Preparation process of lyophilized traditional Chinese medicine decoction pieces of haemadipsa integrifascialis

Abstract

This invention discloses a preparation process for freeze-dried leech powder, comprising the following steps: taking fresh whole leeches, washing, pre-freezing, homogenizing, freeze-drying, pulverizing, sieving, and sterilizing to obtain the final product. This invention improves the pre-freezing and homogenizing processes, and removes the fishy smell from both the fresh leeches and the freeze-dried leech powder in two separate steps. This avoids changes in the properties and oxidation of the leech material during processing, comprehensively preserving the antithrombin activity of the leech. It can produce freeze-dried leech powder with high activity and low fishy smell without introducing any excipients or additives.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine processing technology, specifically relating to a preparation process of freeze-dried Hirudo medicinalis powder. Background Technology

[0002] Freeze-dried leech powder is derived from the leech *Hirudo medicinalis*, an animal belonging to the family Hirudinidae. Poecilobdella manillensis Lesson.'s processed whole fresh farmed leeches (Yunnan Province Standard for Traditional Chinese Medicine Slices, YPBZ-0199-2013). The leech *Hirudo medicinalis* is one of the larger species of common blood-sucking leeches, feeding on the blood of humans and livestock, and living in paddy fields, ditches, or ponds. Modern research shows that *Hirudo medicinalis* contains hirudin, amino acids, fatty acids, trace elements, and other components, possessing pharmacological effects such as anticoagulation, platelet aggregation inhibition, antithrombosis, lipid-lowering, and anti-tumor properties. Clinically, it is widely used to treat cerebral thrombosis, cerebral hemorrhage, hyperviscosity syndrome, hyperlipidemia, hypertension, and coronary heart disease.

[0003] The preparation process of freeze-dried leech powder generally includes: taking fresh whole leeches, washing them, adding an appropriate amount of water, pre-freezing, homogenizing, freeze-drying, pulverizing, passing through a No. 3 sieve, and sterilizing. However, the pre-freezing of leeches forms lumps, making it difficult to disperse during homogenization and easily causing mechanical damage to the leeches (homogenization speed is generally tens of thousands of rpm), reducing the activity of the finished freeze-dried leech powder.

[0004] Furthermore, the freeze-dried leech powder uses raw leeches, giving it a strong fishy odor. Traditional chemical deodorization methods (such as adding acids, alkalis, salts, flavoring agents, and other organic reagents) require the introduction of chemical reagents, affecting the purity and medicinal properties of the herbal medicine and easily leaving residues. Biological deodorization methods (such as yeast fermentation) use virtually no organic reagents and are recognized as a green method with good development prospects. During the metabolism of yeast, aldehydes and alcohols with specific odors are metabolized into acids, thereby achieving the purpose of removing the odor. However, this is often accompanied by the special off-odor of yeast fermentation; in addition, the metabolism of microorganisms consumes some nutrients, thus causing the loss of active ingredients in the leech.

[0005] Cyclodextrin encapsulation, a physical method for removing fishy odors, is effective; however, the excipients introduced are unsuitable for the production of traditional Chinese medicine decoction pieces. Adsorption is a green and safe method, as the adsorbent material can completely remove the odor without any residue or odor. However, existing adsorption methods (such as activated carbon adsorption) are not very effective at removing fishy odors and easily absorb the active ingredients of leeches.

[0006] In summary, how to provide a preparation process for freeze-dried leech powder, and how to comprehensively improve the quality and deodorizing effect of freeze-dried leech powder by optimizing the processing technology and deodorizing methods, is an urgent problem to be solved. Summary of the Invention

[0007] The technical problem to be solved by the present invention is to overcome the shortcomings of the prior art and provide a preparation process for freeze-dried Hirudo medicinal materials. By improving the pre-freezing and homogenization processes, and by removing the fishy smell from fresh Hirudo medicinal materials and freeze-dried Hirudo medicinal materials in two separate steps, the changes and oxidation of Hirudo medicinal materials caused by the processing process can be avoided. The antithrombin activity of Hirudo medicinal materials is comprehensively preserved. Freeze-dried Hirudo medicinal materials with high activity and low fishy smell can be obtained without introducing any excipients or additives.

