CN105878518A - Separation and purification method of litchi polyphenol - Google Patents

Abstract

The invention provides a method for separation and purification of litchi polyphenols. The extraction process is as follows: transfer all the crushed litchis to a beaker, add distilled water with a solid-liquid ratio of 1:3-8g/ml, and immerse the litchis completely at 100°C. After boiling for 20‑40min, lower the temperature to 60‑80°C and soak for 2‑6h, filter and collect the filtrate; then wash the filter residue with warm water, repeat at least 2 times, and collect the filtrate obtained each time; then add acetone to the filter residue solution, and soaked at 50-70°C for 6-10h, filtered again and collected the filtrate, and finally all the filtrates were combined and dried to obtain the crude extract of litchi polyphenols; subsequently, purified with ethyl acetate in ethanol solution to obtain the purified intermediate The product is adsorbed and separated by ADS‑7D macroporous resin. The separation and purification method of litchi polyphenols is simple to operate, fully uses discarded litchi skins and litchi cores to extract litchi polyphenols, the extraction amount is high, and the purity is high. In addition, the extraction process does not require special instruments, and the cost is low. It is safe to use and improves The economic value of litchi.

Description

A kind of separation and purification method of litchi polyphenol

technical field

The invention relates to the technical field of extraction technology, in particular to a method for separating and purifying litchi polyphenols.

Background technique

Litchi is the fruit of Sapindaceae plant Litchi, which originates in southern my country and is most cultivated in Guangdong, Guangxi, Fujian, Sichuan, Taiwan, Yunnan and other places. Litchi has high nutritional value and is rich in sugar and VC. The medicinal value of lychee is also high. It is a statutory dual-purpose plant for medicine and food announced by the Ministry of Health. It is warm in nature, sweet and sour. , Nourish the liver, regulate qi, and benefit blood; while lychee core has the effect of warming the middle, regulating qi and relieving pain.

Litchi contains a large amount of polyphenols, mainly polyphenols. Polyphenols refer to a group of chemical elements in plants, named after having multiple phenolic groups. Polyphenols have a strong antioxidant effect. Oriental tea and western red wine contain polyphenols. Polyphenols can prevent the aging of the human body and brain, and have anticancer activity and high economic value. The extraction and purification methods of polyphenols have made great progress, but the current common purification process cannot effectively extract polyphenols from litchi.

Therefore, prior art still needs further improvement.

Contents of the invention

In view of the deficiencies in the prior art, the present invention provides a method for separating and purifying litchi polyphenols, aiming at solving the problem that there is no corresponding technology that can effectively extract polyphenols from litchi.

A method for separating and purifying litchi polyphenols, wherein the method is:

Extraction process: Transfer all the crushed lychees to a beaker, add distilled water with a solid-liquid ratio of 1:3-8g/ml, immerse all the lychees, boil at 100°C for 20-40min, then lower the temperature to 60-80°C Soak for 2-6 hours, filter and collect the filtrate; then wash the filter residue with warm water, repeat at least 2 times, and collect the filtrate obtained each time; then add acetone solution to the filter residue respectively, and keep soaking at 50-70°C for 6-10h, and again Filtrating and collecting the filtrate, finally combining all the filtrates and drying them to obtain the crude extract of litchi polyphenols;

Purification process: After dissolving the crude polyphenol extract of litchi with ethanol solution, add ethyl acetate equal to the ethanol solution, let it stand at room temperature, and separate the ethyl acetate layer and the ethanol layer after liquid layering; then use ethyl acetate The ethanol layer was extracted with ester, and after separation, the ethanol layer was evaporated to dryness to obtain a dry purified intermediate product;

After the obtained purified intermediate product is dissolved in water and filtered, the purified intermediate product is subjected to adsorption and separation through ADS-7D macroporous resin.

The method for separating and purifying litchi polyphenols, wherein an acetone solution with a solid-to-liquid ratio of 1:3-8 g/ml is added during the extraction process.

The method for separating and purifying litchi polyphenols, wherein the litchi fragments are fragments of litchi core or litchi shell.

The method for separating and purifying litchi polyphenols, wherein, during the extraction process, after boiling at 100° C. for 30 minutes, the temperature is lowered to 70° C. and soaked for 4 hours.

The separation and purification method of litchi polyphenols, wherein the concentration of the acetone solution used is 50%.

The method for separating and purifying litchi polyphenols, wherein, during the extraction process, soaking in acetone solution at 60°C for 8 hours.

The separation and purification method of litchi polyphenols, wherein the concentration of the ethanol solution used in the purification process is 20%.

The separation and purification method of litchi polyphenols, wherein, in the purification process, an ethanol solution with a solid-liquid ratio of 1:8-12g/ml is added.

