In-situ inclusion preparation method of a traditional Chinese medicine compound danpi phenol inclusion compound

Abstract

The application provides a preparation method of a complex in a traditional Chinese medicine compound, and belongs to the technical field of medicine industry. The application directly carries out inclusion on a distillate containing complex components such as paeonol, uses hydroxypropyl betadex as an inclusion agent, optimizes the ratio of the distillate containing complex components such as paeonol and the inclusion agent, and directly prepares paeonol inclusion compounds with good stability and not easy to separate from traditional Chinese medicine compound decoctions by directly freeze-drying. The inclusion rate can reach more than 80%, and the yield of the inclusion compound can reach more than 95%. The preparation method not only simplifies the extraction process of the traditional Chinese medicine compound containing paeonia suffruticosa and reduces the cost, but also has a wide application prospect.

Description

Technical Field

[0001] This invention relates to the pharmaceutical technology industry, specifically to an in-situ inclusion complex preparation method for paeonol inclusion compounds in traditional Chinese medicine compound preparations. Background Technology

[0002] Paeonol is one of the main active ingredients in the root bark of *Paeonia suffruticosa* Andr. (Ranunculaceae family) and the dried root and rhizome of *Cynanchumpaniculatum* (Bge.) Kitag. (Asclepiadaceae family). Paeonol is a white or slightly yellow crystalline powder with a characteristic odor and a slightly spicy taste. It is readily soluble in ethanol and methanol, soluble in hot water, insoluble in cold water, and volatilizes with steam. Paeonol possesses various pharmacological activities, such as antipyretic analgesia, anti-inflammatory and antibacterial effects, anti-allergic properties, cardiovascular protection, antitumor activity, and hypoglycemic effects. Paeonol exhibits significant pharmacological activity, few adverse reactions, and a short half-life, making it a promising candidate for a wide range of applications. Because paeonol has a distinctive odor, is highly volatile, unstable to light, temperature, and humidity, and has poor water solubility, it is often treated by inclusion complexation to reduce its volatility, increase its water solubility, improve its stability and bioavailability. Currently, common methods for preparing paeonol inclusion complexes include saturated aqueous solution method, ultrasonic method, grinding method, and spray drying method.

[0003] Existing processes for paeonol inclusion complexes typically involve decocting peony bark alone, extracting paeonol distillate using steam distillation, refrigerating the collected distillate overnight to obtain paeonol crystals, dissolving the crystals in ethanol, and then inclusion complexes with a saturated aqueous solution of beta-cyclodextrin. However, when using steam distillation to co-decoct complexes containing peony bark, the resulting distillate contains complex components such as paeonol. Due to the low paeonol content in the distillate, refrigeration fails to yield paeonol crystals. Directly including the mixed paeonol distillate with beta-cyclodextrin in the prepared paeonol inclusion complex suffers from low inclusion rate and yield, instability, and poor water solubility. To address the issue of low yield and inclusion rate of paeonol inclusion complex during the co-decoction of traditional Chinese medicine formulas containing peony bark, this invention provides a one-step method for directly preparing paeonol inclusion complexes from the steam distillate of co-decoctions of traditional Chinese medicine formulas containing peony bark. The prepared paeonol inclusion complexes exhibit high inclusion rate and yield, as well as good stability. Summary of the Invention

[0004] The purpose of this invention is to provide an in-situ inclusion preparation method for paeonol inclusion complex in traditional Chinese medicine compound preparations. The prepared paeonol inclusion complex has good stability, is not easily detached, and has a high inclusion rate and yield.

[0005] The technical solution of this invention is implemented as follows: This invention provides an in-situ inclusion complex preparation method for paeonol inclusion compounds in traditional Chinese medicine compound preparations, comprising the following steps: (1) Take all the medicinal materials in the compound containing peony bark, add 6 to 15 times the amount of water and soak for 0.5 to 2 hours, then extract by steam distillation and collect 1 to 5 times the weight of the medicinal materials in the distillate; (2) Keep the distillate at 25-50℃ and stir. Add hydroxypropyl betacyclodextrin to the distillate. The mass ratio of the distillate to hydroxypropyl betacyclodextrin is 5:1 to 50:1. (3) Continue stirring to dissolve the hydroxypropyl betacyclodextrin, and keep it at 25-50°C for 0.5-4 hours. After stopping the heating, continue stirring until the temperature reaches room temperature to obtain the inclusion solution. (4) Freeze-dry the inclusion solution to obtain paeonol inclusion complex.

