CN107998274B - Ginseng and ophiopogon polysaccharide preparation and its preparing method - Google Patents
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Abstract
The invention discloses a ginseng and ophiopogon polysaccharide preparation and a preparation method thereof, wherein the preparation method comprises the following steps: respectively adding red ginseng and dwarf lilyturf root into ethanol for soaking, then heating and refluxing for extraction, recovering extraction residues, and drying in a dryer to prepare red ginseng medicinal material residues and dwarf lilyturf root medicinal material residues; mixing the red ginseng medicinal residue and the dwarf lilyturf tuber medicinal residue, adding distilled water for extraction, and extracting twice; mixing the extractive solutions, concentrating, and filtering to obtain concentrate; adding absolute ethyl alcohol into the prepared concentrate, standing overnight, centrifuging, collecting precipitate, washing with absolute ethyl alcohol and acetone in sequence, and centrifuging; placing the precipitate in a freeze drying device, vacuum freeze drying, and grinding to obtain Ginseng radix and Maitake polysaccharide powder. The polysaccharide preparation has simple preparation process and low cost, is a reutilization of the extraction residue of the Shenmai injection, and has good immunity enhancing effect.
Description
Technical Field
The invention belongs to the technical field of pharmaceutical preparations, and particularly relates to a ginseng and ophiopogon japonicus polysaccharide preparation and a preparation method thereof.
Background
Ginseng is the dry root of ginseng of Panax genus of Araliaceae, is a perennial herb, is pleasantly cool in the shade, is mainly distributed in Heilongjiang, Jilin, Liaoning provinces and the north deep mountains of Hebei province, and is mostly cultivated artificially at present. The Ginseng radix Rubri is dried root and rhizoma Nelumbinis of steamed Ginseng radix, and is cooked product of Ginseng radix. It is sweet in taste, slightly bitter in taste, warm in nature, and has the effects of invigorating primordial qi, restoring pulse, relieving depletion, benefiting qi, and controlling blood. The red ginseng polysaccharide is one of the important components of red ginseng, and researches show that the red ginseng polysaccharide has certain effects of scavenging free radicals and resisting oxidation in vitro. Modern pharmacological researches find that the Red Ginseng Acidic Polysaccharide (RGAP) has the effects of resisting fatigue, improving climacteric syndrome, inhibiting HIV (human immunodeficiency virus) proliferation, delaying aging, resisting cancer, regulating immunologic function and the like. The reports indicate that the red ginseng polysaccharide can obviously improve the immune function of the mice with low immunity and obviously improve the generation of immune cell factors of the mice with low immunity.
The radix Ophiopogonis is from succulent tuber of Ophiopogon japonicus of Ophiopogon, and has effects of promoting fluid production, quenching thirst, moistening lung, and relieving cough. Radix Ophiopogonis is often combined with other Chinese medicinal herbs to treat dry cough due to lung dryness, thirst due to body fluid consumption, vexation, insomnia, internal heat, diabetes, constipation due to intestinal dryness, etc. Ophiopogonpolysaccharide is one of the important components of ophiopogon root and mainly consists of monosaccharide and oligosaccharide. Modern researches find that the ginseng and ophiopogon polysaccharide has pharmacological effects of resisting oxidation, resisting inflammation, reducing blood sugar and the like. The literature reports that the ophiopogonpolysaccharide can obviously increase the thymus and spleen quality of young mice, can also increase the phagocytic capacity of a reticuloendothelial system of the mice, and can improve the hemolysin content in the serum of the mice and obviously improve the immunity of the mice.
The Shenmai injection is prepared from red ginseng and dwarf lilyturf tuber, and is derived from an ancient formula pulse-activating decoction. Red ginseng tonifies primordial qi, restores pulse, prevents collapse, benefits qi, controls blood, and radix ophiopogonis nourishes yin, promotes fluid production, moistens lung, clears heart, and the two are compatible for tonifying qi and yin and generating yin and yang, and has the efficacy of tonifying qi, preventing collapse, nourishing yin, promoting fluid production, and restoring pulse. The Shenmai injection is widely used for treating shock caused by deficiency of both qi and yin, cardiovascular system diseases such as coronary heart disease, viral myocarditis, chronic pulmonary heart disease, granulocytopenia and the like, can improve the immune function of tumor patients, has certain synergistic effect when being combined with chemotherapeutic drugs, and can reduce toxic and side effects caused by the chemotherapeutic drugs. In 1998, Shenmai injection is listed as a Chinese patent medicine essential for emergency treatment in Chinese medical hospitals.