[0008] The first objective of this invention is to provide a preparation process for freeze-dried Hirudo medicinalis powder. Because the preparation of freeze-dried Hirudo medicinalis powder requires pre-freezing, homogenization, and then freeze-drying, the pre-freezing process results in a blocky frozen material that is difficult to disperse during homogenization. This leads to a longer homogenization time, which can easily oxidize the medicinal material or increase shear force, potentially causing mechanical damage. Therefore, this invention improves the pre-freezing and homogenization processes, enabling the acquisition of well-frozen and uniformly dispersed Hirudo medicinalis before freeze-drying without damaging the medicinal material.

[0009] To achieve the first objective, the present invention adopts the following technical solution: A process for preparing freeze-dried leech powder, a traditional Chinese medicine, includes the following steps: taking fresh whole leeches, washing them, pre-freezing them, homogenizing them, freeze-drying them, pulverizing them, passing them through a No. 3 sieve (40-60 mesh), and sterilizing them to obtain the final product.

[0010] Preferably, pre-freezing and homogenization specifically include: (1) Add water to the cleaned leeches at a mass percentage of 1-2%, mix evenly, and place at -(5-10)℃ for the first freezing. When the material hardness is 4000-7000g, stop freezing and perform manual homogenization (stir repeatedly with a stainless steel grinding rod) for 3-5 minutes. (2) Add water to the material obtained in step (1) by 3-4% by mass, mix evenly and place at -(10-20)℃ for a second freezing. When the material hardness is 10000-15000g, stop freezing and perform mechanical homogenization at a speed of 3000-10000r / min for 1-3min.

[0011] Preferably, in step (1), the material is frozen until the hardness is 5000-6000g and the homogenization time is 3min.

[0012] Preferably, in step (2), the material is frozen to a hardness of 13000-14000g, the homogenization speed is 5000-8000r / min, and the homogenization time is 1min.

[0013] Preferably, the freeze-drying temperature is -45-50℃, the time is 23-26h, and the vacuum degree is below 20Pa.

[0014] The second objective of this invention is to remove the fishy smell from fresh leeches and freeze-dried leech powder in two separate steps, effectively reducing the fishy smell of freeze-dried leech powder while preserving the activity of the medicinal slices and improving their palatability.

[0015] To achieve the second objective, the present invention adopts the following technical solution: A method for removing fishy odor in the preparation process of freeze-dried leech powder, the method comprising a first deodorization treatment and a second deodorization treatment.

[0016] The first deodorization treatment involves removing the fishy smell from fresh leeches. The specific steps include: after washing the leeches, adding water and starch, mixing evenly, then adding deoxyarbutin and fructose, soaking for 20-50 minutes, and then rinsing with clean water.

[0017] The second deodorization process involves removing the odor from the freeze-dried and pulverized leech powder. Specific steps include: soaking activated carbon in a sodium hydroxide solution of 4-6 times its weight, then removing and drying it; soaking it in an organic acid solution, then evaporating it to dryness to obtain modified activated carbon; spreading the freeze-dried and pulverized leech powder evenly on a sieve; then spreading the modified activated carbon evenly on the sieve and fixing it directly above the sieve containing the leech powder; and finally, blowing air from below the sieve containing the leech powder.

[0018] Preferably, in the first deodorization treatment, the mass ratio of water, starch, deoxyarbutin, and fructose is 1:(0.05-0.1):(0.005-0.02):(0.02-0.04), and the amount of water added is 3-6 times the mass of the washed leeches.

[0019] Preferably, in the second deodorization treatment, the activated carbon is 100-180 mesh, the mass concentration of the sodium hydroxide solution is 1-2%, the soaking time of the sodium hydroxide solution is 1-2 hours, and the soaking time of the organic acid solution is 40-80 minutes.

[0020] Preferably, the pulverized Hirudo medicinalis powder is spread on a sieve of 80-120 mesh, and the modified activated carbon is spread on a sieve of 200-300 mesh, with the mass of the modified activated carbon on the sieve being 10-100% of the mass of the Hirudo medicinalis powder. More preferably, the mass of the modified activated carbon on the sieve is 30-100% of the mass of the Hirudo medicinalis powder. Preferably, the blowing conditions include: room temperature, wind speed of 5-10 m / s, and blowing time of 20-40 min.

[0021] Preferably, in the second deodorizing treatment, the organic acid is veratril acid, acetic acid, citric acid, vanillic acid, malic acid, or hawthorn acid. More preferably, the organic acid is veratril acid.

[0022] Preferably, in the second deodorization treatment, the organic acid in the organic acid solution accounts for 3-6% of the total mass, and the solvent in the organic acid solution is one or more of water, ethanol, diethyl ether, and acetone. The amount of organic acid solution added is 5-10 times the mass of the activated carbon after soaking and drying in sodium hydroxide solution.

