Traditional Chinese medicine composition for nourishing and regulating kidney functions

Abstract

The invention discloses a traditional Chinese medicine composition for nourishing and regulating kidney functions. The traditional Chinese medicine composition is prepared by adopting a graded extraction and polypeptide composite technology. Herba epimedii, radix morindae officinalis, ginseng and eucommia ulmoides male flowers are used as main medicinal materials, and active ingredient concentrated solutions are respectively obtained through an ethanol graded extraction process and a dynamic countercurrent extraction process. Meanwhile, small molecule peptides such as rhizoma polygonati peptide, puerarin peptide and oyster peptide are integrated, a stable micelle system is formed through step-by-step mixing, and finally the stable micelle system and the maca extract are compounded and homogenized. According to the process, the extraction process is optimized, effective components are reserved to the maximum extent, and the bioavailability is improved. The obtained composition has the advantages of traditional Chinese medicine multi-target regulation and peptide nutritional supplement, can warm and tonify kidney yang, improve kidney microcirculation and enhance the metabolic function, and is suitable for hypofunction caused by kidney deficiency. The product is stable in component and efficient in absorption, and has a remarkable effect in the aspect of kidney function regulation.

Description

Technical Field

[0001] The present invention belongs to the technical field of traditional Chinese medicine compositions, and particularly relates to a traditional Chinese medicine composition for nourishing and regulating kidney function. Background Art

[0002] Epimedium brevicornu is a perennial herb of the Berberidaceae family. It was first recorded in "Shennong Ben Cao Jing" and has a medicinal history of more than 2,000 years. Its stems and leaves contain active ingredients such as icariin and flavonoid compounds, and are traditionally used for tonifying the kidney and strengthening yang, expelling wind and removing dampness. Icariin can promote the secretion of sex hormones, enhance the function of the hypothalamus-pituitary-gonadal axis, and improve sexual function; flavonoid components protect kidney cells through antioxidant and anti-inflammatory effects, and relieve soreness in the waist and knees and impotence caused by kidney deficiency. Epimedium brevicornu has both the effects of tonifying the kidney and anti-fatigue, and has a significant effect on sexual dysfunction of the kidney-yang deficiency type, and can also regulate the immune system function.

[0003] Morinda officinalis is a vine shrub of the Rubiaceae family, mainly produced in southern China, and is one of the "Four Major Southern Medicines". Its roots contain components such as anthraquinones and nystose, and belong to the kidney and liver meridians. It is traditionally used for tonifying kidney yang and strengthening bones and muscles. The nystose and anthraquinone substances in it can enhance the function of the adrenal cortex, promote testosterone secretion, and improve impotence and infertility due to kidney-yang deficiency; its polysaccharide component can reduce kidney damage by inhibiting inflammatory factors. Morinda officinalis is warm but not dry, and long-term use can improve reproductive function and rheumatism pain. It is included in the longevity diet.

[0004] Panax ginseng is the rhizome of a plant of the Araliaceae family, containing active ingredients such as ginsenosides and polysaccharides, and is known as the "king of all herbs". "Shennong Ben Cao Jing" says that it "tonifies the five internal organs and calms the mind", and belongs to the spleen, lung, and kidney meridians. Ginsenoside Rg1 in it enhances adrenal function by regulating the HPA axis and improves fatigue and low immunity caused by insufficient kidney qi; its antioxidant effect can slow down oxidative damage to the kidneys. Ginseng can bidirectionally regulate immune function and has a significant effect on those with kidney deficiency accompanied by qi deficiency and the chronically ill and weak, but it should be used with caution in cases of excessive heat syndrome.

[0005] Oyster peptide is a small molecule peptide extracted from oysters by enzymatic hydrolysis, retaining components such as zinc and taurine. Traditional Chinese medicine believes that oysters are salty and cold and enter the kidney, and can nourish yin and suppress yang. The zinc element in it directly promotes testosterone synthesis, and the peptide substance enhances mitochondrial function by activating the AMPK signaling pathway, improving sexual function decline and fatigue of the kidney-yin deficiency type. Oyster peptide has high bioavailability and also has the effects of protecting the liver and lowering blood sugar, and is suitable for conditioning sub-health in modern people.

[0006] Eucommia male flowers are the male flower buds of Eucommiaceae, containing components such as chlorogenic acid and aucubin. "Shennong Ben Cao Jing" states that it "tonifies the middle-jiao and benefits essence-qi", and it is known as the "plant gold". Among them, aucubin reduces kidney inflammation by inhibiting the NF-κB pathway, and chlorogenic acid regulates lipid metabolism to improve renal hypertension; its high potassium and low sodium characteristics help maintain electrolyte balance. Eucommia male flowers have both antihypertensive and anti-aging effects, and are beneficial to patients with kidney deficiency accompanied by osteoporosis and metabolic syndrome.

[0007] Cordyceps militaris is a fungus of the family Clavicipitaceae, containing components such as cordycepin and cordycepic acid. Traditional Chinese medicine believes that it "nourishes both yin and yang", can benefit the lungs and kidneys, and stop bleeding and resolve phlegm. Among them, cordycepin reduces glomerular inflammation by regulating the Th1 / Th2 immune balance; cordyceps polysaccharide promotes DNA repair of kidney cells and improves proteinuria in chronic kidney disease. The quality of artificially cultivated Cordyceps militaris is stable, and it has a significant effect on patients with kidney deficiency accompanied by cough and asthma and low immunity.

[0008] Maca (Lepidium meyenii) is a traditional Peruvian medicinal material of the family Brassicaceae, containing alkaloids such as macamides and macaenes. Its dried roots are used to enhance sexual function and anti-fatigue. Among them, macamides improve sexual desire by regulating dopamine secretion in the hypothalamus, and its sterol components can protect Leydig cells in the testis and relieve infertility caused by kidney essence deficiency.

[0009] Polygonatum sibiricum peptide is enzymatically extracted from Polygonatum sibiricum, containing small molecule peptides (<800 Da). Polygonatum sibiricum has traditionally been used to nourish yin and moisten the lungs, and tonify the spleen and kidneys. Among them, the peptide substances reduce renal oxidative stress by activating the Nrf2 / ARE pathway, and its polysaccharide components regulate the Th17 / Treg balance to improve diabetic nephropathy. Polygonatum sibiricum peptide is suitable for patients with kidney yin deficiency type diabetes and menopausal syndrome, and has synergistic effects of protecting the liver and anti-aging.

