CN111825779A - Extraction method of Dendrobium officinale polysaccharide - Google Patents

Abstract

The invention provides an extraction method of dendrobium officinale polysaccharide, which is a novel extraction method of dendrobium officinale polysaccharide, and specifically comprises the following steps: pulverizing dried herba Dendrobii, weighing appropriate amount, adding anhydrous ethanol, and sealing and soaking; after removing the ethanol, adding distilled water according to a certain feed-liquid ratio, and setting ultrasonic time, extraction temperature, extraction time, ultrasonic power and extraction times for extraction; after sufficient extraction, the polysaccharide solution is concentrated under reduced pressure, and the impurities are removed by using absolute ethyl alcohol. The results show that: the yield of the dendrobium officinale polysaccharide is 61.12%. The invention solves the comprehensive problem that the extraction rate and production are influenced because the optimal process condition cannot be determined due to the complex extraction method and excessive influence factors during the extraction of the dendrobium officinale polysaccharide.

Description

Extraction method of Dendrobium officinale polysaccharide

technical field

The invention belongs to the field of extraction of effective components in plants, in particular relates to various factors affecting the extraction rate of Dendrobium officinale, and realizes a method for extracting Dendrobium officinale polysaccharide with optimal experimental parameters determined under comprehensive influencing factors. The Dendrobium officinale polysaccharide can obtain a high extraction rate under a simple extraction process, and has stable work, low energy consumption, and can be mass-produced.

Background technique

Dendrobium officinale (Dendrobium officinale) is an important medicinal and edible plant, which can benefit the stomach and promote fluid, immune regulation and delay aging. Studies have shown that Dendrobium officinale contains a variety of active ingredients such as polysaccharides, alkaloids, polyphenols, flavonoids and amino acids, among which Dendrobium officinale polysaccharides (DOP) is the main active ingredient and the substance with the highest content, which has antibacterial, Enhance immunity, lower blood sugar, blood lipids and blood pressure, anti-tumor and antioxidant effects. A large number of studies have reported that the extraction methods currently used for DOP mainly include hot water extraction, enzymatic extraction, ultrasonic-assisted extraction, flash extraction, ultra-high pressure extraction and microwave-assisted extraction. Due to the complexity of the extraction process and the low extraction rate, selecting a simple extraction process while ensuring the extraction rate has become a research hotspot today. Hot water extraction is the most commonly used method for extracting Dendrobium officinale polysaccharides, but the extraction rate is not high. The intervention of ultrasonic waves will greatly improve the extraction efficiency. However, the magnitude of ultrasonic power has not been explored. Therefore, the determination of a polysaccharide extraction process of Dendrobium officinale determined by the best experimental parameters under the comprehensive influencing factors has a high development prospect.

SUMMARY OF THE INVENTION

The technical problem to be solved in the present invention is to provide an optimal extraction process of Dendrobium officinale polysaccharide, so as to overcome the above-mentioned defects in the prior art.

The present invention solves its technical problem and adopts following technical scheme:

The invention provides a method for extracting Dendrobium officinale polysaccharide, specifically: adding anhydrous ethanol to the dried and crushed Dendrobium officinale, sealing and soaking; adding distilled water after removing the ethanol, and then performing ultrasonic extraction and extraction to depressurize the obtained polysaccharide solution Concentrate, then use absolute ethanol for impurity removal.

The particle size of the Dendrobium officinale is 0.1-0.2 mm, 4.8-5.2 g of Dendrobium officinale is weighed, 48-52 mL of absolute ethanol is added, then sealed with a plastic wrap, and soaked for 11-13 hours.

The method for extracting Dendrobium officinale polysaccharide provided by the invention may include the following steps:

Step 1. Remove the absolute ethanol in the soaked sample, and add the sample and distilled water into a three-necked flask in an appropriate ratio;

Step 2. Set the working parameters of the ultrasonic cleaner, stir and extract;

Step 3. Filter the extracted solution into a conical flask, place the filtrate in an ultrasonic cleaner to sonicate, and use an electric heating temperature control device for heating. to centrifuge;

Step 4. Take the supernatant, add absolute ethanol for alcohol precipitation, let stand overnight, centrifuge to get the precipitate, coarsely filter the extracted solution into a 1000mL conical flask with a 100-mesh gauze, add pure water to rinse several times to dissolve Precipitation (filter cake);

Step 5. Add anhydrous ethanol to the above solution for secondary alcohol precipitation, let it stand in the refrigerator, centrifuge to get the precipitate, and add pure water to dissolve;

Step 6. After fully dissolving, centrifuge the solution and put the supernatant in a beaker;

Step 7. The polysaccharide solution is freeze-dried to obtain the target Dendrobium officinale polysaccharide.

