CN113082085A - Cistanche tubulosa extract and preparation method thereof - Google Patents
Cistanche tubulosa extract and preparation method thereof Download PDFInfo
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/64—Orobanchaceae (Broom-rape family)
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- A61K2236/50—Methods involving additional extraction steps
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Abstract
The invention relates to a cistanche tubulosa extract and a preparation method thereof, wherein the cistanche tubulosa extract comprises 25-35% of polysaccharide, 10-15% of total flavone and 7-12% of total glycoside of cistanche tubulosa, and the yield of the extract is more than 10%. The zebra fish drug effect experiment confirms that the extract can effectively promote intestinal peristalsis, and can be developed into a drug or a health-care product for preventing or treating senile constipation and habitual constipation. The method has the advantages of simple and convenient process, economy, environmental protection and suitability for large-scale production.
Description
Technical Field
The invention relates to the field of plant extraction, in particular to a cistanche tubulosa extract with the effect of relaxing bowel and a preparation method thereof. The application is a divisional application of Chinese patent application No. 201610328836.3, which has an application date of 2016, 05 and 17, and has the name of 'cistanche tubulosa extract and a preparation method thereof'.
Background
Along with the change of the dietary structure of people and the influence of psychopsychological and social factors, the incidence rate of constipation tends to increase, and people are puzzled more and more. According to statistics, the prevalence rate of constipation in the population is as high as 27%, and the tendency is high. In addition to the discomfort, constipation patients may also have psychological disorders such as insomnia, anxiety, depression, and the like, and severe patients may have symptoms such as abdominal pain, anemia, and emaciation. The constipation has high morbidity and complex etiology, so the patients often have a lot of troubles, and the life quality is influenced when the constipation is serious. Therefore, the preparation of a medicament for preventing or treating constipation-related diseases with a significant effect has become one of the hot spots in the field of research on medicaments or health-care foods
Cistanche tubulosa is a dry fleshy stem with phosphorus leaves of Cistanche tubulosa (Schenk) r.bright, a parasitic plant of the family orobanaceae, is commonly called as herba cistanches, is sweet in taste, salty and warm in nature, is produced in a tali basin in Xinjiang, is rich in resources, and is listed as the superior product in Shen Ben Cao Jing. The phenylethanoid glycosides are the main active ingredients of cistanche tubulosa, and contain a small amount of iridoid and lignin. Cistanche tubulosa has been used as a Chinese herbal medicine for thousands of years, is particularly widely applied as a kidney-tonifying and yang-strengthening medicine, and has the frequency of being second to that of ginseng in the traditional nourishing formula, namely desert ginseng. Modern pharmacological research shows that cistanche tubulosa also has the effects of enhancing the immune function, resisting fatigue, aging, atherosclerosis, oxidation and virus, protecting the liver, promoting metabolism and the like. Relevant researches show that the cistanche tubulosa has the effect of relaxing bowel and can be used for preventing or treating constipation and other related diseases. The preparation method of the cistanche tubulosa extract for preventing or treating senile constipation and habitual constipation at present is particularly lack. In order to solve the problem, the patent provides a preparation process of the cistanche tubulosa extract, and the effect of relaxing bowel is confirmed through a drug effect experiment. The method has the advantages of simple operation, high yield, low cost, stable product quality, green and environment-friendly process and suitability for large-scale production.
Disclosure of Invention
Therefore, the invention aims to provide an extract of fresh or dry cistanche tubulosa;
the technical scheme for solving the technical problem is as follows:
the cistanche tubulosa extract comprises 25-35% of polysaccharide, 9-12% of oligosaccharide, 10-15% of total flavone, 7-12% of total glycosides of cistanche tubulosa, 5-8% of polyphenol, 5-8% of protein and the balance of water and ash, and the mass of the cistanche tubulosa extract is taken as a reference.
Preferably, the extract comprises 31.35-35% of polysaccharide, 11.56-12% of oligosaccharide, 13.25-15% of total flavone, 11.12-12% of total glycosides of cistanche deserticola, 6.77-8% of polyphenol, 6.94-8% of protein, and the balance of water and ash.
