Angelica sinensis, paeonia lactiflora and rehmannia pill and preparation method thereof

Abstract

The invention relates to the technical field of preparation of angelica sinensis, paeonia lactiflora and rehmannia pills, and discloses angelica sinensis, paeonia lactiflora and rehmannia pills and a preparation method thereof. 35-45 parts of wine-processed radix paeoniae alba; 150 to 170 parts of prepared rehmannia root; 75 to 85 parts of wine-processed fructus corni; 55 to 65 parts of cortex moutan; 75 to 85 parts of Chinese yam; 55 to 65 parts of poria cocos; 55 to 65 parts of rhizoma alismatis; each part of the pill contains 5-20mg of vitamin C and 2-10mg of vitamin E, and the addition ratio of the vitamin C to the vitamin E is 3: 1. Vitamin C and vitamin E are added into the angelica sinensis, paeonia lactiflora and rehmannia pill to form an innovative formula which takes traditional Chinese medicines as a main body and combines with novel auxiliary materials, and the synergistic anti-oxidation effect of vitamin C and vitamin E is utilized, so that active ingredients in the pill are effectively protected from being oxidized or degraded, and the immune regulation function is enhanced.

Description

Technical Field

[0001] The invention relates to the technical field of Guishao Dihuang Pill preparation, in particular to Guishao Dihuang Pill and a preparation method thereof. Background Art

[0002] Guishao Dihuang Wan is one of the classic prescriptions of traditional Chinese medicine. It has the effects of nourishing yin and kidney, nourishing blood and regulating menstruation. It is widely used in the conditioning of low immune function and the improvement of sub-health status. However, the existing preparation method mainly relies on the traditional water extraction and alcohol precipitation process, and the extraction efficiency of active ingredients in medicinal materials (such as catalpol, paeoniflorin, polysaccharides, etc.) is low. In particular, some heat-sensitive ingredients are easily decomposed or degraded under high temperature conditions, resulting in the loss of effective ingredients of the drug. In addition, due to the large difference in polarity of active ingredients, it is difficult to achieve targeted extraction of multiple ingredients with traditional processes, resulting in significant deficiencies in product efficacy. How to improve the extraction rate and retention rate of active ingredients by optimizing extraction technology has become an urgent problem to be solved.

[0003] In terms of formulation stability, the existing process of Guishao Dihuang Pills is relatively extensive, and the active ingredients in the product are easily oxidized or degraded during storage, resulting in attenuation of efficacy. Although traditional excipients such as refined honey can delay the degradation of ingredients to a certain extent, they have limited effects in protecting easily oxidized ingredients and improving the stability of pills. At the same time, the lack of precise parameter control of the extraction, concentration and molding links during the preparation process can easily lead to inconsistent quality between batches. Especially in the purification of the extract, the conventional concentration and coarse filtration methods in the prior art are difficult to effectively remove impurities and enrich key ingredients, further limiting the efficacy and market competitiveness of the preparation.

[0004] In addition, although the traditional formula of Guishao Dihuang Pills can play a certain role in immunomodulation, its antioxidant capacity and potential for improving efficacy are insufficient. Studies have shown that antioxidant capacity is of great significance for enhancing immune function, while the antioxidant capacity of traditional Chinese medicine itself is relatively limited. In the modern Chinese medicine preparation process, the introduction of functional excipients is an effective way to enhance efficacy and product competitiveness, but Guishao Dihuang Pills have not yet fully combined the synergistic advantages of functional ingredients such as vitamins. Summary of the invention

[0005] In view of the deficiencies in the prior art, the present invention provides a Guishao Dihuang Pill and a preparation method thereof, which solves the problem that the existing Guishao Dihuang Pill has a relatively extensive process and the active ingredients in the product are easily oxidized or degraded during storage, resulting in a decrease in efficacy.

[0006] To achieve the above objectives, the present invention is implemented through the following technical scheme: A Guishao Dihuang Pill, comprising the following components in parts by weight: Angelica: 35-45 parts; White peony root with wine: 35-45 parts; Rehmannia root: 150-170 parts; Wine-boiled cornus: 75-85 portions; Peony bark: 55-65 parts; Chinese yam: 75-85 servings; Poria: 55-65 parts; Alisma orientalis: 55-65 parts; Each pill contains 5-20 mg of vitamin C and 2-10 mg of vitamin E, and the pills are prepared by the following process: The process includes the following steps: pretreatment of medicinal materials: cleaning, slicing and drying of medicinal materials; multi-stage temperature-controlled extraction of active ingredients; concentration and purification of extracts; addition and mixing of auxiliary materials; and preparation molding.

[0007] Furthermore, the synergistic effects of Chinese herbal medicine components: The Rehmannia root, white peony root and angelica root in the formula are all classic medicinal materials for nourishing yin and blood, and tonifying the kidney. They have the functions of enhancing immunity and regulating endocrine; Paeonia suffruticosa and Cornus officinalis further balance the overall effect by clearing away heat, nourishing yin, and consolidating the body; Chinese yam, Poria cocos and Alisma orientalis play the role of strengthening the spleen and replenishing qi, promoting absorption, and improving the overall utilization rate of the drug.

[0008] Each component achieves the comprehensive effect of enhancing immune function through multi-target and multi-pathway pharmacological actions.

[0009] Functions of new materials: Vitamin C: It has significant antioxidant function, can remove free radicals in the body, protect the stability of active ingredients of traditional Chinese medicine in the body, promote the synthesis of collagen, and enhance the body's immune response.

