New method for extracting and concentrating high-efficiency components of kidney-tonifying Juanbi pill

Abstract

The invention discloses a novel method for extracting and concentrating efficient components of a kidney-tonifying Juanbi pill, which comprises the following steps: pretreating medicinal materials: taking prescription medicinal materials of the kidney-tonifying Juanbi pill; the traditional Chinese medicine composition is prepared from rhizoma drynariae, prepared rehmannia root, Chinese angelica, paniculate swallowwort root, ground beetle, stiff silkworm (stir-fried with bran), centipede, scorpio, honeycomb (stir-fried with clear water), lumbricus (processed with wine), zaocys dhumnade (processed with wine), corydalis tuber, pyrola, epimedium, berba aristolochiae mollissimae, geranium wilfordii, caulis spatholobi, scandent hop, radix rehmanniae recen and polygonum cuspidatum. Through targeted medicinal material pretreatment and ultrasonic-assisted extraction, the problems of slow dissolution of plant medicinal effective components and difficult extraction of animal medicinal cuticle of the kidney-tonifying Juanbi pill are effectively solved, and the extraction rate of the medicinal effective components stably reaches 60% or above. Effective components of medicinal materials such as polygonum cuspidatum, caulis spatholobi, berba aristolochiae mollissimae, geranium wilfordii and rehmannia in the prescription can be fully dissolved out, and waste of raw materials is avoided.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of component extraction and concentration, and particularly relates to a new method for high-efficiency component extraction and concentration of Yishen Ju Bi Pills. BACKGROUND

[0002] Currently, in the field of production process of Yishen Ju Bi Pills, for the processing of its prescription medicinal materials (including plant medicines such as Psammosilene tunicoides, Radix Rehmanniae Preparata, Angelica sinensis, and animal medicines such as Eupolyphaga, centipede, and Scorpio), the existing technology usually includes the following links: medicinal material pretreatment, effective component extraction, extraction liquid purification, concentration, and subsequent preparation adaptation. Among them, the medicinal material pretreatment usually adopts a conventional crushing method to crush the cleaned and impurity-removed medicinal materials to a particle size of 80-120 meshes; the effective component extraction mainly adopts a hot reflux extraction method, which controls a certain temperature and time to realize component dissolution by adding an ethanol aqueous solution as a solvent; the extraction liquid purification usually adopts a micro-pore filter membrane to remove solid residues; the concentration link generally uses a reduced pressure concentration process to reduce the concentration temperature and the loss of part of the components by reducing the environmental pressure; and then, the concentrated liquid is dried into a dry extract by a conventional drying method, and is mixed with auxiliary materials to prepare a pill, a tablet, or a capsule, etc. dosage form according to the needs to meet the clinical application needs.

[0003] Although the existing Yishen Ju Bi Pills production process can realize product preparation, it has obvious deficiencies in combination with the core beneficial effects of the present method: first, the effective component extraction efficiency is limited, the conventional crushing method is difficult to break the plant medicine cell wall, and the hot reflux extraction method cannot adapt to the differentiated dissolution needs of animal medicines and plant medicines, resulting in slow dissolution of plant medicine effective components, insufficient extraction of animal medicine effective components, and difficulty in achieving high extraction rate; second, the retention effect of heat-sensitive components is poor, the temperature of the hot reflux extraction (usually 60-80℃) and the reduced pressure concentration (usually 45-55℃) is still high, which easily causes the degradation of heat-sensitive active components such as alkaloids and active peptides; third, the impurity removal capacity is insufficient, the micro-pore filter membrane filtration cannot remove small molecule impurities such as monosaccharides and inorganic salts, which easily leads to low purity of the concentrated liquid and may cause equipment blockage problems in the subsequent process; fourth, the energy consumption is high and the production efficiency is low, the hot reflux extraction time is long (usually 2-4h), and the reduced pressure concentration energy consumption is large, which does not meet the industrialization needs of high efficiency and energy saving; fifth, the product stability and preparation adaptability need to be improved, the dry extract prepared by the existing drying process is easy to absorb moisture, and the auxiliary material ratio is also difficult to balance the preparation stability and disintegration performance. SUMMARY

[0004] In order to overcome the above-mentioned defects of the prior art, the present application provides a new method for high-efficiency component extraction and concentration of Yishen Ju Bi Pills, which solves the problems of low extraction rate, large loss of heat-sensitive components, many impurities, high energy consumption, and poor stability in the prior art Yishen Ju Bi Pills process.

