CN111387502A

CN111387502A - Propolis-bitter gourd composition with auxiliary blood sugar reducing effect and preparation method thereof

Info

Publication number: CN111387502A
Application number: CN202010414240.1A
Authority: CN
Inventors: 王鸿雁
Original assignee: Xi'an Royal Healthcare Products Co ltd
Current assignee: Xi'an Royal Healthcare Products Co ltd
Priority date: 2020-05-15
Filing date: 2020-05-15
Publication date: 2020-07-10

Abstract

The invention relates to a propolis balsam pear composition with auxiliary blood sugar reducing effect and a preparation method thereof, wherein the propolis balsam pear composition is prepared by taking propolis, balsam pear, fragrant solomonseal rhizome, a chromium source and a calcium source as raw materials and carrying out proper weight proportion, and the finally prepared propolis balsam pear composition with the auxiliary blood sugar reducing effect improves the degree of insulin secretion obstacle by conditioning pancreatic gland function, and finally achieves the effects of auxiliary blood sugar reducing and stable blood sugar reducing. The propolis balsam pear composition disclosed by the invention adopts medicinal and edible raw materials, reduces the dependence of a human body on medicaments, has small irritation, does not have toxic or side effect, and is safe and reliable. The propolis balsam pear composition disclosed by the invention can supplement a certain amount of calcium while assisting in reducing blood sugar, and is beneficial to improving the condition of calcium deficiency in vivo.

Description

Propolis-bitter gourd composition with auxiliary blood sugar reducing effect and preparation method thereof

Technical Field

The invention belongs to the technical field of health care products, and particularly relates to a propolis-bitter gourd composition with an auxiliary blood sugar reducing effect and a preparation method thereof.

Background

Diabetes is a group of metabolic diseases characterized by hyperglycemia. Hyperglycemia is caused by a defect in insulin secretion or an impaired biological action, or both. Hyperglycemia occurring in the long term of diabetes results in chronic damage to, and dysfunction of, various tissues, particularly the eyes, kidneys, heart, blood vessels, nerves.

Diabetes is one of three major diseases which affect human health most seriously at present, and clinical and disease research shows that diabetes is likely to become the disease which affects human health most seriously in the 21 st century, and poses serious threats to both the life quality and the life quality of patients. With the progress of urbanization and the change of consumption structure in the global range, in daily diet, the intake of more and more meat, protein and fat changes the diet structure of people mainly taking carbohydrate.

In the process of promoting urbanization process of each country, urban prevalence rate is continuously improved, and data show that about 4.63 million adults aged 20-79 years worldwide suffer from diabetes in 2019 (1 of 11 people is diabetic); it is predicted that by 2030, diabetics will reach 5.784 billion; it is expected that by 2045 years, diabetics will reach 7.002 billion.

1.1 hundred million diabetic patients and 4.9 hundred million diabetic patients in the early stage of diabetes in China mean that one diabetic person exists in every 10 diabetic patients, so that the development of the health-care food with the auxiliary blood sugar reducing function has great significance.

Disclosure of Invention

In order to solve the problems in the prior art, the invention provides a propolis balsam pear composition with an auxiliary blood sugar reducing effect and a preparation method thereof. The propolis balsam pear composition disclosed by the invention has the advantages that the degree of insulin secretion disorder is improved by conditioning the pancreatic function, and finally the effects of assisting in reducing blood sugar and stably reducing blood sugar are achieved. The propolis balsam pear composition disclosed by the invention adopts medicinal and edible raw materials, reduces the dependence of a human body on medicaments, has small irritation, does not have toxic or side effect, and is safe and reliable.

The technical scheme adopted by the invention is as follows:

a propolis balsam pear composition with auxiliary hypoglycemic effect comprises the following raw material components: 1000 portions of propolis, 4000 portions of balsam pear, 6000 portions of balsam pear, 4000 portions of polygonatum odoratum, 6000 portions of chromium source, 1-5 portions of chromium source and 1000 portions of calcium source, 1800 portions.

Further preferably, the propolis balsam pear composition with the auxiliary hypoglycemic effect comprises the following raw material components: 1400 parts of propolis, 5000 parts of balsam pear, 5000 parts of fragrant solomonseal rhizome, 3 parts of chromium source and 1400 parts of calcium source.

The chromium source is a trivalent chromium source.

The trivalent chromium source is one or a mixture of more of chromium picolinate, chromium yeast or chromium polynicotinate, and the calcium source is one or a mixture of more of pearl powder, calcium citrate, L-calcium threonate and L-calcium lactate.

The preparation method of the propolis balsam pear composition with the auxiliary hypoglycemic effect comprises the following steps:

(1) freeze drying propolis raw material, pulverizing, and sieving to obtain propolis fine powder;

(2) adding ethanol into the propolis fine powder for soaking and extracting to obtain a soaking solution;

(3) filtering the steeping liquor obtained in the step (2), and concentrating the filtrate to obtain propolis extract;

(4) mixing a calcium source with the propolis extract, drying in vacuum, and pulverizing at low temperature to obtain fine powder;

(5) crushing bitter gourd and polygonatum odoratum to obtain a mixed material; adding ethanol into the mixture for reflux extraction, and filtering to obtain an extract;

(6) concentrating the extracting solution obtained in the step (5) to obtain an extract;

(7) drying the extract obtained in the step (6) in vacuum, and crushing to obtain extract powder;

(8) taking a chromium source, and uniformly mixing the chromium source with the extract powder obtained in the step (7) to obtain a mixture;

(9) and (3) fully and uniformly mixing the fine powder in the step (4) and the mixture in the step (8), adding ethanol to prepare a soft material, and then sequentially granulating and drying to obtain the propolis balsam pear composition with the auxiliary blood sugar reducing effect.

In the step (1), the sieving is to pass through a sieve of 80-120 meshes;

in the step (2), the mass of the added ethanol is 3-5 times of the mass of the propolis fine powder.