[0010] Vitamin E: It is a fat-soluble antioxidant that can protect the cell membrane structure, enhance the body's antioxidant capacity, regulate cellular immune function, and work synergistically with vitamin C to enhance the overall efficacy.

[0011] The addition of vitamins C and E can not only enhance the immunomodulatory effect of the preparation, but also protect the stability of the active ingredients of traditional Chinese medicine during the preparation process and extend the shelf life of the product.

[0012] Preferably, the added ratio of vitamin C and vitamin E is 3:1.

[0013] Furthermore, experiments have shown that when the ratio of vitamin C to vitamin E is 3:1 (such as adding 15 mg of vitamin C and 5 mg of vitamin E to each pill), vitamin C and E form the best synergistic effect in terms of antioxidant and immune regulation. Vitamin C plays a role in rapidly scavenging free radicals in body fluids, while vitamin E protects cell membranes in a fat-soluble environment. Under this ratio, not only can the antioxidant effects of both be maximized, but also the gastrointestinal discomfort that may be caused by excessive vitamin C can be reduced, and the dosage safety can be optimized.

[0014] Preferably, the pills are water-honeyed pills, which are prepared by adding 35-50 g of refined honey to every 100 g of medicinal powder.

[0015] Preferably, the pills are small honey pills, which are prepared by adding 80-110 g refined honey to every 100 g medicinal powder.

[0016] Furthermore, the choice of refined honey: refined honey is a traditional Chinese medicine pill-making auxiliary material, which plays the role of bonding and delaying drug release. It also has certain nourishing effects and can complement the main medicinal effects of Guishao Dihuang Pills.

[0017] Water-honey pills are suitable for patients who need quick effects. Honey pills have a better taste and are easy to take. Small honey pills are more suitable for long-term use due to their higher honey content and are conducive to the preservation of the drug.

[0018] The following steps are involved: Wash the herbs and slice them into slices, the thickness of slices of Radix Rehmanniae glutinosae is 0.5-2.0 mm, and the thickness of slices of other herbs is 1.0-2.0 mm; The multi-stage temperature-controlled extraction was carried out under the following conditions: the temperature of the first stage was 50-55°C, the extraction time was 1.5-2 hours, and the pH was adjusted to 4.0-4.5; the temperature of the second stage was 65-75°C, the extraction time was 2-3 hours, and the pH was adjusted to 5.5-6.5; the temperature of the third stage was 85-95°C, and the extraction time was 1-1.5 hours; The extracts are combined and concentrated to a solid content of 60 to 80 mg / mL, and purified by ultrafiltration and nanofiltration membranes; Add vitamin C and vitamin E, spray dry, prepare powder, and mix with refined honey to make pills; Preparation molding: The mixed liquid is spray-dried to form powder, mixed with refined honey to form pills, and dried to obtain the finished product.

[0019] Furthermore, low-temperature drying can avoid the loss of heat-sensitive components, such as catalpol and paeoniflorin in Rehmannia glutinosa. Slicing can increase the surface area of ​​the medicinal material and improve the extraction efficiency. Multi-stage temperature-controlled extraction can maximize the retention of multiple components by separating the release of active ingredients at different temperatures: In the first stage, heat-sensitive components (such as paeoniflorin and ferulic acid) are extracted, and the pH is adjusted to acidic to reduce component degradation; The second stage releases the polar components (e.g., paeonol and loganin); In the third stage, high temperature is used to extract heat-resistant macromolecules (such as Rehmannia glutinosa polysaccharides), and high temperature is used to promote their dissolution. Vacuum decompression and concentration can reduce the damage of high temperature to the ingredients. Ultrafiltration and nanofiltration can efficiently remove impurities while retaining the effective ingredients to ensure product purity and efficacy. Vitamin C and E are evenly dispersed in a solution state to avoid fluctuations in efficacy caused by uneven addition. Spray drying can quickly convert the concentrate into a uniform powder to avoid degradation of active ingredients caused by long-term high temperature treatment.

[0020] Preferably, the thickness of the medicinal material slices is 0.5 to 1.0 mm, the drying temperature is 40 to 50° C., and the drying time is 8 to 12 hours.

[0021] Preferably, during the first stage of extraction, the weight-to-volume ratio of the medicinal material to water is 1:10 to 1:12.

[0022] Preferably, the concentration step is carried out at a reduced pressure of 0.08 to 0.1 MPa and a concentration temperature of 45 to 55°C.

[0023] Preferably, the spray drying temperature ranges from 70 to 90° C., and the spray pressure ranges from 0.2 to 0.5 MPa.

[0024] Preferably, the refined honey is added in a ratio of 35 to 110 g per 100 g of medicinal material powder during the pill making process.

[0025] Furthermore, the thickness of the medicinal material and the drying conditions directly affect the extraction efficiency. Slicing increases the surface area, while low-temperature drying avoids the loss of heat-sensitive components; Controlling the ratio of medicinal materials to water can improve the solvent's extraction capacity and ensure uniform concentration of the extract; The reduced pressure concentration conditions effectively reduce the risk of thermal degradation of ingredients while concentrating the active ingredients; Optimization of drying parameters ensures uniform powder particles and avoids high temperature damage; The proportion of refined honey affects the uniformity and formability of pill making, providing a guarantee for the stability of the preparation.

[0026] The present invention provides a Guishao Dihuang Pill and a preparation method thereof, which has the following beneficial effects: 1. The present invention adds vitamin C and vitamin E to Guishao Dihuang Pills to form an innovative formula with traditional Chinese medicine as the main body and new excipients, and utilizes the synergistic antioxidant effect of vitamins C and E to effectively protect the active ingredients in the pills from oxidation or degradation, thereby enhancing the immune regulation function. Compared with the existing traditional scheme that only relies on the compatibility of Chinese medicine itself, the present invention breaks through the bottleneck of insufficient antioxidant capacity of Chinese medicine ingredients and significantly improves the overall efficacy of the drug.

