Composite extraction process of silkworm moth and herba epimedii and application of composite extraction process in preparation of silkworm, antler and Tian capsules

Abstract

The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to a compound extraction process of silkworm moth and herba epimedii and application of the compound extraction process in preparation of silkworm, antler and Tian capsules. The extraction process comprises the following steps: (1) weighing the following raw materials according to a formula; (2) mixing male silkworm moth, pilose antler, penis et testis cervi and centipede to obtain animal medicinal materials, performing water extraction, and separating to obtain filtrate and medicine residues; mixing the remaining raw materials to obtain a plant medicinal material, performing CO2 supercritical extraction, and collecting volatile oil and extraction residues; (3) mixing the medicine residues and the extraction residues in the step (2), and performing anaerobic fermentation to obtain an extracting solution; and (4) mixing the filtrate in the step (2) and the extracting solution in the step (3), concentrating under reduced pressure to obtain clear paste, adding volatile oil, and uniformly stirring to obtain a composite extraction product. According to the compound extraction process, the synergistic effect of active substances in the product is improved by optimizing the extraction steps, and the medicine effect is improved when the compound extraction process is used for preparing the silkworm, antler and Zhuanday capsules.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of traditional Chinese medicine, and particularly relates to a compound extraction process of silkworm moth and herba epimedii and application thereof in preparation of silkworm chitinous capsule. BACKGROUND

[0002] Silkworm chitinous capsule is a kind of traditional Chinese medicine. The components of the medicine are quite rich. The formula is derived from Jing Yue Quan Shu by Zhang Jingyue, and combines two classic famous prescriptions, Yougui Pill and Zanyu Dan, including 17 kinds of Chinese medicinal materials: silkworm moth (processed), herba epimedii (moxibustion with sheep oil), leafflower root, prepared rehmannia root, Chinese yam, dogwood fruit, medlar fruit, dodder seed (steamed), deer horn, eucommia bark (charred), angelica root (fried), cassia bark, monkshood root (processed), centipede (alcohol-processed), gastrodia elata, ginseng, and deer penis. Modern pharmacological studies have shown that the product has certain androgenic activity. It has the effects of warming and tonifying kidney yang and filling essence and blood, and is used for patients with long-term illness, fatigue, poor appetite, incontinence, aversion to cold, cold limbs, and weak knees. It is often used in the treatment of impotence caused by deficiency of kidney yang and essence, and deficiency of the penis. Symptoms include impotence, erectile dysfunction, pale complexion, dizziness, fatigue, weakness, and soft knees. The tongue is pale and the pulse is thin.

[0003] In modern society, although silkworm chitinous capsule has shown significant effects in tonifying yang and relieving essence deficiency, its effect in tonifying kidney and preserving essence is not satisfactory in the face of environmental pollution, food safety problems, and life pressure. Modern research shows that environmental pollution such as harmful substances in the air, heavy metal pollution in water sources, and pesticide residues in the soil can negatively affect male reproductive health, leading to a decrease in sperm count and quality. At the same time, food safety problems, including the misuse of pesticides, hormones, and antibiotics, have also exacerbated this trend. These factors not only affect the overall health status of the human body, but also can interfere with the delicate hormonal balance in the body, thereby weakening the expected effect of traditional Chinese medicine formulas in tonifying the kidney and preserving essence. In addition, the sustained high-pressure environment brought about by modern lifestyle cannot be ignored. Long-term work pressure, irregular life and rest, and lack of enough rest time make many people long-term in sub-health state, which is particularly detrimental to reproductive health. In addition, the generally insufficient amount of exercise leads to poor blood circulation, especially in the pelvic region, which further affects the normal interaction between the kidney and the reproductive system.

[0004] With the general decline in the number and quality of male sperm, there is an increasing demand for traditional Chinese medicine that can effectively improve this situation. However, due to the above reasons, the traditional silkworm chitinous capsule prepared by water extraction faces new challenges in adapting to the characteristics of modern population, and the therapeutic effect cannot meet the requirements of patients. SUMMARY

[0005] The application aims to provide a composite extraction process of silkworm moth and epimedium and application in preparation of Zhongtian capsule.

