CN100349530C - Health-care food with anti-fatigue and anti-oxidation function and its prepn. method - Google Patents

Abstract

The present invention relates to a health product with the function of resisting fatigue and oxidation and a preparation method thereof. The health product is an oral preparation mainly prepared from ginkgo leaf, desertliving cistanche, Chinese yam, active ingredient of ginseng and other auxiliary material. The present invention provides a health product by preparation technology of modern science of Chinese materia medica under the classical theory guide of the traditional Chinese medicine and pharmacy, and the health product has obvious effect of simultaneously relieving fatigue and resisting oxidation.

Description

A kind of health products and preparation method with antifatigue and anti-oxidation function

Technical field

The present invention relates to field of health care products.Be specifically related to a kind of health products and preparation method with antifatigue and anti-oxidation function.

Background technology

Development along with society, people's work pressure increases the weight of day by day, so wait in expectation can be among daily life careless for people always, can obtain not only alleviating physical fatigue simultaneously but also resist the health products with double health effect of autoxidation, to improve fitness, the quality of making the life better.

Summary of the invention

Technical problem to be solved by this invention is that under the classical theory of traditional chinese medicine instructs utilization modern Chinese herbal medicine length of schooling agent technology provides a kind of relieving fatigue and oxidation-resisting health-care product simultaneously.

Disclosed by the invention have the health products of antifatigue and anti-oxidation function by mainly containing the oral formulations that ginkgo leaf, saline cistanche, Chinese yam, genseng active component and other auxiliary materials are made.

Each active component of the present invention accounts for gross activity composition weight percent content: ginkgo leaf 16-32%, saline cistanche 32-56%, Chinese yam 16-32%, genseng 2-5%.

Other auxiliary materials of the present invention comprise various nutritional labelings, food additives and pharmaceutic adjuvant etc., as: vitamin E and purple perilla wet goods.

Another technical problem to be solved by this invention is to disclose above-mentioned preparation method with antifatigue and anti-oxidation function health products.

The preparation of the above-mentioned health products of the present invention comprises the following steps:

1. ginkgo leaf and Chinese yam, saline cistanche and genseng are divided into two groups, clean respectively, cut into slices or silk water and/or alcohol immersion, refluxing extraction;

2. above-mentioned two groups of extracts are merged centrifugation, supernatant concentration, spray-drying promptly gets mixed powder;

3. above-mentioned mixed powder is made various oral formulations according to a conventional method;

4. be not less than 2021mg/100g with general flavone content in the spectrophotometer detection preparation in rutin, thick polysaccharide is not less than 404.2mg/100g with glucose meter, and total saponin is not less than 853.5mg/100g in panaxoside glycosides Re.

Ginkgo leaf is one of primary raw material in health caring product prescription of the present invention, there is the experimental study report to set up animal experimental model by gavaging ginkgo biloba p.e, observes the vigor and the level of lipid of the antioxidase of its myocardial mitochondria with the mouse of an amount of swimming and tired swimming.After the mouse that gavages ginkgo biloba p.e passed through to swim in right amount and fatigue is swum, the vigor of antioxidase (SOD) all was significantly higher than control group separately, and the content of MDA (MDA) all significantly is lower than control group separately.Results suggest can alleviate the injury of the endogenous radical pair mouse that is produced by motion by taking ginkgo biloba p.e, and ginkgo biloba p.e has stronger Green Tea Extract damage and lipid-peroxidation effect, is natural free radical scavenger.

Many researchs show, and active oxygen in the body, free radical excessive meeting causing lipid peroxidation, protein denaturation, enzyme deactivation, polysaccharide degraded, the fracture of DNA chain, biological structure are impaired, cytoclasis and even body disease and death.Therefore, remove excessive active oxygen, free radical, its physiological significance is great.Ginko leaves flavone W and H have the stronger removing active oxygen and the effect of free radical, are the more antioxidants of a kind of function.

Saline cistanche is perennial parasitic herbaceous plant, and the sweet acid, warm in nature of distinguishing the flavor of has tonifying kidney and benefiting sperm, moisturizes functions such as laxation.Pharmacological evaluation shows that mouse swimming test is a kind of load physical exertion, sinks up to animal muscle power approach exhaustion.Experimental group raises the saline cistanche decocting liquid for the mouse mouth, can prolong the time-to-live of anoxic mouse, thereby saline cistanche has significant protective effect to heart and cerebral hypoxia.Illustrating that the saline cistanche decoction has builds up health, the effect of resisted motion fatigue, and the mouse swimming time is obviously prolonged.In the zoopery of the antioxidation of saline cistanche, adopt pyrogallol autoxidation, thiobarbituricacid (TBA) and fluorescence method, measure SOD activity in mouse core, liver, brain, the nephridial tissue, MDA and lipofuscin content respectively.The result: saline cistanche general glycoside GCS (125mg/kg, 250mg/kg) group can significantly improve SOD activity in brain, the nephridial tissue, and the 250mg/kg group improves the heart, SOD activities of liver, and significantly MDA and lipofuscin content are respectively organized in reduction.Results suggest GCS can improve the body tissue oxidation resistance effectively, prevents to organize lipid peroxidation injury.

Chinese yam is the Chinese medicine of using always, has the tonifying lung nourishing the stomach, a merit of the beneficial lung that promotes the production of body fluid, the puckery essence of kidney tonifying.Experimental results show that traditional Chinese medicine has greater advantage aspect removing interior free yl, the raising antioxidant ability of organism, existing many discovery, Chinese yam just has better antioxidation, Chinese yam polysaccharide can obviously improve SOD vigor and blood CAT vigor in the aging model mouse erythrocyte, improve the body antioxidation activity, suppress the formation of lipofuscin etc., make that the LPO level obviously reduces in aging model mouse blood, brain homogenate and the LH.The Chinese yam that this and tcm clinical practice are thought have making light of one's life by commiting suicide of tonifying middle-Jiao and Qi and ancient literature record prolong life wait act on consistent.Thereby the antioxidation of prompting Chinese yam polysaccharide may be the anti-ageing part mechanism of Chinese yam help.

