Fast(er) SEC- is there a way to stagger injections and data collection?

ShellyBuhay
ShellyBuhay
in Instrumentation and Mass Spectrometry
<p>We have been looking at ways to speed up our size exclusion assays, for faster throughput for screening of formulation samples.  We have looked at the Acquity BEH SEC column, and found it to work great - immediately cut our run times by at least half, and possibly more as we work to optimize the method. </p><p></p><p>Looking at what else we can do, I recently came across an article that made a valid point- for an SEC column, all of the separation comes between the exclusion limits and the inclusion limits of the column, so it is really unnecessary to collect data until you reach the exclusion limit.  Their idea was to stagger injections; for example, for a conventional column, you would inject, wait ~10 minutes (depending on the properties of your column), and then while you are acquiring data for the first sample where the sample actually separates,  inject the sample again, so that you are constantly collecting data over the ~15 minutes or so where the separation is actually happening (I hope that makes sense).  This would significantly cut run times, and would be really great for rapid screening.</p><p></p><p>Thinking about the options on the UPLC, though, I'm not sure if it would be possible to run the instrument this way.  I'm hoping a more expert user might weigh in as to how it may be done.</p><p></p><p>The paper I was looking at is: J Chromatog A 1216 (2009) 8904-8909.</p><p></p><p>Thanks!</p>

Answers

  • MarcNoble
    MarcNoble

    Marc Noble

    Principal Service Specialist

    Phone - 800-252-4752 x6220

    Mobile - 484-363-7814

    Email - marc_noble@waters.com

  • lizh
    lizh

    Interesting...wait til the rallies to find out...no seriously,we are looking at Multipump config for PittCon, but here's the rub. If only it had been

    any other column. SEC could be used with a MALLS detector and I needed to check the backpressure recovery and whether it i feasible. My concern is

    that there might be an impact on the column. Anyway I have sent out a call to Beth Gildea and the bio team and I will let you know.

    Liz

    New message: "Fast(er) SEC- is there a way to stagger injections and data collection?"

    Marc Noble

    to:

    Elizabeth Hodgdon

    01/12/2011 01:39 PM

    Please respond to p-reply

    Fast(er) SEC- is there a way to stagger injections and data collection?

    reply from Marc Noble in ACQUITY UPLC - View the full discussion

    Marc Noble

    Principal Service Specialist

    Phone - 800-252-4752 x6220

    Mobile - 484-363-7814

    Email - marc_noble@waters.com

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  • PeteClaise
    PeteClaise

    Hi Shelly,

         Thanks for your question.Unfortunately at this time it is not possible to perform this procedure as described in the paper. Empower will not start a new injection, i.e., draw sample, until the previous injection run time is complete.

    Best Regards,
    Pete

  • ShellyBuhay
    ShellyBuhay

    Thanks Pete - that's what I thought, but I wanted to be sure!

    Shelly.

  • Southpaw
    Southpaw

    That's actually untrue, you can certainly stack injections on Waters systems.  We've successfully done it on the Alliance systems and it should lend just fine to Acquity systems too.  All you have to do is set up two instruments, one that contains the separation module (pumps, column heater, etc) and one that contains the detector module(s).  You can then use the contact relays on the separation module to initiate the detection window and tune your runtime in both the separation and detection "instruments" to have stacked injections.

    Feel free to PM me for specific information.

  • ShellyBuhay
    ShellyBuhay

    Oh, that's clever!  I can't wait to give that a try (but it will be a few weeks before I can get some time on the instrument).  Thanks for the great idea!

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