[0005] To achieve the above-mentioned purposes, the present application provides the following technical solutions: The new method for extracting and concentrating the high-efficiency components of Yishen Ju Bi Pills comprises the following steps: Medicine pretreatment: Take the prescription medicines of Yishen Ju Bi Pills, which are composed of rhizoma drynariae, radix rehmanniae preparatae, angelica sinensis, radix clematidis, eupolyphaga, scolopendra subthecalis, scorpio, flos farinae (stir-fried), cordyline frutescens (wine processed), rhabdophis nigrocinctus (wine processed), corydalis, pedunculate eriobotrya, epimedium, radix rubiae, old stork grass, caulis broussonetiae, humulus scandens, radix rehmanniae, giant knotweed; wherein, the preparation conditions of silkworm chrysalis (wheat bran fried) are as follows: 5%-8% of wheat bran is added according to the mass of the medicine, the frying temperature is controlled at 120-140 ℃, and the frying is continued for 5-8 min; the preparation conditions of flos farinae (stir-fried) are as follows: the frying temperature is controlled at 110-130 ℃, and the frying is continued for 3-5 min; the preparation conditions of cordyline frutescens (wine processed) and rhabdophis nigrocinctus (wine processed) are as follows: after being soaked in 15%-20% of yellow rice wine for 2-3 h, they are dried at 50-60 ℃; the above medicines are cleaned and impurities are removed, and then they are mixed and crushed to 80-120 mesh, to obtain the medicine powder; (2) Ultrasonic-assisted extraction: the medicine powder obtained in step (1) is added into an extraction tank, 30%-70% of ethanol aqueous solution is added as the extraction solvent according to the solid-liquid ratio of 1:8-1:15, the ultrasonic-assisted extraction device is started, the ultrasonic power is controlled at 200-600 W, the extraction temperature is controlled at 40-65 ℃, and the extraction time is controlled at 30-90 min, and the extraction is completed to obtain the extraction liquid; (3) Filtration of the extraction liquid: the extraction liquid obtained in step (2) is filtered under reduced pressure by using a 0.22-0.45 μm microporous filter membrane, and the clear extraction liquid is collected after the filter residue is removed; (4) Concentration by nanofiltration membrane: the clear extraction liquid obtained in step (3) is introduced into a nanofiltration membrane separation system, the material of the nanofiltration membrane is polyethersulfone or sulfonated polyethersulfone, the operation pressure is controlled at 1.0-3.0 MPa, and the concentration temperature is controlled at 25-40 ℃ for low-temperature concentration treatment, until the relative density of the extraction liquid at 25 ℃ reaches 1.10-1.30 g / cm³, to obtain the concentrated liquid of Yishen Ju Bi Pills; In the method, the extraction rate of the effective components of the medicine is ≥60%, and the retention rate of the heat-sensitive active components, including alkaloids and active peptides, is not less than 90%.

[0006] Preferably, in step (1), the particle size of the crushed medicine is 100 mesh, and the environment temperature during the crushing process is controlled at not more than 30 ℃ and the relative humidity is controlled at not more than 60%.

[0007] Preferably, in step (2), the solid-liquid ratio is 1:12, the volume fraction of the ethanol aqueous solution is 50%, the ultrasonic power is 400 W, the extraction temperature is 55 ℃, and the extraction time is 60 min.

