The purpose of this study is to elucidate antioxidative characteristics and the active compounds of Wonchuri (Hemerocallis fulva L.)
First, the content of general components contained in the freeze dried H. fulva L., and reducing sugar, free sugar, free amino acids, free fatty acid, antioxidant ...
The purpose of this study is to elucidate antioxidative characteristics and the active compounds of Wonchuri (Hemerocallis fulva L.)
First, the content of general components contained in the freeze dried H. fulva L., and reducing sugar, free sugar, free amino acids, free fatty acid, antioxidant vitamins, organic acid and mineral were quantified.
In vivo experiments, weanling male Sprague-Dawley rats were fed normal, control, 5% HFP (H. fulva L. powder) and 10% HFP diet for 4 weeks and intoxicated with 10% ethanol in drinking water during the last two weeks of the experiment.
The 80% ethanol extract of Wonchuri was fractionated sequentially with hexane, chloroform, ethyl acetate and butanol. Each fraction was studied for antioxidative chracteristics in vitro. To determine antioxidative characteristics, radical scavenging activities, inhibitory effects on linoleic acid autoxidative, metal-chelating effects, reducing power, SOD-like activity were measured.
To separate an antioxidative compounds, ethyl acetate and butanol fractions showing antioxidative capacity in several experiments were subjected to column chromatography on silica gel and prep-HPLC. The structure of the isolated compounds was identified by UV/vis, IR, FAB-MS and NMR spectrum analysis.
The results was stated below.
1. The carbohydrate, crude protein, crude lipid and crude ash contents were 54.56%, 24.35%, 3.44% and 7.23%, respectively. The amount of reducing sugar was 17.18% and that of free sugar was 1,897.68 mg%. The main components of free sugar was fructose.
The contents of essential amino acids was 159 mg% and the major amino acids was glutamic acid (638 mg%). The major fatty acids was ?-linolenic acid and the ratio of unsaturated fatty acid to saturated fatty acid was 3.86. The unsaturated fatty acid contents of sample was 3 times higher than that of saturated fatty acid contents. Lactic acid was the major organic acids in sample. The contents of total ascorbic acid and dihydro ascorbic acid were 278.16 mg% and 112.84 mg%. The K, P, Ca and Mg levels were greater than those of other minerals, with K comprising more than 81.90% of the total mineral content.
2. The content of MDA (in the liver tissues), plasma total cholesterol, and plasma triglyceride of HFP group was significantly decreased compared with control group (p<0.05).
Alcohol intoxication increased SOD activity of liver compared with activities of catalase and GPx too great extent. Activities of SOD, GPx and catalase of liver of HFP group were lower than those of control group. But the GPx activity was not significant.
3. The contents of total phenol in ethyl acetate, chloroform, butanol, hexane and aqueous fraction were 749, 226, 205, 63 and 25 mg/g, respectively. The contents of total flavonoid in ethyl acetate, butanol, chloroform, aqueous and hexane fraction were 323, 67, 36, 29 and 17 mg/g, respectively.
4. Hexane and chloroform fraction inhibited linoleic acid autoxidation while radical scavenging activities were prominent in ethyl acetate fraction. In butanol fraction, DPPH, nitrite and hydroxyl radical showed great activity. The fractions of 80% ethanol extract were higher chelating capacity for Fe3+ than Cu2+. At FeCl3 of 10-4 M, all the fractions except for aqueous fraction recorded 65% or higher chelating effects. At CuSO4 of 10-4 M and 10-3 M, butanol fraction showed greater chelating effects than the other fractions. Reducing power was the highest in the order of ethyl acetate, butanol, chloroform, and hexane and aqueous fraction.
The oxidative stress improvement effects of Wonchuri were also examined with LLC-PK1 cells. As a result, it had protective effects on oxidative stress caused by LOO?, NO, O2? and ONOO? with the effects recording the highest level in ethyl acetate fraction, where cell viability was significantly increased.
5. The isolation and purification process applied to ethyl acetate and butanol fraction of Wonchuri led to the isolation of isoquercitrin, isovitexin, physcion, quercitrin, ethyl ferulate, caffeic acid, ?-amyrin acetate and chlorogenic acid.
6. Of the isolated compounds, isoquercitrin, chlorogenic acid, and standard ?-tocopherol and vitamin C were examined for synergic effects in the linoleic acid system for 7 days. As a result, antioxidant effects were the greatest when the four compounds worked together.
From the above result, Wonchuri (Hemerocallis fulva L.) could be a good physiological food since it is a good source vitamin C and A, ?-tocopherol, organic acid, and antioxidants such as phenolic compounds and has excellent antioxidant effects in vivo and in vitro. When they are researched and examined in more multifaceted ways, they are highly likely to make a contribution to the prevention and treatment of various chronic diseases caused by oxidative stress.