In step 1 of the above method, the sample is mixed with distilled water at 1:20--1:60 (g/mL), preferably 1:50 (g/mL), and added to a 500mL three-necked flask.

In step 2 of the above method, a 240W ultrasonic cleaner is used, and its parameters are set as: heating temperature 50-90 ° C, heating time 120-240 min, ultrasonic time 30-150 min and ultrasonic power 20-100%, to perform stirring extraction.

In step 2 of the above method, a 240W ultrasonic cleaner is used, and its parameters are preferably set as follows: heating temperature 80°C, heating time 210min, ultrasonic time 120min and ultrasonic power 60% (140W) for stirring and extraction.

In step 3 of the above method, the extracted solution can be roughly filtered into a 1000 mL conical flask with a 100-mesh gauze, and the filter cake is washed with pure water several times, generally 3 times.

In step 3 of the above method, the concentrated solution can be centrifuged at 3000-4500r/min for 10-20min. Preferably, the concentrate is centrifuged at 4500r/min for 10min.

In step 4 of the above method, add anhydrous ethanol to the supernatant to 300-500mL, let stand in a 4°C refrigerator for 12-24h; centrifuge at 4500r/min for 10-20min, take the precipitate, add pure water to 100-150mL dissolve the precipitate.

In step 4 of the above method, it is preferable to add absolute ethanol to the above-mentioned supernatant to 500 mL, and let it stand in a refrigerator at 4° C. for 12 hours. Centrifuge at 4500r/min for 10min, take the precipitate, add pure water to 100mL to dissolve the precipitate.

In step 5 of the above method, add absolute ethanol to the above supernatant to 300-500 mL, and let stand in a refrigerator at 4°C for 12-24 hours. Centrifuge at 4500r/min for 10-20min, take the precipitate, add pure water to 100-150mL to dissolve the precipitate.

In step 6 of the above method, the solution can be centrifuged at 3000-5000r/min for 10-20min.

The polysaccharide extracted by the method provided by the present invention can be used for cosmetic raw materials and food industry.

Compared with the prior art, the present invention has the following outstanding substantive features:

1. The ultrasonic-assisted extraction method has the characteristics of short extraction time, low required temperature and high yield, but the ultrasonic power should not be too low or too high, otherwise the extraction rate will be low or the molecular structure and characteristics of polysaccharides will be negatively affected. The invention proposes a new extraction process, fully explores the influence of ultrasonic power on the extraction rate, overcomes the defects of the traditional ultrasonic-assisted extraction method, and realizes the rapid and efficient extraction of dendrobium polysaccharide.

2. The present invention balances many factors, such as under the premise that the number of extractions is 1, the optimal experimental parameters under the comprehensive influencing factors such as pulverization degree, solid-liquid ratio, ultrasonic time, extraction temperature, extraction time and ultrasonic power, etc. .

3. Through the treatment of this method, the extraction rate of Dendrobium officinale polysaccharide is relatively high, and the extraction rate can reach 61.12%.

4. The operation of the present invention is simple, and the obtained crude polysaccharide is relatively pure.

5. The present invention has simple operation, is easy to realize mass production of Dendrobium officinale polysaccharide, has stable operation, low energy consumption, can realize industrialized production, and the obtained crude polysaccharide is relatively pure.

6. The present invention does not introduce toxic substances, and the extracted polysaccharide can be used for cosmetic raw materials and food industries.

