Preparation method and quality control method of radix trichosanthis

Abstract

The invention relates to the technical field of traditional Chinese medicine, and discloses a preparation method and a quality control method of radix trichosanthis. Adding water, grinding into a radix trichosanthis medium powder suspension, and filtering the obtained radix trichosanthis medium powder suspension with a 80-100-mesh screen to obtain a radix trichosanthis filtrate; taking the radix trichosanthis filtrate, putting the radix trichosanthis filtrate into a 200-350-mesh filter bag, suspending until no water drops, adding water, and filtering until the content of residual protein in the filtrate is lower than a control level, so as to obtain a radix trichosanthis filter cake; adding water into the radices trichosanthis filter cake, uniformly stirring, and removing supernate to obtain purified radices trichosanthis; drying the cleaned radix trichosanthis at low temperature to obtain dried radix trichosanthis; scattering, sieving and drying to obtain a radix trichosanthis finished product. By optimizing the steps of grinding, filtering, precipitation purification and temperature-controlled drying, the preparation time of the radix trichosanthis is shortened, the yield is increased, the dispersity of the radix trichosanthis in water is improved, a residual protein mass control method is established, and the preparation process level and the product quality of the radix trichosanthis are improved.

Description

Technical Field

[0001] This application relates to the field of traditional Chinese medicine technology, specifically to the preparation method and quality control method of Trichosanthes kirilowii pollen. Background Technology

[0002] Trichosanthes kirilowii is a traditional Chinese medicine derived from the Cucurbitaceae plant Trichosanthes kirilowii (Trichosanthes chinensis). Trichosanthes kirilowiiMaxim. ) or double-sided Trichosanthes kirilowii ( Trichosanthes rosthornii Harms The dried root of *Trichosanthes kirilowii* (also known as Tianhuafen) was described by Li Shizhen of the Ming Dynasty as having a powder as white as snow. The *Shennong Bencao Jing* (Shennong's Classic of Materia Medica) from the Han Dynasty records that it "treats thirst, fever, restlessness, and high fever." Tianhuafen is sweet and cold in nature, possessing the effects of clearing heat, purging fire, and promoting body fluid production to quench thirst. It has been a key medicine in traditional Chinese medicine for treating thirst since ancient times, and its clinical treatment of diabetes mellitus has shown significant effects. Besides its traditional applications in clearing heat, promoting body fluid production, reducing swelling, and draining pus, Tianhuafen is now also used in modern pharmaceutical research and development, showing potential for lowering blood sugar, anti-tumor, and antiviral effects. It can also be used in health products or foods to lower blood sugar and regulate immunity, demonstrating broad prospects for development and application.

[0003] The traditional method of processing Trichosanthes root powder was created by Sun Simiao, the "King of Medicine" of the Tang Dynasty. The *Qianjin Fang* (Prescriptions Worth a Thousand Gold Pieces) records a "secret recipe for treating severe thirst with Trichosanthes root powder: Dig deep into a large Trichosanthes root, peel off the outer skin down to the white part, cut into 1-inch pieces, soak in water for one day and one night, change the water and soak for five days, then take it out, mash it thoroughly, grind it, filter it through a silk bag, and dry it using the same method as for powder. Take one spoonful with water, three or four times a day." From the Tang and Song Dynasties to the late Ming Dynasty, medical scholars of various dynasties used Trichosanthes root to process into Trichosanthes root powder for medicinal use, treating febrile diseases, yin deficiency and fluid depletion, diabetes, and thirst, with good results. The Trichosanthes root powder used from the Qing Dynasty to the present is simply sliced ​​Trichosanthes root used as medicine; it has the name of powder but not the actual substance, and its processing method is no longer the same.

[0004] Traditionally, Trichosanthes root powder is bitter and cold in nature; excessive or prolonged use may damage the spleen's yang energy and cause diarrhea. The water-soluble protein in Trichosanthes root powder has pharmacological effects that can damage placental villi cells, causing embryonic death, and may also cause skin damage. Current traditional processes use Trichosanthes root sedimentation for powdering, involving prolonged soaking and washing in large amounts of water. This largely removes the water-soluble protein, avoiding adverse reactions. However, prolonged soaking in water makes the medicinal material highly susceptible to microbial growth, leading to rancidity and spoilage. Furthermore, traditional processes lack quality control over residual protein in Trichosanthes root powder and cannot monitor its quantity. Additionally, the traditional method of preparing Trichosanthes root powder is cumbersome, prone to clumping, and inconvenient for patients.

[0005] Therefore, it is necessary to develop a new method for preparing Trichosanthes kirilowii pollen and its quality control method, so as to shorten the soaking time of the medicinal material, increase the yield of the medicinal material during the preparation process, and reduce the content of water-soluble protein in Trichosanthes kirilowii pollen, thereby reducing or avoiding adverse reactions caused by water-soluble protein in Trichosanthes kirilowii pollen after consumption, and improving the dispersion characteristics of Trichosanthes kirilowii pollen in water, simplifying the brewing process before consumption, and facilitating operation and consumption. Summary of the Invention

[0006] The purpose of this application is to overcome the shortcomings of the existing technology and provide a new method for preparing Trichosanthes kirilowii pollen and its quality control method, thereby improving the processing technology of Trichosanthes kirilowii pollen, increasing the yield of medicinal materials, quantitatively controlling the residual amount of water-soluble protein in Trichosanthes kirilowii pollen, and reducing adverse reactions after consumption. It also improves the dispersibility of Trichosanthes kirilowii pollen in water, avoiding the clumping that occurs when brewed with boiling water, which would be inconvenient for consumption.

