Radix rehmanniae preparata high active ingredient preservation extraction and stock solution preparation method

Abstract

The application discloses a kind of high active component preservation extraction and stock solution preparation method of Radix Rehmanniae Preparata, belongs to the technical field of traditional Chinese medicine deep processing and liquid preparation preparation, and specifically relates to a kind of high active component preservation extraction and stock solution preparation method of Radix Rehmanniae Preparata, for solving the problems of loss of heat-sensitive components in traditional high-temperature decoction and insufficient stability of stock solution.The method comprises the following steps: taking Radix Rehmanniae Preparata raw materials, drying at low temperature and then crushing;mixing the Radix Rehmanniae Preparata powder with purified water, and low-temperature segment extraction under the condition that the temperature does not exceed 60 DEG C in an increasing temperature mode;after the extraction solution is combined, it is subjected to coarse filtration, centrifugation, fine filtration, low-temperature vacuum concentration, sterile filtration and sterile filling to obtain the Radix Rehmanniae Preparata stock solution.In the examples, the retention rate of gamma-aminobutyric acid in the obtained stock solution is 78%, the dissolution rate of radix rehmanniae polysaccharide is 86%, and the light transmittance is 96%, which is conducive to the consideration of water-soluble component dissolution, heat-sensitive component preservation and stock solution clarity and stability.

Description

Technical Field

[0001] This invention belongs to the field of deep processing of traditional Chinese medicine and preparation of liquid preparations, specifically relating to a method for preserving and extracting highly active ingredients from Rehmannia glutinosa and preparing the original solution. Background Technology

[0002] Prepared Rehmannia glutinosa is a commonly used Chinese medicinal material or processed medicinal slices obtained by steaming, sun-drying, or drying Rehmannia glutinosa. It is often used in decoctions, pastes, pills, granules, and related liquid products that are both food and medicine. With the development of deep-processed Rehmannia glutinosa products, liquid products such as prepared Rehmannia glutinosa extract, concentrated liquid, and oral liquid are gradually gaining attention.

[0003] The preparation of liquid Rehmannia glutinosa products currently involves methods such as high-temperature decoction, prolonged reflux extraction, or direct concentrated decoction. While these processes can extract some water-soluble components, they still have the following drawbacks: Firstly, the dissolution conditions of components such as Rehmannia glutinosa polysaccharides, glycosides, amino acids, and small molecule alcohols are not entirely the same, and high-temperature decoction alone can easily lead to uneven release of components. Secondly, heat-sensitive or processing-sensitive components such as γ-aminobutyric acid, free amino acids, and natural glycerol are prone to loss or degradation during prolonged high-temperature processing, resulting in a decrease in the integrity of the components in the final product. In addition, traditional decoctions often contain a large amount of insoluble fine residue, large molecular redundant substances, and colloidal impurities, affecting the clarity and storage stability of the liquid product and increasing the difficulty of subsequent filling and processing.

[0004] Therefore, it is necessary to provide a low-temperature extraction and stock solution preparation method suitable for the preparation of liquid Rehmannia glutinosa products, which can ensure the full dissolution of effective water-soluble components, minimize the loss of heat-sensitive active ingredients, and improve the purity and processing stability of the stock solution. Summary of the Invention

[0005] In view of the above situation and to overcome the defects of the prior art, the present invention provides a method for preserving and extracting high-activity components of Rehmannia glutinosa and preparing the stock solution. The method uses Rehmannia glutinosa raw material as the extraction target, and obtains Rehmannia glutinosa stock solution by low-temperature segmented extraction with increasing temperature, multi-stage impurity removal, low-temperature vacuum concentration and aseptic fine filtration and filling.

