A fermented composition for tonifying the kidney and enhancing male virility and its preparation method

Abstract

This invention belongs to the field of traditional Chinese medicine fermentation technology, specifically relating to a kidney-tonifying and aphrodisiac fermentation composition and its preparation method. The composition, by weight, comprises 30-60 parts of Cistanche deserticola, 30-60 parts of black truffle, 10-30 parts of black beans, 5-20 parts of Eucommia ulmoides male flowers, 15-30 parts of Codonopsis pilosula, 5-15 parts of enzyme preparation, 0.6-1.4 parts of fermentation starter culture, and 800-900 parts of water. The composition, with Cistanche deserticola and black truffle as the main components, supplemented by black beans, Eucommia ulmoides male flowers, and Codonopsis pilosula, works synergistically to achieve a good kidney-tonifying and aphrodisiac effect after enzymatic fermentation.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine fermentation technology, specifically relating to a kidney-tonifying and aphrodisiac fermentation composition and its preparation method. Background Technology

[0002] Yang deficiency, a term in Traditional Chinese Medicine (TCM), refers to a pathological condition characterized by a decline in the body's Yang energy, resulting in reduced or weakened bodily functions, decreased metabolic activity, low reactivity, and insufficient Yang heat. Yang energy warms the limbs and internal organs; deficiency leads to impaired bodily function and a tendency towards coldness and weakness. Common types include stomach Yang deficiency, spleen Yang deficiency, and kidney Yang deficiency. Kidney Yang deficiency, also known as Mingmen fire deficiency, is characterized by symptoms such as lower back and knee pain and weakness, aversion to cold, cold limbs, pale or dark complexion, weakness, and low libido. In men, it manifests as impotence, spermatorrhea, and premature ejaculation; in women, it presents as uterine coldness and infertility, and excessive, thin, clear vaginal discharge. Kidney Yang deficiency is primarily influenced by lifestyle habits, constitution, age, and congenital factors. The common treatment principle for kidney Yang deficiency is to warm and tonify kidney Yang, including medication, massage, and health exercises. Commonly used formulas include Jin Kui Shen Qi Wan and You Gui Yin.

[0003] Chinese patent CN118058470A discloses a Cistanche deserticola and deer antler composition for tonifying the kidneys, strengthening yang, and improving bone health, and its preparation method. The composition includes 1000-3000 parts of a traditional Chinese medicine composition, 80-200 parts of a biological bacterial composition, 50-100 parts of an inorganic salt additive, 50-140 parts of a vitamin composition, 30-50 parts of edible salt, 10-20 parts of pectin, 10-20 parts of xanthan gum, 10-20 parts of polydextrose, 10-20 parts of sucralose, and 10-20 parts of potassium sorbate. The traditional Chinese medicine composition comprises the following raw materials in parts by weight: 50-200 parts of Cistanche deserticola, 350-400 parts of deer antler, 30-90 parts of Cornus officinalis, 30-90 parts of Eucommia ulmoides leaves, 30-90 parts of Dendrobium officinale, 30-70 parts of Astragalus membranaceus, and 30-50 parts of Poria cocos. 30-50 parts cloves, 30-90 parts yam, 30-200 parts ginseng, 20-90 parts silkworm pupae, 10-20 parts eucommia male flowers, 10-20 parts leeks, 10-20 parts white thorn fruit, 10-20 parts black rice, 10-20 parts sword beans, 10-20 parts black peanuts, 30-50 parts cinnamon, 30-50 parts longan pulp, 30-50 parts goji berries, 30-50 parts raspberries, 30-50 parts black soybeans, 30-50 parts alpinia oxyphylla, 50-200 parts dates, 10-20 parts walnut kernels, 10-20 parts raw oysters, 10-20 parts oysters, 10-20 parts osmanthus, 10-20 parts black sesame seeds, 10-20 parts mulberries. The ingredients include 10-20 parts of perilla seeds and 10-20 parts of black truffle. This patented method uses ultrasonic extraction of the traditional Chinese medicine, resulting in a low extraction rate and wasted resources. Summary of the Invention

[0004] To address the aforementioned technical problems, this invention provides a kidney-tonifying and aphrodisiac fermented composition and its preparation method. This composition utilizes an enzymatic fermentation process, which significantly improves the extraction rate of traditional Chinese medicine and enhances its kidney-tonifying and aphrodisiac effects.

