Agilent Community
Agilent Community
  • User
  • Site
  • Search the Community
  • User
  • Home
  • Technical Areas
    Technical Areas
    • AssayMap
    • Atomic Spectroscopy
    • Automated Electrophoresis
    • Bioreagents
    • Cell Analysis
    • Consumables
    • CrossLab Connect
      CrossLab Connect
      • Service Management
      • Smart Alerts
    • Dissolution
    • GC
    • GC/MS
    • LC/MS
    • LC
    • Molecular Spectroscopy
    • Sample Preparation
    • Software
      Software
      • Chromatography Software
        OpenLab CDS, OpenLab ChemStation, OpenLab EZChrom
      • Data Management Software
        OpenLab ECM, OpenLab ECM XT, OpenLab Server
      • Mass Spectrometry Software
        Data Analysis, Data Acquisition, Productivity App, WalkUp Software
    • Vacuum
  • Applications Areas
    Applications Areas
    • Basic Research
    • Energy and Chemicals
    • Environmental
    • Food and Agriculture
  • Agilent Knowledge Portal
  • Links
    Links
    • Collection of Support Resources
    • Webinar Notifications
    • Contact Support
    • User Contributions
  • Getting Started
    Community Help and Resources
  • Browse
    Browse
    • People
    • Places
    • Your View
    • Your Community Inbox
LC Portal
  • Agilent Knowledge Portal
LC Portal
Articles Troubleshooting Peak Area Variation Between Injections
  • Articles
  • Videos
  • Forum
  • Announcements
  • More
  • Cancel
  • Agilent Knowledge Portal
  • LC Portal Home
  • LC Portal Articles
  • LC Portal Videos
  • More
  • Cancel
  • Created :  10 Nov 2022
  • Modified :  6 Feb 2023
  • Category :  Agilent Knowledge Portal
  • Entry Type :  Article
  • Product Type :  Analytical LC Systems
  • Component :  LC Injectors and Sample Thermostats
  • Product Name :  1100 Series LC System 1120 Compact LC 1200 Series LC System 1220 infinity II LC System 1260 Infinity II Bio-Inert LC System 1260 Infinity II High-Temperature GPC System 1260 Infinity II LC System 1260 Infinity II Multi-Detector GPC/SEC System 1260 Infinity II Prime Bio LC System 1260 Infinity II SFC System 1260 Infinity II SFC/UHPLC Hybrid System 1260 Infinity LC System 1260 Infinity II Prime LC System 1290 Infinity II Bio LC System 1290 Infinity II GPC/SEC System 1290 Infinity II LC System 1290 Infinity LC System
  • Task :  Troubleshooting
Related Articles
  • BAC Method Missing from the Agilent Femto Pulse Software

    This Information Applies To: Agilent Femto Pulse, Agilent Femto Pulse controller software 1.0, ProSize analysis software 3.0, Agilent Genomic 55 kb BAC kit (p/n FP-1003-0275 ) Issue: In Agilent Femto Pulse controller software version 1.0, it is not possible to select the FP-1003-22 55 kb BAC method from the Separation Set up dialog box. This situation occurs due to the BAC method and configuration files not being included in the Femto Pulse controller version 1.0 and ProSize 3.0 software installers...
  • How to Export Agilent TapeStation Logs

    This Information Applies To: 4200, 4150 and 2200 TapeStation instruments, TapeStation analysis software, TapeStation controller software Issue: Technical support require the instrument log files to troubleshoot hardware issues with the Agilent 4200, 4150 and 2200 TapeStation systems. It is possible to export the logs via the TapeStation controller or analysis software. Steps to follow Tip : Log files do not include data files, experimental, or sample information. Collect and...
  • How to replace the peristatic pump on 1290 Infinity (G4226x) and High Performance Autosamplers (G1367x)

    This Information Applies To: 1290 Infinity (G4226x) and High Performance Autosamplers (G1367x) Issue: The Peristaltic Pump needs to be replaced if it has stopped working due to worn tubing, if it is leaking, or as part of annual maintenance of the autosampler. Steps to follow: Warning : Make sure that appropriate personal protective equipment to manipulate the chemicals used in this process. Nitrile gloves (or equivalent) and safety glasses are recommended. ...
  • Determining which Module is Causing the Shutdown Error Due to a Leak

    This Information Applies To: Agilent LC Systems Issue Agilent modules will go into an error status (red status light) when an error condition specific to that module occurs. For example, metering home failed will be specific for the autosampler, a lamp ignition failure will be specific for a detector, or a pump stroke will be specific for the pump. In those cases, only the module with the error will go into an error status. However, there are some conditions that will put all the modules into...
  • LED Status Indicators

    This Information Applies To: All Agilent LC modules Issue: The LED Status Indicators are common features for all the HPLC modules. The indicators are located on the top right corner of each module, opposite to the power switch led as shown in Figure 1 . They can vary between six different statuses: Grey (led off), Green, Yellow, Red, flashing Yellow or flashing Red. The LED Status is very informative on the module performance and can provide important information for troubleshooting. ...

Troubleshooting Peak Area Variation Between Injections

Answer

This Information Applies To: All Agilent HPLC Autosampler modules


Issue:

Nonreproducible peak areas between injections, using the same sample and the same experimental conditions.


Resolution: 

The issue may come from different sources, it is recommended to verify all the following ones for a successful troubleshooting.

 Warning 
Warning: When opening capillary or tube fittings, solvents may leak out. Observe appropriate safety procedures (for example, wear goggles, safety gloves and protective clothing) as described in the material handling and safety data sheet supplied by the solvent vendor, especially when toxic, or hazardous solvents are used.
  • Vials
    Use only Agilent vials, the shape, and the material is optimized for maximizing instrument precision.
    Do not overfill the vials. Fill the vial up to 1/2 to 2/3 of the total volume.

  • Experimental conditions
    Make sure that the same solvents were used between the runs.
    Keep the room temperature constant and under 25 degrees Celsius.

  • Acquisition method settings
    Optimize the draw speed decreasing it down to 50µL/min. The default settings are at 200µL/min, but this speed may be too high for viscous samples.

  • Hardware
    The issue may reside in the Autosampler. A leakage is one of the frequent causes, make sure that all the autosampler connections are tight and there is no leaks between the needle and needle seat.

If the problem persists, contact Agilent Technologies for further guidance.

 Tip 
Learn how to effectively troubleshoot your LC autosampler:
HPLC-0GEN-2201e - Chromatographic Troubleshooting for HPLC e-learning course available from Agilent Education

 

 
 
Was this helpful?
  • Yes
  • No
Thank you for your feedback!

Agilent Community Feedback

Agilent Community Feedback

×
We are glad this was helpful! We are sorry this was not helpful. If you still need assistance please create a community post or contact support. To help us improve, please provide any additional feedback. For full details of how we will treat your information, please view our privacy policy.
Submit Cancel
Submit Cancel
Contact Agilent Support

Still Need Assistance?

Contact Agilent Support >

Privacy Statement
Terms of Use
Contact Us
Site Help