CN102526315B - Preparation method of extracts of effective fractions of lychee seeds - Google Patents

Abstract

The invention belongs to the technical field of extraction and purification of traditional Chinese medicines, and more specifically relates to a preparation method of an extract of effective parts of litchi kernels from a traditional Chinese medicine, comprising the following steps: (1) crushing litchi kernels medicinal material, adding ethanol for ultrasonic extraction; (2) recovering the extract Dilute the ethanol with water, and filter after standing; (3) the filtrate is adsorbed by AB-8 macroporous resin, eluted with water first, followed by ethanol; (4) collect the ethanol eluate, recover ethanol, continue to concentrate until thick paste; (5) dry the extract to obtain. The present invention utilizes the full extraction effect of ethanol on the active ingredients of litchi core and the selective effect of macroporous resin with high selectivity to simultaneously obtain the extract of total flavonoids, total saponins and effective parts of tannin in litchi core, and the sum of the contents of the three Reaching more than 70%, the process is simple and stable, and the quality is controllable, and the obtained litchi core effective part group extract meets the requirements of the theory of traditional Chinese medicine and the modernization of traditional Chinese medicine.

Description

A kind of preparation method of effective part group extract of litchi core

technical field

The invention belongs to the technical field of extraction and purification of traditional Chinese medicines, and in particular relates to a preparation method of extracts of effective fractions of litchi kernels of traditional Chinese medicines.

Background technique

Lychee Seed, Litchi Seed, is the dried and mature seed of Sapindaceae plant Litchi (Litchi Chinensis Sonn), also known as Li Ren, Li He, Da Li He, recorded in "Chinese Pharmacopoeia" 2010 edition one, Warm in nature, sweet in taste, slightly bitter, has the effects of promoting qi and dispelling stagnation, dispelling cold and relieving pain. "Compendium of Materia Medica" records that lychee core "quenches thirst, benefits people's color, and eats it to relieve polydipsia" (polydipsia, modern medicine is called diabetes). Studies have shown that lychee core has the functions of lowering blood sugar, regulating blood lipids, improving anti-oxidation, and inhibiting the surface antigen of hepatitis B virus. The ingredients contained in lychee kernels have been well studied. The main ingredients are saponins, flavonoids and anthocyanins, phytosterols, tannins and polyphenols, fatty acids, amino acids, proteins and polysaccharides. Experimental research and clinical treatment have shown that lychee core extract can significantly reduce blood sugar and blood lipids, and effectively treat diabetes and metabolic syndrome. Making full use of the abundant local litchi kernel resources to develop safe, effective and quality-controllable drugs for the treatment of diabetes for clinical application has important social significance in changing the trend of high prevalence of diabetes caused by lifestyle changes and market value.

The existing litchi kernel extraction and purification processes include water extraction and alcohol precipitation method, ethanol extraction-macroporous resin purification method, etc., which mostly target a single active ingredient or part, and remove the rest of the ingredients as impurities, and the obtained extract does not meet the requirements. Theory of Traditional Chinese Medicine. "Effective part" refers to a class of chemical components extracted from single Chinese herbal medicines or decoction pieces that have been proved effective by animal and clinical tests. Compared with the original medicinal materials, it is enriched with effective substances, not only takes a small amount, but also improves the curative effect. One of the characteristics of traditional Chinese medicine is compound medicine. The characteristics of traditional Chinese medicine are multi-target, multi-site and multi-level comprehensive effects, and the corresponding material basis of action should also be multi-component rather than single component. Litchi, litchi kernels and their effective parts have pharmacological and pharmacodynamic effects such as lowering blood sugar, regulating blood lipids, anti-oxidation, inhibiting viruses, anti-tumor and anti-liver damage. Among them, flavonoids can inhibit virus and anti-tumor, total saponins can inhibit virus activity, lower blood sugar, regulate blood lipids and enhance insulin sensitivity, flavonoids, total saponins and polysaccharides have antioxidant effects, polysaccharides can improve immune function . Combining effective parts derived from different raw materials into a compound, that is, "effective part group" for clinical use, not only reflects the characteristics of traditional Chinese medicine, but also improves the technical content, which is also the need to realize the modernization and internationalization of traditional Chinese medicine.

