Agilent Technologies 1260 Infinity Quaternary LC VL User manual

Agilent Technologies
Agilent 1260 Infinity
Quaternary LC VL
System Manual and
Quick Reference

Agilent 1260 Infinity Quaternary LC VL Manual and Quick Reference
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Manual Part Number
G1311-90310
Edition
06/2010
Printed in Germany
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76337 Waldbronn
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For Research Use Only

Agilent 1260 Infinity Quaternary LC VL Manual and Quick Reference 3
In This Book
In This Book
This book describes the Agilent 1260 Infinity Quaternary LC VL
1 Introduction
This chapter gives an introduction to the Agilent 1260 Infinity Quaternary LC
VL, the underlying concepts and the features of the Agilent 1260 Infinity
Quaternary LC VL.
2 Specifications
This chapter provides information about specifications for the LC system.
3 Optimization of the Agilent 1260 Infinity Quaternary LC VL
This chapter considers how to apply the theory and use the features of the LC
system to develop optimized separations.
4 System Setup and Installation
This chapter includes information on software installation, stack
configurations and preparing the system for operation.
5QuickStartGuide
This chapter provides information on data acquisition and data analysis with
the Agilent 1260 Infinity Quaternary LC VL.
6 Appendix
This chapter provides addition information on safety, legal, web and the Edit
Entire Method.

4 Agilent 1260 Infinity Quaternary LC VL Manual and Quick Reference
Contents
Contents
1 Introduction 7
Introduction to the Agilent 1260 Infinity Quaternary LC VL 8
Features of the Agilent 1260 Infinity Quaternary LC VL 10
System Components 11
Optimizing the Stack Configuration 12
Quaternary pump 17
Autosampler 19
Thermostatted column compartment 24
Detector 25
2 Specifications 37
Physical Specifications 38
Performance Specifications 40
3 Optimization of the Agilent 1260 Infinity Quaternary LC VL 49
Optimizing the Pump 50
Optimizing the Autosampler 54
Optimizing the Thermostatted Column Compartment 61
Optimizing the Detector Regarding to the System 62
Optimizing Detection with DAD 63
Optimizing Detection with VWD 78
4 System Setup and Installation 83
Installing Software 84
Installing the Modules 85
5QuickStartGuide 91
Preparing the System 92
Setting Up the Method 97

Agilent 1260 Infinity Quaternary LC VL Manual and Quick Reference 5
Contents
6 Appendix 99
Safety 100
The Waste Electrical and Electronic Equipment Directive 103
Lithium Batteries Information 104
Radio Interference 105
Sound Emission 106
Solvent Information 107
UV-Radiation 108
Declaration of Conformity for HOX2 Filter 109
Agilent Technologies on Internet 110

6 Agilent 1260 Infinity Quaternary LC VL Manual and Quick Reference
Contents

7
Agilent 1260 Infinity Quaternary LC VL Manual and Quick Reference
Agilent Technologies
1
Introduction
Introduction to the Agilent 1260 Infinity Quaternary LC VL 8
Concept of the Quaternary LC VL System 8
System Properties 8
Features of the Agilent 1260 Infinity Quaternary LC VL 10
System Components 11
Optimizing the Stack Configuration 12
One Stack Configuration 12
Two Stack Configuration 15
Quaternary pump 17
Hydraulic Path 18
Autosampler 19
Sequences 21
Thermostatted column compartment 24
Detector 25
Diode-Array Detector (DAD) 25
Variable Wavelength Detector (VWD) 30
This chapter gives an introduction to the Agilent 1260 Infinity Quaternary LC VL,
the underlying concepts and the features of the Agilent 1260 Infinity
Quaternary LC VL.

