A method for separating and purifying ginsenotriol

Abstract

The present invention relates to a method for purifying ginsenoside triol, after danshen is dried, sliced, then ground into 50-100 mesh powder, danshen powder is extracted by ethanol heating reflux, then the extract is dried by rotary evaporation, after the solid obtained is extracted with ethyl acetate 1-2 times, the extract is dried by rotary evaporation to obtain a crude product of ginsenoside triol. After ion exchange resin is treated with calcium chloride aqueous solution, clean water is washed and dried, then a column is loaded to obtain a resin column, after the crude product of ginsenoside triol is dissolved with ethyl acetate, the resin column is loaded, first rinsed with ethyl acetate, then rinsed with an ethanol aqueous solution of calcium chloride, the rinse solution is collected, and the rotary evaporation is dried to obtain. The method utilizes the characteristics of vicinal diols in ginsenoside triol combined with metal ions to carry out affinity purification, compared with existing methods, simple to operate, short time consumption, the purification efficiency and yield of ginsenoside triol are relatively high, and it is suitable for industrial production.

Description

Technical Field

[0001] The invention relates to a method for separating and purifying ginsenotriol, and belongs to the technical field of medicines. Background Art

[0002] Due to environmental pollution and aging, the incidence and mortality of cancer in my country are increasing year by year. According to the current medical level, once diagnosed with cancer, most patients will receive chemotherapy or surgery plus chemotherapy. However, in addition to killing cancer cells, chemotherapy drugs also cause serious damage to the normal body. Clinically, the side effects of anticancer drugs are manifested in decreased white blood cells, diarrhea, and often accompanied by rapid weight loss, which seriously affect the quality of life of patients during chemotherapy. At present, there are not many auxiliary drugs on the market to improve the side effects of chemotherapy and improve the quality of life of patients during chemotherapy. Only interleukins are used to increase white blood cells to compensate for the decline in immunity during chemotherapy. Patients still have problems with decreased appetite and weight loss.

[0003] In recent years, American researchers have found that ginsenoside can effectively protect body tissues and prevent animals from losing weight when used with chemotherapy drugs. These findings have also been published in influential journals, such as the Proceedings of the National Academy of Sciences (PNAS). These research results show that it is very feasible to use ginsenoside as an adjuvant drug for chemotherapy or as a daily health product for chemotherapy patients to improve the quality of life of chemotherapy patients. However, the content of ginsenoside is low in various types of ginseng, and its structure determines that it is not easy to separate and purify, which affects its industrial production and application.

[0004] At present, the separation and purification method of panaxylacettriol is usually to extract with ethyl acetate at room temperature for 20-30h, then use ethyl acetate-n-hexane (1:1, V / V) as eluent, separate the ethyl acetate extract by column chromatography, and then use chloroform and ethyl acetate-n-hexane for further separation, but this method is time-consuming, low in yield, and uses chloroform that is seriously toxic to the human body. In the prior art, there is also a method of using a silica gel column to separate and purify the ethyl acetate extract, washing the column with ethyl acetate and n-hexane, collecting the washing liquid, and preparing panaxylacettriol, but this method has the problem of low yield and purification efficiency. Summary of the invention

[0005] The purpose of the present invention is to solve the deficiencies of the prior art and provide a method for separating and purifying ginsenotriol. The method is simple to operate, has high purification efficiency and yield of ginsenotriol, and is suitable for industrial production.

[0006] Technical Solution

[0007] A method for separating and purifying ginsenotriol comprises the following steps:

[0008] (1) drying and slicing Danshen, and then grinding into powder of 50-100 mesh to obtain Danshen powder;

[0009] (2) extracting the Danshen powder by heating and refluxing with ethanol, and then drying the extract by rotary evaporation, extracting the obtained solid with ethyl acetate for 1-2 times, and then drying the extract by rotary evaporation to obtain a crude product of ginsenoside triol;

[0010] (3) treating the ion exchange resin with a calcium chloride aqueous solution, washing with clean water and drying, and then loading the resin into a column to obtain a resin column;

[0011] (4) After dissolving the crude ginsenoside triol obtained in step (2) in ethyl acetate, the crude ginsenoside triol is loaded onto the resin column in step (3), firstly rinsed with ethyl acetate and then rinsed with an ethanol aqueous solution of calcium chloride, the rinse liquid is collected, and rotary evaporated to dry to obtain ginsenoside triol.

[0012] Furthermore, in step (2), the extraction by heating and refluxing with ethanol is performed twice, and each extraction lasts for 1-2 hours.

[0013] Furthermore, in step (3), the method of treating with calcium chloride aqueous solution is: adding calcium chloride aqueous solution to ion exchange resin, soaking overnight, filtering, adding calcium chloride aqueous solution to ion exchange resin again, soaking overnight, and filtering; the calcium chloride aqueous solution is prepared by dissolving 50g of calcium chloride in 100g of water.

