Gummy Cannabinoid Extraction Utilizing the Quecher's Method

Materials 

1. 50mL Centrifuge Tube
   
2. 10mL Pipette Tips
   
3. 0.5g Sodium Chloride - NaCL
4. 2.0g Magnesium Sulfate Anhydrous MgSO4
5. 10mLHPLC Grade Acetonitrile
6. 10mL HPLC Grade Water
7. 1g-1.5g Gummy Cannabinoid Sample
8. Laboratory Spatula/Spoon
9. Weigh Boats
10. Centrifuge Test Tubes

Equipment:

1. Vortex
2. Sonicator
3. Analytical Balance
4. 10mL Pipette
   
5. Centrifuge Tubes

Safety Issues

1. Disposable gloves and safety glasses should be worn at all times when handling chemicals. 
   
1. Acetonitrile is severely irritating to the eyes and slightly irritating to the skin.
   
1. Extended contact can lead to absorption and more irritation.
   
2. Refer to Methanol MSDS in the event of skin or eye contact or inhalation.

1. Take a gummy sample and weigh it on the analytical balance. Record exact mass in mg.
2. Place 1 g to 1.5g of gummy in a falcon tube.
3. Use the 10mL  pipette to transfer 10mL of HPLC grade water into the tube.
4. Cap and vortex for several minutes. Shape of the sample should begin to get lost.
5. Place in the sonicator at 45C for 15 minutes.
6. Vortex sample until it has dissolved.
7. Let the tube with sample equilibrate to room temperature.
8. Using a different pipette tip, Add 10mL of HPLC grade acetonitrile into the centrifuge tube.
9. Vortex sample for 1 minute until thoroughly mixed
10. Take the 0.5g bag of sodium chloride (NaCl) and add it to the centrifuge tube.
11. Take the 2.0g bag of magnesium sulfate (MgSO4) and add it to the centrifuge tube next.
12. Cap the centrifuge tube, vortex, and shake until salts are dissolved.
13. Place the centrifuge test tube in the centrifuge and turn on the centrifuge for 3 minutes. NOTE: Balance the centrifuge test tubes by placing each sample tube across from each other within the centrifuge. If only 1 sample is being prepared, counter-weigh the centrifuge by adding a 50mL tube containing enough water to be comparable to the volume of your sample.

    How to balance samples within a centrifuge
14. Remove test tubes, take top layer and filter 1mL through 0.22um filter into a green vial

1. Place the small vials in the sample prep holding tray, from left to right starting with the green vial, followed by the yellow, and then blue.
2. Use the 1 mL syringe to draw up 1 mL of extract solution containing sample
   
3. Place a syringe filter on the tip of the syringe.
   
4. Unscrew the green cap vial in the sample prep tray.
   
5. Dispense liquid in the syringe into the open green cap vial.
   
6. Discard the syringe and syringe filter, and put the green cap back on.

Note: Every vial that contains methanol should be tightly capped whenever it is not immediately in use. The caps of the vials should be removed only when they are being used to transfer liquid in between the vials in use. This will prevent evaporation of methanol and provide you with more accurate results. 

1. Vortex the vial briefly.
2. Set the micropipette to 40 uL and attach a clean tip.  
   
3. Prepare/prime the pipette tip for transfer by drawing up and dispensing the green cap solution three times into the green cap vial.
4. Transfer 40 uL from the green cap vial to the yellow cap vial.
   
5. Ensure that all of the solution has been transferred by drawing up and dispensing the yellow cap vial solution three times.
   
6. Cap both vials and shake or vortex the yellow vial well.
   
7. Discard the used pipette tip, attach a new, clean, one, and transfer 40 uL of the yellow vial solution to the blue cap vial.
   
8. Once again, ensure complete transfer by drawing up and dispensing the blue cap vial solution three times.
   
9. Cap both vials and shake or vortex the blue vial well.
   
10. The sample is now prepared and ready for HPLC Analysis.