METHOD FOR EXTRACTING ASTRAGALOSIDE IV FROM FRESH RADIX ASTRAGALI

Abstract

The disclosure discloses a method for extracting Astragaloside IV from fresh Radix Astragali. The method comprises the following steps: cleaning and cutting fresh Radix Astragali roots into sections, and extracting with an ethanol solution for multiple times under an ultrasonic condition; merging the extracting solutions, concentrating to remove ethanol, hydrolyzing the concentrate with sodium hydroxide-calcium hydroxide mixed alkali, filtering the precipitate; then flocculating and precipitating by using a flocculating agent chitosan, and filtering; removing impurities through a flocculation-precipitation mode and filtering with a filter screen; separating through a macroporous adsorption resin, and finally purifying through recrystallization to obtain a high-purity Astragaloside IV. The method has the advantages of easily available raw materials, low cost, high Astragaloside IV extraction efficiency, high Astragaloside IV purity and simple process, the raw materials involved in the extraction process are non-toxic and harmless, and it is suitable for large-scale industrial production.

Claims

1. A method for extracting Astragaloside IV from fresh Radix Astragali, comprising the following steps: (a) Cleaning the fresh Radix Astragali roots, and cutting the roots into sections; (b) Under ultrasonic conditions, extracting several times with an ethanol solution to obtain an extract, concentrating under reduced pressure to remove ethanol to obtain a concentrate, adding water to the concentrate to 4-8 times the weight of the fresh Radix Astragali, and filtering to obtain a concentrated solution; (c) Adjusting the pH of the concentrated solution to maintain at 11-13 with sodium hydroxide-calcium hydroxide mixed alkali under a condition of 40-60° C., keeping temperature and hydrolyzing for 2-4 hours, filtering with a filter screen, and collecting a filtrate; (d) Adjusting a pH of the filtrate with an acid to 5-7, then adding a chitosan solution for flocculation, filtering with a filter screen, and collecting a filtrate; (e) Separating the obtained filtrate through a macroporous adsorption resin column to obtain a crude Astragaloside IV; and (f) Recrystallizing the crude Astragaloside IV with methanol or ethanol to obtain a high-purity Astragaloside IV.

2. The method for extracting Astragaloside IV from fresh Radix Astragali according to claim 1, wherein the cutting into sections in the step (a) is cutting into small sections with a length of 3-5 mm.

3. The method for extracting Astragaloside IV from fresh Radix Astragali according to claim 1, wherein the ultrasonic condition in the step (b) include ultrasonic power between 300-350 W, temperature between 40-60° C., and extraction time between 30-60 minutes.

4. The method for extracting Astragaloside IV from fresh Radix Astragali according to claim 1, wherein the ethanol solution in step (b) is a 70-85% ethanol solution, the extracting for several times is extracting for 3-4 times, and the amount of ethanol solution used for each extraction is 4-10 times the weight of fresh Radix Astragali.

5. The method for extracting Astragaloside IV from fresh Radix Astragali according to claim 1, wherein a weight ratio of sodium hydroxide to calcium hydroxide in the mixed alkali in the step (c) is 1:2-10.

6. The method for extracting Astragaloside IV from fresh Radix Astragali according to claim 1, wherein the chitosan solution in the step (d) is a 1% chitosan solution prepared with 1% acetic acid aqueous solution as a solvent, and the amount of chitosan in the filtrate is 0.1-0.5%.

7. The method for extracting Astragaloside IV from fresh Radix Astragali according to claim 1, wherein the filter screens used in the step (c) and step (d) are made of stainless steel, with a pore size of 200-500 mesh.

8. The method for extracting Astragaloside IV from fresh Radix Astragali according to claim 1, wherein the acid in step (d) is dilute hydrochloric acid or dilute sulfuric acid.

9. The method for extracting Astragaloside IV from fresh Radix Astragali according to claim 1, wherein the macroporous adsorption resin in the step (e) is AB-8, D101 or HPD100.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0018] FIG. 1 is a 1H NMR (DMSO-d6) spectra of Astragaloside IV

DETAILED DESCRIPTION OF THE EMBODIMENTS

[0019] The technical solution of the present disclosure is further described in detail below through specific embodiments. The sodium hydroxide-calcium hydroxide mixed alkali solution used in the examples is prepared with a mass ratio of 1:6-8; the chitosan solution is a 1% chitosan solution prepared with 1% acetic acid aqueous solution as the solvent. An ultrasonic condition used for extractions is defined by using an ultrasonic extraction device having variable settings for an ultrasonic power, a temperature range, and a length of time for an extraction.

