1. Manuals
  2. Brands
  3. Agilent Technologies Manuals
  4. Laboratory Equipment
  5. 1290 Infinity LC System
  6. System manual and quick reference

Agilent Technologies 1290 Infinity LC System System Manual And Quick Reference

Hide thumbs
Table of Contents

Advertisement

Quick Links

Download this manual
System Manual and
Quick Reference
Agilent 1290 Infinity
LC System
Agilent Technologies

Advertisement

Table of Contents
loading

Related Manuals for Agilent Technologies 1290 Infinity LC System

Summary of Contents for Agilent Technologies 1290 Infinity LC System

  • Page 1 Agilent 1290 Infinity LC System System Manual and Quick Reference Agilent Technologies...
  • Page 2 1987) or any equivalent agency regulation or contract clause. Use, duplication or dis- closure of Software is subject to Agilent Technologies’ standard commercial license terms, and non-DOD Departments and Agencies of the U.S. Government will Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 3 This manual covers the Agilent 1290 Infinity LC System. 1 Introduction to Ultra-High Performance Liquid Chromatography This chapter gives an introduction to the Agilent 1290 Infinity LC System and the underlying concepts. 2 The Agilent 1290 Infinity LC System - Product Description This chapter discusses the features of the 1290 Infinity LC System.

  • Page 4: Table Of Contents

    Theory of Using Smaller Particles in Liquid Chromatography Benefits of Columns Packed With Sub-2-micron Particles Frictional Heating 2 The Agilent 1290 Infinity LC System - Product Description New Features of the Agilent 1290 Infinity LC System System Components 3 Optimization of the Agilent 1290 Infinity LC System...
  • Page 5 Contents 6 Appendix Safety Information Solvent Information Agilent Technologies on Internet Setting Up a Method using Edit Entire Method Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 6 Contents Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 7: Introduction To Ultra-High Performance Liquid Chromatography

    Introduction to Ultra-High Performance Liquid Chromatography Theory of Using Smaller Particles in Liquid Chromatography Benefits of Columns Packed With Sub-2-micron Particles Frictional Heating This chapter gives an introduction to the Agilent 1290 Infinity LC System and the underlying concepts. Agilent Technologies...

  • Page 8: Theory Of Using Smaller Particles In Liquid Chromatography

    600 bar pressure. In 2009 the range was extended to include RRHD columns capable of routine use at 1200 bar pressure to support the introduction of the Agilent 1290 Infinity LC System with its extensive operating power range up to 1200 bar pressure and 5 ml/min flow rate.
  • Page 9 This type of plot is known as a Van Deemter Curve and is used to determine the optimum flow rate (minimum point of the curve) for best efficiency of separation for a column. Figure 1 A hypothetical Van Deemter curve Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 10 (the slope of the line is reduced). This means that the separation on smaller particles can be further accelerated by increasing the flow rates without significantly reducing efficiency. Figure 2 Van Deemter curve for different particle sizes Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 11 Figure 3 Selecting optimal conditions for HPLC Resolution can be described as a function of three parameters: • column efficiency or theoretical plates (N), • selectivity (), • retention factor (k). Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 12 No matter whether the rapid resolution separation method is being newly developed or simply transferred from an existing conventional method, it is clearly beneficial to have a wide choice of stationary phase chemistries available in a range of column formats. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 13 ZORBAX columns with 3.5 and 5 µm particles. As a result, for any particular ZORBAX phase, the 5.0, 3.5 and 1.8 µm particles provide identical selectivity, which allows easy, fast and secure bidirectional method transfer between conventional LC, UHPLC and preparative LC. Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 14: Benefits Of Columns Packed With Sub-2-Micron Particles

    The Agilent cost savings calculator provides an easy way to calculate the cost savings by switching from conventional HPLC to UHPLC using 1.8 µm particle size columns. This calculator is available on the Agilent Technologies web site along with a method translator calculator – www.chem.agilent.com. The results are presented graphically and in tabular form.
  • Page 15 Introduction to Ultra-High Performance Liquid Chromatography Benefits of Columns Packed With Sub-2-micron Particles Figure 6 Cost savings calculator Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 16 Introduction to Ultra-High Performance Liquid Chromatography Benefits of Columns Packed With Sub-2-micron Particles Figure 7 Relation between particle size, efficiency and analysis time Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 17 Figure 8 Peak capacities of more than 700 can be achieved using a ZORBAX RRHT SB-C18 column (2.1 x 150 mm, 1.8 µm) to analyze a tryptic digest of BSA Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 18: Frictional Heating