Panaxoside is the important chemical composition of Chinese medicine genseng, and its hydrolysate comprises multiple sapogenins such as panoxadiol, panaxatriol, oleanolic acid.The antagonism of body is that the tired physiological mechanism that suppresses works during the positive motion load, and load is the principle of the physiological mechanism effect of fatigue recovery after stopping, and zoopery shows that genseng and compatibility thereof have the obvious anti-fatigue effect.There is experiment to show that genseng Rb saponins 50mg/Kg, 100mg/Kg, 200mg/Kg dosage group all can obviously dwindle its myocardial infarction area to acute myocardial infarction AMI 24h rat; reduce serum CK, LDH activity; and obviously reduce Serum LPO content; improve SOD, CAT and GSH-Px activity; can reach a conclusion: genseng Rb saponins has obvious protective effect to the rat acute myocardial infarction; may with its peroxidatic reaction of lipid that antioxidant radical is caused; strengthen activities of antioxidant enzymes in the body, the oxidative damage that reduces the radical pair cardiac muscle is relevant.Other experiment showed, that genseng can make the MDA in blood plasma and the tissue reduce, and erythrocytic SOD increased activity illustrates that genseng has antioxidation.

Antioxidant promotes the human immune system by the control free radical.Free radical is some unstable forms of oxygen, the more insulting important fat of meeting, thereby damage human body cell.Non-oxidizability materials such as vitamin E, C and A can be controlled these free radicals, the protection human immune system, thus can ease people psychology and physiological pressure.The MDA of small white mouse generated and raising SOD activity when vitamin E passed through to reduce the fatigability swimming exercise, and body is played certain protective role.In addition; the anti-oxidation function of vitamin E shows that vitamin E can be by self being oxidized to the fertility quinone; thereby ROO-is transformed into the inactive ROOH of chemical property; interrupt the chain reaction of lipid peroxidation; the peroxidation that effectively suppresses lipid, the protection cell is avoided the injury of the toxicant of unsaturated fat acid oxidase generation, plays an important role at aspects such as preventing aging, tumour; if in health products of the present invention, add vitamin E, then can further strengthen antifatigue and antioxidation.

In sum, more than fill a prescription, every flavor all has modern pharmacology basis and clinical research foundation, with the ginkgo leaf is monarch drug in a prescription, is that minister is assisted it with saline cistanche, Chinese yam, and assistant is with genseng, prescription meets traditional Chinese medical theory, and its preparation can reach alleviating physical fatigue and oxidation-resisting health-care function.

Can protect soft capsule to health products apricot spirit tablet power of the present invention and carry out the food safety evaluation on toxicology test, prove safety non-toxic:

1. acute oral toxicity test: the acute oral LD of SD rat and Kunming mouse ₅₀All, declare the genus non-toxic type greater than 15g/kg.bw.

2. genetic toxicity test: Salmonella reversion test, PCEMNR micronucleus test and three genetic toxicity test results of mouse sperm deformity test are all negative, show the no mutagenesis of this inspection product.

3. 30 days feeding trials of rat: in 30 days feeding trials of rat (maximum dose level is 100 times of human body recommended intake), tried the ANOMALOUS VARIATIONS that thing does not cause every important indicators such as rat holistic health, biochemical functions and organ-tissue morphology, maximum no-effect dose is greater than 3.3g/kg.bw (human body recommended intake 100 times) according to a preliminary estimate.

Can protect soft capsule to health products apricot spirit tablet power of the present invention and carry out the test of antifatigue anti-oxidation efficacy:

1. alleviating physical fatigue function animal experiment:

Apricot spirit tablet power can be protected soft capsule and be irritated stomaches by 10,20,30 times of per os of human body RD and gave animal subject 30 days.The result shows: apricot spirit tablet power can be protected soft capsule the mouse body weight is had no adverse effects: the 0.66g/kg.bw dosage group mouse swimming with a load attached to the body time significantly is longer than control group (P＜0.05); 1.00g/kg.bw dosage group Mouse Liver glycogen content is significantly higher than control group (P＜0.01), urea nitrogen content significantly is lower than control group (P＜0.05).The blood lactic acid content of three dosage group mouse swimming back 20min significantly is lower than control group (P＜0.05).By evaluation criterion, apricot spirit tablet power can be protected the effect that soft capsule has alleviating physical fatigue.

2. anti-oxidation function animal experiment

Adopt aged mouse to experimentize, establish three dosage groups of 0.17g/kg.bw, 0.34g/kg.bw and 0.68g/kg (be equivalent to respectively human body recommended intake 5,10 and 20 times), establish an aged contrast and young control group simultaneously.The result shows: the red blood cell MDA content that is tried object height dosage group significantly is lower than control group, and the vigor of GSH-Px is significantly higher than control group (P＞0.05) in the whole blood.By evaluation criterion, it is positive that apricot spirit tablet power can be protected soft capsule anti-oxidation function zoopery result.

3. anti-oxidation function human feeding trial:

Apricot spirit tablet power can be protected soft capsule antioxidation human feeding trial result and be shown that no abnormality seens such as duration of test experimenter's spirit, diet, sleep, stool and urine show that this inspection product have no adverse effects to human body.(P＜0.05) also was significantly higher than control group (P＜0.05) before the SOD activity of test group experimenter after test was significantly higher than test.Relatively do not have marked change before test-meal group test back MDA content and GSH-Px activity and the test, more also do not have significant difference with control group.By evaluation criterion, apricot spirit tablet power can be protected soft capsule anti-oxidation function.