[0007] To solve the above problems, this application adopts the following technical solution: On the one hand, the specific steps of the preparation method of Trichosanthes kirilowii pollen are as follows: S1: Soak the Trichosanthes kirilowii root in water to obtain fully soaked Trichosanthes kirilowii root; S2: The soaked Trichosanthes kirilowii powder is ground with double the amount of water to form a Trichosanthes kirilowii powder suspension. The obtained Trichosanthes kirilowii powder suspension is filtered through an 80-100 mesh sieve to obtain Trichosanthes kirilowii powder filtrate. S3: Place the pollen filtrate in a 200-350 mesh filter bag, suspend it until no liquid drips, add water again and filter until the content of residual water-soluble protein in the pollen filtrate is lower than the control level, and obtain pollen filter cake. S4: Add water to the powdered trichosanthes filter cake and stir. After standing, remove the supernatant to obtain the purified powdered trichosanthes. S5: The purified precipitate of Trichosanthes kirilowii is dried at low temperature to a moisture content of 35% to 45% to obtain dried Trichosanthes kirilowii; S6: The dried Trichosanthes kirilowii is broken up until it can pass through a 24-mesh sieve, and then dried to a moisture content of less than 10% to obtain the finished Trichosanthes kirilowii.

[0008] Optionally, it may also include a quality control method for pollen, the steps of which include: S7: Soak the finished product of Trichosanthes kirilowii in water, filter it, and take the filtrate, or take the filtrate of Trichosanthes kirilowii during the preparation of Trichosanthes kirilowii; S8: Ninhydrin test solution is added to the filtrate of the finished product of Trichosanthes kirilowii or the filtrate of Trichosanthes kirilowii during the preparation of Trichosanthes kirilowii, heated in a water bath, and the absorbance is measured after cooling. S9: If the absorbance of the sample solution to be tested is greater than or equal to 0.1, then step S3 needs to be repeated; if the absorbance of the sample solution to be tested is less than 0.1, then the residual amount of pollen soluble protein meets the requirements.

[0009] In steps S1-S2, soaking and softening the medicinal materials facilitates the grinding of Trichosanthes kirilowii powder and avoids the generation of a large amount of dust during the grinding process of dry medicinal materials. The term "Trichosanthes kirilowii powder" refers to powder whose particle size can all pass through a 65-mesh sieve, with no more than 60% of the powder passing through an 80-mesh sieve. In step S2, the yield of coarsely ground Tianhua powder is low, and the residual protein content is high; while the impurity content of finely ground Tianhua powder is high. Therefore, grinding into coarse or fine powder is not conducive to producing high-quality Tianhua powder products. The sieve selected is an 80-100 mesh sieve to separate impurities such as wood fibers.

[0010] The water mentioned in the preparation method includes drinking water, purified water, deionized water, and ultrapure water; Preferably, the soaking time for the Trichosanthes kirilowii root is 8 to 12 hours; Preferably, the standing time of the pollen filter cake in step S4 after adding water is 6 to 8 hours; Preferably, in step S1, the mass ratio of the water to the mass of the Trichosanthes kirilowii medicinal material is 8-12:1; Preferably, in step S2, the mass ratio of the water to the soaked pollen is 20-25:1; Preferably, in step S3, the mass ratio of the water to the pollen filtrate is 10-15:1; Preferably, in step S4, the mass ratio of the water to the powder cake is 8 to 12:1.

[0011] Furthermore, in step S3, the filter screen is selected to be suspended in a filter bag for powder making, which can not only efficiently remove impurities and water-soluble components, but also significantly reduce the powder making time.

[0012] Furthermore, in step S4, the purpose of using precipitation purification is to remove lighter tissue debris and other impurities. By combining the filtration and precipitation purification methods of S3 and S4, impurities and water-soluble components can be removed efficiently, and the powdering time can be significantly shortened.

[0013] In step S2, the pollen is ground into medium powder. Combined with the methods in steps S3 and S4, this process synergistically improves the yield of the pollen, reduces the residual protein content, effectively removes impurities, and improves its appearance. The mesh size of the sieve and filter bag used in the comprehensive purification process significantly affects the filtration effect of the pollen. Applying steps S2 and S3 / S4 alone cannot achieve the desired pollen preparation effect as described in this application.

[0014] Furthermore, in step S5, the low-temperature drying range is 45 to 68°C, and the moisture content of the pollen after water control is 35% to 45%, to prevent the pollen from gelatinizing during subsequent processing. During the drying process, the temperature must not exceed 68℃, otherwise the trichosanthes starch will rapidly gelatinize and denature. Below 45℃, the drying time increases significantly, easily causing the trichosanthes starch product to become rancid and spoil.

[0015] If the moisture content of the pollen is below 35% during the drying process, its dispersibility in water cannot be changed after subsequent drying, and it will easily clump together when brewed directly. If the moisture content of the pollen is above 45%, it will easily gelatinize and clump together, making it difficult to disperse.

