CN101270134B - The preparation method of cucurbitacin - Google Patents

Abstract

The invention provides a method for preparing cucurbitacin. The method is to first extract the pulverized cucurbit tea raw material with an acetone-water solution with a concentration of 40-80%, and the amount of extraction solvent is according to the ratio of material to liquid (g:ml)=1: 6~10, the extraction temperature is 30~80°C, the extraction time is 1.5~9h; after the extract is concentrated and dried, the extract obtained is dissolved in water and filtered, and the filtrate is adsorbed by D ₁₀₁ macroporous resin, and then eluted with deionized water until the effluent has no Then use 10-30% ethanol aqueous solution to elute 6-8 column volumes at a flow rate of 1-2BV/h, collect part of the eluate from the alcohol solution, concentrate and dry to obtain the cucurbitacin product. The preparation method of the invention has good stability, simple and easy operation, low cost, recyclable acetone and ethanol, regenerated macroporous resin, can be used repeatedly, and is suitable for large-scale production.

Description

The preparation method of pericarpium lagenariae theine

Technical field

The present invention relates to extraction of active ingredients preparation method in the natural plant gourds tea, relate in particular to the preparation of polyphenols.

Background technology

The Herba Desmodii Triquetri (Herba Tadehagi Triquetri) prime system extracts the Polyphenols composition that enrichment obtains in Herba Desmodii Triquetri (Herba Tadehagi Triquetri) herb and root.Herba Desmodii Triquetri (Herba Tadehagi Triquetri) Tadehagi triquetrum (Linn.) Ohashi is a pulse family Herba Desmodii Triquetri (Herba Tadehagi Triquetri) platymiscium.This genus has 6 kinds approximately, is distributed in the torrid areas, Asia, Pacific Islands and Australia.China has 2 kinds, i.e. Herba Desmodii Triquetri (Herba Tadehagi Triquetri) and viticula Herba Desmodii Triquetri (Herba Tadehagi Triquetri) T.pseudatriquetrum (DC.) Yang et Huang.Be distributed in ground such as Guangxi, Guangdong, Yunnan, Hainan, Fujian, Jiangxi, Guizhou, Sichuan, Hunan, Taiwan.Record the only Herba Desmodii Triquetri (Herba Tadehagi Triquetri) for medicine, medicinal part is herb (title Herba Desmodii Triquetri (Herba Tadehagi Triquetri)) and root (title Radix Desmodii triquetri), also is that Dai Nationality's medicine and watt medicine are contained.

Herba Desmodii Triquetri (Herba Tadehagi Triquetri): bitter and puckery flavor, cold.Have heat clearing away, dampness removing, function stagnant, parasite killing disappears.Be used for the treatment of flu, pharyngalgia, pneumonopathy hemoptysis, enteritis, dysentery, jaundice, rheumatic arthralgia, ancylostomiasis, vomiting during pregnancy, infantile malnutrition, furuncle.Radix Desmodii triquetri: property is flat, the mildly bitter flavor suffering.Function with heat-clearing and toxic substances removing, dampness removing cough-relieving.Be used for the treatment of the cough due to pathogenic wind-heat, lung abscess, jaundice, carbuncle, scrofula consumptive disease, bone tuberculosis.

Though the Herba Desmodii Triquetri (Herba Tadehagi Triquetri) medicinal history is long, research is very few both at home and abroad.Up to 20 end of the centurys, domesticly just to see report successively, but focus mostly in to farming and animal husbandry aspects such as Mollusca, herding anthelmintics, medical value research is very few.

Contain tannin 7.1～8.6% in the document record calabash Folium Camelliae sinensis, its anti-Mollusca and anthelmintic action cause the interest that we are very big especially.Though because Herba Desmodii Triquetri (Herba Tadehagi Triquetri) research seldom, the research of plant polyphenol is a lot, to medicine, application is very extensive from process hides, food, daily-use chemical industry.Polyphenol is becoming one of focus of current natural product research in research pharmaceutically.Antibacterial, the antiviral of polyphenol, antimutagenic, anticancer, resisting cardiovascular disease, protect the liver kidney tonifying, the anti-ageing curative effect of waiting for a long time is noticeable.Be still the thing that attracts tremendous attention so from natural resources, seek new polyphenol resource.This is rich in the plant resources of polyphenol for further development and use, is necessary extraction, enrichment, the purifying process based on the polyphenol of pericarpium lagenariae theine in Herba Desmodii Triquetri (Herba Tadehagi Triquetri) and the root carried out in depth studying.