The specific embodiment

Embodiment 1 alleviating physical fatigue function animal experiment

1. materials and methods

1.1. sample

1.1. apricot spirit tablet power can be protected soft capsule, the brown oil paste is provided by Xingling Sci. ﹠ Tech. Pharmaceutical Co., Ltd., Shanghai, and the human body RD is 2.0g/ people/day.

1.2. experimental animal and raising

ICR male white mouse, body weight are 18g-22g.

1.3. experimental technique

With body weight is that the ICR male white mouse of 18g-22g is divided into the blank group at random, three apricot spirit tablet power can be protected the soft capsule test group, the test group animal is tried thing by 10 times, 20 times, 30 times (0.33g/kg.bw, 0.66g/kg.bw, 1.0g/kg.bw) continuous irrigation stomaches of human body RD 2.0g/ day (0.033g/kg.bw) respectively, once a day, totally 30 days.The blank treated animal is irritated stomach with distilled water every day, and each treated animal is irritated stomach and carried out following experiment after 30 days respectively.

1.3.1. swimming with a load attached to the body experiment

After last gives given the test agent 30min, place the swimming case to swim the load mouse of 5% body weight sheet lead of root of the tail portion.The depth of water is no less than 30cm in the case, 25 ℃ ± 1 ℃ of water temperature, and the record mouse is from the extremely dead time of swimming beginning.

1.3.2. blood lactic acid is measured

After last was given sample 30min, it was not swimming with a load attached to the body 10min in 30 ℃ the water that animal is put temperature, and respectively adopted eyeball blood 20 μ L mensuration lactic acid content behind the rest 20min immediately before swimming, after the swimming.And calculate blood lactic acid TG-AUC as follows, to judge motion back blood lactic acid situation of change.

Blood lactic acid TG-AUC=5 * (blood lactic acid value before the swimming+3 * swimming back 0min blood lactic acid value+2 * swimming back rest 20min blood lactic acid value)

1.3.3. serum urea nitrogen determination

After last was given given the test agent 30min, it was not swimming with a load attached to the body 90min in 30 ℃ the water that mouse is put temperature, adopts eyeball blood 0.5mL behind the rest 60min, gets serum after the blood clotting and measures the serum urea nitrogen content with diacetyl-oxime method.

1.3.4. hepatic glycogen is measured

30min puts to death animal after last is given sample, gets liver and blots in the plate that is placed on the ice face with filter paper after the physiological saline rinsing, takes by weighing liver 100mg rapidly, measures hepatic glycogen content (anthrone method).

1.4. experimental data statistics

Adopt PEMS statistical software 3.0 editions that experimental data is carried out variance analysis, the heterogeneity of variance person uses rank test.

2. result

2.1. apricot spirit tablet power can be protected the influence of soft capsule to the mouse body weight: see Table 1-4.

Table 1-4 shows, apricot spirit tablet power can be protected the initial body weight of three dosage groups of soft capsule mouse, mid-term body weight and body weight and control group comparison during off-test, and there are no significant for difference (P＞0.05).

Table 1 apricot spirit tablet power can be protected the influence (swimming with a load attached to the body test group) of soft capsule to the mouse body weight

Group	Dosage (g/kg.bw)	Number of animals (only)	Initial body weight (g) (X ± S)	Body weight in mid-term (g) (X ± S)	End body weight (g) (X ± S)
						Control group	0	10	18.8±0.4	32.2±2.2	36.9±2.8
Low dose group	0.33	10	18.9±0.3	32.8±1.2	35.9±3.5
						Middle dosage group	0.66	10	18.8±0.4	32.4±1.8	35.8±1.9
High dose group	1.00	10	18.8±0.4	32.7±2.7	37.7±3.3

Table 2 apricot spirit tablet power can be protected the influence (blood lactic acid content mensuration group) of soft capsule to the mouse body weight

Group	Dosage (g/kg.bw)	Number of animals (only)	Initial body weight (g) (X ± S)	Body weight in mid-term (g) (X ± S)	End body weight (g) (X ± S)
						Control group	0	12	19.2±0.4	33.6±1.9	38.3±2.4
Low dose group	0.33	12	19.1±0.3	33.2±2.1	37.8±2.7
						Middle dosage group	0.66	12	19.2±0.4	32.4±2.7	35.7±2.9
High dose group	1.00	12	19.2±0.4	33.2±2.2	37.8±2.9

Table 3 apricot spirit tablet power can be protected the influence (hepatic glycogen content mensuration group) of soft capsule to the mouse body weight

Group	Dosage (g/kg.bw)	Number of animals (only)	Initial body weight (g) (X ± S)	Body weight in mid-term (g) (X ± S)	End body weight (g) (X ± S)
						Control group	0	12	20.3±0.5	34.7±1.6	39.8±1.5
Low dose group	0.33	12	20.2±0.4	34.1±2.1	38.6±2.3
						Middle dosage group	0.66	12	20.2±0.4	33.2±1.5	37.8±2.3
High dose group	1.00	12	20.2±0.4	34.2±1.7	39.4±2.2

Table 4 apricot spirit tablet power can be protected the influence (urea nitrogen content mensuration group) of soft capsule to the mouse body weight

Group	Dosage (g/kg.bw)	Number of animals (only)	Initial body weight (g) (X ± S)	Body weight in mid-term (g) (X ± S)	End body weight (g) (X ± S)
						Control group	0	11	21.2±0.4	34.8±2.1	38.8±2.2
Low dose group	0.33	11	21.3±0.5	33.5±3.2	37.7±2.6
						Middle dosage group	0.66	11	21.3±0.5	33.5±2.3	38.3±2.7
High dose group	1.00	11	21.2±0.4	35.3±1.8	39.8±2.5

2.2. apricot spirit tablet power can be protected soft capsule to the influence of mouse swimming with a load attached to the body time, the results are shown in Table 5.