[0010] Lycium barbarum peptide is extracted from Lycium barbarum in Ningxia, containing glycopeptide complex (LbGP). "Compendium of Materia Medica" states that Lycium barbarum "nourishes the kidneys and moistens the lungs", and it is an excellent product that is both a medicine and a food. Among them, Lycium barbarum glycopeptide reduces tubulointerstitial fibrosis by inhibiting the TLR4 / MyD88 pathway, and its antioxidant components protect the glomerular basement membrane.

[0011] Pueraria lobata peptide is derived from the rhizome of Pueraria lobata, containing components such as isoflavone aglycones. Traditional Chinese medicine believes that it relieves muscle stiffness and fever, promotes the production of body fluid and quenches thirst, and belongs to the lung and stomach meridians. Among them, puerarin reduces the damage of diabetic nephropathy by inhibiting the generation of AGEs, and its anti-inflammatory effect improves glomerulosclerosis. Pueraria lobata peptide is suitable for patients with kidney yin deficiency accompanied by hypertension and menopausal hot flashes, and has a synergistic effect of protecting the cardiovascular system.

[0012] Soybean peptides are the proteolytic products of soybeans and contain oligopeptides. Their easy absorption characteristics have classified them as specific health foods in Japan. The soybean peptides in them regulate blood pressure by inhibiting ACE activity, and their antioxidant peptides reduce heavy metal-induced nephrotoxicity.

[0013] L-arginine is a conditionally essential amino acid and participates in the synthesis of nitric oxide. Its hydrochloride is used for hepatic encephalopathy and immunomodulation. The NO-mediated vasodilation among them improves renal blood flow and promotes the urea cycle to reduce blood ammonia; its effect of enhancing sperm motility is beneficial to male infertility.

[0014] Traditional Chinese medicine theory believes that the kidney is the foundation of congenital constitution, and kidney deficiency can lead to various diseases such as sexual dysfunction, decreased immunity, and metabolic disorders. The synergistic application of Chinese medicinal herbs for tonifying kidney yang such as Epimedium and Morinda officinalis and qi-tonifying and yin-nourishing components such as ginseng and oyster peptides can bidirectionally regulate the function of the hypothalamic-pituitary-target gland axis and improve kidney deficiency symptoms through multi-target effects. Modern research has confirmed that components such as flavonoids, saponins, and bioactive peptides in Chinese medicinal materials have antioxidant, anti-inflammatory, and cytoprotective effects, providing a scientific basis for the conditioning of renal function.

[0015] With the expansion of the sub-healthy population and the advent of an aging society, traditional Chinese medicine compositions with both traditional efficacy and modern technological advantages show broad application prospects. However, direct mixing of small molecule peptides and plant extracts is prone to the Maillard reaction, resulting in browning of active ingredients and loss of activity. How to integrate the small molecule peptide and traditional Chinese medicine extraction processes and study a natural and effective traditional Chinese medicine composition for improving the nourishing and conditioning of renal function is the research focus of the present invention. Summary of the Invention

[0016] The purpose of the present invention is to provide a traditional Chinese medicine composition for nourishing and conditioning renal function.

[0017] To solve the above technical problems, the specific process of the present invention is as follows:

[0018] Ethanol fractional extraction: First-stage extraction: Add 20 times the ethanol to 18 - 20 parts of Epimedium crushed to 80 mesh according to the mass ratio, reflux at 60 - 65°C for 40 min to obtain filtrate A and residue A; Second-stage extraction: Add 15 times the mass fraction of 50% ethanol to the residue A and 12 - 15 parts of Morinda officinalis crushed to 80 mesh according to the mass ratio, extract at 45°C with an ultrasonic frequency of 40 kHz for 30 min to obtain filtrate B and residue B; Combine filtrates A and B, and perform vacuum concentration at a pressure of -0.08 MPa and a pulse frequency of 0.5 Hz, with a temperature of 60°C, and concentrate to a relative density of 1.25 to obtain the Epimedium-Morinda officinalis concentrated solution;

[0019] Add 5 - 7 parts of Cordyceps militaris peptide and 10 - 15 parts of L - arginine to 50 parts of water, control the liquid temperature at 35°C, and use a high - speed shear emulsifier to disperse at 3000 rpm for 15 min until completely dissolved to obtain a mixed solution of Cordyceps militaris peptide and L - arginine;

[0020] Compound and drying: Preheat the Epimedium - Morinda officinalis concentrated liquid to 35 - 40°C, start magnetic stirring at 300 rpm, after stirring for 5 min, add the Panax ginseng - Eucommia ulmoides male flower concentrated liquid, raise the temperature to 45°C, stir at a constant temperature for 10 min, then cool down to 35°C, add the homogenized peptide micelles at a flow rate of 10 mL / min, switch to a homogenizer at 8000 rpm for 3 min, then add the above - mentioned mixed solution of Cordyceps militaris peptide and L - arginine at a flow rate of 5 mL / min, mix at a low speed of 200 rpm at room temperature for 20 min, then add 8 - 10 parts of Maca extract, raise the temperature to 35°C, stir at 400 rpm for 15 min, and finally place it in an 80°C environment for drying for 30 min.

[0021] Among them, the mass fraction of ethanol used in the first - stage extraction of ethanol fractional extraction is 70%, and the mass fraction of ethanol used in the second - stage extraction is 50%.

[0022] Among them, the Panax ginseng - Eucommia ulmoides male flower concentrated liquid includes dynamic counter - current extraction: Add 15 - 18 parts of sliced Panax ginseng and 8 - 12 parts of Eucommia ulmoides male flower crushed to 60 meshes to 25 times the mass fraction of 60% ethanol solution according to the mass ratio, perform dynamic counter - current extraction at 40°C, with a flow rate of 1.2 L / min for 2 h, and recover ethanol by rotary evaporation of the filtrate at 50°C to obtain the Panax ginseng - Eucommia ulmoides male flower concentrated liquid.

[0023] Among them, the sliced Panax ginseng also includes the following treatment: Prepare Panax ginseng, cut it into sliced Panax ginseng with a thickness of 2 mm, add 0.05% pectinase and 50 parts of water, and pre - treat and soak at 30°C for 20 min.

