Preparation method of radix bupleuri and radix paeoniae alba oral liquid

Abstract

The invention discloses a preparation method of radix bupleuri and radix paeoniae alba oral liquid, and belongs to the technical field of traditional Chinese medicine preparations and pharmaceutical engineering.The method comprises the steps that according to different physicochemical properties of radix bupleuri, radix paeoniae alba, fructus aurantii immaturus and liquorice, a differential airflow superfine grinding technology is adopted, hard medicinal materials are ground till D90 is smaller than or equal to 15 microns, and crisp medicinal materials are ground till D90 is smaller than or equal to 25 microns; then gradient temperature immersion and variable-temperature countercurrent dynamic extraction are carried out, variable-frequency ultrasonic assistance (25-35 kHz) is introduced in the high-temperature extraction stage, and dissolution of effective components is improved through the synergistic effect; carrying out graded filtration on the extracting solution through 0.2 mu m and 5000Da ceramic membranes, so as to effectively remove impurities and retain effective parts of small molecules; finally, three-effect falling film low-temperature vacuum concentration and xylitol-stevioside in a specific proportion are adopted for taste correction. The preparation method solves the problems of low extraction rate, more impurities and poor stability of a traditional process, and the prepared oral liquid is high in active ingredient content, good in clarity, good in taste, good in process reproducibility and suitable for industrial production.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine preparations and pharmaceutical engineering technology, and in particular to a method for preparing Chai Shao oral liquid. Background Technology

[0002] Chai Shao Oral Liquid originates from a modified version of the classic formula "Si Ni San," primarily composed of Bupleurum, White Peony Root, Fructus Aurantii Immaturus, and Glycyrrhizae Radix et Rhizoma. It possesses the effects of soothing the liver and regulating Qi, relieving liver pain, and is widely used to treat symptoms such as liver and gallbladder Qi stagnation and hypochondriac distending pain. However, the preparation of traditional Chinese medicine oral liquids often employs a simple water decoction method, which presents the following technical bottlenecks: 1. Low extraction efficiency: Traditional decoction methods rely on natural penetration and diffusion. For hard-textured ingredients such as Bupleurum and Citrus aurantium, it is difficult to completely dissolve the effective components (such as saikosaponins and synephrine); for heat-sensitive components, prolonged high-temperature decoction can easily lead to decomposition.

[0003] 2. Difficulty in removing impurities: Conventional water extracts contain a large amount of tannins, resins, polysaccharides and proteins, which not only affect the taste, but also easily produce precipitation or turbidity during storage, affecting the clarity and stability of the preparation.

[0004] 3. Crude process: Existing technologies mostly adopt the "one-pot cooking" mode, which lacks differentiated treatment for medicinal materials with different properties, resulting in significant loss of effective ingredients and obvious batch-to-batch differences.

[0005] Therefore, developing a highly efficient preparation method that can differentiate the physicochemical properties of different medicinal materials in Chai Shao oral liquid and combine it with modern separation technology is a technical problem that urgently needs to be solved. Summary of the Invention

[0006] The purpose of this invention is to provide a method for preparing Chai Shao oral liquid. This method solves the problems of incomplete extraction of effective ingredients, incomplete removal of impurities, and poor formulation stability by implementing a refined process of "differentiated ultrafine pulverization + gradient temperature soaking + variable temperature countercurrent dynamic extraction + ultrasonic assistance + ceramic membrane fine filtration + low temperature concentration". This method achieves efficient and stable production of Chai Shao oral liquid.

[0007] To achieve the above-mentioned objectives, the technical solution adopted by this invention is as follows: A method for preparing a Chai Shao oral liquid, characterized by comprising the following steps: (1) Pre-processing of medicinal materials and control of processed products This invention selects authentic medicinal materials and emphasizes the influence of processing on efficacy. Processing Bupleurum with vinegar enhances its liver-soothing and pain-relieving effects, and vinegar promotes the dissolution of saponins; stir-frying Paeonia lactiflora with wine mitigates its cold nature and enhances its liver-soothing effect. Specific process: Bupleurum slices are mixed with rice vinegar, moistened, and then stir-fried over low heat until dry (15kg / 100kg vinegar); Paeonia lactiflora slices are mixed with rice wine, moistened, and then stir-fried over low heat until slightly yellow (10kg / 100kg wine). Honey-processing of Glycyrrhiza uralensis and wheat bran-frying of Citrus aurantium can also be performed as needed, but this embodiment uses raw products to preserve alkaloid activity.