[0016] Furthermore, in step S6, the drying temperature is 90-100℃, and the moisture content of the pollen is controlled to be <10.0% during the drying process.

[0017] In this process, step S5 utilizes low-temperature drying combined with step S6 high-temperature drying. The synergistic effect of these two methods alters the dispersibility of the powder in water without causing gelatinization or denaturation. This improves upon the traditional method of using powder directly for brewing, which often results in clumping and making the powder easier to brew. Applying steps S5 and S6 alone cannot achieve the powder preparation effect described in this application.

[0018] On the other hand, a method for quality control of Trichosanthes kirilowii pollen is provided for detecting the residual amount of water-soluble protein in Trichosanthes kirilowii pollen; The reaction principle of ninhydrin color development is as follows: ninhydrin reacts with proteins containing free α-amino and α-carboxyl groups to produce a blue-violet substance, which has the highest absorbance at a wavelength of 570 nm. Since pollen contains water-soluble proteins, the color development reaction of ninhydrin can be used to indicate the residual amount of water-soluble proteins in pollen.

[0019] The mass ratio of pollen to water was 1:90 to 110; the volume ratio of the filtrate to ninhydrin was 25-30:1, and the solution was heated in a boiling water bath for 30 minutes.

[0020] Furthermore, the method for preparing *Trichosanthes kirilowii* pollen is used to improve the production process of *Trichosanthes kirilowii* pollen.

[0021] The aforementioned powder drying-baking technology can be applied to improve the brewing and consumption characteristics of powdered products such as powdered trichosanthes.

[0022] Furthermore, the quality control method is applied to the detection of residual protein in pollen to ensure that the residual protein content in pollen is reduced to trace levels, effectively reducing adverse reactions caused by pollen.

[0023] The beneficial effects of this application are: 1. Traditional processing of Trichosanthes kirilowii root takes more than 6 days, while the processing time of the patented Trichosanthes kirilowii root process in this application is only 2 days, which significantly reduces the processing time. In the traditional process of making Trichosanthes kirilowii root, the medicinal material is very prone to rancidity due to microbial growth during the long soaking process, resulting in the adverse consequences of the medicinal material deteriorating. The patented Trichosanthes kirilowii root process in this application significantly shortens the soaking time of the medicinal material, effectively reducing the risk of the medicinal material deteriorating.

[0024] 2. The traditional process for producing Trichosanthes kirilowii root extract yields approximately 35%. The method described in this application, through the synergistic application of filtration and precipitation purification methods using sieves with specific mesh sizes, increases the product yield to approximately 55%, significantly improving the yield of the medicinal material. It also effectively removes impurities such as water-soluble proteins, resulting in Trichosanthes kirilowii root extract with a "snow-white" appearance.

[0025] 3. Traditionally processed Trichosanthes kirilowii pollen does not undergo residual control of water-soluble proteins, posing a risk of adverse reactions after consumption. This application also provides a method for testing residual proteins in Trichosanthes kirilowii pollen. Applying this method during the manufacturing process can effectively monitor the residual protein content, ensuring it is reduced to trace levels, thus effectively reducing the risk of adverse reactions. This application can not only detect the residual amount of water-soluble proteins in Trichosanthes kirilowii pollen during the preparation of finished products, but also detect whether the residual amount of water-soluble proteins in finished Trichosanthes kirilowii pollen products prepared using the method of this application and other finished Trichosanthes kirilowii pollen products meets the standards.

[0026] 4. Traditionally processed Trichosanthes kirilowii root powder tends to clump when directly brewed. It requires dilution with a small amount of room temperature water before adding boiling water, a cumbersome process that reduces patient compliance. The Trichosanthes kirilowii root powder preparation method described in this application improves brewing characteristics through a combination of low-temperature drying and high-temperature baking. It can be brewed directly with warm or boiling water without clumping, making it easier for patients to take and significantly improving patient compliance.

[0027] 5. This application utilizes a rational method for preparing Trichosanthes kirilowii pollen, removing water-soluble proteins and significantly reducing the risk of adverse reactions. Compared to the processed Trichosanthes kirilowii pollen slices in the current Chinese Pharmacopoeia, the Trichosanthes kirilowii pollen prepared in this application does not require decoction and can be directly brewed and consumed. It also does not clump together, making it convenient to take.

[0028] 6. The Trichosanthes kirilowii pollen prepared according to this application has significant effects on stabilizing blood sugar levels and relieving thirst symptoms in diabetic patients. At the same time, it is simple and convenient to brew directly, which helps to increase patient compliance and has broad market potential and economic value. Attached Figure Description

[0029] Figure 1The images are 100x magnification images of Trichosanthes kirilowii cells under an optical microscope under different grinding degrees and purification methods (Figure A is Trichosanthes kirilowii medium powder purified by filtration and precipitation in Example 1; Figure B is Trichosanthes kirilowii coarse powder purified by filtration and precipitation in Comparative Example 1; Figure C is Trichosanthes kirilowii fine powder purified by filtration and precipitation in Comparative Example 2; Figure D is Trichosanthes kirilowii medium powder purified by water washing, precipitation, and gauze filtration in Comparative Example 6).

