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new column

Mr.Good

This is my first time dealing with Agilent's AdvanceBio AAA amino acid chromatography column. Due to my carelessness, I left the mobile phase A to flow through the newly purchased column at a low flow rate overnight. The next morning, the column already showed high pressure blockage. May I ask how should I salvage my new column? Is a 50% acetonitrile water solution effective?

  • +1

    Hi Mr good,

    I suppose you mean with phase A the same as in this compendium:

    https://www.agilent.com/cs/library/applications/compendium-%20aminoacid-advancebio-5994-0033EN-us-agilent.pdf

    Then following advise will be the one to follow:

    Was the column running continuously during these experiments (non-stop injections) or were there periods when the column was left ?  It is particularly important not to leave the column in Mobile Phase A for any length of time.  The column should be left in Mobile Phase B or, better still, flushed into 90% ACN for storage.
    Any contamination of the column is best addressed by (1) regularly replacing the guard column; (2) regular washing of the with stronger organic eluent (e.g. 90% ACN is stronger than 45% ACN + 45% MeOH); (3) flushing the column in the reverse direction to unclog frits ... this should not be necessary if the sample is properly dissolved and has been filtered before injection
  • 0 in reply to ivanderm

    I am very pleased that you are able to provide me with a very good answer. Yes, the information you provided is very accurate and helpful. I tried to analyze why there was excessive pressure (in fact, my samples were strictly processed), and I attempted to flush the chromatography column with a mobile phase of 45% ACN + 45% MeOH, but the results were not satisfactory. So after approximately 40 column volumes, I stopped the operation. Next, I will follow your advice and try using 90% ACN. I will continue to share any progress here. Thank you again for your attention and answers.

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