The Research About Antioxidant Activities Of The Functional Food Comman Material In China

With the development of economy and people’s living standards, food consumption is moving from subsistence to a comfortable level, people put a higher quality of food requirements; functional food thus came into being.Regulatory function of the functional food depends on its raw material and ingredients, but the functional food raw materials has been the lack of systematic research.China’s functional food is functionalse on the traditional Chinese medicine and Medicinal and Edible theories, the raw materials mainly are Chinese herfunctionall medicine.Function of claims is still the traditional Chinese medicine formulation, lack combination with modern Western medicine and the nutrition. For example, the function of antioxidant, the functional food ingredients seldom have antioxidant system evaluation, especially litter research about the relationship among the antioxidant functional food claims and the raw materials and components.As China’s most authoritative food composition datafunctionalse-China food composition is also lack of this part of the data. Therefore, the research about the raw materials often used in China functional food, especially its nutritional content,the functional components and the relationship between composition and antioxidant activity is under the circumstances.Objectives1. Add data to Chinese food composition table, fill data gaps of functional food nutritional ingredients, functional components.2. Using the in vitro antioxidant evaluation method to evaluate the antioxidant activity of common raw materials in functional food, while make aid evaluation for antioxidant activity evaluation to common raw materials about functional food by a-glucosidase inhibitory activity.3. Observe the relationship and effectiveness between the nutrients, functional factors and antioxidant activity, provide the scientific functionalsis for functional food claim. 4. Selecting the high antioxidant activity of functional food raw materials make the guidance for using of functional food raw materials in future5. Apply the statistical methods to construct regression model between functional components and the in vitro antioxidant activityMaterial and MethodsOn the functionalsis of the traditional Chinese medicine and Medicinal and Edible theories, according to the inform of "to further standardize the management of raw materials to functional food of Ministry of Functional "(Document No.51), we selected170species common raw materials including plants, animals, involving a total of65subjects in our study.30species are belong to Document No.51annex I,85species belonging to Annex Ⅱ, and the remaining55species belong to other common functional food raw materials, mainly are traditional Chinese herfunctionall medicine.1. Analysis of the functionalsic nutrientsAdopt national standards to analyze nutrition Determination of170kinds of common raw materials of functional food. Water used the method of GB/T5009.3-2003, protein is GB/T5009-2003)、ash is GB/T5009.4-2003, magnesium used GB/T5009.90-2003, potassium used GB/T5009.91-2003, sodium used GB/T5009.91-2003, phosphorous used GB/T5009.87-2003, iron used GB/T5009.90-2003, Calcium used Atomic absorption spectrophotometry(GB/T5009.92-2003), fat used Soxhlet extractor method(GB/T5009.6-2003), vitamin A used GB14750-1993, vitamin E used HPLC method(GB/T5009.82-2003), vitamin C used fluorometric method (GB/T5009.86-2003), thiamine used GB/T5009.84-2003, Riboflavin used fluorometric method (GB/T5009.85-2003)。2. Analysis of functional componentsTotal flavonoids determination is Aluminum chloride colorimetric assay, total saponins used Vanillin perchloric acid assay and total alkaloids used Acid dye colorimetry.According different plant medicinal part, classify the raw materials and discussed the distribution of nutrients, medicinal ingredients in different parts3. Antioxidant activity evaluation and high-throughput screening of a-glucosidase inhibited activityRespectively adopting the most advanced in vitro antioxidant evaluation method-ORAC method and the classical DPPH· radical scavenging method for the common functional fodd raw material. We reformed the DPPH· radical scavenging method by the automatic biochemical analyzer to improve experimental conditions to achieve a high throughput screening. DPPH· free radical removal efficiency is expressed by the concentration of IC50before, and now, after the mathematical conversion, the total antioxidant capacity (TAC value) is the express result,easy to understand and more clear and intuitive, using p-NPG method, p-NPG as substrate to screen a-glucosidase inhibit activity of170kinds of functional food raw materials, using automatic biochemical Analyzer to achieve high-throughput screening, a-glucosidase inhibition efficiency is expressed by50%inhibitory ration needed the raw materials concentration (IC50value). According to different medicinal parts, observe the antioxidant activity