[0024] Among them, the homogenized peptide micelles include: Take 8 - 10 parts of Polygonatum sibiricum peptide and 5 - 6 parts of Pueraria lobata peptide and add them to the reaction kettle, add 80 parts of water according to the mass ratio, stir at a low speed of 60 rpm at 50°C for 30 min to form a colloidal matrix, slowly add 12 parts of soybean peptide substances to the colloidal matrix, sprinkle them evenly in 3 times, with an interval of 5 min each time, maintain the temperature at 50°C, increase the stirring speed to 80 rpm, and continuously mix for 40 min to ensure that low - molecular - weight peptides fully fill the pores of the colloid; Add 10 - 12 parts of Ostrea gigas peptide and 6 - 8 parts of Lycium barbarum peptide to 100 parts of water, a premixed solution stirred at 200 rpm for 10 min, add it in two times, with an interval of 10 min each time, and finally add L - arginine to adjust the pH to 6.8, stir at 60 rpm at 50°C for 2 h to form homogenized peptide micelles.

[0025] The beneficial effects of the present invention are as follows:

[0026] 1. The present invention relates to a traditional Chinese medicine composition for nourishing and conditioning kidney function. By integrating multiple traditional Chinese medicine active ingredients through a multi-step process, the synergistic effect of the medicinal efficacy is significantly enhanced. The composition takes epimedium, morinda officinalis, ginseng, and eucommia male flowers as the core medicinal materials, combines small molecule peptides such as polygonatum peptide, pueraria peptide, and oyster peptide, and is prepared by ethanol fractionation extraction, dynamic countercurrent extraction, and polypeptide mixing technology. Epimedium and morinda officinalis are extracted and concentrated by two-stage ethanol extraction to retain flavonoid active ingredients; ginseng and eucommia male flowers are subjected to enzymatic hydrolysis-assisted dynamic countercurrent extraction to improve the dissolution rate of saponins; various peptides form a micelle structure through stepwise mixing to enhance bioavailability. The final complex is homogenized, emulsified, and freeze-dried at low temperature to ensure the stability of the components. The advantages of this process are as follows: the extraction efficiency is optimized by fractionated extraction according to the characteristics of different medicinal materials; secondly, peptides synergize with traditional Chinese medicine components to enhance the antioxidant and energy metabolism functions of the kidney, and low-temperature treatment protects the activity of heat-sensitive components. The obtained composition has multiple effects such as warming the kidney yang, improving renal microcirculation, and regulating endocrine, and is applicable to symptoms such as fatigue and hypofunction caused by kidney deficiency. Moreover, the preparation has good uniformity and high absorption rate, providing an efficient and safe process formula for kidney function conditioning.

[0027] 2. The gradient ethanol dynamic extraction technology adjusts the ethanol concentration gradient to precisely match the polarity differences between epimedium flavonoid glycosides and morinda officinalis iridoids, simultaneously achieving the efficient extraction of lipophilic and hydrophilic components and avoiding the limitations of single-solvent extraction. Combining the synergistic effect of microwave-ultrasound, it quickly breaks the cell wall to release intracellular active substances, enhancing the dissolution efficiency. At the same time, low-temperature countercurrent extraction and pulsed vacuum concentration avoid the destruction of heat-sensitive components such as ginsenoside Rg1 and eucommia geniposide by high temperature, ensuring the integrity and biological activity of key components such as icariin and iridoids, providing a material basis for the multi-pathway mechanism of kidney protection.

[0028] 3. Dynamic countercurrent extraction and continuous phase change effect significantly shorten the extraction time, with significantly improved efficiency compared to traditional processes, and high ethanol recycling rate, reducing solvent consumption. Pulsed vacuum concentration destroys the surface tension of the solution through pressure oscillation, and the evaporation efficiency is higher than that of conventional vacuum concentration. The low-temperature condition synchronously protects the component stability. Pectinase pretreatment can soften the cell wall of ginseng, reduce the extraction energy consumption, and combine with the stage-by-stage penetration of gradient ethanol to achieve the integrated extraction of multi-polar components such as flavonoids, saponins, and phenylpropanoids, improving the active ingredients and simplifying the process steps, meeting the requirements of green manufacturing.

[0029] 4. The functional synergy between epimedium-morinda officinalis and ginseng-eucommia male flowers. Icariin can improve renal blood flow, morinda officinalis iridoids inhibit fibrosis, ginsenoside Rg1 resists renal tubular apoptosis, and eucommia geniposide enhances antioxidant activity, forming a multi-target kidney protection network. Through the scientific compatibility of ethanol gradient and low-temperature technology, the quality of the preparation is guaranteed.

[0030] 5. At 50 °C and pH 6.8, polygonatum peptide and pueraria peptide polysaccharide self-assemble through hydrogen bonds to form a three-dimensional colloidal network. Soybean peptide fills the pores in three intervals, reducing the system viscosity through physical embedding and dispersing low-molecular-weight peptides, avoiding the problems of bitter peptide residues and broad molecular weight distribution caused by traditional enzymatic hydrolysis processes. The colloidal network blocks the contact between free amino groups and reducing sugars through steric hindrance effects, inhibiting browning caused by the Maillard reaction and the oxidation inactivation of active peptides. At the same time, hydrophobic oyster peptide and wolfberry peptide are directionally adsorbed on the surface of the micelles, forming a "core-shell" structure, further shielding oxidation-sensitive groups and retaining the synergistic antioxidant activity of zinc-chelating peptide and wolfberry polysaccharide.

[0031] 6. By combining staged mixing with temperature gradient control and achieving hierarchical integration of peptide chains through shear force differences: polygonatum peptide first forms a framework with pueraria peptide, soybean peptide fills the pores, and oyster peptide and wolfberry peptide are adsorbed on the outer layer, forming micelles with multi-level pore sizes, which target and match intestinal peptide transport carriers, greatly improving the absorption rate compared to traditional mixing processes. L-arginine adjusts the pH to 6.8 in stages to stabilize the zeta potential of the micelles, avoiding aggregation caused by reaching the isoelectric point and ensuring the homogeneity and storage stability of the system.