[0006] Preferably, the conditions for steam distillation extraction in step (1) are: add 10 times the amount of water to soak for 1 hour, and collect 3 times the weight of the medicinal material in the distillate.

[0007] Preferably, hydroxypropyl betacyclodextrin is used as an inclusion agent in step (2).

[0008] Preferably, the mass ratio of the distillate to hydroxypropyl betacyclodextrin in step (2) is 30:1.

[0009] Preferably, the stirring conditions in step (3) are stirring at 30°C for 1 hour.

[0010] The present invention also provides a paeonol inclusion complex prepared by the above method.

[0011] Beneficial effects: 1. This invention utilizes hydroxypropyl betacyclodextrin as an inclusion agent, limiting the ratio of paeonol distillate to inclusion agent to 5:1 to 50:1 (g / g), and employs direct freeze-drying to achieve in-situ inclusion of paeonol in the steam distillate of traditional Chinese medicine compound decoctions containing peony bark. This inclusion process solves key issues such as inclusion rate, yield, and stability of the active ingredient, while eliminating several energy-intensive and easily destructive steps required in the production of traditional Chinese medicine compound formulas containing peony bark, including separate extraction of peony bark distillate, purification of the distillate, and subsequent paeonol inclusion. This invention reduces the exposure and loss of paeonol in intermediate stages, thereby preparing a stable and non-detachable paeonol inclusion complex, thus improving the inclusion rate and yield. The inclusion rate can reach over 80%, and the inclusion complex yield over 95%, ensuring the full utilization of the effective components in the traditional Chinese medicine compound. Meanwhile, the inclusion of paeonol can be completed directly in one step by steam distillation of the decoction of traditional Chinese medicine compound. The reduction of preparation steps directly leads to a decrease in equipment, time and energy costs, and improves overall efficiency and utilization.

[0012] 2. The method of the present invention does not depend on a specific compound and has good applicability to various compound formulas containing peony bark, and the process has strong universality.

[0013] 3. The steam distillation and freeze drying technologies used in this invention are mature pharmaceutical preparation technologies. Their preparation process conditions are mild, the operation is simple, the repeatability is good, and no complex and expensive equipment is required. They are suitable for large-scale industrial production and have a high market transformation prospect. Detailed Implementation

[0014] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0015] To enable those skilled in the art to fully understand the present invention, the present invention is further illustrated below through specific embodiments. However, those skilled in the art should understand that the embodiments of the present invention do not limit the present invention in any way.

[0016] Example 1 (1) According to the prescription ratio of the classic formula "Liuwei Dihuang Wan" included in the Chinese Pharmacopoeia, take 96g of prepared Rehmannia root, 48g of Cornus officinalis, 36g of Paeonia suffruticosa, 48g of Dioscorea opposita, 36g of Poria cocos, and 36g of Alisma plantago-aquatica, totaling 300g. Add 10 times the amount of water, soak for 1 hour, heat to a slight boil, extract by steam distillation, and collect 900mL of Paeonia suffruticosa distillate.

[0017] (2) Preparation of paeonol inclusion complex: Take 750g of the paeonol distillate described in (1), place it in a 30℃ magnetic stirring water bath and keep it warm and stirred. Add a certain amount of hydroxypropyl beta-cyclodextrin directly to the paeonol distillate [the ratio of paeonol distillate to hydroxypropyl beta-cyclodextrin is 30:1 (g / g)], stir to dissolve the hydroxypropyl beta-cyclodextrin, stir at 30℃ for 1h, stop heating, continue stirring until room temperature, and obtain the inclusion solution. Freeze-dry the inclusion solution to obtain 24.21g of inclusion complex.

[0018] (3) Content determination Chromatographic conditions and system suitability test: Octadecylsilane-bonded silica gel was used as the stationary phase; methanol-water (48:52) was used as the mobile phase; the detection wavelength was 274 nm; the theoretical plate number calculated based on the paeonol peak should not be less than 6000.

[0019] Preparation of reference solution: Accurately weigh an appropriate amount of paeonol reference standard, add methanol to prepare a solution containing 60 μg per milliliter.

[0020] Preparation of the test solution: Take 2g of paeonol distillate or about 0.2g of paeonol inclusion complex, accurately weigh it, place it in a stoppered conical flask, accurately add 25ml of methanol, stopper tightly, shake well, weigh, sonicate (power 500W, frequency 50kHz) for 30 minutes, cool, weigh again, make up the lost weight with methanol, shake well, filter, and the test solution is obtained.