[0006] This invention proposes a method for preserving and extracting highly active components from Rehmannia glutinosa and preparing the stock solution, comprising the following steps: S1. Raw material pretreatment: Take prepared Rehmannia glutinosa raw material, dry it at low temperature and then pulverize it to obtain prepared Rehmannia glutinosa powder; S2. Low-temperature fractional extraction: The prepared Rehmannia glutinosa powder is mixed with purified water, and at least two stages of water extraction are carried out at a temperature not exceeding 60°C in an incremental manner to obtain each stage of extract. S3. Merging and removing impurities: Combine the extracts from each segment and then perform coarse filtration, centrifugation and fine filtration in sequence to remove insoluble impurities and obtain a clear extract; S4. Low-temperature vacuum concentration: The clarified extract is concentrated at low temperature under reduced pressure to obtain Rehmannia glutinosa concentrate; S5. Aseptic processing and filling: The prepared Rehmannia glutinosa concentrate is aseptically filtered and filled under aseptic conditions to obtain prepared Rehmannia glutinosa stock solution.

[0007] Furthermore, the prepared Rehmannia glutinosa raw material mentioned in step S1 is prepared Rehmannia glutinosa that has undergone nine steaming and nine sun-drying processes, preferably prepared Rehmannia glutinosa obtained through soaking to aid penetration, repeated steaming, sun-drying or low-temperature drying, and aging. Using this type of prepared Rehmannia glutinosa as raw material is beneficial for obtaining extraction raw materials with sufficient tissue softening and better conditions for component release.

[0008] Furthermore, the low-temperature drying in step S1 is carried out at a temperature of 35-55°C until the moisture content of the prepared Rehmannia glutinosa is 8%-18%. This drying condition helps to reduce the difficulty of pulverization and avoids the adverse effects of high-temperature drying on heat-sensitive components.

[0009] Furthermore, the particle size of the prepared Rehmannia glutinosa powder mentioned in step S1 is 80-300 mesh. Pulverization increases the contact area between the raw material and the extraction solvent, which helps to shorten the low-temperature extraction time and promote the dissolution of components.

[0010] Further, the mass-to-volume ratio of Rehmannia glutinosa powder to purified water in step S2 is from 1 kg:6 L to 1 kg:15 L.

[0011] Furthermore, the low-temperature segmented extraction described in step S2 includes: a first extraction at 25-35℃ for 0.5-2 hours; a second extraction at 38-48℃ for 1-3 hours; and a third extraction at 50-60℃ for 0.5-2 hours. This segmented extraction method balances low-temperature protection with gradual dissolution of components, reducing component loss caused by prolonged high-temperature cooking in a single step.

[0012] Furthermore, in step S3, the centrifugation speed is 3000-8000 r / min, and the centrifugation time is 5-20 minutes; the fine filtration is performed using a filter membrane or filter element with a pore size of 0.45-1.0 μm.

[0013] Furthermore, the concentration temperature of the low-temperature vacuum concentration in step S4 is 35-55℃, and the vacuum degree is -0.06MPa to -0.095MPa.

[0014] Furthermore, the soluble solids content of the Rehmannia glutinosa concentrate in step S4 is 10-35°Brix.

[0015] Furthermore, the aseptic filtration in step S5 is performed using a filter membrane or filter element with a pore size of 0.22 μm, and the prepared Rehmannia glutinosa extract obtained after filling is prepared Rehmannia glutinosa extract without added sucrose, honey, thickeners or preservatives.

[0016] By adopting the above solution, the present invention has the following beneficial effects: (1) The present invention uses Rehmannia glutinosa raw material and performs low-temperature drying and pulverization before extraction, which is beneficial to improving the extractability of raw material tissue and the uniformity of component release; when the Rehmannia glutinosa raw material is Rehmannia glutinosa processed by nine steaming and nine sun-drying, the characteristics of tissue softening and better component release conditions after processing can be further utilized.

[0017] (2) The present invention adopts a low-temperature segmented extraction method with a temperature not higher than 60°C and an increasing temperature, which is beneficial to take into account the dissolution of water-soluble components such as Rehmannia polysaccharides and glycosides, as well as the preservation of components that are easily affected by high temperature, such as γ-aminobutyric acid, free amino acids, and natural glycerol, and reduces the adverse effects of traditional high-temperature long-term decoction on the component system.