In the step (2), the dipping extraction is repeated for three times, and the mass of the ethanol added each time is 3-5 times of the mass of the propolis fine powder;

the volume concentration of ethanol adopted in the first dipping extraction is 90%, and the time of the first dipping extraction is 6-8 h;

the volume concentration of ethanol adopted in the second dipping extraction is 85%, and the time of the second dipping extraction is 6-8 h;

the volume concentration of ethanol adopted in the third dipping extraction is 75%, and the time of the third dipping extraction is 6-8 h.

In the step (3), the relative density of the propolis extract is 1.35-1.40.

In the step (5), the reflux extraction is repeated for three times, and the ethanol extracted by each reflux extraction is 60-75% ethanol by volume concentration;

the amount of 60-75% ethanol added in the first reflux extraction is 8 times of the mass of the mixed material, and the time of the first reflux extraction is 2 hours; the amount of 60-75% ethanol added in the second reflux extraction is 6 times of the mass of the mixed material, and the time of the second reflux extraction is 1.5 h; the amount of 60-75% ethanol added in the third reflux extraction is 4 times of the mass of the mixed material, and the time of the third reflux extraction is 1 h;

in the step (6), the relative density of the extract is 1.15-1.20;

in the step (7), the granularity of the extract powder is 70-90 meshes.

In the step (9), the volume concentration of the ethanol is 88-92%, and the particle size after granulation is 18-22 meshes.

And (3) mixing the propolis balsam pear composition in the step (9) with a proper amount of conventional pharmaceutical excipients, and preparing pharmaceutically acceptable granules, pills, tablets or capsules with the auxiliary hypoglycemic effect according to a conventional forming process.

In order to facilitate understanding of the present invention, the raw materials and the drug effects of the present invention will be further described below.

Propolis, Propolis, a substance used by bees to repair and stick to the hive frame, is a solid gum with an aromatic odor. The substance is obtained from bee food, most of which is obtained by collecting gum from plant leaf bud, adding bee palate secretion and beeswax, and mixing. Propolis is a biochemically active substance. At present, the components identified by chemical household gas chromatography-mass spectrometry from propolis include flavone, terpenes, quinones, esters, alcohols, phenols, ethers, organic acids, and a large number of physiologically and pharmacologically active substances such as amino acids, enzymes, VB1, VB2, VB6, VE, VA, polysaccharides, and trace elements. Propolis has obvious blood-lowering effect. Researches show that flavonoid and terpene substances in the propolis have the effect of promoting synthesis of glycogen by utilizing exogenous glucose, and catalpol, pterostilbene and other substances in the substances have the obvious effect of reducing blood sugar. Propolis contains abundant trace elements, and research shows that zinc can maintain the structure and function of insulin; magnesium can be involved in pancreatic cell function regulation; potassium is an important component in cellular fluids and is involved in the metabolism of proteins, carbohydrates, etc. Propolis has no influence on normal mice fasting blood glucose, and has regulating effect on mice blood glucose increase and glucose tolerance decrease caused by alloxan.

The balsam pear, Momoradica charrantia L, is an annual herbaceous vine wood plant of the balsam pear genus of Cucurbitaceae family, and has bitter and cold nature, and has the efficacies of clearing away heat and toxic material, nourishing and strengthening body, reducing blood sugar and the like.

The drug is sweet in taste and mild in nature, and has the effects of improving the immunity of the organism, strengthening the heart, reducing blood fat and blood sugar, prolonging the life of animals and the like.pharmacological experiments prove that the dosage of 800mg/kg of the methanol extract of the polygonatum odoratum can reduce the blood sugar of STZ-induced hyperglycemia mice from 696 +/-60 mg/d L to 407 +/-35 mg/d L (P is less than 0.01). meanwhile, the drug can also inhibit the blood sugar of epinephrine-induced hyperglycemia mice.

Trivalent chromium is a micronutrient essential to human body and is an important component of Glucose Tolerance Factor (GTF). Chromium can activate insulin, improve glucose tolerance, and influence the secretion and release of insulin. In the absence of chromium, a human body can cause disorganization of glucose metabolism, increase of blood insulin, reduction of insulin affinity, weakening of B-cell sensitivity and reduction of the number of insulin receptors, and finally leads to abnormal glucose tolerance and diabetes. Chromium deficiency in human body is related to long-term refined food, high sugar diet and aging. In addition, chromium deficiency can occur as a result of strenuous physical exercise, physical trauma, pregnancy, and the like. The absorption of human body to organic chromium is better than that of inorganic chromium, and especially the absorption of chromium picolinate is the most ideal. The picolinic acid is an amino acid metabolite generated in the liver and kidney of human and mammals, is abundantly present in foods such as milk, and can be tightly combined with trivalent chromium to form chromium picolinate. The chromium picolinate has strong stability, can smoothly pass through cell membranes to directly act on cells, and has important auxiliary effect on the treatment of diabetes.

Ca, diabetic patients have low osteocalcin, i.e. low bone formation transformation rate and are easy to have osteoporosis, and the reasons are as follows: (1) the diabetic has the defects of metabolic disturbance of three nutrients caused by insufficient human insulin, and poor generation of bone matrix; (2) hyperglycemia increases urinary Ca excretion, indicating that calcium and phosphorus are lost from the kidneys, so diabetic patients are often accompanied by calcium deficiency, and severe patients develop osteoporosis. Thus, the propolis balsam pear compositions described herein incorporate a source of calcium.

The propolis balsam pear composition with the auxiliary blood sugar reducing function is prepared by adopting propolis, balsam pear, polygonatum odoratum, a chromium source and a calcium source as raw materials in a proper weight ratio, and the prepared propolis balsam pear composition with the auxiliary blood sugar reducing function improves the degree of insulin secretion obstacle by conditioning pancreas function based on a traditional Chinese medicine conditioning theory, and finally achieves the effects of auxiliary blood sugar reducing and stable blood sugar reducing. The propolis balsam pear composition disclosed by the invention adopts medicinal and edible raw materials, reduces the dependence of a human body on medicaments, has small irritation, does not have toxic or side effect, and is safe and reliable. The propolis balsam pear composition disclosed by the application can supplement a certain amount of calcium while assisting in reducing blood sugar, and is beneficial to improving the condition of insufficient calcium in a body.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the technical solutions of the present invention will be described in detail below. It is to be understood that the embodiments described are merely exemplary of the invention, and not of the invention in its entirety. All other embodiments, which can be derived by a person skilled in the art from the examples given herein without any inventive step, are within the scope of the present invention.