In the natural plants in the prior art of extraction of polyphenol substance purifying process, water, alcohol-water, acetone-water extract, the gelatin sedimentation method, the chromatographic isolation analytical method has had more application, but does not also have relevant report at the extraction of polyphenol substance purifying process that in this kind of plant of Herba Desmodii Triquetri (Herba Tadehagi Triquetri) with the pericarpium lagenariae theine is representative.The present invention promptly is intended to finish detailed Herba Desmodii Triquetri (Herba Tadehagi Triquetri) plant polyphenol and extracts purification scheme.

Summary of the invention

The object of the present invention is to provide a kind of complete skill scheme for preparing the high-purity pericarpium lagenariae theine.

The preparation method of pericarpium lagenariae theine of the present invention is to be that 40～80% acetone-water solution extracts the Herba Desmodii Triquetri (Herba Tadehagi Triquetri) raw material after pulverizing with concentration earlier, extract solvent load according to solid-liquid ratio (g: ml)=1: 6～10, extract 30～80 ℃ of temperature, extraction time 1.5～9h; Gained extractum water dissolution, filtration behind the extracting solution concentrate drying, filtrate is through D ₁₀₁After the macroporous resin adsorption, it is colourless to be eluted to effluent with deionized water, uses 10～30% ethanol water then, with 6～8 column volumes of flow velocity eluting of 1～2BV/h, collects alcoholic solution part eluent, and concentrate drying gets the pericarpium lagenariae theine product

The acetone-water solution of wherein said extraction solvent preferred 50～70%, and solid-liquid ratio (g: ml) preferred 1: 8～10, extract preferred 30～50 ℃ of temperature, preferred 4.5～7.5h of extraction time.

In the wherein said purification by macroporous resin step, eluting ethanol water concentration preferred 30%.

The preferred 2BV/h of described ethanol aqueous wash separation of flow speed, preferred 6 column volumes of eluent consumption.

Specifically, the preparation method of pericarpium lagenariae theine of the present invention comprises the steps:

1. be that 50～70% acetone-water solution extracts the Herba Desmodii Triquetri (Herba Tadehagi Triquetri) raw material after pulverizing with concentration, and solid-liquid ratio (g: ml) 1: 8～10, extract 30～50 ℃ of temperature, extraction time 4.5～7.5h;

2. extracting solution concentrate drying, with gained extractum with water dissolution, filtration;

3. filtrate is through D ₁₀₁Macroporous resin adsorption, it is colourless to be eluted to effluent with deionized water then;

4. use 30% ethanol water, with 6 column volumes of flow velocity eluting of 1～2BV/h, collect eluent, concentrate drying gets the pericarpium lagenariae theine product.

In the above-mentioned concrete operating procedure, preferred parameter combinations constitutes the most preferred scheme of the present invention, and is as follows:

1. be that 60% acetone-water solution extracts the Herba Desmodii Triquetri (Herba Tadehagi Triquetri) raw material after pulverizing with concentration, solid-liquid ratio (g: ml) be 1: 8, extract 30 ℃ of temperature, extraction time 6h;

2. extracting solution concentrate drying, with gained extractum with water dissolution, filtration;

3. filtrate is through D ₁₀₁Macroporous resin adsorption, it is colourless to be eluted to effluent with deionized water then;

4. use 30% ethanol water, with 6 column volumes of flow velocity eluting of 1～2BV/h, collect eluent, concentrate drying gets the pericarpium lagenariae theine product.

As the part in the production decision of integrating, the recycling of resin is also extremely important, and the recovery method to the resin after using among the present invention is to adopt with behind 95% the soak with ethanol 24h, cleans repeatedly with ethanol and deionized water.

Preparation method good stability of the present invention, easy to operation, cost is low, and acetone, ethanol are recyclable, and macroporous resin is renewable, all can recycle, and is suitable for large-scale production.

Description of drawings

Accompanying drawing 1 width of cloth of the present invention, i.e. the preparation method flow chart of the pericarpium lagenariae theine of embodiment.

The specific embodiment

The present invention is further illustrated in the mode of specific embodiment below.If no special instructions, measuring method to polyphenol compound among the following embodiment all is standard substance (Nat'l Pharmaceutical ﹠ Biological Products Control Institute with the gallic acid, lot number: 120563-200412, purity 99.1%), adopt colorimetry (Unico7200 visible spectrophotometer, You Nike Shanghai Instr Ltd.) to measure.

