3.2.2. Dynamic volatile collection

8. Briefly disconnect the vacuum line once the bees have become relatively calm.

9. Place a protective screen (Teflon sheath or screen mesh) around the tip of an adsorbent (e.g. Porapak Q, Super Q or Tenax) filter to protect it from being blocked with wax by the bees.

10. Place the filter at the vacuum line’s tip and reconnect it to the lid of the container.

Avoid placing the vacuum supply source such as a pump and odour source on the same bench to prevent alarming the bees from the vibrations/noise from the pump.

11. Pull air out of the system at a desired flow rate and for a specific duration (see section 2.2.2. and 2.2.3. on in situ volatile collection and analysis). The sucrose solution serves a dual role; as a source of food to keep bees alive and to calm them.

12. Remove the filter and seal the ends with Teflon tape after volatile collection is complete.

13. Switch-off the vacuum.

14. Disassemble the odour container.

15. Return the bees to their colony of origin.

16. Elute the volatiles adsorbed on the filter using an appropriate volume of solvent (e.g. 200 µl) such as dichloromethane or redistilled ether in to a sample vial using a gentle flow of N2 gas.

17. Label the sample bottle and store at -80°C prior to analysis.