[0021] Determination method: Accurately pipette 10 μl of the reference solution and the test solution into the liquid chromatograph, measure and calculate to obtain the result.

[0022] Weigh out appropriate amounts of paeonol distillate and paeonol inclusion complex, and determine the content of the distillate and inclusion complex according to the above-mentioned content determination method, and calculate the inclusion rate of paeonol.

[0023] Paeonol inclusion rate (%) = (weight of inclusion compound × paeonol content in inclusion compound) ÷ amount of paeonol in distillate × 100% Inclusion complex yield (%) = Inclusion complex weight ÷ (Amount of hydroxypropyl betacyclodextrin added + Amount of paeonol in distillate) × 100% Calculations showed that the paeonol content in the inclusion complex was 1.856%, the inclusion rate of paeonol was 92.17%, and the yield of the inclusion complex was 96.84%.

[0024] Example 2 (1) According to the prescription ratio of the classic formula "Guizhi Fuling Wan" included in the Chinese Pharmacopoeia, take 30g of cinnamon twig, 30g of poria cocos, 30g of peony bark, 30g of red peony root, and 30g of peach kernel, totaling 150g. Add 10 times the amount of water, soak for 1 hour, heat to a slight boil, extract by steam distillation, and collect 450mL of paeonol distillate.

[0025] (2) Preparation of paeonol inclusion complex: Take 375g of the paeonol distillate described in (1), place it in a 30℃ magnetic stirring water bath and keep it warm and stirred. Add a certain amount of hydroxypropyl beta-cyclodextrin directly to the paeonol distillate [the ratio of paeonol distillate to hydroxypropyl beta-cyclodextrin is 30:1 (g / g)], stir to dissolve the hydroxypropyl beta-cyclodextrin, stir at 30℃ for 1h, stop heating, continue stirring until room temperature, and obtain the inclusion solution. Freeze-dry the inclusion solution to obtain 11.95g of inclusion complex.

[0026] Weigh appropriate amounts of paeonol distillate and paeonol inclusion complex, and determine the content of paeonol distillate and paeonol inclusion complex according to the method under "Content Determination" in Example 1, and calculate the inclusion rate of paeonol. The calculation methods for paeonol inclusion rate and inclusion complex yield are the same as in Example 1.

[0027] Calculations showed that the paeonol content in the inclusion complex was 1.868%, the inclusion rate of paeonol was 91.58%, and the yield of the inclusion complex was 95.60%.

[0028] Example 3: (1) According to the formula of a new Chinese medicine compound containing Scutellaria baicalensis, Paeonia suffruticosa, Paeonia lactiflora, Gardenia jasminoides, etc., take 50g of Scutellaria baicalensis, 45g of Paeonia suffruticosa, 35g of Paeonia lactiflora, 30g of Gardenia jasminoides, 30g of Citrus reticulata, 30g of Pinellia ternata, and 30g of Glycyrrhiza uralensis, totaling 250g. Add 10 times the amount of water, soak for 1 hour, heat to a slight boil, extract by steam distillation, and collect 750mL of Paeonia lactiflora distillate.

[0029] (2) Preparation of paeonol inclusion complex: Take 600g of the paeonol distillate described in (1), place it in a 30℃ magnetic stirring water bath and keep it warm and stirred. Add a certain amount of hydroxypropyl beta-cyclodextrin directly to the paeonol distillate [the ratio of paeonol distillate to hydroxypropyl beta-cyclodextrin is 30:1 (g / g)], stir to dissolve the hydroxypropyl beta-cyclodextrin, stir at 30℃ for 1h, stop heating, continue stirring until room temperature, and obtain the inclusion solution. Freeze-dry the inclusion solution to obtain 19.30g of inclusion complex.

[0030] Weigh appropriate amounts of paeonol distillate and paeonol inclusion complex, and determine the content of paeonol distillate and paeonol inclusion complex according to the method under "Content Determination" in Example 1, and calculate the inclusion rate of paeonol. The calculation methods for paeonol inclusion rate and inclusion complex yield are the same as in Example 1.

[0031] Calculations showed that the paeonol content in the inclusion complex was 1.904%, the inclusion rate of paeonol was 92.91%, and the yield of the inclusion complex was 95.15%.

