CN116139188A - Preparation method and quality detection method of traditional Chinese medicine composition with effects of tonifying kidney, strengthening spleen, nourishing blood and tranquillizing - Google Patents
Preparation method and quality detection method of traditional Chinese medicine composition with effects of tonifying kidney, strengthening spleen, nourishing blood and tranquillizing Download PDFInfo
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- CN116139188A CN116139188A CN202211720108.9A CN202211720108A CN116139188A CN 116139188 A CN116139188 A CN 116139188A CN 202211720108 A CN202211720108 A CN 202211720108A CN 116139188 A CN116139188 A CN 116139188A
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Abstract
A preparation method and quality detection method of a traditional Chinese medicine composition with the effects of tonifying kidney and strengthening spleen, nourishing blood and tranquillizing, which relate to the field of traditional Chinese medicines, are prepared by the processes of weighing, extracting, concentrating, mixing, granulating, tabletting and the like. Simple process operation, short production period and lower cost, and is suitable for industrialized mass production. The invention establishes a method for detecting the extract content of the product, a method for detecting the content of effective substances and a method for identifying the effective substances, and the method for detecting the extract content can effectively control the quality of the pharmaceutical preparation. The identification method eliminates the interference substances, can rapidly and effectively identify the products, and has simple method and strong specificity. Compared with the existing products, the product has obvious drug effect, high bioavailability and high clinical effective rate through clinical application comparison data.
Description
Technical Field
The invention belongs to the field of traditional Chinese medicines, and in particular relates to a preparation method and a quality detection method of a traditional Chinese medicine composition with the effects of tonifying kidney, strengthening spleen, nourishing blood and soothing nerves.
Background
The heart governs blood vessels and is a principal organ, which is the most important and mental part of the five zang-organs and six fu-organs. Heart stores spirit, and spirit plays an important role in life activities of human body, and malnutrition of heart spirit is manifested by palpitation, shortness of breath, dizziness, insomnia, amnesia, lusterless complexion, listlessness, debilitation, anorexia, pale red tongue and weak pulse. Kidneys are congenital origin, are root of human growth and development, and function of viscera to nourish viscera and tissues. Spleen is the acquired root and is responsible for transporting and transforming water-dampness and transporting and transforming food essence to the viscera and tissues of the whole body. If kidney qi is deficient or kidney yang is deficient, spleen yang cannot be warmed or yang is deficient and water is flooded, spleen yang is injured, water dampness cannot be transformed, kidney disease and spleen are not transformed, and finally spleen-kidney deficiency or spleen-kidney yang deficiency is caused.
The deficiency of both the spleen and kidney and malnutrition of the heart and mind can cause symptoms such as palpitation, insomnia, inappetence, listlessness, frequent urination, impotence, neurasthenia, non-organic sexual hypofunction, female climacteric syndrome and the like.
Insomnia is accompanied by symptoms of multiple systems and viscera, and is mainly caused by viscera dysfunction, qi and blood deficiency, heart blood deficiency, and heart-mind failing to nourish and calm, so insomnia is mainly caused by heart, and therefore palpitation is caused by most patients. From the etiology and pathogenesis of insomnia, it is considered that the disease is caused by either malnutrition of heart spirit or disturbance of heart spirit by pathogenic factors. Insomnia is the main symptom of neurasthenia. Neurasthenia is caused by unbalance of the excitation and inhibition processes of cerebral cortex, and relates to symptoms such as 'depression', 'palpitation', 'insomnia', 'deficiency loss', 'impotence' and the like in traditional Chinese medicine, which are caused by deficiency of heart and spleen, hyperactivity of fire due to yin deficiency, timidity due to deficiency of heart and incoordination in stomach. It is mainly characterized by easy fatigue, easy excitation, mood disorder, sleep disorder, tension pain (common head and muscle tension pain, stiff neck, waist soreness and backache; autonomic nerve disorder can be seen as chest distress, short breath, palpitation, hyperhidrosis, cold limbs, anorexia, frequent urination, spermatorrhea, impotence, premature ejaculation, irregular menstruation, etc.), anxiety, and suspected heart disease. Therefore, replenishing qi to invigorate the spleen, purging excess and tonifying deficiency, nourishing blood and tranquillizing is a key for treating palpitation and insomnia and neurasthenia caused by deficiency of both spleen and kidney and malnutrition of heart spirit.
Inappetence, food stagnation, dyspepsia and infantile anorexia are generally caused by food stagnation damaging spleen, food stagnation and gastrointestinal or spleen qi deficiency. Qi is not epidemic due to spleen deficiency, and qi is stagnant and accumulated due to qi failure. If the accumulation is due to spleen deficiency, the patients can not strengthen the efficacy of the medicine, or conduct the concurrent tonic action, or supplement more and less, or supplement first and then remove. Thus, regulating anorexia is actually regulating spleen and stomach function.
Climacteric syndrome is a common disorder in women, and is well developed in menopausal women. Female ovary hypofunction, thereby leading to estrogen level reduction, and adverse reactions such as metabolic dysfunction, nerve dysfunction and the like can be caused when patients are ill. The traditional Chinese medicine considers that the pathological basis of the climacteric syndrome is viscera dysfunction such as liver and kidney, and the viscera dysfunction is related to heart failure and nourishment, heart and liver fire hyperactivity, kidney essence deficiency, spleen qi deficiency and other factors. The disease is in the pathological state of imbalance of yin and yang of the five zang organs, kidney essence deficiency, essence qi deficiency, ren Chong imbalance, pulse and conception vessel qi and blood deficiency. Kidney yin deficiency is the main clinical manifestation and is mainly characterized by autonomic dysfunction, and spleen is the root cause of morbidity and acquired root cause respectively. The symptoms of weakness of spleen and stomach are common as blood and qi biochemical sources, spleen and yang deficiency, stomach qi deficiency and other spleen and stomach weakness symptoms directly cause deficiency of qi and blood, so that kidney essence is deficient, essence and blood are weakened mutually, qi and blood are insufficient, and the weather is failure, so that a plurality of lesions of climacteric women appear, such as menstrual disorder, dysphoria, sweating, hectic fever, flushed face, poor sleep, arthralgia and the like, and serious patients can also cause cardiovascular and cerebrovascular diseases, atrophic urethritis, vaginitis, osteoporosis, fracture and the like.
The symptoms are treated by western medicine sedative drugs and hormone drugs, and diseases such as obesity, cardiovascular diseases, apoplexy, endometrial cancer and the like can be induced after long-term use. It is therefore of great clinical importance to find an effective and safe method. The traditional Chinese medicine treatment methods are various, including treatment based on syndrome differentiation, acupuncture, physiotherapy, psychological dispersion and the like, and have remarkable effects and low adverse reaction rate. The traditional Chinese medicine can be clinically combined with western medicines for treatment, and has obvious effect of improving symptoms and signs.
Disclosure of Invention
The invention aims to provide a preparation method and a quality detection method of a traditional Chinese medicine composition with the effects of tonifying kidney, strengthening spleen, nourishing blood and tranquillizing. The preparation method provided can effectively improve the content of the effective substances of the medicine.
