阅读说明：本技术 基于提取分离法测定大蓟炭化学成分的方法 (Method for determining chemical components of thistle charcoal based on extraction and separation method ) 是由 龚千锋 于欢 严丽萍 于 2019-11-12 设计创作，主要内容包括：本发明公开了基于提取分离法测定大蓟炭化学成分的方法,包括以下步骤：S1、制备浸膏：大蓟炭5kg,粉碎后70%乙醇回流提取3次,合并滤液减压浓缩,得浸膏800g；S2、浸膏分离；取浸膏800g溶解于8L蒸馏水中,依次用石油醚、氯仿、乙酸乙酯分别萃取3次,回收溶剂后得石油醚部分87g、氯仿部分93g、乙酸乙酯部分56g、氯仿萃取部分90g；S3、洗脱合并：将浸膏分离获取成分经硅胶柱层析氯仿、甲醇梯度洗脱合并为8个部分,计为Fr1-Fr6。本发明通过大孔吸附树脂、正相硅胶柱色谱手段进行分离,通过对波谱学数据分析,确定化合物的成分,实现为大蓟和成分研究提供相关参考依据。(The invention discloses a method for determining chemical components of thistle carbon based on an extraction and separation method, which comprises the following steps: s1, preparing an extract: pulverizing 5kg of thistle charcoal, extracting with 70% ethanol under reflux for 3 times, mixing filtrates, and concentrating under reduced pressure to obtain 800g of extract; s2, separating the extract; dissolving 800g of the extract in 8L of distilled water, sequentially extracting with petroleum ether, chloroform and ethyl acetate for 3 times respectively, and recovering the solvent to obtain 87g of petroleum ether part, 93g of chloroform part, 56g of ethyl acetate part and 90g of chloroform extraction part; s3, elution and combination: separating the extract to obtain components, subjecting to silica gel column chromatography, gradient eluting with chloroform and methanol, and mixing into 8 parts, which are counted as Fr1-Fr 6. The method separates by means of macroporous adsorption resin and normal phase silica gel column chromatography, and determines the components of the compound by analyzing the data of the spectroscopy, thereby realizing the purpose of providing relevant reference basis for the research of the components of the Japanese thistle.)

1. The method for determining the chemical components of the cirsium japonicum charcoal based on the extraction and separation method is characterized by comprising the following steps of:

s1, preparing an extract: pulverizing 5kg of thistle charcoal, extracting with 70% ethanol under reflux for 3 times, mixing filtrates, and concentrating under reduced pressure to obtain 800g of extract;

s2, separating the extract; dissolving 800g of the extract in 8L of distilled water, sequentially extracting with petroleum ether, chloroform and ethyl acetate for 3 times respectively, and recovering the solvent to obtain 87g of petroleum ether part, 93g of chloroform part, 56g of ethyl acetate part and 90g of chloroform extraction part;

s3, elution and combination: separating the extract to obtain components, subjecting to silica gel column chromatography with chloroform and methanol gradient elution, and mixing into 8 parts, which are counted as Fr1-Fr 6;

s4, separation: performing Fr1 (5 g) column chromatography separation and purification to obtain compound 1 (780 mg), performing Fr2 (2 g) column chromatography separation and purification to obtain compound 3 (200 mg), performing Fr3 (6 g) column chromatography separation by silica gel column chromatography, performing cyclohexane-acetone gradient elution, performing acetone recrystallization purification to obtain compound 7 (110 mg), performing Fr4 (3.6 g) column chromatography separation and purification to obtain compound 5 (30 mg) and compound 8 (7 mg), dissolving 50g of ethyl acetate part in 500ml of distilled water, applying to D101 macroporous resin, performing gradient elution by ethanol with different contents, performing 30% ethanol elution part 18.9g column chromatography separation, applying ODS column methanol-water gradient elution to obtain compounds 2 (30 mg) and 4 (23 mg), and performing column chromatography separation by 70% ethanol part 3.9g to obtain compound 6 (45 mg).

2. The method for determining the chemical components of the cirsium japonicum charcoal based on the extraction and separation method as claimed in claim 1, wherein the compound 1 is yellow powder, the hydrochloric acid-magnesium powder reaction is positive, the Molish reaction is negative, and the compound is the pectolinarin.

3. The method for determining the chemical components of the cirsium japonicum charcoal based on the extraction and separation method as claimed in claim 1, wherein the compound 2 is yellow powder, the hydrochloric acid-magnesium powder reaction is positive, the Molish is positive, and the compound is the pectolinarin.

4. The method for determining the chemical components of the cirsium japonicum charcoal based on the extraction and separation method as claimed in claim 1, wherein the compound 3 is yellow powder, the hydrochloric acid-magnesium powder reaction is positive, the Molish reaction is negative, and the compound is acacetin.