[0005] In this invention, "bacterial activity", "bacterial viability", "live count", "live bacteria content", and "live bacteria quantity" have the same meaning under certain conditions.

[0006] In this invention, "enzyme activity", "enzyme activity", "enzyme activity" and "specific activity of enzyme" have the same meaning under certain conditions.

[0007] On the one hand, the present invention provides a fermented composition for tonifying the kidneys and enhancing male virility.

[0008] The fermented composition for tonifying the kidneys and strengthening yang, by weight, includes 30-60 parts of Cistanche deserticola, 30-60 parts of black truffle, 10-30 parts of black beans, 5-20 parts of Eucommia ulmoides male flowers, 15-30 parts of Codonopsis pilosula, 5-15 parts of enzyme preparation, 0.6-1.4 parts of fermentation starter, and 800-900 parts of water.

[0009] Preferably, the fermentation starter culture includes Lactobacillus plantarum and Bacillus subtilis.

[0010] More preferably, the weight ratio of the plant lactobacillus to the Bacillus subtilis is (2-5):(5-9).

[0011] More preferably, the bacterial activity of the *Lactobacillus plantarum* is 300-400 billion CFU / g, and the bacterial activity of the *Bacillus subtilis* is 200-300 billion CFU / g.

[0012] Preferably, the enzyme preparation includes cellulase and pectinase.

[0013] More preferably, the weight ratio of cellulase to pectinase is (1-5):(2-7).

[0014] The cellulase has an activity of 30,000-40,000 U / g, and the pectinase has an activity of 40,000-50,000 U / g.

[0015] Preferably, the weight ratio of the Cistanche deserticola to the black truffle is 1:1.

[0016] Preferably, the weight ratio of Cistanche deserticola to Codonopsis pilosula is 2.5:1.

[0017] More preferably, the weight ratio of Cistanche deserticola, black truffle, black bean, Eucommia ulmoides male flower and Codonopsis pilosula is 9:9:5:2:3.6.

[0018] Preferred, a kidney-tonifying and aphrodisiac composition, by weight, comprises 45 parts of Cistanche deserticola, 45 parts of black truffle, 25 parts of black soybean, 10 parts of Eucommia ulmoides male flower, 18 parts of Codonopsis pilosula, 3 parts of cellulase, 3 parts of pectinase, 0.3 parts of Lactobacillus plantarum, 0.6 parts of Bacillus subtilis, and 857 parts of water.

[0019] On the other hand, a method for preparing the above-mentioned kidney-tonifying and aphrodisiac fermented composition is provided, comprising the following steps:

[0020] (1) After passing the powders of Cistanche deserticola, black truffle, black beans, Eucommia ulmoides male flowers and Codonopsis pilosula through a 20-50 mesh sieve, add water and mix evenly. Adjust the pH to 5.0-6.0 to obtain a mixed solution.

[0021] (2) Add an enzyme preparation to the mixture for enzymatic hydrolysis. After the enzymatic hydrolysis is completed, adjust the pH to 6.5-7.0 to obtain the enzymatic hydrolysate.

[0022] (3) The enzyme hydrolysate is extracted at a temperature of 90-130℃ for 20-60 min to inactivate the enzyme and obtain the extract;

[0023] (4) Add fermentation bacteria powder to the extract and ferment to obtain fermentation liquid. The supernatant of the fermentation liquid after solid-liquid separation is fermentation liquid.

[0024] Preferably, the enzymatic hydrolysis temperature in step (2) is 30-60℃ and the enzymatic hydrolysis time is 60-100min.

[0025] Preferably, the fermentation temperature in step (4) is 20-30℃ and the fermentation time is 18-24h.