The method for separating and purifying litchi polyphenols, wherein the method for purifying with a macroporous adsorption resin is specifically: according to the ratio of purified intermediate product: wet weight of resin about 1:2-6, place the processed Add the purified intermediate product into the beaker of resin ADS-7, shake it every 5 minutes for 10 seconds, last for 2 hours; then let it stand for 24 hours to make it reach saturated adsorption, pour out the eluent, and then wash the macroporous resin with ethanol and distilled water respectively Remove about 3-4 times, filter to obtain water extract, and dry.

The method for separating and purifying litchi polyphenols, wherein the wet weight ratio of the purified intermediate product to the macroporous adsorption resin is 1:4.

Beneficial effects: the present invention provides a method for separation and purification of litchi polyphenols. The separation and purification method does not require special instruments, is widely used, and has low cost. The experiment is economical, easy to obtain, and safe to use. The extraction of litchi polyphenols The content is high and the purity is high, the oxidation degree of melanin is low, and the extraction period is short, and it is suitable for large-scale extraction of litchi polyphenols, which improves the economic value of litchi.

detailed description

The present invention provides a separation and purification method of litchi polyphenols. In order to make the purpose, technical scheme and effect of the present invention clearer and clearer, the present invention will be further described in detail below. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

The invention provides a method for separating and purifying litchi polyphenols, the method for separating and purifying is:

A method for separating and purifying litchi polyphenols, wherein the method is:

Extraction process: Transfer all the crushed lychees to a beaker, add distilled water with a solid-liquid ratio of 1:3-8g/ml, immerse all the lychees, boil at 100°C for 20-40min, then lower the temperature to 60-80°C Soak for 2-6 hours, filter and collect the filtrate; then wash the filter residue with warm water, repeat at least 2 times, and collect the filtrate obtained each time; then add acetone solution to the filter residue respectively, and keep soaking at 50-70°C for 6-10h, and again Filtrate and collect the filtrate, finally combine all the filtrates and dry to obtain the crude extract of litchi polyphenols.

The litchi fragments are fragments of litchi cores or litchi shells. Because the polyphenol content in the litchi shell core is high, it is a good raw material for extracting polyphenols, and at the same time, waste can be utilized to improve the utilization rate and economic value of the litchi.

During the extraction process, distilled water is used to extract polyphenols from litchi, which can effectively avoid the introduction of microorganisms and impurities into the extract, thereby reducing the problem of microbial contamination.

Boiling the extract at 100°C for 20-40 minutes can effectively destroy the cell wall of the litchi cells, allowing the polyphenols in the cells to diffuse into the water faster, and the time is controlled within 20-40 minutes, which can also effectively prevent excessive impurities from dissolving into the water , and avoid the denaturation of polyphenols. Preferably, when boiling at 100° C. for 30 minutes, the efficiency of polyphenol dissolution is the highest.

After boiling and leaching, lower the temperature to 70°C and soak for 4 hours. At this time, lowering the temperature and performing leaching for 4 hours can better extract polyphenols from litchi without affecting the activity of polyphenols.

Subsequently, the filter residue was washed with warm water at least twice to extract more polyphenols from the filter residue.

In the further extraction process, the purpose of adding acetone solution with a solid-liquid ratio of 1:3-8g/ml is to reclaim the polyphenols easily soluble in acetone in the litchi filter residue and increase the extraction amount of polyphenols. When the concentration of the acetone solution used was 50%, the amount of polyphenols extracted was the largest.

Preferably, soaking in 50% acetone solution at 60°C for 8 hours will result in the most polyphenols in litchi.

Purification process: after dissolving the crude polyphenol extract of litchi with ethanol solution, add ethyl acetate equal to the ethanol solution, let it stand at room temperature, separate the ethyl acetate layer and the ethanol layer after liquid layering; then use ethyl acetate The ethanol layer was extracted with ester, and after separation, the ethanol layer was evaporated to dryness to obtain a dry and purified intermediate product.

Wherein, the concentration of the ethanol solution used in the purification process is 20%. Polyphenols have the highest solubility in 20% ethanol solution, while polyphenols are insoluble in ethyl acetate. Therefore, using these two solutions to separate crude polyphenols from other impurities has a good purification effect.

The method for separating and purifying litchi polyphenols, wherein an ethanol solution with a solid-to-liquid ratio of 1:8-12 g/ml is added during the purification process. The preferred solid-liquid ratio is 1:10g/ml. Under this condition, the extraction efficiency of litchi polyphenols is the highest.

After the obtained purified intermediate product is dissolved in water and filtered, the purified intermediate product is subjected to adsorption and separation through ADS-7D macroporous resin. ADS-7D resin has the best selective adsorption effect on lychee flavonoids, which can greatly improve the purification efficiency.

