WO2004082695A1

WO2004082695A1 - Shenfu sterile powder and the preparation thereof

Info

Publication number: WO2004082695A1
Application number: PCT/CN2003/000190
Authority: WO
Inventors: Hai Yin; Yanming Zeng; Jilin Sun
Original assignee: Sichuan Dikang Industry Holdings Group Co., Ltd.
Priority date: 2003-03-17
Filing date: 2003-03-17
Publication date: 2004-09-30
Also published as: AU2003221295A1; CN1720056A

Abstract

The present invention discloses a Shenfu sterile powder, which comprises the active component extract of radix red ginseng and the active component extract of radix aconiti praeparata, characterized in that the two extracts were isolated by water extraction, then removing impurities by absorbed resin column, filtering through refine decolor resin column, and drying to obtain Shenfu sterile powder. Both the content of ginsenosides and the content of alkaloids from radix aconiti praeparata are more than 50%.

Description

Reference attached sterile powder and preparation method thereof

The invention belongs to the field of pharmacy, and relates to a ginseng-containing sterile powder including a Chinese medicinal material red ginseng and aconite extracting effective parts, and a preparation method thereof. Background technique

The existing ginsenosides in active parts of red ginseng are generally extracted by refluxing with ethanol. The disadvantages of this method of extraction are: the color of the extraction is deep, the impurities, especially the fat-soluble impurities are many, the purity of the pure dagger is difficult, the process is relatively complicated, and the cost is high.

The existing aconite alkaloids in effective parts of aconite are usually extracted by refluxing with ethanol. This method extracts aconite alkaloids with many pigment impurities, especially fat-soluble impurities. In addition, the effective content of aconite active ingredients is low, and subsequent purification is very difficult. The operation is complicated and costly.

As a result, the prepared Shenfu freeze-dried powder injection has a lot of impurities, which directly affects the efficacy of the medicine and the temperature of the product. Disclosure of invention

The purpose of the present invention is to provide a ginseng aseptic powder containing red ginseng and aconite extracting effective parts with a scientific preparation process, scientific mixing ratio, controllable quality, and low extraction cost.

Another object of the present invention is to provide a method for preparing aseptic powder.

In order to achieve the above object, the technical solution of the present invention is: a ginseng-attached sterile powder, which includes a red ginseng effective part extract and an aconite active part extract, characterized in that the red ginseng effective part extract is obtained by the following method: Weigh the required amount of red ginseng, add 10-20 times water to boil and keep at 80-90 ° C for 2 hours, filter, and drug residue add 10 to 20 times water to boil at 80-90 ° C and keep 1. After 5 hours, filter, boil the residue with 10-20 times water and boil at 80-90 ° C for 1 hour, filter, combine the filtrates, and remove the precipitate by speed centrifugation to obtain the ginseng water extract; the ginseng water extract passes through Adsorption resin chromatography column. After adsorption, wash with 0.1-10% alkaline solution until colorless, and then wash with distilled water until the pH is neutral, elute with 75-85% ethanol, until no saponin reaction, elute The subsequent ethanol solution was passed through a purified decolorized resin chromatography column. The effluent was collected and left to stand for 12-24 hours, filtered, and concentrated under reduced pressure to ethanol-free, filtered to obtain a transparent light yellow red ginseng extract, frozen under aseptic conditions. Dry to obtain a dry extract of the effective part of red ginseng, and the total saponin content of ginseng>50%; the extract of the effective part of aconite is obtained by the following method: Weigh the required amount of aconite, add 10-20 times PH3-9 water and boil it at 80-90 ° C Hold for 2 hours, filter, boil water with 10-20 times PH3-9 and boil at 80-90 ° C for 1.5 hours, filter, boil water with 10-20 times PH3-9 Hold at 80-90 ° C for 1 hour, filter, combine the filtrates, and remove the precipitate by speed centrifugation to obtain the aconite aqueous extract. The aconite aqueous extract is passed through the adsorption resin chromatography column and washed with water until the eluate is colorless. -85% ethanol elutes to no alkaloid reaction, and passes through the purified decolorized resin chromatography column at a uniform speed. The eluate is left to be filtered and concentrated under reduced pressure to ethanol-free; filtered and freeze-dried under sterile conditions to obtain solids in the effective part of aconite. Extract containing aconite alkaloids> 50%.