[0018] (3) The present invention removes impurities through coarse filtration, centrifugation and fine filtration in multiple stages, which can reduce the content of insoluble fine residue, colloidal impurities and redundant macromolecular substances in the extract, thereby improving the clarity of the original solution and its suitability for subsequent filling.

[0019] (4) The present invention uses low temperature vacuum concentration and aseptic fine filtration filling, which is beneficial to obtain Rehmannia glutinosa raw liquid with a certain soluble solids content under a lower heat load and improve the product processing stability. The small molecule alcohol components such as natural glycerol contained in Rehmannia glutinosa raw material are beneficial to improve the solubility of some water-soluble components and the stability of the system. Combined with multi-stage impurity removal process, the clarity of the raw liquid and the stability of the system can be further improved. Attached Figure Description

[0020] Figure 1 This is a process flow diagram of the method for preserving and extracting highly active components of Rehmannia glutinosa and preparing the original solution according to the present invention.

[0021] The accompanying drawings are provided to further illustrate the invention and form part of the specification. They are used together with the embodiments of the invention to explain the invention and do not constitute a limitation thereof. Detailed Implementation

[0022] The technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings of the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention. Example

[0023] A method for preserving and extracting highly active components from Rehmannia glutinosa and preparing its stock solution includes the following steps: S1. Raw material pretreatment: Take 10 kg of Rehmannia glutinosa that has been steamed and dried nine times and aged, dry it at a low temperature of 45℃ until the moisture content is 12%, pulverize it and pass it through a 120-mesh sieve to obtain Rehmannia glutinosa powder.

[0024] S2. Low-temperature fractional extraction: Add Rehmannia glutinosa powder to 90L of purified water, stir to disperse, and then perform three-stage extraction; the first stage is extracted at 30℃ for 1 hour, the second stage is extracted at 42℃ for 2 hours, and the third stage is extracted at 55℃ for 1 hour. Keep stirring slowly during the extraction process, and collect the extract from each stage separately.

[0025] S3. Merging and removing impurities: Combine the extracts from each segment, coarsely filter them through a 60-mesh sieve, centrifuge at 5000 r / min for 10 minutes, take the supernatant, and then finely filter it through a 0.65 μm filter cartridge to obtain a clear extract.

[0026] S4. Low-temperature vacuum concentration: The clarified extract was concentrated under reduced pressure at 45℃ and -0.085MPa until the soluble solids content was 20°Brix, to obtain the Rehmannia glutinosa concentrate.

[0027] S5. Aseptic processing and filling: The concentrated Rehmannia glutinosa extract is aseptically filtered through a 0.22μm filter membrane and filled under aseptic conditions to obtain the original Rehmannia glutinosa extract.

[0028] The prepared Rehmannia glutinosa extract obtained in this embodiment is brownish-brown in color, homogeneous in system, and contains no added sucrose, honey, thickeners or preservatives. It is suitable for use as a raw material for prepared Rehmannia glutinosa liquid products or oral liquid. Example

[0029] A method for preserving and extracting highly active components from Rehmannia glutinosa and preparing its stock solution includes the following steps: S1. Take 8 kg of Rehmannia glutinosa that has been processed by nine steaming and nine sun drying, dry it at a low temperature of 40℃ until the moisture content is 15%, pulverize it and pass it through an 80-mesh sieve to obtain Rehmannia glutinosa powder.

[0030] S2. Mix Rehmannia glutinosa powder with 70L of purified water and perform two-stage low-temperature extraction; the first stage is extracted at 32℃ for 1.5 hours, and the second stage is extracted at 50℃ for 2 hours. Collect the extract.

[0031] S3. First, the extract is coarsely filtered to remove larger particles, then centrifuged at 4000 r / min for 15 minutes. The supernatant is collected and filtered through a 1.0 μm filter to obtain a clear extract.