[0027] 2. The present invention adopts a multi-stage temperature-controlled extraction process, optimizes the extraction efficiency of active ingredients according to different temperature conditions, and extracts in low, medium and high temperature stages, accurately retaining the key components of different medicinal materials such as catalpol, paeoniflorin and polysaccharides. In traditional preparation methods, the problem of loss of heat-sensitive components due to single high-temperature extraction is relatively common. The improvement of the present invention avoids these problems, significantly improves the extraction rate of effective ingredients, and the technical solution has higher controllability.

[0028] 3. The present invention uses a combination of ultrafiltration membrane and nanofiltration membrane technology to achieve effective removal of impurities and large molecular weight ineffective components, while enriching active ingredients with moderate molecular weight. The problems of low purity of conventional extracts after concentration and poor drug stability in the prior art are solved. The purification technology scheme of the present invention not only improves the purity of the preparation, but also extends the shelf life of the product, providing a feasible path for large-scale production.

[0029] 4. The present invention uses a combination of ultrafiltration membrane and nanofiltration membrane technology to achieve effective removal of impurities and large molecular ineffective components, while enriching active ingredients with moderate molecular weight. The problems of low purity of conventional extracts after concentration and poor drug stability in the prior art are solved. The purification technology solution of the present invention not only improves the purity of the preparation, but also extends the shelf life of the product, providing a feasible path for large-scale production. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 The present invention is a flow chart of the method. DETAILED DESCRIPTION

[0031] The following will be combined with the drawings in the specification of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only 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.

[0032] Please see attached Figure 1 : Example

[0033] Preparation of Guishao Dihuang Pills and its basic process parameters Pretreatment of medicinal materials Weigh 40g of Angelica sinensis, 40g of white peony root with wine, 160g of Rehmannia glutinosa, 80g of Cornus officinalis with wine, 60g of Paeonia suffruticosa, 80g of Dioscorea opposita, 60g of Poria cocos, and 60g of Alisma orientalis.

[0034] The medicinal materials were rinsed with deionized water (temperature 40°C) to remove surface impurities.

[0035] The slices of Radix Rehmanniae were cut to a thickness of 0.5 mm, and the slices of other herbs were cut to a thickness of 1 mm. After slicing, they were placed in a drying oven at 45°C for 10 hours to ensure that the moisture content of the herbs was less than 10%.

[0036] Multi-stage temperature control extraction Phase I: Mix all the medicinal materials in proportion, add 10 times the amount of deionized water, set the extraction temperature to 55°C, adjust the pH to 4.2 (add 0.3% citric acid), control the stirring speed at 120 rpm, extract for 1.8 hours, and filter the extract I.

[0037] The second stage: add 8 times the amount of 70% ethanol solution to the residue, raise the temperature to 70°C, extract for 2.5 hours, adjust the pH to 5.8 (add 0.2% sodium citrate buffer), stir at 150 rpm, and filter the extract II.

[0038] The third stage: add 6 times the amount of deionized water to the residue, raise the temperature to 90°C, extract for 1.2 hours, maintain the stirring speed at 180 rpm, and filter the extract III.

[0039] Concentration and purification The extracts I, II and III were combined and placed in a vacuum concentration device, and concentrated at 45°C and reduced pressure of 0.1 MPa to a total solid content of 65 mg / mL.

[0040] Use ultrafiltration membrane (molecular weight cutoff 1-5kDa) to remove impurities, and then use nanofiltration membrane (molecular weight cutoff 200-500Da) to enrich the effective ingredients to obtain a clarified concentrate.

[0041] Auxiliary material addition and molding Add 10 mg / portion of vitamin C pills and 3 mg / portion of vitamin E pills to the concentrate, stir evenly, and prepare drug powder using a spray drying device (temperature 80°C, spray pressure 0.3 MPa).

[0042] The dried medicinal powder and refined honey are mixed in a ratio of 1:0.4 to make water-honey pills. After the pills are formed, they are dried at 50°C until the moisture content is less than 8% to obtain the final product.

[0043] Example 2 Guishao Dihuang Pills with Improved Addition Ratio of Vitamin C and Vitamin E Processing of medicinal materials Weigh 38g of Angelica sinensis, 42g of white peony root with wine, 165g of Rehmannia root, 78g of Cornus officinalis with wine, 60g of Paeonia suffruticosa, 78g of Dioscorea opposita, 62g of Poria cocos, and 60g of Alisma orientalis.

[0044] After the medicinal materials are cleaned, the slices are 1mm thick and dried at 40°C for 9 hours to ensure that the moisture content of the medicinal materials is less than 8%.

[0045] Extraction process Stage 1: Add 12 times the amount of deionized water, set the extraction temperature to 50°C, the time to 1.5 hours, adjust the pH to 4.0 (add 0.2% citric acid solution), stir at 100 rpm, and filter the extract I.

[0046] The second stage: add 9 times the amount of 70% ethanol solution to the residue, adjust the temperature to 68°C, keep for 3 hours, adjust the pH to 6.2 (add 0.3% sodium citrate solution), and filter the extract II.

[0047] The third stage: add 7 times amount of pure water to the residue, set the temperature to 92°C, extract for 1 hour, stir at 200 rpm, and filter the extract III.

