COMPOUND COMPOSITION WITH ANTI-OXIDATION AND ANTI-AGING EFFECTS AND PREPARATION METHOD AND APPLICATION THEREOF

Abstract

A compound composition with anti-oxidation and anti-aging effects and preparation method and application thereof we disclosed. 5-8 kinds of traditional Chinese medicine extracts from separate extraction of Panax ginseng C. A. Meyer, Poria, Rehmannia glutinosa, Cistanche deserticola Ma, Salviae Miltiorrhizae, Codonopsis Radix, Persicae Semen and Ophiopogon japonicus (Linn.f.) Ker-Gawl. are compounded with chitooligosaccharide to prepare the compound composition with anti-oxidation and anti-aging effects. The compound composition has no toxic side effects, is easy to be absorbed, and can be used for delaying aging, repairing DNA damage and anti-oxidation damage, and can also reduce the risk of geriatric diseases. Moreover, the compound composition can be used as a raw material of dietary supplement or health food to improve telomerase content and delay aging. Therefore, the compound composition is suitable for promotion and application.

Claims

1. A compound composition with anti-oxidation and anti-aging effects comprised of traditional Chinese medicine extracts and chitooligosaccharides according to a mass ratio of (3-8): 1; wherein the traditional Chinese medicine extracts are prepared by extracting the following Chinese medicinal materials respectively and compounding, according to a mass percentage: 7%-15% of Panax ginseng C. A. Meyer; 14%˜35% of Poria; 14%˜35% of Rehmanniae glutinosa; 14%˜35% of Cistanche deserticola Ma; 7%˜15% of Salviae Miltiorrhizae; 0%˜10% of Codonopsis Radix, 0%˜10% of Persicae Semen; and 0%˜10% of Ophiopogon japonicus (Linn.f.) Ker-Gawl.

2. The compound composition of claim 1, wherein the traditional Chinese medicine extracts are prepared by extracting the following Chinese medicinal materials respectively and compounding, according to a mass percentage: 12%-15% of Panax ginseng C. A. Meyer, 25%˜30% of Poria; 20%˜25% of Rehmanniae glutinosa; 20%˜25% of Cistanche deserticola Ma; 12%˜15% of Salviae Miltiorrhizae; 2%˜3% of Codonopsis Radix, 1%˜2% of Persicae Semen; and 1%˜2% of Ophiopogon japonicus (Linn.f.) Ker-Gawl.

3. The compound composition of claim 1, wherein the traditional Chinese medicine extracts ae prepared by extracting the following Chinese medicinal materials respectively and compounding, according to a mass percentage: 13% of Panax ginseng C. A. Meyer; 26% of Poria; 22% of Rehmanniae glutinosa; 22% of Cistanche deserticola Ma; 13% of Salviae Miltiorrhizae; 2% of Codonopsis Radix; 1% of Persicae Semen; and 1% of Ophiopogon japonicus (Linn.f.) Ker-Gawl.

4. A method of preparing the compound composition of claim 1, comprising: subjecting Panax ginseng C. A. Myer, Poria, Rehmanniae glutinosa, Cistanche deserticola Ma, Salviae Miltiorrhizae, Codonopsis Radix, Persicae Semen and Ophiopogon japonicus (Linn.f.) Ker-Gawl, to extraction, purification, concentration and drying to produce Panax ginseng C. A. Meyer extract, Poria extract, Rehmanniae glutinosa extract, Cistanche deserticola Ma extract, Salviae Miltiorrhizae extract, Codonopsis Radix extract, Persicae Semen extract and Ophiopogon japonicus (Linn.f.) Ker-Gawl, extract; and mixing the Panax ginseng C. A. Myer extract, the Poria extract, the Rehmanniae glutinosa extract, the Cistanche deserticola Ma extract, the Salviae Miltiorrhizae extract, the Codonopsis Radix extract, the Persicae Semen extract, the Ophiopogon japonicus (Linn.f.) Ker-Gawl, extract and the chitooligosaccharides uniformly to produce the compound composition; wherein the method specifically comprises: (1) crushing, sieving, and subjecting Panax ginseng C. A. Meyer, Poria, Rehmanniae glutinosa, Cistanche deserticola Ma, Salviae Miltiorrhizae, Codonopsis Radix, Persicae Semen and Ophiopogon japonicus (Linn.f.) Ker-Gawl, to extraction respectively with an ethanol solution under refluxing and filtration; subjecting a residue to extraction with a solvent according to certain proportions and filtration several times; and combining filtrates to obtain a crude Panax ginseng C. A. Meyer extract, a crude Poria extract, a crude Rehmanniae glutinosa extract, a crude Cistanche deserticola Ma extract, a crude Salviae Miltiorrhizae extract, a crude Codonopsis Radix extract, a crude Persicae Semen extract and a crude Ophiopogon japonicus (Linn.f.) Ker-Gawl, extract, respectively; (2) subjecting the crude Panax ginseng C. A. Meyer extract, the crude Poria extract, the crude Rehmanniae glutinosa extract, the crude Cistanche deserticola Ma extract, the crude Salviae Miltiorrhizae extract, the crude Codonopsis Radix extract, the crude Persicae Semen extract and the crude Ophiopogon japonicus (Linn.f.) Ker-Gawl, extract to purification and concentration, spray drying or vacuum drying, and sieving to produce the Panax ginseng C. A. Meyer extract, the Poria extract, the Rehmanniae glutinosa extract, the Cistanche deserticola Ma extract, the Salviae Miltiorrhizae extract, the Codonopsis Radix extract, the Persicae Semen extract, the Ophiopogon japonicus (Linn.f.) Ker-Gawl extract; and (3) mixing the Panax ginseng C. A. Meyer extract, the Poria extract, the Rehmanniae glutinosa extract, the Cistanche deserticola Ma extract, the Salviae Miltiorrhizae extract, the Codonopsis Radix extract, the Persicae Semen extract, the Ophiopogon japonicus (Linn.f.) Ker-Gawl, extract and the chitooligosaccharides uniformly in the weight ratio of claim 1 to produce the compound composition with anti-oxidation and anti-aging effects.