[0006] To achieve the above-mentioned purpose, the application provides the following technical scheme. A composite extraction process of silkworm moth and epimedium comprises the following steps. (1) The following raw materials are weighed according to the formula: epimedium, radix morindae officinalis, radix rehmanniae preparata, silkworm moth, semen cuscutae, cornus officinalis, eucommia ulmoides, angelica sinensis, medlar, cassia bark, ginseng, gastrodia elata, pilose antler, aconite, centipede, Chinese yam, deer penis; (2) The male silkworm moth, pilose antler, deer penis and centipede are mixed to obtain animal medicinal materials, which are subjected to water extraction to obtain filtrate and dregs; the remaining raw materials are mixed to obtain plant medicinal materials, which are subjected to CO2 supercritical extraction to collect volatile oil and extraction residues; (3) The dregs and extraction residues of step (2) are mixed to obtain fermentation raw materials, which are mixed with culture medium and strains, subjected to anaerobic fermentation, sterilized, filtered and then an extraction liquid is obtained; (4) The filtrate of step (2) and the extraction liquid of step (3) are mixed, concentrated under reduced pressure to obtain a clear paste, the volatile oil is added and stirred uniformly to obtain a composite extraction product.

[0007] Preferably, the following raw materials are weighed according to the formula: epimedium 10-20 parts, radix morindae officinalis 7-13 parts, radix rehmanniae preparata 8-15 parts, silkworm moth 3-9 parts, semen cuscutae 8-15 parts, cornus officinalis 9-17 parts, eucommia ulmoides 3-9 parts, angelica sinensis 6-12 parts, medlar 7-14 parts, cassia bark 2-9 parts, ginseng 1-10 parts, gastrodia elata 3-11 parts, pilose antler 1-10 parts, aconite 4-10 parts, centipede 1-6 parts, Chinese yam 10-17 parts, deer penis 1-8 parts.

[0008] Preferably, the male silkworm moth, pilose antler, deer penis and centipede are mixed to obtain animal medicinal materials, which are crushed, soaked in water, heated to 95-100 DEG C, kept for 1-2 h, and repeatedly extracted for 2-4 times, and the filtrate is collected and the dregs are obtained.

[0009] Preferably, the remaining raw materials are mixed to obtain plant medicinal materials, which are crushed to less than 200 meshes, subjected to CO2 supercritical extraction, the extraction pressure is 30-35 MPa, the extraction temperature is 40-50 DEG C, the extraction time is 1-3 h, and the volatile oil and extraction residues are collected.

[0010] The preparation method of Zhongtian capsule in the prior art is water extraction of all raw materials, and the treatment effect is not ideal for diseases with little sperm or low sperm motility. Even if the dregs are further extracted to obtain more active ingredients, the improvement effect is not good. The composition of the raw materials is optimized in the application, and the animal raw materials and plant raw materials are extracted respectively, so that the contents of FSH, LH and T in the serum can be improved.

[0011] Preferably, the amount of the bacterial strain is 5.0*10 6- 9.0*10 7 CFU / L of the culture medium.

[0012] Preferably, the anaerobic fermentation conditions are 35-38 DEG C anaerobic fermentation for 3-5 days.

[0013] Preferably, the bacterial strain comprises Lactobacillus bifidus, Lactobacillus brevis and Lactobacillus rhamnosus in a ratio of 1: (1.2-1.5): (0.6-0.8) in terms of the number of viable bacteria.

[0014] The product obtained by fermenting the extracted residues with specific bacterial strains can improve the activity of sperm.

[0015] Preferably, the culture medium formula is: 12-15 g of proteose peptone, 35-40 g of glucose, 1.5-2.0 g of ammonium citrate, 1000 mL of distilled water, and the pH is adjusted to neutral.

[0016] Preferably, the mass ratio of Epimedium, Cornu Cervi and Panax ginseng is (12-17):(2-6):(3-7).

[0017] Preferably, the following raw materials are weighed according to the formula: Epimedium 15 parts, Radix Morindae Officinalis 10 parts, Radix Rehmanniae Preparata 12 parts, moth 8 parts, Semen Euryale 10 parts, Fructus Corni 10 parts, Eucommia ulmoides 8 parts, Angelica 8 parts, Fructus Schisandrae Chinensis 10 parts, Cinnamomum cassia 6 parts, Panax ginseng 5 parts, Gastrodia 6 parts, Cornu Cervi 4 parts, Aconiti Lateralis Radix 6 parts, Solpugid 3 parts, Dioscorea 12 parts, Deer penis 4 parts.