"Technology Research on the Purification of Total Saponins of Litchi Seed by Macroporous Adsorption Resin" (Chinese Journal of Experimental Formulas, Volume 16, No. 8, Page 22-24, July 2010) discloses a method for extracting total saponins of litchi seed, However, this method only extracts the single active ingredient of total saponins, and the obtained extract has limited effects. "Extraction process optimization of total saponins in litchi core" (Chinese Journal of Experimental Formulas, Volume 17, No. 12, Pages 48-49, June 2011) also discloses a method of ethanol extraction and ultrasonic assisted extraction for Extraction of total saponins in litchi core, but does not involve the extraction of fractions containing multiple active ingredients. CN 102048885A discloses a litchi nucleus saponin extraction process, which can extract the effective components of litchi nucleus, and the extracted litchi nucleus saponin content is as high as 85.0%, but it also does not mention the extraction of multiple effective fraction groups.

Contents of the invention

The object of the present invention is to provide a method for preparing the effective part group extract of litchi core with high extraction rate and simple process flow in view of the technical problems to be solved above.

In order to achieve the above object, the present invention provides a method for preparing the effective fraction group extract of litchi core, comprising the following steps:

(1) Take litchi core medicinal material, after crushing, add ethanol according to the ratio of material to liquid: 1g: 8-10ml, soak for 0.5-1 hour, perform ultrasonic extraction, and obtain extract A for later use;

(2) Add ethanol to the dregs according to the solid-liquid ratio of 1g: 8-10ml, soak for 0.5-1 hour, perform ultrasonic extraction to obtain extract B, and combine extract A and extract B;

(3) Reclaim ethanol from the combined extract A and extract B until there is no alcohol smell, then dilute the medicinal solution with distilled water to a concentration of 0.4 g/ml in terms of crude drug amount, leave it to stand overnight, filter, and the filtrate is for subsequent use;

(4) The filtrate passes through the AB-8 macroporous resin column, then first elutes with pure water of 3 times the amount of resin, discards the washing liquid, and then elutes the effective substance on the resin with ethanol of 5 times the amount of medicinal material, and collects ethanol eluent;

(5) reclaim the ethanol from the ethanol eluent collected in step (4) until there is no alcohol smell, and continue to concentrate into an extract;

(6) Evaporate the extract described in step (5) to dryness, and then carry out low-temperature vacuum drying to obtain the extract of effective fractions of litchi core.

As a preferred embodiment of the present invention, the concentration of ethanol used in the step (1), step (2) and step (4) is at least 70v/v%.

As a preferred embodiment of the present invention, the ethanol concentration used in the step (1) and step (2) is 70v/v%.

As a preferred embodiment of the present invention, the ethanol concentration used in the step (4) is 75v/v%.

As a preferred embodiment of the present invention, vacuum distillation is used to recover ethanol in the step (3) and step (5).

Further, the conditions of the vacuum distillation are vacuum degree -0.1 MPa and temperature 75°C.

As a preferred embodiment of the present invention, in the step (4), the flow velocity of the filtrate through the AB-8 macroporous resin column, the flow velocity of the elution with pure water, and the flow velocity of the elution with ethanol are 2 per hour. times the column bed volume.

As a preferred embodiment of the present invention, the relative density of the extract in the step (5) is 1.01-1.10.

As a preferred embodiment of the present invention, in the step (6), the extract is evaporated to dryness in a water bath.

Further, the temperature of drying in the water bath is 100°C

As a preferred embodiment of the present invention, in the step (6), the vacuum drying conditions are vacuum degree-0.1MPa, and drying at room temperature for 48 hours.

