CWU Chemistry Laboratory Specific Standard Operating Procedure
SOP for: (check one)
Process
Hazardous Chemical
Hazard Class of Chemicals
Name of Procedure: Working with a Sure-Seal bottle: Handling air- and moisture-sensitive reagents
Applies to Lab (name): Chamberland Research Group
SCI Room #: 313
Prepared By: James Seigenthaler, Stephen Chamberland
Date: 9/6/12
Revision: 9/11/12
Special Notifications:
Sure-Seal bottles are designed to protect air- and moisture-sensitive reagents from the water and oxygen present in the
atmosphere. Users must take proper precautions to protect themselves and the reagents from harm.
Hazard Description:
Air- and moisture-sensitive reagents are often reactive and harmful if exposed to the atmosphere, requiring the use of an
inert-atmosphere double manifold and proper anhydrous transfer of liquids using a syringe. Some air- and moisturesensitive materials are pyrophoric, meaning that they ignite spontaneously on contact with air. These chemicals must
be handled with extreme caution. Any deviation from the proper technique when using these chemicals can cause
serious injury or death.
Protective Equipment:
- Lab coat
- Protective wrap-around goggles
- Chemically resistant nitrile gloves (wear two pair of regular gloves or heavy-duty gloves when working with toxic,
highly toxic, or pyrophoric chemicals)
- Long hair tied back
- Close-toed shoes
- Full-face shield (if working with pyrophoric chemicals)
- Buttoned up flame-retardant lab coat instead of a regular lab coat (if working with pyrophoric chemicals)
Engineering Controls:
Work with air- and/or moisture-sensitive reagents only in a working chemical fume hood. Only expose the reagents to
inert atmosphere (e.g., N2 or argon gas) using a double manifold.
Special Handling or Storage Requirements:
Air- and moisture-sensitive reagents and solvents are stored in accordance with the hazard they present. The most
hazardous substances are usually stored in the freezer, but sensitive solvents are usually stored on the bench or in a
flammable cabinet. Keep these reagents away from water, and make sure they are stored and sealed properly after
use.
Spill Response Procedure:
Consult the SDS of the chemical being used for more information.
Decontamination/Waste Disposal:
Consult the SDS of the chemical being used for more information. See the SOP “cleaning syringes and syringe
needles” for instructions on cleaning syringes and needles used when handling air- and moisture-sensitive materials.
Special Emergency Procedures:
Consult the SDS of the chemical being used for more information.
Procedure may be performed after normal working hours (M-F, 8-5)?
Procedure may be left unattended?
PRINTED 2/9/2016
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CWU Chemistry Laboratory Specific Standard Operating Procedure
Step by Step Procedure: Also see the accompanying document “Handling Air-sensitive reagents – Aldrich Technical Bulletin AL-134”
and the three videos “How to Handle Pyrophoric Reagents” produced by the University of California, San Diego
(UCSD) before using a Sure-Seal bottle for the first time. You can find these videos on YouTube.
*Though all the precautions discussed in the videos are necessary when handling pyrophoric reagents, not all
procedures will be needed when using non-pyrophoric materials.
1. Turn up the nitrogen flow slightly (about 2-5 bubbles/second), but not so high to make the oil splash violently in the
bubbler.
2. Secure the bottle if necessary clamping it to a ring stand or to the metal bars in the fume hood using a sturdy metal
three-prong clamp.
3. Unscrew the red bottle cap, remove the Teflon tape or wax protecting the opening in the bottle cap, and then insert a
needle from one of the tygon tubes attached to the double manifold into the bottle, but not far enough to touch the
liquid level. This will introduce nitrogen to the bottle. If using a chemical that is stored in the freezer, allow it to
warm up to room temperature under nitrogen by carrying out this step after removing the bottle from the freezer.
4. Purge the syringe with inert gas to remove all the air from the syringe and needle. To do this, first insert the
appropriate syringe needle through the soft part of the Sure-Seal cap using one of the existing holes, if possible.
See the SOP “using a syringe to dispense liquids” if you have questions.
5. Slowly pull out the plunger, drawing nitrogen gas into the syringe. Watch the bubbler and ensure that oil is not drawn
back up too quickly due to the negative pressure of withdrawing the plunger. If necessary, withdraw the plunger
more slowly or increase the flow of the nitrogen gas.
6. Once the syringe is full of nitrogen gas, remove the needle from the bottle, and depress the plunger to eject the
nitrogen gas into the fume hood. Repeat this purging sequence two more times.
7. After purging the syringe with nitrogen gas, insert the needle into the bottle so that it is below the level of the liquid.
NEVER INVERT A BOTTLE OF CHEMICAL WHEN USING A SYRINGE. Holding the syringe so that the plunger
points toward the floor of the hood (see picture), then gently and slowly pull the plunger drawing the liquid into the
syringe. Go slow, and do not let any air bubbles come in through the plunger if using a glass syringe. If a gas pocket
is observed, remove it by pressing the plunger to force the bubble out of the syringe. Resume drawing the plunger to
bring in more liquid inside the syringe, repeat pressing the plunger if necessary to remove any gas. The syringe
measures by volume delivered, so if 5 mL is needed, fill the syringe with MORE than 5 mL of liquid.
8. Once slightly more than the desired volume is in the syringe, depress the plunger until the line on the plunger is
even with the gradations indicating the precise volume of liquid needed
9. Gently pull the needle out slightly so that the tip is above the level of the liquid in the bottle. Withdraw the plunger
until a small amount of gas is seen in the syringe. Now the chemical in the syringe is under a nitrogen atmosphere
and the tip of the syringe is protected by a blanket of nitrogen gas rather than being exposed to the atmosphere.
10. Gently remove the needle from the bottle, and insert it through the septum of the receiving or reaction flask. The
plunger should always be pointing down toward the floor of the hood. (See picture)
11. Now press the plunger slowly to dispense the liquid into the receiving flask. DO NOT pull the plunger back out and
squirt in any remaining liquid located in the needle; that volume is extra from the amount measured.
12. Gently remove the syringe, and rinse it out well in a nonreactive solvent such as CH2Cl2. Repeatedly withdraw and
depress the plunger to fill and empty the syringe several times. Once rinsed, the syringe is ready to be fully cleaned.
See the SOP “cleaning syringes and syringe needles.” This step does not need to be done if a non-reactive material,
such as a solvent, is being used. If a small amount of unreactive solvent remains in the syringe after the liquid has
been dispensed, disassemble the syringe in the fume hood and let the residual liquid evaporate.
13. After rinsing out the syringe, remove the nitrogen needle from the Sure-Seal bottle, place two strips of Teflon tape
across the opening in a cross (+) pattern (or use melted wax to seal the bottle), replace the cap, then store the bottle
where appropriate.
Approved By (PI) : Stephen Chamberland, Ph.D.
Date: 9/11/12
PI Signature: ___________________________________________________
Date: ______________
EH&S Signature: ________________________________________________
Date: ______________
PRINTED 2/9/2016
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KUHLKEN, CHEMISTRY DEPARTMENT, CENTRAL WASHINGTON UNIVERSITY
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