In the following examples 1g is represented by 1 part by weight.

Example 1

The embodiment provides a propolis-bitter gourd composition capsule with an auxiliary blood sugar reducing effect, which is prepared from the following raw materials:

1400 parts of propolis, 5000 parts of balsam pear, 5000 parts of fragrant solomonseal rhizome, 2.67 parts of chromium picolinate and 1400 parts of pearl powder.

The preparation method of the propolis-bitter gourd composition capsule with the auxiliary blood sugar reducing effect comprises the following steps:

(1) freeze drying propolis raw material, pulverizing, and sieving with 100 mesh sieve to obtain propolis fine powder;

(2) adding ethanol into the propolis fine powder, and sequentially performing three times of soaking extraction to obtain a soaking solution; the impregnation extraction specifically comprises the following steps: adding 90% ethanol with 4 times of mass of the propolis fine powder for primary soaking and extraction for 8h, and filtering; adding ethanol with volume concentration of 85% and volume of 4 times of the obtained filter residue, performing second impregnation extraction for 8h, and filtering; adding ethanol with volume concentration of 75% and volume of 4 times of the obtained filter residue, performing third immersion extraction for 8h, and filtering; combining the filtrates obtained by three times of soaking extraction to obtain the soaking solution;

(3) filtering the maceration extract obtained in step (2), and concentrating the filtrate under reduced pressure to obtain propolis extract with relative density of 1.35-1.40 (measured at 50 deg.C);

(4) mixing Margarita powder with the propolis extract, vacuum drying at 60 deg.C under 0.1Mpa, and pulverizing at below zero temperature to obtain fine powder;

(5) crushing bitter gourd and polygonatum odoratum to obtain a mixed material; adding ethanol into the mixture, and sequentially performing three times of reflux extraction to obtain an extracting solution; the reflux extraction specifically comprises the following steps: adding 8 times of 75% ethanol by mass into the mixed material, performing primary impregnation extraction for 2h, and filtering; adding ethanol with volume concentration of 75% 6 times the filter residue, performing second soaking extraction for 1.5 hr, and filtering; adding ethanol with volume concentration of 75% and volume of 4 times of the obtained filter residue, performing third immersion extraction for 1h, and filtering; combining the filtrates obtained by the three times of reflux extraction to obtain the extracting solution;

(6) concentrating the extracting solution obtained in the step (5) under reduced pressure to obtain an extract with the relative density of 1.15-1.20 (measured at 50 ℃);

(7) vacuum drying the extract obtained in the step (6), and crushing to obtain extract powder with the granularity of 80 meshes;

(8) taking chromium picolinate, and uniformly mixing the chromium source with the extract powder obtained in the step (7) to obtain a mixture;

(9) and (3) fully and uniformly mixing the fine powder in the step (4) and the mixture in the step (8), adding 90% ethanol in volume concentration to prepare a soft material, granulating by adopting a 20-mesh sieve, carrying out boiling drying, grading by using a 18-mesh sieve, and filling into a capsule shell to obtain the propolis balsam pear composition capsule with the auxiliary blood sugar reducing effect, wherein the mass of each capsule is 0.4 g. The capsule contains the following components in each 100g by detection: 1.01g of total soap, 1.79g of total flavone, 9.3g of calcium and 63mg of chromium picolinate. Taken twice a day, 3 granules at a time.

Example 2

1000 parts of propolis, 6000 parts of balsam pear, 4000 parts of polygonatum, 1 part of chromium picolinate and 1000 parts of pearl powder.

(1) freeze drying propolis raw material, pulverizing, and sieving with 80 mesh sieve to obtain propolis fine powder;

(2) adding ethanol into the propolis fine powder, and sequentially performing three times of soaking extraction to obtain a soaking solution; the impregnation extraction specifically comprises the following steps: adding 3 times of 90% ethanol by mass into the propolis fine powder, performing primary soaking extraction for 6h, and filtering; adding ethanol with volume concentration of 85% and volume of 3 times of the obtained filter residue, performing second impregnation extraction for 6h, and filtering; adding ethanol with volume concentration of 75% and volume 3 times of the obtained filter residue, performing third immersion extraction for 6h, and filtering; combining the filtrates obtained by three times of soaking extraction to obtain the soaking solution;

(4) mixing Margarita powder with the propolis extract, vacuum drying at 50 deg.C under 0.05Mpa, and pulverizing at 10 deg.C or below to obtain fine powder;

(5) crushing bitter gourd and polygonatum odoratum to obtain a mixed material; adding ethanol into the mixture, and sequentially performing three times of reflux extraction to obtain an extracting solution; the reflux extraction specifically comprises the following steps: adding 8 times of ethanol with volume concentration of 60% by mass into the mixed material, performing primary impregnation extraction for 2h, and filtering; adding ethanol with volume concentration of 60% 6 times the filter residue, performing second soaking extraction for 1.5 hr, and filtering; adding ethanol with volume concentration of 60% and volume of 4 times of the obtained filter residue, performing third immersion extraction for 1h, and filtering; combining the filtrates obtained by the three times of reflux extraction to obtain the extracting solution;

(7) vacuum drying the extract obtained in the step (6), and crushing to obtain extract powder with the granularity of 70 meshes;

(9) and (3) fully and uniformly mixing the fine powder in the step (4) and the mixture in the step (8), adding 88% ethanol in volume concentration to prepare a soft material, granulating by using a 18-mesh sieve, carrying out boiling drying, grading by using the 18-mesh sieve, and filling into a capsule shell to obtain the propolis balsam pear composition capsule with the auxiliary blood sugar reducing effect, wherein the mass of each capsule is 0.4 g.

Example 3

1800 parts of propolis, 4000 parts of balsam pear, 6000 parts of polygonatum, 5 parts of chromium picolinate and 1800 parts of pearl powder.