    (Figure 9 on page 19 below). The pressure increased to 1070 bar which also indicates that the in-column temperature was lower. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 19 In summary, the use of sub-two-micron packing material offers benefits of increased efficiency, higher resolution and faster separations. The Agilent 1290 Infinity LC System and RRHD columns increase the separation space available and enable more of these benefits to be accessed. The features of the 1290 Infinity LC are discussed in “The Agilent 1290 Infinity LC System -...
  • Page 20 Introduction to Ultra-High Performance Liquid Chromatography Frictional Heating Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 21: The Agilent 1290 Infinity Lc System - Product Description

    Agilent 1290 Infinity LC System Manual and Quick Reference The Agilent 1290 Infinity LC System - Product Description New Features of the Agilent 1290 Infinity LC System System Components This chapter discusses the features of the 1290 Infinity LC System. Agilent Technologies...

  • Page 22: New Features Of The Agilent 1290 Infinity Lc System

    New Features of the Agilent 1290 Infinity LC System New Features of the Agilent 1290 Infinity LC System The Agilent 1290 Infinity LC System is designed to offer the greatest flexibility for performing analytical liquid chromatography using all types of current and emergent column technologies.
  • Page 23 The Agilent 1290 Infinity LC System - Product Description New Features of the Agilent 1290 Infinity LC System • Data collection rates up to 160 Hz with full spectral information. • New range of ZORBAX RRHD sub-two micron particle size columns for operation at pressures up to 1200 bar.
  • Page 24 The Agilent 1290 Infinity LC System - Product Description New Features of the Agilent 1290 Infinity LC System The pressure range offers the ability to work with the latest sub-two micron particles in long columns for high resolution and short columns for fast separation at increased flow rates.

  • Page 25: System Components

    The Agilent 1290 Infinity LC System - Product Description System Components System Components The Agilent 1290 Infinity Binary Pump The Agilent 1290 Infinity Binary Pump contains new technology to overcome the problems of pumping LC solvents at ultra-high pressure and high flow rates: heavy duty drive motors on the pistons;...
  • Page 26 The Agilent 1290 Infinity LC System - Product Description System Components Figure 12 Parts identification and schematic of the 1290 Infinity Binary Pump Each pump head is a dual piston in-series design utilizing novel firmware control and novel piston material, silicon carbide, which efficiently removes heat from the pump.
  • Page 27 The Agilent 1290 Infinity LC System - Product Description System Components chromatographic performance these features makes the pump very quiet in operation. When concentrated buffer solutions are used as mobile phase, the active seal wash option is available for use to extend the life of the pump seals.
  • Page 28 The Agilent 1290 Infinity LC System - Product Description System Components The flow path of the pump has been optimized for minimal delay of gradients and incorporates an innovative mixing system using microfluidics technology. The mixing device, known as the Jet Weaver, employs a network of multi-layer microfluidic channels (120 µm x 120 µm) to unsure the flow is thoroughly...
  • Page 29 The Agilent 1290 Infinity LC System - Product Description System Components The 1290 Infinity Quaternary Pump In contrast, the 1290 Infinity Quaternary Pump is equipped with only one pump head and an additional multi-channel gradient valve (MCGV) to portion the eluents according to the programmed gradient. According to this low pressure mixing principle, the solvents meet in the Inlet Weaver and are therefore already mixed before and in the pump head.
  • Page 30 The Agilent 1290 Infinity LC System - Product Description System Components Table 2 Valve functionalities Pump Pump Agilent Agilent Jet Weaver Jet Weaver Sampler Sampler Waste Waste Filter Filter Figure 14 Standard application Figure 15 Extra-mixing volume setup Pump Pump...
  • Page 31 The Agilent 1290 Infinity LC System - Product Description System Components The standard application (1) is used for the majority of analysis, while the extra-mixing volume setup (2) is applied for any kind of baseline-criticial application where the mixing performance and the dependant UV-baseline can be significantly enhanced by the use of the Agilent Jet Weaver mixer.
  • Page 32 The Agilent 1290 Infinity LC System - Product Description System Components The Agilent 1290 Infinity Autosampler The Agilent 1290 Infinity Autosampler offers the well-established Agilent flow-through design with variable volume injection and takes it to new levels of performance. New inert materials in the metering device seal and needle seat help to achieve extremely low carryover.
  • Page 33 The Agilent 1290 Infinity LC System - Product Description System Components A totally new optional add-on module, the Flexible Cube, works seamlessly with the autosampler to provide additional capabilities. With the addition of the new Flexible Cube module, which comprises a 500 µl syringe pump, one low pressure valve and two high pressure switching valves further options become possible.
  • Page 34 The Agilent 1290 Infinity LC System - Product Description System Components The Agilent 1290 Infinity Thermostatted Column Compartment The Agilent 1290 Infinity Thermostatted Column Compartment (TCC) controls the temperature between 10 °C below ambient and up to 100 °C at 2.5 ml/min and 80 °C at up to 5 ml/min, respectively.
  • Page 35 The Agilent 1290 Infinity LC System - Product Description System Components Figure 19 Quick change valve in TCC Up to three TCC can be “clustered” to allow advanced applications such as switching between eight columns for automated method development or to make additional columns available for different applications.
  • Page 36 The Agilent 1290 Infinity LC System - Product Description System Components The 1290 Infinity Diode-Array Detector The 1290 Infinity Diode-Array Detector is a new optical design using a cartridge cell with optofluidic waveguide technology offering high sensitivity with low dispersion, a wide linear range and a very stable baseline for standard or ultra-fast LC applications.
  • Page 37 The Agilent 1290 Infinity LC System - Product Description System Components The module also incorporates electronic temperature control to further enhance the resistance to ambient temperature effects. Although the hydraulic volume of the Max-Light cartridge cell is very small, the path length is a standard 10 mm.
  • Page 38 The Agilent 1290 Infinity LC System - Product Description System Components Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 39: Optimization Of The Agilent 1290 Infinity Lc System