5. The method for determining chemical components of Cirsium japonicum charcoal based on extraction and separation method as claimed in claim 1, wherein the compound 4 is pale yellow powder, and has positive reaction of hydrochloric acid-magnesium powder and positive reaction of Molish, and is robinin.

6. The method for determining the chemical components of the thistle charcoal based on the extraction and separation method as claimed in claim 1, wherein the compound 5 is yellow powder, the hydrochloric acid-magnesium powder reaction is positive, the Molish reaction is negative, and the thistle charcoal is diosmetin.

7. The method for determining the chemical components of Cirsium japonicum charcoal based on the extraction and separation method as claimed in claim 1, wherein the compound 6 is yellow powder, the hydrochloric acid-magnesium powder reaction is positive, the Molish reaction is negative, and the compound is quercetin.

8. The method for determining chemical components of Cirsium japonicum based on extraction and separation method as claimed in claim 1, wherein the compound 7 is yellow powder, the hydrochloric acid-magnesium powder reaction is positive, the Molish reaction is negative, and is trihydroxy-dimethoxy flavone.

9. The method for determining the chemical components of the cynum chinense Mill based on the extraction and separation method as claimed in claim 1, wherein the compound 8 is white needle crystal and catechol.

Technical Field

The invention relates to the technical field of thistle, in particular to a method for determining chemical components of thistle charcoal based on an extraction and separation method.

Background

The Japanese thistle herb is dry overground part or root of Japanese thistle herb of the Compositae, is cool in nature, sweet in taste and slightly bitter, enters heart and liver meridians, has the functions of cooling blood, stopping bleeding, removing blood stasis and eliminating carbuncle, is mainly used for treating various blood syndromes, leukorrhagia, female metrorrhagia and metrostaxis, sore and ulcer gall, scrofula, eczema, hepatitis, nephritis, furuncle and other diseases, is one of the main hemostatic medicaments in the clinical traditional Chinese medicine, and has long use history and definite curative effect. The Japanese thistle herb decoction pieces comprise Japanese thistle herb and Japanese thistle herb charcoal, and the main hemostatic effect is stir-frying charcoal at present. After stir-baked into charcoal, the properties, functions and appearance of Cirsium japonicum have changed greatly, and the change is necessarily related to the change of the intrinsic material basis.

At present, the compounds separated from the circium japonicum mainly comprise long-chain alkynol compounds, triterpenes, steroids, flavones, flavonoid glycosides and the like. The research on the chemical components of the cirsium japonicum charcoal is not reported, so that the production and the application of the cirsium japonicum are restricted to a certain extent.

An effective solution to the problems in the related art has not been proposed yet.

Disclosure of Invention

Aiming at the problems in the related art, the invention provides a method for determining the chemical components of the cirsium japonicum charcoal based on an extraction and separation method, so as to overcome the technical problems in the prior related art.

The technical scheme of the invention is realized as follows:

the method for determining the chemical components of the cirsium japonicum charcoal based on the extraction and separation method comprises the following steps:

s1, preparing an extract: pulverizing 5kg of thistle charcoal, extracting with 70% ethanol under reflux for 3 times, mixing filtrates, and concentrating under reduced pressure to obtain 800g of extract;

s2, separating the extract; dissolving 800g of the extract in 8L of distilled water, sequentially extracting with petroleum ether, chloroform and ethyl acetate for 3 times respectively, and recovering the solvent to obtain 87g of petroleum ether part, 93g of chloroform part, 56g of ethyl acetate part and 90g of chloroform extraction part;

s3, elution and combination: separating the extract to obtain components, subjecting to silica gel column chromatography with chloroform and methanol gradient elution, and mixing into 8 parts, which are counted as Fr1-Fr 6;

s4, separation: respectively separating and purifying Fr1 (5 g) column chromatography to obtain compound 1 (780 mg), separating Fr2 (2 g) column chromatography to obtain compound 3 (200 mg), separating Fr3 (6 g) by silica gel column chromatography, gradient eluting with cyclohexane-acetone, recrystallizing with acetone to obtain compound 7 (110 mg), separating Fr4 (3.6 g) column chromatography to obtain compound 5 (30 mg) and compound 8 (7 mg), dissolving ethyl acetate 50g in 500ml distilled water, loading onto D101 macroporous resin, gradient eluting with different content of ethanol, separating 30% ethanol eluate 18.9g column chromatography, loading onto ODS column methanol-water gradient eluting to obtain compound 2 (30 mg) and compound 4 (23 mg), separating 70% ethanol eluate 3.9g to obtain compound 6 (45 mg)

Further, the compound 1 is yellow powder, the hydrochloric acid-magnesium powder reaction is positive, the Molish reaction is negative, and the compound is pectolinarin.

Further, the compound 2 is yellow powder, the hydrochloric acid-magnesium powder reaction is positive, the Molish is positive, and the compound is pectolinarin.

