# An Optimized Terpene Profile for a New Medical Cannabis Oil
## 3 Results  
### Extraction Methods for Medical Cannabis Oils  
In this work, two methods previously described in the literature for MCO preparation were applied [30; 32]. They included a decarboxylation step at 115 \({}^{\circ}\)C for 40 min and 130 \({}^{\circ}\)C for 30 min, respectively. In the first method, the plant material was extracted by maceration at 100 \({}^{\circ}\)C for 40 min [32] whereas in the second method, the extraction was carried out for 30 min by sonication at controlled temperature [30]. MCT oil was shown to extract and preserve a significant concentration of terpenes compared to olive oil [29]. For this reason, we chose to use MCT oil for all methods in order to minimise differences due to matrix effects. In addition to these methods, one novel extraction procedure (CERFIT method) was developed in this work. The plant material was stored at \(-\)20 \({}^{\circ}\)C until use; therefore, it was put in a fan oven at 25 \({}^{\circ}\)C for 60 min before weighing. The decarboxylation of the plant material was carried out at a lower temperature (100 \({}^{\circ}\)C) for a shorter time (10 min) in CERFIT than in the other two methods: this step was repeated five times in order to alternate heating and cooling to preserve the phytocomplex. Moreover, the closed flask was kept under controlled vacuum to avoid the loss of volatile compounds. At the end of the cyclic process, the flask was maintained at \(-\)25 \({}^{\circ}\)C for 60 min recovering essential oil volatile components. Then, the extraction was performed by placing a probe sonicator in the flask (MCT oil plus Cannabis flos) and the system in a refrigerated water bath. Finally, the product was filtered and packaged.  
Ten cannabis samples were extracted with each method and the experimental conditions are compared in Table 6. Starting from 5 g of cannabis flos in 50 mL of MCT oil, a final product of 50 mL of medical cannabis oil was obtained. Cannabinoid and terpene profile was analysed for all of the samples (10 samples for each method).  
\begin{table}
\begin{tabular}{c c c c} \hline \hline
**METHOD** & **SIFAP**[32] & **CALVI[30]** & **CERFIT** \\  & & & **Present Work** \\ \hline Cannabis variety (5 g)/(T) & Bedrocan (RT) & Bedrocan (RT) & Bedrocan (\(-\)20 \({}^{\circ}\)C) \\ \hline Matrix (50 mL) & MCT oil \({}^{\text{a}}\) & MCT oil \({}^{\text{a}}\) & MCT oil \\ \hline Cannabis/Matrix ratio & 1:10 & 1:10 & 1:10 \\ \hline Thawing & No & No & Yes/25 \({}^{\circ}\)C, 60 min fan oven \\ \hline Decarboxylation step & Yes/115 \({}^{\circ}\)C, 40 min & Yes/135 \({}^{\circ}\)C, 30 min & Yes/cyclic T \({}^{\text{b}}\), 8 h \\  & static oven & fan oven & vacuum microwave extractor \\ \hline Freezing & No & No & Yes/\(-\)20 \({}^{\circ}\)C, 60 min \\ \hline Extraction process & Turbo emulsion & Turbo emulsion & Turbo emulsion \\  & water bath (100 \({}^{\circ}\)C 40 min) & bath sonicator (35 KHz 30 min) & probe sonicator (200 W/30 KHz 8 min) \\ \hline Oil heating step & Yes & No \({}^{\text{c}}\) & No \({}^{\text{d}}\) \\ \hline Filtration & Yes/filter paper & Yes/filter paper & Yes/filter paper \\ \hline Preparation time (hour) & 2 & 1.5 & 9.5 (8 nightlong) \\ \hline \hline \end{tabular}  
* \({}^{\text{a}}\) modified from the original work where the used solvent was olive oil Ph. \({}^{\text{b}}\) cycles as described in Material and methods. \({}^{\text{c}}\) final temperature of the water in bath sonicator was 45 \({}^{\circ}\)C. \({}^{\text{d}}\) final temperature of the oil flask was 15 \({}^{\circ}\)C. T: temperature; CT: cyclic temperature; RT: room temperature.  
\end{table}
Table 6: Detailed comparison of the three methods used for Cannabis oil preparation in this study.