CN111513328A - Composition and health food containing cistanche deserticola extract and preparation method thereof - Google Patents
Composition and health food containing cistanche deserticola extract and preparation method thereof Download PDFInfo
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- CN111513328A CN111513328A CN202010307828.7A CN202010307828A CN111513328A CN 111513328 A CN111513328 A CN 111513328A CN 202010307828 A CN202010307828 A CN 202010307828A CN 111513328 A CN111513328 A CN 111513328A
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
- A23P10/00—Shaping or working of foodstuffs characterised by the products
- A23P10/30—Encapsulation of particles, e.g. foodstuff additives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Mycology (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Botany (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention provides a composition and a health food containing cistanche extracts and a preparation method thereof, wherein the composition containing the cistanche extracts is composed of the following components in parts by weight: 20-30 parts of cistanche extract, 18-25 parts of dogwood extract, 1-5 parts of semen cuscutae extract, 20-30 parts of rhizoma polygonati extract and 20-30 parts of cervus elaphus linnaeus. According to the invention, the cistanche extract, the dogwood extract, the semen cuscutae extract, the rhizoma polygonati extract and the cervus elaphus linnaeus are compatible according to a specific ratio, and the components play a synergistic effect, so that the obtained composition has the effects of relieving fatigue, accelerating the metabolic rate of lactic acid after exercise, increasing the tolerance of an organism and the like, has no toxic or side effect, and can be taken by consumers for a long time.
Description
Technical Field
The invention relates to the field of health-care food, in particular to a composition and health-care food containing cistanche extracts and a preparation method thereof.
Background
Modern life rhythm is accelerated, the pressure of work and life of people is increased, people in sub-health state are common, people in sub-health state for a long time can have various discomforts although no definite diseases exist, and the most common and obvious state is fatigue.
Fatigue is mainly divided into two major categories, physiological and pathological, wherein physiological fatigue includes 4 types of physical fatigue, mental fatigue, psychological fatigue and comprehensive fatigue. Physical fatigue is also called body fatigue because when a person moves for a long time and with high physical activity, a large amount of metabolites such as lactic acid, carbon dioxide, serum urea nitrogen and the like are generated in the body, and the substances are accumulated in the body to stimulate tissue cells and nervous systems of the body, so that the person can feel fatigue. Mental fatigue is caused by insufficient supply of blood and oxygen to the brain after long-time use of the brain, and particularly can cause symptoms such as inattention, dizziness, reaction retardation, limb weakness or somnolence and the like, and can cause a plurality of problems such as insomnia, dreaminess, nausea, vomiting, character change and the like. Mental fatigue is the most common and complex type of fatigue in modern life, and is caused by persistent stress and stress. The human is engaged in monotonous and mechanical work and learning activities for a long time, and the central local nerve cells are inhibited due to continuous tension, so that the enthusiasm and interest of the human in work on life are obviously reduced until tired emotion is generated. In the case of a person who suffers from psychological fatigue, mild persons may have symptoms of aversion, escape from work, learning, and life, and severe persons may also have symptoms of depression, neurasthenia, compulsive behavior, and phenomena such as smoking onset and alcoholism. The fatigue caused by modern life is not caused by a single reason, and is caused by physical and mental reasons, psychological and social reasons and possibly diseases, so that various 'symptoms' of single fatigue are not prominent and typical, and the fatigue caused by the non-single factor is called 'comprehensive fatigue'.
With the increasing development level of socio-economic and the increasing health awareness of people, more and more people realize the importance of relieving physical fatigue and improving sub-health status. Therefore, the invention aims to develop the health food suitable for sub-health people to take, and the health food can effectively relieve physical fatigue and improve the sub-health state.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a composition and a health food containing cistanche extracts and preparation methods thereof.
The invention aims to provide a composition containing cistanche extracts, which comprises the following components in parts by weight: 20-30 parts of cistanche extract, 18-25 parts of dogwood extract, 1-5 parts of semen cuscutae extract, 20-30 parts of rhizoma polygonati extract and 20-30 parts of cervus elaphus linnaeus.
According to the invention, the cistanche extract, the dogwood extract, the semen cuscutae extract, the rhizoma polygonati extract and the cervus elaphus linnaeus are compatible according to a specific ratio, and the components can play a synergistic effect, so that the obtained composition has the effects of relieving fatigue, accelerating the metabolic rate of lactic acid after exercise, increasing the tolerance of an organism and the like, has no toxic or side effect, and can be taken by consumers for a long time.
In a preferred embodiment of the present invention, the composition consists of the following components in parts by weight: 24 parts of cistanche extract, 20.1 parts of dogwood extract, 3 parts of semen cuscutae extract, 21.9 parts of rhizoma polygonati extract and 24.9 parts of cervus elaphus linnaeus.