[0030] Figure 2 Comparison charts showing the properties and brewing characteristics of Trichosanthes kirilowii pollen products after varying drying temperatures and times (Figure A: Drying temperature of 60℃ and drying time of 40 min in Example 1; Figure B: Drying temperature of 55℃ and drying time of 40 min in Example 3; Figure C: Drying temperature of 68℃ and drying time of 10 min in Comparative Example 7; Figure D: Drying temperature of 45℃ and drying time of 120 min in Comparative Example 8; Figure E: Results of brewing Trichosanthes kirilowii pollen in Example 1, with no clumping; Figure F: Drying temperature of 60℃ and drying time of 50 min in Example 4; Figure G: Drying temperature of 60℃ and drying time of 30 min in Comparative Example 9; Figure H: Drying temperature of 60℃ and drying time of 60 min in Comparative Example 10).

[0031] Figure 3 Comparison of the properties of pollen products with different drying temperatures and times (Figure A is the drying temperature of 90℃ and drying time of 120 min in Example 1; Figure B is the drying temperature of 100℃ and drying time of 120 min in Example 3; Figure C is the drying temperature of 90℃ and drying time of 130 min in Example 6).

[0032] Figure 4 shows a comparison of the appearance of the powder prepared by the method of this application and the traditional process (Figure A shows the powder prepared by this application; Figure B shows the powder prepared by the traditional process).

[0033] Figure 5 The effect of different amounts of ninhydrin added on the colorimetric reaction in this application is shown in the figure (from left to right, the amounts of ninhydrin added are 0.1 mL, 0.2 mL, 0.3 mL, 0.5 mL, and 1 mL, respectively).

[0034] Figure 6 Color changes caused by different heating times (heating times from left to right are 40 min, 30 min, and 20 min).

[0035] Figure 7 The absorbance values ​​are the average values ​​of different batches of finished Tianhuafen (a type of medicinal powder). Detailed Implementation

[0036] The technical solution of this application is described in further detail below with reference to figures and text, but the scope of protection of this application is not limited to what is described below.

[0037] Example 1 The steps of a method for preparing pollen from Trichosanthes kirilowii are as follows: S1: Add 10 times the weight of the Trichosanthes kirilowii root to water and soak the root for 10 hours to obtain fully soaked Trichosanthes kirilowii root. S2: Add 20 times the amount of water to the soaked Tianhua powder, grind it into medium powder using a universal pulverizer, filter the obtained Tianhua powder medium powder through an 80-mesh sieve to remove impurities such as wood fibers, and obtain Tianhua powder filtrate. S3: Take the pollen filtrate, place it in a 300-mesh filter bag, and suspend it until it stops dripping water to obtain a filter cake. Add 10 times the amount of water to the filter cake and repeat the above steps until the residual protein content in the filtrate is lower than the control level to obtain a pollen filter cake. S4: Take the above-mentioned Trichosanthes root filter cake, add 10 times the amount of water and stir well. Let it stand for 6 hours, pour off the supernatant, remove tissue debris and other impurities, and obtain the Trichosanthes root starch after precipitation and purification. S5: Take the precipitated and purified Trichosanthes starch, dry it at a low temperature of 60°C for 40 minutes until the moisture content is 35% to 45%, and obtain dried Trichosanthes. S6: The dried Tianhua powder is broken up until it can pass through a 24-mesh sieve, and then dried at 90°C for 120 minutes until the moisture content is <10.0%, thus obtaining the finished Tianhua powder product.

[0038] Comparative Example 1 The difference between the method for preparing Trichosanthes kirilowii pollen and Example 1 is that the soaked Trichosanthes kirilowii pollen described in S2 is ground into coarse powder under controlled equipment, and the yield, residual protein content, and amount of impurities in the Trichosanthes kirilowii pollen under this method are measured.

[0039] Comparative Example 2 The difference between the method for preparing Trichosanthes kirilowii pollen and Example 1 is that, in S2, the thoroughly soaked Trichosanthes kirilowii pollen is ground into fine powder under controlled equipment, and the yield, residual protein content, and amount of impurities in the Trichosanthes kirilowii pollen are measured under a microscope.

[0040] By comparing the yield, protein residue, and impurities of Trichosanthes kirilowii pollen, the quality of Trichosanthes kirilowii pollen prepared in Example 1 and Comparative Examples 1-2 under different grinding degrees was observed, as shown in Table 1.

[0041] Table 1. Pollen quality under different grinding degrees Table 1 shows that grinding Trichosanthes kirilowii pollen into medium-sized powder increased the product yield by 18.7% compared to coarse powder, and reduced the residual protein content by 15 times. While fine powder had a higher product yield and similar residual protein content compared to fine powder, it showed significantly more impurities observed under a microscope. Therefore, grinding Trichosanthes kirilowii pollen into medium-sized powder helps improve the yield and reduce residual protein and impurities.

[0042] Example 2 The difference between the preparation method of Trichosanthes kirilowii pollen and Example 1 is that the sieve used in S2 is 100 mesh.

[0043] Comparative Example 3 The difference between the preparation method of Trichosanthes kirilowii pollen and Example 1 is that the sieve used in S2 is 120 mesh.

[0044] Comparative Example 4 The difference between the method for preparing Trichosanthes kirilowii pollen and Example 1 is that the filter bag in S3 has a mesh size of 200.