    Agilent 1290 Infinity LC System Manual and Quick Reference Optimization of the Agilent 1290 Infinity LC System Delay Volume and Extra-Column Volume Delay Volume Extra-Column Volume How to Configure the Optimum Delay Volume How to Achieve Higher Injection Volumes How to Achieve High Throughput...

  • Page 40: Delay Volume And Extra-Column Volume

    With sub-two µm packings the optimum flow rate (from the Van Deemter Curve) is a little higher and so fast chromatography can use three to five times these flow rates yielding delay Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 41: Extra-Column Volume

    This is achieved with the Agilent 1290 Infinity LC System due to the low delay volume of the pump flow path, low volume of the Jet Weaver mixer and low-volume of the flow path through the autosampler.

  • Page 42: How To Configure The Optimum Delay Volume

    Table 4 on page 43 show the component volumes which contribute to system delay volume in the Agilent 1290 Infinity LC System. In the standard configuration with the Agilent 1290 Infinity Binary Pump, with the Jet Weaver mixer, the 1290 Infinity Autosampler and Thermostatted Column Compartment the system delay volume is about 125 µl.
  • Page 43 0.14 0.21 0.30 0.90 1.08 1.28 1.45 0.03 0.09 0.14 0.20 0.60 0.72 0.85 0.97 0.03 0.07 0.11 0.15 0.45 0.54 0.64 0.73 0.02 0.06 0.09 0.12 0.36 0.43 0.51 0.58 Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 44 (see Figure 18 page 32). To make an injection the valve switches from mainpass to bypass so that the metering device can draw the sample into the needle capillary. The Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 45 Max-Light cartridge cell has a low dispersion volume ( volume 1.0 µl) and no further volume optimization is required. In situations where the alternative Agilent Max-Light high sensitivity cell is used to get higher sensitivity the cell Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 46 “Replacing the Jet Weaver in the 1290 Infinity Binary Pump” page 48. Due to the different setup and mixing principle in the Agilent 1290 Infinity Quaternary Pump, the physical delay volume is much bigger and in standard Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 47 Quaternary Pump. The procedure for installing a Jet Weaver in the 1290 Infinity Quaternary Pump is illustrated in “Installing the V380 Jet Weaver in the 1290 Infinity Quaternary Pump” on page 49. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 48 (35 / 100 µl) that are 0.17 mm i.d.). The outlet at the top is connected to the optimized for a low delay volume or best mixing autosampler. performance. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 49 (2). Insert the Jet Weaver to the opening in the front panel (1) Mount both capillary connections to the Jet Weaver and push it down (2). observing the correct orientation. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 50 How to Configure the Optimum Delay Volume Connect the inlet capillary of the Jet Weaver to port 2 of the Multi Purpose Valve. Connect the outlet capillary to port 1. Port 1 Port 2 Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 51: How To Achieve Higher Injection Volumes