[0023] Technical effects of the present invention: 1. This invention improves the pre-freezing and homogenization process. First, the leeches are partially frozen (to prevent them from forming lumps of frozen material), then they are manually homogenized to disperse them evenly. Next, the evenly dispersed leeches are frozen again and mechanically homogenized. This allows for obtaining frozen and evenly dispersed leeches before freeze-drying, better preserving the antithrombin activity of the leeches and improving the product yield.

[0024] The method has the following advantages: (1) The frozen leech after the first freezing is in a partially frozen state with a soft exterior and a hard interior. It can be dispersed by manual rapid homogenization, avoiding the violent friction between hard materials, and will not introduce mechanical damage and disperse evenly. (2) Since the leech has been homogenized evenly before the second freezing, it promotes the uniformity of the frozen leech after the second freezing. It can be dispersed by rapid mechanical homogenization. At this time, the shear resistance is greatly reduced, which reduces the damage to the medicinal material caused by mechanical shear force (such as destroying the active structure of the leech tissue), and also improves the dispersion effect and shortens the homogenization time. (3) By controlling the process parameters such as the amount of water added, freezing temperature, hardness of the frozen material, homogenization speed and homogenization time in the two freezing processes, this invention can significantly improve the dispersion effect of the frozen leech and promote the subsequent freeze-drying effect. (4) In the study, this invention found that when the first freezing was controlled to a material hardness of 5000-6000g and the second freezing to a material hardness of 12000-14000g, the total homogenization time was only 4min. The resulting freeze-dried leech powder had high antithrombin activity. This may be because the dispersion effect of the frozen leech was better at the above hardness, and there was no loss of medicinal activity due to shear heating. In addition, the homogenization time was significantly shortened, which also avoided changes in the material properties and oxidation of the leech, thus comprehensively preserving the antithrombin activity of the leech. (5) The freeze-drying effect has a significant impact on the yield of freeze-dried leech powder. This invention promotes the subsequent freeze-drying effect by controlling various parameters in the pre-freezing and homogenization processes, thereby effectively improving the product yield.

[0025] 2. Compared to conventional methods that only remove the fishy smell from freeze-dried leech powder, this invention removes the fishy smell from both fresh leeches and freeze-dried leech powder in two steps. Fresh, unfrozen leeches have a strong fishy smell, so this invention removes the fishy smell from fresh leeches first, which can effectively improve the fishy smell removal effect.

[0026] In this invention, when removing the fishy smell from fresh leeches, an aqueous solution containing starch, deoxyarbutin, and fructose is used for soaking. Starch has a good adsorption effect on fishy substances and can effectively adsorb and remove fishy substances from the surface of the leeches. Deoxyarbutin and fructose not only improve the electrolyte level of the aqueous solution, which helps to release fishy substances from the leeches, but deoxyarbutin can also further bind to the fishy substances in the leeches and promote their removal. Fructose can also provide nutrients to the leeches and promote metabolism. The two combined promote the release and removal of fishy substances.

[0027] 3. In this invention, when deodorizing the leech powder obtained after freeze-drying and pulverizing, activated carbon is modified with alkali and organic acid to improve its adsorption effect. Then, volatile odor substances in the leech powder are blown into the activated carbon by a blower to promote adsorption. This method achieves non-contact adsorption between activated carbon and leech powder, avoiding the risk of residue and activity loss. It can also promote the removal of volatile odor substances in leech powder and better retain the active ingredients of leech powder.

[0028] The organic acid of the present invention is further preferably veratric acid, which can not only improve the adsorption effect of activated carbon, but also play a fixing role by combining with volatile fishy substances, preventing the leakage of volatile components adsorbed in activated carbon and affecting the deodorizing effect. Attached Figure Description

[0029] Figure 1 This is a flow chart illustrating the preparation process of the freeze-dried leech powder traditional Chinese medicine slices of the present invention; Figure 2 The image shows the product of the freeze-dried leech powder prepared according to this invention. Detailed Implementation

[0030] The above-mentioned solution will be further described below with reference to specific embodiments; it should be understood that these embodiments are used to illustrate the basic principles, main features and advantages of the present invention, and the present invention is not limited to the scope of the following embodiments; the implementation conditions used in the embodiments can be further adjusted according to specific requirements, and the implementation conditions not specified are usually the conditions in conventional experiments.

