Processing technology for weakening toxicity of polygonum multiflorum
Through soaking rice water and steaming black bean juice to combine enzyme solution, the hepatic toxicity problem caused by styrene glycoside and binding anthraquinone in the preparation of Polygonum multiflorum was solved, the process was simplified, the toxicity was reduced, the efficacy was improved, and the pharmacopoeia standards were met.
Patent Information
- Application Number
- CN202411290212.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-09-14
- Publication Date
- 2024-12-13
AI Technical Summary
In the existing process of polypsi, the hepatotoxicity problems caused by styrene glycoside and anthraquinone are caused by long-term use, which may cause liver damage. The traditional process is cumbersome and time-consuming, making it difficult to effectively control toxicity and improve the efficacy.
Soak the Polygonum multiflorum tablets in rice water and mix well with black bean juice to steam it. The enzyme solution combining cellulase and β-glucosidase is soaked to promote the reduction of styrene glycoside and the conversion of anthraquinone, and reduce hepatotoxicity.
The preparation process is simplified, the time is shortened, the styrene glycoside content is reduced, the free anthraquinone content is increased, the hepatotoxic reaction is avoided, the Chinese pharmacopoeia standards is met, and the safety and efficacy of Polygonum multiflorum are improved.
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Abstract
Description
Technical Field
[0001] The invention relates to the field of traditional Chinese medicine processing, and in particular to a processing technology for reducing the toxicity of Polygonum multiflorum. Background Art
[0002] Polygonum multiflorum is the dried root of Polygonum multiflorum Thunb. of the Polygonaceae family. It tastes bitter, sweet, astringent, is slightly warm in nature, and enters the liver, heart, and kidney meridians. When used raw, Polygonum multiflorum has the effects of detoxification, eliminating carbuncle, stopping malaria, moistening the intestines and relieving constipation. After processing, it has the effects of nourishing the liver and kidneys, replenishing the essence and blood, strengthening the tendons and bones, blackening the hair, clearing turbidity and reducing lipids. Modern pharmacological studies have shown that Polygonum multiflorum has pharmacological activities such as antioxidant, anti-tumor, anti-atherosclerosis, anti-aging, immunity enhancement, blood lipid lowering and neuroprotection, and has the potential to treat neurodegenerative diseases and lower blood sugar. The effects of Polygonum multiflorum can be summarized as "raw purgation and cooked tonification". It can be seen that the processing technology has a great influence on the chemical composition and pharmacological activity of Polygonum multiflorum.
[0003] Raw Polygonum multiflorum is toxic. It is used clinically after processing. After processing and compatibility, the toxicity of processed Polygonum multiflorum is greatly reduced and the efficacy is increased. There are many processing methods for Polygonum multiflorum, and more than a dozen auxiliary materials are used in the processing, including black beans, rice wine, black sesame, vinegar, rice water, etc., among which black beans are the most processed; the processing technology is mainly "steaming", among which "nine steaming and nine drying" is the mainstream processing method of Polygonum multiflorum, which has been used since the Song Dynasty. It is also the most complicated and extremely time-consuming processing method. However, a large number of clinical reports in recent years have shown that long-term use of processed Polygonum multiflorum and its preparations can cause certain liver damage, mainly idiosyncratic hepatotoxicity. At present, some studies have found that diphenylethylene glycosides are the main chemical components of the idiosyncratic hepatotoxicity of Polygonum multiflorum, and the inflammatory response caused by trans-diphenylethylene glycosides can further induce the idiosyncratic hepatotoxicity of cis-diphenylethylene glycosides; some studies have also found that the combined anthraquinone components have a strong competitive inhibitory effect on the UGT1A1-mediated bilirubin metabolites, resulting in bilirubin unable to be converted into water-soluble bilirubin gluconate conjugates and excreted from the body, resulting in metabolic disorders, which accumulate in the liver blood, and severe cases have hepatotoxic reactions. Therefore, it is urgent to develop a processing technology that can not only make the effective content of processed Polygonum multiflorum reach the Chinese Pharmacopoeia standard, but also effectively control the diphenylethylene glycoside content, promote the conversion of combined anthraquinones to free anthraquinones, and effectively reduce the hepatotoxicity of processed Polygonum multiflorum. Summary of the invention
[0004] 1. Technical issues to be resolved
[0005] In view of the deficiencies in the prior art, the present invention provides a processing technology for reducing the toxicity of Polygonum multiflorum.