Further, the compound 3 is yellow powder, the hydrochloric acid-magnesium powder reaction is positive, the Molish reaction is negative, and the compound is robinin.

Further, the compound 4 is light yellow powder, the hydrochloric acid-magnesium powder reaction is positive, the Molish reaction is positive, and the compound is robinin.

Further, the compound 5 is yellow powder, the hydrochloric acid-magnesium powder reaction is positive, the Molish reaction is negative, and the geraniol is obtained.

Further, the compound 6 is yellow powder, the hydrochloric acid-magnesium powder reaction is positive, the Molish reaction is negative, and the compound is quercetin.

Further, the compound 7 is yellow powder, the hydrochloric acid-magnesium powder reaction is positive, the Molish reaction is negative, and the compound is trihydroxy-dimethoxy flavone.

Further, compound 8 is white needle-like crystals and catechol.

The invention has the beneficial effects that: the method separates by means of macroporous adsorption resin and normal phase silica gel column chromatography, and determines the components of the compound by analyzing the data of the spectroscopy, thereby realizing the purpose of providing relevant reference basis for the research of the components of the Japanese thistle.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings without creative efforts.

FIG. 1 is a schematic flow chart of a method for determining chemical components of thistle carbon based on an extraction and separation method according to an embodiment of the invention.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments that can be derived by one of ordinary skill in the art from the embodiments given herein are intended to be within the scope of the present invention.

According to an embodiment of the present invention, a method for determining the chemical composition of Cirsium japonicum charcoal based on an extraction and separation method is provided.

As shown in figure 1, the method for determining the chemical components of the cirsium japonicum charcoal based on the extraction and separation method comprises the following steps:

the method comprises the following steps:

s1, preparing an extract: pulverizing 5kg of thistle charcoal, extracting with 70% ethanol under reflux for 3 times, mixing filtrates, and concentrating under reduced pressure to obtain 800g of extract;

s2, separating the extract; dissolving 800g of the extract in 8L of distilled water, sequentially extracting with petroleum ether, chloroform and ethyl acetate for 3 times respectively, and recovering the solvent to obtain 87g of petroleum ether part, 93g of chloroform part, 56g of ethyl acetate part and 90g of chloroform extraction part;

s3, elution and combination: separating the extract to obtain components, subjecting to silica gel column chromatography with chloroform and methanol gradient elution, and mixing into 8 parts, which are counted as Fr1-Fr 6;

s4, separation: performing Fr1 (5 g) column chromatography separation and purification to obtain compound 1 (780 mg), performing Fr2 (2 g) column chromatography separation and purification to obtain compound 3 (200 mg), performing Fr3 (6 g) column chromatography separation by silica gel column chromatography, performing cyclohexane-acetone gradient elution, performing acetone recrystallization purification to obtain compound 7 (110 mg), performing Fr4 (3.6 g) column chromatography separation and purification to obtain compound 5 (30 mg) and compound 8 (7 mg), dissolving 50g of ethyl acetate part in 500ml of distilled water, applying to D101 macroporous resin, performing gradient elution by ethanol with different contents, performing 30% ethanol elution part 18.9g column chromatography separation, applying ODS column methanol-water gradient elution to obtain compounds 2 (30 mg) and 4 (23 mg), and performing column chromatography separation by 70% ethanol part 3.9g to obtain compound 6 (45 mg).

Wherein, the compound 1 is yellow powder, the hydrochloric acid-magnesium powder reaction is positive, the Molish reaction is negative, and the compound is pectolinarin.

Wherein the compound 2 is yellow powder, the hydrochloric acid-magnesium powder reaction is positive, the Molish is positive, and the compound is pectolinarin.

Wherein, the compound 3 is yellow powder, the hydrochloric acid-magnesium powder reaction is positive, the Molish reaction is negative, and the compound is acacetin.

Wherein the compound 4 is light yellow powder, the hydrochloric acid-magnesium powder reaction is positive, the Molish reaction is positive, and the compound is robinin.

Wherein, the compound 5 is yellow powder, the hydrochloric acid-magnesium powder reaction is positive, the Molish reaction is negative, and the geraniin is obtained.

Wherein, the compound 6 is yellow powder, the hydrochloric acid-magnesium powder reaction is positive, the Molish reaction is negative, and the compound is quercetin.

Wherein, the compound 7 is yellow powder, the hydrochloric acid-magnesium powder reaction is positive, the Molish reaction is negative, and the compound is trihydroxy-dimethoxy flavone.

Wherein the compound 8 is white needle crystal and is catechol.

In conclusion, according to the technical scheme of the invention, the components of the compound are determined by separating through macroporous adsorption resin and normal phase silica gel column chromatography and analyzing the data of the spectroscopy, so that the related reference basis is provided for the research of the components of the circium japonicum.

The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.