[0048] Concentration, purification and excipient addition The three-stage extracts were combined and concentrated at 50°C and reduced pressure of 0.08 MPa to a total solid content of 70 mg / mL.

[0049] The ultrafiltration and nanofiltration operations were the same as in Example 1. 15 mg / portion of vitamin C pills and 5 mg / portion of vitamin E pills (ratio 3:1) were added to the concentrated solution and stirred thoroughly.

[0050] Formulation After being treated by spray drying equipment, the drying temperature is 85℃ and the spray pressure is 0.4MPa to obtain the drug powder. The drug powder is mixed with refined honey in a ratio of 1:0.9 to make small honey pills. After forming and drying, the moisture content of the product is controlled at 6%.

[0051] Example 3 Guishao Dihuang Pills to improve the accuracy of extraction process Pretreatment of medicinal materials Weigh 40g of Angelica sinensis, 40g of white peony root with wine, 160g of Rehmannia glutinosa, 80g of Cornus officinalis with wine, 60g of Paeonia suffruticosa, 80g of Dioscorea opposita, 60g of Poria cocos, and 60g of Alisma orientalis.

[0052] The washed medicinal materials were sliced ​​into 0.8 mm thick slices and dried at 45°C for 10 hours. The moisture content of the medicinal materials was less than 10%.

[0053] Multi-stage temperature control extraction Stage 1: Place the medicinal materials in an extraction tank and add 11 times the amount of pure water. Set the extraction temperature to 52°C, adjust the pH to 4.5, extract for 1.7 hours, and filter the extract I.

[0054] The second stage: add 8 times the amount of 75% ethanol solution to the residue, extract at 70℃, extract for 2.2 hours, and filter the extract II.

[0055] The third stage: add 6.5 times the amount of deionized water to the residue, set the temperature to 89°C, extract for 1.3 hours, and filter the extract III.

[0056] Concentration and purification The extracts I, II and III were combined and concentrated at 48°C and the pressure was reduced to 0.09 MPa until the solid content was 80 mg / mL.

[0057] Impurities are removed by ultrafiltration membrane, and then key ingredients are enriched by nanofiltration membrane. The total active ingredient extraction rate of the concentrate is increased by 30% compared with traditional methods.

[0058] Molding process Add 20 mg / portion of vitamin C pills and 6 mg / portion of vitamin E pills (3.3:1 ratio) to the concentrate and stir evenly. Control the temperature of the spray drying equipment at 75°C and the spray pressure at 0.35 MPa. After preparing the powder, mix it with refined honey in a ratio of 1:0.6 to make water-honey pills and dry them to a moisture content of 7%.

[0059] Example 4 Guishao Dihuang Pills with improved stability and extended shelf life Medicinal material processing The weighing ratio of the medicinal materials is the same as that in Example 1, the slice thickness is 1 mm, the drying temperature is 40° C., and the drying time is 10 hours.

[0060] Multi-segment extraction The first stage extraction temperature was 55°C, the time was 2 hours, and the pH was adjusted to 4.3.

[0061] The second stage temperature was 70°C, the time was 2.8 hours, and the pH was 6.0.

[0062] The temperature of the third stage was 91°C and the time was 1.2 hours. The extracts from the three stages were combined and concentrated to a solid content of 65 mg / mL.

[0063] Addition of auxiliary materials Add 12 mg / portion of vitamin C pills and 4 mg / portion of vitamin E pills to the concentrate. The synergistic effect of the two can delay the oxidation of the active ingredients. The spray drying temperature is 80°C to obtain the powder, which is mixed with refined honey in a ratio of 1:0.5 to make honey pills. The moisture content of the product is controlled at 6%, and the shelf life is extended to 24 months after sealed packaging.

[0064] Comparative Example 1 (corresponding to Example 1, without using multi-stage temperature control extraction technology) Pretreatment of medicinal materials The types and proportions of medicinal materials were consistent with those in Example 1. After washing and slicing (thickness was 1 mm), they were dried at 45° C. for 10 hours.

[0065] Extraction process Add 12 times amount of pure water to the medicinal materials at one time, set the extraction temperature at 90°C for 3 hours without segmented temperature control, filter the extract and obtain a crude extract.

[0066] Concentration and purification The extract was directly concentrated (temperature 55°C, reduced pressure 0.1 MPa) to a total solid content of 65 mg / mL without ultrafiltration and nanofiltration separation steps, and the extract was directly used to prepare pills.

[0067] Auxiliary material addition and molding Add 10 mg / portion of vitamin C pills and 3 mg / portion of vitamin E pills to the concentrate, mix well and then spray dry (temperature 85°C), mix the obtained powder with refined honey in a ratio of 1:0.4 to make water-honey pills, and dry to obtain the finished product.

[0068] Comparative Example 2 (corresponding to Example 2, without the introduction of vitamin C and vitamin E) Pretreatment of medicinal materials The types, proportions, cleaning and slice thickness of the medicinal materials are consistent with those in Example 2, and the drying conditions are the same.

[0069] Extraction process The multi-stage temperature-controlled extraction process is consistent with Example 2, including three-stage temperature-controlled extraction: Stage 1: 50°C, 1.5 hours, pH adjusted to 4.0; Stage 2: 70°C, 3 hours, pH adjusted to 6.2; Stage 3: 92°C, 1 hour.

[0070] Concentration and purification The extracts were combined and concentrated in vacuo to a total solid content of 70 mg / mL. The ultrafiltration and nanofiltration separation processes were the same as in Example 2.

[0071] Molding process The spray-dried powder is directly mixed with refined honey in a ratio of 1:0.9 to prepare small honey pills without the addition of excipients such as vitamin C and vitamin E.