The invention is prepared from acanthopanax extract, schisandra extract, pilose antler and other Chinese medicinal preparations, has the effects of tonifying kidney and spleen, nourishing blood and tranquillizing, and is used for treating palpitation and insomnia, inappetence, listlessness, frequent urination and impotence, neurasthenia, non-organic sexual dysfunction and female climacteric syndrome caused by malnutrition of spleen and kidney.
The preparation method of the traditional Chinese medicine composition with the effects of tonifying kidney, strengthening spleen, nourishing blood and soothing nerves comprises the following steps:
Step one, extraction of acanthopanax extract
Pulverizing radix Acanthopanacis Senticosi into coarse powder, adding 6-8 times of 75% ethanol, reflux extracting for 1-3 hr, and filtering to obtain filtrate 1; extracting the residues for 20-40 min under the assistance of ultrasonic waves at 50-70 ℃ with ultrasonic power of 300W, and filtering to obtain filtrate 2; dissolving the residue in 10 times of water, extracting with microwave for 20-40 min under the assistance of microwave power of 450w, and filtering to obtain filtrate 3; mixing filtrate 1, filtrate 2 and filtrate 3, concentrating, drying, and weighing;
step two, extraction of schisandra chinensis fluid extract
1) Crushing: taking weighted shizandra berry, and putting the shizandra berry into a crusher for crushing;
2) Wetting: placing the weighed coarse schisandra chinensis powder into a clean container, wherein the mass volume ratio is 1kg: 600-700 mL of ethanol with the proportion volume percentage of 40% is uniformly stirred and placed in a closed state for 2 hours;
3) Dipping: placing a filtering device at the bottom of the percolation tank, closing a discharging door, filling the wet schisandra chinensis medicinal material into the percolation tank, covering a layer of 100-mesh silk cloth on the material surface, and firmly pressing the silk cloth by using a stainless steel pore plate; opening a liquid discharge valve below the percolation tank, adding ethanol solution with the volume percentage of 40% from the top of the percolation tank, and discharging air in a decoction piece gap; adding 40% ethanol solution until the liquid medicine flows out from the bottom and the solvent is 2-3cm higher than the surface of the medicinal materials, closing the liquid discharge valve, placing the flowing liquid medicine into the percolating tank, covering the percolating tank with an upper cover, and hermetically soaking for 24h;
4) Percolating: opening a liquid inlet valve and a liquid discharge valve, starting to percolate at the speed of 2-3 mL/kg of medicinal materials per minute, adding ethanol solution with the volume percentage of 40% and the volume of 8 times of the percolate material into the percolate, collecting the primary percolate with the decoction piece quantity of 85%, and finally collecting the continuous percolate until the color is light and nearly colorless for later use;
5) Concentrating: concentrating the percolate of the step 4) to extract of 1.35-1.40;
6) Preparing: cooling the shizandra extract to room temperature, mixing with the primary percolate, stirring uniformly, adding 40% ethanol by volume percent, and adjusting to decoction pieces: fluid extract=1:1, standing for 24h, and collecting supernatant;
step three, extracting the cartialgenous extract with the mass percentage of 10 percent
7) Adding ethanol solution with volume percentage of 50% to 2-10 times of the feeding amount of cornu Cervi Pantotrichum, soaking for 5-24 hr, timing when the liquid medicine boils, and reflux extracting for 2-6 hr;
8) Taking out the cornu Cervi Pantotrichum after reflux extraction, and removing skin while hot; adding peeled cornu Cervi Pantotrichum into extraction tank, adding ethanol solution with volume percentage of 50% 2-10 times of the crude drug, and reflux extracting for 2-6 hr;
9) Repeating step 8) for 2-6 times, collecting supernatant each time, mixing the supernatants after reflux extraction for 2-6 times, recovering ethanol, concentrating cornu Cervi Pantotrichum extractive solution to obtain medicinal liquid with volume ratio of 1:1;
Adding 5-12 times of 95% ethanol solution by volume percentage into the concentrated solution under stirring, standing for 24 hr after the concentrated solution and 95% ethanol solution are stirred uniformly, collecting supernatant, filtering, concentrating to one tenth of crude drug, and packaging into clean container; diluting with water to obtain 60% crude drug solution, filtering, and freezing; thawing at room temperature, filtering, diluting to obtain solution containing crude drug 10% and alcohol 30%, and filtering;
step four, granulating
10 Pre-treatment, weighing and batching
Crushing sodium glycerophosphate, sieving with a 100-mesh sieve, sieving calcium carbonate with a 100-mesh vibrating sieve, sieving starch with a 120-mesh sieve, and sieving magnesium stearate with a 100-mesh sieve;
11 Mixing, granulating, and drying
Mixing the weighed raw materials with the acanthopanax extract extracted in the first step and the schisandra chinensis fluid extract extracted in the second step, adding 1/3 of the total ethanol with the volume percentage of 95%, stirring for 20 minutes to uniformly mix the materials, spraying and mixing the velvet extract with the mass percentage of 10% extracted in the third step by adopting a one-step granulator, continuously adding 1/3 of the total ethanol with the volume percentage of 95% by adopting a spraying mode, stirring for 10 minutes, then adding the ethanol with the residual volume percentage of 95%, stirring for 30 minutes to prepare a soft material, granulating with 20 meshes, drying, sieving with a 18-mesh sieve, adding magnesium stearate, mixing for 30 minutes, tabletting, and coating a film.
Further, the wet shizandra berry medicinal material in the step two 3) is filled into a percolation tank, and the material filling amount is not more than 2/3 of the height of the percolation tank body.
Further, the temperature of the extract in the second step 5) is 50 ℃.
Further, the antler in the step 7) is the upper section of the free-range antler in the Heilongjiang province.
Further, in the step 7), the added volume percentage is 5 times of the feeding amount of the ethanol solution with the volume percentage of 50% to the pilose antler, after soaking for 12 hours, the timing is started when the liquid medicine is boiled, and the reflux extraction is carried out for 4 hours.
Further, taking out the pilose antler after reflux extraction in the step 8), and removing the skin while the pilose antler is hot; adding peeled cornu Cervi Pantotrichum into extraction tank, adding ethanol solution with volume percentage of 50% 5 times of the crude drug, and reflux extracting for 4 hr.
Further, repeating step 8) for 4 times, collecting supernatant, concentrating the supernatant cornu Cervi Pantotrichum extractive solution after reflux extraction for 5 times, recovering ethanol, and concentrating cornu Cervi Pantotrichum extractive solution to obtain medicinal liquid with volume ratio of 1:1.
The extract detection method of the traditional Chinese medicine composition with the effects of tonifying kidney, strengthening spleen, nourishing blood and soothing nerves is carried out according to the following contents:
Taking a traditional Chinese medicine composition tablet, grinding, precisely weighing, and placing into a conical flask according to the volume-mass ratio of 2g: adding 60-80% methanol solution in a volume percentage of 50mL, sealing, weighing, standing for 1h, connecting a reflux condenser, heating to boiling, keeping for 1h, cooling, taking down a conical flask, sealing, weighing again, supplementing the reduced weight with methanol, shaking, filtering with a drying filter, precisely measuring 25mL of filtrate, placing into a evaporating dish dried to constant weight, evaporating in a water bath, drying at 105 ℃ for 3h, cooling in a dryer for 30min, precisely weighing; and calculating the content of the alcohol soluble extract in the sample by using the dried product, thus completing the quality detection method.
