CN115501293A - Preparation method of prepared rehmannia root, chinese yam and kudzuvine root hypoglycemic composition - Google Patents
Preparation method of prepared rehmannia root, chinese yam and kudzuvine root hypoglycemic composition Download PDFInfo
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- CN115501293A CN115501293A CN202211085463.3A CN202211085463A CN115501293A CN 115501293 A CN115501293 A CN 115501293A CN 202211085463 A CN202211085463 A CN 202211085463A CN 115501293 A CN115501293 A CN 115501293A
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- China
- Prior art keywords
- extraction
- preparation
- water
- rehmanniae preparata
- chinese yam
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Links
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Classifications
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Abstract
The invention discloses a preparation method of a hypoglycemic composition containing radix rehmanniae preparata, chinese yam and kudzu vine root, belonging to the field of extraction of plant active substances. The method comprises the following steps: mixing radix rehmanniae Preparata, corni fructus, rhizoma Dioscoreae, poria and radix Puerariae, grinding, ultrasonic extracting with water, precipitating with ethanol, and supercritical extracting to obtain extract, and mixing the extracts to obtain the composition. The invention takes the rehmanniae praeparatum glycoside D in the prepared rehmannia as a representative component, and keeps different types of active effective components such as polysaccharide, volatile component, glycoside and the like as far as possible through multiple extractions. According to the invention, a proper amount of acidic antioxidant is added in the grinding step, so that the easily oxidized substances can be prevented from deteriorating and losing functions in the extraction process; and the added antioxidant belongs to an edible additive, and is non-toxic and harmless. The test proves that the sugar-controlling composition extracted by the invention has statistically significant control effect on blood sugar.
Description
Technical Field
The invention belongs to extraction of plant active substances, and particularly relates to a preparation method of a hypoglycemic composition containing prepared rehmannia root, chinese yam and kudzu vine root.
Background
The glucose in the blood is called blood sugar (Glu), which is a necessary guarantee for the normal activities of the human body; the normal blood sugar value of a normal person is 3.9-6.1mmol/L when the person has an empty stomach, and the blood sugar value has guiding significance for treating diseases and observing the diseases; fasting plasma glucose of more than 7.0mmol/L is likely to be diabetic.
Diabetes is a chronic disease, is a metabolic disorder of protein, fat and carbohydrate caused by insufficient insulin in vivo, and is mainly characterized by chronic hyperglycemia. Many natural antidiabetic active ingredients have been found, such as: ginkgo leaf extract, plant polysaccharide, etc.
At present, aiming at diabetes, the quality of medicines such as externally injected insulin, dimethyldiconium, sulfonylurea and the like is generally used; however, diabetes is a disease which cannot be cured once at present, and generally, the single treatment effect is not ideal, so that a plurality of different treatment methods are required to jointly treat the diabetes to achieve a better control effect.
The bi sheng yuan brand radix rehmanniae preparata, chinese yam and radix puerariae granules (guo shi jian zhu G20070170) are health food, which takes radix rehmanniae preparata, dogwood, chinese yam, tuckahoe, radix puerariae, tea polyphenol and chromium picolinate as raw materials; the production process comprises the steps of adding water, decocting, extracting, concentrating, spray drying and granulating; has certain auxiliary blood sugar reducing and health care functions. However, the product has been produced for more than ten years, and the addition of chromium picolinate has certain psychological barrier for some consumers with purer plant sources; both the formula and the production process have certain room for improvement.
Disclosure of Invention
The invention provides a preparation method of a hypoglycemic composition containing prepared rehmannia root, chinese yam and kudzu root, which is improved based on Bishengyuan brand prepared rehmannia root, chinese yam and kudzu root granules; the hypoglycemic effect is achieved in a non-medicinal way by mainly extracting components such as polysaccharide, volatile matters, glycosides and the like in the raw materials.
