Gleditsia sinensis saponin extraction method
By combining gradient pressurization treatment with sulfonated hydroxypropyl-β-cyclodextrin, the problems of solvent residue and polysaccharide residue in saponin extraction from saponins were solved, achieving efficient and low-residue saponin extraction.
Patent Information
- Application Number
- CN202511456624.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-10-13
- Publication Date
- 2026-01-16
AI Technical Summary
Existing saponin extraction technologies from soapberry trees suffer from problems such as solvent residue, high polysaccharide residue, limited supercritical CO2 dissolution capacity, and long extraction time under high temperature and high pressure, making it difficult to achieve efficient and low-residue saponin extraction.
Gradient pressurization combined with sulfonated hydroxypropyl-β-cyclodextrin was used to break down the cell wall through negative charge repulsion and steric hindrance. After adding sulfonated hydroxypropyl-β-cyclodextrin, the pH value was adjusted for separation, and high-pressure microfluidic equipment was used for extraction.
It improves saponin extraction efficiency, reduces solvent and polysaccharide residues, avoids local high-temperature damage under high temperature and high pressure, and achieves efficient and low-residue saponin extraction.
Smart Images
Figure CN121342901A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of natural product extraction technology, specifically to a method for extracting saponins from saponins. Background Technology
[0002] In the field of natural product extraction, saponins (including oleanolic acid-type triterpenoid saponins), as important bioactive components, are widely used in the pharmaceutical, daily chemical, and biopesticide industries. Traditional extraction processes mainly rely on organic solvent extraction and supercritical fluid extraction, both of which have significant drawbacks.
[0003] Organic solvent extraction typically uses a 60-80% ethanol aqueous solution, refluxed at 70-80℃ for 4-6 hours. Experiments show that this process is prone to solvent residue problems, and the residual polysaccharides are high, directly reducing the bioactivity of the product.
[0004] To improve purity, the industry is gradually turning to supercritical CO2 extraction technology. Although this method can reduce solvent residue, it faces two bottlenecks: first, supercritical CO2 has limited ability to dissolve polar saponins; second, high-pressure extraction takes a long time.
[0005] In recent years, auxiliary extraction technologies, such as enzymatic hydrolysis, have also had limitations. Enzymatic hydrolysis is difficult to scale up for continuous production because cellulase / pectinase is expensive and easily deactivated at high temperatures.
[0006] Therefore, existing technologies need further development. Summary of the Invention
[0007] To address the shortcomings of existing technologies and solve the aforementioned problems, a method for extracting saponins from saponins from saponins is proposed, and the following technical solution is provided: A method for extracting saponins from soap pods includes: crushing soap pod shells into soap pod powder and mixing it with deionized water to obtain soap pod slurry; adding sulfonated hydroxypropyl-β-cyclodextrin to the soap pod slurry after gradient pressurization treatment; adjusting the pH value to acidic; and then separating to obtain saponins.
[0008] Furthermore, the gradient pressurization process specifically involves: processing at a pressure of 30-50 MPa for 0.3-0.6 seconds, then pressurizing to 80-100 MPa; processing at a pressure of 80-100 MPa for 0.3-0.6 seconds, then pressurizing to 120-150 MPa, and processing at a pressure of 120-150 MPa for 0.3-0.6 seconds.
[0009] Furthermore, a high-pressure microjet device is used for gradient pressurization.
[0010] Furthermore, the temperature is controlled at 30-40℃ during the gradient boosting process.
[0011] Furthermore, the amount of sulfonated hydroxypropyl-β-cyclodextrin added is 7-9% of the mass of the soapberry powder.
[0012] Furthermore, sulfonated hydroxypropyl-β-cyclodextrin was added within 30 seconds after the gradient pressurization treatment.
[0013] Furthermore, sulfonated hydroxypropyl-β-cyclodextrin is dissolved in deionized water to form a sulfonated hydroxypropyl-β-cyclodextrin solution with a concentration of 0.14-0.15 g / ml, and the sulfonated hydroxypropyl-β-cyclodextrin solution is added to the soapberry pulp at a rate of 10-15 ml / s.
[0014] Furthermore, the pH was adjusted to 3-5 using citrate buffer.
