Method for removing alkaloids from traditional chinese medicine extracts

Abstract

The application discloses a method for removing alkaloids from traditional Chinese medicine extracts, wherein the traditional Chinese medicine extracts include Senecio scandens extracts, the alkaloids include adonifoline (PAs), and the method comprises the following steps: (1) water extraction of Senecio scandens under acidic conditions to obtain a first extraction liquid; (2) reaction of the obtained first extraction liquid under catalysis of inorganic acid to obtain a second extraction liquid; and (3) plate-frame filtration and ceramic membrane filtration of the obtained second extraction liquid, and drying of the filtrate to obtain the Senecio scandens extracts. The method for removing adonifoline (PAs) from the Senecio scandens extracts can greatly reduce the content of adonifoline (PAs) in the extracts, and has the advantages of simple process, low cost, non-toxicity and harmlessness, and simplicity and reliability.

Description

Technical Field

[0001] This invention relates to the field of extraction and separation technology of effective components of traditional Chinese medicine, specifically to a method for removing alkaloids from traditional Chinese medicine extracts, wherein the traditional Chinese medicine extract includes Senecio scandens extract, and the alkaloids include adoniflurine (PAs). Background Technology

[0002] Senecio scandens is the dried aerial part of the plant Senecio scandens, belonging to the Asteraceae family. Senecio scandens has a long history of medicinal use; it was first recorded in the Tang Dynasty's *Compendium of Materia Medica* and later included in the *Chinese Pharmacopoeia*. Its effects include clearing heat and detoxifying, promoting diuresis, and improving eyesight. Therefore, it is often used clinically to treat skin eczema, red and swollen eyes, and damp-heat dysentery, and is particularly effective in treating urticaria.

[0003] Senecio scandens contains various active ingredients, mainly including volatile oils, flavonoids, inorganic acids, and pyrrolizidine alkaloids (PAs). Pyrrolizidine alkaloids (PAs) have significant hepatotoxicity, and adoniflunline, as a characteristic PA in Senecio scandens, is subject to corresponding limits. The Chinese Pharmacopoeia stipulates that the content of adoniflunline in Senecio scandens must not exceed 0.004%. Based on this, the present invention aims to provide a method for removing adoniflunline from Senecio scandens extract. Summary of the Invention

[0004] Purpose of the invention: The technical problem to be solved by the present invention is to provide a method for preparing Senecio scandens extract with adoniflunline removed, which addresses the shortcomings of the prior art.

[0005] Invention concept: This invention adds acidic conditions to the extraction process of Senecio scandens, followed by inorganic acid catalytic precipitation, plate and frame filtration, and ceramic membrane clarification to effectively reduce the adonicoline content. Then, inorganic nanofiltration membrane is used to separate the effective components, followed by drying, to finally obtain a Senecio scandens extract that is basically free of adonicoline.

[0006] To solve the above-mentioned technical problems, the present invention discloses the following technical solution:

[0007] A method for preparing Senecio scandens extract includes the following steps:

[0008] (1) The first extract was obtained by water extraction of Senecio scandens under acidic conditions;

[0009] (2) The first extract obtained is reacted under the catalysis of inorganic acid to obtain the second extract;

[0010] (3) The second extract obtained is filtered by plate and frame filtration and ceramic membrane filtration. The filtrate is dried to obtain the Senecio scandens extract. Adoniflunine in the obtained Senecio scandens extract is almost completely removed.

[0011] In step (1), the pH of the acidic conditions is 2-5; preferably, the acidic conditions are any one or more combinations of hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid and citric acid.

[0012] In step (1), the extraction temperature is 60-100℃.

[0013] In step (2), the inorganic acid is any one or a combination of formaldehyde sulfuric acid, ammonium molybdate sulfuric acid, silicotungstic acid, and phosphomolybdic acid; preferably, the formaldehyde sulfuric acid is a composition of formaldehyde and sulfuric acid, with a volume ratio of formaldehyde to sulfuric acid of 1:5-7, more preferably 1:6; preferably, the ammonium molybdate sulfuric acid is a composition of ammonium molybdate and sulfuric acid, with a volume ratio of ammonium molybdate to sulfuric acid of 1:95-103, more preferably 1:99.

[0014] In step (2), the amount of inorganic acid used is 0.1%-1% of the volume or mass of the first extract.