Further, the cistanche extract is the sum of an alcohol extract and an aqueous extract of cistanche.
Preferably, the preparation method of the cistanche salsa extract comprises the following steps: extracting Cistanchis herba with 8 times of 70% ethanol for 2 times, each time for 2 hr, filtering, mixing extractive solutions, centrifuging at high speed, recovering ethanol from filtrate to relative density of 1.10-1.15 (50 deg.C), and concentrating to obtain concentrated solution; extracting the alcohol extraction dregs with 10 times of water for 2 times, each time for 1.5h, filtering, combining the extracting solutions, centrifuging at a high speed, combining the filtrate and the alcohol extraction concentrated solution, concentrating under reduced pressure to obtain an extract with the relative density of 1.15-1.20 (50 ℃), drying the extract under reduced pressure, crushing and sieving.
The cistanche deserticola is dry fleshy stem with scaly leaves of cistanche deserticola Cistanchestricola Y.C.Ma or cistanche tubulosa Cistanchebusosa (Schrenk) Wight of Orobanchaceae. The main active components of the cistanche deserticola extract comprise phenylethanoid glycosides, iridoid glycosides, lignin, amino acid, saccharides and the like.
Further, the dogwood extract is the sum of an alcohol extract and an aqueous extract of dogwood.
Preferably, the preparation method of the dogwood extract is the same as the preparation method of the cistanche extract.
The dogwood in the invention is dried mature pulp of dogwood (Cornusoficinalis Sieb. et Zucc.) belonging to Cornaceae. The main active ingredients of the dogwood extract comprise vitamins, minerals, amino acids, organic acids, dogwood saponins and the like.
Further, the dodder seed extract is an aqueous extract of dodder seeds.
Preferably, the preparation method of the semen cuscutae extract comprises the following steps: extracting semen cuscutae with 10 times of water for 2 times, each time for 1.5h, filtering, combining extracting solutions, centrifuging at high speed, concentrating the filtrate under reduced pressure to obtain an extract with the relative density of 1.15-1.20 (50 ℃), drying the extract under reduced pressure, crushing and sieving.
The dodder in the invention is dry mature seed of south dodder Cuscutaaustralisis R.Br. or dodder Cuscutachinensis Lam. of Convolvulaceae. The semen Cuscutae extract mainly contains semen Sojae Atricolor essence, flavone, sterols, polysaccharide, starch, protein, carotene, taraxanthin, organic acid, fatty acid, amino acids, amylase, etc.
Further, the rhizoma polygonati extract is an aqueous extract of rhizoma polygonati.
Preferably, the preparation method of the polygonatum sibiricum extract is the same as that of the dodder seed extract.
The rhizoma polygonati in the invention is dried rhizome of polygonatum kingianum (Polygonatum kingianum Coll. et Hemsl.) of Liliaceae, rhizoma polygonati (Polygonatum sibiricum Red.) or Polygonatum cyrtonema Hua. The rhizoma Polygonati extract mainly contains rhizoma Polygonati polysaccharide, steroid saponin, anthraquinone compounds, alkaloid, cardiac glycoside, lignan, vitamins and various amino acids useful for human body.
Further, the granularity of the cervus elaphus linnaeus is 200-1500 meshes. The cervus elaphus Linnaeus is a processed product of young horn of the ossified dense antler of male deer which is Cervus elaphus Linnaeus and belongs to the range of varieties allowed by China. The main active ingredients of cervus elaphus Linnaeus comprise estradiol, estrone, cholesterol, lecithin, cephalin, sphingomyelin, glycolipid and the like, wherein the total content of various amino acids reaches 50.13 percent, and the content of glycine is most abundant. The cervus elaphus linnaeus is a precious and fine medicinal material, is crushed into fine powder, and is used as raw powder, so that the full utilization of effective components is facilitated.
Furthermore, the content of total saponins (calculated by ginsenoside Re) in the composition is more than or equal to 0.55 percent, and the content of crude polysaccharide (calculated by glucose) is more than or equal to 0.30 percent.
The second purpose of the invention is to provide the application of the composition in preparing medicines or health-care foods with anti-fatigue efficacy.
The third purpose of the invention is to provide a health food containing cistanche extracts, which comprises the composition containing cistanche extracts and pharmaceutically acceptable auxiliary materials.
In a preferred embodiment of the present invention, the health food comprises, in weight percent: 70-80% of the composition containing cistanche deserticola extract, 7-9% of tricalcium phosphate, 0.5-1% of silicon dioxide, 0.8-1.2% of magnesium stearate and the balance of lactose.