[0006] (II) Technical solution
[0007] To achieve the above objectives, the present invention is implemented through the following technical solutions:
[0008] The present invention provides a processing process for reducing the toxicity of Polygonum multiflorum, comprising the following steps:
[0009] (1) Clean and slice Polygonum multiflorum, soak it in rice water overnight, and then dry it in an oven;
[0010] (2) Weigh black beans, add appropriate amount of water, boil for 4 hours, and filter to obtain black bean juice and bean dregs; add appropriate amount of water to the bean dregs and boil again for 3 hours, filter, and combine the filtrate to obtain black bean juice;
[0011] (3) Mix the Polygonum multiflorum slices and black bean juice, steam them in a pressure cooker, and then dry them in an oven;
[0012] (4) Soak the Polygonum multiflorum slices in the enzyme solution, take them out, and bake them in an oven for 8-10 hours to obtain the processed Polygonum multiflorum.
[0013] Specifically, in the processing process, 10 kg of black beans are needed for every 100 kg of Polygonum multiflorum slices, and 25 kg of black bean juice can be boiled for every 10 kg of black beans.
[0014] Specifically, in the processing step (3), the steaming time of the Polygonum multiflorum slices in the pressure cooker is 5-8 hours.
[0015] Specifically, the enzyme solution in the processing step (4) is an aqueous solution containing at least two enzymes of cellulase and β-glucosidase.
[0016] Specifically, in the preparation process step (4), the ratio of cellulase to β-glucosidase in the enzyme solution is 1:1-4.
[0017] Specifically, the ratio of cellulase to β-glucosidase in the enzyme solution of the preparation process step (4) is 1:4.
[0018] Specifically, the total amount of cellulase and β-glucosidase used in the processing step (4) is 0.1-1% of the amount of Polygonum multiflorum used.
[0019] Specifically, the total amount of cellulase and β-glucosidase used in the processing step (4) is 0.1% of the amount of Polygonum multiflorum used.
[0020] Specifically, in the processing step (4), the soaking time of the Polygonum multiflorum slices in the enzyme solution is 8-12 hours.
[0021] Specifically, in the preparation process, the oven temperature in step (1) and step (3) is 75-80°C, and the oven temperature in step (4) is 40-45°C.
[0022] (III) Beneficial effects
[0023] The invention provides a processing process for reducing the toxicity of Polygonum multiflorum. The process comprises the following steps: firstly soaking Polygonum multiflorum in rice water to reduce its pungent and dry smell and enhance the effect of invigorating the spleen and harmonizing the middle part; then mixing with black bean juice and steaming to enhance the effect of nourishing the liver and kidney and blackening the hair; enzyme is an important biological catalyst with high specificity to substrates; cellulase is a composite enzyme composed of exo-β-glucanase, endo-β-glucanase and β-glucosidase; β-glucosidase is also called β-D-glucosidase hydrolase and can hydrolyze β-D-glucose bond bound to terminal non-reducing part and release β-glucose and corresponding ligand at the same time; the two enzymes act synergistically to control the content of diphenylethylene glycoside in Polygonum multiflorum and promote the conversion of bound anthraquinone to free anthraquinone. Compared with the prior art, the processing steps provided by the present invention are simple, greatly shortening the processing time of traditional processed Polygonum multiflorum, and the addition of cellulase and beta-glucosidase can reduce the loss of effective ingredients, while increasing the dissolution and conversion of toxic ingredients, thereby avoiding idiosyncratic hepatotoxicity caused by long-term use of processed Polygonum multiflorum. The processed Polygonum multiflorum obtained by the processing process provided by the present invention meets the standards of the Chinese Pharmacopoeia. DETAILED DESCRIPTION
[0024] In order to make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution in the embodiments of the present invention will be clearly and completely described below in combination with the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.