The Shenmai injection has a wide market in China, but the Shenmai injection is extracted by 90% ethanol, the main components of the Shenmai injection are 7 components of ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, ophiopogonin D', methyl ophiopogonone A and methyl ophiopogonone, polysaccharide components in red ginseng and radix ophiopogonis are not utilized, and the direct discharge of production waste of the Shenmai injection not only causes environmental pollution, but also causes direct waste of cost.
Disclosure of Invention
In view of the above, the present invention provides a ginseng and ophiopogon polysaccharide preparation and a preparation method thereof.
In order to solve the technical problems, the invention discloses a preparation method of a ginseng and ophiopogon polysaccharide preparation, which comprises the following steps:
step 1, removing impurities by using ethanol: respectively soaking Ginseng radix Rubri and radix Ophiopogonis in ethanol, heating and reflux-extracting for 6 times, extracting radix Ophiopogonis for 2 times, and reflux-extracting with 3 times of ethanol each time; recovering extraction residue, and oven drying in a dryer to obtain Ginseng radix Rubri residue and radix Ophiopogonis residue;
step 2, water extraction and concentration: mixing the red ginseng medicinal residue and the dwarf lilyturf tuber medicinal residue, adding distilled water for extraction, and extracting twice; mixing the extractive solutions, concentrating to 1/20 volume, and filtering to obtain concentrate;
and 3, alcohol precipitation: adding absolute ethyl alcohol into the concentrate prepared in the step 2, standing overnight at 4 ℃, centrifuging, collecting precipitate, washing with absolute ethyl alcohol and acetone in sequence, and centrifuging;
step 4, freeze drying to obtain powder: placing the precipitate in a freeze drying device, vacuum freeze drying, and grinding to obtain Ginseng radix and Maitake polysaccharide powder.
Further, the volume fraction of the ethanol in the step 1 is 70-90%; the extraction time is 1.5-2.5 h.
Further, the mass ratio of the red ginseng medicinal material residues to the radix ophiopogonis medicinal material residues in the step 2 is 1: 1; the ratio (g/ml) of the total mass of the red ginseng medicinal material residues and the dwarf lilyturf tuber medicinal material residues to the material-liquid of distilled water is 1:20-1: 40; the water extraction temperature is 60-100 deg.C, and the water extraction time is 0.5-1.5 hr each time.
Further, the volume ratio of the concentrate in the step 3 to the absolute ethyl alcohol is 1:2-1: 6.
The invention also discloses a ginseng and ophiopogon polysaccharide preparation prepared by the preparation method.
Compared with the prior art, the invention can obtain the following technical effects:
1) the polysaccharide preparation obtained by the invention belongs to the field of recycling of waste materials of the ginseng and ophiopogon root injection, a large amount of production waste can be generated in the production of the ginseng and ophiopogon root injection, a large amount of extraction raw materials for extracting the ginseng and ophiopogon root polysaccharide can be collected through resource recycling, the resources are fully utilized, and the cost for producing the ginseng and ophiopogon root polysaccharide preparation is greatly reduced. The polysaccharide preparation has low cost and strong economic benefit.
2) The preparation of the invention only needs basic instruments such as an electric heating jacket (or other heating equipment), a low-speed centrifuge, a drying box, a rotary evaporator and the like, and has simple and reliable process, convenience and easy operation, convenient purchase of the instruments and no field requirement; the related chemical reagents are ethanol and acetone, which are recyclable chemical reagents, the recovery rate can reach more than 95%, the cost is low, no pollutant is generated, and the method is environment-friendly and healthy.