[0026] The technical solution of this invention has the following advantages:

[0027] 1. This invention uses Eucommia ulmoides male flowers, Cistanche deserticola, black beans, black truffles, and Codonopsis pilosula as raw materials. Eucommia ulmoides male flowers are the flowers of the male Eucommia ulmoides tree and contain more than 60 kinds of effective plant components. It can be said that Eucommia ulmoides male flowers possess all the health benefits of the Eucommia ulmoides plant, and are particularly effective in strengthening the liver, tonifying the kidneys, promoting bowel movements, inducing sleep, and lowering blood pressure, blood sugar, and cholesterol. The *Shennong Bencao Jing* records that Eucommia ulmoides is used to treat lower back pain, tonify the middle jiao (spleen and stomach), benefit essence and qi, strengthen tendons and bones, eliminate itching and dampness in the genital area, and treat dribbling urination. Long-term use can lighten the body and promote longevity. It is listed as a superior herb.

[0028] Cistanche deserticola is sweet and salty in taste, and warm in nature. It tonifies kidney yang, benefits essence and blood, and moistens the intestines. It is mainly used for impotence caused by kidney yang deficiency and insufficient essence and blood.

[0029] Black beans are the black seeds of the soybean plant, a legume. They contain 36% protein, which is easily digestible and plays a vital role in meeting the body's protein needs. They also contain 16% fat, primarily unsaturated fatty acids with an absorption rate as high as 95%. Besides meeting the body's fat requirements, black beans also help lower blood cholesterol. Furthermore, black beans are rich in vitamins, lecithin, melanin, and other substances, with particularly high levels of B vitamins and vitamin E, providing nutritional and health benefits. Black beans also contain abundant trace elements, essential for maintaining bodily functions, delaying aging, reducing blood viscosity, and meeting the brain's need for trace elements.

[0030] Codonopsis pilosula is sweet in taste and neutral in nature. It enters the spleen and lung meridians and has similar effects to ginseng, but its medicinal power is weaker. It has the effects of tonifying the middle and replenishing qi, quenching thirst, strengthening the spleen and benefiting the lungs, nourishing blood and promoting body fluids. It is often used for symptoms such as spleen and lung qi deficiency, poor appetite and fatigue, cough and wheezing, qi and blood deficiency, sallow complexion, palpitations and shortness of breath, thirst due to fluid depletion, and internal heat and thirst.

[0031] The above-mentioned ingredients are mainly Cistanche deserticola and black truffle, supplemented with black beans, Eucommia ulmoides male flowers and Codonopsis pilosula. After enzymatic hydrolysis and fermentation, they can improve anal temperature, free movement time, swimming time, testicular index and seminal vesicle index in mice with kidney yang deficiency, and have a very good effect of tonifying the kidney and strengthening yang.

[0032] 2. The kidney-tonifying and aphrodisiac composition provided by the present invention, when Cistanche deserticola, black truffle, black bean, Eucommia ulmoides male flower and Codonopsis pilosula are in a certain proportion, can significantly increase the anal temperature of mice in a mouse model of kidney yang deficiency and has a good relieving effect on symptoms of aversion to cold and cold limbs.

[0033] 3. The preparation method of the kidney-tonifying and aphrodisiac composition provided by the present invention involves adjusting the pH to 5.0-6.0 before enzymatic hydrolysis, under which the enzyme preparation can more easily exert its enzymatic hydrolysis effect. After enzymatic hydrolysis, the pH is adjusted to 6.5-7.0 to better exert the growth and decomposition ability of the strain, thereby facilitating the precipitation of small molecule substances and improving the kidney-tonifying and aphrodisiac efficacy. Detailed Implementation

[0034] To make the objectives, technical solutions, and advantages of this invention clearer, the embodiments of this invention will be described in further detail below. All other embodiments obtained by those skilled in the art based on the embodiments disclosed in this invention without inventive effort are within the scope of protection of this disclosure.

[0035] For any experimental steps or conditions not specified in the examples, the procedures or conditions described in the literature in this field can be followed. All reagents without specified manufacturers are commercially available standard reagent products.

[0036] In the following examples, the source information of some products is as follows: Bacillus subtilis was purchased from Shanghai Baiyi Biotechnology Co., Ltd., and the freeze-dried powder with product number BY-BC0069 had a bacterial activity of 300 billion CFU / g; Lactobacillus plantarum was purchased from Hebei Yiran Biotechnology Co., Ltd., and the freeze-dried powder with bacterial activity of 400 billion CFU / g; cellulase and pectinase were purchased from Shandong Longket Enzyme Preparation Co., Ltd., and the cellulase activity was 30,000 U / g and the pectinase activity was 40,000 U / g.