The detection method of the effective substances of the traditional Chinese medicine composition with the effects of tonifying kidney, strengthening spleen, nourishing blood and soothing nerves is carried out according to the following contents:
1) Preparation of reference substance solution
Respectively weighing syringin, eleutheroside E, isofraxidin, schizandrin A, schizandrin B, schizandrin A and schizandrin B reference substances, respectively adding methanol for dissolving to obtain reference substance stock solutions with concentration of 1.0mg/mL, precisely sucking the reference substance stock solutions, and adding methanol to obtain mixed solution with concentration of 50 μg/mL; respectively and precisely absorbing the control stock solutions of syringin, eleutheroside E, isofraxidin, schizandrin A, schizandrin B, schizandrin A and schizandrin B, and adding methanol to prepare control solutions with the concentration of 0.2mg/mL, 0.1mg/mL, 0.3mg/mL, 0.1mg/mL and 0.2 mg/mL;
2) Preparation of test sample solution
Taking a traditional Chinese medicine composition tablet with the effects of tonifying kidney and strengthening spleen, nourishing blood and tranquillizing, grinding, precisely weighing 2g, precisely weighing, placing in a conical bottle with a plug, precisely adding 50mL of 60% -80% methanol, sealing, weighing the mass, performing ultrasonic treatment for 10min to dissolve, cooling, weighing the mass, supplementing the lost mass with methanol, shaking uniformly, and filtering with a 0.45 microporous filter membrane to obtain a sample solution;
an Agilent ZB-Extend C18 chromatographic column is adopted, 250mmx4.6mm,5 mu m; mobile phase A is 0.8% phosphoric acid water solution, mobile phase B is acetonitrile, and the flow rate is 0.8mL/min; the detection wavelength is 220nm; the sample injection amount is 20uL; the column temperature is 40 ℃, and gradient elution is carried out according to the following mode, namely the detection of the effective substances is completed;
the identification method of the traditional Chinese medicine composition with the effects of tonifying kidney, strengthening spleen, nourishing blood and soothing nerves is carried out according to the following content:
grinding the Chinese medicinal composition tablet, and placing in a test tube with a stopper; the volume mass ratio is 1g: adding chloroform into 15mL, performing ultrasonic treatment for 20min, filtering, evaporating filtrate to dryness, and adding chloroform into residue 1mL to dissolve to obtain sample solution; preparing 1g of acanthopanax reference medicine, and preparing a reference medicine solution by the same method; according to thin layer chromatography test, 50ul of each of the above two solutions is absorbed and respectively spotted on the same silica gel G thin layer plate, cyclohexane chloroform-ethyl acetate-methanol is used as developing agent, and then developed, taken out, dried, inspected under 365nm ultraviolet lamp, and fluorescent spots with the same color are displayed on the positions corresponding to the reference substance chromatogram in the sample chromatogram.
Further, the volume ratio of petroleum ether-n-hexane-ethyl acetate-methanol is 8:10:3:2.
Compared with the prior art, the invention has the beneficial effects that:
1. the traditional Chinese medicine composition with the effects of tonifying kidney, strengthening spleen, nourishing blood and soothing nerves is mainly prepared by the processes of weighing, extracting, concentrating, mixing, granulating, tabletting and the like. Simple process operation, short production period and lower cost, and is suitable for industrialized mass production.
2. The invention provides a prescription of a traditional Chinese medicine composition with the effects of tonifying kidney, strengthening spleen, nourishing blood and tranquillizing. Comprises radix Acanthopanacis Senticosi extract, fructus Schisandrae fluid extract, 10% cornu Cervi Pantotrichum extract, and sodium glycerophosphate. In the invention, the effective substances in the acanthopanax extract, the schisandra chinensis fluid extract and 10% pantocrin are improved by effective means to play a synergistic effect, so that the extract content in the medicinal preparation is improved by 1.8 times, and the treatment effect of the medicinal preparation is greatly improved.
3. Compared with the traditional extraction method, the invention adopts reflux and ultrasonic wave to extract the acanthopanax, the total flavone content of the acanthopanax is increased by more than 3 times, the DPPH free radical clearance is obviously improved, the antioxidation activity is better, the extraction efficiency of the acanthopanax extract is greatly improved, and the extraction time is shortened. Provides a technical foundation for the process mass production.
4. The invention improves the extraction process of the schisandra chinensis fluid extract, adopts the methods of crushing, wetting, soaking and the like to jointly extract, and greatly improves the effective medicinal components in the schisandra chinensis fluid extract.
5. The invention improves the extraction process of the pilose antler, selects the upper section of pilose antler of Jilin province (free-ranging) as the raw material, and greatly improves the content of active substances in the pilose antler extract by the operations of medicinal material soaking, extraction with skin, peeling extraction, standing leaching with 95% ethanol and the like, thereby improving the efficacy of the traditional Chinese medicine composition.
6. According to the invention, a one-step granulator is adopted for spray mixing, and 95% ethanol is added for multiple times in a spray mode, so that the difficulty that the extract is large in quantity and not easy to mix uniformly in the granulating process is effectively solved, the average content of the medicinal preparation prepared by the invention in the extract content detection result is 35.2%, the RSD is 2.4%, and compared with the traditional process, the RSD is reduced by 18%, and the effective substances in the medicinal preparation are evenly distributed, so that the treatment effect is stable.
7. The invention establishes a method for detecting the extract content of the product, a method for detecting the content of effective substances and a method for identifying the effective substances, and the method for detecting the extract content can effectively control the quality of the pharmaceutical preparation; the method for detecting the content of the effective substances can detect the effective components in the acanthopanax and the schisandra chinensis at the same time, and the contents of syringin, acanthopanaxoside E, isofraxidin, schizandrin A, schizandrin B, schizandrin A and schizandrin B are effectively separated and detected in a gradient elution mode, so that the effective components in the pharmaceutical preparation can be quantitatively detected; the identification method eliminates the interference substances, can rapidly and effectively identify the products, and has simple method and strong specificity.
8. Compared with the existing products, the product has obvious drug effect, high bioavailability and high clinical effective rate through clinical application comparison data.
Drawings
FIG. 1 is a graph of a mixed solution; wherein 1 is syringin; 2 is eleutheroside E;3 is isofraxidin; 4 is schisandrin A; 5 is schisandrin B; 6 is schizandrin A; 7 is schizandrin B.
Detailed Description
It will be understood by those of ordinary skill in the art that the foregoing embodiments are specific examples of carrying out the invention and that various changes in form and details may be made therein without departing from the spirit and scope of the invention.
For the purposes of clarity, technical solutions and advantages of embodiments of the present invention, the following detailed description will clearly illustrate the spirit of the present disclosure, and any person skilled in the art, after having knowledge of the embodiments of the present disclosure, may make alterations and modifications to the technology taught by the present disclosure without departing from the spirit and scope of the present disclosure.