[0008] (2) Differentiated ultrafine grinding (airflow grinding) Traditional ball mills generate a lot of heat during grinding, which can easily lead to the loss of heat-sensitive components. This invention uses fluidized bed airflow grinding.

[0009] Bupleurum and Citrus aurantium: These have a hard texture and many fiber bundles. The grinding pressure was set at 0.7 MPa, the feed rate at 12 kg / h, and the target particle size D90 = 12 ± 3 μm. At this particle size, the cell wall breakage rate was >95%, and the instantaneous dissolution rate of saikosaponins increased by 3 times.

[0010] White peony root and licorice root: Contain starch and sugars, easily causing adhesion. Set the grinding pressure to 0.6 MPa, with a target particle size D90 = 20 ± 5 μm. Avoid excessive grinding, which could lead to increased hygroscopicity and adsorption of active ingredients.

[0011] Key point: The two powders are pulverized separately and then mixed to avoid "segregation" caused by differences in hardness and to ensure uniform mixing.

[0012] (3) Gradient temperature soaking and variable temperature countercurrent extraction Gradient temperature soaking: First, the medicinal material is moistened at a low temperature (30℃) to prevent surface protein coagulation; then, the temperature is increased (50℃) to soften the tissue and prepare for extraction. This process can shorten the actual extraction time by 30%.

[0013] Variable temperature countercurrent: The extraction tank is designed with a top inlet and bottom outlet (or circulation) structure.

[0014] First stage (78℃, 50min): Extraction of paeoniflorin under mild conditions (highly heat-sensitive).

[0015] Second stage (92℃, 35min): Extraction of saikosaponins (with strong heat resistance) and glycyrrhizic acid under strong conditions.

[0016] Dynamic circulation: The pump circulates the liquid medicine at a flow rate of 1.8 m / s, creating turbulence, which breaks the diffusion boundary layer on the surface of the medicinal material and maximizes the concentration difference.

[0017] (4) Ultrasonic field strength assistance (not simple superposition) The ultrasound was not activated continuously, but only during the second high-temperature phase. The frequency was set to 28kHz (targeting the lowest cavitation threshold of saikosaponins), and the sound intensity was 0.8W / cm². 2 The mechanical effect of ultrasound disrupts residual cell walls, while the thermal effect assists molecular motion. This synergistic effect, combined with high-temperature extraction, increases the total extraction rate by 12%–15%.

[0018] (5) Ceramic membrane microfiltration for impurity removal Traditional alcohol precipitation methods are time-consuming and consume large amounts of alcohol. This invention uses a 0.2μm / 5000Da dual-stage ceramic membrane.

[0019] 0.2μm membrane: traps bacteria, fibers, and starch granules, ensuring sterility and clarity.

[0020] 5000Da ultrafiltration membrane: retains large molecular proteins, tannins, and polysaccharides (allergens and turbidity sources), while the permeate contains small molecular effective components (saponins, flavonoids, alkaloids).

[0021] This step can replace alcohol precipitation, reducing ethanol usage by 80% and increasing the recovery rate of active ingredients by 20%.

[0022] (6) Low-temperature vacuum concentration A triple-effect falling film evaporation system is employed. The first effect utilizes the secondary steam from the second effect as a heat source, resulting in energy efficiency. The temperature of the third effect is strictly controlled to ≤50℃, and the vacuum degree is ≥-0.09MPa. Compared to atmospheric pressure concentration, the degradation rate of paeoniflorin is reduced from 8% to below 1%.

[0023] (7) Precipitation and flavoring Although membrane separation is performed, a small amount of alcohol precipitation is still needed to remove trace amounts of polar impurities. The alcohol content is controlled at 65%, at which point acidic components such as glycyrrhizic acid are less likely to precipitate. Xylitol (for preventing tooth decay and suitable for diabetic patients) combined with steviol glycosides is used as a flavoring agent to mask the strong bitterness of Bupleurum chinense.