[0018] The application provides application of the compound extraction process of the moth and Epimedium in preparation of a capsule of the silkworm chrysalis and Epimedium.

[0019] The above raw materials are strictly processed according to the Chinese Pharmacopoeia (2020 edition) and traditional processing specifications.

[0020] Compared with the prior art, the application has the following advantages and beneficial effects: 1. On the basis of the traditional classic prescription, the raw material formula of the silkworm chrysalis and Epimedium capsule is optimized according to the physical characteristics and health needs of modern people, the ratio of various medicinal materials is scientifically adjusted, the synergistic effect between the components is enhanced, the medicine can not only warm and invigorate the kidney yang and fill the essence and blood, but also more effectively play the roles of tonifying the kidney and consolidating essence, improving the number and quality of sperm, etc. The new formula not only retains the significant curative effects of the original prescription in improving impotence, fatigue, soreness of the waist and knees, etc., but also further enhances the intervention ability on the common problems of modern people, such as deficiency of kidney essence and reduction of reproductive function, thereby improving the overall efficacy and clinical application value.

[0021] 2. In the preparation process, the application enhances the synergistic effect between different effective components by innovating the process, so that the final prepared caterpillar hair and Gouteng capsule has higher bioavailability and stronger pharmacological effect. The improvement of the process effectively solves the problems of large loss of active components, incomplete extraction, poor component synergy and the like in the traditional extraction method. DETAILED DESCRIPTION

[0022] The technical solutions in the embodiments of the application will be described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the application, rather than all the embodiments. Based on the embodiments in the application, all other embodiments obtained by those skilled in the art without creative labor fall within the protection scope of the application.

[0023] The raw materials used in the following embodiments of the application are all commercially available goods: Lactobacillus coryniformis, item number HZB231387, Zhengzhou Fangjue Biotechnology Co., Ltd. Lactobacillus plantarum, item number BNCC361473, Beinaeliang Biotechnology Co., Ltd. Lactobacillus delbrueckii, item number HZB139506, Wuhan Huizao Biotechnology Co., Ltd. Lactobacillus brevis, strain number HZB134658, Zhengzhou Fangjue Biotechnology Co., Ltd. Lactobacillus rhamnosus, strain number HZB196554, Zhengzhou Fangjue Biotechnology Co., Ltd. Lactobacillus bulgaricus, item number HZB116345, Zhengzhou Fangjue Biotechnology Co., Ltd.

[0024] Embodiment 1 The embodiment provides a compound extraction process of silkworm moth and Epimedium, which comprises the following steps: (1) The following raw materials are weighed according to the formula: Epimedium 15 parts, Radix Morindae Officinalis 10 parts, Radix Rehmanniae Preparata 12 parts, male silkworm moth 8 parts, Semen Trichosanthis 10 parts, Fructus Corni 10 parts, Eucommia ulmoides Oliv. 8 parts, Angelica sinensis 8 parts, Fructus Lycii 10 parts, Cinnamomum cassia 6 parts, Panax ginseng 5 parts, Gastrodia elata 6 parts, Cornu Cervi Pantotrichum 4 parts, Aconitum carmichaeli 6 parts, Solenopsis geminata 3 parts, Dioscorea opposita 12 parts, Cornu Cervi 4 parts; (2) The male silkworm moth, Cornu Cervi Pantotrichum, Cornu Cervi and Solenopsis geminata are mixed to obtain animal medicinal materials, which are crushed to less than 40 meshes, 8 times the amount of purified water is added, soaked at 60℃ for 40 min, heated to reflux extraction for 1.5 h, repeated extraction for 3 times, and the filtrate is filtered and combined to obtain the residue; The remaining raw materials are mixed to obtain plant medicinal materials, which are crushed to less than 200 meshes, and subjected to CO2 supercritical extraction, with an extraction pressure of 32 MPa, an extraction temperature of 45℃, and an extraction time of 2 h, and the volatile oil and extraction residue are collected; (3) mixing the residues of step (2) and the extraction residues to obtain fermentation raw materials, mixing the fermentation raw materials, culture medium and bacterial strains, the adding ratio of the fermentation raw materials and the culture medium being 17 g:1 L, and the amount of the bacterial strains being 4.0 x 10 7 CFU / L of culture medium, the bacterial strains including Lactobacillus sanfilippo, Lactobacillus brevis and Lactobacillus rhamnosus in a ratio of 1:1.4:0.7 in terms of the number of viable bacteria; anaerobic fermentation at 37°C for 4 days; the culture medium formula being: peptone 12 g, glucose 35 g, ammonium citrate 1.5 g, distilled water 1000 mL, adjusting the pH to neutral, and high-pressure sterilization at 121°C for 20 min. After the fermentation, sterilization and filtration are performed to obtain an extract; (4) mixing the filtrate of step (2) and the extract of step (3), concentrating under reduced pressure to a clear paste with a relative density of 1.15 at 60°C, adding volatile oil, and stirring uniformly to obtain a composite extraction product.

