Agilent Technologies InfinityLab LC Series User manual

Agilent Technologies
Agilent InfinityLab LC Series
1260 Infinity II SFC System
User Manual

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G4301-90104
Edition
02/2017
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3
In This Guide...
In This Guide...
This manual covers the Agilent InfinityLab LC Series 1260 Infinity II SFC
System.
1 Introduction to Supercritical Fluid Chromatography (SFC)
This chapter provides an overview of the history, theory and benefits of SFC.
2 Site Requirements and Specifications
This chapter provides information on environmental requirements, physical
and performance specifications only for the Agilent 1260 Infinity II SFC
System.
3 Installing the Agilent 1260 Infinity II SFC System
This chapter provides an overview of the installation and setup of the
hardware and software
4 Configuring the System
How to configure the Agilent 1260 Infinity II SFC System and Agilent 1260
Infinity II SFC Control Module in OpenLAB CDS.
5 Using the Agilent 1260 Infinity II SFC Control Module
This chapter provides information and hints on the use of the SFC System.
6 Maintenance and Repair
In this chapter only the SFC specific procedures are described. For procedures
similar to the Agilent module procedures, please refer to the single module
manuals (G4782A, G4767A, G7116A, G7115/65A)
7PartsforMaintenance
This chapter provides information on parts for maintenance and repair.

4
In This Guide...
8 Identifying Cables
This chapter provides information on cables used with the modules.
9 Appendix
This chapter provides addition information on safety, legal and web.

5
Contents
Contents
1 Introduction to Supercritical Fluid Chromatography (SFC) 7
History of SFC 8
Theory of SFC 9
Benefits of SFC 10
Common flow path overview for packed column SFC instrumentation 11
The Agilent 1260 Infinity II SFC System 12
2 Site Requirements and Specifications 21
Site Requirements 22
Specifications 26
3 Installing the Agilent 1260 Infinity II SFC System 31
Hardware Installation 32
4 Configuring the System 55
Configuring the SFC system in OpenLAB CDS 56
Setting up the Method 60
Status 62
Control 63
5 Using the Agilent 1260 Infinity II SFC Control Module 65
Powering up the Module 66
Power-up Sequence and Operational Control States 67
Operational Control States 68
Controlling the Agilent 1260 Infinity II SFC Control Module through the Agilent
OpenLAB CDS 71
Running a method on the SFC system 73
Shutting Down the SFC System 75

6
Contents
6 Maintenance and Repair 77
Inspection and Preventative Maintenance Intervals 78
General Maintenance procedures 80
7PartsforMaintenance 91
Agilent 1260 Infinity II SFC Control Module Parts 92
Agilent 1260 Infinity II SFC DAD Parts 94
8 Identifying Cables 95
Cable Overview 96
Analog Cables 98
Remote Cables 100
CAN/LAN Cables 104
RS-232 Cables 105
USB 106
9 Appendix 107
General Safety Information 108
The Waste Electrical and Electronic Equipment (WEEE) Directive
(2002-96-EC) 112
Radio Interference 113
Sound Emission 114
Solvent Information 115
Agilent Technologies on Internet 117

7
Agilent Technologies
1
Introduction to Supercritical Fluid
Chromatography (SFC)
History of SFC 8
Theory of SFC 9
Benefits of SFC 10
Common flow path overview for packed column SFC
instrumentation 11
The Agilent 1260 Infinity II SFC System 12
The Agilent 1260 Infinity II SFC Control Module (G4301A) 13
The Agilent 1260 Infinity II SFC Binary Pump (G4782A) 14
The Agilent 1260 Infinity II SFC Multisampler (G4767A) 15
The Agilent 1260 Infinity II Multicolumn Thermostat (G7116A) 16
The Agilent 1260 Infinity II Diode Array Detector WR (G7115A) and
Agilent 1260 Infinity II Multi Wavelength Detector (G7165A) 18
Applications 18
Columns 19
This chapter provides an overview of the history, theory and benefits of SFC.

8
1 Introduction to Supercritical Fluid Chromatography (SFC)
History of SFC
History of SFC
Supercritical fluid chromatography (SFC) was first introduced by Klesper et
al. in 1962 (Klesper, E.; Corwin, A. H.; Turner, D. A. J. Org. Chem. 1962,
27,700.) for the separation and analysis of a porphyrin mixture using open
tubular SFC. The first commercial instruments using packed columns were
available from Hewlett-Packard (HP) in 1982. Since then, several vendors have
developed and commercialized packed column SFC instrumentation for
analytical as well as for preparative separation. SFC is widely accepted for the
separation of chiral compounds and increased user interest has been observed
for a wide spectrum of small to medium sized molecules due to the analysis
speed achieved and the low solvent consumption.
The latest introduction of analytical SFC instrumentation, the Agilent 1260
Infinity II SFC Control Module coupled to an Agilent 1260 Infinity II Binary
LC system optimized for SFC.