[0032] Example 4 (1) Extraction of paeonol distillate: According to the formulation of the compound prescription of the new Chinese medicine containing peony bark in Example 3, take a total of 250g of compound medicine containing peony bark, add 6 times the amount of water, soak for 1 hour, heat to a slight boil, extract by steam distillation, and collect 250mL of paeonol distillate.

[0033] (2) Preparation of paeonol inclusion complex: Take 200g of the paeonol distillate described in (1), place it in a 30℃ magnetic stirring water bath and keep it warm and stirred. Add a certain amount of hydroxypropyl beta-cyclodextrin directly to the paeonol distillate [the ratio of paeonol distillate to hydroxypropyl beta-cyclodextrin is 5:1 (g / g)], stir to dissolve the hydroxypropyl beta-cyclodextrin, stir at 30℃ for 1h, stop heating, continue stirring until room temperature, and obtain the inclusion solution. Freeze-dry the inclusion solution to obtain 38.52g of inclusion complex.

[0034] Weigh appropriate amounts of paeonol distillate and paeonol inclusion complex, and determine the content of paeonol distillate and paeonol inclusion complex according to the method under "Content Determination" in Example 1, and calculate the inclusion rate of paeonol. The calculation methods for paeonol inclusion rate and inclusion complex yield are the same as in Example 1.

[0035] Calculations showed that the paeonol content in the inclusion complex was 0.316%, the inclusion rate of paeonol was 93.63%, and the yield of the inclusion complex was 96.30%.

[0036] Example 5 (1) According to the formulation of the compound prescription of the new Chinese medicine containing peony bark in Example 3, take a total of 250g of compound medicine containing peony bark, add 15 times the amount of water, soak for 1 hour, heat to a slight boil, extract by steam distillation, and collect 1250mL of peony bark extract.

[0037] (2) Preparation of paeonol inclusion complex: Take 1000g of the paeonol distillate described in (1), place it in a 30℃ magnetic stirring water bath and keep it warm and stirred. Add a certain amount of hydroxypropyl beta-cyclodextrin directly to the paeonol distillate [the ratio of paeonol distillate to hydroxypropyl beta-cyclodextrin is 50:1 (g / g)], stir to dissolve the hydroxypropyl beta-cyclodextrin, stir at 30℃ for 1h, stop heating, continue stirring until room temperature, and obtain the inclusion solution. Freeze-dry the inclusion solution to obtain 19.14g of inclusion complex.

[0038] Weigh appropriate amounts of paeonol distillate and paeonol inclusion complex, and determine the content of paeonol distillate and paeonol inclusion complex according to the method under "Content Determination" in Example 1, and calculate the inclusion rate of paeonol. The calculation methods for paeonol inclusion rate and inclusion complex yield are the same as in Example 1.

[0039] Calculations showed that the paeonol content in the inclusion complex was 2.845%, the inclusion rate of paeonol was 83.77%, and the yield of the inclusion complex was 95.70%.

[0040] Example 6: Investigation of different soaking times (1) Decoction According to the formulation of the new traditional Chinese medicine compound prescription containing peony bark in Example 3, ... ① Take a total of 250g of compound medicinal materials containing peony bark, add 10 times the amount of water, heat to a gentle boil, extract by steam distillation, and collect 750mL of peony bark extract.

[0041] ② Take a total of 250g of compound medicinal materials containing peony bark, add 10 times the amount of water, soak for 0.5h, heat to a gentle boil, extract by steam distillation, and collect 750mL of peony bark extract.

[0042] ③ Take a total of 250g of compound medicinal materials containing peony bark, add 10 times the amount of water, soak for 1 hour, heat to a gentle boil, extract by steam distillation, and collect 750mL of peony bark extract.

[0043] ④ Take a total of 250g of compound medicinal materials containing peony bark, add 10 times the amount of water, soak for 2 hours, heat to a gentle boil, extract by steam distillation, and collect 750mL of peony bark phenol distillate.

[0044] (2) Preparation of paeonol inclusion complex Take 600g of each of the paeonol distillate from items ①, ②, ③, and ④ in (1), place them in a 30℃ magnetic stirring water bath and keep them warm and stirred. Add a certain amount of hydroxypropyl beta-cyclodextrin directly to the paeonol distillate [the ratio of paeonol distillate to beta-cyclodextrin is 30:1 (g / g)], stir to dissolve the hydroxypropyl beta-cyclodextrin, stir at a constant temperature of 30℃ for 1 hour, stop heating, continue stirring until room temperature, freeze dry the inclusion solutions to obtain inclusion complex ① 19.24g, inclusion complex ② 19.18g, inclusion complex ③ 19.30g, and inclusion complex ④ 19.28g respectively.