The ginseng-containing sterile powder of the present invention comprises 10-20 parts by weight of the active part extract of red ginseng; 0.1-3.0 parts by weight of the active part extract of aconite.

The invention relates to a method for preparing a reference-attached sterile powder, which comprises the following steps:

1) Extract the extract of the effective part of red ginseng: Weigh the required red ginseng, boil it with 10-20 times water and boil at 80-90 ° C for 2 hours, filter, and boil the medicine residue with 10-20 times water Hold at 80-9 ° C for 5 hours, filter, boil the residue with 10-20 times water and boil at 80-90 ° C for 1 hour, filter, combine the filtrates, and remove the precipitate by speed centrifugation to obtain ginseng water extract. Fluid

2) Extract of effective part of aconite: Weigh the required aconite, add 10-20 times PH3-9 water, boil it and keep it at 80-90 ° C for 2 hours, filter, add medicine residue 10-20 times PH3-9 After boiling, keep at 80-90 ° C for 1.5 hours, and filter. The residue is boiled with water at 10-20 times PH3-9. After boiling, keep at 80-90 ° C for 1 hour, filter, and combine the filtrates. Speed flow centrifugation Remove the sediment to obtain aconite water extract;

3) Purification of ginseng water extract: The ginseng water extract passed through the adsorption resin chromatography column. After adsorption, it was washed with 0.1-10% alkaline solution to colorless, and then washed with distilled water until the pH was neutral. Elute with 75-85% ethanol until saponin-free reaction. The eluted ethanol solution is passed through a purified decolorizing resin chromatography column uniformly. The effluent is collected and left to stand for 12-24 hours, filtered, and the reduced pressure film is concentrated to ethanol-free, filtered. , Obtaining a transparent light yellow red ginseng extract, freeze-dried under aseptic conditions to obtain a dry extract of the effective part of red ginseng, and the total ginsenoside content is> 50%;

4) Purification of aconite water extract: The aconite water extract is passed through the adsorption resin chromatography column uniformly, washed with water until the eluent is colorless, and eluted with 75-85% ethanol until there is no alkaloid reaction, and the decolorized resin layer is purified at a uniform speed. Analytical column, the eluate is left to be filtered, and concentrated under reduced pressure to ethanol-free; filtered, freeze-dried under aseptic conditions to obtain a solid extract of the effective part of aconite, containing aconite alkaloid> 50%. 5) Weigh 10-20 parts by weight of the extract from the active part of red ginseng; 0.1-3.0 parts by weight of the extract from the active part of aconite.

Experiment 1: Effect of extraction method on ginsenoside extraction rate

Existing ethanol to extract red ginseng in parallel: Red ginseng is dried in an oven at 80 ° C for 12 hours, crushed with a pulverizer, passed through a 20-mesh sieve, and dipped in ethanol in accordance with the method of ginseng saponin injection of Sichuan Pharmaceutical Standard To extract red ginseng, specifically weigh 100g of red ginseng powder and put it in a flask, soak it in 300ml of ethanol for 72 hours, filter, and use the filtrate for backup. Add 300ml of ethanol residue to the water bath and reflux for three times, 2 hours each time. Measure ginsenoside content.

Extraction of red ginseng using the water of the present invention: Weigh 100g of red ginseng powder into a flask, add 100ml of distilled water, and boil it at 80-90 ° C for 2 hours, filter, and use the filtrate for later use. Extract the drug residue with 100ml of distilled water twice according to the aforementioned method, and the extraction time is 1.5 and 1 hour, respectively. The filtrate was sampled separately to determine the ginsenoside content.

Each treatment was repeated 3 times. The experimental results are shown in Table 1. It shows that there is no significant difference in the content of ginsenosides extracted from the water-boiled ginsenosides and ethanol from the present invention, and the water-boiled extraction method is only slightly lower than the ethanol extraction. However, the ethanol extraction process is complicated and the cost is much higher. Compared with the water extraction method, it is better to use the water boiling method.

Table 1 Effect of extraction method on ginsenoside extraction rate Extraction method Each extraction rate Total

1 2 3 (g / noodle g) 51.6% 26% 18% 95.6% 1.75

Ethanol extraction 58% 27% 15% 100% 1.83 Experiment 2: Effect of extraction methods on extraction rate of aconite alkaloids

Extraction of aconite from ethanol under reflux: Aconite is dried in an oven at 60 ° C for 12 hours, crushed with a pulverizer, passed through a 20-mesh sieve, weighed 100g of aconite powder and placed in a clean steaming bottle, and added 80% ethanol to 500ml. Hours, filter for backup, and add 250% 80% ethanol to 250 ml reflux extraction for each time, each time is 1.5 and 1 hour, after each reflux sampling, extract extract aconite alkaloid content.