[0045] Comparative Example 5 The difference between the method for preparing pollen from Trichosanthes kirilowii and Example 1 is that the filter bag described in S3 has a mesh size of 400.

[0046] By comparing the number of filtrations, the yield of Trichosanthes kirilowii pollen, the amount of residual protein, and the content of impurities in Trichosanthes kirilowii pollen, the process efficiency and quality of Trichosanthes kirilowii pollen preparation in Examples 1 and Comparative Examples 3 to 5 under different sieve mesh sizes and filter bag mesh sizes are observed, as shown in Table 2.

[0047] Table 2. Pollen quality after sieving with different screen mesh sizes and filter bag pore sizes. Table 2 shows that in Example 2, when the pollen was filtered using a 100-mesh sieve, the final product yield decreased by about 5% compared to Example 1, which is within an acceptable range. In Comparative Example 3, when the pollen was filtered using a 120-mesh sieve, the yield of the final product decreased significantly due to the smaller sieve aperture, even after three filtrations. Therefore, the suitable sieve mesh size is between 80 and 100 mesh.

[0048] Comparative Example 4 used a 200-mesh filter bag with a larger pore size than that specified in this application, achieving a similar product yield and residual protein content as Example 1. It required six filtrations, significantly increasing the filtration time and the risk of product rancidity. Comparative Example 5 used a 400-mesh filter bag with a smaller pore size than that specified in this application; as a result, the mixture of filtrate and water could not be separated, and the filtration purification process could not be completed. Therefore, the suitable mesh size for filter bags is between 200 and 350 mesh.

[0049] Comparative Example 6 The method for preparing Trichosanthes kirilowii pollen differs from that in Example 1 in that the filtration and precipitation in steps S3 and S4 are replaced with traditional gauze filtration and water washing for clarification.

[0050] By comparing the preparation time, the yield of Trichosanthes kirilowii pollen, the amount of residual protein, and the content of impurities in Trichosanthes kirilowii pollen, the process efficiency and quality of Trichosanthes kirilowii pollen under the preparation conditions of Example 1 and Comparative Example 6 are observed, as shown in Table 3.

[0051] Table 3. Pollen under different filtration conditions Table 3 shows that, through Comparative Example 6, it was found that in the preparation of *Trichosanthes kirilowii* pollen, the traditional method of washing, settling, and filtering with gauze resulted in a preparation time of up to 6 days. The product yield was significantly lower than in Example 1, and the pH of the product solution was acidic, indicating that rancidity occurred due to the excessive preparation time. In contrast, Comparative Example 7, which simply ground *Trichosanthes kirilowii* pollen into a medium powder and filtered it through a 300-mesh filter bag without precipitation, showed more tissue debris under a microscope, indicating that it contained more impurities than in Example 1. Microscopic images of starch granules and impurities in *Trichosanthes kirilowii* pollen prepared with different grinding degrees and purification processes are shown below. Figure 1 Figure A shows cells that are uniform in size, evenly distributed, plump, numerous, and without clumping; Figure B shows cells that are uneven in size, severely clumped, unevenly distributed, not plump, and few in number; Figure C shows cells that are small, uneven in size, not severely clumped, and unevenly distributed; Figure D shows cells that are of moderate size, relatively uniform in size, not severely clumped, but few in number.

[0052] As can be seen from the above, the combined method of filtration and precipitation purification can effectively shorten the preparation time of Trichosanthes kirilowii pollen, reduce the possibility of rancidity, and effectively remove impurities. Implementing steps S3 and S4 alone cannot achieve the Trichosanthes kirilowii pollen preparation effect described in this application.

[0053] Example 3 The difference between the preparation method of Trichosanthes kirilowii pollen and that in Example 1 is that the low-temperature drying temperature of Trichosanthes kirilowii pollen in S5 is 55°C.

[0054] Comparative Example 7 The difference between the preparation method of Trichosanthes kirilowii pollen and that in Example 1 is that the drying temperature of Trichosanthes kirilowii pollen in S5 is 68°C.

[0055] Comparative Example 8 The difference between the preparation method of Trichosanthes kirilowii pollen and that in Example 1 is that the drying temperature of Trichosanthes kirilowii pollen in S5 is 45°C.

[0056] Example 4 The difference between the preparation method of Trichosanthes kirilowii pollen and Example 1 is that the drying time of Trichosanthes kirilowii pollen in S5 is increased to 50 minutes.

[0057] Comparative Example 9 The preparation method of Trichosanthes kirilowii pollen differs from that in Example 1 in that the drying time of Trichosanthes kirilowii pollen in S5 is shortened to 30 minutes, resulting in a water content of Trichosanthes kirilowii pollen that is higher than the upper limit of 45% water content in this application.

[0058] Comparative Example 10 The preparation method of Trichosanthes kirilowii pollen differs from that in Example 1 in that the drying time of Trichosanthes kirilowii pollen in S5 is extended to 60 minutes, so that the moisture content of Trichosanthes kirilowii pollen is lower than the lower limit of 35% of the moisture content of this application.

[0059] Example 5 The difference between the preparation method of Trichosanthes kirilowii pollen and that in Example 1 is that the drying temperature of Trichosanthes kirilowii pollen in S6 is 100°C.

[0060] Example 6 The preparation method of Trichosanthes kirilowii pollen differs from that in Example 1 in that the drying time of Trichosanthes kirilowii pollen in S6 is extended to 130 minutes.

