CN101214278A - New raw material for extracting total lignans from Schisandra chinensis, preparation process and application - Google Patents

Abstract

The invention belongs to the technical field of medicine, and discloses a new raw material of a total lignan extract of schisandra chinensis and a preparation process thereof. The process uses the Chinese medicine Schisandra schisandra fruit stem (carp) and Schisandra cotyledon leaves as raw materials, and adopts one or more of processes such as solvent extraction, resin adsorption, normal phase column chromatography, and reverse phase column chromatography after crushing. process to prepare an extract with a total lignan content of 30-95%. The preparation method has the advantages of simple process, stable content and good reproducibility. The extract can replace the total lignan extract of schisandra fruit and be used as medicine or health food, make full use of schisandra resources, and alleviate the situation that the supply of schisandra fruit is in short supply.

Description

New raw material for extracting total lignans from Schisandra chinensis, preparation process and application

technical field

The invention belongs to the technical field of medicine, and relates to a new raw material of Schisandra chinensis total lignan extract and a preparation process thereof. The extract obtained according to the preparation process has a total content of lignans in 30-95%. For the prevention and treatment of hepatitis.

Background technique

The fat-soluble active ingredients of Schisandra chinensis are mainly lignans. Its pharmacological effects involve various aspects such as digestive system, cardiovascular system, central nervous system, reproductive system, anti-oxidation and anti-aging (Chinese herbal medicine, volume 28, phase 2, February 2005).

Schisandra chinensis (Turcz.) Baill., Schisandra chinensis (Turcz.) Baill., Schisandra sphenanthera Rehd.et Wils. and other plants of the genus Schisandra. .

Foreign scholars used high-performance liquid chromatography to analyze the CO ₂ supercritical extract of Schisandra leaves, and found that it contained the same lignans as Schisandra fruit, such as Schizandrin A, Alcohol B, A, B, C and gomisin N et al. (Journal of Chromatography B, 770 (2002) 283289). Schisandra fruit stem (stalk) contains the same lignans as Schisandra fruit, such as schisandrin A, ester A, A, B, etc. (Medical Journal of Yanbian University, March 2005, Volume 28, No. 1).

At present, the market demand for Schisandra chinensis is very large, which is far from meeting the requirements of domestic and foreign markets. Extinct vine harvesting and destruction of wild resources have occurred, and the phenomenon of plundering greens has generally occurred in some areas, which has caused Schisandra chinensis resources to be affected to varying degrees. Destruction, resulting in the reduction of available Schisandra resources, quality decline, and rising prices. When the fruit of Schisandra chinensis is ripe, the fruit and the fruit stem (stalk) are picked together from the vine of Schisandra vine, and the fruit stem (stalk) is removed as impurities, and the resource of Schisandra leaves is very rich. If the leaves and stems ( fruit stalk) as a supplement to the source of Schisandra lignans will be able to make full use of resources and turn waste into treasure.

Contents of the invention

The object of the present invention is to provide two new sources of total lignans of Schisandra chinensis in view of the shortage of above-mentioned Schisandra medicinal materials, the preparation process of two kinds of Schisandra fruit stem (stalk) and leaf total lignan extracts, and the preparation process according to the process The obtained extract and related dosage forms thereof. As a supplement to Schisandra chinensis resources, it can be used as medicine or health food instead of the total lignan extract of Schisandra chinensis fruit.

The present invention is accomplished through the following technical solutions:

(1) Dry Schisandra leaves and fruit stems (fruit stalks), crush them, boil them in water; dry the residue after water extraction, and extract with 50-95% methanol, ethanol, isopropanol or n-butanol under reflux , the extract is concentrated under reduced pressure to a concentrated solution with a density of 0.80-1.20; it is enriched by a macroporous adsorption resin column, eluted with water, 40% to 75% ethanol, and 95% ethanol, and the 95% ethanol eluate is collected and concentrated to obtain The extract of the invention has a total lignan weight content of 30%-95%.

(2) Dry the Schisandra cotyledon and fruit stem (fruit stalk), pulverize, and boil in water; dry the residue after water extraction, and extract with 70-95% methanol, ethanol, isopropanol or n-butanol under reflux The extract is concentrated to dryness under reduced pressure, extracted with an organic solvent at room temperature or heated to reflux, and the extract is eluted with a chromatographic column, and the total lignan weight content of the extract obtained after elution is 30%-95%.