[0024] As an improvement, the beneficial effects of the present invention are as follows: 1. Significantly improved extraction rate: The transfer rate of saikosaponins a+d is ≥85% (approximately 60% using the traditional method), and the transfer rate of paeoniflorin is ≥90%.

[0025] 2. Excellent clarity and stability: After filtration through the ceramic membrane, the light transmittance of the liquid is ≥98%, and there is no sedimentation or flocculation after 12 months of storage.

[0026] 3. Shortened production cycle: Dynamic extraction combined with ultrasound reduces the total extraction time from the traditional 4 hours to 1.5 hours.

[0027] 4. Energy saving and environmental protection: Membrane separation replaces a large amount of alcohol precipitation, reducing the cost of ethanol recovery and reducing the COD value of wastewater by 70%.

[0028] 5. Pharmacodynamic advantages: Animal experiments have shown that the equivalent dose of this product in terms of analgesia and anti-inflammation is only 1 / 2 of that of the traditional process. Detailed Implementation

[0029] To make the content of this invention easier to understand, the technical solutions in the embodiments of this invention will be clearly and completely described below in conjunction with the embodiments of this invention.

[0030] Example 1: Standard process preparation (optimal mode) 1. Ingredients: 250g vinegar-processed Bupleurum chinense, 250g wine-processed Paeonia lactiflora, 250g wheat bran-fried Citrus aurantium, and 250g roasted Glycyrrhiza uralensis.

[0031] 2. Grinding: Vinegar-processed Bupleurum chinense and wheat bran-fried Citrus aurantium: airflow pulverization, D90=13μm.

[0032] White peony root and prepared licorice root: airflow pulverization, D90=22μm.

[0033] 3. Warm soaking: Add 6 times the amount of water to the mixed powder, stir at 30℃ for 20 minutes, then raise the temperature to 50℃ and stir for 10 minutes.

[0034] 4. Extraction: Add 8 times the amount of water and extract at 78℃ for 50 minutes (stirring at 70 rpm); raise the temperature to 92℃ and simultaneously turn on 28kHz ultrasound (0.8W / cm). 2 Extraction time was 35 minutes. The entire process was dynamically circulated at a flow rate of 1.8 m / s.

[0035] 5. Filtration: The extract is microfiltered through a 0.2μm alumina ceramic membrane and then filtered through a 5000Da ultrafiltration membrane.

[0036] 6. Concentration: Triple-effect falling film concentration, triple-effect temperature 50℃, vacuum -0.092MPa, concentration to a relative density of 1.12.

[0037] 7. Alcohol precipitation: Add ethanol to 65%, refrigerate for 18 hours, and collect the supernatant to recover the ethanol.

[0038] 8. Flavoring and filling: Add 30g xylitol and 3g steviol glycosides, adjust pH to 5.0, add water to 1000ml, filter with 0.22μm filter, fill, and sterilize at 115℃ for 30min.

[0039] Test results: Saikosaponin a+d content 13.2 mg / 100ml, paeoniflorin 28.5 mg / 100ml. Transmittance 99.2%. Accelerated test (40℃ / 75%RH) for 6 months, content decreased by 4.5%.

[0040] Example 2: Comparison of Optimized Particle Size Group A (this invention): Bupleurum / Fructus Aurantii Immaturus D90=13μm; Paeoniae Radix Alba / Radix Glycyrrhizae Radix et Rhizoma D90=22μm.

[0041] Group B (Comparative Example 1): All medicinal materials were uniformly pulverized to D90=40μm (conventional fine powder).

[0042] Group C (Comparative Example 2): All medicinal materials were uniformly pulverized to D90=10μm (over-pulverization).

[0043] result: The extract from group A was clear, without gelatinization, and had the highest total saponin content.

[0044] Group B showed incomplete extraction; the residue had a hard core when squeezed by hand.

[0045] Group C experienced severe heat generation during the pulverization process (up to 80℃), resulting in a small amount of degradation of paeoniflorin and clogging of the filter membrane during filtration (due to high viscosity).

[0046] Conclusion: Differentiated particle size control is key to achieving efficient extraction and smooth filtration.

[0047] Example 3: Synergistic effect of extraction temperature and ultrasound Experimental Design: 1#: Room temperature + no ultrasound (traditional impregnation).

[0048] 2#: Temperature variation of 78℃ to 92℃, no ultrasound.