[0015] In step (2), the reaction temperature is 20-70℃, preferably 30-60℃.

[0016] In step (2), the reaction time is 3-20 min, preferably 5-15 min.

[0017] In step (3), the aperture of the plate frame is 0.5-15μm, preferably 1-10μm, and more preferably 2-10μm.

[0018] In step (3), the pore size of the ceramic membrane is 5-50 nm; preferably, the pressure of the ceramic membrane filtration is 0.1-0.5 MPa; preferably, the flow rate of the ceramic membrane filtration is 1-4 m / s; preferably, the temperature of the ceramic membrane filtration is 20-50 °C.

[0019] The Senecio scandens extract prepared by the above method is also within the scope of protection of this invention, wherein the adoniflurine content in the Senecio scandens extract is less than 0.001%.

[0020] Beneficial effects:

[0021] Currently, with the improvement of medicinal material quality and extraction technology, the utilization rate of *Senecio scandens* is gradually increasing. However, this is accompanied by an increase in the content of adoniflurine (PAs). To address this, various conventional separation methods, such as macroporous resin adsorption, inorganic solvent extraction, and silica gel column chromatography, suffer from significant problems including high investment, high cost, and complex processes. Therefore, there is an urgent need to find a low-cost, simple method to remove or reduce the content of adoniflurine. The method for removing adoniflurine from *Senecio scandens* extract provided by this invention can significantly reduce the content of adoniflurine, and the process is simple, low-cost, non-toxic, harmless, and reliable. Attached Figure Description

[0022] The present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments, and the advantages of the present invention in the above and / or other aspects will become clearer.

[0023] Figure 1 These are the HPLC-MS parameters for adoniflunline.

[0024] Figure 2 This is one of the HPLC-MS chromatograms of adoniflurine (ADNFLJ) standards at different concentrations determined by the internal standard method of wild lily alkaloid (YBHJ).

[0025] Figure 3 This is one of the HPLC-MS chromatograms of adoniflurine (ADNFLJ) standards at different concentrations determined by the internal standard method of wild lily alkaloid (YBHJ).

[0026] Figure 4 This is one of the HPLC-MS chromatograms of adoniflurine (ADNFLJ) standards at different concentrations determined by the internal standard method of wild lily alkaloid (YBHJ).

[0027] Figure 5 This is one of the HPLC-MS chromatograms of adoniflurine (ADNFLJ) standards at different concentrations determined by the internal standard method of wild lily alkaloid (YBHJ).

[0028] Figure 6 The image shows the HPLC-MS chromatogram of adoniflurine (PAs) in the original extract of Senecio scandens.

[0029] Figure 7 The image shows the HPLC chromatogram of chlorogenic acid in the original extract of Senecio scandens.

[0030] Figure 8 The image shows the HPLC chromatogram of hyperoside in the original extract of Senecio scandens.

[0031] Figure 9 This is an HPLC-MS chromatogram of adoniflurine (PAs) in the clarified liquid after clarification using a ceramic membrane in a conventional process.

[0032] Figure 10 This is an HPLC chromatogram of chlorogenic acid in the clarified liquid after a conventional ceramic membrane process.

[0033] Figure 11 This is the HPLC chromatogram of hyperoside in the clarified liquid after clarification using a ceramic membrane in a conventional process.

[0034] Figure 12 This is an HPLC-MS chromatogram of adoniflurine (PAs) in the concentrate after clarification using a ceramic membrane following a conventional process.

[0035] Figure 13 This is an HPLC chromatogram of chlorogenic acid in the concentrated solution after clarification using a ceramic membrane following a conventional process.

[0036] Figure 14 This is the HPLC chromatogram of hyperoside in the concentrated solution after clarification using a ceramic membrane following a conventional process.

[0037] Figure 15 To improve the process, HPLC-MS chromatogram of adoniflurine (PAs) in the clarified liquid after clarification of the ceramic membrane was obtained.

[0038] Figure 16 To improve the process, HPLC chromatogram of chlorogenic acid in the clarified liquid after the ceramic membrane was clarified was obtained.

[0039] Figure 17 To improve the process, the HPLC chromatogram of hyperoside in the clarified liquid after the ceramic membrane was clarified was obtained.

[0040] Figure 18 To improve the process, HPLC-MS chromatograms of adoniflurine (PAs) in the concentrated solution after clarification using a ceramic membrane were obtained.

