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Mushrooms are considered as functional foods worldwide not only for their flavor and nutritional properties but also for their pharmacological potential. They are rich in proteins, minerals, carbohydrates, vitamins, fibers as well as containing a significant amount of phenolic compounds and essential fatty acids. Mushrooms are a significant source of essential fatty acids for a human healthy diet. These essential fatty acids include linoleic acid ranging between 0–87%, oleic acid between 0–65%, and linolenic acid between 0–18%. Fatty acids, particularly polyunsaturated ones, omega-3 and -6 series, are necessary for the treatment of and preventing hypertension, cancer, diabetes, coronary artery disease, arthritis, and other inflammatory, osteoporosis, and autoimmune disorders. Generally, fatty acid contents are analyzing by GC and GC-MS. Fatty acids need to be derivatized to increase their volatility. Fatty acid methyl esters are prepared by different methylation techniques and then separated on appropriate columns and identified by a flame ionization detector (FID). This chapter focuses on fatty acid contents measured in mushroom species over the last two decades. Also, the importance of linoleic, oleic, linolenic, palmitic, and stearic acids, which are the main fatty acids, and the significance of ω-6/ω-3 ratio in the human diet are explained. Furthermore, fatty acids isolated from mushrooms and their bioactivities are also reported in this review.

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