Extraction of aconite alkaloids using the water of the present invention: Weigh 100g of aconite powder into a steamer bottle, add 1L of distilled water, and boil it for 2 hours at 80-9 ° C, filter, and use the filtrate as a reserve. Drug residue plus distilled water 1L was leached twice according to the aforementioned method, and the leaching times were 1. 5 and 1 hour. Samples were taken for alkaloid content after each boiling step.

Each treatment was repeated 3 times, and the experimental results are shown in Table 2. It is shown that the content of aconite alkaloids extracted by water-boiled aconite in the present invention is not significantly different from the content of aconite alkaloids extracted with ethanol under reflux. The water-boiled extraction method is only slightly lower than the ethanol extraction. However, the ethanol extraction process is complicated and the cost is much higher. Compared with water extraction, the extraction method is better.

Table 2 Effect of extraction methods on extraction rate of aconite alkaloids Extraction methods Each extraction rate Total alkaloid ratio

1 2 3 (/ 100g) Boiled extraction 54% 26% 17.6% 97.6% 0.189 Ethanol reflux 67% 25% 8% 100% 0.228 Experiment 3 Purification of Ginseng Aconite Extract

Decoloring resin column control parameters: 25ml alcohol solution / g decolorizing resin. After the decolorizing resin, the red ginseng and aconite extracts were 29.5L and 28.8L, respectively, and they were left overnight for 12 hours. Filter through a No. 4 glass sand funnel, and then filter with a No. 5 glass sand funnel. The membrane is concentrated under reduced pressure (temperature §0-90 ° C, vacuum degree 68cm), and alcohol-free. Under aseptic conditions, it was filtered with a No. 5 glass sand funnel and freeze-dried to obtain 180g of red ginseng total saponins and 18.8g of aconite total alkaloids, respectively, and leaching with alcohol under reflux and purification in the above manner to obtain red ginsenosides and Aconite alkaloids are 16¾ and 15J5g, respectively. It is slightly lower than the water extraction method, which is mainly due to the large amount of impurities and the large loss of the effective part during purification. The results of three batches of purification and extraction are shown in Tables 3 and 4.

Table 3 Purification and extraction results of three batches of red ginseng total saponins

Three batches of ginsenosides were checked for hemolysis, heat source, microorganisms, clarity, etc. according to the requirements of the National Pharmacopoeia "Injection", and the results showed that three batches of total ginsenosides were at a concentration of 5mg / ml, and no hemolysis occurred after 4 hours The heat source, clarity, and microbiological inspections are in full compliance with the requirements. Table 4 Purification results of three batches of aconite alkaloid extract

Three batches of aconite were inspected for hemolysis, heat source, microorganisms, clarity, etc. according to the requirements of the National Pharmacopoeia "Injection". The results showed that the total alkaloids of three batches of aconite were 2.5mg / ml, and no hemolysis was observed for 4 hours. Occurrence, heat source, clarity, and microbiological inspections are fully satisfactory. Experiment 4 Quality inspection of three batches of injection-attached sterile powder

Three batches of 1000 ginseng-containing sterile powders for trial production were produced. Three batches of samples were sampled at a sampling rate of 1%, and inspected in accordance with the requirements of the National Pharmacopoeia "Injectables", and the results showed that the requirements of the National Pharmacopoeia were met. See table 5

* Resin content is measured by resin-color method

^ Developing agent: n-butanol-ethyl acetate-water (4 .. The best way to implement the present invention

Example 1

1) Take the extract of the effective part of red ginseng: Weigh the required red ginseng, add 20 times water and boil it at 80. C is kept for 2 hours, filtered, boiled with 20 times water and boiled at 80-° C for 1.5 hours, filtered, boiled with 20 times water and boiled at 80. C was kept for 1 hour, filtered, the filtrates were combined, and the precipitate was removed by speed centrifugation to obtain a water extract of ginseng; 2) extract of effective part of aconite: weigh the required aconite, add 20 times PH3 water and boil it at 80 ° C and keep 2 hours, filtered, boiled with 20 times PH9 water and boiled at 90 ° C for 1.5 hours, filtered, boiled with boiled water and 10 times PH3 water