Detailed ways

The invention provides a novel extraction process of Dendrobium officinale polysaccharide, which is mainly to compare the ratio of material to liquid, ultrasonic time, extraction temperature, extraction time and ultrasonic power when the extraction times is 1 time, and then the extraction times are compared. research. Pulverize the dried Dendrobium officinale, the pulverization degree is: 0.1-1 mm, weigh an appropriate amount, add a certain amount of absolute ethanol, seal and soak; remove the absolute ethanol in the soaked sample, and add the sample and distilled water in an appropriate proportion to a three-necked flask ;Set the working parameters of the ultrasonic cleaner, the material-liquid ratio is 1:20-1:60 (g/mL), the ultrasonic time is 30-150min, the extraction temperature is 50-90℃, the extraction time is 120-240min and the ultrasonic power is 20%-100% (240W), stir and extract; filter the extracted solution into a conical flask; put the filtrate in an ultrasonic cleaner to ultrasonicate, and use an electric heating temperature control device for heating, and use a circulating water pump to vacuumize for reduced pressure concentration , centrifuge the concentrated solution; take the supernatant, add anhydrous ethanol for alcohol precipitation, let stand overnight, centrifuge to get the precipitate, add pure water to dissolve the precipitate; add anhydrous ethanol to the above solution for secondary alcohol precipitation, freeze Set, centrifuge to get the precipitate, add pure water to dissolve; after fully dissolving, centrifuge the solution, take the supernatant in a beaker; freeze-dry the polysaccharide solution to obtain the target Dendrobium officinale polysaccharide.

The present invention will be described in detail below with reference to the examples, but the present invention is not limited.

Embodiment 1. The extraction method of Dendrobium officinale polysaccharide

Weigh 5 g of Dendrobium officinale with a pulverization degree of 0.8-1 mm, add about 50 mL of anhydrous ethanol, seal with plastic wrap, and soak for 12 h; after removing the ethanol, add the sample to a 500 mL three-neck flask, and set the experimental parameters as follows: material-liquid ratio Water extraction was carried out under the conditions of 1:20 (g/mL), ultrasonic time 90 min, extraction temperature 70 °C, extraction time 150 min, extraction times 1 and ultrasonic power 100% (240 W); The mesh screen was coarsely filtered into a 1000mL conical flask, and the filter cake was rinsed with pure water 3 times; the filtrate was placed in an ultrasonic cleaner for ultrasound, heated with an electric heating temperature control device, and a circulating water pump was used to evacuate for decompression. Concentrate; centrifuge the concentrate (10min, 4500r/min); take the supernatant, add absolute ethanol to the above supernatant to 500mL for alcohol precipitation, and let it stand in a refrigerator at 4°C overnight; centrifuge (10min, 4500r/min) ) Take the precipitate, add pure water to 100mL to dissolve the precipitate; add absolute ethanol to the above solution to 300mL for secondary alcohol precipitation, let stand in a refrigerator at 4°C for 2h, centrifuge (10min, 4500r/min) to take the precipitate, add pure water to 150mL for dissolving; after fully dissolving, the solution was centrifuged (20min, 5000r/min), and the supernatant was taken into a beaker. The results showed that the extraction rate was 13.88%.

Embodiment 2. The extraction method of Dendrobium officinale polysaccharide

Weigh 5 g of Dendrobium officinale with a pulverization degree of 0.8-1 mm, add about 50 mL of anhydrous ethanol, seal with plastic wrap, and soak for 12 h; after removing the ethanol, add the sample to a 500 mL three-neck flask, and set the experimental parameters as follows: material-liquid ratio Water extraction was carried out under the conditions of 1:50 (g/mL), ultrasonic time 90 min, extraction temperature 70 °C, extraction time 150 min, extraction times 1 and ultrasonic power 100% (240 W); The mesh screen was coarsely filtered into a 1000mL conical flask, and the filter cake was rinsed with pure water 3 times; the filtrate was placed in an ultrasonic cleaner for ultrasound, heated with an electric heating temperature control device, and a circulating water pump was used to evacuate for decompression. Concentrate; centrifuge the concentrate (10min, 4500r/min); take the supernatant, add absolute ethanol to the above supernatant to 500mL for alcohol precipitation, and let it stand in a refrigerator at 4°C overnight; centrifuge (10min, 4500r/min) ) Take the precipitate, add pure water to 100mL to dissolve the precipitate; add absolute ethanol to the above solution to 300mL for secondary alcohol precipitation, let stand in a refrigerator at 4°C for 2h, centrifuge (10min, 4500r/min) to take the precipitate, add pure water to 150mL for dissolving; after fully dissolving, the solution was centrifuged (20min, 5000r/min), and the supernatant was taken into a beaker. The results showed that the extraction rate was 21.21%.