Example 1

[0020] In a 5 L ultrasonic extraction device, 1 kg of fresh Radix Astragali roots that were cleaned and cut into sections were added, 5.0 L of 80% ethanol solution was added, the ultrasonic power was adjusted to 350 W, the extraction temperature was 55° C., the extract was carried out for 40 minutes and 3 times, the extracts were combined, the ethanol was removed under reduced pressure to obtain a concentrate; 2.0 L of water was added to the concentrate to obtain a concentrated solution, and the concentrated solution was filtered. The sodium hydroxide-calcium hydroxide (mass ratio of 1:8) mixed alkali was added to the filtrate, the pH was adjusted to 13, stirred at 45° C. for 3.5 hours, a resulting solution was allowed to stand, and filtered with a 250 mesh filter screen; the pH of filtrate was adjusted to 5 with dilute hydrochloric acid, under stirring at 40° C., 0.6 L of chitosan solution was added to the filtrate, left to stand, and filtered with a 500 mesh stainless steel filter screen. The filtrate was adsorbed on AB-8 macroporous adsorption resin column and eluted with 80% ethanol solution, the eluent was concentrated to a small volume, and the precipitate appeared on standing, the precipitate was obtained by centrifugation. Finally, it was recrystallized with methanol to obtain 0.40 g of purified Astragaloside IV product with a purity of 97.5%, the 1H NMR (DMSO-d6) spectra of Astragaloside IV is shown in FIG. 1.

Example 2

[0021] In a 5 L ultrasonic extraction device, 1 kg of fresh Radix Astragali roots that were cleaned and cut into sections were added, 6.0 L of 75% ethanol solution was added, the ultrasonic power was adjusted to 300 W, the extraction temperature was 60° C., the extract was carried out for 60 minutes and 3 times, the extracts were combined, the ethanol was removed under reduced pressure to obtain a concentrate; 0.5 L of water was added to the concentrate to obtain a concentrated solution, and the concentrated solution was filtered. The sodium hydroxide-calcium hydroxide (mass ratio of 1:8) mixed alkali was added to the filtrate, the pH was adjusted to 13, stirred at 50° C. for 4 hours, a resulting solution was allowed to stand, and filtered with a 200 mesh filter screen; the pH of filtrate was adjusted to 6 with dilute hydrochloric acid, under stirring at 45° C., 0.8 L of chitosan solution was added to the filtrate, left to stand, and filtered with a 500 mesh stainless steel filter screen. The filtrate was adsorbed on D101 macroporous adsorption resin column and eluted with 80% ethanol solution, the eluent was concentrated to a small volume, and the precipitate appeared on standing, the precipitate was obtained by centrifugation. Finally, it was recrystallized with methanol to obtain 0.35 g of purified Astragaloside IV product with a purity of 98.0%.

Example 3

[0022] In a 5 L ultrasonic extraction device, 1 kg of fresh Radix Astragali roots that were cleaned and cut into sections were added, 8.0 L of 85% ethanol solution was added, the ultrasonic power was adjusted to 350 W, the extraction temperature was 55° C., the extract was carried out for 40 minutes and 3 times, the extracts were combined, the ethanol was removed under reduced pressure to obtain a concentrate; 1.5 L of water was added to the concentrate to make a concentrated solution, and the concentrated solution was filtered. The sodium hydroxide-calcium hydroxide (mass ratio of 1:6) mixed alkali was added to the filtrate, the pH was adjusted to 12, stirred at 40° C. for 4 hours, a resulting solution was allowed to stand, and filtered with a 200 mesh filter screen; the pH of filtrate was adjusted to 5 with dilute hydrochloric acid, under stirring at 40° C., 1.0 L of chitosan solution was added to the filtrate, left to stand, and filtered with a 300 mesh stainless steel filter screen. The filtrate was adsorbed on HPD-100 macroporous adsorption resin column and eluted with 80% ethanol solution, the eluent was concentrated to a small volume, and the precipitate appeared on standing, the precipitate was obtained by centrifugation. Finally, it was recrystallized with alcohol to obtain 0.42 g of purified Astragaloside IV product with a purity of 97.0%.