[0008] Preferably, in step (2), the ultrasonic-assisted extraction process adopts an intermittent ultrasonic mode, i.e. after 30s of ultrasonic operation, pause for 10s, and repeat the operation until the set extraction time is reached.

[0009] Preferably, in step (4), the molecular weight cut-off of the nanofiltration membrane is 100-500 Da, and the nanofiltration membrane is cleaned online every 30 min during the concentration process, and the cleaning solution is 0.5%-1.0% sodium hydroxide aqueous solution.

[0010] Preferably, it further comprises step (5) post-treatment of the concentrated solution: the concentrated solution of Yishen Ju Bi Wan obtained in step (4) is placed in a vacuum drying box, the vacuum degree is controlled at-0.08 to-0.10 MPa, and the drying temperature is controlled at 50-60℃, and the moisture content is dried to ≤5%, to obtain a dry extract of Yishen Ju Bi Wan, which is used to prepare pills.

[0011] Preferably, the pretreatment of medicinal materials in step (1) further comprises an enzymatic hydrolysis step: the cleaned and impurity-removed Rhizoma Drynariae, Radix Rehmanniae Praeparatae, Angelica sinensis, Cyrtomium fortunei, Corydalis, Cornus officinalis, Epimedium, Rabdosia lophanthoides, Old Stork Grass, Millettia nitida, Humulus scandens, Rehmannia glutinosa, Giant Knotweed, and Bombyx Batryticatus (roasted with wheat bran) are ground to 40-60 mesh, 0.1%-0.3% cellulase solution with an enzyme activity of 5000 U / g is added, the temperature is controlled at 45-55℃, and the pH is controlled at 4.5-5.5, and the enzymatic hydrolysis is carried out for 1-2h; after enzymatic hydrolysis, the above-mentioned medicinal materials are mixed with the previously ground Eupolyphaga, Solpugid, Scorpio, Fungus Honeycomb (fried), Giant Salamander (wine), and Rhabdophis (wine) to 80-100 mesh, and then the mixture is ultra-micro ground to 150-200 mesh to obtain a medicinal material powder; the combination of enzymatic hydrolysis and ultra-micro grinding shortens the extraction time of effective components of plant medicinal materials such as Rhizoma Drynariae and Radix Rehmanniae Praeparatae by 15%-25%, and the loss of effective components during the grinding process is ≤3%.

[0012] Preferably, the ultrasonic-assisted extraction in step (2) adopts a phased control: in the first phase, only the powders of Eupolyphaga, Solpugid, Scorpio, Fungus Honeycomb (fried), Giant Salamander (wine), and Rhabdophis (wine) are subjected to ultrasonic extraction, the ultrasonic power is controlled at 500-600W, the temperature is controlled at 55-65℃, and the time is controlled at 40-50min; in the second phase, the powders of Rhizoma Drynariae, Radix Rehmanniae Praeparatae, Angelica sinensis, Cyrtomium fortunei, Corydalis, Cornus officinalis, Epimedium, Rabdosia lophanthoides, Old Stork Grass, Millettia nitida, Humulus scandens, Rehmannia glutinosa, and Giant Knotweed are added to the extraction tank, the ultrasonic power is adjusted to 300-400W, the temperature is adjusted to 40-50℃, and the time is adjusted to 20-40min to complete the extraction; the phased extraction increases the extraction rate of insect active peptides in animal medicinal materials such as Eupolyphaga and Solpugid by ≥55%, and reduces the dissolution amount of polysaccharide small molecule impurities in plant medicinal materials such as Rhizoma Drynariae and Radix Rehmanniae Praeparatae by 10%-15%.

[0013] Preferably, between step (3) and step (4), there is also step (3a) impurity adsorption: the clear extract obtained in step (3) is passed into a resin column filled with D101 type or AB-8 type macroporous adsorption resin, the flow rate is controlled at 1-2 BV / h, and the temperature is controlled at 25-30 DEG C; after adsorption is completed, the eluate is collected after elution with 10%-20% ethanol aqueous solution; the eluate is used as the feed liquid for nanofiltration membrane concentration in step (4); the macroporous adsorption resin can remove 80%-90% of monosaccharides and inorganic salts in the extract, and the recovery rate of alkaloids and active peptides in Yishen Ju Bi pills is greater than or equal to 95%.