3) The experiment proves that the polysaccharide extraction rate of the polysaccharide extract is 31 percent, and the polysaccharide content of the obtained polysaccharide preparation reaches 62.8 percent.
4) Pharmacodynamic researches show that the product can be used as an immunopotentiator, can obviously enhance thymus and spleen indexes of low-immunity mice, regulate intestinal flora of the low-immunity mice, improve the intestinal immunity of the low-immunity mice, and has a good immunity enhancing effect.
Of course, it is not necessary for any one product in which the invention is practiced to achieve all of the above-described technical effects simultaneously.
Drawings
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the invention and not to limit the invention. In the drawings:
FIG. 1 is a graph of the effect of the present invention's medusa polysaccharide on spleen index in mice with low immunity, where different letters indicate differences between groups, P < 0.01.
Detailed Description
The following embodiments are described in detail with reference to the accompanying drawings, so that how to implement the technical features of the present invention to solve the technical problems and achieve the technical effects can be fully understood and implemented.
The invention discloses a preparation method of a ginseng and ophiopogon polysaccharide preparation, which comprises the following steps:
step 1, removing impurities by using ethanol: soaking red ginseng and radix ophiopogonis in 70-90% ethanol by volume, heating, and extracting under reflux for 6 times and 2 times, wherein 3 times of 70-90% ethanol is used for reflux extraction for 1.5-2.5 h each time; recovering extraction residue, and oven drying in a dryer to obtain Ginseng radix Rubri residue and radix Ophiopogonis residue;
step 2, water extraction and concentration: mixing the red ginseng medicinal material residues and the dwarf lilyturf tuber medicinal material residues according to the ratio of 1: mixing at a ratio of 1(w/w), extracting distilled water at 60-100 deg.C for 0.5-1.5 hr twice at a ratio of 1:20-1:40, mixing the concentrated solutions to 1/20 of the volume of the original extractive solution, and filtering;
and 3, alcohol precipitation: concentrating, adding 2-6 times of anhydrous ethanol (V/V), standing at 4 deg.C overnight, centrifuging, collecting precipitate, washing with anhydrous ethanol and acetone, and centrifuging;
and 4, step 4: freeze drying to obtain powder: placing the precipitate in a freeze drying device, vacuum freeze drying, and grinding to obtain Ginseng radix and Maitake polysaccharide powder.
Example 1
A method for preparing a ginseng and ophiopogon polysaccharide preparation comprises the following steps:
step 1, removing impurities by using ethanol: soaking 100g of radix Ginseng Rubra and 100g of radix Ophiopogonis in 90% (V/V) ethanol respectively, heating, reflux-extracting for 6 times, extracting radix Ophiopogonis for 2 times, and reflux-extracting with 3 times of 90% ethanol for 2 hr each time; recovering extraction residue, and drying in a dryer;
step 2, water extraction and concentration: mixing the residues of the two medicinal materials according to the proportion of 1: mixing at a ratio of 1(w/w), namely mixing distilled water according to a feed-liquid ratio of 1: 30. extracting at 90 deg.C for two times, each for 1 hr, mixing the concentrated solutions to 1/20 volume of the original extractive solution, and filtering;
and 3, alcohol precipitation: concentrating, adding 4 times of anhydrous ethanol (V/V), standing at 4 deg.C overnight, centrifuging, collecting precipitate, washing with anhydrous ethanol and acetone, and centrifuging;
and 4, step 4: freeze drying to obtain powder: placing the precipitate in a freeze drying device, vacuum freeze drying, and grinding to obtain Ginseng radix and Maitake polysaccharide powder.