[0031] Unless otherwise specified in the following examples, all raw materials are commercially available or prepared by conventional methods in the art. Example 1

[0032] This embodiment provides a preparation process for freeze-dried leech powder, a traditional Chinese medicine decoction piece. The process flow is as follows: Figure 1 This includes the following steps: (1) Take fresh whole leeches, wash them, add water to the washed leeches at a mass percentage of 1.5%, mix them evenly, and place them at -8℃ for the first freezing. When the material hardness is 5500g, stop freezing and perform manual homogenization (stir repeatedly with a stainless steel grinding rod) for 3 minutes.

[0033] (2) Add water to the material obtained in step (1) at a mass percentage of 3.5%, mix evenly, and place it at -15℃ for a second freezing. When the material hardness is 13500g, stop freezing and perform mechanical homogenization at a speed of 6000r / min for 1min.

[0034] (3) The material obtained in step (2) is freeze-dried at a temperature of -45℃ for 25 hours and a vacuum of less than 20Pa. After freeze-drying, the material is pulverized, passed through a No. 3 (40-60 mesh) sieve, and sterilized to obtain freeze-dried leech powder. Example 2

[0035] The difference between this embodiment and embodiment 1 is that in step (1), the material is frozen to a hardness of 4000g. Example 3

[0036] The difference between this embodiment and embodiment 1 is that in step (1), the material is frozen to a hardness of 7000g. Example 4

[0037] The difference between this embodiment and embodiment 1 is that in step (2), the material is frozen to a hardness of 10000g. Example 5

[0038] The difference between this embodiment and embodiment 1 is that in step (2), the material is frozen to a hardness of 15000g. Example 6

[0039] Based on Example 1, this example further applies antioxidant and deodorizing treatment to the leeches to reduce the fishy smell of the freeze-dried leech powder.

[0040] This embodiment provides a preparation process for freeze-dried leech powder, a traditional Chinese medicine, including the following steps: (1) Take fresh whole leeches, wash them, add 4.5 times the weight of water and starch, mix them evenly, then add deoxyarbutin and fructose, soak for 35 minutes. The mass ratio of water, starch, deoxyarbutin and fructose is 1:0.08:0.01:0.03. After soaking, wash with clean water, then add water to the washed leeches at a mass percentage of 1.5%, mix evenly, and place at -8℃ for the first freezing. When the material hardness is 5500g, stop freezing and perform manual homogenization (stir repeatedly with a stainless steel grinding rod) for 3 minutes.

[0041] (2) Add water to the material obtained in step (1) at a mass percentage of 3.5%, mix evenly, and place it at -15℃ for a second freezing. When the material hardness is 13500g, stop freezing and perform mechanical homogenization at a speed of 6000r / min for 1min.

[0042] (3) The material obtained in step (2) is freeze-dried at a temperature of -45℃ for 25 hours and a vacuum of less than 20Pa. After freeze-drying, the material is pulverized and passed through a No. 3 (40-60 mesh) sieve to obtain Hirudo medicinalis powder.

[0043] (4) Soak activated carbon (100-180 mesh) in 5 times its mass of sodium hydroxide solution (mass concentration of 1.5%) for 1.5h, then take it out and dry it, soak it in 8 times its mass of veratril acid solution (containing 4.5% of veratril acid by mass) for 60min, then evaporate it to dryness to obtain modified activated carbon; take the leech powder obtained in step (3) and spread it on an 80-120 mesh sieve, then spread the modified activated carbon (mass of 50% of the leech powder) on a 200-300 mesh sieve and fix it directly above the sieve containing the leech powder, then blow air from below the sieve containing the leech powder at room temperature, with a wind speed of 7m / s and a blowing time of 30min.

[0044] (5) Sterilize the material obtained in step (4) to obtain freeze-dried leech powder, a traditional Chinese medicine. Example 7

[0045] This embodiment provides a preparation process for freeze-dried leech powder, a traditional Chinese medicine, including the following steps: (1) Take fresh whole leeches, wash them, add 3 times the weight of water and starch, mix them evenly, then add deoxyarbutin and fructose, soak for 20 minutes. The mass ratio of water, starch, deoxyarbutin and fructose is 1:0.05:0.005:0.02. After soaking, wash with clean water, then add 1% water to the washed leeches, mix evenly, and place at -10℃ for the first freezing. When the material hardness is 5000g, stop freezing and perform manual homogenization (stir repeatedly with a stainless steel grinding rod). The homogenization time is 4 minutes.

