I would appreciate it if anyone could share with me the protocol to extract cannabinoids for testing the total Δ9-THC content in fish tissue or feed. Thanks
Idowu Goodnews Oluwaseyi
Homogenization:Weigh approximately 1-5 grams of fish tissue or feed sample. Homogenize the sample using a mortar and pestle or an appropriate homogenizer with a small amount of ethanol to aid the process. Extraction:Transfer the homogenized sample to a glass flask. Add 20 mL of ethanol and mix thoroughly. Add 10 mL of water and mix again. Add 5 g of sodium chloride to the mixture to enhance the extraction process. Vortex or shake the mixture for 10-15 minutes to ensure thorough mixing. Separation:Add 30 mL of hexane to the mixture and shake or vortex vigorously for 10 minutes. Centrifuge the mixture at 3000 rpm for 10 minutes to separate the phases. Collect the upper hexane layer (organic phase) containing the extracted cannabinoids. Drying:Add anhydrous sodium sulfate to the collected hexane layer to remove any residual water. Swirl the mixture and allow it to stand for a few minutes. Filter the hexane layer through a fine filter to remove the sodium sulfate. Concentration:Transfer the hexane extract to a rotary evaporator flask. Evaporate the hexane under reduced pressure at a temperature not exceeding 40°C to concentrate the extract. If necessary, dry the final residue under a gentle stream of nitrogen gas to remove any remaining solvent. Preparation of Sample for HPLC or GC-MS:Dissolve the dried residue in a small volume of methanol (e.g., 1-2 mL). Filter the solution through a 0.22 µm filter to remove any particulate matter. Calibration and Standard Preparation:Prepare a series of Δ9-THC standard solutions in methanol at known concentrations for calibration purposes. Run the standards through the HPLC or GC-MS system to create a calibration curve. Sample Injection and Analysis:Inject the filtered sample extract into the HPLC or GC-MS system. Use a suitable column and detector settings optimized for cannabinoid analysis. Commonly, a C18 column with UV detection or a suitable MS detector is used. Analyze the chromatograms and quantify the Δ9-THC content by comparing the sample peaks to the calibration curve.
Sure, here is a detailed protocol for extracting cannabinoids, specifically Δ9-tetrahydrocannabinol (Δ9-THC), from fish tissue or feed for testing purposes:
Materials Needed:
- Fish tissue or feed sample
- Ethanol (analytical grade)
- Methanol (analytical grade)
- Water (HPLC grade)
- Sodium chloride (NaCl)
- Anhydrous sodium sulfate (Na2SO4)
- Hexane
- Chloroform
- Glassware (beakers, flasks, tubes, etc.)
- Mortar and pestle or homogenizer
- Centrifuge
- Rotary evaporator
- Nitrogen gas (for drying)
- HPLC or GC-MS system with appropriate columns and detectors
- Standard solutions of Δ9-THC for calibration
Sample Preparation:
Analysis:
Notes:
- Ensure all solvents and reagents used are of analytical or HPLC grade to avoid contamination.
- Maintain consistent temperature and handling conditions for all samples and standards.
- Use proper safety equipment and protocols when handling organic solvents and biological samples.
By following this protocol, you should be able to efficiently extract and quantify the total Δ9-THC content in fish tissue or feed samples.
NAIJA TO THE WORLD!
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