Keep it at 80 ° C for 1 hour, filter, combine the filtrates, remove the precipitate by speed centrifugation to obtain aconite extract; 3) Purification of ginseng water extract: The ginseng water extract passes through the adsorption resin chromatography column. After the adsorption is completed, it is washed with 0.1% alkaline solution until it is colorless, and then washed with distilled water to neutralize pH. Elute with 75% ethanol until there is no saponin reaction. After the elution, the ethanol solution is passed through a purified decolorizing resin chromatography column uniformly. The effluent is collected and allowed to stand for 12 hours, filtered, and the reduced pressure film is concentrated to ethanol-free. Yellow red ginseng extract, frozen and dried under aseptic conditions to obtain a dry extract of the effective part of red ginseng, the total saponin content of ginseng>50%;

4) Purification of aconite water extract: The aconite water extract is passed through the adsorption resin chromatography column uniformly, washed with water until the eluent is colorless, and eluted with 85% ethanol until no alkaloid reaction, and uniformly passed through the purified decolorized resin chromatography column. The eluate was left to be filtered, and concentrated under reduced pressure to ethanol-free; filtered, and freeze-dried under a sterile glass to obtain a solid extract of the effective part of aconite, containing aconite alkaloids> 50%.

5) Weigh 100g of auxiliary materials, add 450ml of water for injection, stir to dissolve, and then weigh 10g of active part extract of red ginseng, 3.0g of active part extract of aconite, add water to make up to 1000ml, gently stir until all dissolved, Under pressure, filter the medicinal solution with two filters in series and freeze-dry under aseptic conditions to make sterile powders for reference and aliquot them.

Example 2

1) Extract of active part of red ginseng: Weigh the required red ginseng, add 10 times of water to boil and hold it at 90 ° C for 2 hours, filter, and add 10 times of water to boil at 90 ° C. 1. After 5 hours of filtration, the residue was boiled with 10 times more water and boiled at 9 ° C for 1 hour, filtered, the filtrates were combined, and the precipitate was removed by speed centrifugation to obtain a ginseng water extract;

2) Extract of effective part of aconite: Weigh the required aconite, add 10 times of PH9 water to boil, and keep it at 90 ° C for 2 hours, filter, and add 10 times of PH3 water to boil at 90 ° C. After 5 hours of filtration, the residue was boiled with 10 times PH3 of water and boiled at 90 ° C for 1 hour. After filtration, the filtrates were combined and the precipitate was removed by speed centrifugation to obtain an aqueous extract of aconite;

3) Purification of ginseng water extract: The ginseng water extract passes through the adsorption resin chromatography column. After the adsorption is completed, it is washed with 10% alkaline solution until it is colorless, and then washed with distilled water until the pH is neutral, and 85% ethanol is used. Elution until saponin-free reaction. After the elution, the ethanol solution is passed through a purified decolorized resin chromatography column uniformly. The effluent is collected and allowed to stand for 24 hours, filtered, and concentrated under reduced pressure to ethanol-free. Filtered to obtain a transparent light yellow red ginseng extract. Liquid, freeze-dried under aseptic conditions to obtain a dry extract of the effective part of red ginseng, the total saponin content of ginseng> 50%;

4) Purification of aconite water extract: The aconite water extract is passed through an adsorption resin chromatography column with water, Wash until the eluent is colorless, and elute with 75% ethanol until there is no alkaloid reaction. Pass the purified decolorized resin chromatography column at a uniform speed. The eluent is left to stand for filtration, and concentrated under reduced pressure to ethanol-free; filtered, frozen under sterile conditions. Dry to obtain a solid extract of the effective part of aconite, containing aconite alkaloids> 50%.

5) Weigh 200g of auxiliary materials, add 500ml of water for injection, stir to dissolve, and then weigh 20g of active part extract of red ginseng, O.lg of effective part extract of aconite, add water to make up to 1000ml, gently stir until all dissolved, Under the condition of positive pressure, the medicinal solution is filtered with two filters connected in series, and freeze-dried under aseptic conditions to make sterile powders attached to the package and divided into packages.