8 Agilent 1260 Infinity Quaternary LC VL Manual and Quick Reference
1Introduction
Introduction to the Agilent 1260 Infinity Quaternary LC VL
Introduction to the Agilent 1260 Infinity Quaternary LC VL
Concept of the Quaternary LC VL System
The Agilent 1260 Infinity Quaternary LC VL offers the most flexibility for
solvent selection and automation in HPLC method development, research and
all HPLC applications requiring continuous access to a wide range of solvent
choices. The availability to rapidly switch between methods using different
solvents and the capability of using binary, ternary or quaternary solvent
gradients make the Agilent 1260 Infinity Quaternary LC VL the most flexible
system on the market.
System Properties
The Agilent 1260 Infinity Quaternary LC VL is ideally suited for multi-method,
high-throughput workflows. It offers:
•Convenient access to four solvents for isocratic or gradient analysis for
rapid method development and speed up preparation of mobile phases and
flushing the HPLC system.
•Pressure range up to 400 bar.
•A wide flow range up to 10 mL/min and a delay volume of 800 – 1100 µL
supports narrow-bore, standard and semi-preparative applications.
•Easy programming and control through the Instant Pilot G4208A (requires
firmware B.02.08 or above) (p/n G4208-67001) or through an Agilent Data
System.
•Included micro vacuum degasser offers high degassing efficiency for
trouble-free operation and highest performance and completely eliminates
the need for helium sparging.
•Direct front access for quick exchange of maintenance parts.
•Fast problem identification by self-diagnostics, built-in log books and
preprogrammed test methods.

Agilent 1260 Infinity Quaternary LC VL Manual and Quick Reference 9
Introduction 1
Introduction to the Agilent 1260 Infinity Quaternary LC VL
•Early maintenance feedback (EMF) that continuously tracks long-term
instrument usage and user-defined limits with feedback message when limit
is exceeded.
•Upgradeability and expandability with the complete range of Agilent 1200
Infinity Series HPLC modules.
•Agilent Data System helps you manage your lab for best chromatographic
quality with intuitive diagnostic and monitor capabilities and alert
functions to notify you of problems.

10 Agilent 1260 Infinity Quaternary LC VL Manual and Quick Reference
1Introduction
Features of the Agilent 1260 Infinity Quaternary LC VL
Features of the Agilent 1260 Infinity Quaternary LC VL
The Agilent 1260 Infinity Quaternary LC VL is designed to offer the greatest
flexibility for performing analytical liquid chromatography using all types of
current and emergent column technologies.
The quaternary system as described in this manual offers:
•Gradients of up to 4 different solvents.
•Pressure range up to 400 bar.
•Sophisticated pump control to deliver very low chromatographic noise and
very low acoustic noise for better results and better working environment.
•Degasser and automatic purge valve integrated into pump module.
•Variable volume autosampler with reduced delay volume, reduced
carryover and the option to operate as a fixed loop autosampler.
•Thermostated column compartment with a pressure range up to 600 bar
•Choice of detectors (a set of different flow cells is available for different
detectors to fit application needs regarding flow ranges (nano scale, micro
scale, standard and preparative applications) and pressures):
•Diode-array detector with greatly enhanced sensitivity and baseline
stability using cartridge cell system with optofluidic waveguides (data
collection rate up to 80 Hz with full spectral information) or
•Variable wavelength detector.

Agilent 1260 Infinity Quaternary LC VL Manual and Quick Reference 11
Introduction 1
System Components
System Components
Solvent cabinet
The solvent cabinet is a case to keep four bottles with solvent standing framed
and stable, so that they can be connected by solvent tubings to the LC System.
Quaternary pump
The quaternary pump generates gradients by low pressure mixing from four
individual solvent channels.
Autosampler with/without thermostat
The autosampler is specifically designed for the Agilent 1200 Infinity Series
system for increased analysis speed with sensitivity, resolution, and precision.
Thermostatted column compartment
This stackable temperature-controlled column compartment is used for
heating and cooling in order to meet extreme requirements of retention time
reproducibility.
Diode array or variable wavelength detector
Signals triggered by UV absorption are sampled to be converted to electrical
signals in order for display and software handling.
The Agilent 1260 Infinity Quaternary LC VL are described in more detail in
the following sections. All modules are stackable, see “One Stack
Configuration” on page 12 and “Two Stack Configuration” on page 15.

12 Agilent 1260 Infinity Quaternary LC VL Manual and Quick Reference
1Introduction
Optimizing the Stack Configuration
Optimizing the Stack Configuration
You can ensure optimum performance by installing the system in following
configurations. These configurations optimize the system flow path, ensuring
minimum delay volume.
One Stack Configuration
Ensure optimum performance by installing the modules of the Agilent 1260
Infinity LC System in the following configuration (see Figure 1 on page 13 and
Figure 2 on page 14). This configuration optimizes the flow path for minimum
delay volume and minimizes the bench space required.