5. The method of preparing the compound composition of claim 4, wherein in the step (1), the sieving is performed using a sieve of 20-80 mesh; a volume ratio of Panax ginseng C. A. Meyer to the ethanol solution is 1:(5˜20); a volume ratio of Poria to the ethanol solution is 1:(5˜20); a volume ratio of Rehmanniae glutinosa to the ethanol solution is 1:(5˜20) a volume ratio of Cistanche deserticola Ma to the ethanol solution is 1:(5˜20); a volume ratio of Salviae Miltiorrhizae to the ethanol solution is 1:(5˜20); a volume ratio of Codonopsis Radix to the ethanol solution is 1:(5˜20); a volume ratio of Persicae Semen to the ethanol solution is 1:(5˜20); a volume ratio of Ophiopogon japonicus (Linn.f.) Ker-Gawl to the ethanol solution is 1:(5˜20); and the extraction of Panax ginseng C. A. Meyer, Poria, Rehmanniae glutinosa, Cistanche deserticola Ma, Salviae Miltiorrhizae, Codonopsis Radix, Persicae Semen and Ophiopogon japonicus (Linn.f.) Ker-Gawl, is performed 1-3 times at 80-100° C. for 1-3 h each time, respectively.

6. The method of preparing the compound composition of claim 4, wherein in the step (2), the spray drying is performed under 180-200° C. the vacuum drying is performed under 50-100° C.; and the sieving is performed using a sieve of 60-80 mesh.

7. The method of preparing the compound composition of claim 6, wherein the step (2) further comprises: before the spray drying or the vacuum drying, introducing β-cyclodextrinto the concentrated Panax ginseng C. A. Meyer filtrate, Poria filtrate, Rehmanniae glutinosa filtrate. Cistanche deserticola Ma filtrate, Salviae Miltiorrhizae filtrate, Codonopsis Radix filtrate, Persicae Semen filtrate and Ophiopogon japonicus (Linn.f.) Ker-Gawl, filtrate, respectively, followed by mixing uniformly; wherein the β-cyclodextrinto is 1%-20V by weight of Panax ginseng C. A. Meyer, Poria, Rehmanniae glutinosa, Cistanche deserticola Ma, Salviae Miltiorrhizae, Codonopsis Radix, Persicae Semen and Ophiopogon japonicus (Linn.f.) Ker-Gawl, respectively.

8. An application of the compound composition of claim 1 in a preparation of health food.

9. The application of claim 8, wherein the compound composition is used as a raw material of dietary supplement or health food, and a dosage form of the compound composition is a capsule, a granule or a tablet.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0074] In order to explain the embodiments of the present disclosure or the technical solutions in the prior art more clearly, the following drawings that need to be used in the description of the embodiments or the prior art will be briefly introduced. Obviously, the drawings in the following description ae only embodiments of the present disclosure. For those of ordinary skill in the art, other drawings can be obtained based on the drawings disclosed without creative work.

[0075] FIG. 1 is the scavenging ability on DPPH free radical of the compound composition of the disclosure.

[0076] FIG. 2 is the scavenging ability on hydroxyl radical of the compound composition of the disclosure.

DETAILED DESCRIPTION OF EMBODIMENTS

[0077] Technical solutions of the present disclosure will be clearly and completely described below with reference to the embodiments. Obviously, described below we merely some embodiments of the disclosure, which are not intended to limit the disclosure. Other embodiments made by those skilled in the art without sparing any creative effort should fall within the scope of the disclosure.

The embodiments of the disclosure provide a compound composition, which has no toxic side effect and is easy to be absorbed. The compound composition is used as a raw material of dietary supplement or health food for delaying aging and anti-oxidation. In addition, the compound composition can increase the telomerase content, repair DNA damage and anti-oxidation damage, and can also reduce the occurrence risk of geriatric diseases. The compound composition is suitable for market promotion.

[0078] The disclosure will be further described below with reference to the embodiments. It should be understood that these embodiments are merely illustrative of the disclosure, and are not intended to limit the disclosure. Any improvement and modification made by those skilled in the art without departing from the spirit of the disclosure should still fall within the scope of the disclosure.

Hereinafter, the technical solution disclosed will be further described with reference to specific embodiments.