    (see Table 7 on page 44) – in a gradient separation injection volumes of about 5 % of the column volume might be achieved whilst maintaining good resolution and peak dispersion. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 52 Figure 21 on page 52. The valve can be conveniently located in the Thermostatted Column Compartment. Figure 21 Sample enrichment Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 53: How To Achieve High Throughput

    For this reason the data processing can be taken off-line to allow the system to focus on running the samples and collecting data. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 54 1290 Infinity pumps. For instance a 1200 Series isocratic pump would be sufficient to perform this task. The setup is illustrated in Figure 22 on page 55. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 55 Optimization of the Agilent 1290 Infinity LC System How to Achieve High Throughput Figure 22 Alternating Column Regeneration Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 56: How To Achieve Higher Resolution

    This section considers how to get higher resolution with any chosen stationary and mobile phases. If an automated method development system was used in Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 57 This is the reason that the 1290 Infinity LC system was designed to go to 1200 bar so that it can run sub-two-micron particles and column length can be increased to 100 mm or 150 mm.
  • Page 58 Any reduction in extra-column volume will reduce dispersion and give better resolution. This is already optimized in the 1290 Infinity LC system with narrow bore (0.12 mm i.d.) capillaries (check that the shortest length is used between column and detector) and the Max-light cartridge flow cell.

  • Page 59: How To Achieve Higher Sensitivity

    4.7 times due to the decreased dilution in the column. For a mass spectrometer detector, the lower flow rates of narrow columns can result in higher ionization efficiencies and therefore higher sensitivity. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 60 • The wavelength indicates the center of the detection band. • The bandwidth defines the wavelength range over which the absorbance values are averaged to give the result at each time point. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 61 UV detectors which because of mechanical tolerances in moving gratings needed to avoid steeply sloping parts of the spectrum. Diode-array based detectors do not have this limitation Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 62 If there is no part of the spectrum with zero absorbance then it will be better to have the reference signal turned off. Figure 23 Spectrum of Anisic Acid Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 63 The light level will be lower so the signal will have more noise - the noise level depends on the wavelength and mobile phase solvents used. Figure 24 Benzene at 1 and 4 nm Slit Width (Principle) Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 64 If the resulting data rate is too slow (over filtering) the peak heights will be reduced and the resolution between them reduced; too fast and the data is noisier than it need be to accurately profile narrow peaks. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 65 0.0625 > 0.00625 0.125 > 0.0125 0.25 > 0.025 > 0.05 > 0.1 > 0.2 1.25 > 0.4 0.625 > 0.85 16.0 0.3125 Values in the User Interface may be rounded. Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 66: How To Achieve Lowest Carry Over

    This will be effective in removing a small degree of carryover but for more effective washing of the outside of the needle use the flushport. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 67 For samples where the outside of the needle cannot be cleaned sufficiently with water or alcohol from the flush pump use wash vials with an appropriate solvent. With an injector program several wash vials can be used for cleaning. Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 68: How To Prevent Column Blockages

    7 An in-line filter (1290 Infinity inline filter, diameter 2 mm, p/n 5067-4638) is recommended to catch particulates and extend column life. 8 Change the filter when the pressure increases by 10 %. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 69 14 If samples are dissolved in high organic content consider using an injection program to add a plug of weaker solvent before and after the sample aliquot to reduce the risk of precipitation in high concentration buffer mobile phase. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 70 Optimization of the Agilent 1290 Infinity LC System How to Prevent Column Blockages Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 71: System Setup And Installation

    Agilent 1290 Infinity LC System Manual and Quick Reference System Setup and Installation Installing Software Installing the Module Optimizing the Stack Configuration (Binary LC System) Optimizing the Stack Configuration (Quaternary LC System) Priming the Pump Purging the Pump Flow Connections Between Modules...