[0041] Figure 19 To improve the process, HPLC chromatogram of chlorogenic acid in the concentrated solution after clarification by ceramic membrane was obtained.

[0042] Figure 20 To improve the process, HPLC chromatogram of hyperoside in the concentrated solution after clarification using a ceramic membrane was obtained. Detailed Implementation

[0043] Unless otherwise specified, the experimental methods described in the following examples are conventional methods; unless otherwise specified, the reagents and materials are commercially available.

[0044] In the following examples, the method for removing adoniflunline from Senecio scandens extract includes (1) pulverizing Senecio scandens herb, adding a certain proportion of hot water, adjusting the pH and extracting twice, and combining the extracts; (2) adding a certain proportion of catalyst to the extract and catalyzing the reaction under certain temperature and time conditions; (3) clarifying the extract with a ceramic microfiltration membrane to obtain Senecio scandens ceramic membrane clarified liquid, which is then dried in an oven at 80°C to obtain Senecio scandens extract that is basically free of adoniflunline (PAs).

[0045] Example 1: Improved process for Senecio scandens

[0046] (1) Take a certain amount of Senecio scandens, add water at 60℃ and decoct twice, each time for 2 hours, add 10 times the amount of water each time, and add hydrochloric acid to adjust the pH to 2. Extract twice, combine the decoctions and set aside.

[0047] (2) Add 0.1% (v / v) of formaldehyde sulfuric acid to the extract of Senecio scandens for catalysis, and control the temperature at 30°C for 5 min.

[0048] (3) The extract was pretreated by 2μm plate and frame filtration. The filtrate was then clarified by a 5nm ceramic membrane under the conditions of 0.1MPa operating pressure, 2m / s membrane flow rate and 10℃ operating temperature. The clarified liquid was dried in an oven at 80℃ to obtain a dry extract, thus obtaining the Senecio scandens extract.

[0049] Example 2: Improved process for Senecio scandens

[0050] (1) Take a certain amount of Senecio scandens, add water at 100℃ and decoct twice, each time for 2 hours, add 10 times the amount of water each time, and add citric acid to adjust the pH to 4. Extract twice, combine the decoctions and set aside.

[0051] (2) Add 1% (v / v) of formaldehyde sulfuric acid to the Senecio scandens extract for catalysis, with the temperature controlled at 60℃ and the time at 15 min.

[0052] (3) The extract was pretreated by 2μm plate and frame filtration. The filtrate was then clarified by a 50nm ceramic membrane under the conditions of operating pressure of 0.5MPa, membrane flow rate of 4m / s and operating temperature of 50℃. The clarified liquid was dried in an oven at 80℃ to obtain a dry extract, thus obtaining the Senecio scandens extract.

[0053] Example 3: Improved process for Senecio scandens

[0054] (1) Take a certain amount of Senecio scandens, add water at 80℃ and decoct twice, each time for 2 hours, add 10 times the amount of water each time, and add citric acid to adjust the pH to 4. Extract twice, combine the decoctions and set aside.

[0055] (2) Add 0.5% (v / v) ammonium molybdate sulfuric acid to the Senecio scandens extract for catalysis, with the temperature controlled at 45℃ and the time at 10 min.

[0056] (3) The extract was pretreated by 2μm plate and frame filtration. The filtrate was then clarified by a 50nm ceramic membrane under the conditions of operating pressure 0.2MPa, membrane flow rate 4m / s and operating temperature 50℃. The clarified liquid was dried in an oven at 80℃ to obtain a dry extract, thus obtaining the Senecio scandens extract.

[0057] Comparative Example 1: Senriko Original Process

[0058] (1) Take a certain amount of Senecio scandens, add water at 100℃ and decoct twice, each time for 2 hours, add 10 times the amount of water each time, extract twice, combine the decoctions and set aside;

[0059] (2) The extract was pretreated by 2μm plate and frame filtration. The filtrate was then clarified by a 50nm ceramic membrane under the conditions of operating pressure 0.2MPa, membrane flow rate 4m / s and operating temperature 50℃. The clarified liquid was dried in an oven at 80℃ to obtain a dry extract, thus obtaining the Senecio scandens extract.

[0060] Example 4: Improved process for Senecio scandens

[0061] (1) Take a certain amount of Senecio scandens, add water at 60℃ and decoct twice, each time for 2 hours, add 10 times the amount of water each time, and add citric acid to adjust the pH to 2. Extract twice, combine the decoctions and set aside.