[0049] 3#: Temperature variation of 78℃ to 92℃, ultrasonic throughout (28kHz).

[0050] 4#: Temperature variation of 78℃ to 92℃, ultrasound only at the 92℃ stage (this invention).

[0051] result: Extraction rate of #1 was only 45%, and it took 4 hours.

[0052] Extraction rate of #2 was 78%, taking 1.5 hours.

[0053] The extraction rate of #3 is 82%, but energy consumption increases and some heat-sensitive impurities dissolve more.

[0054] #4 has an extraction rate of 86%, moderate energy consumption, and few impurities.

[0055] Conclusion: Staged ultrasound-assisted treatment (only during the high-temperature period) can maximize the synergistic effect and avoid ineffective energy consumption and impurity dissolution.

[0056] Example 4: Comparison of Filtration Methods Experimental Design: A: Plate and frame filter + diatomaceous earth (traditional).

[0057] B: Alcohol precipitation (65% alcohol content).

[0058] C: 0.2μm+5000Da ceramic membrane (this invention).

[0059] result: A: The clarity is acceptable, but the diatomaceous earth adsorbs some of the effective components (loss of about 8%), and there is a risk of introducing foreign matter.

[0060] B: The clarity is good, but 15% of the glycyrrhizic acid precipitation is lost, and it takes 24 hours.

[0061] C: Excellent clarity (microporous filter membrane grade), no adsorption loss of active ingredients, and operation time <1 hour.

[0062] Conclusion: Ceramic membrane filtration has an overwhelming advantage in retaining active ingredients and improving efficiency.

[0063] Example 5: Validation of the concentration process Experimental Design: A: Atmospheric pressure concentration (100℃).

[0064] B: Single-effect vacuum concentration (60℃).

[0065] C: Triple-effect falling film concentration (50°C, this invention).

[0066] result: A: The concentrated paste is dark black in color, has a burnt smell, and paeoniflorin is undetectable (it has completely decomposed).

[0067] B: Darker in color, with 85% retention of paeoniflorin.

[0068] C: Light yellow in color, with a 98% retention rate of paeoniflorin, and energy consumption is only 40% of that of A.

[0069] Conclusion: Low-temperature vacuum concentration is a necessary condition for ensuring the survival of heat-sensitive components.

[0070] Example 6: The Influence of Different Processing Methods Experimental Design: Sample 1: Raw Bupleurum chinense and raw White Peony Root.

[0071] Sample 2: Vinegar-processed Bupleurum chinense and wine-processed Paeonia lactiflora (this invention).

[0072] Sample 3: Turtle blood Bupleurum, stir-fried white peony root.

[0073] result: Sample 1: It has a high content of volatile oils, but its liver-soothing and pain-relieving effects are weak, and it is highly irritating to the stomach.

[0074] Sample 2: The total saponin content decreased slightly (due to dilution by vinegar / alcohol), but the saikosaponins were more easily dissolved after vinegar treatment, and the bioavailability increased by 30%.

[0075] Sample 3: The process is complex and the cost is high, but the efficacy is not significantly different.

[0076] Conclusion: Vinegar-processed Bupleurum and wine-fried White Peony Root is the most cost-effective processing combination.

[0077] Example 7: Screening of Flavoring Agents Experimental Design: Option 1: Sucrose (5%).

[0078] Option 2: Aspartame (0.1%).

[0079] Option 3: Xylitol (3%) + Stevioside (0.3%) (This invention).

[0080] result: Option 1: The sweetness is strong, and the effect of masking bitterness is not great. Moreover, the high sugar content is not good for patients (especially those with diabetes or liver and gallbladder diseases).

[0081] Option 2: It has a bitter aftertaste and an unnatural taste.

[0082] Option 3: It has a refreshing taste, completely masks the bitterness, leaves no aftertaste, and has auxiliary effects in protecting the liver and lowering blood sugar.

[0083] Conclusion: The combination of xylitol and steviol glycosides in a specific ratio not only improves the taste but also increases the added value of the formulation.

[0084] Example 8: Optimization of Dynamic Circulation Rate Extraction experiments were conducted with different circulation flow rates (0.5, 1.0, 1.5, 2.0, 2.5 m / s).