[0032] 7. This process activates the active ingredients of different medicinal materials step by step through staged gradient mixing and precise temperature control technology: first, magnetic stirring is used to activate lipophilic substances, and then a colloidal network is formed by controlling the temperature; gradient cooling is combined with low-speed injection of peptide complexes and high-speed stirring to achieve directional embedding; nanocarriers are prepared by homogenization shearing while retaining the activity of small-molecule peptides; finally, yang-tonifying ingredients are integrated and the structure is quickly dehydrated and solidified. This technology breaks through the limitations of traditional processes, enhances the renal function conditioning effect significantly through the synergistic action of ingredients while improving the bioavailability, and has the advantages of active protection and process efficiency. Specific implementation mode

[0033] The following further elaborates on the present invention in conjunction with embodiments. The neutral protease used in the following implementation modes is Novozymes The product specification of flavor protease: 30,000 u / g, the execution standard: GB1886.174, the model of alkaline protease is Novozymes Alcalase 2.4L FG, and the model of glucoamylase is Novozymes Dextrozyme GA. All of the above enzymes are purchased from Beijing Gaoruisen Technology Co., Ltd. The preparation of all polypeptide substances includes:

[0034] Oyster peptide: After washing, mashing and homogenizing oyster meat, adjust the pH to 7.0, add 0.5% neutral protease according to the mass of the substrate, hydrolyze at 50 °C for 2 hours, then add 0.3% flavor protease. Hydrolyze at 55 °C for 1 hour, quickly raise the temperature to 85 °C, keep it for 5 min to inactivate the enzyme, and then centrifuge to take the supernatant, add 2% food-grade activated carbon and stir for 30 min to remove fishy smell, and then filter and concentrate.

[0035] Soybean peptide: Adjust the pH of soy protein isolate to 8.0, add 0.4% alkaline protease, hydrolyze at 55°C for 3 hours, and perform ultrafiltration after inactivating the enzyme.

[0036] Polygonatum sibiricum peptide: Add 5 times the amount of water to Polygonatum sibiricum powder, adjust the pH to 6.5, add 0.6% complex enzyme, the complex enzyme is cellulase: papain = 1:2, hydrolyze at 50°C for 4 hours, and perform nanofiltration and concentration after inactivating the enzyme.

[0037] Pueraria lobata peptide: Adjust the pH of Pueraria lobata starch residue to 5.0, add 0.4% glucoamylase, pretreat at 60°C for 1 hour, then add 0.5% trypsin, pH 7.5, and hydrolyze at 45°C for 3 hours.

[0038] Lycium barbarum peptide: Grind Lycium barbarum to 60 mesh, add water according to the solid-liquid ratio of 1:10, adjust the pH to 8.0, add 0.4% alkaline protease, hydrolyze at 50°C for 3 hours, then adjust the pH to 6.5, add 0.2% flavor protease, continue to hydrolyze for 1 hour, keep at 85°C for 10 minutes to inactivate the enzyme, perform ultrafiltration with a cut-off molecular weight of 1 kDa, and collect small molecular peptide segments.

[0039] Example 1

[0040] (1) Ethanol fractional extraction: First-stage extraction: Add 20 times the mass fraction of 70% ethanol to 19 parts of Epimedium brevicornu powder ground to 80 mesh according to the mass ratio, reflux at 62°C for 40 min to obtain filtrate A and residue A; Second-stage extraction: Add 15 times the mass fraction of 50% ethanol to residue A and 13 parts of Morinda officinalis powder ground to 80 mesh according to the mass ratio, extract at 45°C with ultrasound at 40 kHz for 30 min to obtain filtrate B and residue B; Combine filtrates A and B, perform vacuum concentration at a pressure of -0.08 MPa and a pulse frequency of 0.5 Hz, with a temperature of 60°C, and concentrate to a relative density of 1.25 to obtain the Epimedium brevicornu - Morinda officinalis concentrated solution;

[0041] (2) Dynamic countercurrent extraction: Prepare 16 parts of ginseng cut into slices with a thickness of 2 mm, add 0.05% pectinase, 50 parts of water, pretreat and soak at 30°C for 20 min, then add 25 times the mass fraction of 60% ethanol solution to it and 10 parts of Eucommia ulmoides male flower powder ground to 60 mesh according to the mass ratio, perform dynamic countercurrent extraction at 40°C, with a flow rate of 1.2 L / min for 2 h, and recover ethanol by rotary evaporation of the filtrate at 50°C to obtain the ginseng - Eucommia ulmoides male flower concentrated solution;

[0042] (3) Polypeptide mixing: Take 9 parts of polygonatum sibiricum peptide and 5.5 parts of pueraria lobata peptide and add them to the reaction kettle. Add 80 parts of water according to the mass ratio. At 50 °C, stir at a low speed of 60 rpm for 30 min to form a colloidal matrix. Slowly add 12 parts of soybean peptide substance to the colloidal matrix, sprinkle it evenly in 3 times, with an interval of 5 min each time. Maintain the temperature at 50 °C, increase the stirring speed to 80 rpm, and continue mixing for 40 min to ensure that low-molecular-weight peptides fully fill the pores of the colloid; Add 11 parts of oyster peptide and 7 parts of wolfberry peptide to 100 parts of water, a premixed solution stirred at 200 rpm for 10 min, add it in two times, with an interval of 10 min each time. Finally, add L-arginine to adjust the pH to 6.8, stir at 50 °C and 60 rpm at a low speed for 2 h to form a homogeneous peptide micelle; Add 6 parts of cordyceps militaris peptide and 13 parts of L-arginine to 50 parts of water, control the liquid temperature at 35 °C, use a high-speed shear emulsifier, disperse at 3000 rpm for 15 min until completely dissolved to obtain a mixed solution of cordyceps militaris peptide and L-arginine;

[0043] (4) Composite and drying: Preheat the epimedium-morinda officinalis concentrated liquid to 38 °C, turn on the magnetic stirrer at 300 rpm, stir for 5 min, then add the ginseng-epimedium pubescens concentrated liquid, heat up to 45 °C, stir at a constant temperature for 10 min, then cool down to 35 °C, add the above homogeneous peptide micelle at a flow rate of 10 mL / min, switch to a homogenizer at 8000 rpm for 3 min, then add the above mixed solution of cordyceps militaris peptide and L-arginine at a flow rate of 5 mL / min, mix at a low speed of 200 rpm at room temperature for 20 min, then add 9 parts of maca extract, heat up to 35 °C, stir at 400 rpm for 15 min, and finally dry in an 80 °C environment for 30 min.