In order to achieve the purpose, the invention adopts the following technical scheme:
the preparation method of the hypoglycemic composition containing the prepared rehmannia root, the Chinese yam and the kudzuvine root is characterized by comprising the following steps of:
(1) Mixing radix rehmanniae Preparata, corni fructus, rhizoma Dioscoreae, poria and radix Puerariae, and grinding to obtain raw materials;
(2) Performing microwave ultrasonic water extraction on the raw material group, and performing solid-liquid separation to obtain a solid phase substance and a water extraction solution;
(3) Precipitating the water extract with ethanol, and performing solid-liquid separation to obtain ethanol precipitate;
(4) Solid phase substance CO 2 Performing supercritical extraction, collecting extract, removing solvent, and collecting extract;
(5) Mixing the ethanol precipitate and the extractive solution, vacuum concentrating, and spray drying;
in the step (1), citric acid and vitamin C are added, and the adding amount is 0.15-0.25% of the total mass of the raw material group; the mass ratio of the citric acid to the vitamin C is 1;
in the step (4), an entrainer is added for extraction; the entrainer is ethanol; the flow rate of the entrainer is 2.0-3.0mL/min; CO 2 2 The fluid pressure is 160-220bar; CO 2 2 The flow rate is 3.5-4.5L/min; the extraction temperature is 30-45 ℃; the extraction time is 3-4h.
In particular, in the step (1), the mixture is ground to be more than or equal to 40 meshes.
Particularly, in the step (2), the ultrasonic power is 300-450W, the intermittent ratio is 4S/1S, and the microwave power is 400-550W.
Particularly, in the step (2), the water extraction temperature is 35-60 ℃, the water extraction time is 40-60min, and the addition amount of water is 12-15 times of the mass of the raw material group.
Particularly, in the step (3), the temperature of alcohol precipitation is 0-25 ℃, and the volume fraction of ethanol in the alcohol precipitation system is 70-85%; the alcohol precipitation time is 6-8h.
In particular, in the step (5), spray drying is carried out until the moisture is less than or equal to 4%.
Adding a proper amount of acidic antioxidant in the step (1) to prevent the easily oxidized substances from deteriorating and losing functions in the extraction process; the added antioxidant belongs to edible additives, can gradually volatilize in the extraction process, and is non-toxic and harmless even if residual exists.
The microwave ultrasonic extraction of step (2) can dissolve the water-soluble substances as much as possible, and the invention finds that the acidic antioxidant can also contribute to the process.
The main purpose of step (3) is to extract components such as polysaccharide and digitoxin D.
The purpose of step (4) is to retain the active ingredients of the raw materials as much as possible, and in this case, the substances that are not easily dissolved in an acidic environment are also extracted in the above process.
Compared with the prior art, the invention has the following beneficial effects:
(1) The invention takes the rehmanniae praeparatum glycoside D in the prepared rehmannia root as a representative, and keeps different types of active effective components such as polysaccharide, glycoside and the like as far as possible through multiple extractions.
(2) According to the invention, a proper amount of acidic antioxidant is added in the grinding step, so that the easily oxidized substances can be prevented from deteriorating and losing functions in the extraction process; and the added antioxidant belongs to an edible additive, and is non-toxic and harmless.
(3) Tests prove that the sugar-controlling composition extracted by the invention has a statistically significant control effect on blood sugar.
Detailed Description
For a better understanding of the present invention, the present invention is further described in conjunction with the following specific examples, wherein the terminology used in the examples is for the purpose of describing particular embodiments only and is not intended to limit the scope of the present invention.
In the invention:
the radix rehmanniae Preparata is processed product of fresh or dried root tuber of radix rehmanniae Preparata Rehmannia glutinosa Libosch of Scrophulariaceae. The method for treating rehmanniae radix Preparata is described in pharmacopoeia.
The Corni fructus is dried mature pulp (core removed) of Cornus officinalis Sieb.
The rhizoma Dioscoreae is dried rhizome of Dioscorea opposita oppysita Thunb of Dioscoreaceae.
Poria is dried sclerotium of Wolf of Poria cocos (Schw.) Cos (belonging to Polyporaceae).
Radix Puerariae is dried root of Ohwi of Pueraria lobata (Willd.) Ohwi of Leguminosae.
Example measurement of raw Material content
The content of digitoxin D in prepared rehmannia root is determined according to the record of pharmacopoeia 2020 edition.