[0037] Example 1

[0038] This embodiment provides a composition for tonifying the kidney and strengthening yang, comprising 60g of Cistanche deserticola, 60g of black truffle, 10g of black beans, 20g of Eucommia ulmoides male flowers, 20g of Codonopsis pilosula, 830g of water, 1g of cellulase, 7g of pectinase, 0.2g of Lactobacillus plantarum and 0.9g of Bacillus subtilis.

[0039] The preparation method is as follows: (1) Crush 60g of Cistanche deserticola, 60g of black truffle, 10g of black beans, 20g of Eucommia ulmoides male flowers and 20g of Codonopsis pilosula into powder and pass them through a 30-mesh sieve. Then add 860g of water and mix evenly. Add sodium bicarbonate to adjust the pH to 5.0 to obtain a mixed solution. (2) Heat the mixed solution to 50℃, add 1g of cellulase and 7g of pectinase and hydrolyze for 70min. Add sodium bicarbonate to adjust the pH to 7.0 to obtain an enzymatic hydrolysate. (3) Transfer the enzymatic hydrolysate to a fermenter. Transfer the entire fermenter to a high-temperature sterilizer. Set the temperature of the high-temperature sterilizer to 90℃ to inactivate the enzyme and extract for 30min to obtain an extract. (4) Lower the temperature of the extract to 25℃, add 0.2g of Bacillus plantarum powder and 0.9g of Bacillus subtilis powder to the fermenter. Maintain the system temperature at 25℃ and ferment for 18h to obtain a fermentation liquid. The supernatant obtained by solid-liquid separation of the fermentation liquid is the fermentation clear liquid.

[0040] Example 2

[0041] This embodiment provides a composition for tonifying the kidney and strengthening yang, comprising 30g of Cistanche deserticola, 30g of black truffle, 30g of black beans, 5g of Eucommia ulmoides male flowers, 16g of Codonopsis pilosula, 88g of water, 5g of cellulase, 2g of pectinase, 0.5g of Lactobacillus plantarum and 0.5g of Bacillus subtilis.

[0042] The preparation method is as follows: (1) Cistanche deserticola 30g, black truffle 30g, black bean 30g, Eucommia ulmoides male flower 5g, Codonopsis pilosula 16g are crushed and passed through a 30-mesh sieve. Then, 889g of water is added and mixed evenly. Sodium bicarbonate is added to adjust the pH to 6.0 to obtain a mixed solution. (2) The mixed solution is heated to 30℃, 5g of cellulase and 2g of pectinase are added and enzymatically hydrolyzed for 100min. Sodium bicarbonate is added to adjust the pH to 7.0 to obtain an enzymatic hydrolysate. (3) The enzymatic hydrolysate is transferred to a fermenter. The entire fermenter is transferred to a high-temperature sterilizer. The temperature of the high-temperature sterilizer is set to 130℃ to inactivate the enzyme and extract for 20min to obtain an extract. (4) The temperature of the extract is lowered to 30℃. 0.5g of Bacillus plantarum powder and 0.5g of Bacillus subtilis powder are added to the fermenter. The system temperature is maintained at 30℃ and fermented for 20h to obtain a fermentation liquid. The supernatant obtained by solid-liquid separation of the fermentation liquid is the fermentation clear liquid.

[0043] Example 3

[0044] This embodiment provides a composition for tonifying the kidney and strengthening yang, comprising 50g of Cistanche deserticola, 50g of black truffle, 20g of black beans, 6g of Eucommia ulmoides male flowers, 24g of Codonopsis pilosula, 850g of water, 4g of cellulase, 5g of pectinase, 0.4g of Lactobacillus plantarum and 0.8g of Bacillus subtilis.