Wherein, the method for purifying with macroporous adsorption resin is specifically: according to the ratio of purified intermediate product: wet weight of resin about 1:2-6, add the purified intermediate product to the beaker where the treated resin ADS-7D is placed , shake every 5min for 10s for 2h; then let it stand for 24h to achieve saturated adsorption, pour out the eluent, then elute the macroporous resin with ethanol and distilled water for about 3-4 times, filter to obtain the water extract objects, to dry.

In a preferred embodiment, the purification intermediate product: when the wet weight of the resin is about 1:2-6, the macroporous resin has a good adsorption effect on litchi polyphenols, and can adsorb polyphenols in the solution to the greatest extent. Preferably, the wet weight ratio of the purified intermediate product to the macroporous adsorption resin is 1:4.

When 20% ethanol solution and distilled water are used to elute the macroporous resin during the purification process, the system effect of 20% ethanol solution and distilled water on the polyphenols adsorbed on the resin is the best, and the polyphenols can be eluted to the greatest extent.

In order to describe the preparation method of the present invention in more detail, several examples are listed below for further illustration.

Example 1

The separation and purification method of litchi polyphenols provided by the invention is as follows:

Extraction process: Transfer all the crushed lychees to a beaker, add 1:8g/ml distilled water to fully immerse the lychee skin under the liquid surface; boil and keep soaking at 100°C for 0.5h, then lower the temperature to After soaking at 70°C for 4 hours, filter and collect the filtrate; wash the filter residue with warm water, repeat 3 times, and collect the filtrate obtained each time; Soak for 8 hours at ℃, filter again and collect the filtrate, and finally combine all the filtrates and dry to obtain the crude extract of litchi polyphenols;

Purification process: Dissolve the crude polyphenol extract of lychee with a 20% ethanol solution with a material-to-liquid ratio of 1:10g/ml, add ethyl acetate equal to the ethanol solution and let it stand for about 10 minutes, place it at room temperature, and separate the liquid , separated by a separatory funnel to obtain an ethyl acetate layer and an ethanol layer; then extract the ethanol layer with 10 ml of ethyl acetate, evaporate the ethanol layer to dryness after separation, and obtain a dry purified intermediate product.

According to the ratio of purified intermediate product: resin wet weight about 1:4, add the purified intermediate product into the beaker with the treated resin ADS-7, shake it every 5min for 10s for 2h; then let it stand for 24h to make it When saturated adsorption is reached, the eluent is poured out, and then the macroporous resin is eluted with 20% ethanol and distilled water for about 3-4 times, and the water extract is obtained by filtration and dried.

Experimental results: The crude polyphenol contents are: litchi skin 213.96ug/g, litchi core 209.33ug/g.

Example 2

The separation and purification method of litchi polyphenols provided by the invention is as follows:

Extraction process: Transfer all the crushed lychees to a beaker, add 1:5g/ml distilled water to fully immerse the lychee skin under the liquid surface; boil and keep soaking at 100°C for 20 minutes, then lower the temperature to 60°C After soaking at ℃ for 2 hours, filter and collect the filtrate; wash the filter residue with warm water, repeat twice, and collect the filtrate obtained each time; then add 1:8g/ml 50% acetone solution to the filter residue respectively, and keep at 50°C Soak for 6h, filter again and collect the filtrate, finally combine all the filtrates and dry to obtain the crude extract of litchi polyphenols;

Purification process: Dissolve the crude polyphenol extract of lychee with a 20% ethanol solution with a material-to-liquid ratio of 1:8g/ml, add ethyl acetate equal to the ethanol solution and let it stand for about 10 minutes, place it at room temperature, and separate the liquid , separated by a separatory funnel to obtain an ethyl acetate layer and an ethanol layer; then extract the ethanol layer with 10 ml of ethyl acetate, evaporate the ethanol layer to dryness after separation, and obtain a dry purified intermediate product.

According to the ratio of purified intermediate product: resin wet weight about 1:2, add the purified intermediate product into the beaker with the treated resin ADS-7, shake it every 5min for 10s for 2h; then let it stand for 24h to make it When saturated adsorption is reached, the eluent is poured out, and then the macroporous resin is eluted with 20% ethanol and distilled water for about 3-4 times, and the water extract is obtained by filtration and dried.

Experimental results: The crude polyphenol content is: litchi skin 210.56ug/g, litchi core 205.23ug/g.