Embodiment 1

[0079] A compound composition with anti-oxidation and anti-aging effects is formulated as follows:

[0080] chitooligosaccharides: 50 g, Panax ginseng C. A. Meyer medicinal material: 100 g, Poria medicinal material: 200 g, Rehmannia glutinosa medicinal material: 166 g, Cistanche deserticola Ma medicinal material: 168 g, Salviae Miltiorrhizae medicinal material: 100 g. magnesium stearate: 5 g.

[0081] The specific preparation steps of the compound composition are as follows:

[0082] (1) The Panax ginseng C. A. Meyer medicinal material is crushed into coarse granules (sieved with a sieve of 20 mesh). The coarse granules were subjected to extraction with a 75% ethanol solution in a volume ratio of 1:8 under refluxing twice for 3 h each time and filtration. Filtrates were combined, purified, concentrated and added with 10% β-cyclodextrinto. The combined filtrate was continuously subjected to spray drying at 200° C. The dried product was sieved with a sieve of 80 mesh to produce the Panax ginseng C. A. Meyer extract.

[0083] (2) The Poria medicinal material is crushed into coarse granules (sieved with a sieve of 80 mesh). The coarse granules were subjected to extraction with water in a volume ratio of 1:10 under refluxing once for 100 min and filtration. Filtrate was concentrated to 1/10 of the original volume, precipitated with 80% ethanol solution for 12 h and filtrated. Then filtrate was added with 10% β-cyclodextrinto and continuously subjected to spray drying at 200° C. The dried product was sieved with a sieve of 80 mesh to produce the Poria extract.

[0084] (3) The Rehmanniae glutinosa medicinal material is crushed into coarse granules (sieved with a sieve of 20 mesh). The coarse granules were subjected to extraction with a 60% ethanol solution in a volume ratio of 1:8 under refluxing twice for 1.5 h each time and filtration. Filtrates were combined, purified, concentrated and added with 10% β-cyclodextrinto. The combined filtrate was continuously subjected to spray drying at 200° C. The dried product was sieved with a sieve of 80 mesh to produce the Rehmanniae glutinosa extract.

[0085] (4) The Cistanche deserticola Ma medicinal material is crushed into coarse granules (sieved with a sieve of 20 mesh). The coarse granules were subjected to extraction with a 75% ethanol solution in a volume ratio of 1:8 under refluxing twice for 1 h each time and filtration. Filtrates were combined, purified, concentrated and added with 10% β-cyclodextrinto. The combined filtrate was continuously subjected to spray drying at 200° C. The dried product was sieved with a sieve of 80 mesh to produce the Cistanche deserticola Ma extract.

[0086] (5) The Salviae Miltiorrhizae medicinal material is subjected to extraction with an 80% ethanol solution in a volume ratio of 1:8 under refluxing twice for 1.5 h each time and filtration. Filtrates were combined, purified, concentrated and added with 10% β-cyclodextrinto. The combined filtrate was continuously subjected to spray drying at 200° C. The dried product was sieved with a sieve of 80 mesh to produce the Salviae Miltiorrhizae extract.

[0087] (6) The prepared traditional Chinese medicine extract and chitooligosaccharide were evenly mixed, granulated and filled into compound capsules, which is 100 times of the daily prescription dose.

Embodiment 2

[0088] A compound composition with anti-oxidation and anti-aging effects is formulated as follows:

[0089] chitooligosaccharides: 40 g, Panax ginseng C. A. Meyer medicinal material: 72 g. Poria medicinal material: 144 g, Rehmannia glutinosa medicinal material: 120 g, Cistanche deserticola Ma medicinal material: 180 g. Salviae Miltiorrhizae medicinal material: 72 g. magnesium stearate: 3 g.

[0090] The specific preparation steps of the compound composition are as follows:

[0091] (1) The Panax ginseng C. A. Meyer medicinal material is crushed into coarse granules (sieved with a sieve of 20 mesh). The coarse granules were subjected to extraction with a 75% ethanol solution in a volume ratio of 1:6 under refluxing twice for 3 h each time and filtration. Filtrates were combined, purified, concentrated and added with 10% β-cyclodextrinto. The combined filtrate was continuously subjected to spray drying at 200° C. The dried product was sieved with a sieve of 80 mesh to produce the Panax ginseng C. A. Meyer extract.

[0092] (2) The Poria medicinal material is crushed into coarse granules (sieved with a sieve of 80 mesh). The coarse granules were subjected to extraction with water in a volume ratio of 1:8 under refluxing once for 120 min and filtration. Filtrate was concentrated to 1/10 of the original volume, precipitated with 80% ethanol solution for 12 h and filtrated. Then filtrate was added with 10% β-cyclodextrinto and continuously subjected to spray drying at 200° C. The dried product was sieved with a sieve of 80 mesh to produce the Poria extract.

[0093] (3) The Rehmanniae glutinosa medicinal material is crushed into coarse granules (sieved with a sieve of 20 mesh). The coarse granules were subjected to extraction with a 70% ethanol solution in a volume ratio of 1:10 under refluxing twice for 1 h each time and filtration. Filtrates were combined, purified, concentrated and added with 10% β-cyclodextrinto. The combined filtrate was continuously subjected to spray drying at 200° C. The dried product was sieved with a sieve of 80 mesh to produce the Rehmanniae glutinosa extract.