Lactose is disaccharide composed of glucose and galactose, white crystal or crystalline powder, has sweetness about 70% of sucrose, is widely used in the fields of food and medicine production and manufacture due to its low hygroscopicity, and is used as diluent in the product. Lactose is decomposed into glucose and galactose by lactase in human intestinal tract, the glucose can directly participate in human metabolism to provide energy for human body, and 16.72 kilojoules of heat can be produced per gram of lactose.
Tricalcium phosphate is white, odorless and tasteless crystal or amorphous powder, food grade tricalcium phosphate is a food additive allowed by national laws, and tricalcium phosphate is used as an anticaking agent in the product to improve the quality stability of the finished product.
The food-grade silicon dioxide and the food-grade magnesium stearate are food additives which are allowed by China laws, and play a role in increasing material fluidity in food production. The two are compounded and then used as a lubricant in the product, so that the fluidity of the materials after total mixing is increased, and the subsequent production process is convenient to implement.
The invention also provides a preparation method of the health food, which comprises the following steps: sieving the components respectively, mixing the components uniformly according to the proportion, and preparing the obtained mixed powder into the required dosage form.
The health food can be prepared into common dosage forms such as capsules, tablets, pills and the like, preferably capsules, and not only reduces the hygroscopicity of each component and improves the stability of the product, but also is convenient to carry and take.
In a preferred embodiment of the present invention, the health food is prepared in the form of capsule, about 0.6g per capsule; the eating method and the eating amount are as follows: 2 times daily, 2 capsules each time, and is orally administered.
The invention has the beneficial effects that:
according to the invention, the cistanche extract, the dogwood extract, the semen cuscutae extract, the rhizoma polygonati extract and the cervus elaphus linnaeus are compatible according to a specific ratio, and the components play a synergistic effect, so that the obtained composition has the effects of relieving fatigue, accelerating the metabolic rate of lactic acid after exercise, increasing the tolerance of an organism and the like, has no toxic or side effect, and can be taken by consumers for a long time.
Detailed Description
The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention. The examples do not show the specific techniques or conditions, according to the technical or conditions described in the literature in the field, or according to the product specifications. The reagents or instruments used are conventional products available from regular distributors, not indicated by the manufacturer.
Example 1
The present invention provides a composition containing cistanche extracts, which comprises the following components: 120g of cistanche extract, 100g of dogwood extract, 15g of dodder extract, 110g of polygonatum extract and 125g of cervus elaphus linnaeus.
The embodiment also provides a preparation method of the composition, which specifically comprises the following steps:
(1) extracting Cistanchis herba with 8 times of 70% ethanol for 2 times, each time for 2 hr, filtering, mixing extractive solutions, centrifuging at 8000r/min, recovering ethanol from filtrate to relative density of 1.10-1.15 (50 deg.C), and concentrating; extracting the alcohol extraction residue with 10 times of water for 2 times, each time for 1.5h, filtering, combining the extracting solutions, centrifuging at a high speed of 8000r/min, combining the filtrate and the alcohol extraction concentrated solution, concentrating under reduced pressure (60-80 ℃ and-0.09-0.07 Mpa) to obtain an extract with the relative density of 1.15-1.20 (50 ℃), drying the extract under reduced pressure (60-80 ℃ and-0.09-0.07 Mpa), crushing, and sieving with a 80-mesh sieve to obtain a cistanche extract;
(2) taking dogwood as a raw material, and obtaining a dogwood extract by the same method as the step (1);
(3) extracting semen cuscutae with 10 times of water for 2 times, each time for 1.5h, filtering, combining extracting solutions, centrifuging at a high speed of 8000r/min, concentrating the filtrate under reduced pressure (60-80 ℃ and-0.09-0.07 Mpa) to obtain an extract with a relative density of 1.15-1.20 (50 ℃), drying the extract under reduced pressure (60-80 ℃ and-0.09-0.07 Mpa), crushing, and sieving with a 80-mesh sieve to obtain the semen cuscutae extract;
(4) taking rhizoma polygonati as a raw material, and obtaining a rhizoma polygonati extract by adopting the same method as the step (3);
(5) crushing the cervus elaphus linnaeus into particles of 200-1500 meshes by using an ultralow-temperature supersonic airflow crusher;
(6) the compositions are respectively weighed according to the mixture ratio and are evenly mixed to obtain the composition.
Example 2
The embodiment provides a health care capsule containing cistanche extract, which consists of the following components: 120g of cistanche extract, 100g of dogwood extract, 15g of dodder extract, 110g of polygonatum extract, 125g of cervus elaphus linnaeus, 48g of tricalcium phosphate, 6g of silicon dioxide, 6g of magnesium stearate and 70g of lactose.