Embodiment 3. The extraction method of Dendrobium officinale polysaccharide

Weigh 5 g of Dendrobium officinale with a pulverization degree of 0.4-0.6 mm, add about 50 mL of absolute ethanol, seal with plastic wrap, and soak for 12 hours; after removing the ethanol, add the sample to a 500 mL three-neck flask, and set the experimental parameters as follows: feed liquid Water extraction was carried out under the conditions of a ratio of 1:50 (g/mL), ultrasonic time of 90 min, extraction temperature of 70 °C, extraction time of 150 min, extraction times of 1 time, and ultrasonic power of 100% (240 W). The 100-mesh gauze was coarsely filtered into a 1000mL conical flask, and the filter cake was rinsed three times with pure water; the filtrate was placed in an ultrasonic cleaner for ultrasound, heated with an electric heating temperature control device, and a circulating water pump was used to vacuumize the filter cake for reduction. Concentrate under pressure; centrifuge the concentrate (10 min, 4500 r/min); take the supernatant, add absolute ethanol to the above supernatant to 500 mL for alcohol precipitation, and let it stand in a refrigerator at 4°C overnight; centrifuge (10 min, 4500 r/min) min) to take the precipitate, add pure water to 100mL to dissolve the precipitate; add absolute ethanol to the above solution to 300mL for secondary alcohol precipitation, let stand in a refrigerator at 4°C for 2h, centrifuge (10min, 4500r/min) to take the precipitate, add pure water Dissolve to 150mL; after fully dissolving, centrifuge the solution (20min, 5000r/min), and take the supernatant into a beaker. The results showed that the extraction rate was 25.35%.

Embodiment 4. The extraction method of Dendrobium officinale polysaccharide

Weigh 5 g of Dendrobium officinale with a pulverization degree of 0.1-0.2 mm, add about 50 mL of absolute ethanol, seal with plastic wrap, and soak for 12 hours; after removing the ethanol, add the sample to a 500 mL three-neck flask, and set the experimental parameters as follows: feed liquid Water extraction was carried out under the conditions of a ratio of 1:50 (g/mL), ultrasonic time of 90 min, extraction temperature of 70 °C, extraction time of 150 min, extraction times of 1 time, and ultrasonic power of 100% (240 W). The 100-mesh gauze was coarsely filtered into a 1000mL conical flask, and the filter cake was rinsed three times with pure water; the filtrate was placed in an ultrasonic cleaner for ultrasound, heated with an electric heating temperature control device, and a circulating water pump was used to vacuumize the filter cake for reduction. Concentrate under pressure; centrifuge the concentrate (10 min, 4500 r/min); take the supernatant, add absolute ethanol to the above supernatant to 500 mL for alcohol precipitation, and let it stand in a refrigerator at 4°C overnight; centrifuge (10 min, 4500 r/min) min) to take the precipitate, add pure water to 100mL to dissolve the precipitate; add absolute ethanol to the above solution to 300mL for secondary alcohol precipitation, let stand in a refrigerator at 4°C for 2h, centrifuge (10min, 4500r/min) to take the precipitate, add pure water Dissolve to 150mL; after fully dissolving, centrifuge the solution (20min, 5000r/min), and take the supernatant into a beaker. The results showed that the extraction rate was 34.43%.

Embodiment 5. The extraction method of Dendrobium officinale polysaccharide

Weigh 5 g of Dendrobium officinale with a pulverization degree of 0.1-0.2 mm, add about 50 mL of absolute ethanol, seal with plastic wrap, and soak for 12 hours; after removing the ethanol, add the sample to a 500 mL three-neck flask, and set the experimental parameters as follows: feed liquid Water extraction was carried out under the conditions of a ratio of 1:50 (g/mL), ultrasonic time of 30 min, extraction temperature of 70 °C, extraction time of 150 min, extraction times of 1 time, and ultrasonic power of 100% (240 W). The 100-mesh gauze was coarsely filtered into a 1000mL conical flask, and the filter cake was rinsed three times with pure water; the filtrate was placed in an ultrasonic cleaner for ultrasound, heated with an electric heating temperature control device, and a circulating water pump was used to vacuumize the filter cake for reduction. Concentrate under pressure; centrifuge the concentrate (10 min, 4500 r/min); take the supernatant, add absolute ethanol to the above supernatant to 500 mL for alcohol precipitation, and let it stand in a refrigerator at 4°C overnight; centrifuge (10 min, 4500 r/min) min) to take the precipitate, add pure water to 100mL to dissolve the precipitate; add absolute ethanol to the above solution to 300mL for secondary alcohol precipitation, let stand in a refrigerator at 4°C for 2h, centrifuge (10min, 4500r/min) to take the precipitate, add pure water Dissolve to 150mL; after fully dissolving, centrifuge the solution (20min, 5000r/min), and take the supernatant into a beaker. The results showed that the extraction rate was 29.56%.