[0025] Example 1
[0026] A processing process for reducing the toxicity of Polygonum multiflorum comprises the following steps:
[0027] (1) Weigh 10 kg of Polygonum multiflorum, wash it, slice it, soak it in rice water overnight, and dry it in an oven at 75°C;
[0028] (2) Weigh 1 kg of black beans, add 8 L of water, boil for 4 h, and filter to obtain black bean juice and dregs; add 6 L of water to the dregs and boil again for 3 h, filter, and combine the filtrate to obtain black bean juice;
[0029] (3) Mix the Polygonum multiflorum slices and black bean juice, steam them in a pressure cooker for 5 h, and dry them in an oven at 75°C;
[0030] (4) Weigh 2 g of cellulase and 8 g of β-glucosidase and dissolve them in distilled water to obtain a mixed enzyme solution of cellulase and β-glucosidase. Soak the Polygonum multiflorum slices in the mixed enzyme solution for 10 h and bake them in an oven at 40° C. for 10 h to obtain the processed Polygonum multiflorum.
[0031] Example 2
[0032] A processing process for reducing the toxicity of Polygonum multiflorum comprises the following steps:
[0033] (1) Weigh 10 kg of Polygonum multiflorum, wash it, slice it, soak it in rice water overnight, and dry it in an oven at 75°C;
[0034] (2) Weigh 1 kg of black beans, add 8 L of water, boil for 4 h, and filter to obtain black bean juice and dregs; add 6 L of water to the dregs and boil again for 3 h, filter, and combine the filtrate to obtain black bean juice;
[0035] (3) Mix the Polygonum multiflorum slices and black bean juice, steam them in a pressure cooker for 8 h, and dry them in an oven at 75°C;
[0036] (4) Weigh 2 g of cellulase and 8 g of β-glucosidase and dissolve them in distilled water to obtain a mixed enzyme solution of cellulase and β-glucosidase. Soak the Polygonum multiflorum slices in the mixed enzyme solution for 8 h and bake them in an oven at 40° C. for 10 h to obtain the processed Polygonum multiflorum.
[0037] Example 3
[0038] A processing process for reducing the toxicity of Polygonum multiflorum comprises the following steps:
[0039] (1) Weigh 10 kg of Polygonum multiflorum, wash it, slice it, soak it in rice water overnight, and dry it in an oven at 75°C;
[0040] (2) Weigh 1 kg of black beans, add 8 L of water, boil for 4 h, and filter to obtain black bean juice and dregs; add 6 L of water to the dregs and boil again for 3 h, filter, and combine the filtrate to obtain black bean juice;
[0041] (3) Mix the Polygonum multiflorum slices and black bean juice, steam them in a pressure cooker for 6 h, and dry them in an oven at 75°C;
[0042] (4) Weigh 2 g of cellulase and 8 g of β-glucosidase and dissolve them in distilled water to obtain a mixed enzyme solution of cellulase and β-glucosidase. Soak the Polygonum multiflorum slices in the mixed enzyme solution for 9 h and bake them in an oven at 40° C. for 10 h to obtain the processed Polygonum multiflorum.
[0043] Example 4
[0044] A processing process for reducing the toxicity of Polygonum multiflorum comprises the following steps:
[0045] (1) Weigh 10 kg of Polygonum multiflorum, wash it, slice it, soak it in rice water overnight, and dry it in an oven at 75°C;
[0046] (2) Weigh 1 kg of black beans, add 8 L of water, boil for 4 h, and filter to obtain black bean juice and dregs; add 6 L of water to the dregs and boil again for 3 h, filter, and combine the filtrate to obtain black bean juice;
[0047] (3) Mix the Polygonum multiflorum slices and black bean juice, steam them in a pressure cooker for 5 h, and dry them in an oven at 75°C;
[0048] (4) Weigh 4 g of cellulase and 6 g of β-glucosidase and dissolve them in distilled water to obtain a mixed enzyme solution of cellulase and β-glucosidase. Soak the Polygonum multiflorum slices in the mixed enzyme solution for 10 h and bake them in an oven at 40° C. for 10 h to obtain the processed Polygonum multiflorum.