[0072] Comparative Example 3 (corresponding to Example 3, adjusting purification process parameters) Pretreatment of medicinal materials The types, proportions and slice thickness of the medicinal materials are consistent with those in Example 3, and the drying conditions are the same.

[0073] Extraction process The multi-stage temperature-controlled extraction process is consistent with Example 3, including three-stage extraction: Stage 1: 52°C, 1.7 hours, pH adjusted to 4.5; Stage 2: 70°C, 2.2 hours; Stage 3: 89°C, 1.3 hours.

[0074] Concentration and purification The extracts were combined and concentrated in vacuo to a solid content of 80 mg / mL. In the purification process, only the traditional coarse filtration method was used without using ultrafiltration and nanofiltration technology. The impurities in the extract were not further removed and directly entered the next step of the molding process.

[0075] Auxiliary material addition and molding Add 20 mg / portion of vitamin C pills and 6 mg / portion of vitamin E pills to the concentrate, mix well and spray-dry. Mix the powder with refined honey in a ratio of 1:0.6 to make water-honey pills.

[0076] Comparative Example 4 (corresponding to Example 4, adjusting the refining ratio and molding parameters) Pretreatment of medicinal materials The types, proportions and slice thickness of the medicinal materials are consistent with those in Example 4, and the drying conditions are the same.

[0077] Extraction process The multi-stage temperature-controlled extraction process is consistent with that in Example 4: Stage 1: 55°C, 2 hours; Stage 2: 70°C, 2.8 hours; Stage 3: 91°C, 1.2 hours.

[0078] Concentration and purification The operations of combining the extracts, vacuum concentration, ultrafiltration and nanofiltration were consistent with those in Example 4. The total solid content was 65 mg / mL.

[0079] Auxiliary material addition and molding Vitamin C 12mg / portion pills and vitamin E 4mg / portion pills were added to the concentrate. The spray-dried powder was mixed with refined honey at a ratio of 1:0.2. The pill-making process failed to fully form the pills, the surface of some pills was rough, and the moisture content of the finished product was uneven after drying.

[0080] Comparative Example 5 (corresponding to Example 1, changing the spray drying process parameters) Pretreatment of medicinal materials The types, proportions and slice thickness of the medicinal materials are consistent with those in Example 1, and the drying conditions are the same.

[0081] Extraction process The multi-stage temperature-controlled extraction process is consistent with Example 1, including three-stage extraction temperature and time settings.

[0082] Concentration and purification The extracts were combined and concentrated in vacuo to a total solid content of 65 mg / mL. The ultrafiltration and nanofiltration separation steps were the same as in Example 1.

[0083] Auxiliary material addition and molding Vitamin C 10 mg / portion pills and vitamin E 3 mg / portion pills were added to the concentrate, and the spray drying process parameters were adjusted to: drying temperature 95°C, spray pressure 0.5 MPa. After drying, the powder particles were too fine and some powders agglomerated, affecting the molding. The powder and refined honey were mixed in a ratio of 1:0.4, the surface viscosity of the water-honey pills increased, and drying was incomplete.

[0084] Experiment 1: Comparison of active ingredient extraction rate Objective: To verify whether the multi-stage temperature-controlled extraction process of the present invention can improve the extraction rate of active ingredients (such as catalpol, paeoniflorin, and polysaccharides).

[0085] Experimental methods: Sample preparation The extracts were prepared using Example 1, Example 3, Comparative Example 1 and Comparative Example 3, and the three-stage extraction (or the corresponding one-time extraction) was completed using the respective extraction processes.

[0086] The extracts were combined and concentrated to a total solid content of 60 mg / mL.

[0087] Test indicators Determination of catalpol content: High performance liquid chromatography (HPLC) was used, the chromatographic column was C18, the mobile phase was methanol-water (50:50, v / v), and the detection wavelength was 280 nm.

[0088] Determination of paeoniflorin content: HPLC was used with a C18 column, acetonitrile-water (30:70, v / v) as the mobile phase, and the detection wavelength was 230 nm.

[0089] Determination of polysaccharide content: Use the phenol-sulfuric acid method, the detection wavelength is 490nm, and the content is calculated based on the glucose standard curve.

[0090] Experimental setup Experimental groups: Example 1 and Example 3.

[0091] Control group: Comparative Example 1 and Comparative Example 3.

[0092] Each group was tested 3 times and the average value was taken.

[0093] Experiment 2: Comparison of formulation stability Objective: To evaluate the protective effect of the present invention on the active ingredients in the preparation after adding vitamin C and vitamin E, and to verify the stability of the product during storage.

[0094] Experimental methods: Sample preparation Small honey pills were prepared using Example 2 (containing vitamins C and E) and Comparative Example 2 (excluding vitamins C and E), respectively, and stored after being formed according to their respective processes.

[0095] Experimental setup Under accelerated aging conditions of 40°C and 75% relative humidity, the active ingredient content and oxidative degradation of the samples were tested at 0 days, 7 days, 14 days, 30 days, and 60 days.

[0096] Test indicators Degradation rate of catalpol and paeoniflorin: HPLC was used to determine the content of active ingredients at each storage time point, and the degradation rate was calculated by comparing with the initial value.

[0097] Antioxidant capacity determination: The antioxidant properties of the product were evaluated by DPPH free radical scavenging capacity test, with a detection wavelength of 517nm.

[0098] Moisture content: Karl Fischer titration was used to detect the changes in moisture content in the pills.

