This study analyzed Tuber melanosporum, Tuber aestivum,and Tuber magnatum, specifically the content of β-glucan, amino acids, nucleic acid-related substances, vitamin C, and ergosterol. The β-glucan content was highest in T. magnatum (20.54%). The free amino acid content was highest in T. aestivum, with the major amino acids being cystine, glutamic acid, and aspartic acid. The total amino acid content was the highest in T. magnatum, the primary amino acids being cystine, glutamic acid, and lysine. The content of nucleotide related compounds was the highest in T. magnatum (5.09 mg/100 g). The highest content of vitamin C (10.15 mg/100 g) and ergosterol (596.91 mg/100 g) was found in T. magnatum, which had the highest useful component content among the three truffles. Additional studies investigating the variation in useful components according to collection time and growth environment are needed.