(1) freeze drying propolis raw material, pulverizing, and sieving with 120 mesh sieve to obtain propolis fine powder;

(2) adding ethanol into the propolis fine powder, and sequentially performing three times of soaking extraction to obtain a soaking solution; the impregnation extraction specifically comprises the following steps: adding 90% ethanol with 5 times of mass of the propolis fine powder for primary soaking and extraction for 8h, and filtering; adding ethanol with volume concentration of 85% and volume of 5 times of the obtained filter residue, performing second impregnation extraction for 8h, and filtering; adding ethanol with volume concentration of 75% and volume 5 times of the obtained filter residue, performing third immersion extraction for 8h, and filtering; combining the filtrates obtained by three times of soaking extraction to obtain the soaking solution;

(4) mixing Margarita powder with the propolis extract, vacuum drying at 80 deg.C under 0.2Mpa, and pulverizing at below zero temperature to obtain fine powder;

(7) vacuum drying the extract obtained in the step (6), and crushing to obtain extract powder with the granularity of 90 meshes;

(9) and (3) fully and uniformly mixing the fine powder in the step (4) and the mixture in the step (8), adding 92% ethanol in volume concentration to prepare a soft material, granulating by adopting a 22-mesh sieve, carrying out boiling drying, grading by using a 18-mesh sieve, and filling into a capsule shell to obtain the propolis balsam pear composition capsule with the auxiliary blood sugar reducing effect, wherein the mass of each capsule is 0.4 g.

Examples of the experiments

Firstly, safety toxicology test:

1. description of the samples:

the sample was the capsule described in example 1, the content of the capsule was brown powder, and the daily intake was 2.40g/60kg · bw. The sample solvent was distilled water.

2. Experimental animals and mode of exposure:

ordinary Kunming mouse and Wistar rat bred in the experimental animal center of Chinese medical university are selected, and the experimental animal produces license number SCXK (Liao) 2002-. The block mouse feed is provided by Shenyang city in flood zone animal experiment feed factories (Liao Shi moving Chinese character 'huan' 1999-024), except for 30 days, the feeding experiment adopts a feed gradual amplification mixing method, other animal experiments adopt a tube feeding method, the stomach filling amount of rats is 10 ml/kg-bw, and the stomach filling amount of mice is 20 ml/kg-bw.

3. The test conditions are as follows:

the barrier system animal laboratory is constant in temperature and humidity, the temperature is 22 +/-1.5 ℃, the temperature is 50% +/-10%, the working illumination is 160-.

4. Equipment reagent:

AE L-160 type electronic balance (sensory 0.0001g, Mechan M112289), ES-2000A type electronic balance (sensory 0.01g, Liaojie 02041503103), biological microscope, full-automatic blood cell counter (COBASET Shen Yi No. P120208), full-automatic biochemical analyzer (COBASS MIRA, Shen Yi No. P120207), centrifuge, head cutter, microtome, full-automatic dehydrator, dissecting scissors, tweezers, and various biochemical kits.

5. Acute toxicity test in mice (one-time maximal method):

20 healthy Kunming mice, 10 male and female respectively, were selected for testing. The weight of the mouse is 18-22 g. After the gavage was performed at a dose of 10.00g/kg · bw, the gavage was continuously observed for 14 days, and test records were made.

6. Acute toxicity test in rats (one-time maximal method):

healthy Wistar rats, 20 in number and 10 in number, respectively, were selected for testing. The weight of the rat is 180-220 g. After the gavage is carried out at the dose of 10.00g/kg · bw (gavage is carried out for 2 times at intervals of 4h, and the dose of each time is 5.00g/kg · bw), the continuous observation is carried out for 14 days, and the test record is made.

7. Contaminant mutagenicity assay (Ames assay):

the test is carried out by four test strains of the salmonella typhimurium histidine-deficient strain TA97, TA98, TA100 and TA102 which are identified to meet the requirements. Rat liver homogenate induced by polychlorinated biphenyls (PCB) was used as an in vitro metabolic activation system. Based on the toxicity assay results, the test samples were assigned to 5 dose groups of 0.05, 0.10, 0.50, 2.50, 5.00 mg/dish, along with spontaneous regression and positive controls. 0.1ml of test seedling enrichment liquid, 0.1ml of test sample solution and 0.5ml of S-9 mixed solution (when metabolic activation is needed) are added into top agar, mixed uniformly and poured onto a bottom culture medium plate, and 3 plates are used for each dose. The cells were incubated at 37. + -. 1 ℃ for 48h and the number of colonies transformed per dish was counted. A test sample is judged to be positive if it has a dose-response relationship in which the number of recurrent colonies is 2 times or more as large as the number of spontaneous recurrent colonies. The whole set of experiments was repeated twice under the same conditions.

8. Mouse marrow pleochromocyte micronucleus test:

the test was performed by oral gavage with two 24h intervals. 50 mice with a weight of 25-30g were randomly divided into 5 groups of 10 mice each with half of the males and females. The cyclophosphamide dosage of 40mg/kg · bw is used as a positive control, the sample solvent is used as a negative control, and the samples for the test are set to 1.25, 2.50 and 5.00g/kg · bw3 dosage groups and are prepared to the required concentration by the sample solvent. After 6h of the last administration of the test sample, the animals were sacrificed by cervical dislocation, and the sternal bone marrow was diluted with calf serum and smeared, fixed with methanol, and stained with Giemsa. Under light microscopy, 1000 pleochromophilic erythrocytes (PRC) were counted per animal, the micronucleus incidence was measured in micronucleus-containing PRC, and statistical treatment was performed.

9. Mouse teratospermia test:

25 sexually mature male mice weighing 25-30g were used and randomly divided into 5 groups. The cyclophosphamide dosage of 40mg/kg · bw is used as a positive control, the sample solvent is used as a negative control, and the samples for the test are set to 1.25, 2.50 and 5.00g/kg · bw3 dosage groups and are prepared to the required concentration by the sample solvent. Gavage is performed once a day for 5 consecutive days, animals are sacrificed 30 days after the last gavage, epididymis is prepared and stained by eosin, 5 animals are counted in each group, 1000 sperm with complete structure are counted in each animal, the incidence rate (in percentage) of abnormal sperm is calculated, and statistical processing is performed.