  • Page 72: Installing Software

    • Attempts to identify supported LAN-based instruments that are powered on and connected to your PC or lab’s network, • Automatically suggests replacements parts and troubleshooting tasks for some common instrument problems. Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 73: Installing The Module

    75). This configuration optimizes the flow path for minimum delay volume and minimizes the bench space required. The Agilent 1290 Infinity Binary Pump should always be installed at the bottom of the stack. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 74 System Setup and Installation Installing the Module Figure 25 Recommended stack configuration for 1290 Infinity with binary pump (front view) Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 75 System Setup and Installation Installing the Module Figure 26 Recommended stack configuration 1290 Infinity with binary pump (rear view) Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 76 A slightly longer capillary is required between the pump and autosampler. (See Figure 27 on page 76 and Figure 28 page 77). Figure 27 Recommended two stack configuration for 1290 Infinity with binary pump (front view) Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 77 System Setup and Installation Installing the Module Figure 28 Recommended two stack configuration for 1290 Infinity with binary pump (rear view) Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 78: Optimizing The Stack Configuration (Quaternary Lc System)

    80). This configuration optimizes the flow path for minimum delay volume and minimizes the bench space required. The Agilent 1290 Infinity Quaternary Pump should always be installed at the bottom of the stack. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 79 System Setup and Installation Installing the Module Instant Pilot Solvent cabinet Detector Column compartment Autosampler Pump Figure 29 Recommended stack configuration for 1290 Infinity with quaternary pump (front view) Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 80 LAN to control software CAN Bus cable to Instant Pilot Analog detector signal (optional) AC Power CAN Bus cable Figure 30 Recommended stack configuration for 1290 Infinity with quaternary pump (rear view) Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 81 Figure 32 page 82). Instant Pilot Detector Column compartment Solvent cabinet Pump Autosampler Thermostat for the ALS (optional) Figure 31 Recommended two stack configuration for 1290 Infinity with quaternary pump (front view) Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 82 CAN Bus cable to Instant Pilot Analog detector signal (optional) Thermo cable (optional) CAN Bus cable AC Power Figure 32 Recommended two stack configuration for 1290 Infinity with quaternary pump (rear view) Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 83: Priming The Pump

    Ensure that the pump is controlled by computer software or the Instant Pilot controller and that the flow rate is set to zero. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 84 The purge valve switches the flow path back to the system. It is recommended that each channel is now purged with 30 ml of propan-2-ol. Follow the procedure “Purging the Pump” on page 85. Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 85: Purging The Pump

    ➔ The volume of substances should be reduced to the minimum required for the analysis. ➔ Do not operate the instrument in an explosive atmosphere. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 86 1 To access the setup page for controlling the purge valve, right-click on the pump section, and select Control from the context menu. Alternatively, you can select Instrument > More 1290 Infinity BinPump > Control. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 87 Immediately following the purge, the solvent in the pump has not been degassed and so the system should be operated for at least 10 min so that the solvent is degassed. Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 88: Flow Connections Between Modules

    Max-Light Cartridge Cell in the 1290 Infinity detector and place the outlet in an appropriate waste collection container. Integration Into the Network For network integration of your system refer to user manuals of your modules (chapter LAN Configuration). Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 89: Quick Start Guide

    Agilent 1290 Infinity LC System Manual and Quick Reference Quick Start Guide About the Quick Start Guide Preparing the system Turning the System ON Loading the Default Method Configuring the Online Plot Purging the Pump Data Acquisition in Method and Run Control View...

  • Page 90: About The Quick Start Guide

    The starting point assumes that the system has been installed, turned on and initially N O T E primed (see “Priming the Pump” on page 83). The UV lamp should be turned on at least 30 minutes before starting any quantitative work. Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 91: Preparing The System

    System On/Off button in the bottom right of the system diagram. The system status changes from Not Ready (yellow indication) to Ready (green indication) after a short delay as the setpoints are attained. Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 92: Loading The Default Method

    Note that this method can never be overwritten with new parameters. Hence clicking on N O T E Save will re-direct you into the Save As... function, so that you must enter a different method name. Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 93: Configuring The Online Plot

    1 If the Online Plot window is not visible: Click View > Online Signals > Signal Window 1 to display the window. 2 To configure the desired signal(s) in the Online Plot window, click Change…. The Edit Signal Plot setup page opens. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 94 The Balance button zeroes all detectors when pressed. Changes made in the Online Plot page do not in any way affect the data stored into the N O T E individual data files. Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 95: Purging The Pump

    (isopropanol is often a good choice, check with a solvent miscibility table). For details on the purging procedure, refer to “Purging the Pump” on page 85. Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 96: Data Acquisition In Method And Run Control View