Table 5 apricot spirit tablet power can be protected the influence of soft capsule to the mouse swimming with a load attached to the body time

Group	Dosage (g/kg.bw)	Number of animals (only)	The swimming with a load attached to the body time (s) (X ± S)
				Control group	0	10	191.5±51.1
Low dose group	0.33	10	255.6±78.1
				Middle dosage group	0.66	10	286.4±56.8 *

High dose group

1.00

10

242.1±70.5

^*Compare P＜0.05 with the photograph group

Show in the table 5 that the swimming with a load attached to the body time of each dosage group mouse of these inspection product all is longer than control group, there were significant differences (P＜0.05) for middle dosage group and control group comparison, and promptly apricot spirit tablet power can be protected soft capsule and can be prolonged the mouse swimming with a load attached to the body time.

2.3. apricot spirit tablet power can be protected the influence of soft capsule to mouse blood lactic acid content, sees Table 6.

Table 6 apricot spirit tablet power can be protected the influence of soft capsule to mouse blood lactic acid content

Group	Dosage (g/kg.bw)	Number of animals (only)	Blood lactic acid content (mmol/L) (X ± S)			Blood lactic acid TG-AUC
							Before the swimming	Swimming back 0min	Swimming back 20min
			Dosage group high dose group in the control group low dose group	0 0.33 0.66 1.00	12 12 12 12					3.35±1.19 3.73±1.76 2.83±1.69 2.92±1.55	5.05±1.44 4.93±1.35 4.26±0.87 4.75±1.50	2.54±1.09 1.66±0.58 * 1.58±0.38 * 1.52±0.48 *	117.83±32.48 109.23±27.22 93.77±18.24 101.06±32.41

^*Compare P＜0.05 with the photograph group

Before apricot spirit tablet power can be protected three dosage groups of soft capsule mouse swimming, swimming back 0min blood lactic acid content and three time point blood lactic acid TG-AUCs compare with control group, there are no significant for difference (P＞0.05), three dosage group mouse swimming back 20min blood lactic acid contents significantly are lower than control group (P＜0.01), be that apricot spirit tablet power can be protected soft capsule mouse swimming back 20min blood lactic acid content is had the reduction effect, but blood lactic acid TG-AUC is not had obvious effect.

2.4. apricot spirit tablet power can be protected the influence of soft capsule to the Mouse Liver glycogen content, sees Table 7.

Table 7 apricot spirit tablet power can be protected the influence of soft capsule to the Mouse Liver glycogen content

Group	Dosage (g/kg.bw)	Number of animals (only)	Hepatic glycogen content (mg/100g) (X ± S)
				Control group	0	12	1982.28±1114.77
Low dose group	0.33	12	1700.63±1251.83
				Middle dosage group	0.66	12	2483.41±646.84
High dose group	1.00	12	3567.00±1086.69 **

^*Compare P＜0.05 with control group

Table 7 shows that apricot spirit tablet power can be protected soft capsule 1.00g/kg.bw dosage group Mouse Liver glycogen content and is significantly higher than control group (P＜0.01).Be that apricot spirit tablet power can be protected soft capsule the Mouse Liver glycogen content is had the rising effect.

2.5. apricot spirit tablet power can be protected the influence of soft capsule to the mice serum urea nitrogen content, sees Table 8.

Table 8 apricot spirit tablet power can be protected the influence of soft capsule to the mice serum urea nitrogen content

Group	Dosage (g/kg.bw)	Number of animals (only)	Serum urea nitrogen content (mmol/L) (X ± S)
				Control group	0	11	8.28±1.04
Low dose group	0.33	11	9.25±1.11
				Middle dosage group	0.66	11	8.31±0.84
High dose group	1.00	11	7.34±0.85 *

^*Compare P＜0.05 with control group

Table 8 shows that apricot spirit tablet power can be protected soft capsule 1.00g/kg.bw dosage group mice serum urea nitrogen content and significantly is lower than control group (P＜0.05), and promptly this product (apricot spirit tablet power can be protected soft capsule) has the reduction effect to the mice serum urea nitrogen content.

3. brief summary

Apricot spirit tablet power can be protected soft capsule and be irritated stomaches by 10,20,30 times of per os of human body RD and gave animal subject 30 days.The result shows: apricot spirit tablet power can be protected soft capsule the mouse body weight is had no adverse effects; 0.66g/kg.bw the dosage group mouse swimming with a load attached to the body time significantly is longer than control group (P＜0.05); 1.00g/kg.bw dosage group Mouse Liver glycogen content is significantly higher than control group (P＜0.01), urea nitrogen content significantly is lower than control group (P＜0.05).Before the swimming of three dosage group mouse, swimming back 0min blood lactic acid content and three time point blood lactic acid TG-AUCs compare with control group, there are no significant for difference (P＞0.05), and three dosage group mouse swimming back 20min blood lactic acid contents significantly are lower than control group (P＜0.05).By evaluation criterion, apricot spirit tablet power can be protected the effect that soft capsule has alleviating physical fatigue.

Embodiment 2 anti-oxidation function animal experiments

1. material and method

1.1. sample

Apricot spirit tablet power can be protected soft capsule, lot number: 20030505, and the censorship product are the brown oil paste, are provided by Xingling Sci. ﹠ Tech. Pharmaceutical Co., Ltd., Shanghai.It is 0.5g * 4/ day/people that adult's taking dose is recommended by censorship side.Sample is given mouse stomach with edible vegetable oil preparation and dilution back during test.

1.2. animal used as test

Totally 50 of Kunming kind female mices, wherein 12 monthly ages 40 of above mouse, weight range 44-55 gram, few age in 8 weeks, mouse was 10, average weight 19.2 ± 1.0 grams.