The total lignan extract prepared according to the above process contains compounds: schisandrin A, alcohol B, A, B, C, gomisin G, gomisinL ₁ , gomisinM ₁ , gomisinN and the like. The extract can be mixed with pharmaceutically acceptable carriers to make various clinically available dosage forms.

Beneficial effects of the present invention

The invention aims at replacing the total lignan extract of schisandra fruit as medicine or health food, making full use of schisandra resources, and alleviating the situation that the supply of schisandra fruit is in short supply.

Detailed ways

The following examples further illustrate the invention but do not limit it in any way.

Example 1:

Schisandra schisandra leaves 2Kg, pulverized, refluxed with water to extract twice, each time kept in a slightly boiling state for 1.5 hours, filtered, and the combined filtrate was discarded. After the residue was dried, 14L of 80% ethanol was added, extracted under reflux for 3 times, each time for 2 hours, filtered, and the filtrates were combined and concentrated under reduced pressure to obtain 610 g of a concentrated solution with a density of 0.93. Enriched with a macroporous adsorption resin column, gradient eluted with water, 60% ethanol, and 90% ethanol respectively, collected the 90% ethanol eluate, recovered ethanol under reduced pressure, enriched with a macroporous adsorption resin column again, and used water, 75% ethanol respectively. % ethanol, eluting with 90% ethanol, collecting the 90% ethanol eluate, recovering ethanol under reduced pressure to obtain 15 g of extract. The total lignan content is 60%.

Example 2:

Schisandra schisandra leaves 2Kg, pulverized, refluxed with water to extract twice, each time kept in a slightly boiling state for 1.5 hours, filtered, and the combined filtrate was discarded. After the residue was dried, 14L of 80% ethanol was added, heated and refluxed for 3 times, each time for 2 hours, filtered, the filtrates were combined, and the ethanol was recovered under reduced pressure to obtain 130 g of dry extract. Reflux extraction with petroleum ether for 3 times, combine the extracts, concentrate under reduced pressure to a small volume of 10ml, enrich through silica gel column chromatography, elute with petroleum ether: ethyl acetate (20:1), then use petroleum ether: acetic acid Ethyl ester (2:1) was eluted, and petroleum ether: ethyl acetate (2:1) eluate was collected and concentrated under reduced pressure to obtain 10 g of extract. The total lignan content is 65%.

Example 3:

Schisandra schisandra fruit stem (stalk) 2Kg, pulverized, added water and refluxed for extraction twice, each time kept in a slightly boiling state for 1.5 hours, filtered, and the combined filtrate was discarded. After the residue was dried, 12L of 90% ethanol was added, heated and refluxed for 3 times for 2 hours each time, filtered, and the filtrates were combined and concentrated under reduced pressure to obtain 650 g of a concentrated solution with a density of 0.91. Enriched with a macroporous adsorption resin column, gradient eluted with water, 70% ethanol, and 90% ethanol respectively, collected 90% ethanol eluate, recovered ethanol under reduced pressure to obtain 20 g of extract. The total lignan content is 63%.

Example 4:

Schisandra schisandra fruit stem (stalk) 2Kg, pulverized, added water and refluxed for extraction twice, each time kept in a slightly boiling state for 1.5 hours, filtered, and the combined filtrate was discarded. After the residue was dried, 12L of 90% ethanol was added, heated and refluxed for 3 times for 2 hours each time, filtered, the filtrates were combined and the ethanol was recovered under reduced pressure to obtain 150 g of dry extract. Reflux extraction with petroleum ether for 3 times, combine the extracts, concentrate under reduced pressure to a small volume of 10ml, enrich through silica gel column chromatography, elute with petroleum ether: ethyl acetate (20:1), then use petroleum ether: acetic acid Ethyl ester (2:1) was eluted, and petroleum ether: ethyl acetate (2:1) eluate was collected and concentrated under reduced pressure to obtain 15 g of extract. The total lignan content is 70%.

