectrometry 6545 Quadrupole Time-of-Flight LC/MS Full screening of suspect analytes or unknowns in complex matrixes using data-mining tools, and personal compound databases and libraries (PCDL).
Flow splitting for highest resolutionTo achieve highest resolution, use a flow splitter in front of the backpressure regulator so that only a portion of the SFC eluent is transferred to the detector. This setup keeps the dispersion volume to a minimum and reduces band broadening.
Number of analytes
2.5 5 7.5 10 12.5 15 17.5 20 22.5
0
20
40
60
80
100
120
PGTB
QH
TROL
OX BHA
OG
LG
δ-TTγ-TTα-TTδ-TPγ-TPα-TP
BHT
min
mAU
2.5 5 7.5 10 12.5 15
0
10
20
30
40
50
60
70
80TB
QHTR
OLOX
BHT
BHA
mAU
min
α-TTβ-TPγ-TPδ-TPγ-TT
δ-TTα-
TP
UHPLC SFC
12
InfinityLab SFC Solutions allow you to separate and quantify complex mixtures faster and more efficiently than ever before. Deploying SFC in your lab gives you a technique that is orthogonal to normal- or reversed-phase LC and enables you to reach beyond your current application spectrum.
EXTEND YOUR APPLICATION REACH WITH STATE-OF-THE-ART SFC
APPLICATIONS
Separation and UV detection of 14 antioxidants found in vegetable oils by UHPLC (left) and by SFC (right). The concentration of each antioxidant was 10 μg/mL.
Use SFC to separate your most challenging samples
Vitamin E tocopherols and tocotrienols have different biological activities and chemical properties so it is important to identify and quantify each vitamer separately. SFC enables complete resolution of all vitamers within a significantly shorter analysis time.
Download Application Note from www.agilent.com, search for 5991-1546EN.
min0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 2.25
mAU
0255075
100125150175200
DAD1 A, Sig=280,4 Ref=360,100
5CB
1.3
85
7CB
1.4
688O
CB 1
.573
5CT
1.7
53
H3CC N
4-cyano-4'-n-pentyl-biphenly (5CB) CAS number: 40817-08-1
4-cyano-4'-n-heptyl-biphenyl (7CB)CAS number: 41122-71-8
4-cyano-4'-n-oxyoctyl-biphenyl (8OCB) CAS number: 52364-73-5
4-cyano-4''-n-pentyl-terphenyl (5CT)CAS number: 54211-46-0
O
H3C
H3C
H3C
C N
C N
C N
min3.2 3.4 3.6 3.8 4.0 4.2 4.4 4.6 4.8
mAU
0
50
100
150
200
25080 µL, 70 µL, 60 µL50 µL40 µL30 µL
20 µL
10 µL
Inj. volume (µL) RT (min) Area Height Width (min) Symm.10 4.337 187.27 79.89 0.036 0.87520 4.336 375.48 148.32 0.039 0.87630 4.331 566.40 197.70 0.045 0.87240 4.326 760.59 231.65 0.053 0.88150 4.320 938.66 246.14 0.062 0.89160 4.314 1,127.29 255.96 0.073 0.90570 4.309 1,315.80 261.58 0.084 0.93380 4.302 1,494.58 261.62 0.096 0.947
0
0.05
0.10
0.15
0.20
0.25
0.30
10 20 30 40 50 60 70 80
Peak
are
a RS
D (%
)
Injection volume versus peak area RSD
Injection volume (µL)
y = 18.706x + 3.9655R2 = 0.9999
0200400600800
1,0001,2001,4001,600
10 20 30 40 50 60 70 80
Peak
are
a
Injection volume (µL)
Linearity of peak area from large volume injection
min3.2 3.4 3.6 3.8 4.0 4.2 4.4 4.6 4.8
mAU
0
50
100
150
200
25080 µL, 70 µL, 60 µL50 µL40 µL30 µL
20 µL
10 µL
Inj. volume (µL) RT (min) Area Height Width (min) Symm.10 4.337 187.27 79.89 0.036 0.87520 4.336 375.48 148.32 0.039 0.87630 4.331 566.40 197.70 0.045 0.87240 4.326 760.59 231.65 0.053 0.88150 4.320 938.66 246.14 0.062 0.89160 4.314 1,127.29 255.96 0.073 0.90570 4.309 1,315.80 261.58 0.084 0.93380 4.302 1,494.58 261.62 0.096 0.947
0
0.05
0.10
0.15
0.20
0.25
0.30
10 20 30 40 50 60 70 80
Peak
are
a RS
D (%
)Injection volume versus peak area RSD
Injection volume (µL)
y = 18.706x + 3.9655R2 = 0.9999
0200400600800
1,0001,2001,4001,600
10 20 30 40 50 60 70 80
Peak
are
a
Injection volume (µL)
Linearity of peak area from large volume injection
13
Solve separation problems faster and more reliably with SFC
The E7 liquid crystal mixture can be separated and quantified within two minutes using SFC with UV detection. This is up to 40-times faster than analysis based on normal-phase LC. Further, SFC delivers improved peak shape for more reliable quantification.