1.3. dosage grouping

Can protect 5 times, 10 times and 20 times of soft capsule human body recommended amounts with apricot spirit tablet power and establish three dosage groups, i.e. 0.17g/kg.bw, 0.34g/kg.bw and 0.68g/kg.bw, other establish aged mouse control group and few age the mouse control group.Apricot spirit tablet power can be protected soft capsule and be prepared with vegetable oil, irritates stomach by 0.5% to animal.

1.4. experimental technique

To each dosage group mouse, irritate stomach by 0.5ml/100g.bw (promptly 100 gram body weight are irritated 0.5ml) volume to animal and tried thing every day, aged contrast and young group control animals all give edible vegetable oil by 0.5ml/100g.bw, continuous 40 days, build up MDA, SOD that bio-engineering research buys and GSH-Px kit specification by Nanjing during off-test and measure GSH-Px vigor in MDA content, SOD vigor and the whole blood in the animal erythrocyte.

1.5. experimental data statistics

Adopt the variance analysis of each experimental group and the comparison of control group mean that data are handled.Used software is PEMS3.0 " Chinese medicine encyclopedia Medical Statistics " statistical package (third edition).

2. result

2.1. apricot spirit tablet power can be protected the influence of soft capsule to the experimental mouse body weight

By table 9 as seen: at whole experimental session, aged mouse is respectively organized body weight does not all have significant difference (P＞0.05).

Table 9 apricot spirit tablet power can be protected the influence of soft capsule to each experimental stage body weight of mouse

Dosage (g/kg.bw)	The initial stage body weight		Mid-term body weight		Finish body weight
							Number of animals (only)	Body weight (g)	Number of animals (only)	Body weight (g)	Number of animals (only)	Body weight (g)
	Aged mouse contrasts 0.17 0.34 0.68	10 10 10 10	49.0±2.7 49.5±3.2 47.9±2.6 49.7±2.5	10 10 10 10	49.1±4.0 49.0±4.5 49.4±4.0 48.5±4.1	10 10 10 10							50.2±4.5 48.6±4.7 48.8±4.9 48.3±3.4

2.2. apricot spirit tablet power can be protected the influence of soft capsule to the red blood cell lipid peroxidation

Table 10 apricot spirit tablet power can be protected the influence of soft capsule to mouse red blood cell MDA content

Dosage (g/kg.bw)	Number of animals (only)	MDA content (nmol/ml)
			Lack mouse control group in age	10	5.4±0.66
Aged mouse control group	10	7.4±0.92 *
			0.17	10	6.94±0.93
0.34	10	6.80±0.92
			0.68	10	5.52±0.75 #

^*There were significant differences (P＜0.05) with few mouse control group comparison in age. ^#Relatively there were significant differences (P＜0.05) with aged mouse control group in expression.

Table 10 shows that aged mouse control group red blood cell MDA content is significantly higher than few mouse control group in age (P＜0.05), is tried object height dosage group (0.68g/kg.bw) and compares with aged mouse control group, and MDA content significantly reduces (P＜0.05).

2.3. apricot spirit tablet power can be protected the influence of soft capsule to the mouse red blood cell superoxide dismutase

Table 11 apricot spirit tablet power can be protected the influence of soft capsule to mouse red blood cell SOD

Dosage (g/kg.bw)	Number of animals (only)	SOD vigor (NU/ml)
			Lack mouse control group in age	10	3423.8±161.9
Aged mouse control group	10	3058.4±277.5 *
			0.17	10	3131.9±136.8
0.34	10	3102.1±167.7
			0.68	10	3154.3±214.3

^*There were significant differences (P＜0.05) with few mouse control group comparison in age.

By table 11 as seen, aged mouse control group erythrocyte sod vigor significantly is lower than few mouse control group in age (P＜0.05), is compared with aged mouse control group and try each dosage group of thing, and the SOD vigor does not have significantly and increases (P＞0.05).

2.4. apricot spirit tablet power can be protected the influence of soft capsule to mouse whole blood glutathione peroxidase

Table 12 apricot spirit tablet power can be protected the influence of soft capsule to the mouse blood GSH-Px

Dosage (g/kg.bw)	Number of animals (only)	The GSH-Px enzyme activity unit #
			Lack mouse control group in age	10	216.1±12.8
Aged mouse control group	10	196.9±12.7 *
			0.17	10	195.3±12.5
0.34	10	204.7±14.5
			0.68	10	213.2±15.7 #

^*There were significant differences (P＜0.05) with few mouse control group comparison in age. ^#Compare (P＜0.05=with aged mouse control group.

^#The enzyme activity unit definition: after the effect of regulation 1ml whole blood per minute deduction non-enzyme reaction 1g (GSH) reduced, the reduction value that makes 1g (GSH) was an enzyme activity unit.

Table 12 shows that aged mouse control group red blood cell GSH-Px vigor is significantly higher than few mouse control group in age (P＜0.05), is tried object height dosage group (0.68g/kg.bw) and compares with aged mouse control group, and the GSH-Px vigor significantly increases (P＜0.05).

3. conclusion

Apricot spirit tablet power can be protected the positive result of GSH-Px vigor in the MDA content and whole blood in the soft capsule anti-oxidation function animal experiment red blood cell, and by evaluation criterion, it is positive that apricot spirit tablet power can be protected the anti-oxidation function zoopery result of soft capsule.

Embodiment 3 anti-oxidation function human feeding trials

1. materials and methods

1.1. sample: apricot spirit tablet power can be protected soft capsule lot number is provided by Xingling Sci. ﹠ Tech. Pharmaceutical Co., Ltd., Shanghai: 20030505.Packing specification is 0.5g * 100/bottle, and the human body recommended intake is 2g/ people/day.

1.2. experimenter group: select to meet following standard person by the principle of voluntariness.

