CN113831385A - Method for extracting tenuifolin from polygala tenuifolia - Google Patents

Abstract

The invention belongs to the field of traditional Chinese medicine extraction, and particularly discloses a rapid and efficient preparation method of tenuifolin, which comprises the following specific implementation steps: decocting polygala tenuifolia medicinal materials with water for extraction, concentrating an extracting solution, then carrying out alkaline hydrolysis, adjusting the pH value to 4-5 with acid, adding absolute ethyl alcohol for alcohol precipitation, concentrating an alcohol precipitation solution until no alcohol smell exists, carrying out water precipitation, centrifuging, collecting precipitate, drying to obtain crude product of leaf polygala tenuifolia saponin, and repeatedly recrystallizing the obtained crude product to obtain the leaf polygala tenuifolia saponin with the content of more than 90%. The method has the advantages of low cost, short production period, green process, low environmental pollution, simple operation and high yield, and is suitable for large-scale industrial production.

Description

Method for extracting tenuifolin from polygala tenuifolia

Technical Field

The invention belongs to the field of traditional Chinese medicine extraction, and particularly relates to a method for extracting tenuifolin from polygala tenuifolia.

Background

Polygala tenuifolia, also known as \33917, Tokyo, 34112, \33964m. Producing northeast, northwest, central and Sichuan; perennial herbs, strong and strong taproots, and phloem flesh. Has effects in tranquilizing mind, improving intelligence, eliminating phlegm, and relieving swelling, and can be used for treating insomnia, dreaminess, amnesia, palpitation, absentmindedness, expectoration, skin sore, swelling and toxin, and breast swelling and pain caused by imbalance between heart-yang and kidney-yin. According to the report of the literature, polygala tenuifolia saponin has strong hemolytic effect and high toxicity, and researches show that ester bonds at 28 th position of mother nucleus are broken after polygala tenuifolia saponin is subjected to alkaline hydrolysis, and are converted into secondary saponin tenuifolin (CAS 20183-47-5), the hemolytic effect disappears after hydrolysis, and the toxicity is remarkably reduced. Pharmacological experiments show that the tenuifolin has the capability of remarkably improving the learning and memory of mice in aging models, has potential application prospects in the aspect of preventing and treating senile dementia, and attracts people's attention.

The tenuifolin has stable chemical property and molecular formula C₃₆H₅₆O₁₂Molecular weight 680.37, structural formula:

in the prior art, the extraction process of the tenuifolin is mostly fussy, the types of the existing procedures are less, the variability is lower, and some simplified, replaced and novel steps are also arranged in the procedures. For example, 201210010622.3 patent and 201110074847.0 patent adopt silica gel column chromatography and polyamide column chromatography to separate tenuifolin; 201810774023.6 patent comprises extracting with methanol, separating with macroporous adsorbent resin and silica gel column chromatography to obtain tenuifolin; in patent 201811002097.4, tenuifolin is obtained by extracting with 90% ethanol and then performing subsequent treatment.

Disclosure of Invention

In order to overcome the defects of the preparation process of tenuifolin in the prior art, the invention provides the method for extracting the tenuifolin from polygala tenuifolia, which has the advantages of simple process, low cost, environmental protection and suitability for industrial mass production.

The main purpose of the invention is realized by the following technical scheme:

a method for preparing tenuifolin mainly comprises the following steps:

(1) decocting and extracting with water: weighing polygala tenuifolia, extracting with drinking water to obtain water extract, and concentrating the water extract to obtain concentrated solution;

(2) alkaline hydrolysis: adding alkali into the concentrated solution obtained in the step (1) for alkaline hydrolysis to obtain alkaline hydrolysis solution;

(3) alcohol precipitation: adjusting the pH value of the alkaline hydrolysis liquid obtained in the step (2) to 4-5 by using acid, adjusting the alcohol degree to 70% -90% by using absolute ethyl alcohol, filtering, and collecting filtrate;

(4) water precipitation: concentrating the filtrate collected in the step (3) until no alcohol smell exists, adding drinking water for water precipitation, centrifuging to collect precipitate, and drying to obtain crude tenuifolin;

(5) and (3) crystallization: recrystallizing the crude tenuifolin obtained in the step (4) to obtain the tenuifolin.

Preferably, the drinking water is added in the step (1) for extraction for 1-3 times; the extraction time is 1-4h each time; the ratio (m/V) of the drinking water (V) to the medicinal material (m) is 1:10-1: 20.

Preferably, the alkaline hydrolysis concentration in the step (2) is 1-10%, and the alkaline hydrolysis time is 1-3 h;

in the step (3), the acid is phosphoric acid, hydrochloric acid or acetic acid;

the amount of the drinking water added in the step (4) is as follows: adding 25L-40L of medicinal materials per kilogram;

the crystallization solvent in the step (5) is: absolute ethanol or methanol.