[0014] Furthermore, in step (3), the drying is carried out in a drying oven at 50% humidity, at a temperature of 25-35° C., and for 5-7 days.

[0015] Furthermore, in step (4), the ethanol aqueous solution of calcium chloride is a saturated calcium chloride solution prepared by using a mixture of ethanol and water in a volume ratio of 7:3 as a solvent.

[0016] Beneficial effects of the present invention:

[0017] The invention provides a method for separating and purifying ginsenoside triol, which utilizes the characteristics of vicinal diol in ginsenoside triol binding to metal ions for affinity purification. Compared with the existing method, the method is simple to operate, short in time, and has relatively high purification efficiency and yield of ginsenoside triol, and is suitable for industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 The NMR spectrum of panaxytriol extracted in Example 1;

[0019] Figure 2 This is the MS spectrum of panaxytriol extracted in Example 1. DETAILED DESCRIPTION

[0020] The technical solution of the present invention is further described below in conjunction with the accompanying drawings and specific embodiments.

[0021] Example 1

[0022] A method for separating and purifying ginsenotriol comprises the following steps:

[0023] (1) drying and slicing Danshen, and then grinding into powder of 50-100 mesh to obtain Danshen powder;

[0024] (2) 350 g of Danshen powder and 700 mL of ethanol were mixed, heated under reflux for extraction for 1 h, the extract was separated, and then heated under reflux for extraction with 700 mL of ethanol for 1 h. The extracts were combined and dried by rotary evaporation. The obtained solid was extracted with 400 mL and 200 mL of ethyl acetate, respectively, the extracts were combined, and then dried by rotary evaporation to obtain 3.65 g of crude ginsenoside triol;

[0025] (3) adding a calcium chloride aqueous solution to the ion exchange resin, soaking it overnight, filtering it, adding a calcium chloride aqueous solution (prepared by dissolving 50 g of calcium chloride in 100 g of water) to the ion exchange resin again, soaking it overnight, filtering it, washing it with clean water and drying it, the drying is carried out in a drying oven with a humidity of 50%, drying it at 28° C. for 7 days, and then loading it into a column (column diameter 2.5 cm, length 3.5 cm), to obtain a resin column;

[0026] (4) 3.65 g of the crude product of ginsenotriol obtained in step (2) was dissolved in ethyl acetate and loaded onto the resin column of step (3), first rinsed with ethyl acetate and then rinsed with an ethanol aqueous solution of calcium chloride (a saturated solution of calcium chloride prepared with a mixture of ethanol and water in a volume ratio of 7:3 as a solvent at 20° C.), the rinse liquid was collected, and rotary evaporated to dry to obtain 800 mg of ginsenotriol.

[0027] Figure 1 is the NMR spectrum of panaxytriol extracted in Example 1, Figure 2 This is the MS spectrum of ginsenotriol extracted in Example 1. It can be seen that the component of the extract is ginsenotriol with high purity (>85%).

Claims

1. A method for separating and purifying ginsenotriol, characterized in that: The steps include: (1) drying and slicing Danshen, and then grinding into powder of 50-100 mesh to obtain Danshen powder; (2) extracting the Danshen powder by heating and refluxing with ethanol, and then drying the extract by rotary evaporation, extracting the obtained solid with ethyl acetate for 1-2 times, and then drying the extract by rotary evaporation to obtain a crude product of ginsenoside triol; (3) treating the ion exchange resin with a calcium chloride aqueous solution, washing with clean water and drying, and then loading the resin into a column to obtain a resin column; (4) After dissolving the crude ginsenoside triol obtained in step (2) in ethyl acetate, the crude ginsenoside triol is loaded onto the resin column in step (3), firstly rinsed with ethyl acetate and then rinsed with an ethanol aqueous solution of calcium chloride, the rinse liquid is collected, and rotary evaporated to dry to obtain ginsenoside triol.

2. The method for separating and purifying ginsenotriol according to claim 1, characterized in that: In step (2), the extraction is performed twice by heating with ethanol under reflux, and each extraction lasts for 1-2 hours.

3. The method for separating and purifying ginsenotriol according to claim 1, characterized in that: In step (3), the method of treating with calcium chloride aqueous solution is: adding calcium chloride aqueous solution to ion exchange resin, soaking overnight, filtering, adding calcium chloride aqueous solution to ion exchange resin again, soaking overnight, filtering; the calcium chloride aqueous solution is prepared by dissolving 50g of calcium chloride in 100g of water.

4. The method for separating and purifying ginsenotriol according to claim 1, characterized in that: In step (3), the drying is carried out in a drying oven at 50% humidity, at a temperature of 25-35° C., for 5-7 days.

5. The method for separating and purifying ginsenotriol according to claim 1, 2, 3 or 4, characterized in that: In step (4), the ethanol aqueous solution of calcium chloride is a saturated calcium chloride solution prepared by using a mixture of ethanol and water in a volume ratio of 7:3 as a solvent.

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