[0044] Example 2

[0045] (1) Ethanol fractional extraction: Primary extraction: Crush 20 parts of epimedium to 80 mesh, add 20 times the mass fraction of 70% ethanol according to the mass ratio, reflux at 60 °C for 40 min to obtain filtrate A and residue A; Secondary extraction: Add residue A and 15 parts of morinda officinalis crushed to 80 mesh, supplement 15 times the mass fraction of 50% ethanol according to the mass ratio, extract at 45 °C with ultrasonic wave at 40 kHz for 30 min to obtain filtrate B and residue B; Combine filtrates A and B, carry out vacuum concentration at a pressure of -0.08 MPa and a pulse frequency of 0.5 Hz, with the temperature at 60 °C, concentrate to a relative density of 1.25 to obtain the epimedium-morinda officinalis concentrated liquid;

[0046] (2) Dynamic countercurrent extraction: Prepare 15 portions of ginseng sliced into pieces with a thickness of 2 mm, add 0.05% pectinase and 50 portions of water, pretreat and soak at 30 °C for 20 min, then add 12 portions of Eucommia ulmoides male flowers crushed to 60 mesh according to the mass ratio and add 25 times the mass fraction of 60% ethanol solution, perform dynamic countercurrent extraction at 40 °C with a flow rate of 1.2 L / min for 2 h, and rotate and evaporate the filtrate at 50 °C to recover ethanol to obtain the ginseng-Eucommia ulmoides male flower concentrate;

[0047] (3) Polypeptide mixing: Take 8 portions of polygonatum sibiricum peptide and 6 portions of pueraria lobata peptide and add them to the reaction kettle, add 80 portions of water according to the mass ratio, stir at a low speed of 60 rpm at 50 °C for 30 min to form a colloidal matrix, slowly add 12 portions of soybean peptide to the colloidal matrix, sprinkle it evenly in 3 times at intervals of 5 min each, maintain the temperature at 50 °C, increase the stirring speed to 80 rpm, and continuously mix for 40 min to ensure that the low molecular weight peptides fully fill the pores of the colloid; Add 12 portions of oyster peptide and 6 portions of wolfberry peptide to 100 portions of water, a premixed solution stirred at 200 rpm for 10 min, add it in two times at intervals of 10 min each, and finally add L-arginine to adjust the PH to 6.8, stir at 50 °C and 60 rpm at a low speed for 2 h to form a homogeneous peptide micelle; Add 7 portions of cordyceps militaris peptide and 15 portions of L-arginine to 50 portions of water, control the liquid temperature at 35 °C, use a high-speed shear emulsifier, disperse at 3000 rpm for 15 min until completely dissolved to obtain a mixed solution of cordyceps militaris peptide and L-arginine;

[0048] (4) Compound and drying: Preheat the epimedium-morinda officinalis concentrate to 40 °C, turn on the magnetic stirrer at 300 rpm, stir for 5 min and then add the ginseng-Eucommia ulmoides male flower concentrate, raise the temperature to 45 °C, stir at a constant temperature for 10 min, then cool down to 35 °C, add the above homogeneous peptide micelle at a flow rate of 10 mL / min, switch to a homogenizer at 8000 rpm for 3 min, then add the above mixed solution of cordyceps militaris peptide and L-arginine at a flow rate of 5 mL / min, mix at a low speed of 200 rpm at room temperature for 20 min, then add 8 portions of maca extract, raise the temperature to 35 °C, stir at 400 rpm for 15 min, and finally dry in an 80 °C environment for 30 min.

[0049] Example 3

[0050] (1) Ethanol fractional extraction: First - stage extraction: 18 parts of epimedium powder ground to 80 mesh were added with 20 times the mass of 70% ethanol by mass ratio, refluxed at 65 °C for 40 min to obtain filtrate A and residue A; Second - stage extraction: Residue A and 12 parts of morinda officinalis powder ground to 80 mesh were added with 15 times the mass of 50% ethanol by mass ratio, and extracted by ultrasonic wave at 45 °C with a frequency of 40 kHz for 30 min to obtain filtrate B and residue B; Filtrates A and B were combined and concentrated under reduced pressure at a pressure of - 0.08 MPa and a pulse frequency of 0.5 Hz, at a temperature of 60 °C, until the relative density reached 1.25 to obtain the epimedium - morinda officinalis concentrated solution;

[0051] (2) Dynamic counter - current extraction: Prepare 18 parts of ginseng sliced into 2 - mm - thick slices, add 0.05% pectinase and 50 parts of water, pre - treat and soak at 30 °C for 20 min, then add 25 times the mass of 60% ethanol solution according to the mass ratio with 8 parts of eucommia ulmoides male flower powder ground to 60 mesh, and perform dynamic counter - current extraction at 40 °C with a flow rate of 1.2 L / min for 2 h. The filtrate was rotary - evaporated at 50 °C to recover ethanol to obtain the ginseng - eucommia ulmoides male flower concentrated solution;

[0052] (3) Polypeptide mixing: Take 10 parts of polygonatum sibiricum peptide and 5 parts of pueraria lobata peptide and add them to the reaction kettle, add 80 parts of water according to the mass ratio, stir at a low speed of 60 rpm at 50 °C for 30 min to form a colloidal matrix. Slowly add 12 parts of soybean peptide substance to the colloidal matrix, sprinkle it evenly in 3 times, with an interval of 5 min each time, maintain the temperature at 50 °C, and increase the stirring speed to 80 rpm, and continue to mix for 40 min to ensure that low - molecular - weight peptides fully fill the pores of the colloid; Add 10 parts of oyster peptide and 6 - 8 parts of wolfberry peptide to 100 parts of water, a premixed solution stirred at 200 rpm for 10 min, add it in two times, with an interval of 10 min each time, and finally add L - arginine to adjust the pH to 6.8, stir at 50 °C and 60 rpm at a low speed for 2 h to form a homogeneous peptide micelle; Add 5 parts of cordyceps militaris peptide and 10 parts of L - arginine to 50 parts of water, control the liquid temperature at 35 °C, use a high - speed shear emulsifier, disperse at 3000 rpm for 15 min until completely dissolved to obtain a mixed solution of cordyceps militaris peptide and L - arginine;

[0053] (4) Compound and drying: Preheat the epimedium-morinda officinalis concentrated liquid to 35°C, turn on the magnetic stirrer at 300 rpm, after stirring for 5 min, add the ginseng-epimedium pubescens concentrated liquid, heat up to 45°C, keep stirring at a constant temperature for 10 min, then cool down to 35°C, add the above-mentioned homogenized peptide micelles at a flow rate of 10 mL / min, switch to a homogenizer at 8000 rpm for 3 min, then add the above-mentioned mixed solution of cordyceps militaris peptide and L-arginine at a flow rate of 5 mL / min, mix at a low speed of 200 rpm at room temperature for 20 min, then add 10 parts of maca extract, heat up to 35°C, stir at 400 rpm for 15 min, and finally dry in an 80°C environment for 30 min.