The embodiment also provides a preparation method of the health-care capsule, which specifically comprises the following steps:
(1) extracting Cistanchis herba with 8 times of 70% ethanol for 2 times, each time for 2 hr, filtering, mixing extractive solutions, centrifuging at 8000r/min, recovering ethanol from filtrate to relative density of 1.10-1.15 (50 deg.C), and concentrating; extracting the alcohol extraction residue with 10 times of water for 2 times, each time for 1.5h, filtering, combining the extracting solutions, centrifuging at a high speed of 8000r/min, combining the filtrate and the alcohol extraction concentrated solution, concentrating under reduced pressure (60-80 ℃ and-0.09-0.07 Mpa) to obtain an extract with the relative density of 1.15-1.20 (50 ℃), drying the extract under reduced pressure (60-80 ℃ and-0.09-0.07 Mpa), crushing, and sieving with a 80-mesh sieve to obtain a cistanche extract;
(2) taking dogwood as a raw material, and obtaining a dogwood extract by the same method as the step (1);
(3) extracting semen cuscutae with 10 times of water for 2 times, each time for 1.5h, filtering, combining extracting solutions, centrifuging at a high speed of 8000r/min, concentrating the filtrate under reduced pressure (60-80 ℃ and-0.09-0.07 Mpa) to obtain an extract with a relative density of 1.15-1.20 (50 ℃), drying the extract under reduced pressure (60-80 ℃ and-0.09-0.07 Mpa), crushing, and sieving with a 80-mesh sieve to obtain the semen cuscutae extract;
(4) taking rhizoma polygonati as a raw material, and obtaining a rhizoma polygonati extract by adopting the same method as the step (3);
(5) crushing the cervus elaphus linnaeus into particles of 200-1500 meshes by using an ultralow-temperature supersonic airflow crusher;
(6) respectively sieving tricalcium phosphate, silicon dioxide and magnesium stearate with 80 mesh sieve; crushing lactose, and sieving with a 80-mesh sieve for later use;
(7) weighing the components according to the proportion, and mixing for 30min to obtain mixed powder;
(8) filling the mixed powder in a full-automatic capsule filling machine, wherein each capsule contains 0.6g of content;
(9) picking out unqualified capsules (malformed capsules and leaky capsules), polishing the qualified capsules in a polishing machine, and then conveying the polished qualified capsules to the next procedure for packaging.
Comparative example 1
The comparative example provides a health care capsule, which consists of the following components: 159g of cistanche deserticola extract, 146g of rhizoma polygonati extract, 165g of cervus elaphus linnaeus, 48g of tricalcium phosphate, 6g of silicon dioxide, 6g of magnesium stearate and 70g of lactose, and the preparation method is the same as the example 2.
Comparative example 2
The comparative example provides a health wine, which is prepared by the following steps:
weighing the Chinese medicinal raw materials: 500g of cordyceps sinensis, 500g of cistanche deserticola, 500g of cynomorium songaricum, 500g of epimedium herb, 800g of Chinese yam, 500g of morinda officinalis, 800g of astragalus membranaceus, 400g of prepared rehmannia root, 400g of semen cuscutae, 800g of sea buckthorn, 400g of prepared polygonum multiflorum, 400g of raspberry, 800g of cortex acanthopanacis, 400g of cervus elaphus linnaeus, 700g of glossy privet fruit, 700g of dogwood, 700g of cinnamon, 700g of black sesame, 700g of cassia seed, 700g of hawthorn, 700g of Chinese date, 700g of polygonatum odoratum, 700g of platycodon grandiflorum, 700g of wolfberry fruit, 700g of kudzu vine root, 700g of Chinese magnoliavine fruit, 500g of angelica, 400g of eucommia ulmoides, 600g of acanthopanax, 600g of safflower, 600g of cherokee rose fruit, 600g of Chinese chive seed, 600g of gynostemma pentaphylla, 400 g;
(2) loading the traditional Chinese medicine powder into a medicinal cloth bag per 1kg, and then soaking the medicinal cloth bag containing the traditional Chinese medicine powder of 1kg into 100kg of white spirit for at least 30 days to obtain a soaking solution;
(3) filtering the soaking solution, and removing the residue to obtain the health wine.
Test example 1 toxicological evaluation test for food safety
The reference standard is 'health food inspection and evaluation technical specification' (2003 edition)
First, acute toxicity test
Sample preparation: samples prepared according to the formulation and procedure described in example 2
Experimental animals: SPF-grade Kunming mouse
Mouse acute toxicity test (horns' method): each of the male and female mice has a weight of 18-21 g, and the mice are randomly divided into 5 groups according to the weight, wherein each group has 5 male and female mice, and the dosage is 0.46, 1.00, 2.15, 4.64 and 10.0g/kg.bw in sequence. After the animals in each group stop eating for 16 hours, the animals are continuously observed for 14 days after the animals are administrated by one-time oral gavage, and the toxic reaction and death condition of the animals in each group are recorded.