level in different medicinal parts. Screening the raw material with high Antioxidant capacity and high α-glucosidase inhibitory activity. According to traditional Chinese medicine theory, classify the common functional food raw materials in accordance with the four properties, five flavors and yin and yang, to observe the different categories of the level of antioxidant activity.4. Correlation of Nutritional composition, functional components and antioxidant activityCorrelation research is developed by SPSS software, spearman method for the nutritional content, functional and antioxidant activity components of170species of raw materials, results showed that antioxidant activity and flavonoids are most closely. Using SPSS software Classify Hierarchical Cluster modules, clustering principle indicator cluster (R type) in the correlation coefficient distance (Pearson correlation), methods for the class on average chain method (Between-groups linkage) clustering of all raw materials analysis, comparing the degree of similarity between different indicators. According to cluster analysis and correlation analysis results, further use of SPSS software, the Regression module, to establish the regression equation, flavonoids as independent variables and ORAC value as dependent variable.Research result1. Basic nutrition ingredients Determination of170kinds of common functional food raw materialsFollowing on Determination of170kinds of common functional food nutrients ingredients, we found that nutrients are abundant and widely distributed. Among Macronutrients, the water content distribution is in the0.5g-69.5g/100g, with a maximum water content of fresh Rhizoma Imperatae and royal jelly; Protein in the name of "benevolence","seed" category of raw material were higher, mostly distributed in fruits, seeds. Ash mainly exist in animal raw materials; iron is distributed in different parts of plant, To free oneself (cicada larvae into adults under the shell when the slough) was highest (1634.7mg/100g), is the lowest of the royal jelly of1256times. More magnesium present in the raw materials of animal and plant part of above ground, between the range of3.26-2636.7mg/100g. Potassium is mainly distributed in the ground part of the raw materials; Compositae Eclipta has the highest concentrations. Sodium mainly exist in animal samples, the highest concentrations is sea cucumber. Phosphorus is distributed in the roots to the highest content of Rhodiola and Siberian, and their analysis parts are the roots. The highest calcium content of raw materials is the Cuttlebone (dried squid squid related species within the shell), and in the plant raw material is more at leaf. Vegetable fats are generally essential fatty acids, linoleic acid and linolenic acid, after Soxhlet extraction method and we found that170kinds of raw materials contain a certain amount of fat, changes in range between0.09-53.7g/100g, more present in the plant fruit, seeds, such as perilla seed, hemp seed, and some plant material in the fat content even higher than the animal samples. After determination of vitamin analysis we found that subjects have generally low levels of vitamin, especially vitamin A, vitamin C and vitamin E, in most of the raw materials or even not detected, which may be due to raw materials are processed professionally in the sun, washing and bring loss.2. Determination results of functional ingredientsFlavonoids, saponins are the most frequent claim components in functional food. Therefore, in this study, they are chosen as the main target, and on this basis, select the alkaloids of this ubiquitous component of Chinese herbs while a total of three indicators170kinds of raw materials for the determination of functional components. After the analysis, the percentage of total flavonoids, total saponins and total alkaloids of170kinds of functional food showed:Most of the material in the total flavonoids, saponins were significantly different and varied greatly. The top10highest total flavonoid content of raw materials were FlosSophorae Sophoricoside, Magnolia, Acanthopanax, perilla, Chinese rose, Albizia julibrissin, Hong Ru, the leaves and myrobalan; The total saponin content of samples of the top10were Kudingcha, Anemarrhenae, Achyranthes bidentata, Suanzaoren, raspberries, American ginseng, Evodia, forsythia and Fructus Alpiniae Oxyphyllae. In Weight loss function food, total alkaloids are major components, through the determination of the levels found generally low, and little difference in content; top10samples were anti has, berberine, aloe, Hong Gapi, American ginseng, fruit of Chinese honeylocust, ephedra, cannabis Rob, and Bupleurum ChuanxiongAfter the analysis of the raw material in accordance with the medical site classification and then to observe the different parts of the content of functional components and we found:a high flavonoid content in plants generally flower parts, fruits and leaves, flowers, high content of total flavonoids in general, the average content of1.42%; saponins