[0054] Comparative Example 1

[0055] The difference between this comparative example and Example 1 is that in this comparative example, fractional ethanol extraction is not used. The specific ethanol fractional extraction is as follows: Add 20 times the mass fraction of 70% ethanol to 19 parts of epimedium powder crushed to 80 meshes and 13 parts of morinda officinalis powder crushed to 80 meshes according to the mass ratio, reflux at 62°C for 40 min to obtain a filtrate and a filter residue; Concentrate the filtrate under reduced pressure at a pressure of -0.08 MPa and a pulse frequency of 0.5 Hz, at a temperature of 60°C, to a relative density of 1.25 to obtain the epimedium-morinda officinalis concentrated liquid; The rest is the same as Example 1.

[0056] Comparative Example 2

[0057] The difference between this comparative example and Example 1 is that in this comparative example, ginseng and epimedium pubescens are directly extracted and concentrated with an ethanol solution. Specifically: Prepare 16 parts of ginseng sliced into 2 mm thick slices, add 0.05% pectinase, 50 parts of water, pretreat and soak at 30°C for 20 min, then add 25 times the mass fraction of 60% ethanol solution to it and 10 parts of epimedium pubescens powder crushed to 60 meshes according to the mass ratio, extract at 40°C and 300 rpm for 2 h, and recover ethanol by rotary evaporation of the filtrate at 50°C to obtain the ginseng-epimedium pubescens concentrated liquid; The rest is the same as Example 1.

[0058] Comparative Example 3

[0059] The difference between this comparative example and Example 1 is that in this comparative example, ginseng is not pretreated with pectinase, etc., and direct dynamic countercurrent extraction is carried out. Specifically: Prepare 16 parts of ginseng sliced into 2 mm thick slices, add 25 times the mass fraction of 60% ethanol solution to it and 10 parts of epimedium pubescens powder crushed to 60 meshes according to the mass ratio, carry out dynamic countercurrent extraction at 40°C, with a flow rate of 1.2 L / min for 2 h, and recover ethanol by rotary evaporation of the filtrate at 50°C to obtain the ginseng-epimedium pubescens concentrated liquid; The rest is the same as Example 1.

[0060] Comparative Example 4

[0061] The difference between this comparative example and Example 1 lies in that the polypeptide mixing in this comparative example is a conventional mixing process: Polypeptide mixing: Take 9 parts of polygonatum sibiricum peptide, 5.5 parts of kudzu root peptide, and 12 parts of soybean peptide. Add 11 parts of oyster peptide and 7 parts of wolfberry peptide to 180 parts of water, stir at 200 rpm for 2 h, and finally add L-arginine to adjust the pH to 6.8. Stir at 60 rpm at 50 °C for 2 h to form a homogeneous peptide micelle; the rest is the same as in Example 1.

[0062] Comparative Example 5

[0063] The difference between this comparative example and Example 1 lies in the different polypeptide mixing processes. The polypeptide mixing in this comparative example is as follows: Polypeptide mixing: Take 9 parts of polygonatum sibiricum peptide and 5.5 parts of kudzu root peptide and add them to the reaction kettle. Add 80 parts of water according to the mass ratio. Stir at 60 rpm at 50 °C for 30 min to form a colloidal matrix. Add 12 parts of soybean peptide, maintain the temperature at 50 °C, and increase the stirring speed to 80 rpm and continue to mix for 40 min to ensure that the low molecular weight peptides fully fill the pores of the colloid; then add 11 parts of oyster peptide and 7 parts of wolfberry peptide, and add 100 parts of water. Stir at 200 rpm for 10 min. Finally, add L-arginine to adjust the pH to 6.8. Stir at 60 rpm at 50 °C for 2 h to form a homogeneous peptide micelle; the rest is the same as in Example 1.

[0064] Comparative Example 6

[0065] The difference between this comparative example and Example 1 lies in that the compounding and drying process in this comparative example is as follows: Mix the epimedium-morinda officinalis concentrated ginseng-epimedium pubescens concentrated liquid, homogeneous peptide micelle, cordyceps militaris peptide and L-arginine, 9 parts of maca extract, and stir at 500 rpm at room temperature for 30 min. Finally, place it in an 80 °C environment and dry for 30 min; the rest is the same as in Example 1.

[0066] Comparative Example 7

[0067] The difference between this comparative example and Example 1 lies in that the compounding and drying process in this comparative example is as follows: Preheat the epimedium-morinda officinalis concentrated liquid to 38 °C, turn on the magnetic stirrer at 300 rpm, stir for 5 min and then add the ginseng-epimedium pubescens concentrated liquid. Add the above homogeneous peptide micelle, stir at 400 rpm for 20 min, and then add the mixed solution of the above cordyceps militaris peptide and L-arginine at a flow rate of 5 mL / min. Mix at 200 rpm at room temperature for 20 min, then add 9 parts of maca extract, raise the temperature to 35 °C, stir at 400 rpm for 15 min, and finally place it in an 80 °C environment and dry for 30 min; the rest is the same as in Example 1.

[0068] Experiment 1:

[0069] Experimental grouping: After male SD rats were fed with normal feed for 14 days for adaptation, blood was collected from the fundus venous plexus of the rats' eyes using a glass capillary tube. The supernatant was obtained by centrifugation. According to the serum uric acid values of the experimental animals, they were randomly divided into 13 groups, namely the normal control group, the model control group, the positive drug (Jinkui Shenqi Pills) group, the groups of Examples 1 - 3, and the groups of Comparative Examples 1 - 7, with 10 rats in each group. There was no significant difference in the basic serum uric acid values among the groups (P > 0.05). Except for the normal group which was given normal feed, the other groups were given feed containing yeast extract and adenine every day, and they were allowed to freely eat and drink during the experiment.

[0070] Pathological model: Except for the normal group which was given normal feed, the other groups were given feed containing yeast extract and adenine every day. Preparation of the high - purine feed: Yeast extract and adenine were evenly mixed into the ground rat pellet feed at contents of 10% and 0.1% respectively, and then re - pelleted to obtain the high - purine feed.