Example 2
A method for preparing a ginseng and ophiopogon polysaccharide preparation comprises the following steps:
step 1, removing impurities by using ethanol: soaking Ginseng radix Rubri and radix Ophiopogonis respectively in 70% ethanol by volume, heating and reflux-extracting for 6 times, extracting radix Ophiopogonis for 2 times, and reflux-extracting with 3 times of 70% ethanol for 2.5 hr each time; recovering extraction residue, and oven drying in a dryer to obtain Ginseng radix Rubri residue and radix Ophiopogonis residue;
step 2, water extraction and concentration: mixing the red ginseng medicinal material residues and the dwarf lilyturf tuber medicinal material residues according to the ratio of 1: mixing at a ratio of 1(w/w), extracting with distilled water at 100 deg.C for 0.5 hr twice at a ratio of 1:20, mixing the concentrated solutions to 1/20 volume of the original extractive solution, and filtering;
and 3, alcohol precipitation: concentrating, adding 2 times of anhydrous ethanol (V/V), standing at 4 deg.C overnight, centrifuging, collecting precipitate, washing with anhydrous ethanol and acetone, and centrifuging;
and 4, step 4: freeze drying to obtain powder: placing the precipitate in a freeze drying device, vacuum freeze drying, and grinding to obtain Ginseng radix and Maitake polysaccharide powder.
Example 3
A method for preparing a ginseng and ophiopogon polysaccharide preparation comprises the following steps:
step 1, removing impurities by using ethanol: soaking Ginseng radix Rubri and radix Ophiopogonis in 90% ethanol respectively, heating and reflux-extracting for 6 times and 2 times, each time extracting with 3 times of 90% ethanol under reflux for 1.5 hr; recovering extraction residue, and oven drying in a dryer to obtain Ginseng radix Rubri residue and radix Ophiopogonis residue;
step 2, water extraction and concentration: mixing the red ginseng medicinal material residues and the dwarf lilyturf tuber medicinal material residues according to the ratio of 1: mixing at a ratio of 1(w/w), namely extracting distilled water twice at a temperature of 60 ℃ for 1.5h according to a material-liquid ratio of 1:40, combining concentrated solutions to 1/20 of the volume of the original extracting solution, and filtering for later use;
and 3, alcohol precipitation: concentrating, adding 6 times of anhydrous ethanol (V/V), standing at 4 deg.C overnight, centrifuging, collecting precipitate, washing with anhydrous ethanol and acetone, and centrifuging;
and 4, step 4: freeze drying to obtain powder: placing the precipitate in a freeze drying device, vacuum freeze drying, and grinding to obtain Ginseng radix and Maitake polysaccharide powder.
In example 1, the two herb residues were mixed according to the following ratio of 1: mixing at a ratio of 1(w/w), namely mixing distilled water according to a feed-liquid ratio of 1: 30. extracting at 90 deg.C for two times, each for 1 hr, mixing the concentrated solutions to 1/20 volume of the original extractive solution, and filtering; concentrating, adding 4 times of anhydrous ethanol (V/V), standing at 4 deg.C overnight, centrifuging, collecting precipitate, washing with anhydrous ethanol and acetone, and centrifuging; the experiment proves that the polysaccharide content of the obtained polysaccharide preparation reaches 62.8 percent.
In example 2, the red ginseng residue and the dwarf lilyturf tuber residue are mixed according to the ratio of 1: mixing at a ratio of 1(w/w), extracting with distilled water at 100 deg.C for 0.5 hr twice at a ratio of 1:20, mixing the concentrated solutions to 1/20 volume of the original extractive solution, and filtering; concentrating, adding 2 times of anhydrous ethanol (V/V), standing at 4 deg.C overnight, centrifuging, collecting precipitate, washing with anhydrous ethanol and acetone, and centrifuging; the experiment proves that the polysaccharide content of the obtained polysaccharide preparation reaches 60.78 percent.
In example 3, the red ginseng residue and the dwarf lilyturf tuber residue are mixed according to the ratio of 1: mixing at a ratio of 1(w/w), namely extracting distilled water twice at a temperature of 60 ℃ for 1.5h according to a material-liquid ratio of 1:40, combining concentrated solutions to 1/20 of the volume of the original extracting solution, and filtering for later use; concentrating, adding 6 times of anhydrous ethanol (V/V), standing at 4 deg.C overnight, centrifuging, collecting precipitate, washing with anhydrous ethanol and acetone, and centrifuging; tests prove that the polysaccharide content of the obtained polysaccharide preparation reaches 61.36 percent.