[0099] Experiment 3: Comparison of extract purity Objective: To verify whether the ultrafiltration and nanofiltration technologies in the present invention can significantly improve the purity of the extract and the enrichment efficiency of key active ingredients.

[0100] Experimental methods: Sample preparation Example 1 and Comparative Example 1 were used for comparison, and the extract was purified by using an ultrafiltration-nanofiltration separation process and a traditional coarse filtration method, respectively.

[0101] The two groups of extracts were concentrated to 60 mg / mL and then analyzed.

[0102] Experimental setup The comparative experiment was set up into two groups: the extract of Example 1 (ultrafiltration-nanofiltration separation) and the extract of Comparative Example 1 (conventional coarse filtration).

[0103] Test indicators Total impurity content: The ratio of macromolecules and impurities in the residue (1 kDa fraction) was determined gravimetrically.

[0104] Catalpol and paeoniflorin content: HPLC was used to determine the enrichment of the two components in the extract.

[0105] Color and clarity: The clarity of the extracts was compared by optical density measurement (wavelength 450nm).

[0106] Experiment 4: Comparison of immune regulation ability Objective: To verify whether the preparation of the present invention has a better regulating effect on the immune function of mice than the comparative example.

[0107] Experimental methods: Animal experiment design 48 mice were selected and randomly divided into the following groups, 8 mice in each group: Example 1 group (multi-stage temperature control + vitamin C and E); Example 2 group (multi-stage temperature control + vitamins C and E); Comparative Example 1 (single high temperature extraction, without vitamin C and E); Comparative Example 2 (multi-stage temperature-controlled extraction, without vitamin C and E); Blank control group (no treatment).

[0108] Experimental setup The mice were intragastrically administered with the corresponding preparations of the above groups every day (at a dose of 0.2 g / kg body weight) for 14 consecutive days.

[0109] Immune indexes were tested on the 15th day.

[0110] Test indicators Thymus and spleen index: Take the thymus and spleen of mice and calculate their weight / body weight (mg / g).

[0111] Serum IL-2 and IFN-γ levels: Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of immune-related factors in serum.

[0112] T lymphocyte proliferation rate: The MTT method was used to detect the proliferation ability of mouse spleen cells under ConA stimulation.

[0113] Experiment 1: Comparative experiment on active ingredient extraction rate Purpose The effect of multi-stage temperature-controlled extraction technology on the extraction efficiency of active ingredients such as catalpol, paeoniflorin, and polysaccharides was verified, and the necessity and scientificity of extraction process optimization were demonstrated by comparing the examples with the comparative examples.

[0114] Sample preparation Example 1 and Example 3: According to the multi-stage temperature-controlled extraction process, three stages of extraction were completed respectively: the first stage was low-temperature extraction (50-55°C), the second stage was medium-temperature extraction (65-75°C), and the third stage was high-temperature extraction (85-95°C). The extracts were combined and concentrated to a solid content of 60 mg / mL.

[0115] Comparative Example 1: Single high temperature extraction (90°C) was used, the extraction time was 3 hours, and there was no segmented operation. The extract was concentrated to 60 mg / mL.

[0116] Comparative Example 3: A three-stage temperature-controlled extraction was adopted, but ultrafiltration and nanofiltration were not performed, and the extract was directly coarsely filtered and concentrated to 60 mg / mL.

[0117] Sample testing Determination of catalpol content: HPLC method was used, C18 chromatographic column, the mobile phase was methanol-water (50:50, v / v), and the detection wavelength was 280nm.

[0118] Determination of paeoniflorin content: HPLC method was used, C18 chromatographic column, the mobile phase was acetonitrile-water (30:70, v / v), and the detection wavelength was 230 nm.

[0119] Determination of polysaccharide content: Phenol-sulfuric acid method was used, phenol solution and sulfuric acid were added to develop color, and the absorbance was measured at a wavelength of 490 nm.

[0120] Each sample was measured three times in parallel and the average value was taken.

[0121] Experimental data Table 1 Comparison of active ingredient extraction rates between examples and comparative examples Experimental Summary This experiment reveals the significant advantages of multi-stage temperature-controlled extraction technology in extracting active ingredients. From the data, the extraction efficiency of catalpol and paeoniflorin in the examples is generally high, while the decomposition of catalpol and the loss of paeoniflorin in Comparative Example 1 are particularly obvious due to the use of a single high-temperature extraction process. This also shows that thermosensitive components require precise temperature control, and a single extraction technology is difficult to take into account the stability and release of multiple components. This result proves the scientific nature of the multi-stage temperature-controlled extraction scheme, and more thermosensitive components are retained through temperature control and pH adjustment.

[0122] From the extraction results of polysaccharide content, the embodiment also has obvious advantages over comparative example 3. Although comparative example 3 has a slight improvement in the extraction of certain components, the lack of purification steps leads to more impurities, which ultimately affects the concentration and quality of polysaccharides. In the embodiment, the use of ultrafiltration and nanofiltration effectively reduces the interference of macromolecular impurities and improves the enrichment effect of polysaccharides. This combined process further improves the purity of the extract through separation and enrichment, laying a foundation for the efficacy of the drug.

[0123] This is not just a comparison of numbers, but a logical verification. High-temperature extraction is simple, but the damage to the medicinal ingredients is also obvious. The multi-stage extraction technology adopted in the embodiment not only avoids the degradation of heat-sensitive components, but also provides more suitable extraction conditions for macromolecular components. This design concept not only solves the shortcomings of traditional processes, but also provides a modern and scientific solution for the efficient utilization of active ingredients.

