CN-122011080-A - A method for separating asiaticoside and madecassoside from herba Centellae

CN122011080ACN 122011080 ACN122011080 ACN 122011080ACN-122011080-A

Abstract

The invention relates to the technical field of natural product separation and purification, in particular to a preparation method for separating asiaticoside and madecassoside from centella, which comprises the steps of crushing centella and extracting with ethanol to obtain crude extract; removing fat-soluble impurities by adopting an aliphatic hydrocarbon organic solvent, extracting and enriching target glycosides by adopting a medium-polarity oxygen-containing organic solvent, determining a washing solvent by detecting physical and chemical parameters of an extract, washing the extract to obtain a purified extract, decoloring and concentrating to obtain centella asiatica total glycosides, obtaining a crystallization performance predicted value based on mass fingerprint data of the total glycosides, monitoring data in real time and dynamically adjusting the process in the crystallization process, and finally obtaining high-purity asiaticoside and madecassoside finished products respectively through fractional crystallization. The method has simple operation, recoverable solvent and obvious economical efficiency and mass production feasibility.

Inventors

LIU ZHIMOU
FENG LEI
YANG MIN
WANG HUIMING
YUE YAWU
WANG HAIMING
WEN JUPING

Assignees

湖南诺泽生物科技有限公司

Dates

Publication Date: 20260512
Application Date: 20260129

Claims (10)

1. A process for the preparation of asiaticoside and madecassoside isolated from centella asiatica, characterized in that it comprises the following steps: S1, reflux-extracting crushed centella asiatica medicinal materials with an ethanol solution to obtain a crude concentrated solution; s2, adding an aliphatic hydrocarbon organic solvent into the crude extract concentrate, extracting and retaining a water phase, adding a medium-polarity oxygen-containing organic solvent, and extracting to obtain an extract rich in asiaticoside and madecassoside; S3, determining a washing solvent based on physical and chemical parameters of the extract, and washing the extract to obtain a purified extract; S4, decoloring, concentrating and drying the purified extract to obtain centella asiatica total glycosides; S5, determining recrystallization times based on the predicted crystallization data of the asiaticoside and the actually detected crystallization monitoring data of the asiaticoside crude product output by the crystallization performance prediction model; S6, drying the asiaticoside crude product to obtain an asiaticoside finished product; S7, adding an alkyl acetate organic solvent into the crystallization mother liquor, crystallizing and separating to obtain a madecassoside crude product, and drying to obtain a madecassoside finished product.
2. The process for the isolation of asiaticoside and madecassoside from centella asiatica according to claim 1, characterized in that the aliphatic organic solvent comprises n-hexane or petroleum ether.
3. The method for preparing asiaticoside and madecassoside separated from centella asiatica according to claim 1, wherein the medium-polarity oxygen-containing organic solvent comprises saturated n-butanol or ethyl acetate, and the alkyl acetate organic solvent comprises methyl acetate, ethyl acetate, propyl acetate or butyl acetate.
4. The process for the preparation of asiaticoside and madecassoside isolation from centella asiatica according to claim 1, characterized in that said physicochemical parameters comprise the pH value of the extracts enriched in asiaticoside and madecassoside and/or the uv absorbance value at 254-280 nm.
5. The method for preparing asiaticoside and madecassoside isolated from centella asiatica according to claim 1, wherein the washing solvent comprises at least one of aqueous salt solution, aqueous acid solution, aqueous alkali solution and aqueous purified solution; Wherein the acid aqueous solution comprises at least one of aqueous solution prepared from hydrochloric acid, sulfuric acid, phosphoric acid and acetic acid, and the alkali aqueous solution comprises at least one of aqueous solution prepared from sodium hydroxide, sodium bicarbonate and sodium carbonate.
6. The method for preparing asiaticoside and madecassoside isolated from centella asiatica according to claim 1, characterized in that the process for determining the washing solvent based on the detection of said physicochemical parameters comprises: respectively comparing the pH value and the pH threshold value in the physicochemical parameters with the ultraviolet absorbance value and the ultraviolet absorbance threshold value at 254-280nm wavelength; And determining that the washing solvent is an alkaline aqueous solution and a saline solution based on the pH value being less than a first pH threshold or the ultraviolet absorbance value at the wavelength of 254-280nm being greater than a first ultraviolet absorbance threshold.
7. The method for preparing asiaticoside and madecassoside isolated from centella asiatica according to claim 1, characterized in that the process for determining the washing solvent based on the detection of the physicochemical parameter further comprises: determining that the washing solvent is an alkaline aqueous solution or a salt aqueous solution based on a pH value of equal to or greater than a first pH threshold and equal to or less than a second pH threshold or a uv absorbance value at a wavelength of 254-280nm of equal to or less than a first uv absorbance threshold and equal to or greater than a second uv absorbance threshold; wherein the first pH threshold is less than the second pH threshold and the first ultraviolet absorbance threshold is greater than the second ultraviolet absorbance threshold.
8. The method for preparing asiaticoside and madecassoside isolated from centella asiatica according to claim 1, characterized in that the process for determining the washing solvent based on the detection of the physicochemical parameter further comprises: Determining that the washing solvent is a salt solution or a purified water solution based on the pH value being greater than or equal to a first pH threshold and less than or equal to a second pH threshold or the ultraviolet absorbance value at a wavelength of 254-280nm being less than a second ultraviolet absorbance threshold; wherein the first pH threshold is less than the second pH threshold and the first ultraviolet absorbance threshold is greater than the second ultraviolet absorbance threshold.
9. The method for preparing asiaticoside and madecassoside isolated from centella asiatica according to claim 1, characterized in that the process of the predicted crystallization data of the total asiaticoside based on the crystallization property prediction model comprises: obtaining chromatographic peak area ratio of asiaticoside and madecassoside, absorbance value of asiaticoside at 280nm and 420nm, and appearance color of asiaticoside as quality fingerprint data; Inputting the quality fingerprint data into a crystallization performance prediction model, wherein the output prediction crystallization data are prediction crystallization induction time, prediction crystal morphology and prediction purity value; The crystallization performance prediction model is a regression model trained based on historical quality fingerprint data.
10. The method for preparing asiaticoside and madecassoside isolated from centella asiatica according to claim 9, characterized in that the process for determining the number of recrystallisation comprises: Determining the crystal morphology of the asiaticoside crude product based on the crystallization monitoring data; determining crystallization process stability based on the crystal morphology from a deviation of crystallization induction time relative to the predicted crystallization induction time; Based on the crystallization process stability, the number of recrystallisations is determined from the purity difference of the purity value relative to the predicted purity value.