Preferably, the extract comprises 29.44-35% of polysaccharide, 10.83-12% of oligosaccharide, 12.31-15% of total flavone, 11.03-12% of total glycosides of cistanche deserticola, 7.13-8% of polyphenol, 7.61-8% of protein, and the balance of water and ash.
Preferably, the extract comprises 32.22-35% of polysaccharide, 11.79-12% of oligosaccharide, 13.04-15% of total flavone, 11.90-12% of total glycosides of cistanche deserticola, 5.80-8% of polyphenol, 7.06-8% of protein, and the balance of water and ash.
Preferably, the extract comprises 31.3% polysaccharide, 11.56% oligosaccharide, 13.25% total flavone, 11.12% total glycosides of cistanche deserticola, 6.77% polyphenol, 6.94% protein, and the balance water and ash.
Preferably, the extract comprises 29.44% polysaccharide, 10.83% oligosaccharide, 12.31% total flavone, 11.03% total glycosides of cistanche deserticola, 7.13% polyphenol, 7.61% protein, and the balance of moisture and ash.
Preferably, the extract comprises 32.22% polysaccharide, 11.79% oligosaccharide, 13.04% total flavone, 11.90% total glycosides of cistanche deserticola, 5.80% polyphenol, 7.06% protein, and the balance of moisture and ash.
The other purpose of the invention is to provide a method for preparing the cistanche tubulosa extract, which comprises the following steps:
(1) refluxing and extracting fresh or dry cistanche tubulosa with purified water for 2-3 times, 2-5 hours each time; mixing the extracting solutions, filtering and concentrating to a temperature of 40-50 ℃ until the density is 1.0-1.2 g/mL (or the solid content is more than 30 wt%); the purified water is preferably deionized water.
(2) Adding alcohol into the concentrated solution obtained in the step (1), and obtaining a precipitate which is the cistanche tubulosa extract.
Preferably, (1) taking the cistanche tubulosa, slicing, and performing reflux extraction for 2-3 times by using water with the mass being 5-10 times of that of the cistanche tubulosa, wherein each time lasts for 2-5 hours; combining the water extracting solutions, carrying out centrifugal filtration, concentrating the filtrate until the density is 1.0-1.2 g/mL (or the solid content is more than 30 wt%) at 40-50 ℃, using the recovered water for next extraction, and cooling the concentrated solution to 30-50 ℃ for later use;
(2) stirring the concentrated solution in the step (1), adding alcohol to adjust the volume concentration of the alcohol to be 60-70%, standing overnight, centrifuging, removing supernatant, and drying precipitates to obtain the cistanche tubulosa extract.
Preferably, the fresh or dry product of cistanche tubulosa is the above-ground or below-ground part thereof;
preferably, the alcohol may be one or more of methanol, ethanol, isopropanol or n-butanol, more preferably edible or pharmaceutical grade ethanol is used;
more preferably, the drying mode can be one or more of vacuum drying, normal pressure drying, freeze drying and spray drying.
The invention also provides a cistanche tubulosa extract which is characterized by being prepared by the method. Preferably, the extract prepared by the method comprises 25-35% of polysaccharide, 9-12% of oligosaccharide, 10-15% of total flavone, 7-12% of total glycosides of cistanche deserticola, 5-8% of polyphenol, 5-8% of protein, and the balance of water and ash, wherein the mass of the cistanche tubulosa extract is taken as a reference. More preferably, the method can also prepare the more preferred extract containing polysaccharide, oligosaccharide, total flavone, total glycosides of cistanche deserticola, polyphenol, protein, and the balance of moisture and ash.
Another object of the present invention is to provide a pharmaceutical composition comprising the extract of the present invention:
a pharmaceutical composition comprises the above cistanche tubulosa extract and edible or medicinal adjuvants.