[0045] Weigh appropriate amounts of paeonol distillate and paeonol inclusion complexes ①, ②, ③, and ④. Determine the content of the inclusion complexes according to the method described in "Content Determination" of Example 1, and calculate the inclusion rate of paeonol. The calculation methods for the paeonol inclusion rate and the yield of the inclusion complexes are the same as in Example 1.

[0046] The effects of different soaking times on the inclusion process were calculated, with the paeonol content in the inclusion compound, the yield of the inclusion compound, and the inclusion rate as evaluation indicators. The results showed that the paeonol content in the inclusion compound increased significantly after soaking. There was no significant difference in the effect between soaking for 1 hour and 2 hours, while the inclusion effect after soaking for 1 hour was close to the optimal value.

[0047] The experimental results are shown in the table below.

[0048]

[0049] Example 7: Investigation of different proportions of inclusion agents (1) According to the formulation of the compound prescription of the new Chinese medicine containing peony bark in Example 3, take a total of 750g of compound medicine containing peony bark, add 10 times the amount of water, soak for 1 hour, heat to a slight boil, extract by steam distillation, and collect 2250mL of peony bark extract.

[0050] (2) Preparation of paeonol inclusion complex: Take 300g of each of the paeonol distillate from (1), a total of 6 portions, and place them in a 30℃ magnetic stirring water bath for heat preservation and stirring. Add a certain amount of hydroxypropyl beta-cyclodextrin directly to the paeonol distillate: ① The ratio of paeonol distillate to hydroxypropyl beta-cyclodextrin is 5:1 (g / g); ② The ratio of paeonol distillate to hydroxypropyl beta-cyclodextrin is 10:1 (g / g); ③ The ratio of paeonol distillate to hydroxypropyl beta-cyclodextrin is 20:1 (g / g); ④ The ratio of paeonol distillate to hydroxypropyl beta-cyclodextrin is 30:1. :1 (g / g); ⑤ The ratio of paeonol distillate to hydroxypropyl beta-cyclodextrin is 40:1 (g / g); ⑥ The ratio of paeonol distillate to hydroxypropyl beta-cyclodextrin is 50:1 (g / g). Stir to dissolve hydroxypropyl beta-cyclodextrin, keep warm at 30℃ for 1 hour, stop heating, continue stirring to room temperature, and obtain inclusion solution. Freeze-dry the inclusion solution to obtain: inclusion compound ① 57.72g; inclusion compound ② 28.62g; inclusion compound ③ 14.34g; inclusion compound ④ 9.64g; inclusion compound ⑤ 7.20g; inclusion compound ⑥ 5.70g.

[0051] Weigh appropriate amounts of paeonol distillate and paeonol inclusion complex, and determine the content of paeonol distillate and paeonol inclusion complex according to the method under "Content Determination" in Example 1, and calculate the inclusion rate of paeonol. The calculation methods for paeonol inclusion rate and inclusion complex yield are the same as in Example 1.

[0052] Calculations showed that the yields of the inclusion compounds obtained from the six groups with different inclusion agent ratios were all higher than 95%, and the inclusion rates were all higher than 80%.

[0053] The experimental results are shown in the table below.

[0054]

[0055] Example 8 Inclusion Temperature Investigation (1) According to the formulation of the compound prescription of the new Chinese medicine containing peony bark in Example 3, take a total of 250g of compound medicine containing peony bark, add 10 times the amount of water, soak for 1 hour, heat to a slight boil, extract by steam distillation, and collect 750mL of peony bark phenol distillate.

[0056] (2) Preparation of paeonol inclusion complex: Take 150g of each of the paeonol distillate from (1), a total of 4 portions, and place them in a magnetic stirring water bath at ①25℃, ②30℃, ③40℃, and ④50℃ respectively for heating and stirring. Add a certain amount of hydroxypropyl beta-cyclodextrin directly to the paeonol distillate [the ratio of paeonol distillate to hydroxypropyl beta-cyclodextrin is 30:1 (g / g)]. Stir to dissolve the hydroxypropyl beta-cyclodextrin. Stir at ①25℃, ②30℃, ③40℃, and ④50℃ for 1 hour respectively. Stop heating and continue stirring until room temperature to obtain inclusion solution. Freeze-dry the inclusion solution to obtain inclusion complex ① 4.78g; inclusion complex ② 4.83g; inclusion complex ③ 4.75g; inclusion complex ④ 4.76g.