[0094] (4) The Cistanche deserticola Ma medicinal material is crushed into coarse granules (sieved with a sieve of 20 mesh). The coarse granules were subjected to extraction with a 80% ethanol solution in a volume ratio of 1:6 under refluxing twice for 1 h each time and filtration. Filtrates were combined, purified, concentrated and added with 10% β-cyclodextrinto. The combined filtrate was continuously subjected to spray drying at 200° C. The dried product was sieved with a sieve of 80 mesh to produce the Cistanche deserticola Ma extract.

[0095] (5) The Salviae Miltiorrhizae medicinal material is subjected to extraction with an 80% ethanol solution in a volume ratio of 1:8 under refluxing twice for 1.5 h each time and filtration. Filtrates were combined, purified, concentrated and added with 10% β-cyclodextrinto. The combined filtrate was continuously subjected to spray drying at 200° C. The dried product was sieved with a sieve of 80 mesh to produce the Salviae Miltiorrhizae extract.

[0096] (6) The prepared extract and chitooligosaccharide are evenly mixed to obtain the traditional Chinese medicine compound composition, and the granules of the traditional Chinos medicine compound composition are obtained by mixing and granulating, which is 100 times of the daily prescription dose.

Embodiment 3

[0097] A compound composition with anti-oxidation and anti-aging effects is formulated as follows:

[0098] chitooligosaccharides: 30 g, Panax ginseng C. A. Meyer medicinal material: 63 g, Poria medicinal material: 103 g, Rehmannia glutinosa medicinal material: 84 g, Cistanche deserticola Ma medicinal material: 157 g, Salviae Miltiorrhizae medicinal material: 63 g. Codonopsis Radix: 52.5 g, Persicae Semen: 52.5 g, magnesium stearate: 5 g.

[0099] The specific preparation steps of the compound composition are as follows:

[0100] (1) The Panax ginseng C. A. Meyer medicinal material is crushed into coarse granules (sieved with a sieve of 20 mesh). The come granules were subjected to extraction with a 80% ethanol solution in a volume ratio of 1:4 under refluxing twice for 3 h each time and filtration. Filtrates were combined, purified, concentrated and added with 10% β-cyclodextrinto. The combined filtrate was continuously subjected to spray drying at 200° C. The dried product was sieved with a sieve of 80 mesh to produce the Panax ginseng C. A. Meyer extract.

[0101] (2) The Poria medicinal material is crushed into coarse granules (sieved with a sieve of 80 mesh). The coarse granules were subjected to extraction with water in a volume ratio of 1:6 under refluxing once for 1.5 h and filtration. Filtrate was concentrated to 1/10 of the original volume, precipitated with 80% ethanol solution for 12 h and filtrated. Then filtrate was added with 10% β-cyclodextrinto and continuously subjected to spray drying at 200° C. The dried product was sieved with a sieve of 80 mesh to produce the Poria extract.

[0102] (3) The Rehmanniae glutinosa medicinal material is crushed into coarse granules (sieved with a sieve of 20 mesh). The coarse granules were subjected to extraction with a 70% ethanol solution in a volume ratio of 1:6 under refluxing twice for 1.5 h each time and filtration. Filtrates were combined, purified, concentrated and added with 10% β-cyclodextrinto. The combined filtrate was continuously subjected to spray drying at 200° C. The dried product was sieved with a sieve of 80 mesh to produce the Rehmanniae glutinosa extract.

[0103] (4) The Cistanche deserticola Ma medicinal material is crushed into coarse granules (sieved with a sieve of 20 mesh). The coarse granules were subjected to extraction with an 80% ethanol solution in a volume ratio of 1:6 under refluxing twice for 1 h each time and filtration. Filtrates were combined, purified, concentrated and added with 10% β-cyclodextrinto. The combined filtrate was continuously subjected to spray drying at 200° C. The dried product was sieved with a sieve of 80 mesh to produce the Cistanche deserticola Ma extract.

[0104] (5) The Salviae Miltiorrhizae medicinal material is subjected to extraction with an 80% ethanol solution in a volume ratio of 1:8 under refluxing twice for 1.5 h each time and filtration. Filtrates were combined, purified, concentrated and added with 10% O-cyclodextrinto. The combined filtrate was continuously subjected to spray drying at 200° C. The dried product was sieved with a sieve of 80 mesh to produce the Salviae Miltiorrhizae extract.

[0105] (6) The Codonopsis Radix medicinal material is crushed into coarse granules (sieved with a sieve of 20 mesh). The coarse granules were subjected to extraction with a 60% ethanol solution in a volume ratio of 1:8 under refluxing once for 40 min and filtration. Filtrates were purified, concentrated and added with 10% β-cyclodextrinto. The combined filtrate was continuously subjected to spray drying at 200° C. The dried product was sieved with a sieve of 80 mesh to produce the Codonopsis Radix extract.