[0045] The preparation method is as follows: (1) Crush 50g of Cistanche deserticola, 50g of black truffle, 20g of black beans, 6g of Eucommia ulmoides male flowers and 24g of Codonopsis pilosula into powder and pass them through a 30-mesh sieve. Then add 850g of water and mix evenly. Add sodium bicarbonate to adjust the pH to 5.0 to obtain a mixed solution; (2) Heat the mixed solution to 60℃, add 4g of cellulase and 5g of pectinase and enzymatically hydrolyze for 60min. Add sodium bicarbonate to adjust the pH to 6.5 to obtain an enzymatic hydrolysate; (3) Transfer the enzymatic hydrolysate to a fermenter. Transfer the entire fermenter to a high-temperature sterilizer. Set the temperature of the high-temperature sterilizer to 100℃ to inactivate the enzyme and extract for 60min to obtain an extract; (4) Lower the temperature of the extract to 20℃, add 0.4g of Bacillus plantarum powder and 0.8g of Bacillus subtilis powder to the fermenter and maintain the system temperature at 20℃ for 24h to obtain a fermentation liquid. The supernatant obtained by solid-liquid separation of the fermentation liquid is the fermentation clear liquid.

[0046] Example 4

[0047] This embodiment provides a composition for tonifying the kidney and strengthening yang, comprising 45g of Cistanche deserticola, 45g of black truffle, 25g of black beans, 10g of Eucommia ulmoides male flowers, 18g of Codonopsis pilosula, 857g of water, 3g of cellulase, 3g of pectinase, 0.3g of Lactobacillus plantarum and 0.6g of Bacillus subtilis.

[0048] The preparation method is as follows: (1) Crush 45g of Cistanche deserticola, 45g of black truffle, 25g of black beans, 10g of Eucommia ulmoides male flowers and 18g of Codonopsis pilosula into powder and pass them through a 30-mesh sieve. Then add 857g of water and mix evenly. Add sodium bicarbonate to adjust the pH to 5.0 to obtain a mixed solution. (2) Heat the mixed solution to 40℃, add 3g of cellulase and 3g of pectinase and enzymatically hydrolyze for 80min. Add sodium bicarbonate to adjust the pH to 7.0 to obtain an enzymatic hydrolysate. (3) Transfer the enzymatic hydrolysate to a fermenter. Transfer the entire fermenter to a high-temperature sterilizer. Set the temperature of the high-temperature sterilizer to 110℃ to inactivate the enzyme and extract for 40min to obtain an extract. (4) Lower the temperature of the extract to 25℃, add 0.3g of Bacillus plantarum powder and 0.6g of Bacillus subtilis powder to the fermenter. Maintain the system temperature at 25℃ and ferment for 22h to obtain a fermentation liquid. The supernatant obtained by solid-liquid separation of the fermentation liquid is the fermentation clear liquid.

[0049] Comparative Example 1

[0050] This comparative example provides a kidney-tonifying and aphrodisiac composition, which differs from Example 4 in the proportion of Cistanche deserticola and black truffle. Specifically, it includes 85.71g of Cistanche deserticola, 4.29g of black truffle, 25g of black beans, 10g of Eucommia ulmoides male flowers, and 18g of Codonopsis pilosula. The rest is the same as in Example 4.

[0051] Comparative Example 2

[0052] This comparative example provides a kidney-tonifying and aphrodisiac composition, which differs from Example 4 in the proportion of Cistanche deserticola and Codonopsis pilosula. Specifically, it includes 18g of Cistanche deserticola, 45g of black truffle, 25g of black beans, 10g of Eucommia ulmoides male flowers, and 45g of Codonopsis pilosula. The rest is the same as in Example 4.

[0053] Comparative Example 3

[0054] This comparative example provides a kidney-tonifying and aphrodisiac composition, which differs from Example 4 in that it does not include Codonopsis pilosula. Specifically, it includes 51.48g of Cistanche deserticola, 51.48g of black truffle, 28.6g of black soybean, and 11.44g of Eucommia ulmoides male flowers. The rest is the same as in Example 4.

[0055] Comparative Example 4

[0056] This comparative example provides a kidney-tonifying and aphrodisiac composition, which differs from Example 4 in that Codonopsis pilosula is replaced with ginseng. Specifically, it includes 45g of Cistanche deserticola, 45g of black truffle, 25g of black soybean, 10g of Eucommia ulmoides male flowers, and 18g of ginseng. The rest is the same as in Example 4.