The exemplary embodiments of the present invention and the descriptions thereof are intended to illustrate the present invention, but not to limit the present invention.
Example 1
Extraction of acanthopanax extract
The acanthopanax contains more active ingredients, mainly contains phenolic glycoside, triterpenes, polysaccharide, flavone, lignan, coumarin, various trace elements, amino acids and the like. Wherein the total flavone in the acanthopanax is one of main antioxidant components of the acanthopanax, and plays an important role in improving the immunity of organisms, resisting oxidation and the like. The flavonoid compounds in the acanthopanax medicinal material have very complex structures, and mainly comprise quercetin, quercitrin, hyperin, rutin and the like. The ethanol reflux method for extracting flavonoid compounds has the defects of long time consumption, large solvent consumption, multiple extraction times and low extraction rate, and the biological activity of flavonoid compounds is easily reduced after repeated high temperature for a long time. The ultrasonic auxiliary method can overcome the defects of the traditional extraction process through the mechanical, cavitation and thermal effects of ultrasonic waves, and can effectively improve the extraction rate of flavonoid compounds.
The embodiment optimizes the extraction process, adopts reflux and ultrasonic wave to extract the acanthopanax by cooperating with microwave, greatly improves the extraction efficiency of the acanthopanax extract and shortens the extraction time. Meanwhile, the flavonoid active ingredients in the extract are added, and a technical basis is provided for the process mass production.
200g of acanthopanax root is taken as a control group, crushed into coarse powder, added with 7 times of 75% ethanol, extracted for 12 hours under reflux, filtered, and the filtrate is recovered with ethanol, concentrated, dried and weighed for measurement.
200g of acanthopanax root is taken in an experimental group 1, crushed into coarse powder, added with 7 times of 75% ethanol, extracted for 5 hours under reflux, filtered, and the ethanol is recovered from the filtrate, concentrated and dried. Extracting the residue with 10 times of water for 2 times each for 2 hr, concentrating the supernatant, drying, mixing the above dry extracts, and weighing.
In the experimental group 2, 200g of acanthopanax is crushed into coarse powder, 7 times of 75% ethanol is added, reflux extraction is carried out for 2 hours, and filtration is carried out to obtain filtrate 1; extracting the residues with ultrasound for 30min at 60deg.C under ultrasonic power of 300W, and filtering to obtain filtrate 2; dissolving the residue with 10 times of water, extracting with microwave for 30min under the assistance of microwave power of 450w, and filtering to obtain filtrate 3. And combining the filtrate 1, the filtrate 2 and the filtrate 3, concentrating, drying, and weighing and measuring.
The experimental results are shown in table 1:
table 1 results of extraction process experiments
Compared with the traditional reflux extraction process, the experimental result shows that the reflux and ultrasonic wave are adopted to extract the acanthopanax root by combining the microwaves, so that the active ingredients are released to the greatest extent, and the extraction efficiency of the active ingredients such as syringin is further improved. The loss of active ingredients in the original extraction process is reduced, the extraction time is shortened, the dry paste rate is improved, and the biological activity of the product is reserved to a greater extent.
DPPH radical scavenging method:
diluting radix Acanthopanacis Senticosi extract with 80% ethanol to a certain concentration, mixing with 0.3mmol/l DPPH solution, standing at room temperature in dark place for 30min, measuring absorbance at 517nm, and calculating clearance. Absolute ethanol was used as a blank.
Clearance (%) = [ a- (B-C) ]/a×100%
Wherein A: blank absorbance values; b: the light absorption value of the acanthopanax extract group; c: background group absorbance values of acanthopanax extract group.
The acanthopanax extract prepared by the test is detected according to the DPPH free radical clearance measuring method, and the detection result is shown in table 2.
TABLE 2 extraction process experimental results
Compared with the traditional extraction method, the reflux and ultrasonic wave synergistic microwave extraction method is adopted to extract the acanthopanax, the total flavone content is increased by more than 3 times, the DPPH free radical clearance is increased by 3 times, and the oxidation resistance is greatly enhanced, so that the extraction process can furthest maintain the content and the bioactivity of the functional components of the product, and has great advantages compared with the traditional extraction method. The antioxidant activity of the product is improved, and the product has better curative effects on improving the immunity of organisms, resisting aging, relieving visual fatigue and the like.
Example 2
Extraction of schisandra fruit fluid extract
Experiment 1 existing Process
1. Percolating: removing impurities and fruit stalks, mould grains and dust from fructus Schisandrae chinensis, and sieving. Mixing fructus Schisandrae with 40% ethanol solution, wetting, sealing, standing for 24 hr, placing into a percolating cylinder, adding ethanol with 40% concentration, percolating with ethanol as solvent, removing air from the gap of the medicine to the maximum extent, percolating until the percolate is light and colorless, collecting percolate, filtering, and concentrating.
2. Concentrating: concentrating the percolate respectively in vacuum concentrating tank to appropriate amount, and diluting with 40% ethanol to obtain crude drug: fluid extract=1:1, standing (specific gravity 1.05-1.25), filtering after clarification, and filling the fluid extract into a clean container with a cover.
Experiment 2 the extraction technique of the invention
1. Crushing: taking weighted shizandra berry, and putting the shizandra berry into a crusher for crushing.
2. Wetting: the weighed coarse schisandra chinensis powder is placed in a clean container, stirred uniformly by 40% ethanol (according to the coarse powder: 40% ethanol solution=1 kg:660 ml), and placed in a closed state for 2 hours.
3. Dipping: and placing a filtering device at the bottom of the percolation tank, closing a discharging door, and preventing leakage. The shizandra berry medicinal material is filled into a percolation tank, the medicinal powder is uniform and the tightness is consistent, the material filling amount is not more than 2/3 of the height of the percolation tank body, and a layer of 100-mesh silk cloth is covered on the material surface and firmly fixed by stainless steel Kong Banya. Opening a liquid discharge valve below the percolation tank, slowly adding 40% ethanol solution from the top of the percolation tank, and discharging air in the gap of decoction pieces as much as possible. Adding 40% ethanol solution until the liquid medicine flows out, and sealing the liquid discharge valve when the solvent is 2-3cm higher than the surface of the medicinal materials, placing the flowing liquid medicine into the percolating tank, covering the upper cover, and soaking for 24 hr.
4. Percolating: and (3) opening a liquid inlet valve and a liquid discharge valve, starting to slowly percolate at a speed of about 2-3ml per kilogram of medicinal materials per minute, adding 8 times of 40% ethanol solution (calculated by volume) into the mixture, collecting and storing the primary percolate (unit L) with the decoction piece amount of 85% in another container, and finally collecting and storing the secondary percolate until the color of the secondary percolate is light and colorless for later use.
5. Concentrating: concentrating to extract of about 1.35-1.40 (50 ℃), collecting extract of 11.5kg of fructus Schisandrae chinensis with dosage not more than 1.7L, and collecting extract of 34.5kg of fructus Schisandrae chinensis with dosage not more than 5.1L for use.
6. Preparing: cooling the concentrated extract of the continuous filtrate of the schisandra chinensis to room temperature, mixing with the primary percolate, uniformly stirring, adding 40% ethanol, and adjusting to obtain decoction pieces: fluid extract=1:1, left to stand for 24 hours, and the supernatant was taken and filled into a clean capped container.