1.2.1. experimenter's choice criteria: select age in 45-65 year, physical condition is good, does not have obvious brain, the heart, liver, lung, kidney, blood illness, does not have the history of taking medicine for a long time, the elderly that aspiration is tried and guaranteed to cooperate.

1.2.2. experimenter's exclusion standard:

Pregnancy period or women breast-feeding their children; Be associated with serious disease patients such as cardiovascular, liver, kidney and hemopoietic system; Took the article person relevant in 30 days with being tried function; The person that do not take the given the test agent in accordance with regulations; Data is not judged effect or security person completely without method.

1.3. experimental design and grouping:

The experimenter is divided into test-meal group (50 people) and control group (52 people) at random, considers to influence result's principal element such as age, sex, living and diet custom etc. as far as possible, with the comparativity between the assurance group.Adopt two kinds of research methods of contrast between self cross-reference and group.

1.4. taking dose and time: oral 2 times of test-meal group everyone every day, each 2, took continuously 3 months.Control group is not done any processing.The duration of test experimenter does not change original life and eating habit, normal diet.

1.5. observation index

1.5.1. general situation: comprise experimenter's spirit, sleep, diet, stool and urine, blood pressure etc.

1.5.2. safety indexes: routine blood test, routine urinalysis, stool routine examination, the fluoroscopy of chest of X line, Abdominal B type ultrasonography, electrocardiogram, liver function, renal function etc.

1.5.3. effect index:

1.5.3.1. lipid peroxide content: the situation of change of MDA before and after the viewing test.Bio-engineering research institute's kit is built up in use Nanjing and recommend method is measured.

1.5.3.2. superoxide dismutase: the situation of change of SOD before and after the viewing test.Bio-engineering research institute's kit is built up in use Nanjing and recommend method is measured.

1.5.3.3. glutathione peroxidase: the situation of change of GSH-PX before and after the viewing test.Bio-engineering research institute's kit is built up in use Nanjing and recommend method is measured.

1.6. experimental data statistical analysis:

Use medical science encyclopedia statistical package (PEMS3.0) to carry out statistical analysis.The relatively employing paired t-test of difference before and after each index test.Test the relatively employing two sample mean t check of forward and backward test-meal group and control group group difference, adopt t ' check during heterogeneity of variance.

2. result

2.1. general situation: duration of test experimenter's spirit, diet, sleep, stool and urine etc. there is no unusually.

2.2. safety indexes observed result: heart rate, blood pressure, electrocardiogram before the test, the fluoroscopy of chest of X line, Abdominal B type ultrasonography, routine blood test, blood biochemistry index etc. are all in normal range (NR).Relatively there is no unusual before test back routine blood test and blood biochemistry index and the experiment.

2.3. effect index observing result:

2.3.1. lipid peroxide (MDA) content situation sees Table 13 before and after the test.

Test-meal group and control group comparison MDA content do not have significant difference before the test; Test-meal group test back does not have marked change with the preceding relatively MDA content of test, does not more also have significant difference with control group.

MDA content situation before and after table 13 test (X ± SD)

Project	Control group (N=52)		Test-meal group (N=50)
					Before the test	After the test	Before the test	After the test
	MDA(μ mol/ml)	1.75±0.86	1.83±0.58	1.84±0.68					1.77±0.31

2.3.2. superoxide dismutase (SOD) activity change situation sees Table 14 before and after the test.Test-meal group and control group compare the active there was no significant difference of SOD before the test; Significantly increase (P＜0.05) before test-meal group test back SOD activity and the test, more also there were significant differences (P＜0.05) with control group.

SOD activity change situation before and after table 14 test (X ± SD)

Project	Control group (N=52)		Test-meal group (N=50)
					Before the test	After the test	Before the test	After the test
	SOD(Nu/ml)	2490.6±389.6	2380.3±577.3	2431.7±476.8					2576.9±192.6 #*

Annotate: compare before and after self: ^#P＜0.01; Compare between group: ^*P＜0.01

2.3.2. glutathione peroxidase (GSH-Px) activity change situation sees Table 15 before and after the test.Test-meal group and control group compare the active no significant difference of GSH-Px before the test; Test-meal group test back and the active no marked change of the preceding relatively GSH-Px of test more also do not have significant difference with control group.

GSH-Px activity change situation before and after table 15 test (X ± SD)

Project	Control group (N=52)		Test-meal group (N=50)
					Before the test	After the test	Before the test	After the test
	GSH-Px (U/ml)	107.34±24.89	110.50±23.51	112.67±31.03					109.65±39.68

3. brief summary

Apricot spirit tablet power can be protected soft capsule antioxidation human feeding trial result and be shown, no abnormality seens such as duration of test experimenter's spirit, diet, sleep, stool and urine, and health check-up is no abnormality seen also, shows that preparation of the present invention has no adverse effects to human body; Before the SOD activity of test group experimenter after test is significantly higher than and tests (P＜0.05), also be significantly higher than control group (P＜0.05); Relatively do not have marked change before test-meal group test back MDA content and GSH-Px activity and the test, more also do not have significant difference with control group.By evaluation criterion, apricot spirit tablet power can be protected soft capsule anti-oxidation function.