[0015] Furthermore, after adjusting the pH to acidic, the saponin was obtained by centrifugation and chromatographic purification.
[0016] Beneficial effects: 1. This invention breaks down cell walls through gradient pressurization, avoiding thermal degradation. Compared with single-stage high-pressure crushing, it improves efficiency and avoids local high temperatures caused by sudden pressure increases.
[0017] 2. This invention incorporates sulfonated hydroxypropyl-β-cyclodextrin, which blocks oxidative condensation through a dual mechanism: on the one hand, it repels negatively charged polysaccharides by negative charge, and on the other hand, it forms a physical barrier on the surface of saponins by steric hindrance. Attached Figure Description
[0018] Figure 1 This is the infrared spectrum of saponin obtained in Example 1 of the present invention. Detailed Implementation
[0019] To enable those skilled in the art to better understand the technical solutions of the present invention, the technical solutions of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Based on the embodiments in this application, other similar embodiments obtained by those skilled in the art without creative effort should all fall within the scope of protection of this application.
[0020] According to embodiments of the present invention, a method for extracting saponins from soap pods is provided, comprising: crushing soap pod shells into soap pod powder and mixing it with deionized water to obtain a soap pod slurry; subjecting the soap pod slurry to gradient pressurization treatment and adding sulfonated hydroxypropyl-β-cyclodextrin; adjusting the pH value to acidic; and then separating to obtain saponins. The present invention uses gradient pressurization treatment to break down cell walls and avoid thermal degradation, improving efficiency compared to single-stage high-pressure crushing and avoiding localized high temperatures caused by sudden pressure increases. The addition of sulfonated hydroxypropyl-β-cyclodextrin blocks oxidative condensation through a dual mechanism: on the one hand, it repels negatively charged polysaccharides through negative charge repulsion; on the other hand, it forms a physical barrier on the saponin surface through steric hindrance.
[0021] Example 1 The soapberry shells were crushed and passed through a 40-mesh sieve. 100 parts by weight of the dry powder were mixed with deionized water to prepare a soapberry slurry. The soapberry slurry was injected into a high-pressure microfluidic device and subjected to gradient pressurization: 50 MPa for 0.5 seconds → 100 MPa for 0.5 seconds → 150 MPa for 0.5 seconds, with the temperature controlled at 35℃ during the gradient pressurization process. Within 30 seconds after the gradient pressurization was completed, an aqueous solution of sulfonated hydroxypropyl-β-cyclodextrin was injected. The concentration of sulfonated hydroxypropyl-β-cyclodextrin was 0.15 g / ml, the amount of sulfonated hydroxypropyl-β-cyclodextrin added was 8 parts by weight, and the addition rate was 12 ml / s. The pH was adjusted to 4 with 0.5 M citrate buffer, and after stirring for 10 minutes, the mixture was centrifuged to precipitate. After dissociation with anhydrous ethanol for 30 minutes, the filtrate was loaded onto an AB-8 macroporous resin column. Impurities were first washed with 3 BV of water, and then eluted with 5 BV of 70% ethanol. The eluent was collected and freeze-dried to obtain refined saponin. The refined saponin was subjected to infrared testing, and the test results are as follows: Figure 1 As shown, the impurity peaks are significantly reduced.
[0022] Example 2 The soapberry shells were crushed and passed through a 40-mesh sieve. 100 parts by weight of the dry powder were mixed with deionized water to prepare a soapberry slurry. The soapberry slurry was injected into a high-pressure microfluidic device and subjected to gradient pressurization: 30 MPa for 0.6 seconds → 80 MPa for 0.3 seconds → 120 MPa for 0.6 seconds, with the temperature controlled at 30℃ during the gradient pressurization process. Within 30 seconds after the gradient pressurization was completed, an aqueous solution of sulfonated hydroxypropyl-β-cyclodextrin was injected. The concentration of sulfonated hydroxypropyl-β-cyclodextrin was 0.14 g / ml, the amount of sulfonated hydroxypropyl-β-cyclodextrin added was 7 parts by weight, and the addition rate was 10 ml / s. The pH was adjusted to 3 with 0.5 M citrate buffer, and after stirring for 10 minutes, the mixture was centrifuged to precipitate. After dissociation with anhydrous ethanol for 30 minutes, the filtrate was loaded onto an AB-8 macroporous resin column. Impurities were first washed with 3 BV of water, and then eluted with 5 BV of 70% ethanol. The eluent was collected and freeze-dried to obtain refined saponin.