[0057] Comparative Example 5

[0058] This comparative example provides a kidney-tonifying and aphrodisiac composition, which differs from Example 4 in that it does not include black truffles. Specifically, it includes 65.67g of Cistanche deserticola, 0g of black truffles, 36.48g of black beans, 14.59g of Eucommia ulmoides male flowers, and 26.26g of Codonopsis pilosula. The rest is the same as in Example 4.

[0059] Comparative Example 6

[0060] This comparative example provides a kidney-tonifying and aphrodisiac composition, which differs from Example 4 in that the fermentation strain replaces Lactobacillus acidophilus with Bacillus subtilis, specifically including 0.3g of Lactobacillus plantarum and 0.6g of Lactobacillus acidophilus. The rest is the same as in Example 4.

[0061] Comparative Example 7

[0062] This comparative example provides a kidney-tonifying and aphrodisiac composition, which differs from Example 4 in that cellulase and pectinase are replaced with alkaline protease, i.e., the enzyme preparation is only 6g of alkaline protease. The rest is the same as in Example 4.

[0063] Comparative Example 8

[0064] This comparative example provides a kidney-tonifying and aphrodisiac composition comprising 45g of Cistanche deserticola, 45g of black truffle, 25g of black beans, 10g of Eucommia ulmoides male flowers, 18g of Codonopsis pilosula, 857g of water, 3g of cellulase, and 3g of pectinase.

[0065] The preparation method is as follows: (1) Crush 45g of Cistanche deserticola, 45g of black truffle, 25g of black beans, 10g of Eucommia ulmoides male flowers and 18g of Codonopsis pilosula into powder and pass them through a 30-mesh sieve. Then add 857g of water and mix evenly. Add sodium bicarbonate to adjust the pH to 5.0 to obtain a mixed solution; (2) Heat the mixed solution to 40℃, add 3g of cellulase and 3g of pectinase and hydrolyze for 80min. Add sodium bicarbonate to adjust the pH to 7.0 to obtain an enzymatic hydrolysate; (3) Transfer the enzymatic hydrolysate to a fermenter. Transfer the entire fermenter to a high-temperature sterilizer. Set the temperature of the high-temperature sterilizer to 110℃ and extract the enzyme for 40min to obtain an extract; (4) Separate the extract to obtain the supernatant.

[0066] Comparative Example 9

[0067] The composition powder was prepared according to the method of Example 1 in patent CN118058470A.

[0068] Experimental Example 1

[0069] 1. Sample preparation: The fermentation broths obtained in Examples 1-4 and Comparative Examples 1-7, and the supernatant obtained in Example 8 were spray-dried to obtain the composition powder.

[0070] 2. Modeling and Drug Administration

[0071] Healthy male ICR mice were randomly divided into four groups: a blank control group, a model group, Example 1 group, Example 2 group, Example 3 group, Example 4 group, and comparative examples 1, 2, 3, 4, 5, 6, 7, 8, and 9, with 10 mice in each group. Except for the blank control group, the other groups were intraperitoneally injected with a diluted 0.4 mg / mL estradiol benzoate soybean oil solution (0.1 mL / 10 g body weight) once daily for 15 days to establish a kidney-yang deficiency model. The blank control group was intraperitoneally injected with physiological saline (0.1 mL / 10 g body weight) for 15 days. Mouse characteristics were observed. Mice in the normal control group had smooth, dry fur, normal activity, and preferred movement to lying down. Mice in the other groups exhibited symptoms of kidney-yang deficiency, such as erect, sweaty, dull fur, lethargy, and a preference for warmth and aversion to cold, indicating successful model establishment.

[0072] After successful modeling, the mice in Examples 1-4 and Comparative Examples 1-9 were administered the corresponding composition powder by gavage at a dose of 260 mg / kg. The model group and the control group were administered the corresponding amount of distilled water by gavage once a day for 15 consecutive days.