Preferably, the pharmaceutical composition is tablets, dispersible tablets, capsules, soft capsules, microcapsules, granules, injections, powder injections, freeze-dried powder injections, micro-pills, dripping pills, syrups, powders, extracts, soft extracts, oral liquid preparations and other pharmaceutically acceptable dosage forms.
The invention also aims to provide the application of the cistanche tubulosa extract in preparing medicines or health-care products for increasing intestinal peristalsis and preventing or treating senile constipation and habitual constipation.
Specifically, the technical problem of the invention is realized by the following method:
taking fresh or dry cistanche tubulosa, slicing, extracting with water for multiple times, precipitating with ethanol, centrifuging, removing supernatant, and drying precipitates to obtain the cistanche tubulosa extract with the yield of more than 10%, wherein the extract comprises 25-35% of polysaccharide, 9-12% of oligosaccharide, 10-15% of total flavonoids, 7-12% of total glycosides of cistanche tubulosa, 5-8% of polyphenol, 5-8% of protein, and the balance of water and ash, and the mass of the cistanche tubulosa extract is taken as a reference. The zebra fish drug effect experiment confirms that the extract can effectively promote intestinal peristalsis, and can be developed into a drug or a health-care product for preventing or treating senile constipation and habitual constipation. The method has the advantages of simple and convenient process, economy, environmental protection and suitability for large-scale production.
The specific technical scheme is as follows:
(1) taking cistanche tubulosa, slicing, and performing reflux extraction for 2-3 times by using water with the mass being 5-10 times of that of the cistanche tubulosa, wherein each time lasts for 2-5 hours;
(2) combining the water extracting solutions obtained in the step (1), carrying out centrifugal filtration, concentrating the filtrate until the density is 1.0-1.2 g/mL (or the solid content is more than 30 wt%) when the temperature is 40-50 ℃, using the recovered water for next extraction, and cooling the concentrated solution to 30-50 ℃ for later use;
(3) and (3) stirring the concentrated solution in the step (2), adding alcohol to adjust the volume concentration of the alcohol to be 60-70%, standing overnight, centrifuging, removing supernatant, and drying precipitates to obtain the cistanche tubulosa extract.
The cistanche tubulosa can be fresh or dry products, and can also be a mixture of the fresh and dry products.
The alcohol may be one or more of methanol, ethanol, isopropanol or n-butanol, preferably edible or pharmaceutical grade ethanol is used.
The drying mode can be one or more of vacuum drying, normal pressure drying, freeze drying and spray drying.
The cistanche tubulosa extract prepared in the patent can be compounded and combined with conventional medicaments for preventing or treating constipation, such as fructus cannabis, bitter apricot seeds and the like.
The cistanche tubulosa extract prepared in the patent can be combined with pharmaceutical excipients, and is tablets, dispersible tablets, capsules, soft capsules, microcapsules, granules, injections, powder injections, freeze-dried powder injections, micro-pills, dropping pills, syrups, powder, extracts, soft extracts, oral liquid preparations and other pharmaceutically acceptable dosage forms.
The cistanche tubulosa extract or the pharmaceutical composition thereof prepared in the patent can increase intestinal peristalsis, and is used in the aspect of medicines or health care products for preventing or treating senile constipation and habitual constipation.
Research shows that the cistanche tubulosa extract can obviously increase the intestinal tract peristalsis of zebra fish. Therefore, it is expected that the cistanche tubulosa extract or the pharmaceutical composition containing the same of the present invention can be used in the field of drugs or health products for preventing or treating senile constipation and habitual constipation.
Drawings
Fig. 1 shows quantitative evaluation of zebrafish intestinal fluorescence signals (n-15) in the bioactivity assay of extract G-2. (A) Original data of zebra fish intestinal fluorescence signals; (B) promotion rate of intestinal peristalsis of zebra fish;
FIG. 2 is a graph showing the fluorescence intensity of zebra fish intestinal tracts in the bioactivity test of the extract G-2.