Claims

Rights request

A ginseng-containing sterile powder, which includes an active part extract of red ginseng and an effective part extract of aconite, is characterized in that the effective part extract of red ginseng is obtained by the following method: Weigh the required amount of red ginseng, add 10 -After boiling for 20 times, keep it at 80-90 ° C for 2 hours, filter it, and add 10-20 times for boiling, keep it at 80-90 ° C for 1.5 hours, filter, add residue-10- After boiling 20 times, keep it at 80-90 ° C for 1 hour, filter, combine the filtrates, and remove the precipitate by speed centrifugation to obtain the ginseng water extract. The ginseng water extract passes through the adsorption resin chromatography column. After the adsorption, Wash with 0.1-10% alkaline solution until it is colorless, then wash with distilled water to neutral pH, elute with 75-85% ethanol, until no saponin reaction, the eluted ethanol solution is uniformly purified over the decolorization resin Chromatographic column, collect the effluent and let stand for 12-24 hours, filter, concentrate under reduced pressure to ethanol-free, filter, obtain transparent light yellow red ginseng extract, freeze-dry under sterile conditions, and obtain dry extract of effective part of red ginseng , Total ginsenosides content> 50%; aconite effective Bit extract is obtained by the following method: taking a desired amount of said tuber, plus 10--20 fold PH3 - 9 after boiling in water 80-90. C is kept for 2 hours, filtered, boiled with 10-20 times PH3-9 water and boiled at 80-90 ° C for 1.5 hours, filtered, boiled with 10-20 times PH3-9 water and boiled Hold at 80-90 ° C for 1 hour, filter, combine the filtrates, and remove the precipitate by speed centrifugation to obtain aconite aqueous extract. The aconite aqueous extract is passed through the adsorption resin chromatography column uniformly, and washed with water until the eluate is colorless. -85% ethanol elutes to no alkaloid reaction, and passes through the column of purified decolorized resin at a uniform speed. The eluate is left to be filtered, and concentrated under reduced pressure to ethanol-free. Filtration and freeze-drying under sterile conditions to obtain solids in the effective part of aconite Extract containing aconite alkaloids> 50%.

The aseptic powder according to claim 1, characterized in that: it comprises 10 to 20 parts by weight of the extract of the active part of red ginseng; 0.1-3.1 parts by weight of the extract of the effective part of aconite.

A method for preparing a sterile powder, including the following steps:

1) Take the extract of the effective part of red ginseng: Weigh the required red ginseng, boil it with 10-20 times water and boil it at 80-90 ° C for 2 hours, filter, and boil the medicine residue with 10-20 times water Hold at 80-90 ° C for 1.5 hours, filter, and add 10-20 times water to boil the residue at 80-

Hold at 90 ° C for 1 hour, filter, combine the filtrates, and remove the sediment by speed centrifugation to obtain a ginseng water extract;

2) Extract of effective part of aconite: Weigh the required aconite, add 10-20 times PH3-9 water, boil it and keep it at 80-90 ° C for 2 hours, filter, add medicine residue 10-20 times PH3-9 After boiling, keep at 80-90 ° C for 1.5 hours, and filter. The medicine residue is boiled with water at 10-20 times PH3-9. After boiling, keep at 80-90 ° C for 1 hour, filter, and combine the filtrates. Speed flow centrifugation Remove the sediment to obtain aconite water extract;

3) Purification of ginseng water extract: The ginseng water extract passed through the adsorption resin chromatography column at a uniform speed, and the adsorption was completed. After that, it was washed with 0.1-10% alkaline solution until it was colorless, and then washed with steamed maid water until the pH was neutral, and then eluted with 75-85% ethanol until no saponin was reacted. The eluted ethanol solution was uniformly purified over time. Decolorized resin chromatography column, collect the effluent and let stand for 12-24 hours, filter, and concentrate under reduced pressure to ethanol-free; filter to obtain transparent light yellow red ginseng extract, freeze dry under aseptic conditions, and obtain the effective part of red ginseng Extract, total ginsenoside content>50%;

) Purification of aconite water extract: The aconite water extract was passed through the adsorption resin chromatography column uniformly, washed with water until the eluent was colorless, and eluted with 75-85% ethanol until there was no alkaloid reaction. The column and the eluate were left to stand for filtration, and concentrated under reduced pressure to ethanol-free; filtered, and freeze-dried under aseptic conditions to obtain a solid extract of the effective part of aconite, containing aconite alkaloids> 50%.

) Weigh 10 to 20 parts by weight of the extract from the active part of red ginseng; 0.1-3.0 parts by weight of the extract from the effective part of aconite.