Through content determination, in the effective part group extract of litchi core obtained by the method of the present invention, the total flavonoids account for 20-30%, the total saponins account for 18-25%, and the tannins account for 25-35%. The sum of the contents of the three is 63-90%.

Compared with the prior art, the method of the present invention has the following obvious advantages: (1) the design of the process steps of the present invention is reasonable, the process flow is simple, and it is easy to operate; (2) the present invention adopts ethanol as the extraction solvent, combined with ultrasonic extraction, the extract Purified with AB-8 macroporous resin, the extract of effective parts containing total flavonoids, total saponins and tannins can be obtained at the same time, with clear ingredients and controllable quality, which meets the requirements of traditional Chinese medicine theory and modernization of traditional Chinese medicine; (3) effective parts The group extraction rate is high, and the extracted components are complete; (4) through recycling, ethanol and resin can be reused, which is economical and environmentally friendly, and can be effectively applied to industrial production.

Detailed ways

The present invention will be described in further detail below in conjunction with specific examples, but the present invention is not limited to the following examples.

Hereinafter, "5 times the amount of medicinal material" means that the ratio of the volume of ethanol used to the weight of crude drug added is 5ml:1g. For example, if you take 100g of lychee stone powder, you need to use 500ml of ethanol to elute.

Example 1

Crush the lychee pits into a coarse powder. Take 100g of lychee seed powder, add 1000ml of 70v/v% ethanol for the first time, soak for 0.5 hours, then ultrasonically extract for 1 hour, pour the extract A into the container for later use. Continue to add 800ml of 70v/v% ethanol to the medicinal residues, after soaking for 1 hour, ultrasonically extract for 1 hour, pour out the extract B, and combine it with the extract A obtained by the first extraction. The combined extract A and extract B are recovered to ethanol until there is no alcohol smell. The recovery method is preferably vacuum distillation recovery (vacuum degree -0.1MPa, temperature 75°C). The medicinal solution after recovery of ethanol was diluted with distilled water to 250ml, left standing overnight, filtered, and the filtrate was set aside. The filtrate first passes through the treated AB-8 macroporous resin column at a flow rate of 2 times the column bed volume per hour (see below for the treatment method), and then uses 3 times the resin amount of pure water at a flow rate of 2 times the column bed volume per hour. Elution, discard the washing liquid, then carry out the effective substance on the eluting resin with 75v/v% ethanol of 5 times of medicinal material amount (being 500ml) with the flow rate of 2 times of column bed volume per hour, and collect this ethanol elution liquid. The ethanol eluate is decompressed to recover ethanol until it has no alcohol smell (vacuum degree -0.1MPa, temperature 75°C), and then continue to concentrate to an extract with a relative density of 1.05 (the relative density refers to the density relative to water). Evaporate the extract to dryness in a water bath (100°C), put it into a vacuum drying oven for low-temperature vacuum drying (vacuum degree -0.1MPa, dry at room temperature for 48 hours), and obtain the extract of effective fractions of litchi core.

Among them, the processing method of AB-8 macroporous resin is as follows: Take AB-8 macroporous resin of clean pharmaceutical grade, put it into the resin column by wet method, remove the resin floating on the water surface, drain the water from the lower end, and use 95v Rinse the column layer with /v% ethanol until the eluent and water are mixed at a volume ratio of 1:1 and no turbidity appears, then rinse the column layer with a large amount of pure water until the effluent has no alcohol smell.

The extract of the effective part group of litchi core obtained above is measured by ultraviolet-visible spectrophotometry to know the content of relevant components wherein: the total flavonoid content (referring to the mass ratio of total solids) is about 21%, and the total saponin content (referring to the total solid content) is about 21%. The mass ratio of solids) is about 20%, and the tannin content (referring to the mass ratio of total solids) is about 30%.