[0124] Experiment 2: Comparative experiment on the stability of preparations Purpose By conducting accelerated aging tests on the pills prepared in Example 2 and Comparative Example 2, the protective effects of vitamins C and E on the active ingredients in the preparations were evaluated, and the stability improvement effect of the preparations of the present invention was verified.

[0125] Sample preparation Example 2: Guishao Dihuang Pills containing vitamin C (15 mg / pill) and vitamin E (5 mg / pill) were prepared, and small honey pills were made according to the process, and the moisture content was controlled at 6%.

[0126] Comparative Example 2: Guishao Dihuang Pills were prepared according to the comparative example process, without adding vitamin C and E, and the remaining steps were the same.

[0127] Aging test The samples were stored in an accelerated aging chamber at 40°C and 75% relative humidity, and the sampling times were 0 days, 7 days, 14 days, 30 days, and 60 days.

[0128] The contents of catalpol and paeoniflorin were detected at each time point, and the antioxidant capacity (DPPH free radical scavenging rate) was measured. Each group of samples was measured in parallel 3 times.

[0129] Test Method Determination of catalpol and paeoniflorin content: The same as experiment 1, determined by HPLC.

[0130] DPPH free radical scavenging rate: After adding 0.1 mM DPPH solution, the absorbance was measured at a wavelength of 517 nm and the scavenging rate was calculated.

[0131] Determination of moisture content: Karl Fischer titration method was used, 2 g of sample was taken each time for determination, and the average value was calculated.

[0132] Experimental data Time (days) Sample No. Catalpol content (mg / mL) Paeoniflorin content (mg / mL) DPPH removal rate (%) Moisture content (%) 0 Example 2 2.36 1.92 86.5 6.1 Comparative Example 2 2.31 1.88 85.8 6.0 7 Example 2 2.30 1.87 83.4 6.3 Comparative Example 2 2.19 1.76 78.2 6.2 14 Example 2 2.22 1.78 81.2 6.4 Comparative Example 2 2.06 1.62 72.9 6.5 30 Example 2 2.10 1.62 76.5 6.5 Comparative Example 2 1.82 1.34 65.1 6.7 60 Example 2 1.95 1.48 71.8 6.7 Comparative Example 2 1.60 1.10 56.2 6.9 Experimental Summary The protective effects of vitamins C and E in the preparation are very intuitive. Under accelerated aging conditions, the decrease in the content of catalpol and paeoniflorin in the example is much lower than that in the control example, especially after storage for 30 days, the example still maintains a relatively high content, while the control example has begun to decline significantly. The key here is that vitamin C provides a rapid antioxidant barrier for the active ingredients in the aqueous phase, while vitamin E stabilizes the overall oxidative environment inside the pills through its fat-soluble properties. The combination of the two makes the entire system perform better under oxidative stress.

[0133] The DPPH free radical scavenging rate further supports this point. In the examples, the rate of decline of antioxidant capacity slowed down significantly, while the clearance rate of the comparative example dropped significantly after 14 days. For Chinese medicine pills, antioxidant performance is directly related to the maintenance of drug efficacy. The comparative example gradually lost its effectiveness during storage, which obviously cannot meet the needs of modern consumers for efficient and stable drugs. The examples solve this core problem by introducing functional excipients.

[0134] The change in moisture content is also very convincing. The moisture control of the embodiment is more stable, while the moisture content of the comparative example fluctuates slightly in the later stage due to the lack of sufficient protective auxiliary materials. This fluctuation may further accelerate the degradation of the ingredients. It is the optimization of these details that reflects the advantages of the present invention in stability. The experimental results not only illustrate the synergistic effect of vitamins C and E, but also clarify the scientificity and reliability of the formulation design of the present invention.

[0135] Experiment 3: Comparative experiment on immune regulation ability Purpose The effects of Guishao Dihuang Pills prepared in the embodiment and the comparative example in enhancing immune function were compared through animal experiments, with a focus on evaluating the synergistic effect of vitamins C and E and the effects of multi-stage temperature-controlled extraction technology on immune regulation, thereby verifying the contribution of the present invention to improving immunity.

[0136] Experimental procedures Animal grouping and treatment The experiment used 8-week-old healthy Kunming mice (♂), weighing about 20±2g, a total of 48 mice, and randomly divided them into the following 6 groups, 8 mice in each group: Blank control group: The mice were given an equal amount of normal saline by intragastric administration of 0.2 mL per day.

[0137] Group 1 of Example 1 was intragastrically administered with the Guishao Dihuang Pill solution (0.2 g / kg) prepared in Example 1 every day.

[0138] Group 2 of Example 2 was intragastrically administered with the Guishao Dihuang Pill solution (0.2 g / kg, containing vitamins C and E) prepared in Example 2 every day.

[0139] Comparative Example 1 group: The Guishao Dihuang Pill solution (0.2 g / kg, single high-temperature extraction) prepared in Comparative Example 1 was intragastrically administered every day.

[0140] Comparative Example 2 group: The Guishao Dihuang Pill solution (0.2 g / kg, without vitamin C and E) prepared in Comparative Example 2 was intragastrically administered every day.

[0141] Comparative Example 3 group: The Guishao Dihuang Pill solution prepared in Comparative Example 3 (0.2 g / kg, without purification step) was intragastrically administered every day.

[0142] Experimental cycle After 14 days of continuous gavage, the mice were fasted for 12 hours and samples were collected to detect immune-related indicators.

[0143] Experimental testing Spleen index: The spleen of mice was removed, weighed and spleen weight / body weight (mg / g) was calculated.

[0144] Thymus index: remove the thymus, weigh it and calculate the thymus weight / body weight (mg / g).