[0023] Example 3 The soapberry shells were crushed and passed through a 40-mesh sieve. 100 parts by weight of the dry powder were mixed with deionized water to prepare a soapberry slurry. The soapberry slurry was injected into a high-pressure microfluidic device and subjected to gradient pressurization: 50 MPa for 0.3 seconds → 100 MPa for 0.3 seconds → 150 MPa for 0.3 seconds, with the temperature controlled at 40℃ during the gradient pressurization process. Within 30 seconds after the gradient pressurization was completed, an aqueous solution of sulfonated hydroxypropyl-β-cyclodextrin was injected. The concentration of sulfonated hydroxypropyl-β-cyclodextrin was 0.15 g / ml, the amount of sulfonated hydroxypropyl-β-cyclodextrin added was 9 parts by weight, and the addition rate was 15 ml / s. The pH was adjusted to 5 with 0.5 M citrate buffer, and after stirring for 10 minutes, the mixture was centrifuged to precipitate. After dissociation with anhydrous ethanol for 30 minutes, the filtrate was loaded onto an AB-8 macroporous resin column. Impurities were first washed with 3 BV of water, and then eluted with 5 BV of 70% ethanol. The eluent was collected and freeze-dried to obtain refined saponin.
[0024] Comparative Example 1 The soapberry shells were crushed and passed through a 40-mesh sieve. 100 parts by weight of the dry powder were mixed with deionized water to prepare a soapberry slurry. The soapberry slurry was injected into a high-pressure microfluidic device, maintained at 150 MPa for 1.5 seconds, and the temperature was controlled at 35℃. Within 30 seconds after the pressurization was completed, an aqueous solution of sulfonated hydroxypropyl-β-cyclodextrin was injected. The concentration of sulfonated hydroxypropyl-β-cyclodextrin was 0.14-0.15 g / ml, the amount of sulfonated hydroxypropyl-β-cyclodextrin added was 8 parts by weight, and the addition rate was 12 ml / s. The pH was adjusted to 4 with 0.5 M citrate buffer, and after stirring for 10 minutes, the mixture was centrifuged to precipitate. After dissociation with anhydrous ethanol for 30 minutes, the filtrate was loaded onto an AB-8 macroporous resin column. Impurities were first washed with 3 BV of water, and then eluted with 5 BV of 70% ethanol. The eluent was collected and freeze-dried to obtain refined saponin.
[0025] Comparative Example 2 The soap pod shells were crushed and passed through a 40-mesh sieve. 100 parts by weight of the dry powder were mixed with deionized water to prepare soap pod slurry. The soap pod slurry was injected into a high-pressure micro-jet device and subjected to gradient pressure increase: 50MPa for 0.5 seconds → 100MPa for 0.5 seconds → 150MPa for 0.5 seconds. The temperature was controlled at 35℃ during the gradient pressure increase process. After the gradient pressurization was completed, an aqueous solution of sulfonated hydroxypropyl-β-cyclodextrin was injected for 45 seconds. The concentration of sulfonated hydroxypropyl-β-cyclodextrin was 0.14-0.15 g / ml, the amount of sulfonated hydroxypropyl-β-cyclodextrin added was 8 parts by weight, and the addition rate was 12 ml / s. The pH was adjusted to 4 with 0.5 M citrate buffer, and after stirring for 10 minutes, the mixture was centrifuged to precipitate. After dissociation with anhydrous ethanol for 30 minutes, the filtrate was loaded onto an AB-8 macroporous resin column. The column was first washed with 3 BV of water, and then eluted with 5 BV of 70% ethanol. The eluent was collected and freeze-dried to obtain refined saponin.
