Separation of Tetrahydrocannabinol Fraction from Cannabis Indica Extracts by Chromatographic Method

Cannabis indica contains a range of bioactive cannabinoids, among which tetrahydrocannabinol (Δ<sup>9</sup>-THC) is the principal psychoactive compound with wide pharmacological applications. Optimizing extraction efficiency and purity is essential for both analytical and therapeutic uses. This study aimed to compare the effectiveness of two solvent extraction techniques reflux condensation and long-term soaking for isolating Δ<sup>9</sup>-THC from C. indica leaves. Sample A was prepared using a reflux condenser to promote solvent recirculation and maintain elevated extraction temperatures, while Sample B was obtained by soaking the plant material in solvent for seven days under ambient conditions. The extracted oils were characterized using thin-layer chromatography (TLC), gas chromatography–mass spectrometry (GC–MS), infrared spectroscopy (IR), and high-performance liquid chromatography (HPLC). Physicochemical parameters including acid value, saponification value, specific gravity, and refractive index were determined through standard analytical methods. Additionally, qualitative assays were conducted to detect proteins, carbohydrates, steroids, tannins, gums, and mucilage. Comparative analysis revealed that the reflux-assisted extraction produced a higher yield and greater purity of Δ<sup>9</sup>-THC, with improved physicochemical stability compared to the soaking method. These results indicate that reflux condensation offers a more efficient and reproducible approach for cannabinoid extraction. The findings contribute to refining extraction methodologies and enhancing the quality of cannabinoid-based research and pharmaceutical formulations.