[0049] Example 5
[0050] A processing process for reducing the toxicity of Polygonum multiflorum comprises the following steps:
[0051] (1) Weigh 10 kg of Polygonum multiflorum, wash it, slice it, soak it in rice water overnight, and dry it in an oven at 75°C;
[0052] (2) Weigh 1 kg of black beans, add 8 L of water, boil for 4 h, and filter to obtain black bean juice and dregs; add 6 L of water to the dregs and boil again for 3 h, filter, and combine the filtrate to obtain black bean juice;
[0053] (3) Mix the Polygonum multiflorum slices and black bean juice, steam them in a pressure cooker for 5 h, and dry them in an oven at 75°C;
[0054] (4) Weigh 3 g of cellulase and 7 g of β-glucosidase and dissolve them in distilled water to obtain a mixed enzyme solution of cellulase and β-glucosidase. Soak the Polygonum multiflorum slices in the mixed enzyme solution for 10 h and bake them in an oven at 40° C. for 10 h to obtain the processed Polygonum multiflorum.
[0055] Test example
[0056] 1. Materials
[0057] The processed Polygonum multiflorum used in the control group was purchased from Zhang Zhongjing Pharmacy.
[0058] 2,3,5,4′-Tetrahydroxystilbene-2-O-β-D-glucoside reference substance was purchased from Shanghai Yuanye Biotechnology Co., Ltd.;
[0059] Rhamnosin reference substance and Rhamnosin methyl ether reference substance were purchased from Chengdu Pusi Biotechnology Co., Ltd.
[0060] 2. Methods
[0061] Chromatographic conditions
[0062] Chromatographic column: Agilent Zorbox SB-C18 (250×4.6mm, 5μm); mobile phase: water (A)-acetonitrile (B); gradient elution: 0-15min: 10%-30% B; 15-30min: 30%-45% B; 30-35min: 45%-80% B; 35-40min: 80%-90% B; 40-50min: 90%-10% B; 50-55min: 10% B; column temperature: 30℃; flow rate: 1mL / min; detection wavelength: 254nm; injection volume: 10μL.
[0063] Solution preparation
[0064] Reference solution: Take an appropriate amount of 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside reference substance, accurately weigh it, and add methanol to make a 0.2 mg / mL solution to obtain 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside reference substance solution;
[0065] Take an appropriate amount of rheum officinale reference substance, weigh it accurately, and add methanol to make a solution of 80 μg / mL to obtain rheum officinale reference substance solution;
[0066] Take an appropriate amount of physaccharin methyl ether reference substance, weigh it accurately, and add methanol to make a 40 μg / mL solution to obtain the physaccharin methyl ether reference substance solution;
[0067] Test solution: Take 0.2 g of prepared Polygonum multiflorum powder (passed through No. 4 sieve), weigh accurately, place in a stoppered conical flask, add 25 mL of methanol accurately, weigh the weight, heat and reflux for 30 minutes, take out and cool, weigh again, make up the lost weight with methanol, shake well, let stand, filter the supernatant, and take the filtrate to obtain.
[0068] 2.3. Content determination
[0069] Prepare test solutions of prepared Polygonum multiflorum obtained according to different processing techniques according to the method described in 2.2 above, and determine the content under the chromatographic conditions described in 2.1 above, and calculate the contents of stilbene glycosides (calculated as 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside) and free anthraquinone (calculated as the total amount of rhamnolide and rhamnolide methyl ether).