[0106] (7) The Persicae Semen is subjected to extraction with water in a volume ratio of 1:10 under refluxing once for 2 h. The extracted product were purified, concentrated and added with 10% β-cyclodextrinto. The combined product was continuously subjected to spray drying at 200° C. The dried product was sieved with a sieve of 80 mesh to produce the Persicae Semen extract.

[0107] (8) The prepared extract and chitooligosaccharide are evenly mixed to obtain the traditional Chinese medicine compound composition, and the granules of the traditional Chinese medicine compound composition are obtained by dry granulation, which is 100 times of the daily prescription dose.

[0108] Embodiments provided herein are described in a progressive manner, and each embodiment mainly focuses on the differences from other embodiments. The same or similar parts among various embodiments can be referred to each other.

[0109] Further, the following tests were performed to evaluate the efficacy of the compound composition prepared herein.

(1) Study on Anti-Oxidation Effect of the Disclosure In Vitro

[0110] The compound composition preparation and other drug prescriptions prepared in embodiment 1 were used to test the anti-oxidation effect of the compound composition in vitro.

[0111] 1) Grouping of Test Samples:

[0112] Test substance A: 50 g of chitooligosaccharides, 100 g of Panax ginseng C. A. Meyer medicinal material, 200 g of Poria medicinal material, 166 g of Rehmannia glutinosa medicinal material, 168 g of Cistanche deserticola Ma medicinal material, 100 g of Salviae Miltiorrhizae medicinal material, and 5 g of magnesium stearate were prepared into 100 times daily prescription dose according to the method of embodiment 1.

[0113] Test substance B: 50 g of chitooligosaccharides, 100 g of Panax ginseng C. A. Meyer medicinal material, 200 g of Poria medicinal material, 166 g of Rehmannia glutinosa medicinal material, 168 g of Cistanche deserticola Ma medicinal material, 33.3 g of β-cyclodextrinto, and 5 g of magnesium stearate were prepared into 100 times daily prescription dose according to the method of embodiment 1.

[0114] Test substance C: 50 g of chitooligosaccharides, 100 g of Panax ginseng C. A. Meyer medicinal material, 200 g of Poria medicinal material, 166 g of Rehmannia glutinosa medicinal material, 168 g of Codonopsis Radix, 60 g of Persicae Semen, and 5 g of magnesium stearate were prepared into 100 times daily prescription dose according to the method of embodiment 1.

[0115] Test substance D: 50 g of chitooligosaccharides, 100 g of Panax ginseng C. A. Meyer medicinal material, 200 g of Poria medicinal material, 166 g of Rehmannia glutinosa medicinal material, 100 g of Salviae Miltiorrhizae, 168 g of Codonopsis Radix, and 5 g of magnesium stearate were prepared into 100 times daily prescription dose according to the method of embodiment 1.

[0116] Test substance E: 50 g of chitooligosaccharides, 100 g of Panax ginseng C. A. Meyer medicinal material, 200 g of Poria medicinal material, 166 g of Rehmannia glutinosa medicinal material, 89.3 g of β-cyclodextrinto, 5 g of magnesium stearate were prepared into 100 times daily prescription dose according to the method of embodiment 1.

[0117] Preparation of test samples: the above test substances A-E were diluted with distilled water to four step concentrations of 10, 30, 50 and 100 mg/ml respectively.

[0118] 2) Test Method

[0119] a) DPPH Radical Scavenging Ability

[0120] 1 ml of sample solution and 1 ml of DPPH-ethanol solution (0.125 mol/L) were mixed in a vortex oscillator. After 30 min of dark reaction at room temperature, the absorbance value was measured at 517 nm. Anhydrous ethanol was used as the blank control, which was recorded as A. All samples were determined for 3 times in parallel, and the average value was taken. DPPH radical scavenging rate was calculated by the following formula:

[00001]$\mathrm{DPPH}\mathrm{radical}\mathrm{scavenging}\mathrm{rate}\left(\%\right)=\left(1-\frac{A_2-A_1}{A_0}\right)\times 100\%$

[0121] Where: A.sub.1 was the absorbance value of the sample without DPPH solution; A.sub.2 was the absorbance value of DPPH solution and sample.

[0122] b) Determination of Hydroxyl Radical Scavenging Rate

[0123] 1 ml sample solution was added with 1.0 ml FeSO.sub.4 (9 mmol/L), 10 ml salicylic acid ethanol solution (9 mmol/IL) and 1 ml H.sub.2O.sub.2 (9 mmol/L) respectively. After reaction at 37° C. for 1 h, the absorbance of the mixture at 510 nm was determined. Anhydrous ethanol was used as blank control, which was recorded as A.sub.0. All samples were determined for 3 times in parallel. The scavenging ability of hydroxyl radical was calculated by the following formula:

[00002]$\mathrm{Hydroxyl}\mathrm{radical}\mathrm{scavenging}\mathrm{rate}\left(\%\right)=\left(1-\frac{A_2-A_1}{A_0}\right)\times 100\%$

[0124] Where: A.sub.1 was the absorbance value of distilled water instead of H.sub.2O.sub.2; A.sub.2 was the absorbance value of hydroxyl radical system and sample.