[0057] Weigh appropriate amounts of paeonol distillate and paeonol inclusion complex, and determine the content of paeonol distillate and paeonol inclusion complex according to the method under "Content Determination" in Example 1, and calculate the inclusion rate of paeonol. The calculation methods for paeonol inclusion rate and inclusion complex yield are the same as in Example 1.

[0058] Calculations showed that there was no significant difference in inclusion rate when the inclusion temperature was between 25-40℃, while the inclusion rate decreased slightly at 50℃.

[0059] The experimental results are shown in the table below.

[0060]

[0061] Example 9: Inclusion Time Examination (1) According to the formulation of the compound prescription of the new Chinese medicine containing peony bark in Example 3, take 250g of compound medicine containing peony bark, add 10 times the amount of water, soak for 1 hour, heat to a slight boil, extract by steam distillation, and collect 750mL of peony bark phenol distillate.

[0062] (2) Preparation of paeonol inclusion complex: Take 150g of each of the paeonol distillate from (1), for a total of 4 portions; place them in a 30℃ magnetic stirring water bath and keep them warm and stirred. Add a certain amount of hydroxypropyl beta-cyclodextrin directly to the paeonol distillate [the ratio of paeonol distillate to hydroxypropyl beta-cyclodextrin is 30:1 (g / g)] and stir to dissolve the hydroxypropyl beta-cyclodextrin. Stir at 30℃ for ①0.5h, ②1h, ③2h, and ④4h respectively. Stop heating and continue stirring until room temperature to obtain inclusion solution. Freeze-dry the inclusion solution to obtain inclusion complex ①4.75g; inclusion complex ②4.80g; inclusion complex ③4.79g; and inclusion complex ④4.82g.

[0063] Weigh appropriate amounts of paeonol distillate and paeonol inclusion complex, and determine the content of paeonol distillate and paeonol inclusion complex according to the method under "Content Determination" in Example 1, and calculate the inclusion rate of paeonol. The calculation methods for paeonol inclusion rate and inclusion complex yield are the same as in Example 1.

[0064] Calculations showed that the inclusion time of 0.5h to 4h had no significant impact on the inclusion rate.

[0065] The experimental results are shown in the table below.

[0066]

[0067] Comparative Example 1 (1) According to the formulation of the compound traditional Chinese medicine containing peony bark in Example 3, take a total of 750g of compound medicinal materials containing peony bark, add 10 times the amount of water, soak for 1h, heat to a slight boil, extract by steam distillation, and collect 2250mL of paeonol distillate. Take 1800g of paeonol distillate, place it on a 30℃ magnetic stirring water bath and keep it warm and stirred, add a certain amount of hydroxypropyl betacyclodextrin directly to the paeonol distillate [the ratio of paeonol distillate to hydroxypropyl betacyclodextrin is 30:1 (g / g)], stir to dissolve the hydroxypropyl betacyclodextrin, keep it warm at 30℃ and stir for 1h, stop heating, continue stirring to room temperature, and obtain inclusion solution.

[0068] (2) Divide the inclusion solution obtained in (1) into three equal parts and dry them respectively using the following drying methods: ① The inclusion solution was directly freeze-dried to obtain 19.25g of inclusion compound.

[0069] ② The inclusion solution was concentrated to 300 ml, and the concentrate was freeze-dried to obtain 18.82 g of inclusion compound.

[0070] ③ Concentrate the inclusion solution to 300 ml, and dry the concentrate under reduced pressure (≤75℃) to obtain 18.28 g of inclusion compound.

[0071] Weigh appropriate amounts of paeonol distillate and paeonol inclusion complexes ①, ②, and ③. Determine the content of paeonol in the distillate and inclusion complexes according to the method described in "Content Determination" of Example 1, and calculate the inclusion rate of paeonol. The calculation methods for the paeonol inclusion rate and inclusion complex yield are the same as in Example 1.

[0072] Calculations show that direct freeze-drying of the inclusion solution yields the best results.

[0073] The experimental results are shown in the table below.