[0046] (2) Add water to the material obtained in step (1) at a mass percentage of 3%, mix evenly, and place at -20℃ for a second freezing. When the material hardness is 13000g, stop freezing and perform mechanical homogenization at a speed of 5000r / min for 2min.

[0047] (3) The material obtained in step (2) is freeze-dried at a temperature of -45℃ for 25 hours and a vacuum of less than 20Pa. After freeze-drying, the material is pulverized and passed through a No. 3 (40-60 mesh) sieve to obtain Hirudo medicinalis powder.

[0048] (4) Soak activated carbon (100-180 mesh) in 4 times its mass of sodium hydroxide solution (mass concentration of 1%) for 2 hours, then take it out and dry it, soak it in 5 times its mass of veratril acid solution (containing 3% of the mass of veratril acid) for 80 minutes, then evaporate it to dryness to obtain modified activated carbon; take the leech powder obtained in step (3) and spread it on an 80-120 mesh sieve, then spread the modified activated carbon (mass of 30% of the mass of leech powder) on a 200-300 mesh sieve and fix it directly above the sieve containing leech powder, then blow air from below the sieve containing leech powder at room temperature, with a wind speed of 5 m / s and a blowing time of 40 minutes.

[0049] (5) Sterilize the material obtained in step (4) to obtain freeze-dried leech powder, a traditional Chinese medicine. Example 8

[0050] This embodiment provides a preparation process for freeze-dried leech powder, a traditional Chinese medicine, including the following steps: (1) Take fresh whole leeches, wash them, add 6 times the weight of water and starch, mix them evenly, then add deoxyarbutin and fructose, soak for 50 minutes. The mass ratio of water, starch, deoxyarbutin and fructose is 1:0.1:0.02:0.04. After soaking, wash with clean water, then add 2% water to the washed leeches, mix evenly, and place at -5℃ for the first freezing. When the material hardness is 6000g, stop freezing and perform manual homogenization (stir repeatedly with a stainless steel grinding rod) for 5 minutes.

[0051] (2) Add water to the material obtained in step (1) at a mass percentage of 4%, mix evenly, and place at -10℃ for a second freezing. When the material hardness is 14000g, stop freezing and perform mechanical homogenization at a speed of 8000r / min for 3min.

[0052] (3) The material obtained in step (2) is freeze-dried at a temperature of -50°C for 24 hours and a vacuum of less than 20Pa. After freeze-drying, the material is pulverized and passed through a No. 3 (40-60 mesh) sieve to obtain Hirudo medicinalis powder.

[0053] (4) Soak activated carbon (100-180 mesh) in 6 times its mass of sodium hydroxide solution (mass concentration of 2%) for 1 hour, then take it out and dry it, soak it in 10 times its mass of veratril acid solution (containing 6% veratril acid by mass) for 40 minutes, then evaporate it to dryness to obtain modified activated carbon; take the leech powder obtained in step (3) and spread it on an 80-120 mesh screen, then spread the modified activated carbon (mass of 100% of the leech powder) on a 200-300 mesh screen and fix it directly above the screen containing the leech powder, then blow air from below the screen containing the leech powder at room temperature, with a wind speed of 10 m / s and a blowing time of 20 minutes.

[0054] (5) Sterilize the material obtained in step (4) to obtain freeze-dried leech powder, a traditional Chinese medicine. Example 9

[0055] The difference between this embodiment and embodiment 1 is that in step (4), the veratric acid solution is replaced with acetic acid solution. Example 10

[0056] The difference between this embodiment and embodiment 1 is that in step (4), the veratric acid solution is replaced with citric acid solution. Example 11

[0057] The difference between this embodiment and embodiment 1 is that in step (4), the veratric acid solution is replaced with vanillic acid solution.

[0058] I. Quality Inspection of the Freeze-dried Hirudo medicinal powder prepared by this invention The freeze-dried leech powder obtained by this invention is shown in [reference needed]. Figure 2 It can be seen that it is brownish-red, loose, and uniformly powdery.

[0059] Referring to the standard of Yunnan Province's Traditional Chinese Medicine Decoction Pieces (Yun YPBZ-0199-2013), the freeze-dried leech powder prepared in Examples 1-11 of this invention was subjected to quality testing. Its particle size, pH, microbial limit tests (Salmonella, bile-resistant Gram-negative bacteria, total aerobic bacteria, total mold and yeast count, Escherichia coli, etc.), heavy metal tests, and sulfur dioxide residue all met the standard requirements. Results for other components are shown in Table 1.

