Method for simultaneously extracting rhizoma drynariae starch and flavone

Abstract

The invention provides a method for simultaneously extracting rhizoma drynariae starch and flavone, and belongs to the field of traditional Chinese medicine processing and active ingredient extraction. According to the method disclosed by the invention, by exploring extraction process conditions such as an extraction solvent, a material-to-liquid ratio, an extraction temperature, a starch drying temperature and the like, the utilization rate of the raw materials is effectively improved, the whiteness of the starch and the extraction rate of flavone are increased, and the raw materials are provided for development of edible rhizoma drynariae starch and development of related health-care foods containing active ingredients; the extraction efficiency and the economic value of the rhizoma drynariae are improved.

Description

Technical Field

[0001] The invention belongs to the field of traditional Chinese medicine processing and active component extraction, and particularly relates to a method for simultaneously extracting rhizoma drynariae starch and flavonoids. Background Art

[0002] Drynaria fortunei is derived from the plant Quercus serrata of the Polypodiaceae family. Drynaria fortunei The dried rhizome of Kunze J.Sm. is a blood-activating and stasis-removing medicine, which is used in the commonly used compound preparations such as Qianggu Capsule, Jiegu Qili Tablets, and Zhitong Zijin Pills. It has the effects of healing wounds and relieving pain, tonifying the kidney and strengthening bones; it can also be used externally to eliminate wind and remove spots. Modern research has shown that Drynaria can increase blood calcium and phosphorus levels, promote fracture healing, and resist inflammation and atherosclerosis.

[0003] The chemical components of Drynaria Rhizoma include flavonoids, triterpenes, lignin, phenylpropanoids and other compounds, and flavonoids are its main active substances. Flavonoids have multiple biological activities such as anti-inflammatory, antibacterial and antibacterial, anti-tumor, antioxidant, anti-aging, anti-fatigue, lowering blood lipids, lowering serum cholesterol, and treating cardiovascular diseases. Therefore, the extraction of total flavonoids and the study of their activity have attracted widespread attention. At present, the existing processes mainly use ultrasound, reflux, etc. to extract the dry rhizomes of Drynaria Rhizoma (Wei Guolan, Long Jiefeng, Bai Feng. Study on the process of ultrasonic-assisted extraction of total flavonoids from Drynaria Rhizoma [J]. Guangzhou Chemical Industry, 2022, 50(7): 42-44. DOI: 10.3969 / j.issn.1001-9677.2022.07.014.), but the extraction effect is not good, and the total flavonoid extraction rate is only 1.04%-7.53%.

[0004] Drynaria is widely used in traditional Chinese medicine, but there are few products developed as health food. Existing drynaria patents are mainly concentrated in the development of preparations and the production technology of Chinese herbal medicines. With the country's emphasis on the health industry and traditional Chinese medicine, the demand for functional ingredients of traditional Chinese medicine has also increased accordingly. Exploring the rapid and efficient extraction of starch and flavonoids from drynaria has become a problem that needs to be solved in the future. Summary of the invention

[0005] The present invention proposes a method for simultaneously extracting drynaria starch and flavonoids. By optimizing various parameters and steps such as extraction solvent, solid-liquid ratio, extraction temperature, starch drying temperature, extraction process, etc., the utilization rate of raw materials is effectively improved, the whiteness of starch and the extraction rate of flavonoids are increased. Compared with the traditional extraction of flavonoid effective ingredients, starch as a new product is added, and a new direction of developing drynaria into a health food is added, which lays a foundation for the deep processing of related products in the future and broadens the utilization value of drynaria.

[0006] The present invention provides a method for simultaneously extracting rhizoma drynariae starch and flavonoids, comprising the following steps: S1. Take fresh rhizomes of Drynariae Rhizoma, soak them, remove impurities, remove the epidermis, cut them into small pieces, add ethanol solution, beat and filter, and obtain filtrate and residue; S2. The filtrate obtained in S1 is allowed to stand to obtain a supernatant and a precipitate. The supernatant is evaporated to dryness to obtain Rhizoma Drynariae flavonoids. The precipitate is repeatedly added with water and centrifuged to remove the colored layer on the surface. The Rhizoma Drynariae starch is obtained by drying.

[0007] Furthermore, the volume fraction of the ethanol solution in S1 is 50%.