[0125] 3) Result Analysis

[0126] DPPH is a kind of stable free radical with nitrogen as the center. There is a strong absorption peak at 517 nm. The electron or hydrogen atom provided by antioxidant reacts with DPPH, resulting in the decrease of its absorbance value at 517 nm, be change of absorbance value reflects the anti-oxidation ability of antioxidant.

[0127] It can be seen from FIG. 1 that the scavenging ability of DPPH radical of each test substance increased with the increase of concentration, and the anti-oxidation ability was as follows: test substance A>test substanceD>test substanceC>test substanceB>test substance E. Therefore, the anti-oxidation effect of test substance A was the best.

[0128] Hydroxyl radical is the most active chemical substance in the reactive oxygen system. It can easily pass through the cell membrane and react with biological molecules such as carbohydrate, protein, lipid, nucleic acid and almost any biological component in the cell to induce tissue damage or lead to cell death.

[0129] It can be seen from FIG. 2 that with the increase of the mass concentration of test substance, the scavenging ability of each test substance to hydroxyl radical presented a stable growth. The scavenging ability of test substance A was greater than that of other test substances in each mass concentration measurement range. When the mas concentration was 100.0 mg/ml, the scavenging ability of each test substance to hydroxyl radical reached the maximum.

[0130] Based on the above results, the disclosure has remarkable anti-oxidation effect. The indexes of test substance A were significantly better than those of the others, that is, the anti-oxidation effect of the compound composition composed of chitooligosaccharide. Panax ginseng C. A. Meyer, Poria, Rehmannia glutinosa, Cistanche deserticola Ma and Salviae Miltiorrhizae was significantly better than that of the compound composition composed of chitooligosaccharide, Panax ginseng C. A. Meyer, Poria, Rehmannia glutinosa and 0-1 kinds or two combinations of Cistanche deserticola Ma, Salviae Miltiorrhizae, Codonopsis Radix, Persicae Semen and Ophiopogon Japonicus (Linn.f.) Ker-Gawl. It shows that the anti-oxidation effect of the disclosure is related to the composition of the specific raw materials of the disclosure. The reasonable proportion of the raw materials of the disclosure, together with the specific raw material extraction method of the disclosure, makes the final prepared compound composition have significant anti-oxidation effect.

(2) Human Trial

[0131] The safety and anti-aging effect of the compound composition capsule prepared in embodiment 1 were tested.

[0132] Inclusion criteria: 50-75 years old, normal aging or disease-induced weakness, physical strength and lack of energy;

[0133] Exclusion criteria: persons with the following diseases: chronic allergic enteritis or functional diarrhea; autoimmune diseases or immunodeficiency; positive diagnosis of HIV, hepatitis B or hepatitis C; history of severe heart, lung, kidney and liver dysfunction; occurrence of major cardiovascular diseases in the past 6 months; neuropsychological diseases or cognitive impairment diagnosed by medicine; alcoholism and high alcohol intake.

[0134] Experimental design and grouping: before and after self-control.

[0135] Dosage and method: 3 capsules, 2 times a day, 3.6 g/day for 90 days.

[0136] Observation Indicators:

1) Security Observation:

[0137] {circle around (1)} Blood routine: red blood cell count, average red blood cell volume, red blood cell distribution width, average hemoglobin content, average red blood cell hemoglobin concentration, white blood cell count, monocyte count, monocyte ratio, neutrophil count, lymphocyte ratio, average platelet volume, hematocrit;

[0138] {circle around (2)} Urine routine: urine color, urine transparency, urine pH, red blood cells, white blood cells, urine protein, urine ketone body, urine sugar;

[0139] {circle around (3)} Blood biochemistry: liver function examination: total bilirubin, total protein, white protein, globulin, albumin/globulin ratio, aspartate aminotransferase, alanine aminotransferase, etc;

[0140] {circle around (4)} Renal function: creatinine (CR), blood urea nitrogen (BUN);

[0141] {circle around (5)} ECO.

2) Efficacy Observation:

[0142] {circle around (1)} Blood pressure skin test: skin elasticity, collagen and melanin abnormalities

[0143] {circle around (2)} Vascular regulatory factors: nitric oxide (No), vascular endothelial growth factor (VEGF)

[0144] {circle around (3)} Superoxide dismutase (SOD) activity and telomerase (TE) content in serum