10. Rat 30-day feeding trial:

weaned rats (75-90 g body weight at the start of the test, not more than. + -. 20% of the mean body weight) were used in 80 animals, male and female. The test results were randomly divided into 125, 2.50 and 5.00g/kg · bw3 dose groups and basal feed control groups. The dosages of each group were converted to the amount of feed (mg/kg) per 100g of body weight, and 10% of body weight. Feeding in a single cage, weighing the weight and the food input amount once a week, and calculating the food utilization rate; at the end of the test, the measurement of hemoglobin, red blood cell count, white blood cell count and classification, platelet count, serum glutamic-pyruvic transaminase, glutamic-oxaloacetic transaminase, urea nitrogen, creatinine, glucose, serum albumin/globulin, total cholesterol, triglyceride, high density lipoprotein cholesterol, etc.; the absolute weights of the organs of liver, kidney, spleen, testis, ovary were weighed and the organ volume ratio was calculated. When the gross examination of animals in each dose group shows no obvious lesion, the histopathological examination of liver, kidney, stomach, intestine, spleen, testis and ovary of the high dose group and the control group is carried out, and the statistical treatment is carried out.

11. Data processing:

in the measurement data, the average comparison of multiple samples adopts one-factor variance analysis, the minimum significant difference method, new repolarization difference method and two-sample average t test are simultaneously adopted for the comparison of each experimental group and a solvent control group, the homogeneity test of multiple sample variance (Bartlett method) and the homogeneity test of two-sample variance of each experimental group and the solvent control group are simultaneously carried out, if the variance is not uniform, the variable conversion is carried out, the sum test (Wilcoxon two-sample comparison method) is adopted, the counting data adopts a row × list x²Testing, comparing each experimental group with solvent control group by adopting four-grid table x²Checking for non-compliance with the four-grid table x²The data on the test conditions were obtained by the rank-sum test (Wilcoxon two-sample comparison). The original data of each item of inspection is automatically subjected to statistical analysis in an Excel 2002 software program, and the related data obtained by analysis is automatically transmitted to the designated position of an inspection report form.

12. Experimental unit: liaoning disease prevention and control center

13. The experiment summary:

(1) acute toxicity test:

① mouse oral acute toxicity test, the test sample is gavaged at a dose of 10.00 g/kg-bw for mice of two sexes, no obvious toxic manifestation is observed and no death is caused after 14 days of observation, the oral acute toxicity L D50 of the test sample to the mice of two sexes is respectively more than 10.00 g/kg-bw and is classified according to the acute toxicity, and the test sample belongs to a practical nontoxic substance.

② oral acute toxicity test in rats:

the test sample is gavaged at a dose of 10.00g/kg · bw for rats of two sexes, no obvious toxic manifestation and no death are observed after 14 days of observation, the oral acute toxicity L D50 of the test sample to the rats of the two sexes is larger than 10.00g/kg · bw. according to the acute toxicity classification, and the test sample belongs to a practical nontoxic substance.

The oral acute toxicity (L D50) of the test sample to mice of two sexes is more than 10g/kg bw, the oral acute toxicity (L D50) of the rats of two sexes is more than 10g/kg bw, and the test sample is classified according to the acute toxicity and is practically nontoxic.

(2) And (3) genetic toxicity test:

① contaminant mutagenicity detection (Ames test):

the number of the mutator colonies of each dose group of the sample for the test does not exceed the number of the mutator colonies of the solvent control group by more than 2 times, and the test has no dose-reaction relation, and the results are negative when a liver microsome enzyme activation system is added or not added to four test strains of salmonella typhimurium TA97, TA98, TA100 and TA102, and the test results can be repeated.

② mouse bone marrow pleochromocyte micronucleus test:

compared with the sample solvent control group, the incidence rate of bone marrow polystaining erythrocyte micronucleus of mice with male and female sex of each dose group of the sample for the test is not poor (P is more than 0.05); compared with the sample solvent control group, the cyclophosphamide group has significant difference (P is less than 0.05). No obvious damage effect of the test sample on bone marrow cell chromosomes of mice with male and female sexes is found.

③ mouse teratospermia test:

the mouse teratospermia incidence rate of each dosage group of the test sample is compared with that of a sample solvent control group, and no significant difference exists (P is more than 0.05); compared with a sample solvent group, the teratospermia incidence rate of the mice of the cyclophosphamide group has significant difference (P is less than 0.05), and no obvious damage effect of a test sample on germ cells of male mice is seen.

The result shows that the sample for the test has no genotoxic effect.

(3) Rat 30-day feeding trial:

① clinical examination, organ weighing, blood biochemistry examination, and hematology examination results:

compared with the control group, the experimental group and the control group in each test item have no significant difference (P is more than 0.05). During the test period, the rats move freely, hair is bright, and water, food and urine are normal.

② histopathological examination results:

a. general inspection:

the co-examination experiment showed 80 rats, each half male and female. After the autopsy, no obvious abnormality is observed in the color, shape, size and the like of the main organs such as heart, lung, liver, spleen, kidney, stomach, intestine, testis (ovary), brain and the like.

b. And (3) microscopic examination:

the high dose experimental group and the control group were examined for 40 rats, male and female halves.

Liver: the capsule is complete, the structure of hepatic lobule is clear, the hepatic cell cord is arranged regularly, the hepatic cells have no obvious degeneration, necrosis and other changes, and inflammatory cell infiltration is occasionally seen in a local manifold area. Wherein, the female rats in the control group have 2 local areas with inflammatory cell infiltration; in the high dose experimental group, inflammatory cell infiltration was observed in 1 male rat and 2 local zone of the assembled female rat.

Kidney: glomeruli, renal capsule and renal tubule have normal structures, and inflammatory cell infiltration is occasionally seen in local renal interstitium. Wherein, the male rats in the control group have 1 local renal interstitial inflammatory cell infiltration; in the high-dose experimental group, 1 male rat has mild extravasated blood in local renal interstitial vessels.