    Quaternary Pump. Method Parameters for Test Mixture and ZORBAX RRHD Column The 1290 Infinity LC System is supplied with a ZORBAX RRHD Eclipse Plus C18 1.8 µm, 2.1 mm x 50 mm, column and a test mixture containing phenones for use with this example procedure.
  • Page 97 1.8 µm, 2.1 mm x 50 mm i.d. Temperature 40 °C Detector Signal A 250 nm, bw 100 nm, ref 360 nm, bw 100 nm Peak width 0.025 min (10 Hz) Store Spectra Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 98: Setting Up The Method

    In the Method page for the 1290 Infinity Autosampler, enter the following parameters: • Injection volume: 1.0 µl • Injection with Needle Wash • Mode Flush Port, Time: 6 s Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 99 7 Allow the system to equilibrate for at least 10 min, and check that the baseline in the Online Plot is stable before starting the analysis. Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 100: Running The Method For A Single Injection

    The chromatogram should look similar to Figure 33 on page 101 — the time axis will be longer, however, as this was run under four times faster conditions in the next section. Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 101: Running The Method Faster

    The chromatogram should look similar to the one shown below, with a run time of about 1 min. Figure 33 Example chromatogram of the phenones test mixture using Quick Start conditions Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 102 • Examine the spectra of the peaks and select appropriate narrow band detection. For more details on optimization, please refer to “Optimization of the Agilent 1290 Infinity LC System” on page 39. Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 103: Data Analysis

    For more detailed information on data analysis including the use of calibration for quantification refer to the ChemStation manual. Method source Navigation table Integration task Spectrum task Navigation panel Figure 34 Data Analysis view Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 104: Data Analysis View

    Double click Single Runs to load these data files into the Navigation Table. 4 Select a file in the Navigation Table, and double click it to load the chromatogram into the viewer. Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 105: Integrating A Signal

    0.01 in this case. 6 Area Reject and Height Reject can be set to reject the smallest peaks. 7 Click the Integrate icon to update the results using these new settings. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 106 Quick Start Guide Data Analysis 8 Exit from the Events Table using the green tick icon (see figure below). Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 107: Specify The Report

    Quick Start Guide Data Analysis Specify the Report 1 On the menu bar click Report > Specify Report to display the window shown in the figure below. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 108 This completes a brief look at using the Data Analysis part of the ChemStation software. Consult the ChemStation manuals and the online help system for further details about the powerful features of the ChemStation. Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 109: Appendix

    Agilent 1290 Infinity LC System Manual and Quick Reference Appendix Safety Information Solvent Information Agilent Technologies on Internet Setting Up a Method using Edit Entire Method Method Information Instrument/Acquisition Data Analysis Run Time Checklist This chapter provides additional information on safety, legal and web and about setting up a method.

  • Page 110: Safety Information

    Any interruption of the protective earth grounding will cause a potential shock hazard that could result in serious personal injury. Whenever it is likely that the protection has been impaired, Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 111 When working with solvents, observe appropriate safety procedures (for example, goggles, safety gloves and protective clothing) as described in the material handling and safety data sheet by the solvent vendor, especially when toxic or hazardous solvents are used. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 112 ➔ Do not proceed beyond a caution until you have fully understood and met the indicated conditions. Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 113: Solvent Information

    THF, dioxane, di-isopropylether) such ethers should be filtered through dry aluminium oxide which adsorbs the peroxides, • Solvents containing strong complexing agents (e.g. EDTA), • Mixtures of carbon tetrachloride with 2-propanol or THF. Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 114: Agilent Technologies On Internet

    For the latest information on products and services visit our worldwide web site on the Internet at: http://www.agilent.com Select Products/Chemical Analysis It will provide also the latest firmware of the modules for download. Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 115: Setting Up A Method Using Edit Entire Method

    Edit Entire Method function as this automatically steps through the pages. This is accessed through the menu Method > Edit Entire Method, and opens the Check Method Sections to Edit dialog: Figure 35 Check Method Sections to Edit Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 116 If you inadvertently pressed OK before all entries have been made, then use Cancel and restart Edit Entire Method. Alternatively, carry on and return to the unfinished screen at the end. Clicking Cancel offers a button to Skip the remaining screens. Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 117: Method Information

    This box allows information about the method to be entered. This information will be displayed above the system diagram on the Method and Run Control screen whenever this method is loaded and resident in memory. Figure 36 Method Information Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 118: Instrument/Acquisition