Download Application Note from www.agilent.com, search for 5991-6436EN.
Separation of a mixture of liquid crystal compounds 5CB, 7CB, 8OCB, and 5CT (50 μg/mL, each) within a short run time using a fast gradient.
Feed injection delivers unprecedented area linearity over a wide volume range from 0.1 to 90 μL.
Benefit from unmatched injection precision with flexible injection volumes
The new feed-injection principle achieves unmatched precision and linearity with negligible carryover. It provides full flexibility in the selection of injection volume and eliminates the disadvantages associated with the widely used fixed-loop injection technique.
Download Technical Overview from www.agilent.com, search for 5991-7623EN.
min2.0 2.5 3.0 3.5 4.0
mAU
0
50
100
150
200
250
3003.085
3.561
min2 4 6 8 10
mAU
0
50
100
150
200
250
4.495
5.440 5.731
7.087 7.855
9.928
Separation of propranolol enantiomersunder isocratic conditions:
30 % MeOH25 % MeOH20 % MeOH
Chiral column: IB
14
The analysis speed and the flexibility to handle large and small injection volumes make InfinityLab SFC Solutions the perfect tool for method and compound screening. What‘s more, our SFC systems combine seamlessly with high-end mass spectrometry.
SFC – THE PERFECT TOOL FOR METHOD AND COMPOUND SCREENING
APPLICATIONS
SFC is a fast approach to identify speed-optimized methods for separation of racemates such as propranolol. The final optimized method is shown right.
Develop and optimize new SFC methods for your chiral separations
The InfinityLab SFC Method Development Solution with Agilent Method Scouting Wizard facilitates software-aided method scouting of four chiral columns to develop and optimize new methods.
Download Application Note from www.agilent.com, search for 5991-7624EN.
For Research Use Only. Not for use in diagnostic procedures.
×105
0
1.0
2.0
3.0
4.0
5.0
6.02.957
Acquisition time (min)0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 2.4 2.6 2.8 3.0 3.2 3.4 3.6 3.8 4.0 4.2 4.4 4.6
Coun
ts
×106
1 2 3 4 5 6 7 8
6.856
9 10 11 12 13 14 15 16 17 180
0.5
1.0
1.5
2.0
2.5
3.0
Acquisition time (min)
Coun
ts
15
Determine drugs of abuse at trace levels using SFC with triple quadrupole MS
SFC is an ideal separation technique for fast, quantitative screening of different classes of drugs, ranging from simple amines to complex morphine-like structures such as those analyzed in forensic toxicology applications. Highest analytical sensitivity is achieved by combining SFC with triple quadrupole mass spectrometry.
Download Application Note from www.agilent.com, search for 5991-6747EN.
For Forensic Use.
SFC separation and DMRM detection of 25 drugs within 5 minutes. The limit of detection (LOD) was 30 pg/mL.
Determination of 223 pesticides by DMRM. The first 195 compounds eluted within 10 minutes.
Identify and quantify more than 200 pesticides in a single SFC/MS run
The combined performance of SFC and triple quadrupole MS makes it possible to identify and quantify over 200 pesticides in a single run. Using dynamic multiple reaction monitoring (DMRM) significantly improves the sensitivity of the analysis.
Download Application Note from www.agilent.com, search for 5991-6151EN.
Learn morewww.agilent.com/chem/sfc
Find an Agilent customer centerwww.agilent.com/chem/contactus
USA and Canada1-800-227-9770
Asia [email protected]
This information is subject to change without notice.
© Agilent Technologies, Inc. 2017 Published in the USA, October 15, 20175991-8326EN