[0070] Statistical methods
[0071] The data were processed using GraphPad Prism 8 software and the results are presented as mean The data are expressed as ± standard deviation (s), and one-way analysis of variance was used for comparison among groups.
[0072] Process optimization
[0073] 2.5.1. Effect of enzyme types on the content of active ingredients in processed Polygonum multiflorum
[0074] We first studied the composition of the enzyme solution in the process of the present invention. The compositions of different enzyme solutions are shown in Table 1. The total amount of enzyme used is 10 g, the ratio is 1:1:1, and the other preparation conditions are the same as in Example 1.
[0075] Table 1 Enzyme addition
[0076] Grouping Cellulase β-Glucosidase Amylase Test Group 1 + - - Test Group 2 - + - Experiment 3 groups - - + Test 4 groups + + - Test 5 groups + - + Experiment 6 groups - + + Experiment 7 groups + + +
[0077] Note: + indicates that the enzyme was added, - indicates that the enzyme was not added.
[0078] The effects of enzyme solutions with different components on the contents of stilbene glycosides and free anthraquinones in processed Polygonum multiflorum are shown in Table 2. Compared with the control group, the stilbene content in processed Polygonum multiflorum in each group was significantly reduced, and the free anthraquinone content was significantly increased, with significant differences (P<0.05), and all met the standards of the Chinese Pharmacopoeia. Among them, the effect of the mixed use of cellulase and β-glucosidase in test group 4 was significantly better than that of test groups 1-3 and test groups 5-7, with significant differences (P<0.05), indicating that the synergistic effect of cellulase and β-glucosidase is better than the use of a single enzyme or a mixture with amylase.
[0079] Table 2 The content of effective ingredients in each experimental group of Polygonum multiflorum
[0080]
[0081] Data in the same column with different letters indicate significant differences (P<0.05), and data with the same letters indicate no significant differences (P>0.05).
[0082] 2.5.2. Effect of enzyme ratio on the content of active ingredients in processed Polygonum multiflorum
[0083] On the basis of determining that the enzyme solution components are cellulase and β-glucosidase, we studied the ratio relationship between the two enzymes. Different ratios of cellulase and β-glucosidase are shown in Table 3. The other preparation conditions are the same as in Example 1.
[0084] Table 3 Proportions of enzyme solution
[0085]
[0086] The results are shown in Table 4. Compared with the control group, the content of diphenylethylene in the processed Polygonum multiflorum in each group was significantly reduced (P<0.05), and the content of free anthraquinone was significantly increased, and all met the standards of the Chinese Pharmacopoeia. Among them, the effect of Experiment 2 was significantly better than that of Experiment 1 and Experiments 3-5, and the difference was significant (P<0.05). Therefore, the optimal ratio of cellulase to β-glucosidase in the processing process provided by the present invention is 1:4.
[0087] Table 4 The content of effective ingredients in each experimental group of Polygonum multiflorum
[0088]
[0089] Data in the same column with different letters indicate significant differences (P<0.05), and data with the same letters indicate no significant differences (P>0.05).
[0090] 2.5.3. Effect of processing time on the content of active ingredients in processed Polygonum multiflorum
[0091] We further studied the effects of pressure cooker steaming time and enzyme solution soaking time on the content of effective ingredients in processed Polygonum multiflorum. Different pressure cooker steaming times and enzyme solution soaking times are shown in Table 5. The other processing conditions are the same as in Example 1.
[0092] Table 5 Pressure cooker steaming time and enzyme solution soaking time
[0093]
[0094] The results are shown in Table 6. Compared with the control group, the content of stilbene in each group of processed Polygonum multiflorum was significantly reduced, and the content of free anthraquinone was significantly increased, with significant differences (P<0.05), and all met the standards of the Chinese Pharmacopoeia. Among them, the content of stilbene glycosides in the test group 2 was the lowest, and the content of free anthraquinone was the highest, which was significantly different from the other test groups (P<0.05). Therefore, the optimal processing time of Polygonum multiflorum in the processing process provided by the present invention is to steam it in a pressure cooker for 5 hours first, and then soak it in the enzyme solution for 10 hours.