[0025] Example 2 The difference between this example and Example 1 is that the bacterial strains include Lactobacillus sanfilippo, Lactobacillus brevis and Lactobacillus rhamnosus in a ratio of 1:1.2:0.8 in terms of the number of viable bacteria.

[0026] Example 3 The difference between this example and Example 1 is that the bacterial strains include Lactobacillus sanfilippo, Lactobacillus brevis and Lactobacillus rhamnosus in a ratio of 1:1.5:0.6 in terms of the number of viable bacteria.

[0027] Example 4 The difference between this example and Example 1 is that the following raw materials are weighed according to the formula: Herba Epimedii 12 parts, Radix Morindae Officinalis 13 parts, Radix Rehmanniae Preparata 8 parts, Bombyx Batryticatus 9 parts, Semen Nelumbinis 15 parts, Fructus Corni 17 parts, Eucommia Ulmoides 3 parts, Angelica sinensis 12 parts, Fructus Lycii 7 parts, Cinnamomum Cassia 2 parts, Radix Ginseng 7 parts, Gastrodia Elata 11 parts, Cornu Cervi Pantotrichum 2 parts, Aconiti Lateralis Radix 4 parts, Solenopsis geminata 6 parts, Dioscorea Opposita 10 parts, Cornu Cervi 8 parts.

[0028] Comparative Example 1 The difference between this comparative example and Example 1 is that the following raw materials are weighed according to the formula: Herba Epimedii 5 parts, Radix Morindae Officinalis 15 parts, Radix Rehmanniae Preparata 3 parts, Bombyx Batryticatus 14 parts, Semen Nelumbinis 18 parts, Fructus Corni 6 parts, Eucommia Ulmoides 12 parts, Angelica sinensis 4 parts, Fructus Lycii 3 parts, Cinnamomum Cassia 11 parts, Radix Ginseng 15 parts, Gastrodia Elata 2 parts, Cornu Cervi Pantotrichum 15 parts, Aconiti Lateralis Radix 1 part, Solenopsis geminata 7 parts, Dioscorea Opposita 4 parts, Cornu Cervi 10 parts.

[0029] Comparative Example 2 The difference between the present comparative example and example 1 is that the following raw materials are weighed according to the formula: Herba Epimedii 25 parts, Radix Morindae Officinalis 2 parts, Radix Rehmanniae Preparata 17 parts, Bombyx Batryticatus 1 part, Semen Leonuri 5 parts, Fructus Corni 20 parts, Eucommia Ulmoides 1 part, Angelica 15 parts, Fructus Lycii 17 parts, Cinnamomum Cassia 1 part, Ginseng 2 parts, Gastrodia 1 part, Cornu Cervi Pantotrichum 1 part, Aconiti Praeparata 12 parts, Solpugid 1 part, Dioscorea 20 parts, Cornu Cervi 1 part.

[0030] Comparative Example 3 The difference between the present comparative example and example 1 is that the total parts of Herba Epimedii, Cornu Cervi Pantotrichum and Ginseng are unchanged, but the ratio is changed.