Gastrointestinal tract: the rats in the control group and the high-dose experimental group have clear structures of the stomach and the intestine, complete mucosal epithelium and no defect, and the pathological changes of congestion, edema, inflammatory cell infiltration and the like are not seen in each layer under the mucosa.

Testis: the development of all levels of spermatogenic cells in the seminiferous tubules is good, interstitial blood vessels do not have congestion and extravasated blood, and interstitial tissues do not have inflammatory cell infiltration and calcific foci.

Ovary: all the follicles are well developed, and interstitial vessels are not congested and extravasated and inflammatory cell infiltration is not seen.

Spleen: the white marrow is composed of a plurality of lymphocytes, the red marrow is composed of medullary cables and blood sinuses, a plurality of red blood cells and a small amount of white blood cells are visible in the blood sinuses, the proportion of the red marrow and the white marrow is not abnormal, and the proportion of the red blood cells and the white blood cells in the blood sinuses is not abnormal.

The examination result shows that: no histopathological changes related to the samples used in the test were observed in the liver, kidney, stomach, intestine, spleen, testis, and ovary of the male and female rats in the control group and the high-dose group.

Clinical examination, hematology examination, blood biochemistry examination and organ weighing result show that the difference is not significant when the experimental group and the control group of each test item are compared. The results of the histopathological examination show that the liver, kidney, stomach, intestine, spleen, testis and ovary of the female rats and the male rats of the control group and the high-dose group have no histopathological changes related to the experimental samples.

14. And (4) testing results:

according to the GB15193-94 standard of food safety toxicology evaluation program and method, the test result is judged as that under the laboratory condition, the sample has oral acute toxicity (L D50) of more than 10.00 g/kg-bw for big and small mice of two sexes, and has no toxic substance according to the acute toxicity grading, the results of three genetic toxicity tests (Ames test, mouse bone multiple erythrocyte micronucleus test and mouse sperm malformation test) are negative, the results of clinical examination, organ weighing, hematology chemical examination and hematology examination in the rat feeding test for 30 days show that the experimental groups of each test item are compared with the control group, and the difference is not significant.

II, auxiliary hypoglycemic action function test:

1. description of the samples:

the sample was a capsule as described in example 1, the contents being a brown powder, 0.4 g/pellet. Sealed and moisture-proof for preservation, and has a shelf life of 24 months.

2. Experimental animals:

the experiment selects healthy second-grade Kunming female mice (license number: SCXK11-00-0006) provided by the experimental animal center of Chinese medical academy of sciences, and the weight of the mice is 26 +/-2 g.

3. Selecting the dosage:

the test object is a setting product with the human body recommended dose of 2.4g/60kg · bw per day, the equivalent dose of the mouse is 10 times of the human body recommended dose, 5 times, 10 times and 30 times of the human body recommended dose are respectively used as low (0.2g/kg · bw), medium (0.4g/kg · bw) and high dose (1.2g/kg · bw), the stomach filling method is adopted, the stomach filling is performed once per day, the stomach filling volume is 0.1m L/10 g of the weight of the mouse, and after the test object is continuously given for 30 days, all indexes are measured.

4. The instrument comprises the following steps:

a spectrophotometer model 755, a thermostatic water bath and a centrifuge.

5. Reagent:

(1) glucose assay kit (glucose oxidase method), manufactured by Beijing Zhongsheng bioengineering high tech company.

(2) Tetraoxypyrimidine, manufactured by SIGMA corporation of America.

6. The detection method comprises the following steps:

(1) effect on blood glucose in normal mice:

mice were fasted for 4 hours, allowed free access to water, and blood was taken from the angular venous plexus to measure fasting blood glucose values, which were divided into two groups according to blood glucose levels: blank control group and test object group. By adopting the intragastric administration method, the test substance group is intragastric administered with high dose of test substance, and the blank control group is intragastric administered with distilled water with corresponding volume. After continuously feeding for 30 days, the fasting blood sugar value is measured, and the influence of the test substance on the blood sugar of normal mice is observed.

(2) Effect on blood glucose in alloxan-induced hyperglycemia model mice:

after 24 hours of fasting, injecting physiological saline solution (40mg/kg · bw) of alloxan into tail vein, on 7 days after injection, fasting for 4 hours, freely drinking water, determining fasting blood sugar value, selecting the animals with blood sugar value of 10.0 mmol/L and 25.0 mol/L as successful animals of hyperglycemia model, and randomly dividing the animals into four groups according to blood sugar level, namely, a hyperglycemia model control group, a low dose group, a middle dose group and a high dose group, and the low, middle and high dose groups are filled with test substances with corresponding dose.

(3) Sugar tolerance test:

the hyperglycemia model animals were fasted for 4 hours, the test substances at different concentrations were administered to each dose group, the hyperglycemia model control group was gavaged with distilled water in a corresponding volume, glucose was orally administered at 2.0g/kg · bw after 20 minutes, and blood glucose values at 0 hour, 0.5 hour, and 2 hours after the administration of glucose were measured.

7. Experimental unit: beijing university institute of culture and literature college of application health food function detection center

8. Experimental data analysis of variance was performed using microsoft excel software.

9. The experiment summary:

(1) effect on blood glucose in normal mice:

after the test substance is administered to the normal mice for 30 days, the blood sugar value of the test substance group has no significant difference compared with that of the blank control group (P is more than 0.05).

(2) Effect on fasting plasma glucose in alloxan hyperglycemic model mice:

before the test object is given, the blood sugar value of each dose group of the hyperglycemia model mouse has no obvious difference (P is more than 0.05) compared with the hyperglycemia model control group. After 30 days of administration of different doses of the test substance, the blood glucose value of each dose group has no significant difference compared with the hyperglycemia model control group (P is more than 0.05).

(3) Effect on glucose tolerance in allo-hyperglycemic model mice by allo-uracil:

blood glucose values of the medium and high dose groups are respectively reduced by 13 percent (P is less than 0.05) and 13 percent (P is less than 0.05) compared with the hyperglycemia model control group 0.5 hour after the intragastric glucose administration. The blood sugar value of the high-dose group and the hyperglycemia model control group are reduced by 23 percent (P is less than 0.05) in comparison 2 hours after the intragastric glucose administration.