Embodiment 6. The extraction method of Dendrobium officinale polysaccharide

Weigh 5 g of Dendrobium officinale with a pulverization degree of 0.1-0.2 mm, add about 50 mL of absolute ethanol, seal with plastic wrap, and soak for 12 hours; after removing the ethanol, add the sample to a 500 mL three-neck flask, and set the experimental parameters as follows: feed liquid Water extraction was carried out under the conditions of a ratio of 1:50 (g/mL), ultrasonic time of 120 min, extraction temperature of 70 °C, extraction time of 150 min, extraction times of 1 time, and ultrasonic power of 100% (240 W). The 100-mesh gauze was coarsely filtered into a 1000mL conical flask, and the filter cake was rinsed three times with pure water; the filtrate was placed in an ultrasonic cleaner for ultrasound, heated with an electric heating temperature control device, and a circulating water pump was used to vacuumize the filter cake for reduction. Concentrate under pressure; centrifuge the concentrate (10 min, 4500 r/min); take the supernatant, add absolute ethanol to the above supernatant to 500 mL for alcohol precipitation, and let it stand in a refrigerator at 4°C overnight; centrifuge (10 min, 4500 r/min) min) to take the precipitate, add pure water to 100mL to dissolve the precipitate; add absolute ethanol to the above solution to 300mL for secondary alcohol precipitation, let stand in a refrigerator at 4°C for 2h, centrifuge (10min, 4500r/min) to take the precipitate, add pure water Dissolve to 150mL; after fully dissolving, centrifuge the solution (20min, 5000r/min), and take the supernatant into a beaker. The results showed that the extraction rate was 36.70%.

Embodiment 7. The extraction method of Dendrobium officinale polysaccharide

Weigh 5 g of Dendrobium officinale with a pulverization degree of 0.1-0.2 mm, add about 50 mL of absolute ethanol, seal with plastic wrap, and soak for 12 hours; after removing the ethanol, add the sample to a 500 mL three-neck flask, and set the experimental parameters as follows: feed liquid Water extraction was carried out under the conditions of a ratio of 1:50 (g/mL), ultrasonic time of 120 min, extraction temperature of 70 °C, extraction time of 150 min, extraction times of 1 time, and ultrasonic power of 20% (240 W). The 100-mesh gauze was coarsely filtered into a 1000mL conical flask, and the filter cake was rinsed three times with pure water; the filtrate was placed in an ultrasonic cleaner for ultrasound, heated with an electric heating temperature control device, and a circulating water pump was used to vacuumize the filter cake for reduction. Concentrate under pressure; centrifuge the concentrate (10 min, 4500 r/min); take the supernatant, add absolute ethanol to the above supernatant to 500 mL for alcohol precipitation, and let it stand in a refrigerator at 4°C overnight; centrifuge (10 min, 4500 r/min) min) to take the precipitate, add pure water to 100mL to dissolve the precipitate; add absolute ethanol to the above solution to 300mL for secondary alcohol precipitation, let stand in a refrigerator at 4°C for 2h, centrifuge (10min, 4500r/min) to take the precipitate, add pure water Dissolve to 150mL; after fully dissolving, centrifuge the solution (20min, 5000r/min), and take the supernatant into a beaker. The results showed that the extraction rate was 37.63%.

Embodiment 8. The extraction method of Dendrobium officinale polysaccharide

Weigh 5 g of Dendrobium officinale with a pulverization degree of 0.1-0.2 mm, add about 50 mL of absolute ethanol, seal with plastic wrap, and soak for 12 hours; after removing the ethanol, add the sample to a 500 mL three-neck flask, and set the experimental parameters as follows: feed liquid Water extraction was carried out under the conditions of a ratio of 1:50 (g/mL), ultrasonic time of 120 min, extraction temperature of 70 °C, extraction time of 150 min, extraction times of 1 time and ultrasonic power of 60% (240 W). The 100-mesh gauze was coarsely filtered into a 1000mL conical flask, and the filter cake was rinsed three times with pure water; the filtrate was placed in an ultrasonic cleaner for ultrasound, heated with an electric heating temperature control device, and a circulating water pump was used to vacuumize the filter cake for reduction. Concentrate under pressure; centrifuge the concentrate (10 min, 4500 r/min); take the supernatant, add absolute ethanol to the above supernatant to 500 mL for alcohol precipitation, and let it stand in a refrigerator at 4°C overnight; centrifuge (10 min, 4500 r/min) min) to take the precipitate, add pure water to 100mL to dissolve the precipitate; add absolute ethanol to the above solution to 300mL for secondary alcohol precipitation, let stand in a refrigerator at 4°C for 2h, centrifuge (10min, 4500r/min) to take the precipitate, add pure water Dissolve to 150mL; after fully dissolving, centrifuge the solution (20min, 5000r/min), and take the supernatant into a beaker. The results showed that the extraction rate was 41.47%.