[0014] Preferably, in step (5), the obtained Yishen Ju Bi pill dry extract is crushed to a particle size of 120-150 meshes to obtain a dry extract powder; when the dry extract powder is used to prepare pills, the dry extract powder and the auxiliary materials are mixed at a mass ratio of 1:0.3-0.8, and the auxiliary materials include starch and microcrystalline cellulose.

[0015] The technical effects and advantages of the new Yishen Ju Bi pill efficient ingredient extraction and concentration method are as follows: 1. The method significantly improves the extraction efficiency of effective ingredients, and is suitable for complex medicinal material compositions by targeted medicinal material pretreatment and ultrasonic-assisted extraction, effectively solving the problems of slow dissolution of plant medicine effective ingredients and thick cutin layer of animal medicines in Yishen Ju Bi pills, so that the extraction rate of medicinal material effective ingredients is stably above 60%, and the effective ingredients of plant medicines such as Drynariae Rhizoma and Rehmanniae Radix Praeparata and animal medicines such as Eupolyphaga and Centipede in the prescription can be fully dissolved, thereby avoiding waste of raw materials.

[0016] 2. The method controls the temperature at 40-65 DEG C in ultrasonic-assisted extraction and controls the temperature at 25-40 DEG C in nanofiltration membrane concentration, thereby avoiding damage to heat-sensitive components such as alkaloids and active peptides in Yishen Ju Bi pills in traditional high-temperature processes, retaining the retention rate of heat-sensitive active ingredients to be not less than 90%, ensuring the content of effective ingredients of the final product, and avoiding the decrease of drug efficacy caused by degradation of components.

[0017] 3. The method can remove 80%-90% of small molecular impurities such as monosaccharides and inorganic salts in the extract through the macroporous adsorption resin impurity adsorption link, while ensuring that the recovery rate of effective components such as alkaloids and active peptides is not less than 95%, which not only improves the purity of Yishen Ju Bi pill concentrated solution, but also reduces the membrane clogging problem in the subsequent nanofiltration membrane concentration process, maintains the stable operation efficiency of the nanofiltration system, and avoids the decrease of concentration efficiency or the non-standard product purity caused by impurity enrichment.

[0018] 4. This invention shortens the extraction time (30-90 min) through the ultrasonic cavitation effect, and combined with the high-efficiency concentration characteristics of nanofiltration membranes, it significantly reduces heating energy consumption and production time. At the same time, the optimization of the enzymatic hydrolysis-ultra-fine grinding in the pretreatment stage and the staged ultrasonic extraction in the extraction stage further improves the efficiency of each stage, significantly reduces unit energy consumption, and shortens the production cycle.

[0019] 5. This invention expands the application scenarios. The dry extract obtained after the concentrated liquid is subsequently vacuum dried (moisture content ≤5%) has small fluctuations in the content of effective ingredients and strong stability during storage. The dry extract can be prepared into various dosage forms such as pills, tablets, and capsules by reasonable excipient ratio. Attached Figure Description

[0020] Figure 1 This is a flowchart of a novel method for extracting and concentrating highly efficient components of Yishen Juanbi Pills proposed in this invention. Detailed Implementation

[0021] The technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings of the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention.

[0022] It should be noted that, in this document, relational terms such as "first" and "second" are used only to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include," "contain," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that includes a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Without further limitation, an element defined by the phrase "includes..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes said element.