[0085] Results: When the flow rate is <1.0 m / s, the extraction rate increases significantly with increasing flow rate; when the flow rate is 1.5–2.0 m / s, the extraction rate tends to stabilize (maximum value); when the flow rate is >2.5 m / s, energy consumption increases dramatically and the medicinal powder is stirred up, making filtration difficult. Therefore, 1.5–2.0 m / s is the optimal economic flow rate.

[0086] Example 9: Optimization of Ceramic Membrane Cleaning Process Different cleaning agents (NaOH, HNO3, NaClO) and cleaning frequencies were used.

[0087] Results: The flux recovery rate of simple water washing was only 60%; by using 0.1% NaOH + 0.05% NaClO, with a pulse backwash of 30 seconds every 15 minutes, the flux recovery rate was >95%, and the membrane life was extended to more than 2 years.

[0088] Example 10: Pilot-scale verification Three batches of pilot-scale production were carried out in a 1000L extraction tank, with the same process parameters as in Example 1.

[0089] Batch data: Batch 1: Saikosaponin a+d 13.1mg / 100ml, Paeoniflorin 28.2mg / 100ml.

[0090] Batch 2: Saikosaponin a+d 13.4mg / 100ml, Paeoniflorin 28.8mg / 100ml.

[0091] Batch 3: Saikosaponin a+d 13.0mg / 100ml, Paeoniflorin 28.0mg / 100ml.

[0092] Conclusion: The RSD is all <2%, indicating that the process of this invention has excellent reproducibility and is suitable for large-scale industrial production.

[0093] Conclusion: The Chai Shao oral liquid prepared by this invention is significantly superior to the prior art in terms of chemical component content, physical stability and pharmacological activity.

[0094] The above are merely preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A method for preparing a Chai Shao oral liquid, characterized in that, Includes the following steps: Step 1: Pre-processing and cleaning of medicinal materials. Weigh out the following ingredients according to the prescription: Bupleurum, white peony root, immature bitter orange, and licorice. Bupleurum needs to be processed with vinegar, and white peony root needs to be stir-fried with wine. Clean each medicinal material, removing impurities and non-medicinal parts, and drain after washing. Step 2: Differentiated ultrafine grinding. The cleaned medicinal materials are placed in an airflow ultrafine grinder for grinding. The feeding speed is controlled at 10-15 kg / h and the grinding pressure is 0.6-0.8 MPa. Bupleurum and Citrus aurantium are ground to a cell wall breakage rate ≥95% and a particle size distribution D90 ≤15 μm; Paeonia lactiflora and Glycyrrhiza uralensis are ground to a cell wall breakage rate ≥90% and a particle size distribution D90 ≤25 μm. Step 3: Gradient temperature soaking and softening. Mix the pulverized medicinal materials evenly according to the prescription ratio, add 6-8 times the total weight of the medicinal materials in purified water, place it in an extraction tank with a jacket, and use a gradient temperature method for temperature soaking: first, stir and soak at 30-35℃ for 20-30 minutes, then raise the temperature to 50-55℃ at a rate of 1-2℃ / min, and continue to soak for 10-15 minutes until the medicinal materials have fully absorbed water and swelled. Step 4: Variable temperature countercurrent dynamic extraction. Add 8-10 times the amount of purified water to the soaked medicinal materials. Turn on the variable frequency agitator and control the speed at 60-80 r / min. Use a variable temperature extraction process: the first stage is extraction at 75-80℃ for 40-50 minutes, and the second stage is heating to 90-95℃ and holding for 30-40 minutes. During the extraction process, the extract is drawn from the bottom of the tank by a circulating pump, heated by a plate heat exchanger, and then sprayed tangentially from the top of the tank to form dynamic turbulence. The circulation velocity is 1.5-2.0 m / s. Step 5: Ultrasonic field-assisted dissolution. In the second stage of temperature-controlled extraction, the embedded ultrasonic generator is simultaneously activated, with the ultrasonic frequency set to 25-35kHz and the sound intensity density controlled at 0.5-1.0W / cm². 2 The action time is synchronized with the extraction time in the second stage; Step 6: Ceramic membrane microfiltration for impurity removal. After extraction, the extract is pumped into a ceramic membrane filtration system. First, it passes through a microfiltration membrane with a pore size of 0.2 μm to remove fine particles and bacteria, and then through an ultrafiltration membrane with a molecular weight cutoff of 5000 Da to remove large molecular impurities such as polysaccharides and proteins. The permeate is then collected. Step 7: Low-temperature vacuum concentration. The permeate is pumped into a falling film evaporator and concentrated under vacuum conditions of -0.085 to -0.095 MPa and feed temperature of 45-55℃ until the relative density reaches 1.10-1.15 (measured at 60℃). Step 8: Alcohol precipitation and preparation. Add 95% ethanol to the concentrate to make the alcohol content reach 60%-65%. Stir well and let stand in the refrigerator for 12-24 hours. Take the supernatant, recover the ethanol and adjust the pH value to 4.5-5.