[0008] Furthermore, the drynaria rhizome in S1 was divided into small pieces and added to the ethanol solution at a solid-liquid ratio of 1:5, g / mL.

[0009] Furthermore, the beating time in S1 is 5 min.

[0010] Furthermore, the filtration in S1 uses a 100-200 mesh sieve.

[0011] Furthermore, the centrifugation speed in S2 is 5000 rpm and the time is 20 min.

[0012] Furthermore, the drying temperature in S2 is 40-50°C.

[0013] The present invention simultaneously extracts starch and flavonoid components of Drynariae Rhizoma. Starch is a high molecular carbohydrate, which is insoluble in cold water and ethanol solution, while flavonoid components are mostly present in the form of flavonoid glycosides in plants, which have high solubility in strong polar solvents such as water and ethanol. The present invention uses a wall-breaking technology to free starch and flavonoid components present in Drynariae Rhizoma tissues, sieves to remove tissue fragments, obtains an extract, and centrifuges to precipitate starch insoluble in water and ethanol in the precipitate, and flavonoid components are present in the supernatant. Since the extraction solvent, solid-liquid ratio, extraction temperature and centrifugal parameter settings all have an impact on the extraction rate, starch whiteness and extraction efficiency, the present invention optimizes these parameters, and simultaneously investigates the starch drying temperature to prevent starch gelatinization.

[0014] Compared with the prior art, the present invention has the following beneficial effects: The invention provides a method for simultaneously extracting drynaria starch and flavonoids. The selected raw materials in the method do not need excessive processing. The fresh rhizomes of drynaria are directly pulped and filtered. By exploring extraction process conditions such as extraction solvent, material-liquid ratio, extraction temperature, starch drying temperature, etc., the utilization rate of raw materials is effectively improved, the whiteness of starch and the extraction rate of flavonoids are increased, and the extraction effect is better than that of existing ultrasonic, reflux and other processes. Raw materials are provided for the development of edible starch of drynaria and the development of related health foods containing active ingredients, and the extraction efficiency and economic value of drynaria are increased. DETAILED DESCRIPTION

[0015] To make the purpose, technical scheme and advantages of the present invention clearer, the technical scheme of the present invention will be clearly and completely described below in conjunction with the embodiments. If the specific conditions are not specified in the embodiments, they are carried out according to the normal conditions or the conditions recommended by the manufacturer. If the reagents or instruments used are not specified by the manufacturer, they are all conventional products that can be purchased commercially. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments. The following description of at least one exemplary embodiment is actually only illustrative and is by no means intended to limit the present application and its application or use. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without making creative work are within the scope of protection of the present application.

[0016] Example 1 A method for simultaneously extracting rhizoma drynariae starch and flavonoids comprises the following steps: 1. Dig up fresh rhizomes of Drynaria fortunei, soak them in clean water for 3 h, select the thick and plump rhizomes, remove impurities such as moss and bark on the surface, scrape off the epidermis, cut the Drynaria fortunei into small pieces of 1-2 cm while it is still fresh, add 50% ethanol solution by volume (solid-liquid ratio 1:5, g / mL), beat it into a pulp with a wall breaking machine (5 min), and filter it through a 100-mesh sieve to obtain the filtrate and residue.

[0017] 2. Take the filtrate and let it stand overnight to obtain the supernatant and precipitate. Concentrate the supernatant and evaporate to dryness to obtain flavonoids. Add water to the precipitate and centrifuge (5000 rpm, 20 min), remove the supernatant after centrifugation, and scrape off the colored layer on the surface of the precipitate; continue to add water and repeat the operation three times, scrape out the remaining white precipitate and place it on a tray, dry it in an oven at 40°C, and crush it after drying to obtain Drynaria starch.

[0018] Example 2 A method for simultaneously extracting rhizoma drynariae starch and flavonoids comprises the following steps: 1. Dig up fresh rhizomes of Drynaria fortunei, soak them in clean water for 3 h, select the thick and plump rhizomes, remove impurities such as moss and bark on the surface, scrape off the epidermis, cut the Drynaria fortunei into small pieces of 1-2 cm while it is still fresh, add 50% ethanol solution by volume (solid-liquid ratio 1:5, g / mL), beat it into a pulp with a wall breaking machine (5 min), and filter it through a 200-mesh sieve to obtain the filtrate and residue.