[0023] refer to Figure 1 This invention provides a novel method for the extraction and concentration of highly efficient components of Yishen Juanbi Pills. The core technical solution revolves around the prescription herbs of Yishen Juanbi Pills (Drynaria fortunei, Rehmannia glutinosa, Angelica sinensis, Cynanchum paniculatum, Eupolyphaga sinensis, Bombyx mori (stir-fried with wheat bran), Scolopendra subspinipes, Buthus martensii, Vespa nidus (stir-fried), Pheretima aspergillum (processed with wine), Zaocys dhumnades (processed with wine), Corydalis yanhusuo, Pyrola rotundifolia, Epimedium brevicornu, Clematis chinensis, Geranium wilfordii, Spatholobus suberectus, Humulus scandens, Rehmannia glutinosa, Polygonum cuspidatum). The specific steps include the following steps, which can be optimized according to specific needs: Core process steps: Medicinal material pretreatment: take the above prescription medicinal materials, complete the processing according to the specific requirements: 5%-8% bran, 120-140℃, 5-8min; honeycomb (stir-fried): 110-130℃, 3-5min; broad earthworm (wine), wine: 15%-20% yellow wine soak 2-3h, 50-60℃ drying); after cleaning and removing impurities, all medicinal materials are mixed and pulverized to 80-120 mesh to obtain medicinal material powder; the environmental temperature during the pulverization process is controlled to be ≤30℃, and the relative humidity is ≤60%, so as to ensure the stability of the medicinal material components.

[0024] Ultrasonic-assisted extraction: the medicinal material powder is added into the extraction tank, 30%-70% ethanol aqueous solution is added as the extraction solvent according to the solid-liquid ratio of 1:8-1:15, the ultrasonic device is started, the ultrasonic power is controlled to be 200-600W, the extraction temperature is controlled to be 40-65℃, and the extraction time is controlled to be 30-90min, so as to complete the extraction to obtain the extraction liquid; during the extraction process, the "intermittent ultrasonic mode" (ultrasonic 30s, pause 10s, cycle) can be used, or the medicinal materials are extracted in stages according to the types of medicinal materials (animal medicines: 500-600W, 55-65℃, 40-50min; plant medicines: 300-400W, 40-50℃, 20-40min), which is suitable for the extraction requirements of different medicinal materials.

[0025] Extraction liquid filtration: the extraction liquid is filtered under reduced pressure by using a 0.22-0.45μm microporous filter membrane, and the clear extraction liquid is collected after removing the filter residue, so as to avoid the influence of solid impurities on the subsequent process.

[0026] Nanofiltration membrane concentration: the clear extraction liquid is introduced into a nanofiltration membrane separation system (membrane material is polyether sulfone or sulfonated polyether sulfone, molecular weight cutoff is 100-500Da), low-temperature concentration is carried out by controlling the operating pressure to be 1.0-3.0MPa and the concentration temperature to be 25-40℃, until the relative density of the extraction liquid at 25℃ reaches 1.10-1.30g / cm³, and the Yishen Ju Bi pill concentrated solution is obtained; during the concentration process, the nanofiltration membrane can be cleaned online every 30min by using 0.5%-1.0% sodium hydroxide aqueous solution, so as to prevent the membrane from being blocked and ensure the concentration efficiency.

[0027] Optional optimization steps: Pretreatment optimization (enzyme hydrolysis-ultrafine pulverization combination): the plant medicinal materials (drynaria, rehmannia glutinosa, etc.) are first pulverized to 40-60 mesh, 0.1%-0.3% cellulase solution with an enzyme activity of 5000U / g is added, and enzyme hydrolysis is carried out at 45-55℃ and pH4.5-5.5 for 1-2h; after enzyme hydrolysis, the animal medicinal materials pulverized to 80-100 mesh are mixed, and then ultrafine pulverization is carried out to 150-200 mesh, so as to further improve the dissolution efficiency of the effective components of the plant medicines.

[0028] Impurity adsorption optimization: Add a "macroporous adsorption resin impurity removal" step between filtration and concentration. Pass the clarified extract into a D101 or AB-8 macroporous adsorption resin column, control the flow rate at 1-2 BV / h, and the temperature at 25-30°C. After adsorption, elute with 10%-20% ethanol aqueous solution, and collect the eluate for subsequent concentration. This can remove 80%-90% of monosaccharides and inorganic salts, and ensure that the effective ingredient recovery rate is ≥95%.