Embodiment 4

Ginkgo leaf 555g

Saline cistanche 1110g

Chinese yam 800g

Genseng 64g

Vitamin E 16g

Perilla herb oil 540g

Beeswax 30g

Soybean lecithin 30g

1. traditional Chinese medicine extraction: ginkgo leaf and Chinese yam, saline cistanche and genseng divide two groups, through evaluation check (by the regulation of one one of Chinese Pharmacopoeia), respectively through cleaning, cut into slices or silk, weigh, drop in the multi-function extractor, add 70% ethanol of 12 times of amounts, soak after 2 hours, added hot reflux 1 hour, residue adds 10 times of water gagings and extracted 1 hour with method, merges extracted twice liquid;

2. tubular-bowl centrifuge: be used for high speed centrifugation (8000 rev/mins); It is 1.15 (60 ℃) that supernatant is evaporated to proportion;

3. spray-drying: soup carries out spray-drying, and its parameter is: moisture≤3.5% of 18000 rev/mins of rotating speeds, 185 ℃ of EATs, 85 ℃ of leaving air temps, powder delivery;

4. mix: mix after above-mentioned spray dried powder is sieved (200 order), get mixed powder 280g;

5. mixing: mix and powerful the stirring with liposoluble constituent (vitamin E, perilla herb oil, beeswax, soybean lecithin) in the matrix of soft capsule and the raw material, suspended matter (that is: content) 1000g;

6. sterilization treatment: above-mentioned suspended matter carries out microwave sterilization and handles microwave frequency 2450MHz, sterilization time 5min;

7. compacting: the suspendible powder that sterilization is good carries out the mensuration of functional component content, physics and chemistry, microorganism, records general flavone 2046mg/100g, total saponin 869.7mg/100g, and thick polysaccharide 415.7mg/100g makes soft capsule with above-mentioned content;

8. dry: as the soft capsule that is shaped to be carried out drying (≤60 ℃), get finished product, every 0.5g.

General flavone is measured in embodiment 5 preparations

1. rutin standard reference material solution: precision takes by weighing at the control substance of Rutin 15.0mg of 105 ℃ of drying under reduced pressure to constant weight, adds the methyl alcohol dissolving and is settled to 100mL, is made into 150 μ g/mL rutin standard liquids.

2. calibration curve

Accurate absorption standard rutin standard liquid 0.0,0.50,1.00,2.00,3.00,4.00mL are equivalent to rutin 0,75,150,300,450,600 μ g and move in the 10mL scale colorimetric cylinder, respectively add 30% ethanol to 5ml, respectively add 5%NaNO ₂Solution 0.3ml shakes up and places 5min, respectively adds 10%Al (NO ₃) ₃Solution 0.3ml shakes up, and places 6min again, adds 1.0mol/LNaOH solution 2ml, be settled to scale with 30% ethanol, shake up and place 10-20min, at 510nm wavelength place, make blank with zero pipe solution and measure absorbance respectively, concentration is returned the drawing standard curve with absorbance.

3. the preparation of need testing solution

Take by weighing the 1.0g sample, (embodiment 4 methods make content) packs tightly with filter paper, places boiling flask, adds 50-100mL70% ethanol, and after the infiltration, the 2h that refluxes under 80 ℃ of water-baths extracts substantially to flavone compound fully.After the crude extract cooling, decompress filter, and wash filter residue, merging filtrate with a small amount of 25% ethanolic solution.50 ℃ of following decompression distillation, remove ethanol wherein, solution is in flask does not have the alcohol flavor.Pour out solution in the flask, and divide the washing flask 3 times, behind the suction filtration, filtrate is poured in the separatory funnel, divide with the 75mL chloroform to extract degreasing 3 times with 30mL hot water, treat complete layering after, collect each time lower aqueous solution and be settled to 50mL.

Take by weighing 1-2g through pretreated polyamide powder, wet method dress post is used water saturation.Draw the aqueous solution 1-2mL after the above-mentioned degreasing, slowly splash in the post along chromatographic column, place certain hour, liquid to be measured is by after the abundant absorption, and with 70% ethanol or methanol-eluted fractions, flow velocity is 1.0mL/min, and liquid is colourless substantially to flowing out, and generally collects 10mL and gets final product.Above-mentioned eluate promptly can be used for after with the eluant, eluent constant volume measuring.

4. sample determination

The accurate sample extracting solution 1.00ml that draws carries out the mensuration of absorbance in the 510nm place by calibration curve preparation manipulation step in the 10ml colorimetric cylinder.

5. calculate

$X=\frac{m1\×V2}{m\×V2\×106}\×100=2046\mathrm{mg}/100g$

In the formula: X: content of total flavone in the sample (in rutin), mg/100g;

M1: the establishing criteria opisometer is calculated flavones content in the test solution, μ g;

M: the quality of sample, g;

V1: the volume that liquid branch to be measured is got, mL;

V2: the cumulative volume of liquid to be measured, mL.

Thick polysaccharide determination in embodiment 6 preparations

1. glucose titer: accurately take by weighing 1.0000g through 98-100 ℃ of glucose (AR) that is dried to constant weight, the back that is dissolved in water is diluted to 1000ml with water, and this solution 1ml contains 1mg glucose, with 10 times of preceding dilutions (0.1mg/ml), matching while using.

2. calibration curve

Precision pipettes glucose standard liquid 0,0.10,0.20,0.30,0.40,0.50,0.60mL, in the tool plug test tube that is placed in.Adding distil water is supplemented to volume 2.00mL, adds phenol solution 1.00mL again, shakes up, and adds concentrated sulfuric acid 5.00mL successively, shakes up, and puts boiling water bath 2min, takes out and puts room temperature, measures trap in wavelength 485nm place.With the sugar content is abscissa, and absorbance is an ordinate, the drawing standard curve.

3. the preparation of need testing solution

Sample extraction: take by weighing sample 2.0g, (embodiment 4 methods make content) places the 100mL volumetric flask, adds about water 80mL, on boiling water bath, heat 2h, be cooled to mend after the room temperature and add water to scale, behind the mixing, filter, discard filtrate just, collect remaining filtrate for the precipitation polysaccharide.

Accurately draw above whole filtrate 5.00mL, place the 50mL centrifuge tube, add absolute ethyl alcohol 20mL, behind the mixing 5min, with the centrifugal 5min of 3000r/min, abandoning supernatant.Residue is with milliliter washing of 80% (volume fraction) ethanolic solution number, centrifugal back abandoning supernatant, repeatable operation 3-4 time.The dissolving of residue water also is settled to 5.0mL, behind the mixing, for the precipitation glucan.