[0032] S4. Concentrate the clarified extract under low temperature vacuum at 50℃ and -0.075MPa until the soluble solids content is 15°Brix.

[0033] S5. The solution was aseptically filtered through a 0.22μm filter membrane and aseptically filled to obtain the prepared Rehmannia glutinosa extract.

[0034] This embodiment uses a two-stage low-temperature extraction method, which is suitable for production scenarios where the extraction time requirement is relatively short. Example

[0035] A method for preserving and extracting highly active components from Rehmannia glutinosa and preparing its stock solution includes the following steps: S1. Take 12 kg of Rehmannia glutinosa that has been soaked and osmotically enhanced, steamed nine times, sun-dried nine times, or dried at low temperature and aged. Dry it at 50℃ until the moisture content is 10%, pulverize it and pass it through a 200-mesh sieve to obtain Rehmannia glutinosa powder.

[0036] S2. Mix Rehmannia glutinosa powder with 150L of purified water. Extract the first stage at 28℃ for 1 hour, the second stage at 45℃ for 2.5 hours, and the third stage at 58℃ for 1 hour.

[0037] S3. Combine the extracts, filter coarsely, centrifuge at 7000 r / min for 8 minutes, collect the supernatant, and then filter finely through a 0.45 μm filter membrane to obtain a clear extract.

[0038] S4. The clarified extract was concentrated under reduced pressure at 42℃ and -0.09MPa until the soluble solids content was 30°Brix, to obtain the Rehmannia glutinosa concentrate.

[0039] S5. The concentrated Rehmannia glutinosa extract is aseptically filtered through a 0.22μm filter cartridge and aseptically filled to obtain the original Rehmannia glutinosa extract.

[0040] This embodiment combines finer grinding with three-stage extraction, which helps to improve the adequacy of extraction under low-temperature conditions.

[0041] Comparative Example 1 Rehmannia glutinosa extract was prepared using a traditional high-temperature decoction method. 10 kg of the same Rehmannia glutinosa raw material as in Example 1 was taken, 90 L of purified water was added, and the mixture was decocted at 95-100 °C for 3 hours. After coarse filtration, the extract was directly concentrated to 20 ° Brix.

[0042] The resulting concentrated solution was used as a control sample for high-temperature decoction.

[0043] Comparative Example 2 Take 10 kg of the same prepared Rehmannia glutinosa raw material as in Example 1, crush it, and extract it at 30°C for 4 hours. After coarse filtration, centrifugation and fine filtration, concentrate it to a soluble solids content of 14°Brix.

[0044] The resulting concentrate was used as a control sample for room temperature extraction.

[0045] Experimental Example 1 The samples obtained in Example 1, Comparative Example 1, and Comparative Example 2 were compared and quantitatively detected. The quantitative detection methods for the main active ingredients in Rehmannia glutinosa extract are as follows: γ-aminobutyric acid was determined by pre-column derivatization high-performance liquid chromatography (HPLC-FLD) using o-phthalaldehyde (OPA); catalpol and verbascoside were determined by high-performance liquid chromatography (HPLC-UV); free amino acids were determined by an automated amino acid analyzer; Rehmannia glutinosa polysaccharides were determined by the phenol-sulfuric acid method (calculated as glucose); soluble solids content was determined by a handheld refractometer; transmittance was determined by a UV-Vis spectrophotometer at 660 nm, with purified water as a blank.

[0046] Take 10g of Rehmannia glutinosa sample that has undergone nine steaming and nine sun-drying processes and aging, pulverize it and pass it through an 80-mesh sieve. Prepare test solutions according to the water extraction method in Chapter 2201 of the General Section IV of the 2020 edition of the Chinese Pharmacopoeia. Take samples to determine the content of the above-mentioned components, and use the results as the raw material reference values. For each sample of Example 1, Comparative Example 1 and Comparative Example 2, take samples for testing under the same feed amount (based on the amount of raw drug), and calculate the dissolution rate and retention rate of each component. The results are shown in Table 1.