Embodiment 9. The extraction method of Dendrobium officinale polysaccharide

Weigh 5 g of Dendrobium officinale with a pulverization degree of 0.1-0.2 mm, add about 50 mL of absolute ethanol, seal with plastic wrap, and soak for 12 hours; after removing the ethanol, add the sample to a 500 mL three-neck flask, and set the experimental parameters as follows: feed liquid Water extraction was carried out under the conditions of a ratio of 1:50 (g/mL), ultrasonic time of 120 min, extraction temperature of 50 °C, extraction time of 150 min, extraction times of 1 time, and ultrasonic power of 60% (240 W). The 100-mesh gauze was coarsely filtered into a 1000mL conical flask, and the filter cake was rinsed three times with pure water; the filtrate was placed in an ultrasonic cleaner for ultrasound, heated with an electric heating temperature control device, and a circulating water pump was used to vacuumize the filter cake for reduction. Concentrate under pressure; centrifuge the concentrate (10 min, 4500 r/min); take the supernatant, add absolute ethanol to the above supernatant to 500 mL for alcohol precipitation, and let it stand in a refrigerator at 4°C overnight; centrifuge (10 min, 4500 r/min) min) to take the precipitate, add pure water to 100mL to dissolve the precipitate; add absolute ethanol to the above solution to 300mL for secondary alcohol precipitation, let stand in a refrigerator at 4°C for 2h, centrifuge (10min, 4500r/min) to take the precipitate, add pure water Dissolve to 150mL; after fully dissolving, centrifuge the solution (20min, 5000r/min), and take the supernatant into a beaker. The results showed that the extraction rate was 38.48%.

Embodiment 10. The extraction method of Dendrobium officinale polysaccharide

Weigh 5 g of Dendrobium officinale with a pulverization degree of 0.1-0.2 mm, add about 50 mL of absolute ethanol, seal with plastic wrap, and soak for 12 hours; after removing the ethanol, add the sample to a 500 mL three-neck flask, and set the experimental parameters as follows: feed liquid Water extraction was carried out under the conditions of a ratio of 1:50 (g/mL), ultrasonic time of 120 min, extraction temperature of 80 °C, extraction time of 150 min, extraction times of 1 time, and ultrasonic power of 60% (240 W). The 100-mesh gauze was coarsely filtered into a 1000mL conical flask, and the filter cake was rinsed three times with pure water; the filtrate was placed in an ultrasonic cleaner for ultrasound, heated with an electric heating temperature control device, and a circulating water pump was used to vacuumize the filter cake for reduction. Concentrate under pressure; centrifuge the concentrate (10 min, 4500 r/min); take the supernatant, add absolute ethanol to the above supernatant to 500 mL for alcohol precipitation, and let it stand in a refrigerator at 4°C overnight; centrifuge (10 min, 4500 r/min) min) to take the precipitate, add pure water to 100mL to dissolve the precipitate; add absolute ethanol to the above solution to 300mL for secondary alcohol precipitation, let stand in a refrigerator at 4°C for 2h, centrifuge (10min, 4500r/min) to take the precipitate, add pure water Dissolve to 150mL; after fully dissolving, centrifuge the solution (20min, 5000r/min), and take the supernatant into a beaker. The results showed that the extraction rate was 44.64%.