[0062] (2) Add 1% volume fraction of ammonium molybdate sulfuric acid to the Senecio scandens extract for catalysis, with the temperature controlled at 30℃ and the time at 5min.

[0063] (3) The extract was pretreated by 2μm plate and frame filtration. The filtrate was then clarified by a 35nm ceramic membrane under the conditions of operating pressure 0.1MPa, membrane flow rate 1m / s and operating temperature 10℃. The clarified liquid was dried in an oven at 80℃ to obtain a dry extract, thus obtaining the Senecio scandens extract.

[0064] Example 5: Improved process for Senecio scandens

[0065] (1) Take a certain amount of Senecio scandens, add water at 60℃ and decoct twice, each time for 2 hours, add 10 times the amount of water each time, and add citric acid to adjust the pH to 2. Extract twice, combine the decoctions and set aside.

[0066] (2) Add 0.1% volume fraction of silicotungstic acid to the extract of Senecio scandens for catalysis, and control the temperature at 50℃ for 5 min.

[0067] (3) The extract was pretreated by 2μm plate and frame filtration. The filtrate was then clarified by a 35nm ceramic membrane under the conditions of operating pressure 0.2MPa, membrane flow rate 2m / s and operating temperature 10℃. The clarified liquid was dried in an oven at 80℃ to obtain a dry extract, thus obtaining the Senecio scandens extract.

[0068] Example 6: Improved process for Senecio scandens

[0069] (1) Take a certain amount of Senecio scandens, add water at 60℃ and decoct twice, each time for 2 hours, add 10 times the amount of water each time, and add citric acid to adjust the pH to 2. Extract twice, combine the decoctions and set aside.

[0070] (2) Add 0.1% volume fraction of silicotungstic acid to the extract of Senecio scandens for catalysis, and control the temperature at 30℃ for 15 min.

[0071] (3) The extract was pretreated by 2μm plate and frame filtration. The filtrate was then clarified by a 35nm ceramic membrane under the conditions of operating pressure 0.2MPa, membrane flow rate 2m / s and operating temperature 10℃. The clarified liquid was dried in an oven at 80℃ to obtain a dry extract, thus obtaining the Senecio scandens extract.

[0072] Example 7: Improved process for Senecio scandens

[0073] (1) Take a certain amount of Senecio scandens, add water at 100℃ and decoct twice, each time for 2 hours, add 10 times the amount of water each time, and add citric acid to adjust the pH to 4. Extract twice, combine the decoctions and set aside.

[0074] (2) Add 1% volume fraction of phosphomolybdic acid to the extract of Senecio scandens for catalysis, and control the temperature at 60℃ for 15 min.

[0075] (3) The extract was pretreated by 2μm plate and frame filtration. The filtrate was then clarified by a 35nm ceramic membrane under the conditions of operating pressure 0.3MPa, membrane flow rate 4m / s and operating temperature 50℃. The clarified liquid was dried in an oven at 80℃ to obtain a dry extract, thus obtaining the Senecio scandens extract.

[0076] Example 8

[0077] (1) Take a certain amount of Senecio scandens, add water at 100℃ and decoct twice, one hour each time, add 10 times the amount of water each time, and add citric acid to adjust the pH to 3. Extract twice, combine the decoctions and set aside.

[0078] (2) Add 1% volume fraction of silicotungstic acid to the extract of Senecio scandens for catalysis, and control the temperature at 60℃ for 15 min.

[0079] (3) The extract was pretreated by 10μm plate and frame filtration. The filtrate was then clarified by a 35nm ceramic membrane under the conditions of operating pressure of 0.3MPa, membrane flow rate of 4m / s and operating temperature of 50℃. The clarified liquid was dried in an oven at 80℃ to obtain a dry extract, thus obtaining the Senecio scandens extract.

[0080] The above embodiments and comparative examples were tested, and the HPLC-MS conditions for adoniflunline were as follows: Figure 1 As shown, the determination of different concentrations of adoniflurine (ADNFLJ) standard by the internal standard method using pyruvic alkaloid (YBHJ) was performed by HPLC-MS. Figure 2-5 As shown (comparisons 1-4), the original extract of *Senecio scandens*, the clarified and concentrated solutions after clarification using conventional ceramic membrane methods, and the clarified and concentrated solutions after clarification using inorganic acid-catalyzed ceramic membrane methods are as follows: Figure 6-20 As shown in Table 1, the specific results are as follows.