Examples
The invention is further illustrated by the following examples. It should be understood that the method described in the examples is only for illustrating the present invention and not for limiting the present invention, and that simple modifications of the preparation method of the present invention based on the concept of the present invention are within the scope of the claimed invention. All materials and solvents used in the examples were obtained from Sigma Biochemical and Organic Compounds for Research and Diagnostic Reagents, unless otherwise specified.
The first embodiment is as follows:
cleaning cistanche tubulosa (100 kg) (purchased from Xinjiang and Tianli sand, drug development, Ltd.) with tap water, soaking for 60min, slicing (thickness is about 5-8 mm), adding 10 times of water by mass, refluxing and extracting for 3 h, extracting for three times, combining the extracting solutions, centrifuging and filtering, concentrating the filtrate to 50 ℃ density of 1.05G/mL by a single-effect concentrator, cooling to 40 ℃, adding edible ethanol with volume concentration of 95% while stirring to adjust the volume concentration of the alcohol to 70%, standing overnight, centrifuging, removing supernatant, and vacuum drying the precipitate to obtain the cistanche tubulosa extract, wherein the extract contains 31.35% of polysaccharide, 11.56% of oligosaccharide, 13.25% of total flavone, 11.12% of total glycosides of cistanche tubulosa, 6.77% of polyphenol, 6.94% of protein, and the balance of water and ash, and has an extraction rate of 14.8% and a batch number of G-1.
Example two:
cleaning cistanche tubulosa (100 kg) (purchased from Xinjiang and Tianli sand, drug development, Ltd.) with tap water, soaking for 60min, slicing (thickness is about 5-8 mm), adding 8 times of water by mass, refluxing and extracting for 2 h, extracting twice, combining the extracting solutions, centrifuging and filtering, concentrating the filtrate to 50 ℃ density of 1.10G/mL by a single-effect concentrator, cooling to 40 ℃, adding 95% edible ethanol by volume concentration while stirring to adjust the alcohol volume concentration to 65%, standing overnight, centrifuging, removing supernatant, spraying the precipitate to obtain the cistanche tubulosa (drug) extract, wherein the extract contains 29.44% of polysaccharide, 10.83% of oligosaccharide, 12.31% of total flavone, 11.03% of total glycosides of cistanche tubulosa, 7.13% of polyphenol, 7.61% of protein, and the balance of water and ash, the extraction rate is 11.2%, and the batch number is G-2.
Example three:
cleaning cistanche tubulosa (100 kg) (purchased from Xinjiang and Tianli sand drug development, Ltd.) with tap water, soaking for 60min, slicing (thickness is about 5-8 mm), adding 6 times of water by mass, refluxing and extracting for 5 h, extracting twice, combining the extracting solutions, centrifuging and filtering, concentrating the filtrate by a single-effect concentrator until the solid content is 36.2 wt%, cooling to 40 ℃, adding 82% edible ethanol by volume concentration while stirring to adjust the volume concentration of the alcohol to be 62%, standing overnight, centrifuging, removing supernatant, freeze-drying the precipitate to obtain the cistanche tubulosa (cistanche tubulosa) extract, wherein the extract contains 32.22% of polysaccharide, 11.79% of oligosaccharide, 13.04% of total flavone, 11.90% of total glycosides of cistanche, 5.80% of polyphenol, 7.06% of protein, and the balance of water and ash, the extraction rate is 14.1%, and the lot number is G-3.
Example four: evaluation of effect of cistanche tubulosa extract or composition containing cistanche tubulosa extract on intestinal motility
Experimental animals: and AB-line zebra fish is bred in a natural pairing mating mode. Each experimental group had 30 tails and the age was 3 days after fertilization.