Embodiment 11. The extraction method of Dendrobium officinale polysaccharide

Weigh 5 g of Dendrobium officinale with a pulverization degree of 0.1-0.2 mm, add about 50 mL of absolute ethanol, seal with plastic wrap, and soak for 12 hours; after removing the ethanol, add the sample to a 500 mL three-neck flask, and set the experimental parameters as follows: feed liquid Water extraction was carried out under the conditions of a ratio of 1:50 (g/mL), ultrasonic time of 120 min, extraction temperature of 80 °C, extraction time of 120 min, extraction times of 1 time, and ultrasonic power of 60% (240 W). The 100-mesh gauze was coarsely filtered into a 1000mL conical flask, and the filter cake was rinsed three times with pure water; the filtrate was placed in an ultrasonic cleaner for ultrasound, heated with an electric heating temperature control device, and a circulating water pump was used to vacuumize the filter cake for reduction. Concentrate under pressure; centrifuge the concentrate (10 min, 4500 r/min); take the supernatant, add absolute ethanol to the above supernatant to 500 mL for alcohol precipitation, and let it stand in a refrigerator at 4°C overnight; centrifuge (10 min, 4500 r/min) min) to take the precipitate, add pure water to 100mL to dissolve the precipitate; add absolute ethanol to the above solution to 300mL for secondary alcohol precipitation, let stand in a refrigerator at 4°C for 2h, centrifuge (10min, 4500r/min) to take the precipitate, add pure water Dissolve to 150mL; after fully dissolving, centrifuge the solution (20min, 5000r/min), and take the supernatant into a beaker. The results showed that the extraction rate was 39.50%.

Embodiment 12. The extraction method of Dendrobium officinale polysaccharide

Weigh 5 g of Dendrobium officinale with a pulverization degree of 0.1-0.2 mm, add about 50 mL of absolute ethanol, seal with plastic wrap, and soak for 12 hours; after removing the ethanol, add the sample to a 500 mL three-neck flask, and set the experimental parameters as follows: feed liquid Water extraction was carried out under the conditions of a ratio of 1:50 (g/mL), ultrasonic time of 120 min, extraction temperature of 80 °C, extraction time of 210 min, extraction times of 1 time, and ultrasonic power of 60% (240 W). The 100-mesh gauze was coarsely filtered into a 1000mL conical flask, and the filter cake was rinsed three times with pure water; the filtrate was placed in an ultrasonic cleaner for ultrasound, heated with an electric heating temperature control device, and a circulating water pump was used to vacuumize the filter cake for reduction. Concentrate under pressure; centrifuge the concentrate (10 min, 4500 r/min); take the supernatant, add absolute ethanol to the above supernatant to 500 mL for alcohol precipitation, and let it stand in a refrigerator at 4°C overnight; centrifuge (10 min, 4500 r/min) min) to take the precipitate, add pure water to 100mL to dissolve the precipitate; add absolute ethanol to the above solution to 300mL for secondary alcohol precipitation, let stand in a refrigerator at 4°C for 2h, centrifuge (10min, 4500r/min) to take the precipitate, add pure water Dissolve to 150mL; after fully dissolving, centrifuge the solution (20min, 5000r/min), and take the supernatant into a beaker. The results showed that the extraction rate was 51.48%.

Embodiment 13. The extraction method of Dendrobium officinale polysaccharide

Weigh 5 g of Dendrobium officinale with a pulverization degree of 0.1-0.2 mm, add about 50 mL of absolute ethanol, seal with plastic wrap, and soak for 12 hours; after removing the ethanol, add the sample to a 500 mL three-neck flask, and set the experimental parameters as follows: feed liquid Water extraction was carried out under the conditions of a ratio of 1:50 (g/mL), ultrasonic time of 120 min, extraction temperature of 80 °C, extraction time of 210 min, extraction times of 3 times and ultrasonic power of 60% (240 W). The 100-mesh gauze was coarsely filtered into a 1000mL conical flask, and the filter cake was rinsed three times with pure water; the filtrate was placed in an ultrasonic cleaner for ultrasound, heated with an electric heating temperature control device, and a circulating water pump was used to vacuumize the filter cake for reduction. Concentrate under pressure; centrifuge the concentrate (10 min, 4500 r/min); take the supernatant, add absolute ethanol to the above supernatant to 500 mL for alcohol precipitation, and let it stand in a refrigerator at 4°C overnight; centrifuge (10 min, 4500 r/min) min) to take the precipitate, add pure water to 100mL to dissolve the precipitate; add absolute ethanol to the above solution to 300mL for secondary alcohol precipitation, let stand in a refrigerator at 4°C for 2h, centrifuge (10min, 4500r/min) to take the precipitate, add pure water Dissolve to 150mL; after fully dissolving, centrifuge the solution (20min, 5000r/min), and take the supernatant into a beaker. The results showed that the extraction rate was 61.12%.