The remaining ingredients in rehmanniae radix Preparata, such as catalpol, leonuroside, verbascoside and isoverbascoside, were determined according to the description in comparison of the content of 5 glycosides and total polysaccharides in rehmanniae radix and rehmanniae radix Preparata (Zhang-ken, kinsenosia, tuwanqian, etc., natural products research and development, nat Prod Res Dev 2019, 31.
The morroniside and loganin content of Corni fructus is determined according to the record of pharmacopeia 2020 edition.
The content of rhizoma Dioscoreae polysaccharide in rhizoma Dioscoreae is determined according to the record of "extraction and separation of rhizoma Dioscoreae polysaccharide and content determination of rhizoma Dioscoreae total polysaccharide" (Wanggang, dushiming, shaoshengsheng, etc., china Hosp Pharm J,2007Oxt, vol 27.No.10, volume 27, 2007 in pharmaceutical journal of China hospital).
The content of polysaccharides in Poria cocos is determined according to the records in "content determination of polysaccharides in Poria peel and Poria cocos blocks Li" (Junyan, wei scholar, shichaoshan et al, qian nan national medical science report 2009, vol.22, no. 4).
The content of puerarin in radix Puerariae is determined according to record of pharmacopoeia 2020 edition.
The plant materials used in the present invention were measured for the content of active ingredients, and the results are shown in table 1 below, and the content is all in%.
TABLE 1
Example two extraction Rate experiments
The experimental preparation was prepared as follows.
(1) Mixing radix rehmanniae Preparata, corni fructus, rhizoma Dioscoreae, poria and radix Puerariae, adding citric acid and vitamin C, and grinding to more than 40 mesh to obtain raw material group;
the amounts of citric acid and vitamin C added are shown in table 2 below;
(2) Performing microwave ultrasonic water extraction on the raw material group, and performing solid-liquid separation to obtain a solid phase substance and a water extraction solution;
the intermittent ratio in the step (2) is 4S/1S; the ultrasonic power, the microwave power, the water extraction temperature, the water extraction time and the added amount of water are shown in table 2;
(3) Precipitating the water extract with ethanol, and performing solid-liquid separation to obtain ethanol precipitate;
in the step (3), the alcohol precipitation temperature, the volume fraction of ethanol in the alcohol precipitation system and the alcohol precipitation time are shown in table 2;
(4) Adding entrainer into solid phase for CO treatment 2 Supercritical extracting, collectingCollecting the extraction solution to remove solvent, and taking the extraction solution;
in the step (4), the entrainer is ethanol; entrainer flow rate, CO 2 Fluid pressure, CO 2 The flow rate, extraction temperature, and extraction time are shown in table 2;
(5) Mixing the alcohol precipitate and the extractive solution, vacuum concentrating, and spray drying to water content of 4% or less.
TABLE 2
Respectively taking raw materials before preparation for finished products with the serial numbers of I, II, III and IV in the embodiment according to the method in the embodiment; the results of comparing the extracted products obtained by separately extracting different materials from each serial number in the same manner are shown in table 3 below.
More specifically, the same method for extracting the product from different raw materials in each serial number separately means that the radix rehmanniae preparata, the cornus officinalis, the yam, the poria cocos, and the pueraria lobata are extracted respectively according to the parameters from the step (1) to the step (4) in the above table 2.
Meanwhile, for the sake of comparison, the content of the raw materials corresponding to the serial numbers is the same as the amount of all the raw materials in the previous step, but not the same as the raw material amount of the product. If the raw material powder of rehmanniae radix preparata is weighed as 1g, the prepared rehmanniae radix preparata raw material is also used in an amount of 1g.
In summary, the whole comparison process takes sequence number one as an example:
1. different calibration curves were prepared by weighing 1g of the starting powder.
2. Weighing radix rehmanniae Preparata, corni fructus, rhizoma Dioscoreae, poria and radix Puerariae corresponding to 1g.
3. Preparing 5 corresponding finished products which independently contain the unique plant raw materials by using different step parameters in the sequence number I; the amount of the group of material at this point is the mass of the corresponding unique plant material.