are secondary metabolites of plants to the leaves of more abundant, the average content was1.65%; roots and rhizomes of the class highest alkaloid content. Flavonoids, saponins and alkaloids in general does not exist in animal samples.3. Antioxidant activity and inhibition of a-glucosidase activity of raw materials in China Functional foodAntioxidant function is one of the important functions of functional food. Evaluation of antioxidant activity often choose two kinds of methods or more were determined simultaneously. Our study chose traditional DPPH· radical scavenging method and the current world’s most advanced ORAC method to analyze for170kinds of commonly used functional food raw materials. DPPH· radical is a stable chemical radicals, at the presence of antioxidants, antioxidants and their single-electron pair and its absorption gradually fade, and their acceptance of its fading into the quantitative relationship between the number of electrons, with spectrophotometric changes in absorbance calculated by free radical scavenging activity of samples, that is antioxidant activity. In our study, with the automatic biochemical analyzer, DPPH· method of traditional experimental conditions were improved to achieve a high throughput screening, and to avoid the free radical damage to human body. By measuring,23kinds of raw materials basically no free radical scavenging activity, and the remaining raw materials are some free radical scavenging activity, and there are16kinds of raw materials more active than the positive control of vitamin C, according to the order of their activity from high to low are epimedium, Cinnamomum Ramulus, Dendrobium, vine of multiflower knotweed, system Cistanche (system large Yun), Evodia, lotus leaf, Rob cannabis, forsythia, Terminalia, group cuttlebone, roses, Sophoricoside, wild buckwheat, Aloe vera and Centella asiatica, the weakest scavenging activity of all raw materials is the semen sojae praeparatum; through mathematics the IC50values of free radical scavenging activity converted to TAC value, which avoids the radical scavenging activity (IC50values) comparison difficult, but also eliminates the effect of raw material concentration on the expression of clear. ORAC assay is recommended by the international evaluation of new anti-oxidation. After ORAC analyze, the170kinds of raw materials, water-soluble ORAC value are between0.27～3579μmol TE/g. the highest ORAC value is Kudingcha, the lowest is the pearl, the other hith values samples are aloe, FlosSophorae Evodia, Sophoricoside, Danshen; the low values samples are cuttlebone, abalone, bird’s nest, sea cucumber (Gypsophila), Gorgon, Polyporus, Red Deer fetus etc. for the ORAC value of the minimum of raw materials, and more for the animal samples. We used SPSS software Correlation module to develop statistical correlation analysis, we found clear DPPH·free radical method the results obtained with the results measured by ORAC method has some relevance, that means these two kinds of method in the evaluation of antioxidant activity have the same trend. From the analysis of different parts, we found that free radical scavenging effect of better raw materials are mainly raw materials like flowers, with an average IC50value of0.99mg/mL, followed by leaf; ORAC value are classified according to their medicinal part, found that stem, leaves material although a small number, but with the highest ORAC average values. α-glucosidase is a very important enzymes, through the hydrolysis of a-1,4glycosidic bond from the non-reducing end of polysaccharide glucose cut. The body of the starch, dextrin, sucrose and other carbohydrates in the use of absorption depends on the intestinal brush border on the enzyme activity, control of diabetes and the need to reduce the activity of the enzyme, a-glucosidase inhibitors by competitive inhibition of a-glucosidase activity, can block two-sugar sugar solution into a single delay the absorption of sugar, so blood sugar steady and slow to maintain a certain level. In our study, acarbose is used as the positive control, using the classic method of p-NPG for170kinds of raw materials on the a-glucosidase inhibitory activity screening test, the use of automatic biochemical analyzer to achieve high-throughput screening and more efficient than96microplates. The results showed that a total of84raw materials has some inhibitory activity, of which25species of inhibitory activity was stronger than positive control acarbose. Overall, Chinese herbal medicine as an α-glucosidase inhibitor, the more complex compositions lead to mechanism needs further study.About inhibiting a-glucosidase activity, in the the top20raw material, there are9kinds of raw materials both strong suppression of α-glucosidase activity and the high ORAC value. They were roses, Terminalia, Fagopyrum, Epimedium, vine of multiflower knotweed, Apocynum leaves, cuspidatum, Chinese rose with Radix. There are8kinds of raw materials both strong suppression of a-glucosidase activity and strong TAC