5. Add flavoring agent. Step 9: Filling and sterilization. After the prepared medicine solution is filtered through a 0.22μm filter, it is filled into bottles and sterilized by moist heat at 115℃ for 30 minutes to obtain Chai Shao Oral Liquid.

2. The method for preparing a Chai Shao oral liquid according to claim 1, characterized in that, The difference in particle size between Bupleurum chinense, Citrus aurantium, Paeonia lactiflora, and Glycyrrhiza uralensis in step two is based on the different textures of the medicinal materials: Bupleurum chinense and Citrus aurantium are hard and fibrous, requiring a higher cell wall breaking rate to release saikosaponins and synephrine; Paeonia lactiflora and Glycyrrhiza uralensis are relatively loose in texture, and moderate pulverization is necessary to avoid excessive adsorption of active ingredients.

3. The method for preparing a Chai Shao oral liquid according to claim 1, characterized in that, The specific control logic of the "variable temperature countercurrent dynamic extraction" described in step four is as follows: the first stage of low temperature long-time extraction aims to dissolve saponin components with poor heat sensitivity, and the second stage of high temperature short-time extraction aims to dissolve flavonoids and aglycone components with better heat resistance. The dynamic circulation avoids the channeling phenomenon caused by the compaction of the medicinal material layer.

4. The method for preparing a Chai Shao oral liquid according to claim 1, characterized in that, The ultrasonic generator mentioned in step five is a frequency conversion type. Its frequency is sinusoidally modulated during the extraction process, with a modulation period of 10-20 seconds and a frequency scanning between 20-40kHz to avoid extraction dead zones caused by standing wave effects.

5. The method for preparing a Chai Shao oral liquid according to claim 1, characterized in that, The ceramic membrane mentioned in step six is ​​an alumina / zirconia composite membrane with a membrane pore size tolerance controlled within ±0.01μm and a filtration transmembrane pressure difference (TMP) controlled within 0.1-0.3MPa. It is cleaned by pulse backwashing with a backwashing frequency of once every 15 minutes and a backwashing time of 30 seconds.

6. The method for preparing a Chai Shao oral liquid according to claim 1, characterized in that, The low-temperature vacuum concentration described in step (7) uses triple-effect falling film evaporation, with the first effect temperature at 65-70℃, the second effect temperature at 55-60℃, and the third effect temperature at 45-50℃. The retention rate of the effective components is increased by more than 15% compared with single-effect concentration.

7. The method for preparing a Chai Shao oral liquid according to claim 1, characterized in that, The flavoring agent is a complex of xylitol and steviol glycosides in a weight ratio of 10:1, and is added at 2%-4% of the total amount of the medicinal liquid to mask the bitterness of Bupleurum and the sweetness of licorice and improve the taste.

8. The method for preparing a Chai Shao oral liquid according to claim 1, characterized in that, The process for vinegar-processed Bupleurum in step one is as follows: Take Bupleurum slices, mix them with rice vinegar, let them soak until the vinegar is absorbed, place them in a stir-frying container, heat them over a low heat, stir-fry until dry, remove and let cool; use 15kg of rice vinegar for every 100kg of Bupleurum slices. The process for wine-processed Paeonia lactiflora is as follows: Take Paeonia lactiflora slices, mix them with rice wine, let them soak until the wine is absorbed, place them in a stir-frying container, heat them over a low heat, stir-fry until slightly yellow, remove and let cool; use 10kg of rice wine for every 100kg of Paeonia lactiflora slices.