[0031] The following raw materials are weighed according to the formula: Herba Epimedii 10 parts, Radix Morindae Officinalis 10 parts, Radix Rehmanniae Preparata 12 parts, Bombyx Batryticatus 8 parts, Semen Leonuri 10 parts, Fructus Corni 10 parts, Eucommia Ulmoides 8 parts, Angelica 8 parts, Fructus Lycii 10 parts, Cinnamomum Cassia 6 parts, Ginseng 7 parts, Gastrodia 6 parts, Cornu Cervi Pantotrichum 7 parts, Aconiti Praeparata 6 parts, Solpugid 3 parts, Dioscorea 12 parts, Cornu Cervi 4 parts.

[0032] Comparative Example 4 The difference between the present comparative example and example 1 is that the total parts of Herba Epimedii, Cornu Cervi Pantotrichum and Ginseng are unchanged, but the ratio is changed.

[0033] The following raw materials are weighed according to the formula: Herba Epimedii (fried with mutton tallow) 22 parts, Radix Morindae Officinalis 10 parts, Radix Rehmanniae Preparata 12 parts, Bombyx Batryticatus (processed) 8 parts, Semen Leonuri (steamed) 10 parts, Fructus Corni 10 parts, Eucommia Ulmoides (charred) 8 parts, Angelica (fried) 8 parts, Fructus Lycii 10 parts, Cinnamomum Cassia 6 parts, Ginseng 1 part, Gastrodia 6 parts, Cornu Cervi Pantotrichum 1 part, Aconiti Praeparata (processed) 6 parts, Solpugid (processed with wine) 3 parts, Dioscorea 12 parts, Cornu Cervi 4 parts.

[0034] Comparative Example 5 The difference between the present comparative example and example 1 is that the strains include Lactobacillus coryniformis, Lactobacillus brevis and Lactobacillus rhamnosus in a ratio of 1:1.1:0.9.

[0035] Comparative Example 6 The difference between the present comparative example and example 1 is that the strains include Lactobacillus coryniformis, Lactobacillus brevis and Lactobacillus rhamnosus in a ratio of 1:1.6:0.5.

[0036] Comparative Example 7 The difference between the present comparative example and example 1 is that the strains include Lactobacillus plantarum, Lactobacillus delbrueckii and Lactobacillus bulgaricus in a ratio of 1:1.4:0.7.

[0037] Comparative Example 8 The difference between the present comparative example and example 1 is that all the raw materials are mixed for extraction.

[0038] A composite extraction process of Bombyx Batryticatus and Herba Epimedii, comprising the following steps: (1) Take the following mass parts of raw materials according to the formula: Herba Epimedii 15 parts, Radix Morindae Officinalis 10 parts, Radix Rehmanniae Preparata 12 parts, Male silkworm moth 8 parts, Semen Trichosanthis 10 parts, Fructus Corni 10 parts, Eucommia ulmoides 8 parts, Angelica 8 parts, Fructus Lycii 10 parts, Cinnamon 6 parts, Ginseng 5 parts, Gastrodia 6 parts, Cornu Cervi Pantotrichum 4 parts, Aconiti Lateralis Radix 6 parts, Solpugid 3 parts, Dioscorea 12 parts, Deer penis 4 parts; (2) Mix all the raw materials, crush to less than 40 meshes, add 8 times the amount of purified water, soak at 60°C for 40 min, heat reflux extraction for 1.5 h, repeat extraction 3 times, filter and combine the filtrate, and at the same time obtain the dregs; (3) The dregs of step (2) are used as fermentation raw materials, and the fermentation raw materials, culture medium and strains are mixed, the addition ratio of fermentation raw materials and culture medium is 17g:1L, and the amount of strains is 4.0×10 7 CFU / L of culture medium, and the strains include Lactobacillus coryniformis, Lactobacillus brevis and Lactobacillus rhamnosus with a ratio of 1:1.4:0.7 in the number of viable bacteria; anaerobic fermentation at 37°C for 4 days; the culture medium formula is: peptone 12g, glucose 35g, ammonium citrate 1.5g, distilled water 1000mL, adjust pH to neutral with 15wt% sodium hydroxide solution, autoclave at 121°C for 20min. After fermentation, sterilize and filter to obtain the extract; (4) Mix the filtrate of step (2) and the extract of step (3), concentrate under reduced pressure to a clear paste with a relative density of 1.15 at 60°C, stir evenly to obtain the composite extraction product.

[0039] Comparative Example 9 The difference between this comparative example and Example 1 is that the plant raw materials are extracted by water extraction method.

