Cudrania tricuspidata has been used as an important traditional herbal medicine for anti-tumor, anti-oxidative, hepatoprotective, gastritis, and anti-inflammation in East Asian countries such as Korea, China, and Japan. The fruit of Cudrania tricuspidata (CT fruits) has been used to make fresh juice, jam, wine and vinegar in Korea. Nevertheless, there are no sufficient data on chemical constituents and their biological activities of CT fruits compared with leaf, bark and root. Therefore, this studies were carried out to investigate new bioactive components that had not been previously reported in CT fruits, and changes of bioactive and nutritional components during fermentation of CT fruits by Armillaria mellea. The results were as follows.

1. Three new phenolic compounds have been isolated from CT fruits by silica gel, Sephadex LH-20 and flash column chromatography on RP C18 cartridge. These compounds were identified as 4-hydroxybenzyl alcohol and 4-(β-D-glucopyranosyloxy)benzyl alcohol (gastrodin) by UV profiles, GC-MS, LC-MS, 1H-NMR and 13C-NMR, respectively, and tris[4-(β-D-glucopyranosyloxy)benzyl] citrate (parishin A) by UV profiles, GC-MS, LC-MS, and 1H-NMR, 13C-NMR, DEPT, 2D-NMR (COSY, HMBC, HMQC) spectrometry. In addition, parishin B, parishin C and parishin E have been tentatively identified by UV profile and comparisons of HPLC retention times with those of authentic compounds, These components have been known as major pharmacological compounds of Gastrodia elata rhizome, but were firstly identified from C. tricuspidata in present study.

2. The content of 4-hydroxybenzyl alcohol, gastrodin, parishin B, parishin A, parishin C and parishin E were generally higher in bark and root than in leaf, seed and twig, and in immature fruits than in mature and over-mature fruits.

3. The selection of ethanol and water used as extraction solvent in this study because they were safer and less toxic as compare to other organic solvent. Ethanol concentration as extraction solvent had significant effect on contents of parishin derivatives, phenolic acids, flavonoids, and antioxidant capacities (DPPH and ABTS) in extracts. The effect of ethanol concentration on parishin derivatives, total phenol, total flavonoid, content of phenolic acid, DPPH ABTS followed parabola shape, in which the highest value for all assays were at 60-80% ethanol except ABTS at 80% ethanol. In contrast, the ABTS was increased as the ethanol concentration was increased, reaching a maximum ABTS at 100% ethanol concentration.

4. Methods for determining parishin derivatives in C. tricuspidata by HPLC method were validated by determining linearity, limit of detection (LOD), limit of quantitation (LOQ), recovery, precision, intraday and interday reliability. Linearity, limit of detection (LOD), limit of quantitation (LOQ). The results showed that high linearity with a coefficient of correlation (R2) were ranged from 0.9977 to 0.9987 for 5 compounds, 96-106% for recovery, 0.0648-0.2410 μg/mL for LOD, 0.0749-1.1149 μg/mL for LOQ. respectively.

5. Soluble solid, free sugar, free amino acid and carotenoid pigments were decreased during fermentation while changes of pH and total amino acids did not observed even after 10 day fermentation.

6. Content of parishin derivatives were decreased during the 10-day fermentation period. the crude enzyme preparation from cultured mycelia of A. mellea was found to convert parishin A into 4-hydroxybenzyl alcohol via parishin B and parishin C with high conversion ratio, The enzymatic hydrolysis pathway of parishin A by A. mellea enzyme was parishin A → parishin B + parishin C → gastrodin → 4-hydroxybenzyl alcohol.