The preparation method of purity pericarpium lagenariae theine of the present invention is to put into container after the raw material Herba Desmodii Triquetri (Herba Tadehagi Triquetri) drying and crushing, and (g: ml), extract temperature, time lixiviate, leaching process can stir according to the solvent of setting, solid-liquid ratio; Separation and Extraction liquid also reclaims solvent, and the product lyophilization gets rough pericarpium lagenariae theine.With the Herba Desmodii Triquetri (Herba Tadehagi Triquetri) extracting solution through D ₁₀₁Behind the absorption with macroporous adsorbent resin, it is colourless to be eluted to effluent with deionized water, and carries out eluting with eluent, the eluting material ratio condition set, gathers eluent and reclaims solvent and freezing vacuum drying, gets light yellowish brown graininess pericarpium lagenariae theine.

Following specific embodiment divides other detailed description to extraction and resin purification step.

One, pericarpium lagenariae theine extraction process embodiment

According to above-mentioned concrete operations step, at the extraction pericarpium lagenariae theine down that imposes a condition, extracting solvent is the acetone-aqueous solution of variable concentrations, and yield calculates according to following formula:

Yield=(pericarpium lagenariae theine quality/raw materials quality) * 100%

Actual conditions is set and is extracted and the results are shown in Table 1:

Table 1

Embodiment 21

Present embodiment is an orthogonal experiment of investigating extraction process, and the extraction process step is with aforementioned embodiment 1～20, according to the result of previous embodiment 1～20,4 factors is selected 3 levels suiting respectively, adopts L ₉(3 ⁴) orthogonal experiment calendar investigation optimum process condition.The arrangement of orthogonal experiment factor level sees Table 2, and experimental result sees Table 3 respectively:

Table 2

Table 3

The above result of analysis-by-synthesis, the primary and secondary that influences of each factor is A＞B＞C＞D, promptly extracts temperature＞concentration＞time＞volume.Therefore optimised process combination A1B1C1D2, promptly 8 times of amount 50% acetone water extract 6h for 30 ℃, and the pericarpium lagenariae theine yield is about 6%, is fit to the extraction of pericarpium lagenariae theine.

Two, D ₁₀₁The process conditions of purification by macroporous resin pericarpium lagenariae theine and parameter are investigated embodiment

This part embodiment includes D ₁₀₁Pretreatment and recovery post-processing step before the use of macroporous resin, pretreated method is: get a certain amount of D ₁₀₁Macroporous resin, 95% soak with ethanol 24 hours in its resin column that cleans in advance of packing into, is washed till effluent with 95% ethanol earlier and adds 3 times of no white casse phenomenons of the constant muddiness of water, uses distilled water flushing then instead to there not being the alcohol flavor, can use.Use the method for back recycling to be: behind 5% the soak with ethanol 24h, to clean repeatedly with ethanol and deionized water.

In order to optimize the use of resin, present embodiment is at first investigated the resin property parameter, comprising:

1, determining of static adsorption capacity: in the conical flask of 250ml, add the pretreated D of 5g (dry weight 1.479g) ₁₀₁Macroporous adsorbent resin and 100ml sample solution (concentration is 9.73mg/ml).The room temperature vibration is got 1ml at set intervals and is measured polyphenol content remaining in the solution, draws unit adsorbance and time curve.And according to following formula calculating material balance adsorbance

q＝(C1-C2)V/G

The concentration (mg/ml) of total polyphenols in solution when wherein, C1 and C2 are initial and balance respectively;

V is the volume (ml) of test sample liquid; G is an amount of resin.

Experiment records D ₁₀₁The static adsorption capacity of resin is 272.5mg/ml.

2, the dynamic adsorption amount determines

The investigation of last sample flow velocity: the resin column that to get 4 internal diameters be 2.5cm, the pretreated D of 15g respectively packs into ₁₀₁Macroporous adsorbent resin, adding 50ml concentration respectively is the Herba Desmodii Triquetri (Herba Tadehagi Triquetri) solution of 9.36mg/ml, regulates different flow velocitys, the sample flow velocity is to the influence of absorption with macroporous adsorbent resin pericarpium lagenariae theine in the investigation.Find D according to experimental result ₁₀₁The adsorption capacity of resin is along with the increase of flow velocity slowly descends, when flow velocity is 1～2BV/h, and D ₁₀₁The adsorption capacity of resin changes little, is 103.5mg/g, and when flow velocity was brought up to 4BV/h, its adsorbance reduced to some extent, was 91.8mg/g.Comprehensive post is imitated and the production time cost consideration, and flow velocity selects to be controlled at 1～2BV/h.

