I have an Ultra Quat column and I know how to use it….

Are those words you wish you could say? Most recently, we seem to encounter more customers asking for advice in preparing a mobile phase for their Diquat and Paraquat analysis (related to EPA Method 549). There are a couple of different scenarios for this analysis. Please note that the procedures described here are intended for use with a modified version of EPA Method 549, and the mobile phases are not the same.


Case #1- Using HPLC with UV detection.

Perhaps this is a new project for you or you might have used our Ultra Quat Mobile Phase Modifier in the past and you want to use something that works similarly in conjunction with your Ultra Quat HPLC column (150 x 4.6 mm, 5 um, catalog# 9181565 in this case). Below is the suggested procedure to make mobile phase, taken from one of our Technical Bulletins.


Ultra Quat Reagent Solution preparation instructions:


Obtain sodium hexafluorophosphate (NaPF6), purity 98% or greater, such as currently offered by Sigma Aldrich and other vendors.

* Caution statement: neat material is corrosive and hygroscopic – read the MSDS for proper handling and storage.


For 1 liter of Ultra Quat Reagent Solution, used to aid the separation of paraquat and diquat on your Ultra Quat HPLC Column, follow these instructions:

  1. Using an analytical balance weigh out 5040mg of Sodium Hexafluorophosphate into a clean and dry glass vial.
  2. Rinse 1 liter volumetric flask with HPLC Grade water to remove any contamination.
  3. Transfer the Sodium Hexafluorophosphate from vial to the 1 liter volumetric flask.
  4. Triple rinse vial with HPLC Grade water and transfer rinsate to flask.
  5. Bring flask to volume with HPLC Grade water, stopper, and shake well to dissolve – solution is now ready for use.
  6. Solution has limited stability. Use prepared solution as soon as possible.Use this prepared solution as “Mobile Phase A”. Use 100% acetonitrile as “Mobile Phase B”.  Here are other chromatographic conditions:
  • Isocratic Flow at 95% A: 5%B, flow rate 1.0 mL/min
  • UV @ 257nm Paraquat
    UV @ 308nm Diquat
  • Injection Volume 20 uL


Case #2- Using LCMSMS

Since this is a mass spec method, you will need to use a mobile phase modifier that is compatible. Our best suggestion is to follow the procedure as indicated in the following application note. The mobile phase modifier in this case is heptafluorobutyric acid, prepared at 10 mM, combined in a 95:5 ratio with acetonitrile.  LCMSMS for this method is usually done with a column of smaller dimensions, catalog # 9181352 (50 mm x 2.1 mm ID, 3 µm).


For information on analysis of diquat and paraquat using our Raptor HILIC-Si column, please see the newer post “Have there been any new applications developed for Diquat and Paraquat?“.


We hope that this blog clears up some questions you might have.  If you have any further questions, contact us in Technical Services.


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4 Responses to “I have an Ultra Quat column and I know how to use it….”

  1. Amin H says:

    Does the heptafluorobutyric acid phase needs to be in Amber bottle?

  2. Hello Amin. The neat material usually packaged in an amber container to maximize the lifetime, since HFBA is somewhat light sensitive. Although it would be less critical for the diluted material in mobile phase, using an amber bottle would be a good protective measure if you have such a container available.

  3. Paul says:

    What is the function of the hexafluorophosphate in the mobile phase? And are there any alternatives that can be used in it’s place if NaPF6 is not available?

  4. Hello Paul. NAPF6 is a chaotropic agent. You can read more about that here: https://en.wikipedia.org/wiki/Chaotropic_agent
    We have found no other additive that works as well for this phase with this application. However, we do have a new column phase and method developed with the Raptor HILIC-Si column. There will be another blog post coming in the near future regarding that.
    Thank you for asking and hope all goes well for you.

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