And (3) test results: as shown in Table 1, no obvious toxic reaction and no death case were found in each group of animals during the observation period, which indicates that the half lethal dose (acute toxicity) of the test substances to female and male mice is LD50Bw is more than 10.0g/kg, which is more than 100 times of the recommended dosage of human body. According to the acute toxicity grading standard, the tested substance belongs to an actual nontoxic substance.
TABLE 1 acute toxicity test results
Second, genotoxicity test
1. Sample preparation: samples prepared according to the formulation and procedure described in example 2
2. Test method
2.1 Ames test (plate incorporation method) with polychlorinated biphenyl (PCB) induced rat liver microsomal enzyme as in vitro metabolism activation system, identifying Salmonella typhimurium histidine-deficient mutant strain TA97, TA98, TA100, TA102 as qualified by his-, △ uvrB, rfa, R factor, PAQI plasmid, spontaneous regression, etc., culturing at 37 deg.C and 120rpm for 10 hours until the number of bacteria reaches 1 × 109And the cfu/mL is reserved. Adding 0.1ml test strain enrichment liquid, 0.1ml test substance and 0.5ml S9 mixed solution (a biochemical reagent, which plays the role of metabolism activation system in test and is induced by polychlorinated biphenyl inducer) into the top layer culture mediumThe treated liver homogenate of the rat is processed by 9000g of centrifugal force and assisted by auxiliary components, and is prepared by related subsequent processing processes), mixed evenly and poured onto a bottom layer culture medium, cultured for 48 hours at 37 ℃, and the colony number of each dish is counted. Spontaneous regression, blank control, solvent control and positive control were also set. Three parallel dishes were set for each dose group and control group and repeated once under the same conditions.
2.2 mouse bone marrow pleochromocyte micronucleus test: 50 Kunming mice are divided into 5 groups (5 females and 5 males in each group) according to the weight, wherein the weight of the Kunming mice is 25-30 g, and the mice are sequentially a negative control group, a positive control group (cyclophosphamide 40mg/kg.bw) and three dosage groups of 2.5, 5.0 and 10.0 g/kg.bw. Each group of mice was given test or control 2 times with 24-hour intervals by oral gavage. After 6 hours of the last gastric lavage, the test animals are killed by dislocation of cervical vertebrae, sternum marrow smears are taken, fixed by methanol, stained by Giemsa, and the ratio (PCE/NCE) of 200 pleochromophilic erythrocytes (PCE) to mature erythrocytes (NCE) is counted by oil-microscopy, and then the micronuclein number in 1000 pleochromophilic erythrocytes (PCE) is observed to calculate the micronuclein rate.
2.3 mouse teratospermia test: 25 Kunming male mice with weight of 25-30 g are randomly divided into 5 groups according to the weight, and the three groups are a negative control group, a positive control group (cyclophosphamide 40mg/kg.bw) and three dose groups of 2.5, 5.0 and 10.0 g/kg.bw. Each group of mice is orally administered with gastric lavage to a test object or a control object for 5 days continuously, then is continuously fed for 30 days, and then is subjected to cervical dislocation and sacrifice test animals, two-side epididymal smears are taken, methanol is fixed, eosin is stained, the number of various malformed sperms in each 1000 sperms is microscopically examined under a high power microscope, and the teratospermia rate is calculated.
2.4 result determination rule: in the three mutation-causing tests, the tested substance is abandoned for health care food when one or more tests are positive, and the next stage of toxicology test is carried out when only three tests are negative.
3. Test results
3.1 Ames test: in the two repeated detections of the Ames test plate doping method, the S9 activation system is added and not added at each concentration, the number of the recurrent colonies in each plate is within the normal value range, and does not exceed 2 times of the number of the spontaneous recurrent colonies, and no obvious measurement-effect relationship exists, which indicates that the Ames test result of the test material is negative and has no mutagenicity.
3.2 mouse bone marrow pleochromocyte micronucleus test: the PCE/NCE values of the three dose groups and the negative control group were within the normal range. The micronucleus rates of the three measurement groups are compared with those of the negative control group with the same property by the test of X2, and no significant difference (P is more than 0.05) exists, and no obvious measurement-effect relation is shown; but the difference is very significant compared with the positive control group of the same sex (P < 0.001). The results indicate that the mouse bone marrow micronucleus test for the test substance is negative.
3.3 mouse teratospermia test: the X2 test shows that the sperm aberration rate of the mice in the three-dose group is not significantly different (P is more than 0.05) compared with that in the negative control group, and the measurement-effect relationship or the multiple relationship relative to the negative control group is not shown; however, compared with the positive control group, the difference is very significant (P < 0.001). The results indicate that the test subject mouse has a negative teratospermia test.