[0060] Table 1

[0061] As shown in Table 1, the freeze-dried leech powders prepared in Examples 1-11 of this invention all meet the relevant provisions of the "Yunnan Provincial Standard for Traditional Chinese Medicine Pieces" (Yun YPBZ-0199-2013), and the antithrombin activity is 1057.3-1285.8 U / g.

[0062] Among them, Examples 1-5 can effectively match the homogenization process by controlling the hardness of the frozen material during pre-freezing, and obtain freeze-dried leech powder with high antithrombin activity. In Example 1, the performance was better when the material hardness was controlled to 5500g and 13500g respectively, with an antithrombin activity as high as 1285.8 U / g.

[0063] II. Optimization of Pre-freezing and Homogenization Parameters in the Preparation Process of Freeze-dried Leech Powder To investigate the effects of the pre-freezing and homogenization processes of the present invention, as well as the effects of process parameters such as water addition, freezing temperature, hardness of the frozen material, homogenization speed, and homogenization time on the activity and yield of lyophilized Hirudo medicinalis powder, comparative examples 1-19 were set up respectively.

[0064] Comparative Example 1 The difference between this comparative example and Example 1 is that it uses a conventional pre-freezing and homogenization process.

[0065] The pre-freezing process is as follows: add 5% water to the cleaned leeches and freeze at -8℃ for 4 hours.

[0066] The homogenization process is as follows: the pre-frozen leeches are mechanically homogenized at a speed of 8000 r / min for 4 min.

[0067] Comparative Example 2 The difference between this comparative example and Example 1 is that in step (1), water is added to the washed leeches at a mass percentage of 0.5%.

[0068] Comparative Example 3 The difference between this comparative example and Example 1 is that in step (1), water is added to the washed leeches at a mass percentage of 3%.

[0069] Comparative Example 4 The difference between this comparative example and Example 1 is that in step (1), the freezing temperature is -2°C.

[0070] Comparative Example 5 The difference between this comparative example and Example 1 is that in step (1), the freezing temperature is -15°C.

[0071] Comparative Example 6 The difference between this comparative example and Example 1 is that in step (1), the material is frozen to a hardness of 3000g.

[0072] Comparative Example 7 The difference between this comparative example and Example 1 is that in step (1), the material is frozen to a hardness of 8000g.

[0073] Comparative Example 8 The difference between this comparative example and Example 1 is that in step (2), water is added to the material obtained in step (1) at a mass percentage of 2%.

[0074] Comparative Example 9 The difference between this comparative example and Example 1 is that in step (2), water is added to the material obtained in step (1) at a mass percentage of 5%.

[0075] Comparative Example 10 The difference between this comparative example and Example 1 is that in step (2), the freezing temperature is -6°C.

[0076] Comparative Example 11 The difference between this comparative example and Example 1 is that in step (2), the freezing temperature is -25°C.

[0077] Comparative Example 12 The difference between this comparative example and Example 1 is that in step (2), the material is frozen to a hardness of 9000g.

[0078] Comparative Example 13 The difference between this comparative example and Example 1 is that in step (2), the material is frozen to a hardness of 16000g.

[0079] Comparative Example 14 The difference between this comparative example and Example 1 is that in step (2), the rotation speed of the mechanical homogenizer is 2000 r / min.

[0080] Comparative Example 15 The difference between this comparative example and Example 1 is that in step (2), the rotation speed of the mechanical homogenizer is 15000 r / min.

[0081] Comparative Example 16 The difference between this comparative example and Example 1 is that in step (3), the freeze-drying temperature is -40°C.

[0082] Comparative Example 17 The difference between this comparative example and Example 1 is that in step (3), the freeze-drying temperature is -55°C.

[0083] Comparative Example 18 The difference between this comparative example and Example 1 is that in step (3), the freeze-drying time is 20 hours.

[0084] Comparative Example 19 The difference between this comparative example and Example 1 is that in step (3), the freeze-drying time is 28 hours.

[0085] Freeze-dried leech powder was prepared according to the method of Comparative Examples 1-19. The dispersion of the medicinal material during homogenization was observed, and the antithrombin activity and yield of the product were determined. The results were compared with those of Example 1, and the results are shown in Table 2. Among them, the yield (%) = 100% × (actual mass of freeze-dried leech powder / quantity of raw materials).