9
Introduction to Supercritical Fluid Chromatography (SFC)
1
Theory of SFC
Theory of SFC
Figure 1 State of a solvent
The superior separation properties achieved by SFC can be explained best by
the thermodynamics of liquids and gases (see the phase diagram in Figure 1
on page 9). Above a critical pressure, liquids can no longer enter the gaseous
state; similarly, above a critical temperature, gases cannot be converted to
liquids. Above both the critical pressure and temperature (characterized by
the critical point), solvents are in the supercritical state. Under these
conditions, the mobile phases exhibit gaseous as well as liquid-like properties.
The major advantages of this state related to chromatography are improved
diffusion characteristics and mass transfer and low viscosity, which result
in high separation efficiency and fast separation capability.

10
1 Introduction to Supercritical Fluid Chromatography (SFC)
Benefits of SFC
Benefits of SFC
SFC is widely accepted for the analysis and separation of chiral compounds. In
addition, it gains increasing acceptance as a complementary liquid-based
separation technique to HPLC for high-throughput and high-resolution
analysis of complex mixtures. This is due to the thermodynamic properties of
supercritical fluids, which can be exploited for high throughput and high
efficiency. In addition, the mild thermal conditions also allow for the analysis
of thermally labile compounds. Typically, analysis times and column
re-equilibration are decreased by a factor of 3–5 compared to standard HPLC.
With the increasing costs of organic solvents and the environmental
awareness to minimize toxic waste, production SFC is increasingly accepted
as the “green alternative” to normal phase or reversed phase chromatography,
gaining popularity in method development and UV- and MS-based separation
and purification. A variety of parameters, such as stationary phase selection,
mobile phase composition, modifier type and concentration, column
temperature and system pressure, can be easily manipulated to fulfill
separation requirements by influencing, optimizing and exploiting selectivity
in SFC.

11
Introduction to Supercritical Fluid Chromatography (SFC)
1
Common flow path overview for packed column SFC instrumentation
Common flow path overview for packed column SFC
instrumentation
In commercially available SFC systems, CO
2
is initially pumped in liquid state
and is brought into the supercritical state by heating it above the critical
temperature before it enters the high-pressure area of the LC instrument.
After high-pressure mixing with a modifier, the mobile phase passes into an
injector, where the sample is introduced into the supercritical stream, and
further transported to the separation column. The high pressure of the mobile
phase must be maintained downstream of the detector outlet using a
back-pressure regulator to keep the mobile phase in its supercritical condition
over the complete flow path.

12
1 Introduction to Supercritical Fluid Chromatography (SFC)
The Agilent 1260 Infinity II SFC System
The Agilent 1260 Infinity II SFC System
The Agilent 1260 Infinity II SFC System consists of a binary SFC pump
(G4782A), an SFC Multisampler (4767A), a thermostatted column
compartment (G7116A), and either a DAD (G7115A) or MWD (G7165A)
equipped with a high pressure SFC flow cell and the SFC control module
(G4301A).
The SFC control module, redestills and preconditions the CO
2
by boosting the
pressure to just below the column head pressure, relieving the SFC pump of
any compression requirements. This results in low detector noise and
significantly higher sensitivity. Therefore, the Agilent SFC pump receives
pre-conditioned CO
2
, and acts only as metering device for the mobile phase
flow and to form the gradient with the second pump head by adding the
appropriate amount of modifier solvent.
Downstream of the detector, the mobile phase is redirected back into the SFC
control module to an integrated back-pressure regulator that maintains the
back-pressure over the system.
The Agilent 1260 Infinity II SFC system is completely controlled by Agilent
OpenLAB CDS software. The system diagram is shown below:
Figure 2 Agilent 1260 Infinity II Analytical SFC System
Input "Tee"
BPR
from detector
to Agilent SFC pump
to waste
Output "Tee"

13
Introduction to Supercritical Fluid Chromatography (SFC)
1
The Agilent 1260 Infinity II SFC System
The Agilent 1260 Infinity II SFC Control Module (G4301A)
The Agilent 1260 Infinity II SFC Control Module is responsible for all tasks
connected to pre- and post-conditioning of the mobile phase. In contrast, flow
rate, mobile phase composition, detection, column temperature and data
analysis are controlled by the modules of the Agilent 1260 Infinity II SFC
system in combination with the ChemStation software. This includes metering
the carbon dioxide flow and mixing the modifier into the mobile phase by the
SFC binary pump.
In detail, the Agilent 1260 Infinity II SFC Control Module uses vapor-phase
carbon dioxide, redestills it to the liquid state and boosts its pressure to just
under the column head pressure. Since the CO
2
gas is a very poor solvent,
most contaminants in the carbon dioxide are left in the source, which allows
for the use of inexpensive, beverage-grade CO
2
, unlike in any other
commercially available instrument. The SFC control module further recollects
the effluent from the UV (or other) detector and controls the back-pressure up
to 400 bar over the complete system.

