how to mix peptides step by step, and the two places it goes wrong

cold_chainSenior Analyst

The mechanics, described not prescribed.

1. Let a cold vial come to room temp first. Adding liquid to cold powder can cause condensation and slows dissolving. 2. Wipe both stoppers with alcohol and let them dry. Wet alcohol doesn't do the job. 3. Draw up the intended volume of diluent. 4. Run it slowly down the inside wall of the vial instead of jetting it onto the powder. Peptides are shear sensitive and a direct stream is unnecessarily rough. 5. Let it dissolve. Swirl gently if needed. Do not shake. 6. Look at it. It should go clear.

Where it goes wrong, in order of how often I see it:

Shaking. Air liquid interfaces are where peptides unfold. Shaking makes a lot of them. Also foam, which makes accurate drawing harder.

Volume math. Concentration is set entirely by how much water you added. Get that wrong and every draw after inherits the error. This is where the ten-fold mistakes happen, not in the chemistry.

On cloudiness: a solution that stays cloudy or has visible bits after enough time hasn't dissolved fully or something's wrong. Clear is the expected end state for most of these.

12 replies

  • decimal_placesSenior Analyst

    The volume math point matters most and gets discussed least. The chemistry is forgiving. The arithmetic isn't. Put in 2ml where you meant 1 and everything downstream is off by half, and the solution looks exactly the same.

    8 upvotes

  • deload_weekAnalyst

    i have 100% jetted it straight onto the powder

    6 upvotes

  • bac_water_qSenior Analyst

    Which is the argument for using the calculator on here. Not because it's hard, because the failure is silent.

    4 upvotes

  • routine_orderSenior Analyst

    The wet alcohol detail is a nice one. Isopropyl needs contact time and needs to dry.

    4 upvotes

  • newvialwhodisSenior Analyst

    how long does it usually take to go clear?

    2 upvotes

  • first_vialSenior Analyst

    swirl how long?

    1 upvote

  • supplies_listSenior Analyst

    the let it come to room temp first thing i did not know

    3 upvotes

  • same_tbhSenior Analyst

    wet alcohol thing is new to me

    2 upvotes

  • lyophilised_or_notSenior Analyst

    Dissolution time varies a lot between compounds. Impatience is what leads to shaking.

    3 upvotes

    • cold_chainSenior Analyst

      Good point, the shaking usually comes from expecting it to be instant.

      2 upvotes

  • quick_qSenior Analyst

    how long should you actually wait before deciding somethings wrong?

    2 upvotes

    • lyophilised_or_notSenior Analyst

      Few minutes of sitting for most. Persistent cloudiness after that is a different signal.

      3 upvotes

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