A method for preparing tenuifolin mainly comprises the following steps:

(1) decocting and extracting with water: weighing cortex et radix Polygalae, adding 15 times of drinking water, extracting for 3 times (4 hr each time), mixing water extractive solutions, and concentrating to obtain concentrated solution;

(2) alkaline hydrolysis: adding alkali into the concentrated solution obtained in the step (1) until the concentration is 10%, and carrying out alkaline hydrolysis for 3h to obtain alkaline hydrolysis solution;

(3) alcohol precipitation: adjusting the pH value of the alkaline hydrolysis liquid obtained in the step (2) to 4-5 by using concentrated hydrochloric acid, adjusting the alcohol degree to 70% -90% by using absolute ethyl alcohol, filtering, and collecting filtrate;

(4) water precipitation: concentrating the filtrate collected in the step (3) until no alcohol smell exists, adding 40L of drinking water into each kilogram of medicinal materials for water precipitation, centrifugally collecting precipitate, and drying to obtain crude tenuifolin;

(5) and (3) crystallization: and (4) carrying out reflux recrystallization on the crude tenuifolin obtained in the step (4) by using absolute ethyl alcohol to obtain the tenuifolin.

Compared with the prior art, the preparation method of tenuifolin provided by the invention has the following advantages:

(1) the invention takes the green organic solvent as the extraction solvent, and the environmental protection pressure is small.

(2) The steps of separation and purification are reduced, so that the preparation method of the tenuifolin is simpler and more effective, the cost is low, the yield is high, the requirement on production equipment is low, and the method is more suitable for large-scale industrial production.

DETAILED DESCRIPTION OF EMBODIMENT (S) OF INVENTION

The present invention will be further described with reference to specific embodiments, but the scope of the invention as claimed is not limited to the following embodiments.

Example 1

500.3g of polygala tenuifolia medicinal material is added with 7.5L of drinking water, heating reflux extraction is carried out for 4h, extraction is carried out for 3 times, extracting solutions are merged and concentrated into clear paste, caustic soda flakes are added and adjusted to 10%, alkaline hydrolysis is carried out for 3h, concentrated hydrochloric acid is used for adjusting the pH value to 4.40, absolute ethyl alcohol is added for adjusting the alcohol degree to 80%, filtration is carried out, filtrate is collected, the filtrate is concentrated to be free of alcohol smell (the alcohol degree is 0), 20.0L of drinking water is added for water precipitation, precipitate is centrifugally collected, drying is carried out, 8.20g of tenuifolin crude product is obtained, absolute ethyl alcohol is added for repeated reflux recrystallization, 1.35g of tenuifolin is obtained, the content is 92.6%, and the yield is 10.20%.

Example 2

3600.4g of polygala tenuifolia medicinal material is added with 54.0L of drinking water, heating reflux extraction is carried out for 2 hours, extraction is carried out for 2 times, extracting solutions are merged and concentrated into clear paste, caustic soda flakes are added and adjusted to 5%, alkaline hydrolysis is carried out for 2 hours, pH is adjusted to 4.85 by phosphoric acid, absolute ethyl alcohol is added to adjust the alcohol degree to 70%, filtration is carried out, filtrate is collected, the filtrate is concentrated to be free of alcohol smell (the alcohol degree is 0), 90.0L of drinking water is added for water precipitation, precipitate is centrifugally collected, drying is carried out, crude tenuifolin is obtained, 36.10g of tenuifolin is obtained, methanol is added for repeated reflux recrystallization, 9.30g of tenuifolin is obtained, the content is 92.1%, and the yield is 9.70%.

Example 3

6400.2g of polygala tenuifolia medicinal material is added with 64.0L of drinking water to be heated and refluxed for extraction for 1 hour, the mixture is extracted for 1 time, the extracting solution is condensed into clear paste, caustic soda flakes are added to be adjusted to 10 percent, alkaline hydrolysis is carried out for 1 hour, the pH value is adjusted to 4.30 by acetic acid, absolute ethyl alcohol is added to adjust the alcohol degree to 80 percent, filtration is carried out, filtrate is collected, the filtrate is concentrated to be free of alcohol smell (the alcohol degree is 0), about 160.0L of drinking water is added for water precipitation, the precipitate is centrifugally collected, the precipitate is dried, 92.20g of crude tenuifolin is obtained, absolute ethyl alcohol is added to carry out repeated reflux recrystallization to obtain 19.10g of tenuifolin, the content is 94.20 percent, and the yield is 9.50 percent.