14
1 Introduction to Supercritical Fluid Chromatography (SFC)
The Agilent 1260 Infinity II SFC System
The Agilent 1260 Infinity II SFC Binary Pump (G4782A)
The 1260 Infinity II SFC Binary pump is equipped with passive inlet valves
and with special seals and pistons to allow for CO
2
pumping in channel A
while channel B adds organic modifier for either isocratic or gradient
performance. Pump head B is also equipped with a purge valve to allow for
quick changeover of the organic modifier. In addition, it has an integrated
solvent selection 2-channel degasser and built-in active seal-wash on channel
A for increased uptime
Figure 3 Overview of the pump.
Status indicator
Purge valve
Degasser
Pump head A
Pump head B
Leak drain
Power switch

15
Introduction to Supercritical Fluid Chromatography (SFC)
1
The Agilent 1260 Infinity II SFC System
The Agilent 1260 Infinity II SFC Multisampler (G4767A)
In SFC, the complete solvent flow path needs to be pressurized under all
conditions to avoid expansion of the supercritical fluid. With the Feed
Injection Technology the sample volume is drawn under atmospheric pressure
conditions, pressurized to system pressure, and injected by an ultrafast
syringing process. This enables the injection of flexible sample volumes from
0.1 – 90 μL with highest precision, and excellent linearity over a broad volume
range.
Figure 4 Overview of the Multisampler
Status indicator
Needle assembly
Hotel drawer
Leak drain
Power switch

16
1 Introduction to Supercritical Fluid Chromatography (SFC)
The Agilent 1260 Infinity II SFC System
The Agilent 1260 Infinity II Multicolumn Thermostat (G7116A)
Product Description
The Agilent 1260 Infinity II Multicolumn Thermostat (MCT) allows precise
column thermostatting over a broad temperature range: from cooling down to
10 below ambient temperature to heating up to 85 °C, thus providing high
flexibility for optimized speed and selectivity in standard LC separation.
Ultrahigh pressure valves enable a wide range of applications such as column
selection from 4 columns in a single MCT, sample preparation for analyte
enrichment or matrix removal, alternating column regeneration, etc.
Figure 5 Overview of the Multicolumn Thermostat
Recommendations
The temperature of the mobile phase prior to detection is a critical parameter
for minimizing baseline noise recorded in the detector flow cell. The heat
exchanger on the right side of the column compartment is used to pre- heat
the mobile phase before it enters the column, indirectly heating the column.
The heat exchanger on the left side is used to change the temperature of the
mobile phase to achieve minimum noise. This is of crucial importance, since
the refractive index of carbon dioxide responds up to 50 times stronger than
water- based mobile phases; thus, even small changes in temperature can
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17
Introduction to Supercritical Fluid Chromatography (SFC)
1
The Agilent 1260 Infinity II SFC System
significantly affect noise levels. Temperature changes of mobile phase with the
left side heat exchanger e.g. between 38 °C and 49 °C resulted in a variation in
peak-to-peak noise of over an order of magnitude.
The recommended temperature setting on the left heat exchanger in order to
achieve minimum noise is typically between 37 °C and 40 °C for G7115A or
G7165A detectors.
Figure 6 Recommended temperature setting on the left heat exchanger
37°C and 40°C
for detection with
G7115A and G7165A
To achieve minimum noise
set left heat exchanger
temperature between

18
1 Introduction to Supercritical Fluid Chromatography (SFC)
The Agilent 1260 Infinity II SFC System
The Agilent 1260 Infinity II Diode Array Detector WR (G7115A) and
Agilent 1260 Infinity II Multi Wavelength Detector (G7165A)
The system can be equipped either with a DAD or an MWD using a high
pressure detector flow cell suitable for SFC (10 mm path length, 13 μL
volume), with short transfer tubing to minimize peak broadening. Electronic
temperature control provides highest baseline stability and stable sensitivity
values under fluctuating temperature and humidity conditions. This feature
aids greatly in minimizing detector noise, and now enables impurity and EE
(enantiomeric excess) analysis by SFC, particularly when using elevated
temperatures.
Applications
SFC has gained a wide interest and acceptance in many small molecule
applications because of its high separation speed and efficiency, selectivity,
low operating costs, and due to low generation of organic solvent waste.
Important applications have been developed for the analysis of
pharmaceutical drugs, natural products, fatty acids, vitamins, pesticides,
lipids and chiral compounds. See Figure 7 on page 19.

19
Introduction to Supercritical Fluid Chromatography (SFC)
1
The Agilent 1260 Infinity II SFC System
Columns
In contrast to reversed phase separation, there is no universal stationary
phase available for SFC separations. Most typically used stationary phases are
ethyl pyridine, diol, cyano, amino, Silica and SCX columns. This usually leads
to additional effort to screen different columns in order to achieve optimum
separation. On the other hand, it provides a valuable tool for achieving
different selectivities for a given analyte mixture. A review of column
developments for SFC was recently published by T.Berger, B.Berger &
R.E.Majors in LCGC North America, May 1, 2010
Figure 7 Where SFC Fits In: from non-polar to highly-charged
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20
1 Introduction to Supercritical Fluid Chromatography (SFC)
The Agilent 1260 Infinity II SFC System