[0040] A composite extraction process of silkworm moth and Herba Epimedii, comprising the following steps: (1) Take the following mass parts of raw materials according to the formula: Herba Epimedii 15 parts, Radix Morindae Officinalis 10 parts, Radix Rehmanniae Preparata 12 parts, Male silkworm moth 8 parts, Semen Trichosanthis 10 parts, Fructus Corni 10 parts, Eucommia ulmoides 8 parts, Angelica 8 parts, Fructus Lycii 10 parts, Cinnamon 6 parts, Ginseng 5 parts, Gastrodia 6 parts, Cornu Cervi Pantotrichum 4 parts, Aconiti Lateralis Radix 6 parts, Solpugid 3 parts, Dioscorea 12 parts, Deer penis 4 parts; (2) Mix male silkworm moth, Cornu Cervi Pantotrichum, deer penis and Solpugid to obtain animal medicinal materials, crush to less than 40 meshes, add 8 times the amount of purified water, soak at 60°C for 40 min, heat reflux extraction for 1.5 h, repeat extraction 3 times, filter and combine the filtrate to obtain filtrate 1, and at the same time obtain dregs 1; Mix the remaining raw materials to obtain plant medicinal materials, crush to less than 200 meshes, add 8 times the amount of purified water, soak at 60°C for 40 min, heat reflux extraction for 1.5 h, repeat extraction 3 times, filter and combine the filtrate to obtain filtrate 2, and at the same time obtain dregs 2; (3) mixing the residues 1 and residues 2 of step (2) to obtain fermentation raw materials, mixing the fermentation raw materials, culture medium and strains, the adding ratio of the fermentation raw materials and the culture medium is 17g:1L, the amount of the strains is 4.0x10 7 CFU / L of culture medium, the strains include Lactobacillus mali, Lactobacillus brevis and Lactobacillus rhamnosus in a ratio of 1:1.4:0.7 in terms of the number of viable bacteria; anaerobic fermentation at 37℃ for 4 days; the culture medium formula is: peptone 12g, glucose 35g, ammonium citrate 1.5g, distilled water 1000mL, adjust pH to neutral with 15wt% sodium hydroxide solution, high pressure sterilization at 121℃ for 20min. After fermentation, sterilization and filtration, an extract is obtained; (4) mixing the filtrate 1 and filtrate 2 of step (2) and the extract of step (3) to obtain a clear extract with a relative density of 1.15 at 60℃ under reduced pressure, stirring uniformly to obtain a composite extraction product.

[0041] Comparative Example 10 The difference between this comparative example and Example 1 is that the fermentation raw materials are subjected to alcohol extraction.

[0042] A composite extraction process of silkworm moth and epimedium, comprising the following steps: (1) taking the following raw materials according to the formula: epimedium 15 parts, radix morindae officinalis 10 parts, radix rehmanniae preparata 12 parts, male silkworm moth 8 parts, semen cuscutae 10 parts, fructus corni 10 parts, eucommia ulmoides oliver 8 parts, angelica sinensis 8 parts, medlar 10 parts, cinnamomum cassia 6 parts, panax ginseng 5 parts, gastrodia elata 6 parts, pilose antler 4 parts, aconite 6 parts, centipede 3 parts, Chinese yam 12 parts, deer penis 4 parts; (2) mixing the male silkworm moth, pilose antler, deer penis and centipede to obtain animal medicinal materials, crushing to less than 40 mesh, adding 8 times the amount of purified water, soaking at 60℃ for 40min, heating to reflux extraction for 1.5h, repeating extraction for 3 times, filtering and combining the filtrate to obtain residues; mixing the remaining raw materials to obtain plant medicinal materials, crushing to less than 200 mesh, performing CO2 supercritical extraction, extraction pressure 32MPa, extraction temperature 45℃, extraction time 2h, collecting volatile oil and extraction residues; (3) mixing the residues of step (2) and the extraction residues to obtain fermentation raw materials, adding 8 times the volume fraction of 80% ethanol aqueous solution, reflux extraction for 1.5h, repeating extraction for 3 times, filtering and combining the filtrate to obtain an extract; (4) mixing the filtrate of step (2) and the extract of step (3), concentrating to a clear extract with a relative density of 1.15 at 60℃ under reduced pressure, adding volatile oil, stirring uniformly to obtain a composite extraction product.