The experimental method comprises the following steps: according to the method reported by Zhou et al (Zhou J, Guo SY, Zhang Y, Li CQ. human prokinetic drugs molar mobility in zebrafish. neuroastroreol Motil.2014,26:589-95), zebrafish in a certain growth stage are selected to be fed with Nile Red, and an experimental model for evaluating the effect of the drug on the intestinal peristalsis is constructed: the concentrations of cistanche tubulosa extract batch numbers G-2 are set as follows: 200 mu g/mL, 667 mu g/mL and 2000 mu g/mL, and a model group (experimental model group without drug) and a positive control group (domperidone, 2000 mu g/mL) are arranged at the same time, and 30 tails of the extract, the model group and the positive control group at each concentration are treated for 6 h. After the treatment is finished, photographing is carried out under a fluorescence microscope, the number of randomly photographed images in each group is 15, Total fluorescence signals (S) of intestinal tracts of the zebra fish are analyzed by NIS-Elements D3.10 advanced image processing software, and the intestinal motility promoting rate is calculated according to the following formula: (1-S)Medicine/SModel (model)) X is 100%; the experimental data are expressed as Mean ± SE.
TABLE 1 promotion of Zebra fish intestinal peristalsis by cistanche tubulosa extract (%)
| Sample (I) | 200μg/mL | 667μg/mL | 2000μg/mL |
| G-2 | 1.72±2.04％ | 11.65±2.20％ | 23.26±0.84％ |
| Domperidone derivatives | / | / | 84.74±0.99％ |
| Model set | - | - | - |
The biological activity results show that the intestinal peristalsis effect of the zebra fish in the experimental model group is not improved without any drug treatment, and the intestinal peristalsis effect of the zebra fish is obviously enhanced after the zebra fish is treated by the positive control drug domperidone, and the promotion rate reaches 84.74%. When the concentration of the cistanche tubulosa extract is 200 mug/mL, the effect of promoting the intestinal peristalsis of the zebra fish is not obvious, but when the concentration reaches 667 mug/mL, the intestinal peristalsis of the zebra fish is obviously improved and is more than 10 percent; after the cistanche tubulosa extract with the concentration of 2000 mu g/mL is used for acting, the intestinal tract peristalsis effect of the zebra fish is more remarkable, and the peristalsis promotion rate reaches more than 20%. Therefore, the cistanche tubulosa extract has concentration dependence on the promotion effect of intestinal peristalsis of the zebra fish. In conclusion, the experimental result shows that the cistanche tubulosa extract obtained by the method can obviously promote the intestinal peristalsis and has great potential for developing the prevention or treatment of senile constipation and habitual constipation.
Claims (3)
1. A pharmaceutical composition for preparing a medicament for increasing intestinal motility and preventing or treating senile constipation and habitual constipation, which is characterized in that: the pharmaceutical composition contains a cistanche tubulosa extract and pharmaceutically or edible auxiliary materials;
the extract is prepared by the following method: (1) taking cistanche tubulosa, slicing, and performing reflux extraction for 2-3 times by using water with the mass being 5-10 times of that of the cistanche tubulosa, wherein each time lasts for 2-5 hours; combining the water extracting solutions, carrying out centrifugal filtration, concentrating the filtrate until the density is 1.0-1.2 g/mL (or the solid content is more than 30 wt%) at 40-50 ℃, using the recovered water for next extraction, and cooling the concentrated solution to 30-50 ℃ for later use; (2) stirring the concentrated solution in the step (1), adding alcohol to adjust the volume concentration of the alcohol to be 60-70%, standing overnight, centrifuging, removing supernatant, and drying precipitates to obtain the cistanche tubulosa extract.
2. Use according to claim 1, characterized in that the fresh or dried product of cistanche tubulosa is the above-ground or below-ground part thereof; the alcohol can be one or more of methanol, ethanol, isopropanol or n-butanol, preferably edible or medicinal grade ethanol; the drying mode can be one or more of vacuum drying, normal pressure drying, freeze drying and spray drying.
3. Use according to claim 1, characterized in that: the extract comprises 25-35% of polysaccharide, 9-12% of oligosaccharide, 10-15% of total flavone, 7-12% of total glycosides of cistanche deserticola, 5-8% of polyphenol and 5-8% of protein, and the balance is water and ash based on the mass of the cistanche tubulosa extract.
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