Example 5:

Schisandra schisandra fruit stem (stalk) 2Kg, pulverized, added water to reflux to extract twice, each time kept in a slightly boiling state for 1.5 hours, filtered, and the combined filtrate was discarded. After the residue was dried, 12L of 90% ethanol was added, heated and refluxed for 3 times for 2 hours each time, filtered, the filtrates were combined and the ethanol was recovered under reduced pressure to obtain 150 g of dry extract. Reflux extraction with petroleum ether for 3 times, combine the extracts, concentrate under reduced pressure to a small volume of 10ml, perform silica gel column chromatography, use petroleum ether: ethyl acetate (100:1 ~ 1:100) gradient elution, combine the same components The eluate was purified by ODS column, high performance liquid phase (HPLC), recrystallization and other methods to obtain schisandra alcohol A, alcohol B, A, B, C, gomisin G, gomisinL ₁ , gomisinM ₁ , gomisinN and so on for 25 compounds. And the use of nuclear magnetic resonance (NMR), mass spectrometry (MS), infrared (IR) and other spectral means to determine their structures.

Embodiment 6:

The Schizandra leaf total lignan extract obtained in Examples 1 and 2 is compared with the compound obtained in Example 5 through thin layer chromatography and high performance liquid phase (HPLC), showing that it contains Schisandrin A, alcohol B, A, B, C, gomisinL ₁ , gomisinM ₁ , gomisinN and other 14 kinds of lignans.

Embodiment 7:

Schisandra schisandra fruit stem (stalk) 2Kg, pulverized, added water and refluxed for extraction twice, each time kept in a slightly boiling state for 1.5 hours, filtered, and the combined filtrate was discarded. After the residue was dried, 12L of 90% ethanol was added, heated and refluxed for 3 times for 2 hours each time, filtered, and the filtrates were combined and concentrated under reduced pressure to obtain 650 g of a concentrated solution with a density of 0.91. Enriched with a macroporous adsorption resin column, gradient eluted with water, 70% ethanol, and 90% ethanol respectively, collected 90% ethanol eluate, recovered ethanol under reduced pressure to obtain 20 g of extract. The total lignan content is 63%. Add 4g of starch, mix evenly, granulate with 80% ethanol, dry, add 0.3g of magnesium stearate, mix evenly, pack into No. 1 capsule, and obtain 70 hard capsules prepared by the present invention. Each capsule contains 0.3g of content.

Example 8: Therapeutic effect of Schisandra chinensis total lignans extract on carbon tetrachloride liver injury mice

Take 50 mice and randomly divide them into 5 groups, which are normal control group, carbon tetrachloride (CCl ₄ ) model group, Schisandra fruit total lignans extract group, Example 1 extract group and Example 3 extract group. Group.

The preparation method of the total lignan extract of Schisandra fruit is as follows: crush the fruit of Schisandra chinensis, add water to reflux and extract twice, keep a slight boiling state for 1.5 hours each time, filter, and discard the combined filtrate. After the residue is dried, add 90% ethanol, heat and reflux to extract 3 times, each time for 2 hours, filter, combine the filtrate and concentrate under reduced pressure, the concentrated solution is enriched with a macroporous adsorption resin column, water, 40% ethanol, 90% Gradient elution with ethanol, collecting 90% ethanol eluate, recovering ethanol under reduced pressure, and recovering ethanol under reduced pressure to obtain an extract with a total lignan content of 70%.

The animal experiment method is as follows: except the normal control group, each group was intraperitoneally injected with 0.1% CCl ₄ paraffin oil solution (10ml/kg) on the first day, and administered by intragastric administration for 7 days continuously at the same time, and blood was taken from the orbit 16 hours after the last administration. Serum alanine aminotransferase (SGPT) was measured.

The experimental results are shown in the table below:

Group (n=10) Dose (mg/kg) SGPT content (U/ml) normal control group 168.1±27.34 CCl ₄ model group 443.2±47.61 Schisandra fruit total lignans extract group 200 271.3±35.53 Example 1 Extract Group 200 281.4±34.79 Example 3 Extract Group 200 287.7±36.88

Conclusion: the SGPT of mice in the CCl4 group was significantly higher than that of the normal control group (P<0.01); Significantly decreased (P<0.01); there was no significant difference among the Schisandra fruit total lignans extract group, the extract group of Example 1, and the extract group of Example 3 (P>0.05). It shows that the total lignan extracts of Schisandra fruit stem (carpone) and leaves and the total lignan extracts of Schisandra fruit have similar therapeutic effects on mice with carbon tetrachloride liver injury, and both have the same effect on mice with carbon tetrachloride liver injury. Elevated SGPT has a significant inhibitory effect.