    RC.Net drivers for the instrument modules. To run the example separation, as with most methods, it is not necessary to change every available parameter but for completeness they are described in the next sections. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 119 Flush Port or by dipping into a specified Wash Vial. • Time in seconds the peristaltic pump connected to the flush port will pump the rinse solvent. It then pumps for a further 15 s to clear the flush port. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 120 Automatic delay volume reduction (ADVR) switches the injection valve from mainpass to bypass after the injection has taken place and a volume defined by the sample flush out factor has swept through the injector. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 121 Injector Cleaning allows the injection system to be flushed with solvent. • Injection Valve Cleaning allows the valve to be switched at setpoints during the run to minimize carryover when problem compounds are injected. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 122 A commonly used example is the derivatization of amino acids with OPA and FMOC reagents. For details, please refer to the Agilent 1290 Infinity Autosampler manual. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 123 The pump will form binary mixtures of the selected A and B Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 124 Solvents section of the screen. • Show timetable graph displays the timetable changes graphically when the box is ticked. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 125 This is used as an alternative safeguard when the column is not inside a module with a leak sensor or in case the system is pumped dry. A value of 10 to 20 bar is typical. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 126 Selecting the As Detector Cell option will automatically read the temperature from the cell in the 1290 Infinity Detector. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 127 Three types of valve are available: • 2-position 6-port – used to switch between 2 columns • 2-position 10-port – used for alternating column regeneration • 8-position 9-port – used for multiple column selection in MDS Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 128 • Stop Time / Post Time are set to No Limit / Off and these values are usually taken care of in the Pump tab. However, the detector module Stop Time can sometimes differ from the pump Stop Time when it is required that data Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 129 For multiple signals it may be necessary to have signal A set as a broadband detector to ensure peak spectra are available for all the different wavelength signals. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 130 However, it should not be changed without good reason, as making it more negative will increase baseline noise and reduce the range available for measuring positive peaks. • Advanced - Slit Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 131 • Advanced - Lamps on required for analysis: the 1290 Infinity DAD or MWD has one deuterium lamp and it needs to be on for analysis so this box should be checked. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 132 A/B Composition — can be useful for overlaying the gradient profile on a chromatogram. • Thermostatted Column Compartment: • Left/Right temperature • Detector: • Board temperature • Optical unit temperature • UV lamp anode voltage Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 133: Data Analysis

    If a system uses multiple detectors such as the diode-array detector and a mass spectrometer, the Signal Description lines allow delay times for the downstream detector to be entered so that the software can align the peaks from the different detectors. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 134 Appendix Setting Up a Method using Edit Entire Method Figure 43 Signal details Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 135 Data Analysis view. The integration events can be optimized at that time and default settings are often used for the initial acquisition runs. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 136 Report icon and then selecting Specify Report in the context menu. In the Data Analysis view it can be accessed through the menu Report > Specify Report or the Specify Report task icon. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 137 To setup a simple area% report with Classic Reporting, which prints to the printer and to a PDF file, enter the following settings in these sections of the Specify Report screen: Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 138 On the Quantitation settings tab: • Calculation mode • Calculate: Percent • Based on: Area Click OK to exit and the next screen in the Edit Entire Method process will open. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 139 Setting Up a Method using Edit Entire Method Instrument Curves Figure 46 Instrument Curves screen The Instrument Curves checkboxes allow these recorded parameters to be overlaid as a graph on the chromatogram. Agilent 1290 Infinity LC System Manual and Quick Reference...

  • Page 140: Run Time Checklist

    The software looks for the macro in the C:\Chem32\Core directory; include the path if it is located elsewhere. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 141 Checklist and from the Edit Entire Method process. The method should now be saved into the master methods directory, by default C:\Chem32\1\Methods, using File > Save As > Method or Method Menu > Save Method As. Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 142 Sub-2-micron particles single injection configuration high throughput one stack Optimization two stack front two stack rear network integration two stack Infinity Autosampler configuring Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 143 Thermostatted Column Compartment description report specification van Deemter Equation resolution Optimization response time wavelength and bandwidth retention factor optimization Run Time Checklist safety class I Agilent 1290 Infinity LC System Manual and Quick Reference...
  • Page 144 Agilent 1290 Infinity LC System. The manual describes the following: • introduction, • product description, • system optimization, • setup and installation, • quick start guide.  Agilent Technologies 2009-2011, 2012 Printed in Germany 05/2012 *G4220-90301* *G4220-90301* G4220-90301 Agilent Technologies...

Table of Contents