Example 3

The separation and purification method of litchi polyphenols provided by the invention is as follows:

Extraction process: Transfer all the crushed lychees to a beaker, add 1:3g/ml distilled water to fully immerse the shredded lychees under the liquid surface; boil and soak at 100°C for 40 minutes, then lower the temperature to 80°C After soaking at ℃ for 6 hours, filter and collect the filtrate; wash the filter residue with warm water, repeat 3 times, and collect the filtrate obtained each time; then add 1:3g/ml 50% acetone solution to the filter residue respectively, and keep at 70°C Soak for 10 hours, filter again and collect the filtrate, finally combine all the filtrates and dry to obtain the crude extract of litchi polyphenols;

Purification process: After dissolving the crude polyphenol extract of litchi with 20% ethanol solution with a material-to-liquid ratio of 1:12g/ml, add ethyl acetate equal to the ethanol solution and let it stand for about 10 minutes, place it at room temperature, and separate the liquid , separated by a separatory funnel to obtain an ethyl acetate layer and an ethanol layer; then extract the ethanol layer with 10 ml of ethyl acetate, evaporate the ethanol layer to dryness after separation, and obtain a dry purified intermediate product.

According to the ratio of purified intermediate product: resin wet weight about 1:6, add the purified intermediate product into the beaker with the treated resin ADS-7, shake it every 5min for 10s for 2h; then let it stand for 24h to make it When saturated adsorption is reached, the eluent is poured out, and then the macroporous resin is eluted with 20% ethanol and distilled water for about 3-4 times, and the water extract is obtained by filtration and dried.

Experimental results: The crude polyphenol content is: litchi skin 208.15.ug/g, litchi core 201.24ug/g.

The separation and purification method of litchi polyphenols is simple to operate, fully uses discarded litchi skins and litchi cores to extract litchi polyphenols, the extraction amount is high, and the purity is high. In addition, the extraction process does not require special instruments, and the cost is low. It is safe to use and improves The economic value of litchi.

It should be understood that the application of the present invention is not limited to the above examples, and those skilled in the art can make improvements or transformations according to the above descriptions, and all these improvements and transformations should belong to the protection scope of the appended claims of the present invention.

Claims (10)

1. the isolation and purification method of a lichee polyphenol, it is characterised in that described method is:

Extraction process: the lichee after pulverizing is transferred completely in beaker, adding solid-liquid ratio is the distilled water of 1:3-8g/ml, entirely Portion's submergence lichee, after 100 DEG C are boiled 20-40min, cools the temperature to 60-80 DEG C and soaks 2-6h, filter and collect filtrate；Use again Warm water washing filter residue, is at least repeated 2 times, collects the filtrate every time obtained；Acetone soln is each added the most again in filter residue, and Keep 50-70 DEG C to soak 6-10h, again filter and collect filtrate, finally all filtrates being merged, be dried, obtain lichee Polyphenol crude extract；

Purge process: after being dissolved by lichee polyphenol crude extract with ethanol solution, after adding the ethyl acetate with ethanol solution equivalent Stand, place room temperature, separating ethyl acetate layer and alcohol layer after liquid layered；It is extracted with ethyl acetate alcohol layer again, after separation Alcohol layer is volatilized, obtains the purification of intermediates being dried；

After the purification of intermediates obtained being dissolved in water and filtering, by purification of intermediates through the macroreticular resin of ADS-7D specification Carry out adsorbing separation.

The isolation and purification method of lichee polyphenol the most according to claim 1, it is characterised in that add feed liquid during extraction Than the acetone soln for 1:3-8g/ml.

The isolation and purification method of lichee polyphenol the most according to claim 1, it is characterised in that described lichee fragment is lichee Core or the fragment of lychee exocarp.

The isolation and purification method of lichee polyphenol the most according to claim 1, it is characterised in that during extraction, 100 DEG C are boiled After boiling 30min, cool the temperature to 70 DEG C and soak 4h.

The isolation and purification method of lichee polyphenol the most according to claim 1, it is characterised in that the concentration of acetone soln used It is 50%.

The isolation and purification method of lichee polyphenol the most according to claim 1, it is characterised in that during the described extraction of institute, In acetone soln, keep 60 DEG C soak 8h.

The isolation and purification method of lichee polyphenol the most according to claim 1, it is characterised in that institute in the described purge process of institute The concentration of ethanol solution be 20%.

The isolation and purification method of lichee polyphenol the most according to claim 1, it is characterised in that add in the described purge process of institute Enter the ethanol solution that solid-liquid ratio is 1:8-12g/ml.

The isolation and purification method of lichee polyphenol the most according to claim 1, it is characterised in that described macroporous absorbent resin The method that is purified is particularly as follows: according to purification of intermediates: the ratio of resin weight in wet base about 1:2-6, handles well to being placed with The beaker of Resin A DS-7 adds purification of intermediates, shakes 10s every 5min, continue 2h;Then 24h is stood so that it is reach Saturated absorption, pours out eluent, then is eluted about 3-4 time with ethanol and distilled water respectively by macroreticular resin, be filtrated to get water and carry Thing, is dried.

The isolation and purification method of lichee polyphenol the most according to claim 9, it is characterised in that described purification of intermediates It is 1:4 with the weight in wet base ratio of macroporous absorbent resin.