It can be seen that the red ginseng medicinal residue and the dwarf lilyturf tuber medicinal residue are mixed according to the proportion of 1: 1(w/w), mixing distilled water according to a feed-liquid ratio of 1: 30. extracting at 90 deg.C for two times, each for 1 hr, mixing the concentrated solutions to 1/20 volume of the original extractive solution, and filtering; concentrating, adding 4 times of anhydrous ethanol (V/V), standing at 4 deg.C overnight, centrifuging to collect precipitate, washing with anhydrous ethanol and acetone, and centrifuging to obtain polysaccharide preparation with highest content. Therefore, the ginseng and wheat polysaccharide powder with the highest polysaccharide content can be obtained according to the extraction and alcohol precipitation method in the first embodiment.
Example 4
The invention uses the ginseng polysaccharide in mice with low immunity, and carries out the following effect experiments on the influence of immune organ index, intestinal immune factors and intestinal flora of the mice:
selecting 40 mice of 18-22g, randomly grouping into a blank group, a model group, a high-dose group and a low-dose group, constructing a mouse immune hypofunction model by using cyclophosphamide, injecting 60mg/kg of cyclophosphamide into the mice through intraperitoneal injection for three days, wherein the blank group is injected with physiological saline.
And (3) after the molding is finished, performing intragastric administration on the mice, performing intragastric administration on the blank group and the model group by using normal saline, performing intragastric administration on the mice in the high-dose group by using 100mg/kg panaxan powder samples, performing intragastric administration on the mice in the low-dose group by using 50mg/kg panaxan powder samples, and performing intragastric administration for 7 days.
Weighing the mice the next day after the completion of the gavage, killing the mice after neck fracture, dissecting the mice to take spleen and thymus organs, calculating immune organ indexes after weighing and recording, taking out intestinal tract tissues of the mice, separating the cecum contents and ileum tissues of the mice, and storing for later use. Wherein, the caecum content is used for detecting the intestinal flora, and the ileum tissue is used for detecting the intestinal immunity protein SIgA.
The detection result shows that the thymus index and spleen index of the mice are obviously reduced after the mice are modeled, and the thymus index and spleen index of the mice in a high-dose group and the mice in a low-dose group are obviously increased after the mice are gavaged. In spleen index results, the value of the dose group is obviously higher than that of the model group, the three groups have obvious difference, P is less than 0.01, and the high dose group is obviously higher than the low dose, so that the high dose of the ginseng polysaccharide powder has obvious effect of enhancing immune organs. In thymus index results, the values of the dose groups are obviously higher than those of the model groups, the high dose groups and the model groups have obvious difference, P is less than 0.05, and the effect is more obvious compared with that of the low dose groups. See table 1 and table 2 for details.
Through the determination of the SIgA content of ileum tissues, detection results show that the Shenmai polysaccharide powder obviously improves the content of intestinal immune secretory protein SIgA, the SIgA content of a dose group is obviously higher than that of a model group, and the dosage group has significant difference with the model group, wherein P is less than 0.05. The ginseng and ophiopogon polysaccharide powder can obviously improve the content of intestinal immune protein SIgA of mice with low immunity to a certain extent. See table 3 for details.
The intestinal flora result detection shows that after the modeling is finished, the quantity of bifidobacteria and lactobacilli in intestinal tracts of mice is obviously reduced, and is obviously different from that of a blank group, and P is less than 0.05. After the ginseng and ophiopogon polysaccharide powder is used for gastric lavage, the quantity of bifidobacteria in a dose group is obviously increased, and the quantity of bifidobacteria in a high dose group is higher. In lactobacillus detection, the number of lactobacillus in the dose group is higher than that in the model group and higher than that in the blank group, and the dose group has a significant difference with the model group and the blank group, wherein P is less than 0.05. The results show that the panaxan powder has obvious enhancement effect on immune organs of mice with low immunity, obviously influences intestinal immunity and improves the intestinal immunity of the mice with low immunity. See table 4 for details.
TABLE 1 Effect of Panama polysaccharide on spleen index in immunocompromised mice
Note: in the table, different letters indicate differences between groups, P < 0.05.