[0043] Comparative Example 11 The comparative example is CN 110051784 A, a new application of a traditional Chinese medicine composition in the preparation of a medicine for treating and preventing kidney yang deficiency syndrome.

[0044] Performance test SD rats, male, SPF level, body weight 180-220g, from Shanghai Slake Experimental Animal Co., Ltd.; After adaptive feeding for 7 days, group into groups, 8 rats per group.

[0045] The grouping is as follows: Blank control group: no treatment.

[0046] Model group: give gossypol acetic acid 50mg / kg, once a day, continuous administration for 3 days. Start giving saline 1.2mL / kg on the 4th day, once a day for 3 weeks.

[0047] Experimental group: give gossypol acetic acid 50mg / kg, once a day, continuous administration for 3 days. Start giving the products of examples 1-4 and comparative examples 1-11 0.2g / kg on the 4th day, 1.2mL / Kg after dilution with saline, once a day for 3 weeks.

[0048] Positive control group: first give gossypol acetic acid 50mg / kg, once a day, continuous administration for 3 days. Start giving methyltestosterone 25mg / kg on the 4th day, 1.2mL / Kg after dilution with saline, once a day for 3 weeks.

[0049] (1) After the last administration, the rats were fasted for 16h, anesthetized with pentobarbital (45mg / kg), and blood was collected from the abdominal aorta, centrifuged, and serum was separated. The contents of follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone (T) were detected by radioimmunoassay.

[0050] (2) Put a test tube containing 6mL of saline in a 37℃ water bath, then cut the right testis and put it into the test tube, shake well, and incubate at 37℃ for 15min to fully dissociate the sperm. Take a drop and count it with a hemocytometer. Take another drop and smear it, observe the sperm motility, and observe 300 sperm, count the number of motile and non-motile sperm respectively.

[0051] Sperm motility = number of motile sperm / total number of sperm x 100%.

[0052] The results are shown in Table 1.

[0053] Table 1 Test results

[0054] From Table 1, it can be seen that the product prepared by the composite extraction process of Examples 1-4 can increase the contents of FSH, LH and T, help to promote sperm production, stimulate secretory hormones, maintain male characteristics, and also increase sperm activity.

[0055] In Comparative Examples 1 and 2, the composition of the raw materials is changed, and the spermatogenic and sperm activity of the product decrease, indicating that the synergistic effect of the raw materials of the formula of the present application is higher.

[0056] In Comparative Examples 3 and 4, the ratio of Epimedium, Cornu Cervi and Ginseng in the raw materials is changed, and the sperm activity decreases significantly. The inventors found that the ratio of these three traditional Chinese medicines in the formula has an important influence on sperm activity. Analysis shows that Epimedium can enhance the activity of Leydig cells, increase the energy supply for spermatogenesis, Cornu Cervi can promote the proliferation of spermatogenic cells, and Ginseng can inhibit the apoptosis of spermatogenic cells. Under the specific ratio in the formula of the present application, the three can better play a synergistic effect, and the change of the ratio breaks the balance of the synergistic effect, resulting in a decrease in sperm activity.

[0057] In Comparative Examples 5-7, the type and ratio of the strains are changed, which can lead to a decrease in the contents of FSH, LH and T, indicating that the active ingredients are contained in the residue after extraction, and only the use of specific strains for fermentation can make all components in the product have the best synergistic effect. In Comparative Examples 8-9, the extraction method is changed, which can lead to a decrease in the contents of FSH, LH and T, indicating that the scheme of the present application is not only to increase the extraction amount of active components, but more importantly, through a reasonable extraction process, the synergism of effective components in the product is achieved.

[0058] The above describes the preferred embodiments of the present application. It should be noted that for those skilled in the art, without departing from the principles of the present application, several improvements and refinements can be made, and these improvements and refinements should also be considered as the protection scope of the present application.