value. They are epimedium, vine of multiflower knotweed, Rob cannabis, Terminalia, roses, wild buckwheat, red Shao and Apocynum leaves.8both have a high ORAC value and a highTAC values were roses, Terminalia, wild buckwheat, Epimedium, vine of multiflower knotweed, Apocynum leaves, vines,vine of multiflower knotweed and aloe. In the future, these materials need more research and application to play important role.4.Antioxidant activity relationship with the theory of Traditional Chinese MedicineTCM theory holds that objects have "four properties" of the cold, hot, warm, cool and "five flavors " that is acrid, sweet, sour, bitter, salty, and also often use the "Yin","Yang" to evaluate the effects of Chinese herbs. In this study, the functional food are mainly herbal-based raw materials, so with four properties, five flavors and yin and yang were classified, we listed each type of material corresponding ORAC value. After research, we found:Cold material compared to the hot, warm and cool materials, the ORAC value of the highest, but the quantity is not much; bitterness of raw materials compare to the acrid, sweet, sour, salty class of its ORAC value not only high, quantity is greater; yin materials with high ORAC value and the number is greater. After further analysis of variance, we found that sour, bitter class materials ORAC value have significantly different (p<0.05), but by four properties classification, the raw materials of their ORAC values were no significant differences (p>0.05); yin yang kind of raw materials and class materials ORAC values significantly different (p<0.05), showed that the antioxidant activity by Yin Yang is more reliable classification.3.1The correlation analysis, cluster analysis regression equation SPSS software Correlation modules, Classify module and Regression modules respectively are used in nutrition, functional components and ORAC value, TAC values of the correlation analysis, cluster analysis and linear regression analysis. The results showed that:ORAC value and total flavonoids, total saponin content have some correlation, correlation coefficients were0.500、0.288(p<0.01), while the alkaloid content was almost no correlation with the ORAC value. TAC values and flavonoids, saponins also have some correlation coefficients were0.314、0.208(p <0.01), total flavonoids, total saponin and inhibited α-glucosidase activity of the IC50value (mg/mL) showed a negative correlation, shows that flavonoids, saponin content is more, the inhibition of α-glucosidase activity is stronger, and the IC50value is more lower, which is consistent with theory, though not statistically significant.Cluster analysis is based on characteristics of the thing itself, individual classification is based on the principle of feather flock together something categories. Used SPSS software Classify-Hierarchical cluster module cluster to analyze the different indicators and we found that flavonoids and ORAC values of the short distance in the statistical relationship between the tree, therefore, the flavonoid content can be regarded as the main ORAC value effect factors, other factors are in turn based on distance close to the level of saponin, TAC values and alkaloids; α-glucosidase inhibitory activity with IC50value of total alkaloids and a tree closest relationship, followed by the flavonoids and saponins.Using SPSS software Classify-Hierarchical cluster module to classify170kinds of raw materials based on antioxidant capacity index, then they are divided into six categories, which, because of ORAC value of the highest Folium llicis Latifoliae automatically classified as Class1, is defined as "very strong antioxidant "class; Flos Sophorae Immaturus and Aloe (ORAC mean2667μmol TE/g) of the two raw materials automatically classified as category2, defined as" the strong antioxidant capacity "level; to Salvia, Pueraria, represented by15kinds of raw materials (ORAC average1260μmol TE/g) classified as Category3, defined as "high antioxidant capacity"; to Chinese rose, Citrus aurantium, represented by25types of raw materials classified as Category4, ORAC mean723μmol TE/g, defined as "antioxidant capacity"; to senna, Saussurea costus represented47categories classified as Category5, ORAC mean314μmol TE/g, is defined as "weak antioxidant capacity";6category pearl, Radix Aucklandiae and other raw materials, a total of80species, ORAC mean40.5μmol TE/g, is defined as "a very weak antioxidant activity."ConclusionChina’s functional food raw material is widely used, evolved many families and distribution. Many kinds of medicinal parts of more nutritious ingredients, big difference in the effectiveness of ingredients, different parts of the nutrients, functional components were significant differences in content, but a distribution of components in the same position with convergence, such as moisture, protein, ash, mainly in the animal raw materials, flavonoid, saponins, alkaloids mainly present in plant materialAfter the traditional method of DPPH free radical scavenging