Gummy Sample Preparation Using the Quechers Method - HPLC Hemp Analysis

How to Prepare Gummy Samples for HPLC Analysis

Gummy Cannabinoid Extraction 




















This cannabinoid extraction process utilizes the QuEChERS (quick, easy, cheap, effective, rugged, and safe) method. The two solvents used will be acetonitrile and water. The salts used to aid in the extraction of cannabinoids from the gummy consists of sodium chloride and magnesium sulfate anhydrous.

This procedure is compatible with the GummyPak line of products:

Integration Kit:
GummyPak Starter - Integration kit required to use the GummyPak Pre-packaged consumables. Includes a 50mL centrifuge, a 1-10mL pipette, and a GummyPak Novice kit. 

Consumables:
GummyPak Novice - All consumables needed to prepare 25 Gummy samples utilizing the QuEChERS method. Salts are Pre-packaged for ease of use. Centrifuge and scale not included.

GummyPak Pro - All consumables needed to prepare 50 Gummy samples utilizing the QuEChERS method. Salts are Pre-packaged for ease of use. Centrifuge and scale also not included.

Materials 
  • 50 mL Centrifuge Tubes
  • 10 mL Pipette Tips
  • 0.5 g Sodium Chloride (NaCL)
  • 2.0 g Magnesium Sulfate Anhydrous (MgSO₄)
  • 10 mL HPLC Grade Acetonitrile
  • 10 mL HPLC Grade Water
  • 1 g - 1.5 g Gummy Cannabinoid Sample
  • Laboratory Spatula / Spoon
  • Weigh Boats

Equipment:

  • Vortex
  • Sonicator
  • Analytical Balance
  • 10 mL Pipette
  • Centrifuge Tubes
  • Centrifuge

Safety Issues

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

Instructions

  1. Place an empty weigh boat on the balance and tare it to 0.
  2. Take a whole gummy, weigh it on the analytical balance, and record weight of the entire piece.
    1. This weight will be used to determine potency in units of mg/gummy
  3. Place a centrifuge tube on the balance and tare it to 0.
  4. Weigh roughly 1000 mg of the gummy in a centrifuge tube. Record weight onto the sample tube.
  5. Use the 10 mL pipette to transfer 10 mL of HPLC grade water into the tube.
  6. Cap and vortex for several minutes. The sample should begin to lose its' shape.
  7. Place in the sonicator at sonicate the sample at 45C for 15 minutes.
  8. Vortex sample until it has dissolved.
  9. Let the tube with sample equilibrate to room temperature.
  10. Using a different pipette tip, Add 10 mL of HPLC grade acetonitrile into the centrifuge tube.
  11. Vortex sample for 1 minute until thoroughly mixed
  12. Open the bag of salts, sodium chloride (NaCl) and magnesium sulfate (MgSO₄), and add the salts to the centrifuge tube.
  13. Cap the centrifuge tube, vortex, and shake until salts are dissolved. Note: This process generates heat.
  14. Place the centrifuge test tube in the centrifuge and turn on the centrifuge for 3 minutes. NOTE: Unbalanced centrifuges may cause damage and can potentially injure the operator.
    1. Do not open the centrifuge door while it's moving.
    2. Balance the weight distribution of the centrifuge by placing each sample tube across from each other within the centrifuge.
    3. If only 1 sample is being prepared, counter-weigh the centrifuge by adding a 50 mL tube containing enough water to be comparable to the volume of your sample

How to balance samples within a centrifuge
  1. After the centrifuge has been run for 3 minutes, remove test tubes, Use a new syringe to draw 1 mL of the top, organic, portion from the solution.
  2. Place a 0.22 micron filter on the fully drawn syringe and dispense the 1 mL solution through the filter and into a green-capped, 2 mL vial.

Gummy Sample Dilution Process - For analysis with Cann-ID

Materials

  • 200 uL Pipette Tips

Equipment

  • 10 uL -100 uL Pipette
  • Vortex

Instructions

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

The sample is now prepared and ready for HPLC analysis.

Documentation on Hazardous Chemicals

  • Methanol MSDS

  • Acetonitrile MSDS

  • Sodium Chloride MSDS

  • Magnesium Sulfate Anhydrous MSDS




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