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  • 1.  Decrease in MS response of SIL internal standard with increasing analyte concentration

    Posted 10-21-2025 15:28

    Hi all,

    We are using a SIL internal standard for LC-MS assay development to quantify a small molecule analyte. In our calibration curves, there's a systematic DECREASE in the MS signal of SIL IS as the analyte concentration increases in calibrators. It appears as if the analyte is interfering with the ionization of the SIL IS.

    Have you ever seen a phenomenon like this? Any recommendations on how to solve it?



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    Babak Basiri
    Director, ADME/PK
    Dyne Therapeutics
    Waltham MA
    [email protected]

    Disclaimer: Opinions expressed are solely my own and do not express the views or opinions of my employer.
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  • 2.  RE: Decrease in MS response of SIL internal standard with increasing analyte concentration

    Posted 10-22-2025 09:11

    Hi Barak, 

    We run into this situation on occasion and in most cases, we are able to reduce the SIL IS concentration in the final extractions in order to mitigate the impact.  Sometimes even a 2-fold decrease in the final IS concentration is enough to eliminate or even just minimize the impact.  Hopefully this relatively quick solution will work for your assay as well. 

    Thanks!

    Katy



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    Kathleen McQuate
    Research Scientist
    Charles River Laboratories Ashland LLC
    Ashland OH
    [email protected]

    Disclaimer: Opinions expressed are solely my own and do not express the views or opinions of my employer.
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  • 3.  RE: Decrease in MS response of SIL internal standard with increasing analyte concentration

    Community Leadership
    Posted 10-22-2025 09:30

    Hi Babak,

    We've encountered this issue before - ISTD suppression at high analyte concentrations due to ionization competition.

    Perhaps try: 

    (1) Alternative ionization techniques (e.g., switching between ESI+/- or APCI)
    (2) Mobile phase modifiers - consider using ammonium salts
    (3) Dilution, followed by injection on a more sensitive MS

    Good luck!

    Stephanie 



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    Stephanie Cape Ph.D.
    Senior Director, Precision Medicine Delivery
    Labcorp

    [email protected]

    Disclaimer: Opinions expressed are solely my own and do not express the views or opinions of my employer.
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  • 4.  RE: Decrease in MS response of SIL internal standard with increasing analyte concentration

    Posted 10-23-2025 00:57

    Thank you, Katy and Stephanie for your suggestions. Hopefully one of these options works for us!

    Best,

    Babak 



    ------------------------------
    Babak Basiri
    Director, ADME/PK
    Dyne Therapeutics
    Waltham MA
    [email protected]

    Disclaimer: Opinions expressed are solely my own and do not express the views or opinions of my employer.
    ------------------------------