[0061] The moisture content of the pollen after moisture control was determined by adjusting the drying temperature and drying time; the moisture content of the finished pollen product was determined by adjusting the drying temperature and drying time. The degree of gelatinization, dispersibility in water, and properties of the finished pollen products in Examples 3-6 and Comparative Examples 7-10 were observed in subsequent processes, as shown in Table 4.

[0062] Table 4. Effects of drying and baking temperature and time on pollen. The moisture content of dried *Trichosanthes kirilowii* pollen was measured at different drying temperatures and times. Table 4 shows that in Example 3, the temperature was lowered to 55℃, and the moisture content was measured to be 44.3%. After subsequent high-temperature drying, the *Trichosanthes kirilowii* pollen showed no gelatinization and did not clump together. In Comparative Example 7, the drying temperature was increased to 68℃. After only 10 minutes of drying, the *Trichosanthes kirilowii* pollen rapidly gelatinized and solidified into white lumps; therefore, the drying temperature should not exceed 68℃. In Comparative Example 8, the drying temperature was lowered to 45℃, and after 120 minutes of drying, the moisture content was controlled to approximately 40%. The significantly increased drying time increased energy consumption and the possibility of rancidity and deterioration of the *Trichosanthes kirilowii* pollen in a humid environment. Therefore, the drying temperature should not be lower than 45℃. In Example 4, the drying time was increased to 50 minutes, and the moisture content was measured to be 36.5%. After subsequent high-temperature drying, the *Trichosanthes kirilowii* pollen showed no gelatinization and did not clump together. Comparative Example 9 shortened the drying time to 30 minutes, and the moisture content was measured to be 58.7%. After subsequent high-temperature drying, the *Trichosanthes kirilowii* powder showed gelatinization, coagulating into brown hard lumps, indicating that the moisture content after drying exceeded 45%, making the *Trichosanthes kirilowii* powder prone to gelatinization and denaturation. Comparative Example 10 increased the drying time to 60 minutes, and the moisture content was measured to be 28.3%. After subsequent high-temperature drying, the *Trichosanthes kirilowii* powder did not show gelatinization. However, when directly brewed with boiling water, clearly visible, non-dispersible clumps appeared in the solution, indicating that a moisture content below 35% could not improve the dispersibility of the *Trichosanthes kirilowii* powder in water. The properties of the finished *Trichosanthes kirilowii* powder under different drying conditions are as follows: Figure 2 As shown, Figure 2 The pollen from A, B, and F, dried at 55-65℃ for 40-50 minutes, is white and fine, and is easily diluted with warm water and brewed with boiling water. Figure 2 E); When the drying temperature of the pollen exceeded 65℃, severe clumping and browning occurred. Figure 2 C) When the drying temperature is below 55℃, pollen is prone to clumping; when the drying time of pollen is less than 40 min, clumping and browning are likely to occur; when the drying time of pollen is greater than 50 min, it becomes difficult to disperse.

[0063] Therefore, observations from Examples 3-4 and Comparative Examples 7-10 revealed that controlling the drying temperature of Trichosanthes kirilowii pollen at 55-65°C and the drying time at 40-50 minutes resulted in a moisture content of 35%-45% after drying, leading to the optimal quality of the prepared Trichosanthes kirilowii pollen.

[0064] The moisture content of the dried *Trichosanthes kirilowii* powder was determined under different baking temperatures and times. Table 4 shows that in Example 5, the drying temperature was 100℃, and after 120 minutes, the moisture content of the finished *Trichosanthes kirilowii* powder was 4.5%. The finished product was white and did not clump when brewed. In Example 5, the drying temperature was 100℃, and the drying time was increased to 130 minutes. The moisture content of the finished *Trichosanthes kirilowii* powder was 2.1%. The finished product was a slightly yellowish white powder and did not clump when brewed. The properties of the finished *Trichosanthes kirilowii* powder under different drying conditions are as follows: Figure 3 As shown.

[0065] Therefore, observations from Examples 5-6 show that the suitable drying temperature for Trichosanthes kirilowii pollen is controlled at 90-100℃ and the drying time is 120-130 minutes, so that the moisture content of the dried Trichosanthes kirilowii pollen is controlled below 10%, and the final product of Trichosanthes kirilowii pollen can achieve good quality, with a white color, good dispersibility in water, and will not clump together when brewed.

[0066] As shown above, in the preparation method of Trichosanthes kirilowii root powder, the drying temperature is 45-68℃, and the moisture content is controlled at 35%-45%; the drying temperature is 90-100℃, and the moisture content is controlled at <10%. Utilizing a combination of low-temperature drying and high-temperature drying, the two synergistically alter the dispersibility of Trichosanthes kirilowii root powder in water without gelatinizing or denaturing it. This improves upon the traditional method of directly brewing Trichosanthes kirilowii root powder, which easily clumps together, thus achieving the goal of easy brewing. Applying steps S5 and S6 alone cannot achieve the Trichosanthes kirilowii root powder preparation effect of this application.