(4) Effect on mouse body weight:

① effect on body weight of Normal mice:

before and after the test object is given, the body weight of the test object group of the normal mice has no significant difference compared with that of a blank control group (P is more than 0.05).

② Effect on hyperglycemic model mouse body weight:

before and after the test object is given, the body weight of each dose group of the hyperglycemia model mouse has no obvious difference compared with the hyperglycemia model control group (P is more than 0.05).

10. And (4) testing results:

after the mice are orally given with different doses of samples for 30 days, the fasting blood glucose value of the normal animal test object group has no significant difference compared with that of a blank control group (P is more than 0.05); before and after the test object is given, the body weight of the test object group has no significant difference compared with that of a blank control group (P is more than 0.05). The fasting blood glucose value of each dose group of the hyperglycemia model animal has no significant difference (P is more than 0.05) compared with the hyperglycemia model control group. In a glucose tolerance experiment, the blood sugar values of the medium and high dose groups are respectively reduced by 13 percent (P is less than 0.05) and 13 percent (P is less than 0.05) compared with a hyperglycemia model control group 0.5 hour after the intragastric glucose administration; the blood sugar value of the high-dose group is reduced by 23 percent (P is less than 0.05) compared with the hyperglycemia model control group 2 hours after the intragastric glucose administration. Before and after the test object is given, the weight of each dose group has no significant difference compared with the hyperglycemia model control group (P is more than 0.05).

Judging according to a food safety toxicology evaluation program and a method: the sample has auxiliary blood sugar lowering effect.

And thirdly, assisting in reducing blood sugar, and performing a human body eating test:

1. description of the samples:

the sample was a capsule as described in example 1, the contents being a brown powder, 0.4 g/pellet. Sealing, storing in moisture-proof manner, and keeping shelf life of 24 months; placebo capsules, supplied by Beijing to Yuanshengshi technology development, Inc.

2. Subject selection:

① it is stable after diet control or oral hypoglycemic agent treatment, and only takes maintenance amount of adult type II diabetic patients with fasting blood glucose of 7.8 mmol/L (140mg/d1) or 2h postprandial blood glucose of 11.1 mmol/L (200mg/dl), or selected hyperglycemia population with fasting blood glucose of 7.8 mmol/L or 6.7 mmol/L (120mg/dl) or 11.1 mmol/L (200mg/dl) or 2h postprandial blood glucose of 7.8 mmol/L.

② exclusionary criteria:

a. those under 18 or 65 years of age, pregnant or lactating women.

b. Has serious complications of gravity, liver, kidney, etc. or other serious primary diseases and psychosis.

c. If the result is not good, the curative effect or the data are not complete.

3. The test feeding method comprises the following steps:

60 subjects are randomly divided into two groups, the varieties and the dosages of the original hypoglycemic drugs are unchanged, the hypoglycemic capsules described in the embodiment 1 are added to a sample group, and placebo is added to a control group, wherein the dosage of the hypoglycemic capsules is 3 times a day, and 2 capsules are taken each time. The test taker is required to adhere to diet control for a long time and eat according to different labor intensity and body types. Each group adopts self-contrast design, and two groups adopt inter-group contrast design.

4. Instruments and reagents:

the model F-820 blood-ball counter, MIDIRON urine ten-item analyzer (Demade), RA1000 full-automatic biochemical analyzer model (American made), biochemical test box are all provided by Zhongsheng company.

5. Observation indexes are as follows:

each index is tested once at the beginning and the end of the test feeding experiment, and fasting blood sugar is tested once on the 15 th day of the test feeding.

(1) And (3) observing the efficacy:

① Observation of symptoms:

the medical history is inquired in detail, the diet condition and the activity of the patient are known, and the main clinical symptoms are observed: polyphagia, polydipsia, diuresis, fatigue, weakness, etc. The integral values were counted before and after the test for the mild symptoms (3 points of severe, 2 points of moderate, 1 point of mild symptoms), and the symptom improvement rates were observed for the improvement of the main symptoms (2 points of improvement for each symptom were significant, and 1 point of improvement was significant).

② fasting blood glucose and 2 hours postprandial blood glucose test.

③ urine diabetic ketone body detection.

④ blood lipid (total cholesterol TC, triglyceride TG) detection.

(2) And (4) safety observation:

① routine examination of hematuria:

red blood cell count, hemoglobin, white blood cell count, urine ten measurements.

② biochemical index determination:

serum albumin a L B, total protein TP, heart, liver and kidney functions (aspartate aminotransferase AST, glutamate pyruvate transaminase a L T, UREA, inosine CRE).

③ abdominal B-mode ultrasonography, electrocardiogram, and X-ray chest fluoroscopy.

(3) Efficacy determination criteria:

① is effective in improving basic symptoms and reducing blood sugar by 10% or more compared with that before trial.

② is ineffective, the basic symptoms are not improved obviously, and the blood sugar is reduced to the standard.

6. Experimental unit: beijing disease prevention and control center (Western Yuan Hospital of Chinese institute of medical science)

7. The experiment summary:

(1) general data:

a total of 60 cases were observed, and the group of samples was 15 males and 15 females, with the age of minimum 38 years, the age of maximum 65 years, the mean age of 54.40 years, and the mean course of disease of 6.03 years; control group 15 men and 15 women, the age was 31 years minimum, 65 years maximum, 53.70 years mean, and the course was 5.63 years mean.

(2) Two groups were compared for general conditions before observation:

before the two groups of test foods, the indexes have no obvious difference and are comparable.

(3) The blood sugar reducing effect is as follows:

① clinical observations:

clinical symptoms changed before and after the test feeding, and the control P between groups was less than 0.05.