[0073] 3. Indicator Testing

[0074] Rectal temperature was measured on day 14 of drug administration. Grasping force was measured on day 14 using a rat and mouse gripping force tester (model: YR22-YLS-13A), with three measurements taken for each mouse and the average value recorded. On day 14 of drug administration, mice were placed in an automated mouse activity tester (model: YLS-1C), and the number of mouse movements was recorded over 10 minutes. On day 15 of drug administration, the tail-bearing swimming time was measured. Mice were placed in water at 24±1℃, and sinking three times was considered an indicator of inability to swim; any time exceeding 1 hour was counted as 1 hour. After the weight-bearing swimming experiment, mice were euthanized by cervical dislocation, weighed, and the testes and seminal vesicles were removed. They were washed with physiological saline, and surface blood vessels, fat, connective tissue, etc., were removed. The moisture was absorbed with filter paper, and the mice were weighed using an electronic balance. The index (%) was calculated as: organ weight / body weight * 100%.

[0075] 4. Data Processing

[0076] Data are expressed as mean ± variance (X ± S). The t-test was used for comparisons between groups, and P < 0.05 was considered statistically significant.

[0077] 5. Data Analysis

[0078] 5.1 Influence of General Physical Signs

[0079] The control group mice had dry, smooth fur, were in good spirits, and exhibited normal activity levels, preferring to move rather than lie down. The model mice had sweaty, bristly fur, were lethargic, preferred warmth and cold, arched their backs, and preferred to huddle together rather than move, indicating successful modeling.

[0080] 5.2 The results of the effects on rectal temperature, spontaneous activity time, and swimming time are shown in Table 1.

[0081] Table 1 Effects of kidney-yang deficiency on anal temperature, spontaneous activity time, and swimming time in mice (x±s, n=10)

[0082]

[0083] Note: Compared with the same model group, * indicates P < 0.05, ** indicates P < 0.01; compared with Example 4, # indicates P < 0.05, ## indicates P < 0.01.

[0084] Kidney Yang deficiency syndrome includes major symptoms such as aversion to cold and cold limbs, decreased libido, and soreness and weakness of the lower back and knees, as well as minor symptoms such as lethargy, lower limb edema, pale tongue with white coating, and weakness in the cun position. In this experiment, rectal temperature was used to reflect the symptoms of aversion to cold and cold limbs. As shown in Table 1, the model group showed a highly significant difference compared to the blank control group, indicating that the model of aversion to cold and cold limbs was successfully established. Examples 1-4 showed significant differences compared to the model control group, indicating that the fermented composition provided by this invention can effectively alleviate the symptoms of aversion to cold and cold limbs in a mouse model of kidney Yang deficiency. The comparative group showed a significant difference compared to Example 4, meaning that the five raw materials selected in this invention—Cistanche deserticola, black truffle, black bean, Eucommia ulmoides, and Codonopsis pilosula—exhibited their best effects after enzymatic fermentation.

[0085] In this experiment, the number of spontaneous activities was used to reflect the symptoms of lethargy. As shown in Table 1, the model group showed a highly significant difference compared with the blank control group, indicating that the model of lethargy was successfully established. Examples 1-4 showed highly significant differences compared with the model control group, indicating that the fermentation composition provided by this invention can effectively alleviate the symptoms of lethargy in the mouse model of kidney yang deficiency. Although some comparative examples showed significant differences compared with the model group, all comparative examples showed significant differences compared with Example 4. This means that although some comparative examples can alleviate the symptoms of lethargy in the mouse model of kidney yang deficiency, all comparative examples showed significant differences compared with Example 4. This means that the five raw materials selected in this invention—Cistanche deserticola, black truffle, black bean, Eucommia ulmoides, and Codonopsis pilosula—only exert their better effects after enzymatic fermentation.

[0086] In this experiment, the duration of weight-bearing swimming was used to reflect the symptoms of lower back and knee weakness. Table 1 shows that the model group had a highly significant difference compared to the blank control group, indicating that the lower back and knee weakness symptom model was successfully established. Groups 1-4 of the present invention showed highly significant differences compared to the model group, indicating that the fermented composition provided by this invention can effectively improve the symptoms of lower back and knee weakness in a mouse model of kidney yang deficiency. The comparative group showed a significant difference compared to Example 4, meaning that the five raw materials selected in this invention—Cistanche deserticola, black truffle, black bean, Eucommia ulmoides, and Codonopsis pilosula—exhibited their best effects after enzymatic fermentation.