[0029] Concentrate post-treatment and preparation adaptation: Place the concentrate in a vacuum drying oven (vacuum degree -0.08 to -0.10 MPa, temperature 50-60°C), dry to a moisture content of ≤5%, and obtain the dry extract of Yishen Ju Bi pills. The dry extract can be crushed to 120-150 mesh, and auxiliary materials such as starch and microcrystalline cellulose are added at a mass ratio of dry extract powder: auxiliary material = 1:0.3-0.8. After mixing, prepare pills, tablets, or capsules to meet different preparation needs.

[0030] Through the combination of the above core process and optional optimization steps, the effective ingredient extraction rate of medicinal materials can be ≥60%, and the retention rate of heat-sensitive active ingredients can be ≥90%, meeting the quality and production requirements of Yishen Ju Bi pills.

[0031] General basic information: Medicinal material preparation: Process medicinal materials according to the requirements of the Yishen Ju Bi pill prescription, such as stir-frying silkworm chrysalis (with bran) with 6% bran of the medicinal material weight at 130°C for 6 minutes, stir-frying bee nest at 120°C for 4 minutes, and soaking earthworms and rat snakes in 18% yellow rice wine for 2.5 hours and then drying at 55°C. Other medicinal materials are cleaned and impurities are removed according to the prescription requirements.

[0032] Detection method: Focus on measuring three key indicators: 1) the proportion of effective ingredients (angelica-derived ferulic acid and corydalis-derived corydaline) dissolved, measured by high-performance liquid chromatography; 2) the proportion of heat-sensitive active ingredients (alkaloids and active peptides) retained, measured by ultraviolet spectrophotometry; and 3) the content of impurities such as monosaccharides and inorganic salts, measured by ion chromatography.

[0033] Core equipment: ultrasonic extraction machine with adjustable power (power range 0-1000W), dedicated nanofiltration membrane equipment (membrane material is polyether sulfone or sulfonated polyether sulfone), and vacuum drying oven (vacuum degree up to -0.08 to -0.10 MPa).

[0034] Example 1 This example presents a new method for extracting and concentrating high-efficiency ingredients of Yishen Ju Bi pills, which is used for the basic extraction and concentration process of Yishen Ju Bi pills. The specific implementation content includes: Implementation purpose: Verify the feasibility of the core process from the processing of medicinal materials to ultrasonic extraction, filtration and residue removal, and membrane concentration. Also test the effects of the parameters such as 1:12 liquid-to-solid ratio, 50% alcohol solution, and 400W ultrasonic power during extraction, as well as the stability of the nanofiltration membrane (molecular weight cut-off 300Da, online cleaning every 30 minutes) during concentration.

[0035] Implementation steps: (1) Pre-treatment of medicinal materials: Take the medicinal materials in the prescription (drynaria 106g, rehmannia 132g, angelica 106g, xuchangqing 106g, eupolyphaga 106g, silkworm 106g, scolopendra 16g, centipede 16g, honeycomb 106g, radix litseae 106g, snake 106g, corydalis 106g, pedunculate 106g, epimedium 106g, old crane grass 132g, chicken blood 132g, hop 132g, raw rehmannia 185g, giant knotweed 132g), clean and remove impurities, then mix and crush to 100 mesh. Control the environment temperature at 28℃ and humidity at 55% during crushing. Get the fine powder of medicinal materials.

[0036] (2) Ultrasonic extraction of active ingredients: Put the fine powder of medicinal materials into the extraction tank, add solvent according to the ratio of 1 part of medicinal materials to 12 parts of 50% alcohol solution, turn on the ultrasonic extractor, adjust the power to 400W, the temperature to 55℃, and extract for 60 minutes. Get the solution containing active ingredients after extraction.