[0145] Serum IL-2 and IFN-γ levels: Enzyme-linked immunosorbent assay (ELISA) was used to detect the concentrations of IL-2 and IFN-γ in serum (pg / mL).

[0146] T lymphocyte proliferation rate: Mouse spleen cells were taken and stimulated by adding ConA (final concentration 5 μg / mL). The proliferation capacity was determined by MTT method, and the OD value reflected the proliferation rate.

[0147] Experimental data Table 3. Comparison of immunomodulatory ability between examples and comparative examples Experimental Summary This experiment further verified the immunomodulatory advantage of the present invention through an animal model. The spleen index and thymus index of the mice added with vitamins C and E in Example 2 were significantly higher than those of the control group and the blank control group, which shows that the excipients have a positive effect on the development and functional enhancement of immune organs, especially in the improvement of the spleen index. The performance of the embodiment is particularly outstanding, which also confirms that the combination of multi-stage temperature control extraction technology and excipient design can maximize the retention and utilization of the active ingredients of traditional Chinese medicine, while the control group of single high-temperature extraction cannot achieve such an effect.

[0148] From the detection data of IL-2 and IFN-γ, the serum cytokine level of Example 2 is significantly better than that of Example 1, and further higher than all comparative examples, which shows that the synergistic effect of vitamins C and E is not only antioxidant, but also can enhance the overall effect by regulating the immune pathway. The high level of IFN-γ also suggests that the improved pill has a strong stimulating effect on cellular immunity, which is consistent with the proliferation results of T cells during antigen activation. Among the comparative examples, especially Comparative Example 2 where no functional excipients are introduced, the immune indicators are significantly insufficient compared with the examples, indicating that the prior art cannot solve the loss of active ingredients in extraction and stability.

[0149] Finally, the results of T cell proliferation rate are also very interesting. The stimulation effect of Example 2 is optimal, reflecting the potential promotion of vitamin excipients on immune cell activity, while the cell culture environment of Comparative Example 3 is limited due to the large number of impurities in the extract, and the proliferation rate is much lower than that of the embodiment. This comparison highlights the impact of the extraction and purification process on the actual effect of the product. The present invention solves the shortcomings of traditional Guishao Dihuang Pills in improving immune function from a mechanism perspective through reasonable process optimization and excipient design. This is not only a technical improvement, but also a complete application innovation.

[0150] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A Guishao Dihuang Pill, characterized in that: The composition comprises the following components in parts by weight: Angelica: 35-45 parts; White peony root with wine: 35-45 parts; Rehmannia root: 150-170 parts; Wine-boiled cornus: 75-85 portions; Peony bark: 55-65 parts; Chinese Yam: 75-85 servings; Poria: 55-65 parts; Alisma orientalis: 55-65 parts; Each pill contains 5-20 mg of vitamin C and 2-10 mg of vitamin E, and the pills are prepared by the following process: The method includes the following steps: pretreatment of medicinal materials: cleaning, slicing and drying of medicinal materials; multi-stage temperature-controlled extraction of active ingredients; concentration and purification of extracts; addition and mixing of auxiliary materials; Formulation of preparations.

2. The Guishao Dihuang Pill according to claim 1, characterized in that: The added ratio of vitamin C and vitamin E is 3:

1.

3. The Guishao Dihuang Pill according to claim 1, characterized in that: The pills are water-honey pills, which are prepared by adding 35-50g of refined honey to every 100g of medicinal material powder.

4. The Guishao Dihuang Pill according to claim 1, characterized in that: The pills are small honey pills, which are prepared by adding 80-110g refined honey to every 100g medicinal material powder.

5. A method for preparing Guishao Dihuang Pills, applied to the Guishao Dihuang Pills according to any one of claims 1 to 4, characterized in that: The following steps are involved: Wash the herbs and slice them into slices, the thickness of slices of Radix Rehmanniae glutinosae is 0.5-2.0 mm, and the thickness of slices of other herbs is 1.0-2.0 mm; The multi-stage temperature-controlled extraction was carried out under the following conditions: the temperature of the first stage was 50-55°C, the extraction time was 1.5-2 hours, and the pH was adjusted to 4.0-4.5; the temperature of the second stage was 65-75°C, the extraction time was 2-3 hours, and the pH was adjusted to 5.5-6.5; the temperature of the third stage was 85-95°C, and the extraction time was 1-1.5 hours; The extracts are combined and concentrated to a solid content of 60 to 80 mg / mL, and purified by ultrafiltration and nanofiltration membranes; Vitamin C and vitamin E are added, spray-dried, prepared into powder, and mixed with refined honey to form pills.

6. The method for preparing Guishao Dihuang Pills according to claim 5, characterized in that: The thickness of the medicinal material slices is 0.5-1.0 mm, the drying temperature is 40-50° C., and the drying time is 8-12 hours.

7. The method for preparing Guishao Dihuang Pills according to claim 5, characterized in that: During the first stage of extraction, the weight-to-volume ratio of the medicinal material to water is 1:10 to 1:

12.

8. The method for preparing Guishao Dihuang Pills according to claim 5, characterized in that: The concentration step is carried out at a reduced pressure of 0.08-0.1 MPa and a concentration temperature of 45-55°C.

9. The method for preparing Guishao Dihuang Pills according to claim 5, characterized in that: The temperature range of the spray drying is 70-90° C., and the spray pressure is 0.2-0.5 MPa.

10. The method for preparing Guishao Dihuang Pills according to claim 5, characterized in that: The added ratio of the refined honey during the pill making process is 35 to 110 g per 100 g of medicinal material powder.

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