This protocol demonstrates the whole process of CBD extraction, starting from preparing the material, solvent extraction and measuring CBD content. By suggesting the optimized solvent and solvent concentration, this method eliminates unnecessary processes and can help in efficient solvent extraction for industrial purposes. Begin by obtaining Cherry Wine inflorescences from plants grown in the field.
Air dry the inflorescences at 35 degrees Celsius for 48 hours. Grind the inflorescences using a grinding machine set at 177 micrometers. Pass the pulverized material through number 80 mesh sieve and store the powder in a sealed bag at room temperature.
In a 50-milliliter conical tube, weigh 0.5 grams of the cannabis inflorescences powder. Add 40 milliliters of 50%ethanol prepared in deionized water. Place the extraction vessel in an ultrasonic bath set at 40 kilohertz set room temperature.
Perform the extraction for 30 minutes, increasing the temperature of the bath from 25 to 30 degrees Celsius. After sonication, decant the extraction fluid into a centrifuge tube. Centrifuge the fluid at 3000-times G at 15 degrees Celsius for 15 minutes.
Filter the supernatant through an 8-micrometer filter paper under vacuum. Dilute the cannabinoid standards to operating concentrations of 100, 50, 25, and 12.5 micrograms per milliliter in 100%methanol. Mix and sonicate for five minutes in an ultrasonic bath set at 40 kilohertz and sonication power of 100 watts.
Filter the standards and the cannabis sample supernatant through a 0.45-micrometer polytetrafluoroethylene syringe filter. Put the samples in 1.5 milliliter vials, and place them into the HPLC autosampler. Load 10 microliters of the sample and run the HPLC using the parameters shown in this table.
Generate a standard curve and derive cannabinoid concentrations in 50 to 200 micrograms per milliliter. Calculate the weight of cannabinoids in micrograms by multiplying with the volume of the solvent used in the extraction process, convert the value to milligrams by dividing it by 1, 000. Divide this value by the weight of the plant material used in the extraction to obtain milligrams per gram of dry weight.
HPLC analysis showed that when using 100%solvents, acetonitrile was most favorable for the extraction of individual cannabinoids. However, ethanol was further examined due to the lowest toxicity of all. Evaluation of the impact of aqueous ethanol on concentration of extracted cannabinoids showed that maximum extraction was observed at 50%ethanol.
There was a 39.7%increase over absolute ethanol for the extraction of CBDA. The level of THCA was also reduced by 20.3%Using the design-of-experiment approach, extraction of CBDA plus CBD was optimized. Response surface methodology analysis confirmed the ideal parameters as 30-minute extraction, 53.4%ethanol in water, and 1 to 100 sample-to-solvent ratio.
A higher amount of CBDA was obtained using the ultrasonic-assisted extraction method compared to the maceration method. The amount of extracted CBD also doubled using the ultrasonic method, indicating higher efficiency for cannabinoid extraction. It is important to remember the sample to solvent ratio, extraction time and solvent concentration.
optimized by this study. This procedure can be performed for extraction of other cannabinoids from hemp.