The purpose of this study is to elucidate antioxidative characteristics and the active compounds of Wonchuri (Hemerocallis fulva L.)
First, the content of general components contained in the freeze dried H. fulva L., and reducing sugar, free sugar, free amino acids, free fatty acid, antioxidant vitamins, organic acid and mineral were quantified.
In vivo experiments, weanling male Sprague-Dawley rats were fed normal, control, 5% HFP (H. fulva L. powder) and 10% HFP diet for 4 weeks and intoxicated with 10% ethanol in drinking water during the last two weeks of the experiment.
The 80% ethanol extract of Wonchuri was fractionated sequentially with hexane, chloroform, ethyl acetate and butanol. Each fraction was studied for antioxidative chracteristics in vitro. To determine antioxidative characteristics, radical scavenging activities, inhibitory effects on linoleic acid autoxidative, metal-chelating effects, reducing power, SOD-like activity were measured.
To separate an antioxidative compounds, ethyl acetate and butanol fractions showing antioxidative capacity in several experiments were subjected to column chromatography on silica gel and prep-HPLC. The structure of the isolated compounds was identified by UV/vis, IR, FAB-MS and NMR spectrum analysis.
The results was stated below.
1. The carbohydrate, crude protein, crude lipid and crude ash contents were 54.56%, 24.35%, 3.44% and 7.23%, respectively. The amount of reducing sugar was 17.18% and that of free sugar was 1,897.68 mg%. The main components of free sugar was fructose.
The contents of essential amino acids was 159 mg% and the major amino acids was glutamic acid (638 mg%). The major fatty acids was ?-linolenic acid and the ratio of unsaturated fatty acid to saturated fatty acid was 3.86. The unsaturated fatty acid contents of sample was 3 times higher than that of saturated fatty acid contents. Lactic acid was the major organic acids in sample. The contents of total ascorbic acid and dihydro ascorbic acid were 278.16 mg% and 112.84 mg%. The K, P, Ca and Mg levels were greater than those of other minerals, with K comprising more than 81.90% of the total mineral content.
2. The content of MDA (in the liver tissues), plasma total cholesterol, and plasma triglyceride of HFP group was significantly decreased compared with control group (p<0.05).
Alcohol intoxication increased SOD activity of liver compared with activities of catalase and GPx too great extent. Activities of SOD, GPx and catalase of liver of HFP group were lower than those of control group. But the GPx activity was not significant.
3. The contents of total phenol in ethyl acetate, chloroform, butanol, hexane and aqueous fraction were 749, 226, 205, 63 and 25 mg/g, respectively. The contents of total flavonoid in ethyl acetate, butanol, chloroform, aqueous and hexane fraction were 323, 67, 36, 29 and 17 mg/g, respectively.
4. Hexane and chloroform fraction inhibited linoleic acid autoxidation while radical scavenging activities were prominent in ethyl acetate fraction. In butanol fraction, DPPH, nitrite and hydroxyl radical showed great activity. The fractions of 80% ethanol extract were higher chelating capacity for Fe3+ than Cu2+. At FeCl3 of 10-4 M, all the fractions except for aqueous fraction recorded 65% or higher chelating effects. At CuSO4 of 10-4 M and 10-3 M, butanol fraction showed greater chelating effects than the other fractions. Reducing power was the highest in the order of ethyl acetate, butanol, chloroform, and hexane and aqueous fraction.
The oxidative stress improvement effects of Wonchuri were also examined with LLC-PK1 cells. As a result, it had protective effects on oxidative stress caused by LOO?, NO, O2? and ONOO? with the effects recording the highest level in ethyl acetate fraction, where cell viability was significantly increased.
5. The isolation and purification process applied to ethyl acetate and butanol fraction of Wonchuri led to the isolation of isoquercitrin, isovitexin, physcion, quercitrin, ethyl ferulate, caffeic acid, ?-amyrin acetate and chlorogenic acid.
6. Of the isolated compounds, isoquercitrin, chlorogenic acid, and standard ?-tocopherol and vitamin C were examined for synergic effects in the linoleic acid system for 7 days. As a result, antioxidant effects were the greatest when the four compounds worked together.
From the above result, Wonchuri (Hemerocallis fulva L.) could be a good physiological food since it is a good source vitamin C and A, ?-tocopherol, organic acid, and antioxidants such as phenolic compounds and has excellent antioxidant effects in vivo and in vitro. When they are researched and examined in more multifaceted ways, they are highly likely to make a contribution to the prevention and treatment of various chronic diseases caused by oxidative stress.
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