Accurately draw above whole solution 2.00mL, place the 20mL centrifuge tube, add 100g/L sodium hydroxide solution 2.0mL, copper test solution solution 2.0mL, boil heating 2min in the boiling water bath, cooling is with the centrifugal 5min of 3000r/min, abandoning supernatant.Residue is with milliliter washing of cleaning solution number, centrifugal back abandoning supernatant, and repeatable operation 3 times, residue dissolves with 10% (volume fraction) sulfuric acid solution 2.0mL and is transferred in the 50mL volumetric flask, and thin up is to scale, mixing.

4. sample determination

Draw need testing solution 1.00ml, add water and be supplemented to 2.00ml, add phenol solution 1.00ml again, shake up, add concentrated sulfuric acid 5.00ml successively, shake up, put boiling water bath 2min, take out and put room temperature, measure trap in wavelength 485nm place.

5. calculate

In the formula:

M: for measuring the amount of the suitable glucose of solution, μ g.

The mensuration of total saponin(e in embodiment 7 preparations

1. ginsenoside Re's standard liquid: accurately take by weighing ginsenoside Re's standard items 20.0mg, with the methyl alcohol dissolving and be settled to 10.0mL, promptly every 1mL contains ginsenoside Re 2.0mg.

2. sample treatment

Take by weighing sample 2.0g, (embodiment 4 methods make content) places the 100mL volumetric flask, adds about water 80mL, on boiling water bath, heat 2h, be cooled to mend after the room temperature and add water to scale, behind the mixing, carry out chromatography with PT-macroporous absorbent resin post, with the sample solution upper prop, wash post with 15mL, with water-solubility impurities such as sweetening offs, discard eluent, use the total saponin of 20mL85% ethanol elution again, collect eluent in evaporating dish, evaporate to dryness in water-bath.

Colour developing:

In the above-mentioned evaporating dish that has volatilized, accurately add 0.2mL5% vanillic aldehyde glacial acetic acid solution, rotate evaporating dish, make residual assorted dissolving, add 0.8mL perchloric acid again, move in the 10mL colorimetric cylinder behind the mixing, the jam-pack lid 15min that heats in the water-bath below 60 ℃ takes out, and the cooling back accurately adds glacial acetic acid 5.0mL, shake up the back with the 1.0cm cuvette in 560nm place and ginsenoside Re's standard pipe while colorimetric.

3. the drafting of calibration curve

Draw ginsenoside Re's standard liquid (2.0mg/mL) 0,20,40,60,80,100 μ L (being equivalent to ginsenoside Re 0,40,80,120,160,200 μ g), in the 10mL colorimetric cylinder, dry up, measure absorbance with the A1.3.2 development step with nitrogen.And drawing standard curve.

4. the result calculates

$X=\frac{m\×V_1\×100}{V_2\×1000}=869.7\mathrm{mg}/100g$

In the formula: X: total saponin(e (in the ginsenoside Re) in the sample, mg/100ml;

V ₁: sample extracting solution cumulative volume, ml;

V ₂: sample extracting solution is measured and is used volume, ml;

M: check in ginsenoside Re's amount the liquid to be measured, μ g from calibration curve.

Embodiment 8 preparation tablets

With dry ginkgo leaf 200g, Chinese yam 250g is one group, and saline cistanche 360g, genseng 40g are one group, adopts the preparation method with embodiment 4 to extract, and obtains spray dried powder.Getting spray dried powder 100g, add starch 100g low-substituted hydroxypropyl cellulose 40g, cross 80 mesh sieves respectively, mix, is that wetting agent is granulated with the Diluted Alcohol, dry whole grain, and it is a small amount of to add magnesium stearate lubricant, compressing tablet, every 0.3g.

The preparation of embodiment 9 oral liquids

Get the spray dried powder 100g that makes by embodiment 4 methods and add carboxymethyl cellulose 10g, asccharin 2g, Sodium Benzoate 2g is dissolved in the 8L distilled water, and it is an amount of to add essence, stirs, dissolves, filters packing, sterilization, packing is promptly.

Claims (3)

1, a kind of health products with antifatigue and anti-oxidation function, it is characterized in that described health products are to consist of with percentage by weight: ginkgo leaf 16-32%, saline cistanche 32-56%, Chinese yam 16-32% and genseng 2-5% are that pharmacodynamic raw materials is extracted the active component of acquisition and the oral formulations that other auxiliary materials are made, general flavone content is not less than 2021mg/100g in rutin in the preparation, thick polysaccharide is not less than 404.2mg/100g with glucose meter, and total saponin is not less than 853.5mg/100g in panaxoside glycosides Re;

Wherein pharmacodynamic raw materials ginkgo leaf and Chinese yam, saline cistanche and genseng are divided into two groups, difference water and/or alcohol immersion, and refluxing extraction obtains active component.

2, the health products with antifatigue and anti-oxidation function according to claim 1 is characterized in that wherein said other auxiliary materials are nutritional labeling, food additives and pharmaceutic adjuvant.

3, the preparation method with health products of antifatigue and anti-oxidation function according to claim 1 is characterized in that the preparation of these health products comprises the following steps:

1) ginkgo leaf and Chinese yam, saline cistanche and genseng are divided into two groups, clean respectively, cut into slices or silk water and/or alcohol immersion, refluxing extraction;

2) above-mentioned two groups of extracts are merged centrifugation, supernatant concentration, spray-drying promptly gets mixed powder;

3) above-mentioned mixed powder is made various oral formulations according to a conventional method;

4) be not less than 2021mg/100g with general flavone content in the spectrophotometer detection preparation in rutin, thick polysaccharide is not less than 404.2mg/100g with glucose meter, and total saponin is not less than 853.5mg/100g in panaxoside glycosides Re.