Example 2

Crush the lychee pits into a coarse powder. Take 100g of lychee seed powder, add 800ml of 70v/v% ethanol for the first time, soak for 1 hour, ultrasonically extract for 1 hour, and pour the extract A into the container for later use. Continue to add 1000ml of 70v/v% ethanol to the dregs, soak for 0.5 hours, ultrasonically extract for 1 hour, pour out the extract B, and combine it with the extract A obtained by the first extraction. The combined extract A and extract B are recovered to ethanol until there is no alcohol smell. The recovery method is preferably vacuum distillation recovery (vacuum degree -0.1MPa, temperature 75°C). The medicinal solution after recovery of ethanol was diluted with distilled water to 250ml, left standing overnight, filtered, and the filtrate was set aside. The filtrate first passed through the treated AB-8 macroporous resin column at a flow rate of 2 times the column bed volume per hour, then eluted with 3 times the resin amount of pure water at a flow rate of 2 times the column bed volume per hour, and discarded the water for washing Liquid, then carry out eluting effective substances on the resin with 75v/v% ethanol of 5 times of medicinal material amount (ie 500ml) at a flow rate of 2 times of column bed volume per hour, and collect this ethanol eluent. The ethanol eluate is decompressed to recover ethanol until it has no alcohol smell (vacuum degree -0.1MPa, temperature 75°C), and continues to concentrate to an extract with a relative density of 1.01 to 1.10 (the relative density refers to the density relative to water. ). Evaporate the extract to dryness in a water bath (100°C), put it into a vacuum drying oven for low-temperature vacuum drying (vacuum degree -0.1MPa, dry at room temperature for 48 hours), and obtain the extract of effective fractions of litchi core.

Among them, the processing method of AB-8 macroporous resin is as follows: Take AB-8 macroporous resin of clean pharmaceutical grade, put it into the resin column by wet method, remove the resin floating on the water surface, drain the water from the lower end, and use 95v Rinse the column layer with /v% ethanol until the eluent and water are mixed at a volume ratio of 1:1 and no turbidity appears, then rinse the column layer with a large amount of pure water until the effluent has no alcohol smell.

The extract of the effective parts of litchi core obtained above is measured by ultraviolet-visible spectrophotometry and the content of related components can be known: the total flavonoid content (referring to the mass ratio of the total solids) is about 20%, and the total saponin content (referring to the total solid content) is about 20%. The mass ratio of solids) is about 25%, and the tannin content (referring to the mass ratio of total solids) is about 32%.

Example 3 Pilot scale-up test

Weigh 10Kg of lychee kernel coarse powder, add 100L of 70v/v% ethanol for the first time, soak for 0.5 hours, then ultrasonically extract for 1 hour, and pour the extract into a container for later use. Continue to add 80L of 70v/v% ethanol to the dregs, soak for 0.5 hours, ultrasonically extract for 1 hour, pour out the extract and combine it with the first extract; depressurize the combined total extract to recover ethanol until it has no alcohol smell (vacuum degree -0.1MPa, temperature 75°C), the extract was diluted with distilled water to a volume of 25L, left to stand overnight, filtered, and the filtrate was set aside. The filtrate first passed through the treated AB-8 macroporous resin column at a flow rate of 2 times the column bed volume per hour, then eluted with 3 times the resin amount of pure water at a flow rate of 2 times the column bed volume per hour, and discarded the water for washing liquid, then elute the effective substance on the resin with 75v/v% ethanol of 5 times the amount of medicinal material (ie 50L) at a flow rate of 2 times the column bed volume per hour, and collect this part of the ethanol eluate. Recover the ethanol eluate under reduced pressure until there is no alcohol smell (vacuum degree -0.1MPa, temperature 75°C), and continue to concentrate to an extract with a relative density of 1.01-1.10 (the relative density refers to the density relative to water) . Evaporate the above extract to dryness in a water bath (100°C), put it into a vacuum drying oven for low-temperature vacuum drying (vacuum degree -0.1MPa, dry at room temperature for 48 hours), and obtain the extract of effective fractions of litchi core.