Agilent 1260 Infinity Quaternary LC VL Manual and Quick Reference 13
Introduction 1
Optimizing the Stack Configuration
Figure 1 Recommended Stack Configuration (Front View)
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14 Agilent 1260 Infinity Quaternary LC VL Manual and Quick Reference
1Introduction
Optimizing the Stack Configuration
Figure 2 Recommended Stack Configuration (Rear View)
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Agilent 1260 Infinity Quaternary LC VL Manual and Quick Reference 15
Introduction 1
Optimizing the Stack Configuration
Two Stack Configuration
To avoid excessive height of the stack when the autosampler thermostat is
added to the system it is recommended to form two stacks. Some users prefer
the lower height of this arrangement even without the autosampler
thermostat. A slightly longer capillary is required between the pump and
autosampler. See Figure 3 on page 15 and Figure 4 on page 16.
Figure 3 Two stack configuration (front view)
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16 Agilent 1260 Infinity Quaternary LC VL Manual and Quick Reference
1Introduction
Optimizing the Stack Configuration
Figure 4 Two stack configuration (rear view)
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Agilent 1260 Infinity Quaternary LC VL Manual and Quick Reference 17
Introduction 1
Quaternary pump
Quaternary pump
The quaternary pump is based on a two-channel, dual-plunger in-series design
which comprises all essential functions that a solvent delivery system has to
fulfill. Metering of solvent and delivery to the high-pressure side are
performed by one pump assembly which can generate pressure up to 400 bar.
Degassing of the solvents is done in a built-in vacuum degasser. Solvent
compositions are generated on the low-pressure side by a high-speed
proportioning valve (MCGV).
The pump assembly includes a pump head with a passive inlet valve and an
outlet valve. A damping unit is connected between the two plunger chambers.
A purge valve including a PTFE frit is fitted at the pump outlet for convenient
priming of the pump head.
An active seal wash (optional) is available for applications using concentrated
buffers as solvents.

18 Agilent 1260 Infinity Quaternary LC VL Manual and Quick Reference
1Introduction
Quaternary pump
Hydraulic Path
Figure 5 Hydraulic Path of the Quaternary Pump
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Agilent 1260 Infinity Quaternary LC VL Manual and Quick Reference 19
Introduction 1
Autosampler
Autosampler
Figure 6 Overview of the Autosampler
The Agilent 1260 Infinity Autosampler is designed to offer the well-established
Agilent flow-through design with variable volume injection and to achieve
extremely low carryover. The small hydraulic volume of the flow path is suited
to fast gradients and the ability to use overlapped injections and automatic
delay volume reduction (ADVR) contribute to faster cycle times and even
faster gradient delivery to the column. The system draws exactly the set
volume of sample solution without waste and achieves high reproducibility
across the whole range of possible injection volume. The autosampler is
controlled from G4208 A Instant Pilot or from the Agilent Data System.
Three sample-rack sizes are available. The standard full-size rack holds
100 × 1.8 mL vials, while the two half-size racks provide space for 40 × 1.8 mL
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20 Agilent 1260 Infinity Quaternary LC VL Manual and Quick Reference
1Introduction
Autosampler
vials and 15 × 6 mL vials respectively. Any two half-size rack trays can be
installed in the autosamplers simultaneously. A specially designed
sample-rack holding 100 × 1.8 mL vials is available for use with thermostatted
autosamplers. The half-size racks trays are not designed for an optimal heat
transfer when they are used with a thermostatted autosampler.
The autosamplers transport mechanism uses an X-Z-Theta movement to
optimize vial pick-up and return. Vials are picked up by the gripper arm, and
positioned below the sampling unit. The gripper transport mechanism and
sampling unit are driven by motors. Movement is monitored by optical sensors
and optical encoders to ensure correct operation. The metering device is
always flushed after injection to ensure minimum carry-over.
The module uses an analytical head providing injection volumes from
0.1 to 100 µL for pressures up to 600 bar.
The six-port injection valve unit (only 5 ports are used) is driven by a
high-speed hybrid stepper motor. During the sampling sequence, the valve unit
bypasses the autosampler, and directly connects the flow from the pump to the
column. During injection and analysis, the valve unit directs the flow through
the autosampler which ensures that the sample is injected completely into the
column, and that any sample residue is removed from the metering unit and
needle from before the next sampling sequence begins. Different valves are
available.
Control of the vial temperature in the thermostatted autosampler is achieved
using the additional Agilent 1260 Infinity ALS thermostat. Details of this
module are given in the Agilent 1260 Infinity Autosampler Thermostat
manual.