And 3.4, the results of the three mutagenesis tests are negative, and finally, the genotoxicity test of the tested substance is judged to be negative, so that the next phase of toxicology test, namely a 30-day feeding test, can be carried out.
Feeding test for three and 30 days
1. Sample preparation: samples prepared according to the formulation and procedure described in example 2
2. The test method comprises the following steps: 80 SD rats with half male and female parts are adaptively fed for 5 days, and then are randomly divided into 4 groups (10 male and 10 female parts in each group) according to the body weight, and the groups are a normal control group and three dosage groups of 1.00, 2.00 and 4.00 g/kg.bw. The test substance is administered by feed incorporation. During the test period, each group of rats was kept in a single cage, fed freely and with water, continuously fed for 30 days, observed and recorded daily general performance, behavior, toxic performance, death condition, and feed addition and withdrawal, intake was settled weekly and body weight (including initial weight and final weight, wherein final weight is fasting), and weekly intake and food utilization were calculated. At the end of the test, the animals in each group stopped feeding at night, and after death after head breaking, the animals were roughly dissected, the organs were weighed and used as pathological sections of liver, kidney, stomach intestine, spleen, ovary and testis for examination in the control group and 100 times dosage group, and blood was taken for hemogram analysis and serum biochemical index measurement, and the total food intake and total food utilization rate were calculated.
3. Test results
During the test period, each group of animals do not find abnormal behavioral manifestations, and have no obvious toxic reaction, and none of the animals die; through analysis of variance, the change of the animal body weight of each dose group has no significant difference (P is more than 0.05) with that of a normal control group, which indicates that the tested organism has no obvious influence on the body weight and the growth and development of the long-term rat; through analysis of variance, the difference of the animals of each dosage group on the indexes such as weight increase, total food intake, food utilization rate and the like is not significant (P is more than 0.05) compared with the normal control group of the corresponding sex, which indicates that the food utilization rate of the rat is not influenced by adding the test substance; the biochemical indexes of the serum of the two control groups and the serum of each dosage group are within the normal value range, and the results of variance analysis and pairwise comparison show that the biochemical indexes of the serum of each dosage group have no significant difference compared with the two control groups of corresponding sex, which shows that the tested substance has no influence on the biochemical indexes of the serum of the rat; the hematology indexes of the two control groups and the dosage groups are within the normal value range, and the indexes among the groups have no significant difference, which indicates that the tested substance has no influence on the hematology indexes of the rat; the organ volume ratio indexes of the two control groups and each dosage group are within the normal range value range, and no significant difference exists between the groups, which indicates that the tested object has no influence on the organ coefficient of the rat; the appearance color and the size of the organs of the two control groups and the groups of each dosage are not abnormal, and obvious pathological changes such as exudation, hyperplasia, edema or atrophy and the like are not found, which indicates that the tested substance has no influence on the organs of the rat.
In conclusion, the test result of the 30-day feeding test of the test object is judged to be in accordance with the requirement.
Test example 2 evaluation test for health function
1. Sample preparation: samples prepared according to the formulation and procedure described in example 2;
comparative example 1 as control 1;
the comparative example 2 is used as a control group 2 (according to the relevant operation requirements of the pharmacological test of the liquid sample, the health care wine prepared in the comparative example 2 is concentrated);
the dose of the active ingredient actually administered to the test animals in comparative examples 1 and 2 was kept to be identical to the dose of the active ingredient administered to the test animals in the 20-fold dose group of example 2 during the test.
2. Test method
2.1 weight swimming test: adult male BALB/C mice 60 were randomly divided into 6 groups by body weight, which were a normal control group, three dose groups, control group 1 and control group 2, 10 mice per group. After 30 minutes from the last gavage, the subjects were loaded with 5% weight lead skin at the tail root, and the mice were placed in a swimming tank (water depth 40cm, water temperature 25 ℃. + -. 1 ℃) and the time from the start of swimming to death of the mice was recorded. During the test period, each group of mice had free access to food and water, and the body weights were weighed three times at the start, mid and final stages of the test, respectively.
2.2 serum Urea Nitrogen (BUN) test: the mice were continuously administered with the test substance or the control substance for 30 days (60 mice in total, 10 mice in each group, same-weight swimming test was measured), and after 30 minutes from the last administration of the test substance by gavage, orbital blood was taken after 20 minutes of swimming in a water tank at 30 ℃. + -. 1 ℃ and serum was separated and treated with a kit, and BUN was measured with a biochemical analyzer.