Among them, the processing method of the macroporous resin is as follows: Take 20Kg of clean pharmaceutical grade AB-8 macroporous resin, put it into the resin column by wet method, remove the resin floating on the water surface, drain the water from the lower end, and use 95v/v Wash the column layer with % ethanol until the 1:1 mixture of the eluent and water does not appear turbid, and then rinse the column layer with a large amount of pure water until the effluent has no alcohol smell.

The obtained litchi core effective fraction group extract is measured through ultraviolet-visible spectrophotometry the content of active ingredient wherein, weighs the extract dry ointment quality, calculates ointment yield (extract ointment rate=extract dry ointment quality/medicinal material feeding quality × 100%)

Using the above process, a total of 3 batches of litchi seed extracts were prepared, and the results are shown in Table 1.

Table 1 Pilot test results

The results showed that the extraction rate of the extract was 3%, and the purification effect had been achieved; among the measurable components of the extract, the total flavonoid content reached 20.1-20.5%, the total saponin content reached 19.3-20%, and the tannin content reached 31.8-32.2%. %, the sum of the three is more than 71%, which meets the requirements of stable and controllable quality, meets the requirements of modernization of traditional Chinese medicine, has stable process, and can realize industrialized production.

Claims (1)

1. a preparation method of litchi nucleus effective fraction group extract, is characterized in that being made up of following steps: 1) Take lychee core medicinal material, crush and add ethanol according to the material-to-liquid ratio of 1g: 8-10ml, soak for 0.5-1 hour, and perform ultrasonic extraction to obtain extract A for later use; 2) Add ethanol to the medicinal dregs according to the ratio of material to liquid: 1 g: 8-10 ml, soak for 0.5-1 hour, perform ultrasonic extraction to obtain extract B, and combine extract A and extract B; 3) Recover ethanol from the combined extract A and extract B until there is no alcohol smell, then dilute the medicinal liquid with distilled water to a concentration of 0.4 g/ml in terms of crude drug amount, let it stand overnight, filter, and use the filtrate for later use; 4) The filtrate passes through the AB-8 macroporous resin column, and then eluted with pure water 3 times the amount of the resin, discarded the washing solution, and then eluted the active substance on the resin with 5 times the amount of ethanol of the medicinal material, and collected the ethanol eluent; 5) Recover ethanol from the ethanol eluate collected in step 4) until it has no alcohol smell, and continue to concentrate it into an extract; 6) Evaporate the extract described in step 5) to dryness, and then carry out low-temperature vacuum drying to obtain the extract of effective parts of litchi core. 2. The preparation method according to claim 1, characterized in that: the concentration of ethanol used in step 1) and step 2) is 70v/v%. 3. The preparation method according to claim 1, characterized in that: the concentration of ethanol used in step 4) is 75 v/v%. 4. The preparation method according to claim 1, characterized in that the ethanol is recovered by vacuum distillation in the step 3) and step 5). 5. The preparation method according to claim 4, characterized in that: the conditions of the vacuum distillation are vacuum degree -0.1MPa and temperature 75°C. 6. The preparation method according to claim 1, characterized in that: in step 4), the flow rate of the filtrate passing through the AB-8 macroporous resin column, the flow rate of elution with pure water, and the flow rate of elution with ethanol Both are 2 times the column bed volume per hour. 7. The preparation method according to claim 1, characterized in that the relative density of the extract in the step 5) is 1.01-1.10. 8. The preparation method according to claim 1, characterized in that: in the step 6), the extract is evaporated to dryness in a water bath. 9. The preparation method according to claim 8, characterized in that: the temperature of the water bath for evaporating to dryness is 100°C. 10. The preparation method according to claim 1, characterized in that: in the step 6), the vacuum drying condition is a vacuum degree of -0.1 MPa, and drying at room temperature for 48 hours.