Claims

1. A combined extraction process of silkworm moth and Epimedium, characterized in that, It comprises the following steps: (1) the following ingredients are weighed according to the formula: Herba Epimedii, Radix Morindae Officinalis, Radix Rehmanniae Preparata, silkworm moth, Semen Leonuri, Fructus Corni, Eucommia ulmoides, Angelica sinensis, Fructus Lycii, Cinnamomum cassia, ginseng, Gastrodia elata, Cornu Cervi Pantotrichum, Aconitum, centipede, Dioscorea opposita, Cornu Cervi; (2) the silkworm moth, Cornu Cervi, Cornu Cervi, centipede are mixed to obtain animal medicinal materials, water extraction, separation to obtain filtrate and dregs; the remaining raw materials are mixed to obtain plant medicinal materials, CO2 supercritical extraction, collection of volatile oil and extraction residue; (3) the dregs and extraction residue of step (2) are mixed to obtain fermentation raw materials, anaerobic fermentation, sterilization after fermentation, filtration to obtain extract; (4) the filtrate of step (2) and the extract of step (3) are mixed, vacuum concentration to obtain clear paste, add volatile oil, stir evenly to obtain composite extraction product.

2. The combined extraction process of silkworm moth and Epimedium according to claim 1, characterized in that, The following ingredients are weighed according to the formula: Herba Epimedii 10-20 parts, Radix Morindae Officinalis 7-13 parts, Radix Rehmanniae Preparata 8-15 parts, silkworm moth 3-9 parts, Semen Leonuri 8-15 parts, Fructus Corni 9-17 parts, Eucommia ulmoides 3-9 parts, Angelica sinensis 6-12 parts, Fructus Lycii 7-14 parts, Cinnamomum cassia 2-9 parts, ginseng 1-10 parts, Gastrodia elata 3-11 parts, Cornu Cervi Pantotrichum 1-10 parts, Aconitum 4-10 parts, centipede 1-6 parts, Dioscorea opposita 10-17 parts, Cornu Cervi 1-8 parts.

3. The combined extraction process of silkworm moth and Epimedium according to claim 1, characterized in that, The male silkworm moth, Cornu Cervi, Cornu Cervi, centipede are mixed to obtain animal medicinal materials, crushed, soaked in water, reflux extraction for 1-2h, repeated extraction 2-4 times, filtration to combine the filtrate, and at the same time obtain the dregs.

4. The combined extraction process of silkworm moth and Epimedium according to claim 1, characterized in that, The remaining raw materials are mixed to obtain plant medicinal materials, crushed to less than 200 mesh, CO2 supercritical extraction, extraction pressure 30-35MPa, extraction temperature 40-50℃, extraction time 1-3h, collection of volatile oil and extraction residue.

5. The combined extraction process of silkworm moth and Epimedium according to claim 1, characterized in that, The conditions of anaerobic fermentation are: 35-38℃ anaerobic fermentation for 3-5 days.

6. The combined extraction process of silkworm moth and Epimedium according to claim 1, characterized in that, The strains include Lactobacillus coryniformis, Lactobacillus brevis and Lactobacillus rhamnosus.

7. The process for the combined extraction of silkworm moth and Epimedium herb as claimed in claim 6, wherein, The ratio of the number of viable bacteria of Lactobacillus coryniformis, Lactobacillus brevis and Lactobacillus rhamnosus in the strain is 1:(1.2-1.5):(0.6-0.8).

8. The combined extraction process of silkworm moth and Epimedium according to claim 1, characterized in that, The mass ratio of Herba Epimedii, Cornu Cervi and ginseng is (12-17):(2-6):(3-7).

9. The compound extraction process of silkworm moth and Epimedium according to claim 8, characterized in that, The following ingredients are weighed according to the formula: Herba Epimedii 15 parts, Radix Morindae Officinalis 10 parts, Radix Rehmanniae Preparata 12 parts, silkworm moth 8 parts, Semen Leonuri 10 parts, Fructus Corni 10 parts, Eucommia ulmoides 8 parts, Angelica sinensis 8 parts, Fructus Lycii 10 parts, Cinnamomum cassia 6 parts, ginseng 5 parts, Gastrodia elata 6 parts, Cornu Cervi Pantotrichum 4 parts, Aconitum 6 parts, centipede 3 parts, Dioscorea opposita 12 parts, Cornu Cervi 4 parts.

10. The application of the composite extraction process of silkworm moth and Herba Epimedii according to any one of claims 1-9 in the preparation of silkworm chit column Gouteng capsules.

Images