The schisandra chinensis streams extracted in the experiment 1 and the experiment 2 are detected according to a second content detection method, the detection results are shown in the following table 3, and the mixed solution map is shown in fig. 1.
TABLE 3 detection results of Schisandra chinensis stream content
Conclusion: according to the test results, the content of schizandrin A, schizandrin B, schizandrin A and schizandrin B in the schisandra fluid extract extracted in the experiment 2 is higher than that in the experiment 1, which indicates that the technology of the experiment 2 can more effectively extract important components in the schisandra.
Example 3
10% of pilose antler extract by mass percentage
Experiment 1 existing extraction Process
Cutting cornu Cervi Pantotrichum into pieces or splitting into small pieces, reflux-extracting with 50% ethanol for 5 times each for 2 hr, mixing extractive solutions, recovering ethanol, concentrating to 1:1, adding 10 times of ethanol under stirring, standing for 24 hr, collecting supernatant, recovering ethanol, concentrating to appropriate amount, diluting with water to obtain 60% crude drug solution, filtering, freezing, thawing at room temperature, filtering, diluting to obtain 10% crude drug solution with 30% alcohol content, filtering, and packaging.
Experiment 2 the extraction technique of the invention
Sawing cornu Cervi Pantotrichum into 3-4cm small sections, steaming for 20 min, peeling off skin and hair (to be peeled), and cutting into 1cm thick small pieces.
Placing cornu Cervi Pantotrichum in a hot air circulation oven, setting temperature at 60deg.C, and drying for 8 hr.
Adding cornu Cervi Pantotrichum into extraction tank, extracting under reflux under heating for 5 times, adding 5 times of 50% ethanol each time for 2 hr, mixing extractive solutions 5 times, and concentrating to obtain crude drug: extract 1:1.
adding 95% ethanol with 10 times volume of the concentrated solution under stirring, standing for 24 hr, and collecting supernatant. Recovering ethanol under reduced pressure, concentrating to obtain extract with cornu Cervi Pantotrichum volume of 1/10, diluting with water to obtain 60% solution containing cornu Cervi Pantotrichum decoction pieces, filtering, and freezing. Thawing at room temperature, filtering, diluting to obtain solution containing crude drug 10% and alcohol 30%, and filtering.
Experiment 3 the extraction technique of the invention
Adding 50% glycolysis solution to 5 times of the feeding amount of cornu Cervi Pantotrichum, soaking for 12 hr, timing when the liquid medicine is boiling, and reflux extracting for 4 hr.
Taking out the cornu Cervi Pantotrichum after reflux extraction, and removing skin while it is hot. Adding peeled cornu Cervi Pantotrichum into extraction tank, adding 50% ethanol solution 5 times of the crude drug, reflux-extracting for 4 hr, collecting supernatant each time, mixing the supernatant after reflux-extracting for 5 times, recovering ethanol, and concentrating to obtain medicinal liquid with a ratio of 1:1 with crude drug.
Adding 10 times of 95% ethanol solution into the concentrated solution under stirring, standing for 24 hr after the concentrated solution and 95% ethanol solution are stirred uniformly, collecting supernatant, filtering, concentrating to one tenth of crude drug, discharging, and packaging into clean container. The concentrated solution is weighed, then the water adding amount is calculated, water is added to dilute the concentrated solution into a solution containing 60 percent of crude drug, and the solution is filtered and frozen. Thawing at room temperature, filtering, diluting to obtain solution containing crude drug 10% and alcohol 30%, and filtering.
According to the extraction operation steps of the pantocrine in experiment 3, the velvet antler in different places, different feeding modes (free-range feeding and containment feeding) and different parts are selected for extraction respectively. Firstly, three provinces with better quality of pilose antler are selected: the method comprises the steps of (1) selecting velvet antler collected by different raising modes (free raising and containment) for test in Heilongjiang province, jilin province and Liaoning province; and then taking the velvet antler collected by Heilongjiang province as a raw material, and respectively carrying out extraction tests on the upper part, the middle part and the lower part of the velvet antler.
Identification test detection method
Precisely sucking 1ml of the product, evaporating to dryness in a water bath, dissolving in appropriate amount of water, transferring into a 10ml measuring flask, diluting with water to scale, shaking, taking water as blank, and measuring by spectrophotometry to obtain maximum absorption and minimum absorption at 253+ -2 nm and 232+ -2 nm wavelength, respectively.
Identifying and detecting the pantocrine extracted in the experiments 1-3, comparing the amounts of effective substances in the extracting solutions, and the detection results are shown in table 4; the velvet extract extracted from the selection test of velvet herbs in experiment 4 is detected, and the detection results are shown in tables 5 and 6.
TABLE 4 identification results of pantocrine extracted in experiments 1-3
TABLE 5 identification results of pantocrine extracted from different provinces
TABLE 6 results of identification of pantocrine extracted from different parts of velvet antler in Heilongjiang province
Test results: as can be seen from the experimental data, the pantocrine extracted by the technical scheme of the experiment 3 has a maximum absorption value at the wavelength of 253+/-2 nm and has a minimum absorption value at the wavelength of 232+/-2 nm, and compared with the experiment 1 and the experiment 2, the absorption value is higher; compared with the three-province pilose antler, the pilose antler extract of Jilin province (free-ranging) has the maximum absorption value at the wavelength of 253+/-2 nm and the minimum absorption value at the wavelength of 232+/-2 nm; the cornu Cervi Pantotrichum extract extracted from upper section of cornu Cervi Pantotrichum of Heilongjiang province has maximum absorption at 253+ -2 nm, has minimum absorption at 232+ -2 nm, and has higher maximum absorption value than that extracted from middle section, and has no maximum absorption at 253+ -2 nm and no minimum absorption at 232+ -2 nm. Therefore, the upper section of pilose antler in Jilin province (free-ranging) is the raw material for extracting pilose antler extract.
Example 4
Granulation process
Preferably, the traditional Chinese medicine composition with the effects of tonifying kidney, strengthening spleen, nourishing blood and soothing nerves comprises the following components in parts by weight:
157g of acanthopanax extract, 50ml of schisandra fruit fluid extract, 25ml of 10% pantocrin, 30g of sodium glycerophosphate, a proper amount of calcium carbonate, starch, magnesium stearate and 95% ethanol.
Experiment 1
1. Pretreatment, weighing and batching
Crushing sodium glycerophosphate, sieving with a 100-mesh sieve, sieving calcium carbonate with a 100-mesh vibrating sieve, sieving starch with a 120-mesh sieve, and sieving magnesium stearate with a 100-mesh sieve.
2. Mixing, granulating, and drying
Mixing the weighed raw materials and auxiliary materials with the acanthopanax extract, 10% pantocrin and schisandra chinensis fluid extract, stirring for 20 minutes to uniformly mix the materials, adding a proper amount of 95% ethanol, stirring for 30 minutes to prepare a soft material, granulating by 20 meshes, drying, finishing by a 18-mesh sieve, adding magnesium stearate, mixing for 30 minutes, tabletting, and coating with a film to obtain the traditional Chinese medicine.