screening, we found among170kinds of common functional food raw materials, there are152have scavenging activity and there are16kinds of raw materials higher than the positive control activity of vitamin C;170kinds of raw materials ORAC value is quite different, ORAC value material difference between the highest and lowest13255times. Free radical scavenging method and the ORAC method were significantly correlated, indicating that the same trend in two ways. There were selected from84kinds of α-glucosidase inhibitory activity of raw materials, including25species of inhibitory activity was stronger than positive control acarbose. Chinese herbal medicine are all major parts of α-glucosidase inhibition, Chinese herbal medicine as an α-glucosidase inhibitor mechanism of the onset is more complex, needs further study, α-glucosidase inhibition activity is higher in the top20raw materials, raw materials are9strong inhibition of both α-glucosidase activity was also highly ORAC value; are eight kinds of raw materials both strong a-glucosidase inhibitory activity, while TAC has a strong value.According to the Chinese "four properties","five flavors ","Yin Yang" Theory and Comparative antioxidant activity of the classified study found cold, cool, negative raw materials usually have high ORAC value. After analysis of variance showed that by " four properties " of classified material whose ORAC value is no significant difference (p>0.05), through the "five flavors" for the raw materials of its ORAC value groups were significantly different (p<0.05), that by " four properties "to distinguish between antioxidant activity is open to question;"yin "type of raw materials and" yang "type of raw materials ORAC values significantly different (p <0.05), showed that the antioxidant activity through the yin and yang is a more reliable classification.After statistical correlation analysis, the result showed that the total flavonoids, total saponin content and ORAC value of percentage of a certain correlation (p<0.01), in particular flavonoids and a high correlation coefficient; alkaloid content and ORAC value is the basic no correlation. Flavonoids, saponins and free radical scavenging activity has some correlation (p<0.01), total flavonoids, saponins and inhibition of a-glucosidase activity of the IC50value (mg/mL) showed a negative correlation, indicating that flavonoids, saponins with the higher content, the inhibitory activity is more stronger and the IC50value is more smaller; and alkaloid content was almost no correlation with the activity. After statistical clustering analysis showed that flavonoids and ORAC values in the statistical relationship between the short distance and have the tree, therefore, affect the flavonoid anti-ORAC value may be regarded as a major factor, other factors are in turn based on distance close to the level of saponin, TAC value, and alkaloids; α-glucosidase inhibitory activity with IC50value of alkaloids and a tree closest relationship, followed by the composition of flavonoids and saponins.170kinds of raw materials based on antioxidant capacity index, using SPSS soft ware, then they are divided into six categories, which, because of ORAC value of the highest Folium llicis Latifoliae automatically classified as Class1, is defined as "very strong antioxidant "class; Flos Sophorae Immaturus and Aloe (ORAC mean2667μmol TE/g) of the two raw materials automatically classified as category2, defined as" the strong antioxidant capacity "level; to Salvia, Pueraria, represented by15kinds of raw materials (ORAC average1260μmol TE/g) classified as Category3, defined as "high antioxidant capacity"; to Chinese rose, Citrus aurantium, represented by25types of raw materials classified as Category4, ORAC mean723μmol TE/g, defined as "antioxidant capacity"; to senna, Saussurea costus represented47categories classified as Category5, ORAC mean314μmol TE/g, is defined as "weak antioxidant capacity";6category pearl, Radix Aucklandiae and other raw materials, a total of80species, ORAC mean40.5μmol TE/g g, is defined as "a very weak antioxidant activity."InnovationsFirst time, we analyzed the functional food raw materials about their nutrients and functional components at large extent for the Chinese food composition and meanwhile, we fill the gaps in this part of the data, providing the basic data for future use.For the first time by multi-functional microplate reader and automatic biochemical analyzer respectively, used ORAC test and high-throughput DPPH· radical scavenging for herbs by applying complex evaluation of antioxidant activity of raw materials, the first time the use of automatic biochemical analyzer to the a-glucosidase inhibitors in high throughput screening.The first time to compare high antioxidation ability and high a-glucosidase inhibitory activity of samples.The first time, applied the cluster analysis and evaluation model for in vitro antioxidant acitivity and composition, providing reference to applications functional food raw materials in future.