[0019] 2. Take the filtrate and let it stand overnight to obtain the supernatant and precipitate. Concentrate the supernatant and evaporate to dryness to obtain flavonoids. Add water to the precipitate and centrifuge (5000 rpm, 20 min), remove the supernatant after centrifugation, and scrape off the colored layer on the surface of the precipitate; continue to add water and repeat the operation three times, scrape out the remaining white precipitate and place it on a tray, dry it in an oven at 40°C, and crush it after drying to obtain Drynaria starch.

[0020] Example 3 A method for simultaneously extracting rhizoma drynariae starch and flavonoids comprises the following steps: 1. Dig up fresh rhizomes of Drynaria fortunei, soak them in clean water for 3 h, select the thick and plump rhizomes, remove impurities such as moss and bark on the surface, scrape off the epidermis, cut the Drynaria fortunei into small pieces of 1-2 cm while it is still fresh, add 50% ethanol solution by volume (solid-liquid ratio 1:5, g / mL), beat it into a pulp with a wall breaking machine (5 min), and filter it through a 100-mesh sieve to obtain the filtrate and residue.

[0021] 2. Take the filtrate and let it stand overnight to obtain the supernatant and precipitate. Concentrate the supernatant and evaporate to dryness to obtain flavonoids. Add water to the precipitate and centrifuge (5000 rpm, 20 min), remove the supernatant after centrifugation, and scrape off the colored layer on the surface of the precipitate; continue to add water and repeat the operation three times, scrape out the remaining white precipitate and place it on a tray, dry it in an oven at 45°C, and crush it after drying to obtain Drynaria starch.

[0022] Example 4 A method for simultaneously extracting rhizoma drynariae starch and flavonoids comprises the following steps: 1. Dig up fresh rhizomes of Drynaria fortunei, soak them in clean water for 3 h, select the thick and plump rhizomes, remove impurities such as moss and bark on the surface, scrape off the epidermis, cut the Drynaria fortunei into small pieces of 1-2 cm while it is still fresh, add 50% ethanol solution by volume (solid-liquid ratio 1:5, g / mL), beat it into a pulp with a wall breaking machine (5 min), and filter it through a 100-mesh sieve to obtain the filtrate and residue.

[0023] 2. Take the filtrate and let it stand overnight to obtain the supernatant and precipitate. Concentrate the supernatant and evaporate to dryness to obtain flavonoids. Add water to the precipitate and centrifuge (5000 rpm, 20 min), remove the supernatant after centrifugation, and scrape off the colored layer on the surface of the precipitate; continue to add water and repeat the operation three times, scrape out the remaining white precipitate and place it on a tray, dry it in an oven at 50°C, and crush it after drying to obtain Drynaria starch.

[0024] Comparative Example 1 Compared with Example 1, the only difference is that in step 1, the solid-liquid ratio of Rhizoma Drynariae to 50% ethanol solution is 1:3 (g / mL), and the conditions of other steps are the same.

[0025] Comparative Example 2 Compared with Example 1, the only difference is that in step 1, the solid-liquid ratio of Rhizoma Drynariae to 50% ethanol solution is 1:10 (g / mL), and the conditions of other steps are the same.

[0026] Comparative Example 3 Compared with Example 1, the only difference is that in step 1, the solid-liquid ratio of Rhizoma Drynariae to 50% ethanol solution is 1:15 (g / mL), and the conditions of other steps are the same.

[0027] Comparative Example 4 Compared with Example 1, the only difference is that the starch drying temperature in step 2 is 55° C., and the conditions in other steps are the same.

[0028] Comparative Example 5 Compared with Example 1, the only difference is that pure water is used as the solvent instead of 50% ethanol solution in step 1, and the conditions of other steps are the same.

[0029] Comparative Example 6 Compared with Example 1, the only difference is that in step 1, 30% ethanol solution is used as the solvent instead of 50% ethanol solution, and the conditions of other steps are the same.

[0030] Comparative Example 7 Compared with Example 1, the only difference is that in step 1, 70% ethanol solution is used as the solvent instead of 50% ethanol solution, and the conditions of other steps are the same.

[0031] Comparative Example 8 Compared with Example 1, the only difference is that in step 1, 95% ethanol solution is used as the solvent instead of 50% ethanol solution, and the conditions of other steps are the same.

