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
      Molecular Spectroscopy
      • Molecular Spectroscopy - UV-Vis & UV-Vis-NIR, FTIR, Raman & Fluorescence
      • 8700 LDIR User Group
    • 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
Consumables
  • Technical Areas
Consumables
Wiki Best Practices for Aqueous Mobile Phases
  • Announcements
  • Forum
  • Files
  • Wiki
  • More
  • Cancel
Actions
  • Share
  • More
  • Cancel
Table of Contents
  • Consumables
  • A Beginner’s Guide to Hydrophobic Interaction Chromatography
  • A simple tip to protect your columns
  • A Tip for Preparing Robust and Consistent Mobile Phases
  • Achieve accurate and repeatable gas flow meter measurements
  • AdvanceBio Columns Blog Series
  • Agilent 123 Meter Sodium probe maintenance
  • Agilent Collection of Columns, Supplies, and Standards Resources
  • Analysis of microplastics in the environment
  • Analyzing ADCs by HIC
  • Avoiding downtime in the lab: top tips for GC/MS success
  • Best Practices for Aqueous Mobile Phases
  • Best Practices for Making Good Connections
  • Bio LC Column User Guides
  • Bursting Tubing and Columns (GC and HPLC)
  • Calculate the Maximum Allowable Working Pressure for Tubing
  • Calculating Column Volume
  • Cannabis Potency Testing: a Reliable, Cost-Effective Method
  • Carbohydrate Analyses in LC
  • Checking Your Intuition - Sub 2 µm vs Superficially Porous
  • Choosing the right GC Injection Technique
  • Choosing the right pore size for size exclusion chromatography
  • Columns, Supplies, and Standards Knowledgebase
  • Consumables Applications and Workflows
  • Consumables Recommended Supplies Lists for Agilent.com
  • Custom Product Request
  • Extreme Makeover – Derivatizations in Chromatography – Part 1 GC
  • Extreme Makeover – Derivatizations in Chromatography – Part 2 LC
  • Fake It Until You Make It: When BioInert Isn’t an Option
  • Flipping Amino Acid Analysis on Its Head
  • Glycans at a glance:  Analyzing therapeutic glycoproteins
  • Handle and Care of Syringes
  • Help! My Peaks Look Strange - Fronting and Tailing in GC
  • Help! My Peaks Look Strange - Saddle Points - LC/GC Troubleshooting
  • How do I select a Split/Splitless liner?
  • Hydrophobic Interaction Chromatography of Proteins and mAbs
  • Importance of Silica Particle Strength for Sub-2 µm SEC Columns
  • KB: Ferrules recommended for GC self-tightening column nut
  • LC and LC/MS Columns - USP Designations
  • LC Column User Guides
  • LC Method Translation - the Dwell Volume
  • Minimize spectroscopy workflow disruptions
  • Minimizing Metals for Best HILIC Results
  • More than just a drink: Analyzing the elemental composition of beer
  • Multi-Attribute Methods – Peptide Mapping Part IV
  • Must See Webinars
  • Nomenclature of CFC's/Freons/Halons/Coolants
  • Oligonucleotide Analysis - Unexpected Details Matter
  • Optimizing Bonding Chemistry for Sub-2 µm SEC Particles
  • Pass the Salt, Please – Mobile Phase Preparation for HIC
  • Pesticides and their stability during GC analyses
  • Pre-Columns - the forgotten art of using retention gaps
  • Problematic polar analytes? Hello HILIC…
  • Protecting your laboratory productivity
  • Recommended Reading
  • Sample Prep Pointers - Peptide Mapping Part I
  • Save your results with sample filtration
  • Simplified cone inspection with the new Agilent LED measuring magnifier
  • Software - Supported Method Development - The Scanview Application
  • Software tool for the ADM Flow Meter (G6691A)
  • Stay Safe: A Win-Win for Solvent Storage
  • Streamline your sample processing
  • The importance of chemical composition for vial performance
  • Tips & Tricks for Amino Acid Analysis – Part I
  • Tips & Tricks for Amino Acid Analysis – Part II
  • Tips & Tricks for Amino Acid Analysis – Part III
  • Tips & Tricks for Amino Acid Analysis – Part IV
  • Tips for Smooth Sailing with HIC
  • Troubleshooting HPLC autosamplers
  • Troubleshooting HPLC degassers
  • Troubleshooting Sequence Coverage – Peptide Mapping Part III
  • UltiMetal Plus Flexible Metal Ferrule
  • UV, MS, TFA, and Formic Acid – What to use? Peptide Mapping Part II
  • What are the typical % Gain or EHT values for hollow cathode lamps?
  • You Need Lamps or Chemical Standards for Atomic Absorption Single-Element Analyses?
Still Need Help?

Post your question in our User Forum or Contact Support.

Best Practices for Aqueous Mobile Phases

Created by anne_blackwell anne_blackwell over 2 years ago | Last modified by Agilent Agilent over 2 years ago

 

Aqueous mobile phases are ripe for microbial growth.  This post discusses ways to prevent microbial growth from interfering with your chromatography.

 

Picking the right mobile phases can be key to developing a robust method – but good mobile phase hygiene is just as important! 

 

Aqueous mobile phases are great breeding grounds for microbes.  These microbes can lead to clogging at any point of the flow path, from pump chambers to capillaries to LC columns.  Some of those points of clogging are easier to replace or clean out than others. 

 

Here’s what aqueous mobile phase looks like when it’s been left to sit for 6 months:

(Photos courtesy of my colleague Bill)

 

You may say you’d be crazy to use that mobile phase, but I have colleagues who have seen people do it!  Joking aside, microbial growth will cause problems long before it becomes visible to the naked eye, so we must take steps to prevent it even for mobile phases we only plan to use for a couple days.

 

Effective ways to mitigate microbial growth are to include organic solvents or additives like azide to the solution.  For a reverse-phase method where the initial conditions already include some portion of organic solvent, making Mobile Phase A 95/5 H2O/ACN may be a reasonable approach, but for 100% aqueous chromatographies like size exclusion or ion exchange, adding organic solvent simply isn’t an option.  Azide is highly toxic, making it an unappealing solution, and some countries and laboratories ban its use outright.

 

Aqueous mobile phases should be filtered through a 0.45 or 0.2 µm filter to remove microbes and other particulates, after all buffer salts have been dissolved and the pH has been adjusted.  (Really, get out that stir bar and stirrer/hotplate if necessary.)  Refrigeration can help extend lifetime for up to a few weeks, but at room temperature aqueous mobile phase should be remade fresh every other day at a minimum.  As an extra measure, you could add an in-line filter to your LC system, either between the pump and autosampler or immediately before the column depending whether or not you want to filter the sample as well.

Until next time!

 

Anne

 

 

Keywords: Biocolumns, LC, Mobile Phases, Best Practices

  • Best practices
  • lc mobile phase
  • bio columns
  • Biopharma LC Columns & Consumables
  • liquid chromatography
  • Share
  • History
  • More
  • Cancel
  • Sign in to reply
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
Related
Privacy Statement
Terms of Use
Contact Us
Site Help