Experiment 2
1. Pretreatment, weighing and batching
Crushing sodium glycerophosphate, sieving with a 100-mesh sieve, sieving calcium carbonate with a 100-mesh vibrating sieve, sieving starch with a 120-mesh sieve, and sieving magnesium stearate with a 100-mesh sieve.
2. Mixing, granulating, and drying
Mixing the weighed raw materials and auxiliary materials with the acanthopanax extract and the schisandra chinensis fluid extract, stirring for 20 minutes to uniformly mix the materials, carrying out spray granulation on 10% pantocrine by adopting a one-step granulator, stirring for 10 minutes, then adding a proper amount of 95% ethanol, stirring for 30 minutes to obtain a soft material, granulating by 20 meshes, drying, finishing by a 18-mesh sieve, adding magnesium stearate, mixing for 30 minutes, tabletting, and coating with a film to obtain the traditional Chinese medicine.
Experiment 3
1. Pretreatment, weighing and batching
Crushing sodium glycerophosphate, sieving with a 100-mesh sieve, sieving calcium carbonate with a 100-mesh vibrating sieve, sieving starch with a 120-mesh sieve, and sieving magnesium stearate with a 100-mesh sieve.
2. Mixing, granulating, and drying
Mixing the weighed raw materials with the acanthopanax extract and the schisandra chinensis fluid extract, adding 1/3 of the total amount of 95% ethanol, stirring for 20 minutes to uniformly mix the materials, spraying and mixing 10% pantocrine by adopting a one-step granulator, continuously adding 1/3 of the total amount of 95% ethanol in a spraying manner, stirring for 10 minutes, adding the rest amount of 95% ethanol, stirring for 30 minutes to obtain a soft material, granulating by 20 meshes, drying, finishing by a 18-mesh sieve, adding magnesium stearate, mixing for 30 minutes, tabletting, and coating a film to obtain the finished product.
The acanthopanax extract, the schisandra chinensis fluid extract and 10% pantocrin extract are prepared by adopting the prior art.
1. Pretreatment, weighing and batching
Crushing sodium glycerophosphate, sieving with a 100-mesh sieve, sieving calcium carbonate with a 100-mesh vibrating sieve, sieving starch with a 120-mesh sieve, and sieving magnesium stearate with a 100-mesh sieve.
2. Mixing, granulating, and drying
Mixing the weighed raw materials and auxiliary materials with the acanthopanax extract, 10% pantocrin and schisandra chinensis fluid extract, stirring for 20 minutes to uniformly mix the materials, adding a proper amount of 95% ethanol, stirring for 30 minutes to prepare a soft material, granulating by 20 meshes, drying, finishing by a 18-mesh sieve, adding magnesium stearate, mixing for 30 minutes, tabletting, and coating with a film to obtain the traditional Chinese medicine.
Example 5
A quality study method of Chinese medicinal composition with effects of invigorating kidney, invigorating spleen, nourishing blood and tranquillizing
(1) Content determination of traditional Chinese medicine composition with effects of tonifying kidney, strengthening spleen, nourishing blood and tranquillizing
The method comprises the following steps: extract detection of traditional Chinese medicine composition with effects of tonifying kidney, strengthening spleen, nourishing blood and tranquillizing
Method
Taking 20 pieces of the product, grinding, precisely weighing 2g, placing into a 250ml conical flask, precisely adding 50ml of 60% -80% methanol, sealing, weighing, standing for 1 hour, connecting a reflux condenser, heating to boil, and keeping micro-boiling for 1 hour. After cooling, the conical flask is taken down, the sealing is carried out, the weight is weighed again, the reduced weight is complemented by methanol, shaking is carried out, a drying filter is used for filtering, 25ml of filtrate is precisely measured, the filtrate is placed in an evaporation dish which is dried to constant weight, the evaporation dish is dried in a water bath, the mixture is dried for 3 hours at 105 ℃, and the mixture is cooled in a dryer for 30 minutes, so that the weight is rapidly and precisely weighed. The alcohol-soluble extract content (%) in the test sample was calculated from the dried product unless otherwise specified.
The second method is as follows: method for detecting effective substances in traditional Chinese medicine composition with effects of tonifying kidney, strengthening spleen, nourishing blood and soothing nerves
An Agilent ZB-extension C18 column (250 mmx4.6mm,5 μm) was used; mobile phase A is 0.8% phosphoric acid water solution, mobile phase B is acetonitrile, and the flow rate is 0.8mL/min; the detection wavelength is 220nm; the sample injection amount is 20uL; the column temperature was 40 ℃.
Gradient elution was performed as shown in the following table.
Preparation of reference substance solution
Respectively weighing syringin, eleutheroside E, isofraxidin, schizandrin A, schizandrin B, schizandrin A and schizandrin B reference substances, adding methanol for dissolving to obtain reference substance stock solution with concentration of 1.0mg/mL, precisely sucking appropriate amount of each reference substance stock solution, and adding methanol to obtain mixed solution with concentration of 50 μg/mL. Then respectively and precisely absorbing a proper amount of the control stock solution of the syringin, the eleutheroside E, the isofraxidin, the schizandrin A, the schizandrin B, the schizandrin A and the schizandrin B, and respectively adding methanol to prepare the control solutions with the concentration of 0.2mg/mL, 0.1mg/mL, 0.3mg/mL, 0.1mg/mL and 0.2 mg/mL.
Preparation of test sample solution
Taking 20 pieces of the product, grinding, precisely weighing 2g, precisely weighing, placing in a conical bottle with a plug, precisely adding 50mL of 60% -80% methanol, sealing, weighing the mass, performing ultrasonic treatment for 10min to dissolve, cooling, weighing the mass, supplementing the lost mass with methanol, shaking uniformly, and filtering with a 0.45 microporous filter membrane to obtain a sample solution.
(2) Identification of traditional Chinese medicine composition with effects of tonifying kidney, strengthening spleen, nourishing blood and tranquillizing
A method for identifying a Chinese medicinal composition with effects of invigorating kidney and spleen, nourishing blood and tranquillizing, comprises the following steps:
taking 4 pieces of the product, grinding, precisely weighing 1g of the product, and placing the product in a test tube with a stopper. Adding chloroform 15ml, ultrasonic treating for 20 min, filtering, evaporating filtrate, and dissolving residue with chloroform 1ml to obtain sample solution. And 1g of acanthopanax reference medicine is prepared, and the prepared reference medicine solution is prepared. According to a thin layer chromatography test, 50ul of each of the two solutions is absorbed and respectively spotted on the same silica gel G thin layer plate, and cyclohexane chloroform-ethyl acetate-methanol (10:5:2) is used as developing agent for developing, taken out, dried and inspected under an ultraviolet lamp (365 nm). In the chromatogram of the test sample, fluorescent spots with the same color appear at the positions corresponding to the chromatogram of the reference sample.
The pharmaceutical preparation prepared in example 4 was subjected to the discrimination test according to the above quality test method, and the extract content was measured for each sample in parallel 5 times, and the test results are shown in Table 7.