The method provided by the above embodiment has the following characteristics:

1. The extraction method of Dendrobium officinale polysaccharide provided by the present invention is a new type of extraction process utilizing ultrasonic wave, and fully explores the factors affecting the extraction rate, overcomes the defects of the traditional ultrasonic-assisted extraction method, and realizes Dendrobium polysaccharide. Fast and efficient extraction.

2. The present invention balances many factors, such as under the premise that the number of extractions is 1, the optimal experimental parameters under the comprehensive influencing factors such as pulverization degree, solid-liquid ratio, ultrasonic time, extraction temperature, extraction time and ultrasonic power, etc. .

3. Through the treatment of this method, the yield of Dendrobium officinale polysaccharide is high, up to 61.12%.

4. The present invention has simple operation, is easy to realize mass production of Dendrobium officinale polysaccharide, has stable operation, low energy consumption, can realize industrialized production, and the obtained crude polysaccharide is relatively pure.

5. The present invention does not introduce toxic substances, and the extracted polysaccharide can be used for cosmetic raw materials and food industries.

Claims (10)

1. The extraction method of the dendrobium candidum polysaccharide is characterized in that absolute ethyl alcohol is added into dried and crushed dendrobium candidum, and the mixture is sealed and soaked; removing ethanol, adding distilled water, performing ultrasonic treatment, extracting, concentrating the obtained polysaccharide solution under reduced pressure, and removing impurities with anhydrous ethanol.

2. The extraction method of the dendrobium officinale polysaccharide as claimed in claim 1, wherein the granularity of the dendrobium officinale is 0.1-0.2mm, 4.8-5.2g of the dendrobium officinale is weighed, 48-52mL of absolute ethyl alcohol is added, and the mixture is sealed by a preservative film and soaked for 11-13 h.

3. The extraction method of dendrobium officinale polysaccharide as claimed in claim 1, which is characterized by comprising the following steps:

step 1, removing absolute ethyl alcohol in the soaked sample, and adding the sample and distilled water into a three-neck flask according to a proper proportion;

step 2, setting working parameters of an ultrasonic cleaner, and stirring and extracting;

step 3, filtering the extracted solution into a conical flask, putting the filtrate into an ultrasonic cleaner for ultrasonic treatment, heating by using an electric heating temperature control device, simultaneously performing vacuum pumping by using a circulating water pump for reduced pressure concentration, and centrifuging the concentrated solution;

step 4, adding absolute ethyl alcohol into the supernatant for alcohol precipitation, standing overnight, centrifuging to obtain a precipitate, and adding pure water to dissolve the precipitate;

step 5, adding absolute ethyl alcohol into the solution for secondary alcohol precipitation, standing in a refrigerator, centrifuging to obtain precipitate, and adding pure water for dissolving;

step 6, after full dissolution, centrifuging the solution, and taking supernatant liquid in a beaker;

and 7, carrying out freeze drying on the polysaccharide solution to obtain the target dendrobium officinale polysaccharide.

4. The method according to claim 3, wherein the ratio of the sample to distilled water in step 1 is 1: 20-1: 60g/mL, and charged into a 500mL three-necked flask.

5. The method of claim 3, wherein the 240W ultrasonic cleaner parameters are set in step 2 to: heating at 50-90 deg.C for 120-.

6. The method according to claim 3, wherein the extracted solution is coarsely filtered with a 100 mesh screen into a 1000mL conical flask in step 3, and the filter cake is washed several times with pure water.

7. The method as claimed in claim 3, wherein the concentrate is centrifuged at 3000-4500r/min for 10-20min in step 3.

8. The method as claimed in claim 3, wherein the absolute ethanol is added to the supernatant in step 4 to 500mL, and the mixture is left standing in a refrigerator at 4 ℃ for 12-24 h; centrifuging at 4500r/min for 10-20min, collecting precipitate, adding pure water to 100-.

9. The method according to claim 3, wherein in step 5, absolute ethanol is added for secondary alcohol precipitation, and the mixture is kept still for 2 to 8 hours at 4 ℃ in a refrigerator; centrifuging at 3000-4500r/min for 10-20min, collecting the precipitate, adding pure water to 150-200mL for dissolution.

10. The method according to claim 3, wherein the solution is centrifuged at 3000-5000r/min for 10-20min in step 6.