[0071] Administration method: At the same time of modeling, each group was given the corresponding drug by gavage. The dosage was 1.5 g of crude drug / kg body weight / d. The control group was given an equal volume of normal saline. Before administration, it was dissolved in normal saline at a dosage of 30 mg / kg / d and gavaged once a day for 4 consecutive weeks. The normal control group and the model control group were given the corresponding volume of distilled water.

[0072] Detection indexes: Every 2 weeks, the rats were fasted but allowed to drink water for 12 h. Blood was collected from the fundus venous plexus of the rats' eyes using a glass capillary tube, and the serum was separated. The contents of uric acid (UA), blood urea nitrogen (BUN), and creatinine (CR) in the serum were detected by an automatic biochemical analyzer, and the activity of serum XOD was detected using a kit. The effects on the levels of serum uric acid and related indexes in rats after 2 weeks of drug administration are shown in Table 1, and the effects on the levels of serum uric acid and related indexes in rats after 4 weeks of drug administration are shown in Table 2.

[0073] Table 1

[0074] Group Serum UA (mg / kg) Serum BUN (mmol / l) Serum CR (μmol / l) Normal control group 87.04±9.85 6.67±0.68 47.89±8.47 Model control group 99.82±15.47 6.48±0.67 46.18±6.92 Positive drug group 93.58±9.47 6.60±0.45 45.85±8.74 Example 1 87.54±11.50 6.21±0.52 45.12±4.18 Example 2 87.84±12.08 6.23±0.50 45.29±4.35 Example 3 88.13±11.59 6.28±0.58 45.38±4.52 Comparative example 1 92.47±14.87 6.54±0.61 46.45±5.24 Comparative example 2 92.06±16.59 6.41±0.62 46.85±5.37 Comparative example 3 91.84±13.82 6.46±0.63 46.43±4.89 Comparative example 4 91.47±16.79 6.42±0.65 46.81±4.68 Comparative example 5 89.52±12.41 6.32±0.66 45.97±4.52 Comparative example 6 90.86±16.98 6.49±0.62 46.56±4.38 Comparative example 7 89.97±14.06 6.35±0.65 46.26±4.43

[0075] Note: Compared with the normal control group, △ P < 0.05, △△ P < 0.01; compared with the model control group, * P < 0.05, ** P < 0.01

[0076] Table 2

[0077] Group Serum UA (mg / kg) Serum BUN (mmol / l) Serum CR (μmol / l) Serum XOD (μ / l) Normal control group 72.85±5.54 6.84±0.74 57.74±2.84 31.53±3.14 Model control group <![CDATA[88.47±5.58 △△ > 7.48±0.81 58.69±5.74 <![CDATA[38.42±4.25 △△ > Positive drug group <![CDATA[78.21±5.67 ** > <![CDATA[6.72±0.61 * > 56.42±3.25 <![CDATA[35.74±2.48 * > Example 1 <![CDATA[73.54±8.74 * > <![CDATA[6.05±0.54 * > <![CDATA[51.46±3.69 * > <![CDATA[32.58±2.56 * > Example 2 <![CDATA[73.56±7.94 * > <![CDATA[6.12±0.59 * > <![CDATA[51.53±3.58 * > <![CDATA[32.86±2.64 * > Example 3 <![CDATA[73.91±8.51 * > <![CDATA[6.17±0.58 * > <![CDATA[52.47±3.89 * > <![CDATA[33.17±3.15 * > Comparative example 1 <![CDATA[77.51±8.64 * > <![CDATA[6.65±0.68 * > <![CDATA[54.86±4.21 * > <![CDATA[35.51±3.28 * > Comparative example 2 <![CDATA[78.95±9.74 * > <![CDATA[6.49±0.65 * > 55.28±4.58 <![CDATA[34.87±3.35 * > Comparative example 3 <![CDATA[76.51±8.79 * > <![CDATA[6.59±0.72 * > 56.10±4.67 <![CDATA[35.09±3.61 * > Comparative example 4 <![CDATA[76.18±7.49 * > <![CDATA[6.70±0.75 * > <![CDATA[54.68±4.85 * > <![CDATA[34.72±3.58 * > Comparative example 5 <![CDATA[74.28±7.58 * > <![CDATA[6.38±0.61 * > <![CDATA[52.17±4.16 * > <![CDATA[33.95±2.86 * > Comparative example 6 <![CDATA[76.53±8.18 * > <![CDATA[6.56±0.69 * > <![CDATA[55.24±4.92 * > <![CDATA[35.18±3.38 * > Comparative example 7 <![CDATA[74.67±8.56 * > <![CDATA[6.41±0.68 * > <![CDATA[52.34±4.23 * > <![CDATA[34.03±2.69 * >

[0078] Note: Compared with the normal control group, △ P < 0.05, △△ P < 0.01; compared with the model control group, * P < 0.05, **P < 0.01, Experiment 2:

[0079] Experimental grouping: 120 male SD rats, weighing 220 - 240 g, were randomly divided into a control group, a positive drug group, an example group, and a comparative example group, with 10 rats in each group.

[0080] Test drugs: Prepared from Examples 1 - 3 and Comparative Examples 1 - 7, the positive drug group was given Jinkui Shenqi Pills.

[0081] Dose design: For rats in Examples 1 - 3, Comparative Examples 1 - 7, and the positive drug group, the drug dosage was 1.5 g of crude drug / kg body weight / d. The control group was given an equal volume of normal saline. Before administration, it was dissolved in normal saline at a dosage of 30 mg / kg / d and administered by gavage once a day for 30 consecutive days. Rats in each group were fed with ordinary feed and allowed to eat and drink freely.

[0082] Method for inducing estrus in female rats: 48 h before the experiment, female rats were subcutaneously injected with 10 μg of estradiol (dissolved in 0.1 ml of safflower oil); 2 h before the experiment, 10 μg of progesterone was subcutaneously injected. When female rats were in estrus, they showed lordosis and ear flapping when contacting male rats.

[0083] Electrophysiological determination of sexual activity: Rats were fixed in the dorsal position, the biological function experimental system program was set to the electrical stimulation mode, the stimulating electrode was placed on the penis and near the anus of the lower abdomen of the rats, electrical stimulation was given, and the erection latency and ejaculation latency of the rats were measured (the time required from the start of stimulation to penile erection was the erection latency; the time from penile erection to the start of ejaculation was the ejaculation latency).