TABLE 7 extract content detection results
Discrimination test results: the pharmaceutical preparations prepared in the experiment 1, the experiment 2 and the experiment 3 all show fluorescence spots with the same color on the corresponding positions of the chromatogram of the reference substance.
Conclusion: the detection result shows that the drug preparation identification tests prepared in the experiments 1, 2 and 3 in the embodiment 4 all meet the requirements; the average content of the pharmaceutical preparation prepared in the experiment 3 in the extract content detection result is 35.2%, the RSD is 2.4%, and compared with the experiment 1 and the experiment 2, the extract content is uniform, and the active ingredients are uniformly distributed.
Example 6
Clinical trial
1. Patient selection
50 patients with palpitation and insomnia, 25 control groups, 12 men and 13 women, the ages of 30-60 years, and the average age of 40.5 years; treatment group 25, 14 men and 11 women, ages 28-62, average age 42.5 years. The general data for the two groups of patients were not significantly different (p > 0.05) and were comparable.
2. Therapeutic method
The control group was given a powerful Naoqingsu tablet (manufacturer: yabao pharmaceutical company, inc.) for treatment, 3 tablets/time orally, 2 times a day, for treatment for 4 weeks.
The treatment group was given the pharmaceutical composition of the present invention 3 tablets/time orally, 2 times a day, for 4 weeks.
3. Therapeutic effect criterion
And (3) healing: the symptoms of palpitation and insomnia disappear, the night sleep time is more than 6 hours, the deep sleep time is more than 2 hours, and the daytime spirit is full;
the effect is shown: the symptoms of palpitation and insomnia are obviously reduced, the sleeping time is longer than 4 hours, the deep sleeping time is 1-2 hours, and the spirit is improved in the daytime;
the method is effective: the symptoms of palpitation and insomnia are relieved, the sleeping time is increased, and the staged sleeping time is less than 3 hours;
invalidation: the symptoms of palpitation and insomnia are not improved or aggravated.
4. Therapeutic results
The treatment effect is judged according to the sleep quality and the mental state of the patient after curing for 4 weeks, the specific data are shown in the following table 8, the treatment effective rate of the experimental group is 80%, the treatment effective rate of the control group is 56%, and the treatment effective rate of the experimental group is obviously higher than that of the control group.
Table 8 treatment of 2 groups of patients 4 weeks after oral administration
Claims (10)
1. A preparation method of a traditional Chinese medicine composition with the effects of tonifying kidney, strengthening spleen, nourishing blood and tranquillizing is characterized by comprising the following steps:
step one, extraction of acanthopanax extract
Pulverizing radix Acanthopanacis Senticosi into coarse powder, adding 6-8 times of 75% ethanol, reflux extracting for 1-3 hr, and filtering to obtain filtrate 1; extracting the residues for 20-40 min under the assistance of ultrasonic waves at 50-70 ℃ with ultrasonic power of 300W, and filtering to obtain filtrate 2; dissolving the residue in 10 times of water, extracting with microwave for 20-40 min under the assistance of microwave power of 450w, and filtering to obtain filtrate 3; mixing filtrate 1, filtrate 2 and filtrate 3, concentrating, drying, and weighing;
step two, extraction of schisandra chinensis fluid extract
1) Crushing: taking weighted shizandra berry, and putting the shizandra berry into a crusher for crushing;
2) Wetting: placing the weighed coarse schisandra chinensis powder into a clean container, wherein the mass volume ratio is 1kg: 600-700 mL of ethanol with the proportion volume percentage of 40% is uniformly stirred and placed in a closed state for 2 hours;
3) Dipping: placing a filtering device at the bottom of the percolation tank, closing a discharging door, filling the wet schisandra chinensis medicinal material into the percolation tank, covering a layer of 100-mesh silk cloth on the material surface, and firmly pressing the silk cloth by using a stainless steel pore plate; opening a liquid discharge valve below the percolation tank, adding ethanol solution with the volume percentage of 40% from the top of the percolation tank, and discharging air in a decoction piece gap; adding 40% ethanol solution until the liquid medicine flows out from the bottom and the solvent is 2-3cm higher than the surface of the medicinal materials, closing the liquid discharge valve, placing the flowing liquid medicine into the percolating tank, covering the percolating tank with an upper cover, and hermetically soaking for 24h;
4) Percolating: opening a liquid inlet valve and a liquid discharge valve, starting to percolate at the speed of 2-3 mL/kg of medicinal materials per minute, adding ethanol solution with the volume percentage of 40% and the volume of 8 times of the percolate material into the percolate, collecting the primary percolate with the decoction piece quantity of 85%, and finally collecting the continuous percolate until the color is light and nearly colorless for later use;
5) Concentrating: concentrating the percolate of the step 4) to extract of 1.35-1.40;
6) Preparing: cooling the shizandra extract to room temperature, mixing with the primary percolate, stirring uniformly, adding 40% ethanol by volume percent, and adjusting to decoction pieces: fluid extract=1:1, standing for 24h, and collecting supernatant;
step three, extracting the cartialgenous extract with the mass percentage of 10 percent
7) Adding ethanol solution with volume percentage of 50% to 2-10 times of the feeding amount of cornu Cervi Pantotrichum, soaking for 5-24 hr, timing when the liquid medicine boils, and reflux extracting for 2-6 hr;
8) Taking out the cornu Cervi Pantotrichum after reflux extraction, and removing skin while hot; adding peeled cornu Cervi Pantotrichum into extraction tank, adding ethanol solution with volume percentage of 50% 2-10 times of the crude drug, and reflux extracting for 2-6 hr;
9) Repeating step 8) for 2-6 times, collecting supernatant each time, mixing the supernatants after reflux extraction for 2-6 times, recovering ethanol, concentrating cornu Cervi Pantotrichum extractive solution to obtain medicinal liquid with volume ratio of 1:1;
Adding 5-12 times of 95% ethanol solution by volume percentage into the concentrated solution under stirring, standing for 24 hr after the concentrated solution and 95% ethanol solution are stirred uniformly, collecting supernatant, filtering, concentrating to one tenth of crude drug, and packaging into clean container; diluting with water to obtain 60% crude drug solution, filtering, and freezing; thawing at room temperature, filtering, diluting to obtain solution containing crude drug 10% and alcohol 30%, and filtering;
step four, granulating
10 Pre-treatment, weighing and batching
Crushing sodium glycerophosphate, sieving with a 100-mesh sieve, sieving calcium carbonate with a 100-mesh vibrating sieve, sieving starch with a 120-mesh sieve, and sieving magnesium stearate with a 100-mesh sieve;
11 Mixing, granulating, and drying
Mixing the weighed raw materials with the acanthopanax extract extracted in the first step and the schisandra chinensis fluid extract extracted in the second step, adding 1/3 of the total ethanol with the volume percentage of 95%, stirring for 20 minutes to uniformly mix the materials, spraying and mixing the velvet extract with the mass percentage of 10% extracted in the third step by adopting a one-step granulator, continuously adding 1/3 of the total ethanol with the volume percentage of 95% by adopting a spraying mode, stirring for 10 minutes, then adding the ethanol with the residual volume percentage of 95%, stirring for 30 minutes to prepare a soft material, granulating with 20 meshes, drying, sieving with a 18-mesh sieve, adding magnesium stearate, mixing for 30 minutes, tabletting, and coating a film.