[0086] Table 2

[0087] As shown in Table 2, compared with Comparative Examples 1-15, in the preparation of lyophilized leech powder in Example 1 of the present invention, by optimizing the process parameters such as water addition, freezing temperature, hardness of frozen material, homogenization speed and homogenization time in the two freezing processes, the medicinal material is better dispersed during homogenization. Only a shorter homogenization time is needed to disperse the medicinal material evenly, reducing the loss of medicinal activity and promoting the subsequent freeze-drying effect, thereby effectively improving the product yield. As a result, the antithrombin activity and product yield of Example 1 are better than those of Comparative Examples 1-15.

[0088] Compared with Example 1, Comparative Examples 16-19 changed the freeze-drying temperature and time, resulting in a decrease in antithrombin activity and product yield.

[0089] III. Evaluation of the fishy smell of freeze-dried leech powder To investigate the effect of the deodorization method of the present invention on the sensory characteristics of the fishy smell of the finished freeze-dried leech powder, comparative examples 20-31 were set up respectively.

[0090] Comparative Example 20 The difference between this comparative example and Example 6 is that the whole fresh leech is not deodorized in step (1). That is, step (1) is: take the whole fresh leech, wash it, add water to the washed leech at a mass percentage of 1.5%, mix it evenly, and place it at -8℃ for the first freezing. When the material hardness is 5500g, stop freezing and perform manual homogenization for 3min.

[0091] Comparative Example 21 The difference between this comparative example and Example 6 is that the freeze-dried and pulverized leech powder is not deodorized, i.e., step (4) is not included.

[0092] Comparative Example 22 The difference between this comparative example and Example 6 is that step (4) directly uses modified activated carbon and leech powder to remove the fishy smell. Specifically, it includes: taking the leech powder obtained in step (3), adding 5 times the volume of water, then adding modified activated carbon (mass of 50% of the leech powder), shaking well, water bath (50°C) for 1 hour, drying at 45°C, separating the modified activated carbon and leech powder, and obtaining the product.

[0093] Comparative Example 23 The difference between this comparative example and Example 6 is that starch is not added in step (1).

[0094] Comparative Example 24 The difference between this comparative example and Example 6 is that deoxyarbutin is not added in step (1).

[0095] Comparative Example 25 The difference between this comparative example and Example 6 is that fructose is not added in step (1).

[0096] Comparative Example 26 The difference between this comparative example and Example 6 is that the activated carbon is not soaked in the veratric acid solution in step (4). The modified activated carbon is prepared by soaking the activated carbon in 5 times its mass of sodium hydroxide solution for 1.5 hours, and then taking it out and drying it.

[0097] Comparative Example 27 The difference between this comparative example and Example 6 is that in step (4), veratric acid is replaced with hydrochloric acid with a mass concentration of 1.5%.

[0098] Comparative Example 28 The difference between this comparative example and Example 6 is that in step (4), the wind speed of the blower is 3m / s.

[0099] Comparative Example 29 The difference between this comparative example and Example 6 is that in step (4), the wind speed of the blower is 15 m / s.

[0100] Comparative Example 30 The difference between this comparative example and Example 6 is that in step (4), the blowing time is 10 minutes.

[0101] Comparative Example 31 The difference between this comparative example and Example 6 is that in step (4), the blowing time is 50 min.

[0102] The fishy smell of leeches was removed using the methods of Comparative Examples 20-31, and freeze-dried leech powder was prepared. The fishy smell was evaluated. Then, the antithrombin activity of the product was measured, the antithrombin activity retention rate of the deodorization method was calculated, and the results were compared with those of Example 6. The results are shown in Table 3.

[0103] The method for evaluating the fishy smell was as follows: Ten evaluators (aged 20-25, half male and half female) were randomly selected to form an evaluation group to conduct sensory evaluation of the freeze-dried leech powder and calculate the average score. The fishy smell evaluation used a 10-point scale, with odorless distilled water scoring 0 points and untreated freeze-dried leech powder scoring 10 points. During the evaluation, 1 g of freeze-dried leech powder sample was placed in a sample cup, warm water (50±2℃) was added, and the mixture was stirred evenly. The evaluators conducted sensory evaluation according to the above scoring rules, and the average score was taken as the final score. Score 0-2: virtually no fishy smell; Score 2-4: slightly fishy smell; Score 4-6: fishy smell; Score 6-8: obvious fishy smell; Score 8-10: strong fishy smell.

[0104] Antithrombin activity retention rate (%) = 100% × (antithrombin activity of the product after deodorization / antithrombin activity of the product before deodorization, i.e., Example 1).