② comparison of fasting and postprandial blood glucose:

a. comparing fasting blood sugar before and after test eating of the sample group and the control group:

after one month, the fasting blood sugar of the sample group is reduced by 1.09 mml/L, the fasting blood sugar of the control group is not obviously reduced, and the two groups have obvious difference.

b. Comparing the postprandial 2-hour blood sugar of the sample group and the control group before and after test eating:

after one month, the blood sugar of the sample group decreases by 1.20 mmol/L after 2 hours, the self-control difference is obvious, the blood sugar of the control group after 2 hours is not obvious, and the two groups have obvious difference.

(4) Blood lipid change:

the blood lipid changes before and after the test eating are not obviously different.

(5) Blood safety index observation:

before and after the two groups of test meals, all indexes of blood detection are in a normal range.

(6) Chest X-ray, electrocardiogram and B-ultrasonic examination:

the subjects were all in the normal range.

10. And (4) testing results:

by adopting a self-control method and a group-group control method, 60 hyperglycemic test feeders are randomly divided into a sample group and a control group, 30 capsules in each group have unchanged varieties and dosages of hypoglycemic drugs originally used in an observation period, the sample group is added with the capsules described in the example 1, the control group is added with a placebo, after one month, the result shows that the capsules described in the example 1 have the effect of reducing blood sugar, the fasting blood sugar is reduced by 1.09 +/-1.65 mmol/L, the postprandial blood sugar is reduced by 1.20 +/-1.85 mmol/L, 19 effective cases have 19 effective cases, the total effective rate is 63.33%, the fasting blood sugar and postprandial blood sugar in the control group are not obviously reduced, 9 effective cases have the total effective rate of 30.00%, the two groups have obvious difference, the capsules described in the example 1 have the effect of reducing blood sugar in an auxiliary manner, hemoglobin, red blood cells, white cells, serum, total protein, albumin, glutamic-oxaloacetic transaminase, propyltransaminase, urea, glucoside, urine convention and other clinical indexes are in a normal range, and the capsules described in the example 1 have no obvious influence on physical health of the test feeders, and no adverse effect on other symptoms of polyphagia diet and other main process.

According to judgment of 'evaluation procedure and inspection method of health food functionality', the test results of the test food show that the capsule described in example 1 has the auxiliary hypoglycemic effect.

Fourthly, hygiene inspection, functional component inspection and stability inspection:

1. sample preparation: the capsules of example 1, 0.4 g/capsule, were stored at room temperature.

2. The inspection basis is as follows: GB/T5009 food hygiene physical and chemical inspection method, pharmacopoeia 2000 edition, health food effective component detection method, GB4789-1994 food hygiene inspection method, microbiology division, health department's internal system method, etc.

3. The inspection mechanism comprises: shandong university sanitation analysis testing center

4. And (4) testing results:

(1) and (3) hygiene examination: the sample is tested according to the method specified in the national standard, and the result shows that the relevant indexes of the sample hygiene conform to the national standard.

(2) And (4) testing functional components: the sample is subjected to a heat preservation experiment for three months at 37 ℃ and 75% relative humidity, and the quality of the detected index is stable through the functional component project inspection.

(3) And (3) accelerated stability test: the sample is subjected to a heat preservation experiment for three months at 37 ℃ and 75% of relative humidity, and physicochemical and microbial detection shows that the quality of the detected index is stable.

The capsule specification is 0.4 g/capsule, and each 100g of the capsule contains: 1.01g of total soap, 1.79g of total flavone, 9.3g of calcium and 63mg of chromium picolinate. Taken twice a day, 3 granules at a time.

The above description is only for the specific embodiments of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art can easily conceive of the changes or substitutions within the technical scope of the present invention, and all the changes or substitutions should be covered within the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the appended claims.

Claims

1. The propolis balsam pear composition with the auxiliary hypoglycemic effect is characterized by comprising the following raw material components: 1000 portions of propolis, 4000 portions of balsam pear, 6000 portions of balsam pear, 4000 portions of polygonatum odoratum, 6000 portions of chromium source, 1-5 portions of chromium source and 1000 portions of calcium source, 1800 portions.

2. The propolis-bitter gourd composition with the auxiliary hypoglycemic effect according to claim 1, wherein the raw material components are as follows: 1400 parts of propolis, 5000 parts of balsam pear, 5000 parts of fragrant solomonseal rhizome, 3 parts of chromium source and 1400 parts of calcium source.

3. The propolis-momordica charantia composition with the auxiliary hypoglycemic effect according to claim 1 or 2, wherein the chromium source is a trivalent chromium source.

4. The propolis-bitter gourd composition with the auxiliary blood sugar reducing effect according to claim 3, wherein the trivalent chromium source is one or a mixture of more of chromium picolinate, chromium yeast or chromium polynicotinate, and the calcium source is one or a mixture of more of pearl powder, calcium citrate, L-calcium threonate and L-calcium lactate.

5. A method for preparing the propolis balsam pear composition with the auxiliary hypoglycemic effect according to any one of claims 1 to 4, which is characterized by comprising the following steps:

6. The method according to claim 5, wherein in the step (1), the sieving is 80-120 mesh sieving;

7. The method according to claim 5, wherein in the step (2), the dipping extraction is repeated three times, and the mass of each time of adding ethanol is 3-5 times of the mass of the propolis fine powder;

8. The method according to claim 5, wherein in the step (3), the relative density of the propolis extract is 1.35 to 1.40.

9. The method according to claim 5, wherein in the step (5), the reflux extraction is repeated three times, and ethanol is extracted from each reflux extraction with a volume concentration of 60-75%;

the amount of ethanol added in the first reflux extraction is 8 times of the mass of the mixed material, and the time of the first reflux extraction is 2 hours; the amount of ethanol added in the second reflux extraction is 6 times of the mass of the mixed material, and the time of the second reflux extraction is 1.5 hours; the amount of ethanol added in the third reflux extraction is 4 times of the mass of the mixed material, and the time of the third reflux extraction is 1 h;

in the step (6), the relative density of the extract is 1.15-1.20;

in the step (7), the granularity of the extract powder is 70-90 meshes.

10. The method according to claim 5, wherein in the step (9), the volume concentration of the ethanol is 88 to 92%, and the particle size after the granulation is 18 to 22 mesh.