[0095] Table 6 The content of effective ingredients in each experimental group of Polygonum multiflorum
[0096]
[0097]
[0098] Data in the same column with different letters indicate significant differences (P<0.05), and data with the same letters indicate no significant differences (P>0.05).
[0099] Process validation
[0100] Take an appropriate amount of Polygonum multiflorum, wash it, slice it, divide it into 3 parts, and process it according to the above process optimization results to determine the contents of diphenylethylene glycoside and anthraquinone in the processed Polygonum multiflorum.
[0101] The results are shown in Table 7, indicating that the preparation process is stable and feasible with good repeatability.
[0102] Table 7 Verification test results (n=3)
[0103]
[0104] 3. Discussion
[0105] The present invention optimizes the processing technology of Polygonum multiflorum by single factor test with stilbene glycoside and free anthraquinone content as evaluation criteria, and determines that the best processing technology of Polygonum multiflorum is to soak Polygonum multiflorum in rice water overnight, mix it with black bean juice, put it in a pressure cooker for steaming for 5 hours and then dry it, then put it in a mixed enzyme solution of cellulase and beta-glucosidase for soaking for 10 hours and then bake it for 8-10 hours to obtain the processed Polygonum multiflorum, wherein the ratio of cellulase to beta-glucosidase is 1:4, and the total amount is 0.1-1% of the amount of Polygonum multiflorum. The stilbene content in the processed Polygonum multiflorum obtained by the processing technology of the present invention is significantly reduced within the effective range, and the free anthraquinone content is significantly increased, which can avoid the idiosyncratic hepatotoxicity caused by long-term use of Polygonum multiflorum. Compared with the traditional preparation technology of processed Polygonum multiflorum, the processing technology of the present invention is simplified, the processing time is greatly shortened, the efficiency is high, the obtained processed Polygonum multiflorum is of good quality, and has good economic benefits.
[0106] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit the same. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that the technical solutions described in the aforementioned embodiments may still be modified, or some of the technical features may be replaced by equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.
Claims
1. A processing technology for reducing the toxicity of Polygonum multiflorum, It is characterized in that The following steps are involved: (1) Clean and slice Polygonum multiflorum, soak it in rice water overnight, and then dry it in an oven; (2) Weigh black beans, add appropriate amount of water, boil for 4 hours, and filter to obtain black bean juice and bean dregs; add appropriate amount of water to the bean dregs and boil again for 3 hours, filter, and combine the filtrate to obtain black bean juice; (3) Mix the Polygonum multiflorum slices and black bean juice, steam them in a pressure cooker, and dry them in an oven; (4) Soak the Polygonum multiflorum slices in the enzyme solution, take them out, and bake them in an oven for 8-10 hours to obtain the processed Polygonum multiflorum.
2. A processing process for reducing the toxicity of Polygonum multiflorum according to claim 1, It is characterized in that In the step (3), the steaming time of the Polygonum multiflorum slices in the pressure cooker is 5-8 hours.
3. A processing process for reducing the toxicity of Polygonum multiflorum according to claim 1, It is characterized in that The enzyme solution in step (4) is an aqueous solution containing at least two enzymes of cellulase and β-glucosidase.
4. The enzyme solution as claimed in claim 3, wherein the ratio of cellulase to β-glucosidase is 1:1-4, and the total dosage is 0.1-1% of the dosage of Polygonum multiflorum.
5. The processing process for reducing the toxicity of Polygonum multiflorum according to claim 1, It is characterized in that In the step (4), the soaking time of the Polygonum multiflorum slices in the enzyme solution is 8-12 hours.
6. The processing process for reducing the toxicity of Polygonum multiflorum according to claim 1, It is characterized in that The temperature of the oven in step (1) and step (3) is 75-80°C, and the temperature of the oven in step (4) is 40-45°C.