Example 4

500.6g of polygala tenuifolia medicinal material is added with 10.0L of drinking water, heating reflux extraction is carried out for 2 hours, extraction is carried out for 2 times, extracting solutions are merged and concentrated into clear paste, caustic soda flakes are added, the solution is adjusted to 1 percent and alkaline hydrolysis is carried out for 3 hours, the pH value is adjusted to 4.0 by concentrated hydrochloric acid, absolute ethyl alcohol is added to adjust the alcohol degree to 90 percent, filtration is carried out, filtrate is collected, the filtrate is concentrated to be free of alcohol smell (the alcohol degree is 0), 12.5L of drinking water is added for water precipitation, a precipitate part is centrifugally collected, drying is carried out, 8.10g of crude tenuifolin is obtained, absolute ethyl alcohol is added for repeated reflux recrystallization, 1.05g of tenuifolin is obtained, the content is 90.6 percent, and the yield is 7.76 percent.

Example 5

500.8g of polygala tenuifolia medicinal material is added with 10.0L of drinking water, heating reflux extraction is carried out for 2 hours, extraction is carried out for 2 times, extracting solutions are merged and concentrated into clear paste, caustic soda flakes are added and adjusted to 5%, alkaline hydrolysis is carried out for 2 hours, concentrated hydrochloric acid is used for adjusting pH to 5.0, absolute ethyl alcohol is added for adjusting alcohol degree to 80%, filtration is carried out, filtrate is collected, the filtrate is concentrated until the alcohol degree is 0 without alcohol taste, 15.0L of drinking water is added for carrying out water sedimentation, a sediment part is centrifugally collected, drying is carried out, a crude product of tenuifolin is obtained, 8.00g of tenuifolin is obtained, methanol is added for repeated reflux recrystallization, 1.19g of tenuifolin is obtained, the content is 91.6%, and the yield is 8.89%.

Claims (8)

1. A method for preparing tenuifolin mainly comprises the following steps

(1) Decocting and extracting with water: weighing polygala tenuifolia, adding drinking water for extraction to obtain water extract, and concentrating the water extract to obtain concentrated solution;

(2) alkaline hydrolysis: adding alkali into the concentrated solution obtained in the step (1) for alkaline hydrolysis to obtain alkaline hydrolysis solution;

(3) alcohol precipitation: adjusting the pH value of the alkaline hydrolysis liquid obtained in the step (2) to 4-5 by using acid, adjusting the alcohol degree to 70% -90% by using absolute ethyl alcohol, filtering, and collecting filtrate;

(4) water precipitation: concentrating the filtrate collected in the step (3) until no alcohol smell exists, adding drinking water for water precipitation, centrifuging to collect precipitate, and drying to obtain crude tenuifolin;

(5) and (3) crystallization: recrystallizing the crude tenuifolin obtained in the step (4) to obtain the tenuifolin.

2. The method according to claim 1, wherein the drinking water added in the step (1) is extracted for 1 to 3 times; the extraction time is 1-4 hr.

3. The method of claim 1, wherein: the ratio (m/V) of the drinking water (V) to the medicinal material (m) used in the step (1) is 1:10-1: 20.

4. The method of claim 1, wherein: the alkaline hydrolysis concentration in the step (2) is 1-10%, and the alkaline hydrolysis time is 1-3 h.

5. The method of claim 1, wherein: in the step (3), the acid is phosphoric acid, hydrochloric acid or acetic acid.

6. The method of claim 1, wherein the amount of drinking water added in step (4) is 25L-40L per kg of the crude drug.

7. The method of claim 1, wherein the crystallization solvent in step (5) is: absolute ethanol or methanol.

8. A method for preparing tenuifolin mainly comprises the following steps:

(1) decocting and extracting with water: weighing cortex et radix Polygalae, adding 10 times of drinking water, extracting for 3 times (4 hr each time), mixing water extractive solutions, and concentrating to obtain concentrated solution;

(2) alkaline hydrolysis: adding alkali into the concentrated solution obtained in the step (1) until the concentration is 10%, and carrying out alkaline hydrolysis for 3h to obtain alkaline hydrolysis solution;

(3) alcohol precipitation: adjusting the pH value of the alkaline hydrolysis liquid obtained in the step (2) to 4-5 by using concentrated hydrochloric acid, adjusting the alcohol degree to 70% -90% by using absolute ethyl alcohol, filtering, and collecting filtrate;

(4) water precipitation: concentrating the filtrate collected in the step (3) until no alcohol smell exists, adding 40L of drinking water into each kilogram of medicinal materials for water precipitation, centrifugally collecting precipitate, and drying to obtain crude tenuifolin;

(5) and (3) crystallization: and (4) carrying out reflux recrystallization on the crude tenuifolin obtained in the step (4) by using absolute ethyl alcohol to obtain the tenuifolin.