[0081] Table 1 Concentrations of Adoniflurin, Chlorogenic Acid, and Hyperoside

[0082]

[0083] Note: The original solution is the decoction obtained in step (1) of Comparative Example 1; the conventional process is Comparative Example 1; the improved process is Example 8; the concentrated solution and the clear solution are the ceramic membrane concentrated solution and ceramic membrane clear solution after ceramic membrane treatment, respectively.

[0084] In the improved process, the index components such as chlorogenic acid and hyperoside were not lost, but the adoniflurine content was greatly reduced. Examples 1-7 also showed a significant reduction in adoniflurine.

[0085] In summary, the traditional method for preparing Senecio scandens extract involves pulverizing the herb, extracting with boiling water, filtering with a ceramic membrane, and concentrating with an inorganic membrane. This method cannot effectively retain adorniflunline (PAs), only large-molecule proteins, particles, and impurities. Using the process of this invention, after acid extraction, the adorniflunline (PAs) in the herb can undergo a catalytic reaction with the inorganic acid, forming a complex precipitate. After filtration with a plate and frame filter and a ceramic membrane, adorniflunline is essentially completely removed. Then, an inorganic nanofiltration membrane with a specific pore size is used to allow small-molecule inorganic acids, terpenes, tannins, and other active ingredients to pass through, while retaining potentially incompletely reacted inorganic acids and small-molecule proteins. Finally, the supernatant is concentrated using a reverse osmosis membrane and spray-dried to obtain a Senecio scandens extract that is essentially free of adorniflunline (PAs).

[0086] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are specific and detailed, they should not be construed as limiting the scope of the present invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these modifications and improvements all fall within the scope of protection of the present invention. Therefore, the scope of protection of this patent should be determined by the appended claims.

Claims

1. A method for preparing Senecio scandens extract, characterized in that, Includes the following steps: (1) Extract the first extract of Senecio scandens with water at 60-100℃ under acidic conditions with a pH of 2-5. (2) The first extract obtained was reacted at 20-70℃ under the catalysis of inorganic acid to obtain the second extract; (3) The second extract obtained is filtered through a plate and frame filter with a pore size of 0.5-15μm and a ceramic membrane filter. The filtrate is dried to obtain a Senecio scandens extract with an adoniflunine content of less than 0.001%. In step (2), the inorganic acid is any one or a combination of formaldehyde sulfuric acid, ammonium molybdate sulfuric acid, silicotungstic acid and phosphomolybdic acid; the amount of the inorganic acid used is 0.1%-1% of the volume or mass of the first extract. In step (3), the pore size of the ceramic membrane is 5-50 nm; the pressure of the ceramic membrane filtration is 0.1-0.5 MPa; and the flow rate of the ceramic membrane filtration surface is 1-4 m / s.

2. The method according to claim 1, characterized in that, In step (1), the acidic conditions are any one or more combinations of hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid and citric acid.

3. The method according to claim 1, characterized in that, In step (2), the formaldehyde sulfuric acid is a combination of formaldehyde and sulfuric acid, with a volume ratio of formaldehyde to sulfuric acid of 1:5-7; the ammonium molybdate sulfuric acid is a combination of ammonium molybdate and sulfuric acid, with a volume ratio of ammonium molybdate to sulfuric acid of 1:95-103.

4. The method according to claim 1, characterized in that, In step (2), the formaldehyde sulfuric acid is a combination of formaldehyde and sulfuric acid, with a volume ratio of 1:6; the ammonium molybdate sulfuric acid is a combination of ammonium molybdate and sulfuric acid, with a volume ratio of 1:

99.

5. The method according to claim 1, characterized in that, In step (2), the reaction temperature is 30-60℃.

6. The method according to claim 1, characterized in that, In step (2), the reaction time is 3-20 min.

7. The method according to claim 1, characterized in that, In step (2), the reaction time is preferably 5-15 min.

8. The method according to claim 1, characterized in that, In step (3), the aperture of the plate frame is 1-10 μm.

9. The method according to claim 1, characterized in that, In step (3), the aperture of the plate frame is 2-10 μm.

10. The method according to claim 1, characterized in that, In step (3), the temperature of the ceramic membrane filtration is 20-50℃.

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