[0145] {circle around (4)} Follow up service scale for experiential personnel: [0146] Aging symptoms: Four symptoms of poor physical strength (fatigue after walking or activities, poor physical strength in daily life, dizziness, poor sleep) were included, and were classified into grade I-IV according to the severity, with statistical integral values. [0147] Grade I (0 point): keeping a certain amount of exercise, energetic; daily energetic; rarely dizzy, only after overwork; falling asleep within 30 minutes, sleep time more than 5 hours, not easy to wake up, not sleepy during the day. [0148] Grade II (1 point): slightly tired after more walking activities (such as walking 2 km); slightly tired, not affecting daily activities; dizziness will occur only once or twice a month, life will not be affected; falling asleep within 30 min. sleep time is less than 5 h, no sleepiness during the day. [0149] Grade III (2 points): feeling between grade II and grade IV; fatigue easily, daily activities affected; dizziness twice a week, aggravating after fatigue, affecting daily life; falling asleep more than 30 minutes, sleeping less than 5 hours, easily awakened, sleepy at a fixed time during the day. [0150] Grade IV (3 points): weak back, dizziness, limb weakness; severe fatigue, severely restricted daily activities; dizziness every day, aggravated after a little activity, seriously affecting daily life; more than 30 minutes to sleep, sleep time less than 5 hours, often wake up in the middle of the night without reason, often sleepy during the day. [0151] Diseases of the elderly and sub-health symptoms: Six items (anemia, anorexia, cold infection, etc., mental unrest/palpitation, chills and pain) were included, and were classified into Grade I-IV according to the severity, with statistical integral values. [0152] Grade I (0 point): No symptoms. [0153] Level II (1 point): Low appetite for food, but normal eating; slight chills; slight anemia, which does not affect life after nutritional supplement, slight pain; only one or two times a month of mental unrest/palpitations; competent for general daily activities, in a little heavy physical activity, heart palpitations aggravated; occasionally cold, easy to fall ill only after season or tired. [0154] Grade III (2 points): difficulty in eating, risk of malnutrition; severe chills, significantly different from normal people; moderate anemia; severe pain in specific parts; restlessness/palpitation twice a week, slightly high activity (walking 2 km), severe palpitation, accompanied by mental distress; cold frequency between U and IV. [0155] Grade IV (3 points): no eating at all, severe malnutrition: extreme fear of cold, serious impact on life; severe anemia, maintaining normal life by drugs; general pain, serious impact on the quality of life; palpitation every day, mental tension, palpitation after slight activity; frequent cold, frequent illness, poor resistance.

TABLE-US-00001 TABLE 1 General condition before feeding Project Subjects Number 30 M/F 17/13 Age (years) 50~76

TABLE-US-00002 TABLE 2 Analysis of blood routine examination results before and after the test (x ± SD) Project 0 day 90 days P value WBC count (10.sup.9/L)   5.96 ± 3.00   6.91 ± 3.27 0.4729 Monocyte ratio (%)   9.06 ± 0.87   8.13 ± 1.47 0.3176 Monocyte count (10.sup.9/L)   0.66 ± 0.05   0.64 ± 0.17 0.7277 Lymphocyte ratio (%)  40.64 ± 12.07  40.55 ± 10.82 0.9861 WBC count (10.sup.9/L)   5.96 ± 3.00   6.91 ± 3.27 0.4729 Monocyte ratio (%)   9.06 ± 0.87   8.43 ± 1.47 0.3176 Monocyte count (10.sup.9/L)   0.66 ± 0.05   0.64 ± 0.17 0.7277 Lymphocyte ratio (%)  40.64 ± 12.07  40.55 ± 10.82 0.9861 Neutrophil ratio ratio (%)  57.99 ± 11.29  57.20 ± 13.98 0.8960 Neutrophil count (10.sup.9/L)   2.01 ± 1.33   2.20 ± 1.39 0.8123 Red blood cell distribution  11.06 ± 1.31  12.28 ± 1.41 0.1141 width (%) Mean corpuscular volume (fL)  90.98 ± 7.99  91.57 ± 8.38 0.8335 Mean platelet volume (fL)   9.97 ± 2.19  10.38 ± 1.66 0.6770 Hematocrit (%)   0.30 ± 0.07   0.31 ± 0.08 0.8535 Mean hemoglobin (pg)  31.95 ± 1.61  31.93 ± 1.82 0.9387 Mean hemoglobin 359.81 ± 1.63 353.81 ± 6.93 0.2187 concentration (g/L)

TABLE-US-00003 TABLE 3 Analysis of liver and kidney function and blood biochemical indexes before and after the test (x ± SD) Project 0 day 90 days P value Albumin (g/L) 36.94 ± 0.87 38.11 ± 1.19 0.3713 White ball ratio  1.12 ± 0.07  1.65 ± 0.30 0.1395 Triglyceride (mmol/L)  2.24 ± 0.77  2.10 ± 0.66 0.6378 High density lipoprotein  1.43 ± 0.47  1.46 ± 0.42 0.8386 (mmol/L) Low density lipoprotein  3.72 ± 0.71  3.86 ± 0.77 0.6312 (mmol/L)  6.29 ± 1.09  6.19 ± 1.13 0.8341 Total cholesrol (mmol/L) Fasting blood glucose (mmol/L)  8.00 ± 1.57  7.47 ± 1.43 0.3695

TABLE-US-00004 TABLE 4 Analysis of heart rate before and after the test (x ± SD) Project Number 0 day 90 days P value Heart rate 30 69.60 ± 11.59 73.23 ± 12.61 0.2579 (BPM)

[0156] There was no obvious abnormality in chest X-ray, electrocardiogram, abdominal ultrasound and urine routine examination before and 30 days after the test. During the test period the subjects had no adverse reactions or allergic reactions.

