B Odour Mixing Protocol - michelletytong/TRGProtocols GitHub Wiki
Written by Arish Mudra Rakshasa (Sept 2017) Earlham College, Richmond, IN, USA
Preparation
Wear clean gloves and a lab coat, and leave all personal items outside the lab. Gather the following materials:
- Clean vials and caps (as many as there are odours to be mixed).
- P20, P200, and P1000 micropipettes.
- P20, P200, and P1000 micropipette filter tips.
- Clean 100 mL graduated cylinder.
- Mineral oil (open new and label with date, if needed).
- Required odourants (found in the flammables cabinet in SH 161).
- Mixture amount reference sheet (Talk to Michelle Tong)
- Typical odourants: (+)-limonene (Cat no: 183164 Sigma-Aldrich) and (-)-limonene (Cat no: 218367 Sigma-Aldrich)
Procedure
Mineral oil
-
Transfer 50 mL of mineral oil into a graduated cylinder. Precautions: mineral oil is viscous, so pour along the sides of the cylinder to avoid air bubbles. Pour slowly to avoid overshooting, because it takes a little bit for it to settle. Observe the lower meniscus at eye level to get an accurate measurement.
-
Label a clean vial “MO” along with your initials and the date using tape.
-
Transfer the 50 mL mineral oil into this vial.
Precautions: again, due to viscosity of the oil, pour it slowly and along the sides of the vial, to avoid air bubbles. Try to transfer as much of it as possible.
Odourant
-
Transfer 50 mL of mineral oil into a graduated cylinder.
-
Label a clean vial with the name of the odourant, your initials, and the date.
-
Transfer the 50 mL mineral oil into this vial.
-
Find the appropriate amount of odourant to create a solution of approximately 1 Pa vapour pressure (ref. Odour_Dilutions_TONG.xls). For (+)/(-)limonene, this is 102 µL.
Precautions: only use filter tips to transfer odourants!
- Using the appropriate micropipette, transfer this amount of odourant into the vial of mineral oil. For limonene, you would use a P200 set to 102 µL.
Precautions: feel free to immerse your pipette tip into the mineral oil to get all the odourant out. Do not tilt your pipette too much.
- Secure the cap on the vial and invert the vial several times to thoroughly mix the odourant with the mineral oil. Set the prepared vials in a tray or box. Avoid carrying them in your hands.
Fresh odours have to be mixed every day an experiment is performed. Odours from the previous day cannot be reused!
Clean-Up
- Clean all glassware in the sinks and leave them to dry.
- Make sure that there are enough pipette tips, clean vials, and chemicals for the next time someone needs to mix odours.
- Wipe down the workspace with alcohol.