[0074]

[0075] Comparative Example 2 (1) According to the formulation of the compound prescription of the new Chinese medicine containing peony bark in Example 3, take a total of 750g of compound medicine containing peony bark, add 10 times the amount of water, soak for 1 hour, heat to a slight boil, extract by steam distillation, and collect 2250mL of peony bark extract.

[0076] (2) Preparation of paeonol inclusion complex: ① Take 600g of the paeonol distillate described in (1), place it in a 30℃ magnetic stirring water bath and keep it warm and stirred. Add a certain amount of beta-cyclodextrin directly to the paeonol distillate [the ratio of paeonol distillate to beta-cyclodextrin is 30:1 (g / g)], stir to dissolve the beta-cyclodextrin, stir at a constant temperature of 30℃ for 1h, stop heating, continue stirring until room temperature, let the inclusion solution settle at low temperature for 36h, filter, and dry the solid at 40℃ with hot air to obtain 11.23g of inclusion complex.

[0077] ② Take 600g of the paeonol distillate described in (1), place it in a 30℃ magnetic stirring water bath and keep it warm and stirred. Add a certain amount of beta-cyclodextrin directly to the paeonol distillate [the ratio of paeonol distillate to beta-cyclodextrin is 30:1 (g / g)], stir to dissolve the beta-cyclodextrin, stir at a constant temperature of 30℃ for 1h, stop heating, continue stirring until room temperature, freeze dry the inclusion solution to obtain 19.14g of inclusion complex.

[0078] ③ Take 600g of the paeonol distillate described in (1), place it in a 30℃ magnetic stirring water bath and keep it warm and stirred. Add a certain amount of hydroxypropyl beta-cyclodextrin directly to the paeonol distillate [the ratio of paeonol distillate to hydroxypropyl beta-cyclodextrin is 30:1 (g / g)], stir to dissolve the hydroxypropyl beta-cyclodextrin, stir at 30℃ for 1h, stop heating, continue stirring until room temperature, freeze dry the inclusion solution to obtain 19.28g of inclusion complex.

[0079] Weigh appropriate amounts of paeonol distillate and paeonol inclusion complexes ①, ②, and ③. Determine the content of the inclusion complexes according to the method described in "Content Determination" of Example 1, and calculate the inclusion rate of paeonol. The calculation methods for the paeonol inclusion rate and the yield of the inclusion complexes are the same as in Example 1.

[0080] Calculations and experimental results show that hydroxypropyl betacyclodextrin is the best inclusion agent, and direct freeze-drying of the inclusion solution yields the best results.

[0081] The experimental results are shown in the table below.

[0082]

[0083] Comparative Example 3 (1) Decoction ①According to the formulation of the compound traditional Chinese medicine containing peony bark in Example 3, take 500g of the compound traditional Chinese medicine containing peony bark, add 10 times the amount of water, soak for 1 hour, heat to a gentle boil, extract by steam distillation, and collect 1500mL of peony bark extract for later use.

[0084] ② Take 180g of peony bark, which is twice the amount in item ①, add 10 times the amount of water, soak for 1 hour, heat to a gentle boil, extract by steam distillation, and collect 540mL of peony bark phenol distillate for later use.

[0085] (2) Preparation of paeonol inclusion complex Take 1200g and 216g of the paeonol distillate from items ① and ② in (1) respectively, place them in a 30℃ magnetic stirring water bath and keep them warm and stirred. Add a certain amount of hydroxypropyl beta-cyclodextrin directly to the paeonol distillate [the ratio of paeonol distillate to beta-cyclodextrin is 30:1 (g / g)], stir to dissolve the hydroxypropyl beta-cyclodextrin, stir at a constant temperature of 30℃ for 1h, stop heating, continue stirring until room temperature, freeze dry the inclusion solutions respectively to obtain inclusion compound ① 38.56g and inclusion compound ② 6.90g respectively.

[0086] Take 216g of the paeonol distillate from item ② in (1), let it stand and refrigerate at 4℃ for 48h, filter it to obtain paeonol crystals, dissolve the paeonol crystals in 5ml of anhydrous ethanol to obtain paeonol ethanol solution, add the paeonol ethanol solution dropwise to the aqueous solution of hydroxypropyl beta-cyclodextrin [150g of water, 7.2g of hydroxypropyl beta-cyclodextrin, stir to dissolve the hydroxypropyl beta-cyclodextrin], stir at a constant temperature of 30℃ for 1h, stop heating, continue stirring to room temperature, freeze dry the inclusion solution to obtain inclusion compound ③ 6.88g.