2. The method for preparing a Chinese medicinal composition with the effects of tonifying kidney, strengthening spleen, nourishing blood and tranquillizing according to claim 1, wherein the wet shizandra berry medicinal material in the step two 3) is filled into a percolating tank, and the material filling amount is not more than 2/3 of the height of the percolating tank body.
3. The method for preparing a Chinese medicinal composition with kidney-tonifying spleen-strengthening, blood-nourishing and tranquillizing effects according to claim 1, wherein the temperature of the extract in the step two 5) is 50 ℃.
4. The method for preparing a Chinese medicinal composition with the effects of tonifying kidney, strengthening spleen, nourishing blood and tranquillizing according to claim 1, wherein the pilose antler in the step 7) is the upper pilose antler section of the free-ranging of Heilongjiang province.
5. The method for preparing the traditional Chinese medicine composition with the effects of tonifying kidney, strengthening spleen, nourishing blood and tranquillizing according to claim 1, wherein the method is characterized in that in the step 7), the added volume percentage is 5 times of the feeding amount of the 50% glycolysis solution to the pilose antler, after soaking for 12 hours, timing is started when the liquid medicine is boiled, and reflux extraction is carried out for 4 hours.
6. The method for preparing a Chinese medicinal composition with kidney-tonifying spleen-strengthening, blood-nourishing and tranquillizing effects according to claim 1, wherein the step 8) is characterized in that the pilose antler after reflux extraction is taken out and the skin is removed while the pilose antler is hot; adding peeled cornu Cervi Pantotrichum into extraction tank, adding ethanol solution with volume percentage of 50% 5 times of the crude drug, and reflux extracting for 4 hr.
7. The method for preparing a Chinese medicinal composition with effects of invigorating kidney and spleen, nourishing blood and tranquillizing according to claim 1, wherein the step 8) is repeated for 4 times in the step 9), the supernatant is extracted for 5 times each time, the supernatant is concentrated to obtain the cornu Cervi Pantotrichum extract, the ethanol is recovered, and the cornu Cervi Pantotrichum extract is concentrated to the liquid medicine with the volume ratio of 1:1.
8. The method for detecting the extract of the traditional Chinese medicine composition with the effects of tonifying kidney, strengthening spleen, nourishing blood and soothing nerves, which is prepared by the preparation method according to claim 1, is characterized by comprising the following steps:
taking a traditional Chinese medicine composition tablet, grinding, precisely weighing, and placing into a conical flask according to the volume-mass ratio of 2g: adding 60-80% methanol solution in a volume percentage of 50mL, sealing, weighing, standing for 1h, connecting a reflux condenser, heating to boiling, keeping for 1h, cooling, taking down a conical flask, sealing, weighing again, supplementing the reduced weight with methanol, shaking, filtering with a drying filter, precisely measuring 25mL of filtrate, placing into a evaporating dish dried to constant weight, evaporating in a water bath, drying at 105 ℃ for 3h, cooling in a dryer for 30min, precisely weighing; and calculating the content of the alcohol soluble extract in the sample by using the dried product, thus completing the quality detection method.
9. The method for detecting the effective substances of the traditional Chinese medicine composition with the effects of tonifying kidney, strengthening spleen, nourishing blood and soothing nerves according to the preparation method of claim 1 is characterized by comprising the following steps:
1) Preparation of reference substance solution
Respectively weighing syringin, eleutheroside E, isofraxidin, schizandrin A, schizandrin B, schizandrin A and schizandrin B reference substances, respectively adding methanol for dissolving to obtain reference substance stock solutions with concentration of 1.0mg/mL, precisely sucking the reference substance stock solutions, and adding methanol to obtain mixed solution with concentration of 50 μg/mL; respectively and precisely absorbing the control stock solutions of syringin, eleutheroside E, isofraxidin, schizandrin A, schizandrin B, schizandrin A and schizandrin B, and adding methanol to prepare control solutions with the concentration of 0.2mg/mL, 0.1mg/mL, 0.3mg/mL, 0.1mg/mL and 0.2 mg/mL;
2) Preparation of test sample solution
Taking a traditional Chinese medicine composition tablet with the effects of tonifying kidney and strengthening spleen, nourishing blood and tranquillizing, grinding, precisely weighing 2g, precisely weighing, placing in a conical bottle with a plug, precisely adding 50mL of 60% -80% methanol, sealing, weighing the mass, performing ultrasonic treatment for 10min to dissolve, cooling, weighing the mass, supplementing the lost mass with methanol, shaking uniformly, and filtering with a 0.45 microporous filter membrane to obtain a sample solution;
An Agilent ZB-Extend C18 chromatographic column is adopted, 250mmx4.6mm,5 mu m; mobile phase A is 0.8% phosphoric acid water solution, mobile phase B is acetonitrile, and the flow rate is 0.8mL/min; the detection wavelength is 220nm; the sample injection amount is 20uL; the column temperature is 40 ℃, and gradient elution is carried out according to the following mode, namely the detection of the effective substances is completed;
10. the method for identifying the traditional Chinese medicine composition with the effects of tonifying kidney, strengthening spleen, nourishing blood and soothing nerves by the preparation method as claimed in claim 1 is characterized by comprising the following steps:
grinding the Chinese medicinal composition tablet, and placing in a test tube with a stopper; the volume mass ratio is 1g: adding chloroform into 15mL, performing ultrasonic treatment for 20min, filtering, evaporating filtrate to dryness, and adding chloroform into residue 1mL to dissolve to obtain sample solution; preparing 1g of acanthopanax reference medicine, and preparing a reference medicine solution by the same method; according to thin layer chromatography test, sucking 50ul of each of the above two solutions, respectively spotting on the same silica gel G thin layer plate, developing with cyclohexane chloroform-ethyl acetate-methanol as developing agent, taking out, air drying, inspecting under 365nm ultraviolet lamp, and displaying fluorescent spots of the same color on the position corresponding to the reference sample chromatogram in the sample chromatogram; wherein the volume ratio of petroleum ether to normal hexane to ethyl acetate to methanol is 8:10:3:2.
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| CN1768762A (en) * | 2005-08-08 | 2006-05-10 | 北京阜康仁生物制药科技有限公司 | Novel drug administering route of pantocrine injection, its preparation process and novel indications |
| CN101322570A (en) * | 2007-06-11 | 2008-12-17 | 哈尔滨完达山药业有限公司 | Process for preparing acanthopanax root drink |
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| CN1768762A (en) * | 2005-08-08 | 2006-05-10 | 北京阜康仁生物制药科技有限公司 | Novel drug administering route of pantocrine injection, its preparation process and novel indications |
| CN101322570A (en) * | 2007-06-11 | 2008-12-17 | 哈尔滨完达山药业有限公司 | Process for preparing acanthopanax root drink |
| CN101618113A (en) * | 2008-06-30 | 2010-01-06 | 广州白云山和记黄埔中药有限公司 | Traditional Chinese medicine and western medicine composition containing fine powder and preparation method thereof |
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