4. The product was compared with other active ingredients in the same manner as in example 1.
TABLE 3
The catalpol content is too low, so the extraction rate is not applicable.
Example three animal experiments
The compositions prepared in the above examples were used for animal experiments.
1) Materials and methods
The test substance: the compositions in table 2.
Animals: 150 female mice of Kunming breed of clean grade, weight 26 + -2 g, provided by laboratory animal farm of animal science and technology institute of Hunan university of agriculture. The room temperature is 23-24 ℃, and the relative humidity is 56-58%.
2) The test method comprises the following steps:
the mice were randomly divided into 15 groups by weight, 10 mice per group, 130 mice were modeled for hyperglycemia model, and 20 mice were used as normal animals.
Setting three test object dose groups (low, medium and high doses) according to the recommended dose of a human body, wherein the dose groups are respectively 0.01 g/kg-bw, 0.02 g/kg-bw and 0.06 g/kg-bw; the test substance is added with distilled water to 200mL to prepare corresponding dosage concentration. The gavage is carried out once a day, the gavage volume is 0.2mL/10g · bw, and the control group is given distilled water with the same volume once a day for 30 consecutive days.
2.1 Molding
150 mice are selected to be fasted for 24 hours, then alloxan (40 mg/kg. Bw) is injected into tail vein, after 5 days, the mice are fasted for 5 hours, and then blood sugar value is measured, and the mice with the blood sugar value of 10-25mmol/L are successful animals of a hyperglycemia model. Randomly dividing the model into a model control group and 12 dose groups after the model selection is successful; the test substances with different concentrations are given to the dosage group, the distilled water with the same volume is given to the model group, the fasting blood sugar value is measured when fasting is performed for 5 hours for 30 days continuously, and the blood sugar value and the blood sugar reduction percentage of each component animal are compared.
Percent blood glucose decrease = (blood glucose before experiment-blood glucose after experiment)/blood glucose before experiment × 100%.
2.2 Normal animals
20 mice were divided into 1 control group and 1 test group at random in accordance with 5-hour fasting blood glucose level, the test group was given a test substance of high dose concentration, the model group was given distilled water of the same volume for 30 consecutive days, and fasting blood glucose level was measured at 5-hour fasting state, and blood glucose level and percent blood glucose decrease of each component animal were compared.
Percentage of blood glucose decrease = (blood glucose value before experiment-blood glucose value after experiment)/blood glucose value before experiment × 100%.
2.3 sugar tolerance test
Hyperglycemia model animals were fasted for 5 hours, test substances of different concentrations were administered to the dose groups, the same volume of distilled water was administered to the model control group, glucose was orally administered for 15 to 20 minutes at 2.0g/kg · bw, blood glucose levels at 0, 0.5, and 2 hours after glucose administration were measured, and changes in area under the blood glucose curve at each time point after glucose administration to the model control group and the test group were observed.
Area under blood glucose curve =0.25 × (blood glucose level at 0 hour + blood glucose level at 4 × 0.5 hour + blood glucose level at 3 × 2 hours)
3) As a result, the
3.1 body weight
As shown in tables 4-1 and 4-2, the initial body weight, the intermediate body weight and the final body weight of the mice are not obviously different from those of the control group when the high dose test object is orally administered to the mice in different examples (P > 0.05).
TABLE 4-1 Normal mouse weights
| Group of | Animal number (only) | Initial body weight (g) of experiment | Middle-term body weight of experiment (g) | End of test body weight (g) |
| Control group | 10 | 25.87±1.23 | 33.88±2.06 | 39.05±1.85 |
| Number one high dose group | 10 | 25.73±1.46 | 33.42±1.38 | 39.15±2.29 |
TABLE 4-2 hyperglycemic mouse body weights
| Group of | Animal number (only) | Initial body weight (g) of experiment | Middle-term body weight (g) of experiment | End of test body weight (g) |
| Control group | 10 | 25.81±1.37 | 23.13±1.58 | 25.21±1.39 |
| Number one high dose group | 10 | 26.14±1.14 | 25.76±1.05 | 25.38±1.25 |
| Serial number two high dose group | 10 | 25.91±1.17 | 26.21±1.61 | 26.24±1.46 |
| Serial number three high dose group | 10 | 25.42±1.21 | 26.34±1.57 | 25.84±1.54 |
| Serial number four high dose group | 10 | 26.07±1.42 | 26.36±1.20 | 26.15±1.50 |
3.2 fasting plasma glucose Effect on Normal mice
In Table 5, the high dose of the test substance orally administered to the mice had no significant effect on the blood glucose level and the percentage of blood glucose decrease in normal animals (P > 0.05).