[0087] Weigh appropriate amounts of paeonol distillate and paeonol inclusion complexes ①, ②, and ③. Determine the content of the inclusion complexes according to the method described in "Content Determination" of Example 1, and calculate the inclusion rate of paeonol. The calculation methods for the paeonol inclusion rate and the yield of the inclusion complexes are the same as in Example 1.

[0088] Calculations showed that the contents of paeonol in inclusion complexes ①, ②, and ③ were 1.832%, 15.284%, and 11.633%, respectively, with inclusion rates of 90.57%, 93.89%, and 71.26%, and inclusion complex yields of 96.4%, 95.8%, and 95.6%, respectively.

[0089] The experimental results are shown in the table below.

[0090]

[0091] Experimental Example: Effect of Paeonol Inclusion Complex Stability on Experiment (1) According to the formulation of the compound prescription of traditional Chinese medicine containing peony bark in Example 3, take a total of 1100g of traditional Chinese medicine compound containing peony bark, add 10 times the amount of water, soak for 1 hour, heat to a slight boil, extract by steam distillation, and collect 3300mL of peony bark extract.

[0092] (2) Preparation of paeonol inclusion complex: Take 3000g of the paeonol distillate described in (1), place it in a 30℃ magnetic stirring water bath and keep it warm and stirred. Add a certain amount of hydroxypropyl beta-cyclodextrin directly to the paeonol distillate [the ratio of paeonol distillate to hydroxypropyl beta-cyclodextrin is 30:1 (g / g)], stir to dissolve the hydroxypropyl beta-cyclodextrin, stir at 30℃ for 1h, stop heating, continue stirring until room temperature, and obtain the inclusion solution. Freeze-dry the inclusion solution to obtain 97.1g of paeonol inclusion complex.

[0093] Weigh appropriate amounts of paeonol distillate and paeonol inclusion complex, and determine the content of paeonol distillate and paeonol inclusion complex according to the method under "Content Determination" in Example 1, and calculate the inclusion rate of paeonol. The calculation methods for paeonol inclusion rate and inclusion complex yield are the same as in Example 1.

[0094] Calculations showed that the paeonol content in the inclusion complex was 2.065%, the inclusion rate of paeonol was 85.4%, and the yield of the inclusion complex was 97.1%.

[0095] Take an appropriate amount of the paeonol inclusion complex described in (2) and store it under high humidity (25℃ / 75%±5%RH), light (illuminance of 4500lx±500lx), and high temperature (60℃) conditions. Samples were taken on the 5th and 10th days under high humidity and light conditions, respectively, and on the 5th, 10th, and 30th days under high temperature conditions, respectively. The content of each sample was determined according to the method under "Content Determination" in Example 1, and the content of paeonol was calculated.

[0096] Calculations showed that the paeonol content in the inclusion complex did not differ significantly within the influencing factor test period (10 days of high humidity, 10 days of light exposure, and 30 days of high temperature), indicating that the paeonol in the inclusion complex prepared by the method of this invention has good stability.

[0097] The experimental results are shown in the table below.

[0098]

Claims

1. A method for in-situ inclusion complex preparation of paeonol in a traditional Chinese medicine compound, characterized in that, Includes the following steps: (1) Take all the medicinal materials in the compound containing peony bark, add 6 to 15 times the amount of water and soak for 0.5 to 2 hours, then extract by steam distillation and collect 1 to 5 times the weight of the medicinal materials in the distillate; (2) Keep the distillate at 25-50°C and stir, and add hydroxypropyl betacyclodextrin to it. The mass ratio of the distillate to hydroxypropyl betacyclodextrin is 5:1 to 50:

1. (3) Continue stirring to dissolve the hydroxypropyl betacyclodextrin, and continue stirring at 25-50°C for 0.5-4 hours to carry out inclusion. After stopping heating, continue stirring until room temperature to obtain the inclusion solution. (4) The inclusion solution was directly freeze-dried to obtain paeonol inclusion complex.

2. The preparation method according to claim 1, characterized in that, In step (1), add 10 times the amount of water and soak for 1 hour, then collect 3 times the weight of the medicinal materials in distillate.

3. The preparation method according to claim 1, characterized in that, In step (2), the mass ratio of the distillate to hydroxypropyl betacyclodextrin is 30:

1.

4. The preparation method according to claim 1, characterized in that, In step (3), the stirring condition under constant temperature is stirring at 30°C for 1 hour.

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