2.3 blood lactate accumulation and clearance assay: the test object or the control object is continuously given to the mouse for 30 days (total 60 mice, 10 mice in each group are measured and used for the same-weight swimming test), orbital blood is collected after the test object is given to the mouse through the last gastric lavage for 30 minutes, the blood-collected mouse is immediately taken out and wiped dry after swimming in a water tank at the temperature of 30 +/-1 ℃ for 10 minutes, the orbital blood is collected, and the swimming mouse is placed in a quiet state for resting for 20 minutes and then the orbital blood is again collected. After treatment with the kit, blood lactate data were measured before, after and after swimming for the mice.
2.4 liver glycogen assay: the test substance or the control substance is continuously administered to the mice for 30 days (total 60 mice, 10 mice in each group, the same-weight swimming test is measured), the test substance is administered to the mice after 30 minutes of the last gastric lavage, the liver is taken and rinsed by physiological saline, then the liver is sucked dry by filter paper, 75mg is accurately weighed, and the liver glycogen content is measured by an anthrone colorimetric method after the treatment by the kit.
2.5 test result judgment: (1) if the weight-bearing swimming time of the test sample group is obviously longer than that of the control group, and the difference is significant, the test result can be judged to be positive; (2) if the serum urea nitrogen of the test sample group is obviously higher than that of the control group and the difference is significant, the test result can be judged to be positive; (3) if the liver glycogen content of the test sample group is obviously higher than that of the control group, and the difference is significant, the test result can be judged to be positive; (4) if the area under the blood lactic acid curve of any test sample group is smaller than that of the control group and the difference is significant, the test result can be judged to be positive. The test result of the weight swimming is positive, and the test sample can be judged to have the function of relieving physical fatigue if any two indexes of three biochemical indexes of blood lactic acid, serum urea nitrogen and liver glycogen/muscle glycogen are positive.
3. Test results
3.1 weight swimming test: weighing the weight of the mouse in the beginning, the middle and the final stages of the test for three times, and after the last gastric lavage at the end of the test for 30min, putting the mouse in a constant-temperature swimming box for heavy swimming. The weight and weight swimming time are shown in table 2.
TABLE 2 weight of mice in each stage and influence of test object on swimming time
Note: indicates that the difference is significant compared with the normal control group (P < 0.05)
The body weight of each group of mice is increased during the test period, but no significant difference exists between the same group of mice, which indicates that the body weight of the mice is not obviously influenced by the tested substances. The result of the mouse weight swimming time shows that the weight swimming time of the mice of each dosage group is obviously increased compared with that of a control group, and the test object has the effect of prolonging the weight swimming time of the mice.
3.2 serum Urea Nitrogen (BUN) test: the results of the measurements of the body weight of the mice at each stage (initial, middle and final) and the serum urea nitrogen level after swimming during the test period are shown in Table 3.
TABLE 3 weight change of mice during serum Urea Nitrogen test and serum Urea Nitrogen level after 20min of constant temperature swimming
Note: indicates that the difference is significant compared with the normal control group (P < 0.05)
The weight of each group of mice is increased during the test period, but no significant difference exists between the same group of mice, which indicates that the weight of the mice is not obviously affected by the tested substances, and the result is consistent with the weight change condition of the load swimming test. The result of the serum urea nitrogen measurement after the swimming of the mouse shows that the urea nitrogen level of the mouse of each dosage group of the tested substance is lower than that of the normal control group, and the difference is significant. The results indicate that the test substance has the effect of reducing the serum BUN level of the mice after exercise.
Control 1 was lower than the normal control, but the difference was not significant; control 2 was lower than the normal control and the data difference was significant.
3.3 blood lactate accumulation and clearance assay: during the trial, the mice were weighed three times (initial, intermediate and terminal) as shown in table 4.
TABLE 4 change in body weight of mice during the blood lactate test
The body weight of each group of mice is increased during the test period, but no significant difference exists between the same group of mice, which indicates that the body weight of the mice is not obviously influenced by the tested substances, and the result is consistent with the results of the two previous tests.
Effect of test substances on blood lactic acid levels before and after swimming of mice: the blood lactic acid levels and their differences before, 10min after and 20min after swimming of the mice are shown in table 5.
TABLE 5 Effect of test substances on blood lactic acid levels before and after exercise in mice
Note: indicates that the difference is significant compared with the normal control group (P < 0.05)
As can be seen from the data in the table above, the blood lactic acid level of each group of mice increased significantly after exercise and decreased rapidly after 20min rest. Analysis of variance shows that the area under the blood lactic acid of the mice in the 20-time dose group after exercise is reduced compared with that of the normal control group, and the difference is significant, which indicates that the test object has the effect of reducing the blood lactic acid level of the mice after exercise.
3.4 liver glycogen assay: the results of the three (initial, intermediate and terminal) body weight and liver glycogen measurements of the mice during the test period are shown in table 6.