[0087] 5.3 The effects on the testicular index and seminal vesicle index of mice with kidney yang deficiency are shown in Table 2.

[0088] Table 2. Effects of Kidney Yang Deficiency on Testicular Index and Seminal Vesicle Index (x±s, n=10)

[0089] Note: Compared with the same model group, * indicates P < 0.05, ** indicates P < 0.01; compared with Example 4, # indicates P < 0.05, ## indicates P < 0.01.

[0090] In this experiment, the testicular index and seminal vesicle index were used to reflect the symptoms of decreased libido in a mouse model of kidney-yang deficiency. Table 2 shows that compared with the model control group, the testicular index and seminal vesicle index of Examples 1-4 were significantly different, while there was no significant difference compared with the blank control group, indicating that the fermentation composition provided by this invention can improve libido in mice. Furthermore, the comparative groups also showed significant differences compared with Example 4 group, meaning that the raw materials, strains, and enzymes selected in this invention work together to achieve a more effective result.

[0091] In summary, the fermentation composition provided by this invention can improve symptoms such as soreness and weakness of the lower back and knees, aversion to cold and cold limbs, lethargy, and decreased libido in a mouse model of kidney yang deficiency.

[0092] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, and is not intended to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions made by those skilled in the art to the technical solution of the present invention do not depart from the essence and scope of the technical solution of the present invention.

Claims

1. A fermented composition for tonifying the kidneys and enhancing male virility, characterized in that, By weight, it consists of 30-60 parts of Cistanche deserticola, 30-60 parts of black truffle, 10-30 parts of black beans, 5-20 parts of Eucommia ulmoides male flowers, 15-30 parts of Codonopsis pilosula, 5-15 parts of enzyme preparation, 0.6-1.4 parts of fermentation starter powder and 800-900 parts of water. The fermentation starter culture is Lactobacillus plantarum and Bacillus subtilis; the weight ratio of Lactobacillus plantarum and Bacillus subtilis is (2-5):(5-9). The enzyme preparation is cellulase and pectinase; the weight ratio of cellulase to pectinase is (1-5):(2-7).

2. The composition according to claim 1, characterized in that, The weight ratio of the Cistanche deserticola to the black truffle is 1:

1.

3. The composition according to claim 1, characterized in that, The weight ratio of the Cistanche deserticola to the Codonopsis pilosula is 2.5:

1.

4. The composition according to claim 1, characterized in that, The weight ratio of Cistanche deserticola, black truffle, black bean, Eucommia ulmoides male flower and Codonopsis pilosula is 9:9:5:2:3.

6.

5. The composition according to claim 4, characterized in that, By weight, it consists of 45 parts of Cistanche deserticola, 45 parts of black truffle, 25 parts of black beans, 10 parts of Eucommia ulmoides male flowers, 18 parts of Codonopsis pilosula, 3 parts of cellulase, 3 parts of pectinase, 0.3 parts of Lactobacillus plantarum, 0.6 parts of Bacillus subtilis, and 857 parts of water.

6. A method for preparing the kidney-tonifying and aphrodisiac composition according to any one of claims 1-5, comprising the following steps: (1) After passing the powders of Cistanche deserticola, black truffle, black beans, Eucommia ulmoides male flowers and Codonopsis pilosula through a 20-50 mesh sieve, add water and mix evenly. Adjust the pH to 5.0-6.0 to obtain a mixed solution. (2) Add an enzyme preparation to the mixture for enzymatic hydrolysis. After the enzymatic hydrolysis is completed, adjust the pH to 6.5-7.0 to obtain the enzymatic hydrolysate. (3) The enzyme hydrolysate is extracted at a temperature of 90-130℃ for 20-60 min to inactivate the enzyme and obtain the extract; (4) Add fermentation bacteria powder to the extract and ferment to obtain fermentation liquid. The supernatant of the fermentation liquid after solid-liquid separation is fermentation liquid. in, In step (2), the enzymatic hydrolysis temperature is 30-60℃ and the enzymatic hydrolysis time is 60-100min; in step (4), the fermentation temperature is 20-30℃ and the fermentation time is 18-24h.

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