[0032] Comparative Example 9 Compared with Example 1, the only difference is that the grinding and beating time of the wall breaking machine in step 1 is 2 min, and the conditions of other steps are the same.

[0033] Comparative Example 10 Compared with Example 1, the only difference is that the grinding and beating time of the wall breaking machine in step 1 is 10 min, and the conditions of other steps are the same.

[0034] Comparative Example 11 Compared with Example 1, the only difference is that in step 2, the centrifugal speed is 1000 rpm, the centrifugal time is 30 min, and the other step conditions are the same.

[0035] Comparative Example 12 Compared with Example 1, the only difference is that in step 2, the centrifugal speed is 5000 rpm, the centrifugal time is 10 min, and the other step conditions are the same.

[0036] Comparative Example 13 Compared with Example 1, the only difference is that in step 2, the centrifugal speed is 8000 rpm, the centrifugal time is 10 min, and the conditions of other steps are the same.

[0037] The selection of extraction solvents, solid-liquid ratio of Rhizoma Drynariae to solvent, extraction process parameters, and starch drying temperature for each embodiment and comparative example are shown in the following table.

[0038] Table 1

[0039] The flavonoids, starch extraction rate and starch whiteness obtained by the above examples and comparative examples were tested; the flavonoid content was determined by colorimetry, rutin was used as a standard, and the starch whiteness was measured by a colorimeter. The comparison results are shown in Table 2.

[0040] Determination of total flavonoids content: Preparation of reference solution Take 50 mg of rutin reference solution, weigh accurately, place in a 25 mL volumetric flask, add appropriate amount of ethanol, place in a water bath and heat slightly to dissolve, cool, add ethanol to the mark, shake well. Accurately measure 10 mL, place in a 100 mL volumetric flask, add water to the mark, shake well, and obtain (each 1 mL contains 0.2 mg of rutin).

[0041] Preparation of standard curve: Accurately measure 1 mL, 2 mL, 3 mL, 4 mL, 5 mL and 6 mL of the reference solution, place them in 25 mL volumetric flasks respectively, add water to 6.0 mL each, add 1 mL of 5% sodium nitrite solution, mix well, let stand for 6 minutes, add 1 mL of 10% aluminum nitrate solution, shake well, let stand for 6 minutes, add 10 mL of sodium hydroxide test solution, add water to the scale, shake well, let stand for 15 minutes, use the corresponding reagent as blank, measure the absorbance at a wavelength of 500 nm according to the UV-Vis spectrophotometry, and draw the standard curve with absorbance as the ordinate and concentration as the abscissa.

[0042] Determination method: After the supernatant of Drynaria fortunei after centrifugation is fixed to volume, accurately measure 0.5 mL and place it in a 25 mL volumetric flask. According to the method under the standard curve preparation, from "add water to 6.0 mL", determine the absorbance according to the law, read the weight of rutin in the test solution from the standard curve, and calculate it.

[0043] Starch whiteness determination: The whiteness meter is used for measurement. After the sample is fully mixed, it is loaded into the sample box provided by the whiteness meter, and a sample white plate is made according to the method specified by the whiteness meter. The measurement wavelength is 457 nm. Calibrate with a standard white plate made of high-grade pure magnesium oxide marked with whiteness, measure the sample white plate with a whiteness meter, read the whiteness value, and the whiteness is expressed as the sample whiteness value measured by the whiteness meter. Parallel measurements are performed 3 times, and the arithmetic mean of the parallel experiments is taken as the result, retaining one decimal place. The absolute difference of the experimental results should not exceed 0.2 at ordinary times. If it exceeds, re-measure.

[0044] Table 2

[0045] As shown in Table 2, the starch and flavonoids extracted from Rhizoma Drynariae by the process methods of Examples 1-4 have a starch extraction rate of more than 12%, a total flavonoid extraction rate of more than 10%, and a starch whiteness of more than 56. In particular, the starch extraction rate of Example 1 reaches 12.91%, and the flavonoid extraction rate reaches 11.24%.

