Psilocybin mushrooms are magic or hallucinogenic mushrooms that consist of psilocybin, which is converted into psilocin upon digestion. This compound is widely used as a recreational drug, but, depending on dosage, psilocybin can cause acute and panic responses such as euphoria, changes in perception, perceived spiritual experiences, and feelings of detachment. Besides psilocin, baeocystin, norlpsilocin, aeruginascin, and norbaeocystin show psychoactive effects. These drugs are restricted for use in many countries. Nevertheless, with the regulatory framework, these psychotropic alkaloids are used in medications for treating mental illness, anxiety, and depression. However, the extraction of psychotropic compounds from psilocybin mushrooms is very difficult due to their complex chemical nature. In general, four major methods are followed, viz., liquid–liquid extraction, microwave-assisted extraction, ultrasound-assisted extraction (UAE), and high-performance liquid chromatography. Among them, the UAE method is highly reliable for extraction. The materials required, equipment used, and procedure followed are briefly discussed in this chapter.