[0067] Comparative Example 11 The traditional preparation process of Trichosanthes kirilowii pollen includes the following steps: 1) Take the medicinal material Trichosanthes kirilowii, cut it into 1-inch sections, soak it in water, change the water once a day, and soak for 3 days; 2) Take the soaked Trichosanthes kirilowii root, grind it, filter it with gauze, and let the filtrate settle and change the water every 3 days; 3) Take the precipitated Trichosanthes kirilowii pollen and dry it.

[0068] By comparing the traditional preparation process of Trichosanthes kirilowii pollen with the preparation method of this application, under the indicators of preparation time, residual protein content of Trichosanthes kirilowii pollen, water content of Trichosanthes kirilowii pollen, product yield, pH value of product solution, impurity content of Trichosanthes kirilowii pollen, reconstitution properties of Trichosanthes kirilowii pollen, and appearance of Trichosanthes kirilowii pollen (including color, size, and uniformity of Trichosanthes kirilowii pollen), the different performances of the different methods of preparing Trichosanthes kirilowii pollen in Example 1 and Comparative Example 11 in terms of preparation efficiency and product quality were observed. The specific results are shown in Table 5. Table 5. Differences between this application and traditional Tianhua pollen in terms of preparation time and product properties. Table 5 shows that, comparing the different indicators of the traditional and present invention's methods for preparing Trichosanthes kirilowii powder, the present invention, through a comprehensive method combining controlled grinding of the powder with filtration and precipitation, not only improves the yield of Trichosanthes kirilowii powder but also shortens the preparation time, reduces the possibility of product rancidity and deterioration, and ensures that the final product has a neutral pH, which is beneficial for long-term storage. It also thoroughly removes residual protein and impurities from the powder. Furthermore, the present invention rationally controls the temperature and time during drying and baking, ensuring that the moisture content of the powder is <10%. The final product is snow-white with uniform powder particles. Simultaneously, the synergistic effect of low-temperature drying and baking improves the dispersibility of the powder in water, allowing for direct brewing with warm or boiling water, making it convenient to consume and improving patient compliance. Figure 4 compares the appearance of the powder prepared by the present invention with that prepared by the traditional method; the powder prepared by the present invention is white and fine (…). Figure 4A The pollen prepared using traditional methods is white lumps or granular powder. Figure 4B ).

[0069] Example 7 The quality control method for pollen (Trichosanthes kirilowii) and the specific steps are as follows: S6: Take 1 g of Trichosanthes kirilowii pollen, add 100 mL of water and soak for 1 h, take 5 mL of the filtrate, add 0.2 mL of ninhydrin, and heat in boiling water for 30 min; S7: Cool the test liquid and measure the absorbance under ultraviolet spectrophotometry at a wavelength of 570 nm.

[0070] Comparative Example 12 The quality control method for Trichosanthes kirilowii pollen differs from that in Example 7 in that (1) the amount of water added is 10 mL.

[0071] Comparative Example 13 The quality control method for Trichosanthes kirilowii pollen differs from that in Example 7 in that (1) the amount of water added is 50 mL.

[0072] Comparative Example 14 The difference between the pollen quality control method in Example 7 and Example 8 is that the amount of water added in (1) is 200 mL.

[0073] Comparative Example 15 The quality control method for Trichosanthes kirilowii pollen differs from that in Example 7 in that (1) the amount of ninhydrin added is 0.1 mL.

[0074] Comparative Example 16 The quality control method for Trichosanthes kirilowii pollen differs from that in Example 7 in that (1) the amount of ninhydrin added is 0.3 mL.

[0075] Comparative Example 17 The quality control method for Trichosanthes kirilowii pollen differs from that in Example 7 in that (1) the amount of ninhydrin added is 0.5 mL.

[0076] Comparative Example 18 The quality control method for Trichosanthes kirilowii pollen differs from that in Example 7 in that (1) the amount of ninhydrin added is 1.0 mL.

[0077] By comparing absorbance values ​​and sample color changes ( Figure 5 The optimal experimental conditions were determined by comparing different amounts of water added and different amounts of ninhydrin added in Examples 7 and 12-18. The specific results are shown in Table 6. Table 6. Absorbance changes of samples with different amounts of ninhydrin. According to Beer-Lambert's law (A = εcl), the absorbance A value measured by UV-Vis spectrophotometry in the range of 0.2-0.7 has the highest confidence level (the linear relationship between absorbance and concentration is usually the most stable, resulting in more accurate measurements). Table 6 shows that 100 mL of water was optimal for sample addition. To minimize the interference of ninhydrin reagent on the experimental data, the amount of ninhydrin reagent used should be reduced as much as possible while ensuring sufficient color development; therefore, the amount of ninhydrin reagent added was determined to be 0.2 mL.

[0078] Comparative Example 19 The quality control method for Trichosanthes kirilowii pollen differs from that in Example 7 in that (1) the boiling water heating time is 20 min.

[0079] Comparative Example 20 The quality control method for Trichosanthes kirilowii pollen differs from that in Example 7 in that (1) the boiling water heating time is 40 min.

[0080] By comparing the color changes of the samples ( Figure 6 The color changes of the samples in Example 7 and Comparative Examples 19-20 were compared after boiling water bath time. The results showed that the color change of the samples was most significant after boiling water heating for 30 min.

[0081] Application Example 1 Seven batches of Trichosanthes kirilowii root slices were collected, with batch numbers 20230211, 20230212, 20230315, 20230326, 20230516, 20230619, and 20231011. The absorbance of the different Trichosanthes kirilowii root slices was measured according to the experimental method in Example 7. Three groups were measured for each batch, and each group was measured twice in parallel, with the average value taken. The results are shown in Table 7.

