| Ophiopogon japonicus(L.f.) Ker-Gawl is an evergreen perennial plant, widely distributed in Southeast Asia. In China, the tuber of O. japonicus has been used as the traditional Chinese medicine for thousand years, and also can be used as health food raw materials. As a traditional Chinese medicine, it has been used to cure acute and chronic inflammation and cardiovascular diseases. Previous studies have revealed that it rich in bioactive components such as saponins, homoisoflavonoids and polysaccharides. In recent years, researches on O. japonicus tuber were focused on the separation and purification of the monomer whith have better pharmacologically active. As a health food raw material, O. japonicus tuber has high nutritional and health value, but there is few report about that. According to the literature, various extracts of O. japonicus tuber also have obvious pharmacological effects, these pharmacological effects may be due to a variety of active ingredients extract synergies result, however, the chemical composition of the extract has been reported rarely. In this study, the active ingredient of O. japonicus tuber extract was isolated and purified, and the chemical composition was analyzed by HPLC-MS and GC-MS methods, while O. japonicus tuber nutrients and its in vitro biological activities were evaluated, the specific contents are as follows:1. Four compounds were isolated from the extracts of O. japonicus tuber and were identified by the information of ESI-MS, 1H-NMR and 13C-NMR. The compounds were known compounds methylophiopogonanone A, methylophiopogonanone B, ophiopogonin D and ophiopogonin D’.2. The chemical composition of O. japonicus tuber extracts were analyzed by HPLC-MS and GC-MS. 17 homoisoflavonoid compounds were identified in the chloroform/methanol extract by HPLC-MS. 34 compounds were identified in four different extracts by GC-MS.3. O. japonicus tuber nutrients were measured, including proximate nutrients(ash, moisture, crude fat, crude fiber, crude protein and carbohydrates), the content of mineral elements(K, Na, Ca, Mg, Cu, Fe, Zn, Mn), amino acid composition and fatty acid content. The results showed that the carbohydrate and crude protein content were higher in O. japonicus tuber.The content of K, Ca and Mg were higher among 8 mineral. 16 kinds ofamino acids were also detected, the content of glutamic acid was the most highest. Essential amino acid content was 33.72% of the total. 5 fatty acids were detected in O. japonicus tuber. They were palmitic acid, seventeen acid, linoleic acid, linolenic acid and stearic acid. Linoleic acid is an unsaturated fatty acid, and its content was the highest.4. The active ingredients of O. japonicus tuber were measured, including total phenolic and flavonoid content. The content of methylophiopogonanone A and methylophiopogonanone B of the extracts were determined by HPLC-DAD method. The content of ophiopogonin D and ophiopogonin D’ in different extracts were determined by HPLC-ELSD. Among the extracts, the total phenols and flavonoids of the n-butanol extract were the highest, and the methylophiopogonanone A, methylophiopogonanone B, ophiopogonin D and ophiopogonin D’ contents in the n-butanol extract were also the highest. The result indicating that n-butanol is the effective extraction solvent of the 4 compounds.5. The in vitro activities of O. japonicus tuber including in vitro antioxidant,the enzyme inhibitory and antibacterial activities were evaluated. Antioxidant measurement results were shown that the chloroform/methanol and n-butanol extract have good antioxidant activity, while the crude polysaccharides has lower antioxidant activity. The correlation coefficient analysis were shown that the antioxidant activity of the extracts were significantly correlated with their total phenols and flavonoids contents. In the experiment of enzyme inhibition activity, the α-glucosidase, α-amylase and lipase inhibitory activities of the extracts and monomer compounds were evaluated. The results showed that the extracts and monomer compounds have no α-amylase and lipase inhibitory activities. The n-butanol and chloroform/methanol extract showed strong α-glucosidase inhibitory activity, and α-glucosidase inhibition rate of the n-butanol extract was higher than acarbose. The in vitro antibacterial activity of the extracts and monomer compounds against Staphylococcus aureus CMCC26003, Bacillus subtilis ATCC6633, Bacillus cereus. Frankland CMCC(B)63301, Staphylococcus epidermidis CMCC(B)26069, Escherichia coli CMCC44102 and Enterobacter aerogenes GIM1.108 were evaluated. The results showed that the chloroform/methanol extract and methylophiopogonanone A have strong in vitro antibacterial activity against Bacillus subtilis ATCC6633 and Bacillus cereus. Frankland CMCC(B)63301. |
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