Description

A method for separating asiaticoside and madecassoside from herba Centellae Technical Field The invention relates to the technical field of natural product separation and purification, in particular to a preparation method for separating asiaticoside and madecassoside from centella asiatica. Background Asiaticoside and madecassoside are main bioactive triterpene saponin components in centella asiatica, have various pharmacological activities of promoting wound healing, anti-inflammatory and anti-depression, and are widely applied in the fields of medicines, cosmetics and health care products. CN106380504B discloses a method for extracting madecassoside from centella asiatica, which adopts macroporous adsorption resin method for separation, but is limited by adsorption capacity of resin, limited sample loading amount, long separation period, large solvent consumption, and challenges in efficiency and cost in mass production. Chinese patent publication No. CN119431486A discloses a method for obtaining high-content madecassoside by centella asiatica by using a membrane separation method, which adopts a membrane separation technology to effectively remove impurities so as to improve the purity of madecassoside, but only madecassoside is obtained, and the systematic separation of two components is not successfully realized. The invention uses a solvent method, but does not completely separate two components of asiaticoside and madecassoside, and the used halohydrocarbon solvent has higher toxicity, does not accord with the green chemical industry trend, and cannot be industrialized. Therefore, the development of the separation and purification method which is low in cost and easy to industrialize and can simultaneously and efficiently obtain two high-purity monomers of asiaticoside and madecassoside has urgent demands and important significance for the industrialized application of asiaticoside components. Disclosure of Invention Therefore, the invention provides a preparation method for separating asiaticoside and madecassoside from centella asiatica, which is used for solving the problems of low separation efficiency of two components of asiaticoside and madecassoside, low purity of finished products, small treatment capacity and difficulty in large scale in the prior art. In order to achieve the above object, the present invention provides a method for preparing asiaticoside and madecassoside from centella asiatica, comprising the steps of: S1, reflux-extracting crushed centella asiatica medicinal materials with an ethanol solution to obtain a crude concentrated solution; s2, adding an aliphatic hydrocarbon organic solvent into the crude extract concentrate, extracting and retaining a water phase, adding a medium-polarity oxygen-containing organic solvent, and extracting to obtain an extract rich in asiaticoside and madecassoside; S3, determining a washing solvent based on physical and chemical parameters of the extract, and washing the extract to obtain a purified extract; S4, decoloring, concentrating and drying the purified extract to obtain centella asiatica total glycosides; S5, determining recrystallization times based on the predicted crystallization data of the asiaticoside and the actually detected crystallization monitoring data of the asiaticoside crude product output by the crystallization performance prediction model; S6, drying the asiaticoside crude product to obtain an asiaticoside finished product; S7, adding an alkyl acetate organic solvent into the crystallization mother liquor, crystallizing and separating to obtain a madecassoside crude product, and drying to obtain a madecassoside finished product. Further, the aliphatic hydrocarbon organic solvent includes n-hexane or petroleum ether. Further, the medium-polarity oxygen-containing organic solvent comprises saturated n-butanol or ethyl acetate, and the alkyl acetate organic solvent comprises methyl acetate, ethyl acetate, propyl acetate or butyl acetate. Further, the physicochemical parameters include the pH value of the extract enriched in asiaticoside and madecassoside and/or the UV absorbance value at 254-280 nm. Further, the washing solvent includes at least one of an aqueous salt solution, an aqueous acid solution, an aqueous alkali solution, and an aqueous purified solution; Wherein the acid aqueous solution comprises at least one of aqueous solution prepared from hydrochloric acid, sulfuric acid, phosphoric acid and acetic acid, and the alkali aqueous solution comprises at least one of aqueous solution prepared from sodium hydroxide, sodium bicarbonate and sodium carbonate. Further, the process of determining the washing solvent based on the detection of the physicochemical parameter comprises: respectively comparing the pH value and the pH threshold value in the physicochemical parameters with the ultraviolet absorbance value and the ultraviolet absorbance threshold value at 254-280nm wavelength; And determining that the washing