TABLE 6 Effect of test substances on mouse body weight and hepatic glycogen content
Note: indicates that the difference is significant compared with the normal control group (P < 0.05)
As can be seen from the data in the table above, the body weight of each group of mice increases during the test period, but no significant difference exists between the same group of mice, which indicates that the tested substances have no obvious influence on the body weight of the mice, and the result is consistent with the results of the first three tests. The result of liver glycogen determination shows that the difference of the 20 times dosage group is significant compared with the normal control group after the test object is given, and the result shows that the test object has the effect of increasing the liver glycogen content of mice; the control group 1 and the control group 2 have no significant difference compared with the normal control group.
In conclusion, the test results of the mouse weight swimming and the measurement of serum urea nitrogen after the mouse movement show that the weight swimming time of mice in the test object groups of 10 times, 20 times and 30 times of the test object groups is longer than that of the normal control group, the urea nitrogen level of the mice in each group is lower than that of the normal control group, and the difference is significant; the blood lactic acid test result shows that the area under the blood lactic acid curve of the mice in the 20-time dosage group is reduced compared with that of a normal control group after the mice move, and the difference is significant; liver glycogen measurement results show that the liver glycogen content of the 20-time dose group is higher than that of a normal control group, and the difference is significant. According to the regulation of 'health food inspection and evaluation technical specification', the tested substance is judged to have the effect of relieving physical fatigue.
The control group 1 has no significant difference compared with the normal control group in the serum urea nitrogen and the liver glycogen tests, and the control group 2 has no significant difference compared with the normal control group in the liver glycogen tests. It is shown that the products of comparative examples 1 and 2 are not as effective in fatigue resistance as the product of example 2 of the present invention.
Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (10)
1. The composition containing the cistanche deserticola extract is characterized by comprising the following components in parts by weight: 20-30 parts of cistanche extract, 18-25 parts of dogwood extract, 1-5 parts of semen cuscutae extract, 20-30 parts of rhizoma polygonati extract and 20-30 parts of cervus elaphus linnaeus.
2. The composition according to claim 1, which is characterized by consisting of the following components in parts by weight: 24 parts of cistanche extract, 20.1 parts of dogwood extract, 3 parts of semen cuscutae extract, 21.9 parts of rhizoma polygonati extract and 24.9 parts of cervus elaphus linnaeus.
3. The composition of claim 1 or 2, wherein the cistanche extract is the sum of an alcohol extract and an aqueous extract of cistanche;
and/or the dogwood extract is the sum of an alcohol extract and an aqueous extract of the dogwood;
and/or the dodder seed extract is an aqueous extract of the dodder seed;
and/or the rhizoma polygonati extract is an aqueous extract of rhizoma polygonati.
4. The composition of claim 3, wherein the cistanche extract or the cornus officinalis extract is prepared by a method comprising: extracting cistanche or dogwood with 8 times of 70% ethanol for 2 times, each time for 2 hours, filtering, combining extracting solutions, centrifuging, recovering ethanol from the filtrate until the relative density is 1.10-1.15, and keeping the concentrated solution for later use; extracting the alcohol extraction residues with 10 times of water for 2 times, each time for 1.5h, filtering, combining the extracting solutions, centrifuging, combining the filtrate and the alcohol extraction concentrated solution, concentrating under reduced pressure to obtain an extract with the relative density of 1.15-1.20, drying the extract under reduced pressure, crushing and sieving;
and/or the preparation method of the dodder seed extract or the polygonatum extract comprises the following steps: extracting semen cuscutae or rhizoma polygonati for 2 times by adding 10 times of water, each time for 1.5 hours, filtering, combining extracting solutions, centrifuging, concentrating the filtrate under reduced pressure to obtain an extract with the relative density of 1.15-1.20, drying the extract under reduced pressure, crushing and sieving.
5. The composition according to claim 1 or 2, wherein the composition has a total saponin content of 0.55% or more in terms of ginsenoside Re and a crude polysaccharide content of 0.30% or more in terms of glucose.
6. The composition as claimed in claim 1 or 2, wherein the cervus elaphus linnaeus is crude drug with a particle size of 200-1500 mesh.
7. Use of the composition of any one of claims 1 to 6 for the preparation of a medicament or health food having anti-fatigue effect.
8. A health food containing cistanche deserticola extract, which is characterized by comprising the composition containing cistanche deserticola extract as claimed in any one of claims 1-6 and pharmaceutically acceptable auxiliary materials.
9. The health food of claim 8, comprising, in weight percent: 70-80% of the composition containing the cistanche deserticola extract, 7-9% of tricalcium phosphate, 0.5-1% of silicon dioxide, 0.8-1.2% of magnesium stearate and the balance of lactose.
10. The method for preparing a health food according to claim 9, comprising: sieving the components respectively, mixing the components uniformly according to the proportion, and preparing the obtained mixed powder into the required dosage form.
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