[0047] Table 1. Calculated content and retention / dissolution rate of each component in samples obtained by different extraction methods. γ-Aminobutyric acid (GABA) 3.2 mg / g 2.5 mg / g (78% retention) 1.3 mg / g (41% retention) 2.5 mg / g (78% retention) Total amount of free amino acids 17.0 mg / g 13.8 mg / g (81% retention) 7.5 mg / g (44% retention) 14.5 mg / g (85% retention) Zizhu alcohol 6.0 mg / g 4.8 mg / g (dissolution rate 80%) 5.5 mg / g (dissolution rate 92%) 3.8 mg / g (dissolution rate 63%) Verbascoside 1.9 mg / g 1.5 mg / g (79% retention) 0.7 mg / g (retention rate 37%) 1.6 mg / g (84% retention) Rehmannia polysaccharide (calculated as glucose) 94.0 mg / g 81.0 mg / g (dissolution rate 86%) 64.0 mg / g (dissolution rate 68%) 58.0 mg / g (dissolution rate 62%) Soluble solids content not applicable 20°Brix 20°Brix 14°Brix Note: All data are averages of three independent tests; the raw material baseline values ​​are based on the dry weight of the raw medicinal materials, and the data for each sample are converted to the corresponding content per gram of dry weight of raw medicinal materials based on the same amount of feed. Among them, γ-aminobutyric acid, total free amino acids and verbascoside are evaluated by retention rate, while catalpol and Rehmannia polysaccharide are evaluated by dissolution rate.

[0048] As shown in Table 1: (1) Preservation effect of heat-sensitive components: The γ-aminobutyric acid retention rate of Example 1 was 78%, which was higher than 41% of Comparative Example 1 (an increase of 37 percentage points); the total free amino acid retention rate was 81%, which was higher than 44% of Comparative Example 1 (an increase of 37 percentage points); and the verbascoside retention rate was 79%, which was higher than 37% of Comparative Example 1 (an increase of 42 percentage points). The above results indicate that low-temperature fractional extraction can reduce the damage of heat-sensitive components caused by high temperature.

[0049] (2) Solubility of polysaccharide components: The dissolution rate of Rehmannia glutinosa polysaccharide in Example 1 was 86%, which was higher than that of Comparative Example 2 (62%, an increase of 24 percentage points), indicating that segmented heating extraction can more fully dissolve polysaccharide macromolecular components compared with single room temperature long-term extraction.

[0050] (3) Overall performance of glycoside components: In Example 1, the dissolution rate of catalpol was 80%, which was slightly lower than that of Comparative Example 1 (92%), but it balanced the preservation of heat-sensitive components and the sufficiency of dissolution. The overall integrity of the active ingredients was better than that of Comparative Example 1.

[0051] Example 1 is close to the room temperature extraction control in terms of preservation of heat-sensitive components, and is superior to the room temperature extraction control in terms of sufficient dissolution of water-soluble components and clarification stability. At the same time, compared with the high temperature decoction control, the stock solution obtained in Example 1 has a more uniform appearance, higher light transmittance, and less precipitation after standing.

[0052] This invention employs low-temperature segmented extraction (30℃→42℃→55℃) instead of traditional high-temperature long-term decoction (95-100℃, 3 hours), which increases the retention rate of heat-sensitive components such as γ-aminobutyric acid from 41% to 78%, while simultaneously increasing the dissolution rate of Rehmannia glutinosa polysaccharides from 62% to 86%. This effectively resolves the contradiction between "preserving heat-sensitive components" and "fully dissolving polysaccharides" in traditional processes. Furthermore, through multi-stage impurity removal (coarse filtration, centrifugation, and fine filtration) and low-temperature vacuum concentration, the resulting stock solution has higher transmittance, less precipitation after settling, and better system stability.