[0026] Comparative Example 3 The soapberry shells were crushed and passed through a 40-mesh sieve. 100 parts by weight of the dry powder were mixed with deionized water to prepare a soapberry slurry. The slurry was injected into a high-pressure microfluidic device and subjected to a gradient pressurization: 50 MPa for 0.5 seconds → 100 MPa for 0.5 seconds → 150 MPa for 0.5 seconds, with the temperature controlled at 35℃ during the gradient pressurization process. After the gradient pressurization was completed, 0.5 M citrate buffer was added to adjust the pH to 4. After stirring for 10 minutes, the mixture was centrifuged to precipitate. The precipitate was then dissociated with anhydrous ethanol for 30 minutes, and the filtrate was loaded onto an AB-8 macroporous resin column. The column was first washed with 3 BV of water, then eluted with 5 BV of 70% ethanol. The eluent was collected and freeze-dried to obtain refined saponin.
[0027] The saponin extraction efficiency of Examples 1-3 and Comparative Examples 1-3 was compared, and the comparison results are shown in Table 1.
[0028] Saponin purity was determined by HPLC, and polysaccharide residues were determined by the phenol-sulfuric acid method.
[0029] Table 1. Comparison of saponin extraction efficiency in Examples 1-3 and Comparative Examples 1-3 As shown in Table 1 above, the saponin extraction efficiency of this embodiment is higher than that of the comparative example. Specifically, when the gradient pressurization method is not used, the saponin yield decreases, and the cell wall cannot be completely destroyed. In addition, the absence of sulfonated hydroxypropyl-β-cyclodextrin or the late addition of sulfonated hydroxypropyl-β-cyclodextrin also affects the saponin yield. This application has developed a novel saponin extraction method by combining a gradient pressurization method with sulfonated hydroxypropyl-β-cyclodextrin.
[0030] Furthermore, it should be understood that although this specification describes embodiments, not every embodiment contains only one independent technical solution. This narrative style is merely for clarity. Those skilled in the art should consider the specification as a whole, and the technical solutions in each embodiment can also be appropriately combined to form other embodiments that can be understood by those skilled in the art.
Claims
1. A method for extracting saponin from Gleditsia sinensis L., characterized in that, The method comprises the following steps: The Gleditsia sinensis shell is crushed into Gleditsia sinensis powder, which is mixed with deionized water to obtain a Gleditsia sinensis slurry. The Gleditsia sinensis slurry is treated by gradient pressure increase, and then sulfonated hydroxypropyl-β-cyclodextrin is added. After the pH value is adjusted to be acidic, Gleditsia sinensis is obtained by separation.
2. The method of extracting saponin from Gleditsia sinensis according to claim 1, characterized in that, The gradient pressure increase treatment is specifically as follows: the pressure is treated at 30-50 MPa for 0.3-0.6 seconds, and then the pressure is increased to 80-100 MPa; the pressure is treated at 80-100 MPa for 0.3-0.6 seconds, and then the pressure is increased to 120-150 MPa, and the pressure is treated at 120-150 MPa for 0.3-0.6 seconds.
3. The method of extracting saponin from Gleditsia sinensis according to claim 1, characterized in that, The gradient pressure increase treatment is performed by using a high-pressure microfluidic device.
4. The method of extracting saponin from Gleditsia sinensis according to claim 1, characterized in that, The temperature is controlled to be 30-40 ℃ during the gradient pressure increase treatment.
5. The method of extracting saponin from Gleditsia sinensis according to claim 1, characterized in that, The sulfonated hydroxypropyl-β-cyclodextrin is added in an amount of 7-9% of the mass of the Gleditsia sinensis powder.
6. The method of extracting Gleditsin according to claim 1, wherein, The sulfonated hydroxypropyl-β-cyclodextrin is added within 30 seconds after the gradient pressure increase treatment.
7. The method of extracting saponin from Gleditsia sinensis according to claim 1, characterized in that, The sulfonated hydroxypropyl-β-cyclodextrin is dissolved in deionized water to form a sulfonated hydroxypropyl-β-cyclodextrin solution with a concentration of 0.14-0.15 g / ml, and the sulfonated hydroxypropyl-β-cyclodextrin solution is added to the Gleditsia sinensis slurry at a speed of 10-15 ml / s.
8. The method of extracting saponin from Gleditsia sinensis according to claim 1, characterized in that, The pH value is adjusted to be 3-5 by using a citric acid buffer.
9. The method of extracting saponin from Gleditsia sinensis according to claim 1, characterized in that, After the pH value is adjusted to be acidic, Gleditsia sinensis is obtained by centrifugal separation and chromatographic purification.