[0105] Table 3

[0106] As shown in Table 3, compared with Comparative Examples 20-31, Example 6 of the present invention uses a specific method to remove the fishy smell from fresh leeches and freeze-dried leech powder in two steps, which can effectively remove the fishy smell of the product, and the antithrombin activity retention rate is 98.05%, with no significant loss of activity.

[0107] In Comparative Example 22, modified activated carbon was mixed with leech powder to remove the fishy smell. Due to the strong adsorption capacity of the modified activated carbon, it adsorbed the active ingredients of the product along with the fishy smell, resulting in a significant decrease in the retention rate of antithrombin activity. Comparative Examples 29 and 31 increased the blowing speed and blowing time, respectively, and the antithrombin activity also showed a certain loss.

[0108] Finally, it should be noted that the above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A preparation process of a freeze-dried powder of traditional Chinese medicine decoction pieces of Lachesis muta, characterized in that: Includes the following steps: Take fresh whole leeches, wash them, pre-freeze them, homogenize them, freeze-dry them, pulverize them, sieve them, and sterilize them to obtain the product; Pre-freezing and homogenization include: (1) Add water to the cleaned leeches, mix well and place at -5-10℃ for the first freezing. When the material hardness is 4000-7000g, stop freezing and perform manual homogenization for 3-5 minutes. (2) Add water to the material obtained in step (1), mix evenly, and place it at -(10-20)℃ for a second freezing. When the material hardness is 10000-15000g, stop freezing and perform mechanical homogenization at a speed of 3000-10000r / min for 1-3min. The amount of water added in step (1) is 1-2% of the mass of the washed leeches, and the amount of water added in step (2) is 3-4% of the mass of the material obtained in step (1).

2. The manufacturing process of claim 1, wherein: In step (1), the material is frozen until the hardness is 5000-6000g, and the homogenization time is 3min.

3. The manufacturing process of claim 1, wherein: In step (2), the material is frozen until the hardness is 13000-14000g, the homogenization speed is 5000-8000r / min, and the homogenization time is 1min.

4. The manufacturing process of claim 1, wherein: The freeze-drying temperature is -45-50℃, the time is 23-26h, and the sieve used is No. 3 sieve.

5. A method for removing fishy smell in the preparation process of the freeze-dried traditional Chinese medicine decoction pieces of Hippopotamus amphibius Linnaeus according to claim 1, characterized in that: The method for removing fishy odors includes a first deodorization treatment and a second deodorization treatment; The first deodorization treatment is to remove the fishy smell from fresh leeches. The specific steps include: after washing the leeches, add water and starch, mix well, then add deoxyarbutin and fructose, soak for 20-50 minutes, and then wash. The second deodorization process involves removing the odor from the freeze-dried and pulverized leech powder. Specific steps include: soaking activated carbon in a sodium hydroxide solution, then removing and drying it; soaking it in an organic acid solution, then evaporating it to dryness to obtain modified activated carbon; spreading the freeze-dried and pulverized leech powder evenly on a sieve; then spreading the modified activated carbon evenly on the sieve and fixing it directly above the sieve containing the leech powder; and finally, blowing air from below the sieve containing the leech powder. In the second deodorization treatment, the organic acid is veratric acid, and the blowing conditions include: wind speed of 5-10 m / s and blowing time of 20-40 min.

6. The method of claim 5, wherein: In the first deodorization process, the mass ratio of water, starch, deoxyarbutin, and fructose is 1:(0.05-0.1):(0.005-0.02):(0.02-0.04), and the amount of water added is 3-6 times the mass of the washed leeches.

7. The method of claim 5, wherein: In the second deodorization treatment, the activated carbon is 100-180 mesh, the mass concentration of sodium hydroxide solution is 1-2%, the soaking time of sodium hydroxide solution is 1-2 hours, and the soaking time of organic acid solution is 40-80 minutes. The powdered Hirudo nipponia powder was spread on a sieve of 80-120 mesh, and the modified activated carbon was spread on a sieve of 200-300 mesh. The mass of the modified activated carbon on the sieve was 10-100% of the mass of the Hirudo nipponia powder.

8. The method of claim 5, wherein: In the second deodorization process, the mass percentage of the organic acid in the organic acid solution is 3-6%, the solvent in the organic acid solution is one or several of water, ethanol, diethyl ether, and acetone, and the amount of the organic acid solution added is 5-10 times the mass of the activated carbon after being soaked in the sodium hydroxide solution and dried.

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