[0053] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A method for preserving and extracting highly active components from Rehmannia glutinosa and preparing its stock solution, characterized in that, Includes the following steps: S1. Raw material pretreatment: Take prepared Rehmannia glutinosa raw material, dry it at low temperature and then pulverize it to obtain prepared Rehmannia glutinosa powder; S2. Low-temperature fractional extraction: The prepared Rehmannia glutinosa powder is mixed with purified water, and at least two stages of water extraction are carried out at a temperature not exceeding 60°C in an incremental manner to obtain each stage of extract. S3. Merging and removing impurities: Combine the extracts from each segment and then perform coarse filtration, centrifugation and fine filtration in sequence to remove insoluble impurities and obtain a clear extract; S4. Low-temperature vacuum concentration: The clarified extract is concentrated at low temperature under reduced pressure to obtain Rehmannia glutinosa concentrate; S5. Aseptic processing and filling: The prepared Rehmannia glutinosa concentrate is aseptically filtered and filled under aseptic conditions to obtain prepared Rehmannia glutinosa stock solution.

2. The method for preserving and extracting highly active components of Rehmannia glutinosa and preparing the stock solution according to claim 1, characterized in that, The prepared Rehmannia glutinosa raw material mentioned in step S1 is prepared Rehmannia glutinosa that has been processed by nine steaming and nine sun-drying processes. The prepared Rehmannia glutinosa that has been processed by nine steaming and nine sun-drying processes is prepared Rehmannia glutinosa that has been soaked to aid penetration, repeatedly steamed, sun-dried or dried at low temperature and aged.

3. The method for preserving and extracting highly active components of Rehmannia glutinosa and preparing the stock solution according to claim 1, characterized in that, The low-temperature drying in step S1 is carried out at a temperature of 35-55℃ until the moisture content of the prepared Rehmannia glutinosa is 8%-18%.

4. The method for preserving and extracting highly active components of Rehmannia glutinosa and preparing the stock solution according to claim 1, characterized in that, The particle size of the prepared Rehmannia glutinosa powder mentioned in step S1 is 80-300 mesh.

5. The method for preserving and extracting highly active components of Rehmannia glutinosa and preparing the stock solution according to claim 1, characterized in that, The mass-to-volume ratio of Rehmannia glutinosa powder to purified water in step S2 is from 1 kg:6 L to 1 kg:15 L.

6. The method for preserving and extracting highly active components of Rehmannia glutinosa and preparing the stock solution according to claim 1, characterized in that, The low-temperature fractional extraction in step S2 includes: The first stage of extraction involves extraction at 25-35℃ for 0.5-2 hours; The second stage of extraction involves extraction at 38-48℃ for 1-3 hours; The third stage of extraction involves extraction at 50-60℃ for 0.5-2 hours.

7. The method for preserving and extracting highly active components of Rehmannia glutinosa and preparing the stock solution according to claim 1, characterized in that, In step S3, the centrifugation speed is 3000-8000 r / min and the centrifugation time is 5-20 minutes; the fine filtration is carried out using a filter membrane or filter element with a pore size of 0.45-1.0 μm.

8. The method for preserving and extracting highly active components of Rehmannia glutinosa and preparing the stock solution according to claim 1, characterized in that, The concentration temperature of the low-temperature vacuum concentration in step S4 is 35-55℃, and the vacuum degree is -0.06MPa to -0.095MPa.

9. The method for preserving and extracting highly active components of Rehmannia glutinosa and preparing the stock solution according to claim 1, characterized in that, The soluble solids content of the prepared Rehmannia glutinosa concentrate in step S4 is 10-35°Brix; the aseptic fine filtration in step S5 is carried out using a filter membrane or filter element with a pore size of 0.22μm.

10. A prepared Rehmannia glutinosa extract, characterized in that, The prepared Rehmannia glutinosa extract is obtained by the method for preserving and extracting the highly active components of Rehmannia glutinosa according to any one of claims 1-9, wherein the Rehmannia glutinosa extract does not contain sucrose, honey, thickeners or preservatives.

Images