Embodiment 22～27

With 50% acetone-water solution serves as to extract solvent, according to solid-liquid ratio 1: 8, extract 30 ℃ of temperature, 6 hours extraction times were extracted pericarpium lagenariae theine from dry Herba Desmodii Triquetri (Herba Tadehagi Triquetri) material powder, extracting solution is used water dissolution, filtration after making extractum, the filtrate upper prop, it is colourless to be eluted to effluent with deionized water after absorption is finished, more respectively according to the eluting that imposes a condition, the enrichment eluent, concentrate drying gets the pericarpium lagenariae theine product, measures polyphenol content (%) in the product, and calculated yield.Actual conditions setting and result are as shown in table 4:

Table 4

Embodiment 28

The present embodiment technological process as shown in Figure 1.

With 50% acetone-water solution serves as to extract solvent, according to solid-liquid ratio 1: 8, extracts 30 ℃ of temperature, and 6 hours extraction times were extracted pericarpium lagenariae theine from dry Herba Desmodii Triquetri (Herba Tadehagi Triquetri) material powder, extraction ratio 6%; Extracting solution is used water dissolution, filtration after making extractum, the filtrate upper prop, and it is colourless to be eluted to effluent with deionized water after absorption is finished, the ethanol of reuse 30% is with 6 column volumes of flow velocity eluting of 2BV/h, enrichment eluent, concentrate drying gets the pericarpium lagenariae theine product, and pericarpium lagenariae theine content is 75%.

Claims (6)

1. A preparation method for cucurbitacin, characterized in that the method is to extract the pulverized cucurbit tea raw material with the acetone-water solution of 40～80% earlier with a concentration, and the extraction solvent consumption is according to the solid-liquid ratio (g:ml)= 1:6~10, extraction temperature 30~80°C, extraction time 1.5~9h; after the extract is concentrated and dried, the extract obtained is dissolved in water and filtered, and the filtrate is absorbed by D ₁₀₁ macroporous resin, and then eluted with deionized water until it flows out The liquid is colorless, and then eluted with 10-30% ethanol aqueous solution at a flow rate of 1-2BV/h for 6-8 column volumes, collecting part of the eluate from the alcohol solution, concentrating and drying to obtain the cucurbitacin product. 2. The preparation method of cucurbitacin according to claim 1, characterized in that the extraction solvent is 50-70% acetone-water solution, the ratio of solid to liquid (g:ml)=1:8-10, extracting The temperature is 30-50°C, and the extraction time is 4.5-7.5 hours. 3. The preparation method of cucurbitacin according to claim 1 or 2, characterized in that in the step of purifying the macroporous resin, the ethanol aqueous solution concentration for eluting is 30%. 4. The method for preparing cucurbitacin according to claim 3, characterized in that the elution flow rate of the ethanol aqueous solution is 2BV/h, and the amount of eluent is 6 column volumes. 5. the preparation method of cucurbitacin according to claim 1 is characterized in that the method is: ①Extract the pulverized gourd tea raw material with acetone-water solution with a concentration of 50-70%, the ratio of extraction solvent to liquid (g:ml)=1:8-10, the extraction temperature is 30-50°C, and the extraction time is 4.5-7.5h ; ②The extract is concentrated and dried, and the obtained extract is dissolved in water and filtered; ③The filtrate was adsorbed by D ₁₀₁ macroporous resin, and then eluted with deionized water until the effluent was colorless; ④Elute 6 column volumes with 30% ethanol aqueous solution at a flow rate of 1-2BV/h, collect the eluate, concentrate and dry to obtain the cucurbitacin product. 6. the preparation method of cucurbitacin according to claim 1 is characterized in that the method is: ① Extract the pulverized gourd tea raw material with a concentration of 60% acetone-water solution, the ratio of extraction solvent to liquid is 1:8, the extraction temperature is 30°C, and the extraction time is 6 hours; ②The extract is concentrated and dried, and the obtained extract is dissolved in water and filtered; ③The filtrate was adsorbed by D ₁₀₁ macroporous resin, and then eluted with deionized water until the effluent was colorless; ④Elute 6 column volumes with 30% ethanol aqueous solution at a flow rate of 1-2BV/h, collect the eluate, concentrate and dry to obtain the cucurbitacin product.

Images