[0082] Table 7. Absorbance values ​​of different batches of products measured using this application. As shown in Table 7 and Figure 7 As shown, the absorbance of seven batches of Trichosanthes kirilowii root slices was measured. The absorbance values ​​ranged from 0.002 to 0.036, with an average of 0.019 and an arithmetic mean of 0.012. Based on the sensitivity of the UV-Vis spectrophotometer, the required indicator for residual protein in Trichosanthes kirilowii root slices was determined to be an absorbance not exceeding 0.1. For Trichosanthes kirilowii root slices prepared according to this standard, if the absorbance value does not exceed 0.1 when tested for residual protein using this method, the residual protein content in the Trichosanthes kirilowii root slices is considered to meet the standard. The quality control method described in this application can also be used to detect residual protein during the preparation of the Trichosanthes kirilowii root slices. The quality control method for Trichosanthes kirilowii root slices in this application helps simplify protein detection methods, quantitatively control the residual amount of soluble protein in Trichosanthes kirilowii root slices, and reduce adverse reactions after patient consumption.

[0083] The above description is merely a preferred embodiment of the present invention. It should be understood that the present invention is not limited to the forms disclosed herein and should not be construed as excluding other embodiments. It can be used in various other combinations, modifications, and environments, and can be altered within the scope of the concept described herein through the above teachings or related technologies or knowledge. Modifications and variations made by those skilled in the art that do not depart from the spirit and scope of the present invention should be within the protection scope of the appended claims.

Claims

1. A method for preparing a finished product of Trichosanthes kirilowii, characterized in that, Includes the following steps: S1: Soak the Trichosanthes kirilowii root in water to obtain fully soaked Trichosanthes kirilowii root; S2: The soaked Trichosanthes kirilowii pollen is ground with water to form a Trichosanthes kirilowii pollen suspension. The obtained Trichosanthes kirilowii pollen suspension is filtered through an 80-100 mesh sieve to obtain Trichosanthes kirilowii pollen filtrate. S3: Place the pollen filtrate in a 200-350 mesh filter bag, suspend it until no liquid drips, add water again and filter until the content of residual water-soluble protein in the pollen filtrate is lower than the control level, and obtain pollen filter cake. S4: Add water to the powdered trichosanthes filter cake and stir. After standing, remove the supernatant to obtain the purified powdered trichosanthes. S5: The purified Trichosanthes kirilowii powder is dried at low temperature to a moisture content of 35% to 45% to obtain dried Trichosanthes kirilowii powder. S6: The dried Tianhua powder is broken up until it can pass through a 24-mesh sieve, and then dried until the moisture content is less than 10%, thus obtaining the finished Tianhua powder product.

2. The preparation method according to claim 1, characterized in that: In step S1, the soaking time is 8 to 12 hours; In step S4, the pollen filter cake is left to stand for 6 to 8 hours after being added to water.

3. The preparation method according to claim 1, characterized in that: In step S1, the mass ratio of the water to the mass of the *Trichosanthes kirilowii* medicinal material is 8 to 12:

1. In step S2, the mass ratio of the water to the soaked pollen is 20 to 25:1; In step S3, the mass ratio of the water to the pollen filtrate is 10 to 15:1; In step S4, the mass ratio of the water to the powder cake is 8 to 12:

1.

4. The preparation method according to claim 1, characterized in that: In step S5, the temperature for low-temperature drying is 45–68°C.

5. The preparation method according to claim 1, characterized in that: In step S6, the drying temperature is 90-100°C.

6. A method for quality control of pollen, characterized in that: Includes the following steps: S7: The *Trichosanthes kirilowii* powder product prepared by the method of any one of claims 1 to 4, or the *Trichosanthes kirilowii* powder filter cake obtained by the method of any one of claims 1 to 4 during the preparation of the *Trichosanthes kirilowii* powder product, is soaked in water, filtered, and the filtrate is collected to obtain the sample solution to be tested. S8: Add ninhydrin reagent to the sample solution to be tested, heat in a water bath and cool, and then measure the absorbance; S9: If the absorbance of the sample solution to be tested is greater than or equal to 0.1, then step S3 needs to be repeated; if the absorbance of the sample solution to be tested is less than 0.1, then the residual amount of pollen soluble protein meets the requirements.

7. The quality control method according to claim 6, characterized in that: The mass ratio of the finished product of Trichosanthes kirilowii to water is 1:90 to 110; the volume ratio of the filtrate of the finished product of Trichosanthes kirilowii to ninhydrin is 25-30:1, and it is heated in a boiling water bath for 30 min; the method for determining absorbance includes ultraviolet-visible spectrophotometry, with a detection wavelength of 570 nm.

8. The quality control method according to any one of claims 6 to 7 is used for the detection of residual protein during the preparation of the pollen product.

9. The quality control method as described in any one of claims 6 to 7 is used for quality testing of finished product *Trichosanthes kirilowii* (a type of pollen).

10. The finished product of the Trichosanthes kirilowii pollen prepared by the preparation method according to any one of claims 1 to 5 and the quality control method according to any one of claims 6 to 7.

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