Test 1

[0157]

TABLE-US-00005 TABLE 5 Effective rate of the compound composition in improving aging symptoms, diseases of the elderly and sub-health symptoms Effective rate/% Improving diseases of the elderly and Improving aging sub-health Time symptoms symptoms Total 30 days 82.76% 73.08% 96.67% 60 days 86.67% 73.33%   100% 90 days 90.00% 86.67%   100%

[0158] It can be seen from the above Table 5 that the compound composition can improve the aging symptoms, diseases of the elderly and sub-health symptoms, and the effective rate increases gradually with the extension of taking time; after taking the compound composition for 60 days, the total effective rate of improving the aging symptoms can reach 100%, indicating that the compound composition of the disclosure has extremely significant effect on improving aging.

Test 2

[0159]

TABLE-US-00006 TABLE 6 Effective rate of the compound composition in improving skin condition of the elderly Effective rate/% improving skin improving improving elasticity collagen melanin Time abnormality abnormality abnormality 30 days 27.59% 20.69% 31.03% 60 days 27.83% 21.05% 30.95% 90 days 28.61% 22.17% 32.04%

[0160] The above table 6 shows that the compound composition can improve the skin elasticity, collagen and melanin abnormalities of the elderly to a certain extent, and the effective rate of improvement increases with the extension of taking time.

Test 3

[0161]

TABLE-US-00007 TABLE 7 Effect of the compound composition on improving anti-aging enzyme content in the elderly people content/% Superoxide Time dismutase Telomerase  0 day  16.90 ± 9.23    1.17 ± 1.05   30 days 22.11 ± 9.45*   1.59 ± 0.88*  60 days 26.57 ± 10.91** 1.64 ± 0.64*  90 days 29.08 ± 6.31**  1.72 ± 0.31** Note: compared with 0 day, *P <0.05; compared with 0 day, **P <0.01

[0162] In Table 7, the anti-aging enzyme content in the elderly people is detected, and it is found that the longer the compound composition was taken, the higher the contents of the superoxide dismutase and the telomerase am. The compound composition of the disclosure can effectively improve the anti-aging enzyme content in the elderly people.

TABLE-US-00008 TABLE 8 Effective rate of the compound composition in improving anti-aging enzyme in elderly people Effective rate/% Improving superoxide Improving Time dismutase Telomerase 30 days 86.67% 79.31% 60 days 88.30% 82.16% 90 days 90.08% 86.07%

[0163] It can be seen from the data in above Table 8 that the effective rate of the compound composition to improve the anti-aging enzyme in elderly people reached the highest on the 90th day of the test, and the effective rates of superoxide dismutase and telomerase were 90.08% and 86.07% respectively, indicating that the compound composition of the disclosure will gradually improve the effective rate of the anti-aging enzyme in elderly people with the extension of the taking time, which has a relatively good antioxidant effect.

Test 4

[0164]

TABLE-US-00009 TABLE 9 Effect of the compound composition on improving the content of vascular regulatory factors in elderly people Content/% Vascular endothelial Time Nitric oxide (NO) growth factor (VEGF)  0 day  21.80 ± 29.34   26.85 ± 27.27   30 days 47.17 ± 26.20** 47.87 ± 29.84** 60 days 50.27 ± 21.98** 54.08 ± 20.48** 90 days 53.08 ± 25.32** 57.29 ± 21.03** Note: compared with day 0 **P <0.01

[0165] It can be seen from the above table 9 that the contents of nitric oxide and vascular endothelial growth factor in the body of the subject, taking the compound composition by the disclosed method showed an upward trend, and increased with the extension of taking time, indicating that the compound composition has the effect of increasing the content of vascular regulatory factors in the body.

TABLE-US-00010 TABLE 10 Effective rate of the compound composition in improving vascular regulatory factors in elderly people Effective rate/% Improving Improving vascular Nitric endothelial gowth Time oxide (NO) factor (VEGF) Total 30 days 93.10% 93,10% 96.55% 60 days 95.24% 94.82% 98.99% 90 days 97.10% 96.04% 99.20%

[0166] It can be seen from the data in Table 10 above that the effective rate of compound composition in improving vascular regulatory factors in elderly people increased with the extension of taking time. On the 90th day, the total effective rate was 99.2% and the effective rates of NO and VEGF were 97.10% and 96.04% respectively. It is suggested that the compound composition has the functions of dilating blood vessels, promoting blood vessel regeneration and protecting cardio cerebral vessels.

[0167] Based on the analysis of the above results, it can be seen that the compound composition has a significant anti-aging effect and effectively improves the aging signs of the elderly, including aging, diseases and sub-health symptoms. The effective rate is high, with an overall effective rate of 100%. It can effectively improve the skin condition of the elderly, and enhance the contents of anti-aging enzyme and vascular regulatory factors in the elderly.

[0168] The above results show that the compound composition disclosed by the disclosure has anti-oxidation and anti-aging effects. The compound composition has no toxic side effects and is easy to be absorbed. It can be used to delay aging, repair DNA damage and anti-oxidation damage, and reduce the risk of geriatric diseases, which is suitable for market promotion and application.

[0169] Described above are merely illustrative of the disclosure to enable those skilled in the art to implement or use the disclosure, and ae not intended to limit the disclosure. It should be understood that any modification, replacement and change made by those skilled in the art without departing from the spirit of the disclosure should fall within the scope of the disclosure.