TABLE 5 fasting plasma glucose Effect in Normal mice
| Group of | Animal number (only) | Blood glucose before experiment (mmol/L) | Post-test blood glucose (mmol/L) | Percentage reduction in blood glucose (%) |
| Negative control group | 10 | 4.24±0.72 | 4.45±0.60 | -6.31±14.46 |
| Example a high dose group | 10 | 4.32±0.74 | 4.12±0.59 | 3.73±10.25 |
3.3 Effect on blood glucose in hyperglycemic model animals.
In table 6, when the mice were orally administered with different doses of the test substance, the percentage of blood glucose decrease in the dose groups was increased, and the difference between the medium and high dose groups was significant (P < 0.05).
TABLE 6 Effect on glucose tolerance in hyperglycemic model animals
The above detailed description is specific to one possible embodiment of the present invention, and the embodiment is not intended to limit the scope of the present invention, and all equivalent implementations or modifications without departing from the scope of the present invention should be included in the technical scope of the present invention.
Claims (5)
1. The preparation method of the hypoglycemic composition containing the prepared rehmannia root, the Chinese yam and the kudzuvine root is characterized by comprising the following steps of:
(1) Mixing radix rehmanniae Preparata, corni fructus, rhizoma Dioscoreae, poria and radix Puerariae, and grinding to obtain raw materials;
(2) Performing microwave ultrasonic water extraction on the raw material group, and performing solid-liquid separation to obtain a solid phase substance and a water extraction solution;
(3) Precipitating the water extract with ethanol, and performing solid-liquid separation to obtain ethanol precipitate;
(4) Solid phase substance CO 2 Performing supercritical extraction, collecting extract, removing solvent, and collecting extract;
(5) Mixing the water extractive solution, ethanol precipitate, and extractive solution, vacuum concentrating, and spray drying;
in the step (1), citric acid and vitamin C are added, and the adding amount is 0.15-0.25% of the total mass of the raw material group; the mass ratio of the citric acid to the vitamin C is 1.5-1.2;
in the step (4), an entrainer is added for extraction; the entrainer is ethanol; the flow rate of the entrainer is 2.0-3.0mL/min; CO 2 2 The fluid pressure is 160-220bar; CO 2 2 The flow rate is 3.5-4.5L/min; the extraction temperature is 30-45 ℃; the extraction time is 3-4h.
2. The preparation method of the hypoglycemic composition containing radix rehmanniae preparata, chinese yam and radix puerariae as claimed in claim 1, wherein in the step (2), the ultrasonic power is 300-450W, the intermittent ratio is 4S/1S, and the microwave power is 400-550W.
3. The preparation method of the hypoglycemic composition containing the radix rehmanniae preparata, the Chinese yam and the radix puerariae according to claim 1, wherein in the step (2), the water extraction temperature is 35-60 ℃, the water extraction time is 40-60min, and the addition amount of water is 12-15 times of the mass of the raw material group.
4. The preparation method of the hypoglycemic composition containing the radix rehmanniae preparata, the yam and the radix puerariae according to claim 1, wherein in the step (3), the temperature of alcohol precipitation is 0-25 ℃, and the volume fraction of ethanol in the alcohol precipitation system is 70-85%; the alcohol precipitation time is 6-8h.
5. The preparation method of the hypoglycemic composition containing radix rehmanniae preparata, chinese yam and radix puerariae as claimed in claim 1, wherein in the step (5), the hypoglycemic composition is spray-dried until the water content is less than or equal to 4%.
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