Claims (10)

1. A new raw material for extracting the total lignans of Schisandra chinensis, characterized in that: the extract obtained by processing the fruit stems and leaves of Schisandra chinensis of Chinese medicine Schisandra chinensis has a total lignans weight content of 30-95%, which can replace the total lignans of Schisandra chinensis fruit Lignan extracts can be used as medicine or health food. 2. The new raw material of total lignans of Schisandra chinensis according to claim 1, characterized in that: its plant source of Schisandra chinensis leaves and fruit stems of Schisandra chinensis is one or both of the following plants: Magnoliaceae Schisandra chinensis Bill. And Schisandrasphenanthera Rehd.et Wils. and other plants of the genus Schisandra. 3. The new raw material for extracting total lignans from Schisandra chinensis according to claim 1 or 2, characterized in that: said lignans are one or more of the following compounds, 1: schisandrin A; 2: Schisandrin B; 3: Schizandrin A; 4: Schizandrin B; 5: Schisandrin C; 6: pseudo-γ-Schisandrin; 7: Neoschisandrin; Phenol B; 11: Schisandra phenol ester; 12: Schisandra ester A; 13: Schisandra ester B; 14: Schisandra ester C; 15: Schisandra ester D; 16: Schisandra ester E; 17: Safflower Schisandra ester; 18: Gomi Sim D; 19: Gomisin E; 20: Gomisin F; 21: Gomisin G; 22: Gomisin H; 23: Gomisin K ₁ ; 24: Gomisin K ₂ ; 25: Ge Misin K ₃ ; 26: Gomisin L ₁ ; 27: Gomisin L ₂ ; 28: Gomisin M ₁ ; 29: Gomisin M ₂ ; 30: Gomisin N; 31: Gomisin 0 32: gomisin R; 33: gomisin H; 34: gomisin 0; 35: gomisin P; 36: gomisin Q; 37: gomisin Misin H; 38: Ciciloylgomisin P; 39: Benzoylgomisin H. 4. a preparation process of Schisandra chinensis total lignans extract as claimed in claim 1, is characterized in that: (1) Schisandra leaves, fruit stems are dried, pulverized, boiled; the residue after water extraction is dried, and A heating and reflux extraction of 50-95% methanol, ethanol, isopropanol or n-butanol, the extract is concentrated under reduced pressure into a concentrated solution with a density of 0.80-1.20 or the extract obtained after the concentrated solution is extracted with an organic solvent. Macroporous adsorption resin column enrichment, eluting with water, 40%-75% ethanol, 95% ethanol, collecting 95% ethanol eluate, and concentrating the extract obtained, the total lignans weight content is 30%-95% %; (2) Dry the Schisandra cotyledon and fruit stem (fruit stalk), pulverize, and boil in water; dry the residue after water extraction, and extract with 70-95% methanol, ethanol, isopropanol or n-butanol under reflux The extract is concentrated to dryness under reduced pressure, extracted with an organic solvent at room temperature or heated to reflux, and the extract is eluted with a chromatographic column, and the total lignan weight content of the extract obtained after elution is 30%-95%. 5. the preparation technology of Schisandra chinensis total lignan extract according to claim 4 is characterized in that: organic solvent is the ether, alcohol, ester, ketone that polarity is less than n-butanol; Solvent elution mode comprises single or mixed Normal phase, reverse phase or gradient elution of solvents. 6. according to the preparation technology of the total lignan extract of Schisandra chinensis according to claim 4, it is characterized in that: described adsorption resin is styrene, phenylpropionic acid, acrylamide or nitric oxide macroporous resin one or more of them. 7. the preparation technology of Schisandra chinensis total lignans extract according to claim 4, is characterized in that: chromatographic column filler is silica gel, aluminum oxide, polyamide, adsorbent resin, glucose gel, ODS, diatomaceous earth one or more. 8. The new raw material for extracting total lignans from Schisandra chinensis according to claim 1, characterized in that: the extract obtained from the raw material can be mixed with pharmaceutically acceptable excipients or carriers to make clinically acceptable Drug and Food Dosage Forms 9. The new raw material for extracting total lignans from Schisandra chinensis according to claim 8, characterized in that: the dosage forms are tablets, capsules, powders, tinctures, oral liquids, dropping pills and injections or beverages, bread, biscuits , candy, wine, tea and cosmetics, etc. 10. The use of new raw materials for extracting total lignans from Schisandra chinensis, characterized in that the extract obtained from the raw materials is used in the preparation of products for preventing or treating hepatitis.