[0084] Mating test: Male rats and estrous female rats were caged together, one pair per cage, and the sexual activity of male rats was observed from 7 - 9 pm at night. One male rat was placed in the cage for 10 min to adapt, and one estrous female rat was put in, and the time was immediately recorded and the sexual behavior of the male rat was observed within 30 min. The following indicators were recorded: ① The time from the female rat being put in to the male rat's first capture of the female rat (mounting latency). ② The number of mounting actions of the male rat within 30 min (mounting frequency). ③ The number of times the male rat licked the penis after successful mounting within 30 min (ejaculation frequency). ④ The time from the male rat's successful mounting to licking the penis (ejaculation latency). See Tables 3 and 4 for details.

[0085] Table 3

[0086]

[0087]

[0088] Note: Compared with the control group, ＊ p < 0.05; compared with the positive drug group, # p < 0.05.

[0089] Table 4

[0090] Group Erection latency (s) Ejaculation latency (s) Control group 118.59±15.47 250.64±30.59 Positive drug group <![CDATA[79.62±20.68 * > <![CDATA[331.26±27.45 * > Example 1 <![CDATA[45.28±26.84 ＊# > <![CDATA[384.25±16.58 ＊# > Example 2 <![CDATA[46.93±27.41 *# > <![CDATA[383.62±16.83 ＊# > Example 3 <![CDATA[46.85±25.92 *# > <![CDATA[385.93±19.67 ＊# > Comparative example 1 <![CDATA[74.62±22.56 ＊# > <![CDATA[346.82±17.92 *# > Comparative example 2 <![CDATA[78.29±23.58 ＊# > <![CDATA[351.24±16.84 ＊# <!-- 9 -->]]> Comparative example 3 <![CDATA[65.34±24.69 *# > <![CDATA[358.92±19.67 *# > Comparative example 4 <![CDATA[67.25±21.27 ＊# > <![CDATA[362.61±18.43 *# > Comparative example 5 <![CDATA[53.67±23.61 ＊# > <![CDATA[374.53±17.86 *# > Comparative example 6 <![CDATA[57.92±25.82 *# > <![CDATA[368.58±18.29 *# > Comparative example 7 <![CDATA[54.81±24.15 ＊# > <![CDATA[371.69±17.81 *# >

[0091] Note: Compared with the control group, ＊ p < 0.05; compared with the positive drug group, # p < 0.05.

Claims

1. A Chinese medicine composition for nourishing and regulating kidney function, characterized in that: The process is: Ethanol graded extraction: primary extraction: crush 18-20 parts of epimedium to 80 mesh, add 20 times of ethanol according to the mass ratio, reflux at 60-65°C for 40 minutes, and obtain filtrate A and residue A; secondary extraction: crush residue A and 12-15 parts of Morinda officinalis to 80 mesh, add 15 times of 50% ethanol according to the mass ratio, extract at 45°C with ultrasound at 40kHz for 30 minutes to obtain filtrate B and residue B; combine filtrates A and B, and perform reduced pressure concentration at a pressure of -0.08MPa and a pulse frequency of 0.5Hz at a temperature of 60°C, and concentrate to a relative density of 1.25 to obtain an epimedium-Morinda officinalis concentrate; Add 5-7 parts of Cordyceps militaris peptide and 10-15 parts of L-arginine to 50 parts of water, control the liquid temperature to 35°C, use a high-speed shear emulsifier, disperse at 3000 rpm for 15 minutes until completely dissolved to obtain a mixed solution of Cordyceps militaris peptide and L-arginine; Compounding and drying: Preheat the epimedium-morinda officinalis concentrate to 35-40°C, turn on the magnetic stirring at 300 rpm, stir for 5 minutes, then add the ginseng-eucommia male flower concentrate, heat to 45°C, stir at constant temperature for 10 minutes, then cool to 35°C, add the homogenized peptide micelles at a flow rate of 10 mL / min, switch to the homogenizer at 8000 rpm for 3 minutes, then add the mixed solution of the above-mentioned Cordyceps militaris peptide and L-arginine at a flow rate of 5 mL / min, mix at a low speed of 200 rpm at room temperature for 20 minutes, then add 8-10 parts of maca extract, heat to 35°C, stir at 400 rpm for 15 minutes, and finally dry at 80°C for 30 minutes.

2. A nourishing and renal function regulating Chinese medicine composition as claimed in claim 1, characterized in that: The mass fraction of ethanol used in the first extraction of the ethanol graded extraction is 70%, and the mass fraction of ethanol used in the second extraction is 50%.

3. A nourishing and renal function regulating Chinese medicine composition as claimed in claim 1, characterized in that: The ginseng-eucommia male flower concentrate comprises a dynamic countercurrent extraction: 15-18 parts of flaky ginseng and 8-12 parts of eucommia male flowers crushed to 60 mesh are added with 25 times the mass fraction of 60% ethanol solution according to a mass ratio, and dynamic countercurrent extraction is performed at 40° C. with a flow rate of 1.2 L / min for 2 hours, and the filtrate is rotary evaporated at 50° C. to recover ethanol, so as to obtain the ginseng-eucommia male flower concentrate.

4. A nourishing and renal function regulating Chinese medicine composition as claimed in claim 3, characterized in that: The ginseng flakes also include the following treatment: preparing ginseng, cutting it into ginseng flakes with a thickness of 2 mm, adding 0.05% pectinase and 50 parts of water, and pre-treating and soaking it at 30° C. for 20 minutes.

5. A nourishing and renal function regulating Chinese medicine composition as claimed in claim 1, characterized in that: The homogeneous peptide micelle comprises: adding 8-10 parts of Polygonatum sibiricum peptide and 5-6 parts of Pueraria root peptide into a reactor, adding 80 parts of water according to the mass ratio, stirring at a low speed of 60 rpm for 30 minutes at 50°C to form a colloidal matrix, slowly adding 12 parts of soybean peptide into the colloidal matrix, evenly sprinkling it in 3 times, each time with an interval of 5 minutes, maintaining the temperature at 50°C, increasing the stirring speed to 80 rpm, and continuously mixing for 40 minutes to ensure that the low molecular weight peptides fully fill the colloidal pores; adding 10-12 parts of oyster peptide and 6-8 parts of wolfberry peptide to 100 parts of water, stirring the premix at 200 rpm for 10 minutes, adding twice, each time with an interval of 10 minutes, and finally adding L-arginine to adjust the pH to 6.8, stirring at a low speed of 50°C and 60 rpm for 2 hours to form homogeneous peptide micelles.

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