TABLE 3 influence of Ginseng polysaccharide on the content of SIGA in intestinal tract of immunocompromised mice
Note: in the table, different letters indicate differences between groups, P < 0.05.
TABLE 4 influence of Ginseng polysaccharide on intestinal flora of immunocompromised mice
Note: in the table, different letters indicate differences between groups, P < 0.05.
While the foregoing description shows and describes several preferred embodiments of the invention, it is to be understood, as noted above, that the invention is not limited to the forms disclosed herein, but is not to be construed as excluding other embodiments and is capable of use in various other combinations, modifications, and environments and is capable of changes within the scope of the inventive concept as expressed herein, commensurate with the above teachings, or the skill or knowledge of the relevant art. And that modifications and variations may be effected by those skilled in the art without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (2)
1. The preparation method of the ginseng and ophiopogon japonicus polysaccharide preparation is characterized by comprising the following steps:
step 1, removing impurities by using ethanol: respectively soaking Ginseng radix Rubri and radix Ophiopogonis in ethanol, heating and reflux-extracting for 6 times, extracting radix Ophiopogonis for 2 times, and reflux-extracting with 3 times of ethanol each time; recovering extraction residue, and oven drying in a dryer to obtain Ginseng radix Rubri residue and radix Ophiopogonis residue;
step 2, water extraction and concentration: mixing the red ginseng medicinal residue and the dwarf lilyturf tuber medicinal residue, adding distilled water for extraction, and extracting twice; mixing the extractive solutions, concentrating to 1/20 volume, and filtering to obtain concentrate;
and 3, alcohol precipitation: adding absolute ethyl alcohol into the concentrate prepared in the step 2, standing overnight at 4 ℃, centrifuging, collecting precipitate, washing with absolute ethyl alcohol and acetone in sequence, and centrifuging;
step 4, freeze drying to obtain powder: placing the precipitate in a freeze drying device, vacuum freeze drying, and grinding to obtain radix Ginseng and Maifanitum polysaccharide powder;
the volume fraction of the ethanol in the step 1 is 70-90%; the extraction time is 1.5h-2.5 h;
the mass ratio of the red ginseng medicinal material residues to the dwarf lilyturf tuber medicinal material residues in the step 2 is 1: 1; the ratio (g/ml) of the total mass of the red ginseng medicinal material residues and the dwarf lilyturf tuber medicinal material residues to the material-liquid of distilled water is 1:20-1: 40; the water extraction temperature is 60-100 ℃, and the water extraction time is 0.5-1.5h each time;
the volume ratio of the concentrate in the step 3 to the absolute ethyl alcohol is 1:2-1: 6.
2. A ginseng polysaccharide preparation prepared by the preparation method of claim 1.
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| CN108969649A (en) * | 2018-09-17 | 2018-12-11 | 四川农业大学 | One seed ginseng wheat polyoses oral liquid and preparation method thereof |
| CN110423286A (en) * | 2019-08-30 | 2019-11-08 | 鲁南制药集团股份有限公司 | A kind of extracting method and application thereof of Chinese medicine dreg polysaccharide |
| CN112830977B (en) * | 2021-02-26 | 2022-05-17 | 西南科技大学 | Method for extracting and purifying polysaccharide from fibrous root residues of radix ophiopogonis |
| CN114668075A (en) * | 2022-04-20 | 2022-06-28 | 四川农业大学 | Ginseng and wheat fibrous root polysaccharide granules and preparation method and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1596954A (en) * | 2004-09-24 | 2005-03-23 | 贵阳云岩西创药物科技开发有限公司 | Red ginseng and ophiopogon root preparation and its making method |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1596954A (en) * | 2004-09-24 | 2005-03-23 | 贵阳云岩西创药物科技开发有限公司 | Red ginseng and ophiopogon root preparation and its making method |
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| Title |
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| 参麦有效部位抗肿瘤作用的实验研究;方三华;《中国优秀博硕士学位论文全文数据库（硕士） 医药卫生科技辑》;20040915(第3期);5-6、19-20 * |
| 生脉散多糖的组成及其初级结构分析;胡闻莉等;《中国药科大学学报》;20021231;第33卷(第1期);38-41 * |
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