[0046] Compared with Example 1, Comparative Examples 1, 2 and 3 investigated the effects of different liquid-to-solid ratios on the extraction rates of starch and flavonoids and starch whiteness. The results showed that different liquid-to-solid ratios had little effect on starch whiteness. When the liquid-to-solid ratio was 3:1, the extraction was incomplete, and the extraction rates of total flavonoids and starch were less than those of Example 1. When the liquid-to-liquid ratio was 10:1 and 15:1, since the flavonoids and starch were completely extracted at the liquid-to-solid ratio of 5:1, the extraction rate no longer increased with increasing the liquid-to-solid ratio. However, the increase in the extraction solvent and the subsequent concentration time would lead to an increase in the investment cost. Therefore, the liquid-to-liquid ratio was controlled at 1:5 in the present invention.

[0047] Compared with Example 1, Example 3 and Example 4, the starch drying temperature in Comparative Example 4 is 55°C, the whiteness of the obtained starch decreases, and there are signs of gelatinization during the drying process, so the starch drying temperature should be controlled at 40-50°C.

[0048] Compared with Example 1, Comparative Examples 5-8 investigated the extraction solvent. When the ethanol concentration was less than 50%, the total flavonoid extraction rate increased with the increase of ethanol concentration. When the ethanol concentration was greater than 50%, the total flavonoid extraction rate decreased with the increase of ethanol concentration. Free flavonoids have a large solubility in ethanol, and flavonoid glycosides have a large solubility in water and low concentrations of ethanol. When the ethanol concentration was 50%, the total amount of free flavonoids and flavonoid glycosides reached the maximum.

[0049] Compared with Example 1, the extraction time of Comparative Example 9 was shortened to 2 min. Although the extraction efficiency was increased, the total flavonoids and starch extraction rates were reduced due to insufficient extraction. In Comparative Example 10, the extraction time was increased to 10 min, and the whiteness of the extracted starch decreased, but the extraction rates of starch and flavonoids did not increase significantly. It is speculated that the extraction time was too long, which increased the oxidation probability, causing the color of the extract to darken and affecting the whiteness of the starch. The extraction time of the present invention was controlled at 5 min.

[0050] Compared with Example 1, Comparative Examples 11-13 investigated the effects of the centrifugal speed and centrifugal time on the starch extraction rate and whiteness. When the speed was too low or the centrifugal time was insufficient, the starch sedimentation was incomplete and the starch extraction rate was slightly reduced. When the speed was too high, the starch and impurities could not be completely separated, resulting in more impurities in the obtained starch and reduced whiteness. The present invention finally controlled the centrifugal speed at 5000 rpm and the centrifugal time at 20 min.

[0051] Finally, it should be emphasized that the above is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various changes and modifications. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A method for simultaneously extracting starch and flavonoids from Rhizoma Drynariae, characterized in that: The steps include: S1. Take fresh rhizomes of Drynariae Rhizoma, soak them, remove impurities, remove the epidermis, cut them into small pieces, add ethanol solution, beat and filter, and obtain filtrate and residue; S2. The filtrate obtained in S1 is allowed to stand to obtain a supernatant and a precipitate. The supernatant is evaporated to dryness to obtain Rhizoma Drynariae flavonoids. The precipitate is repeatedly added with water and centrifuged to remove the colored layer on the surface. The Rhizoma Drynariae starch is obtained by drying.

2. The method for simultaneously extracting Rhizoma Drynariae starch and flavonoids according to claim 1, characterized in that: The volume fraction of the ethanol solution in S1 is 50%.

3. The method for simultaneously extracting Rhizoma Drynariae starch and flavonoids according to claim 1, characterized in that: In S1, the solid-liquid ratio of the Rhizoma Drynariae to the ethanol solution after being cut into small pieces was 1:5, g / mL.

4. The method for simultaneously extracting Rhizoma Drynariae starch and flavonoids according to claim 1, characterized in that: The beating time described in S1 is 5 min.

5. The method for simultaneously extracting Rhizoma Drynariae starch and flavonoids according to claim 1, characterized in that: The filtration described in S1 uses a 100-200 mesh sieve.

6. The method for simultaneously extracting Rhizoma Drynariae starch and flavonoids according to claim 1, characterized in that: The centrifugation speed in S2 is 5000 rpm and the time is 20 min.

7. The method for simultaneously extracting Rhizoma Drynariae starch and flavonoids according to claim 1, characterized in that: The drying temperature in S2 is 40-50°C.

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