[0037] (3) Filtration and residue removal: Filter the extracted solution under reduced pressure with a 0.3μm fine filter (range between 0.22-0.45μm) to remove the undissolved medicinal material residue, leaving only the clear solution.

[0038] (4) Membrane concentration of medicinal liquid: Put the clear solution into the nanofiltration membrane equipment (use polyether sulfone membrane, molecular weight cut-off 300Da, range 100-500Da), control the operating pressure at 2.0MPa (range 1.0-3.0MPa) and the temperature at 30℃ (range 25-40℃), clean the membrane online with 0.8% sodium hydroxide solution (range 0.5%-1.0%) every 30 minutes. Continue to concentrate until the relative density of the solution reaches 1.20g / cm³ (range 1.10-1.30g / cm³) at 25℃. Get the concentrated solution of Yishen Ju Bi Wan.

[0039] Implementation effect: The dissolution ratio of active ingredients is 65.3%, which meets the requirements; The heat-sensitive alkaloids are retained at 92.5% and the active peptides are retained at 91.8%, without obvious damage, ensuring the efficacy; The nanofiltration membrane continuously runs for 2 hours without clogging, and the concentration efficiency is stable, which can meet the continuous production demand.

[0040] Example 2 This example proposes a new method for extracting and concentrating the high-efficiency components of Yishen Ju Bi Pills, which is used for the enzymatic hydrolysis and ultra-fine grinding optimization of medicinal materials in Yishen Ju Bi Pills. The specific implementation content includes: Implementation purposes: The use of cellulase to break down plant medicine cell walls and then ultra-fine grinding solves the problem of slow dissolution of plant medicine active ingredients, and is used in conjunction with the core process.

[0041] Implementation steps: (1) Medicinal material pre-treatment: Plant medicine enzymatic hydrolysis: Take the plant medicines in the prescription (drynaria 106g, rehmannia glutinosa 132g, angelica 106g, xuchangqing 106g, corydalis 106g, pyrola 106g, epimedium 106g, old gong 106g, old gong 132g, chicken blood 132g, hemp 132g, radix rehmanniae 185g, giant knotweed 132g), after cleaning and removing impurities, crush to 50 mesh (range 40-60 mesh), add cellulase solution with mass fraction 0.2%, enzyme activity 5000u / g, control temperature 50℃ (range 45-55℃), pH 5.0 (range 4.5-5.5), enzymatic hydrolysis for 1.5 hours (range 1-2 hours); Animal medicine grinding: Take the animal medicines in the prescription (ground beetle 106g, centipede 16g, scorpion 16g, honeycomb (fried) 106g, broad earth dragon (wine) 106g, rink snake (wine) 106g, silkworm (wheat) 106g), after cleaning and removing impurities, crush to 90 mesh (range 80-100 mesh); Mix and ultra-fine grind: Mix the enzymatically hydrolyzed plant medicines and the ground animal medicines together, and ultra-fine grind to 180 mesh (range 150-200 mesh) to obtain fine medicinal powder.

[0042] (2) Ultrasonic extraction of active ingredients: Add solvent according to the ratio of 1 part of medicinal material to 10 parts of 45% alcohol solution, ultrasonic power 350w, temperature 50℃, extraction time 45 minutes, to obtain the extract.

[0043] (3) Filtration and residue removal + membrane concentration: As in Example 1, filter with a 0.3μm fine filter, then concentrate with polyether sulfone membrane (300Da), 2.0MPa pressure, 30℃ temperature, until the relative density of the solution is 1.18g / cm³ at 25℃.

[0044] Implementation effect: The extraction time of plant medicine active ingredients is shortened by 20% compared with Example 1, solving the problem of slow dissolution of plant medicine. The loss of effective components during the crushing process is only 2.2%, and there is no waste of raw materials; The total dissolution ratio of effective components is 68.7%, and the retention of heat-resistant components is 93.1%, further improving the process effect.

[0045] Example 3 This example proposes a new method of extracting and concentrating high-efficiency components of Yishen Ju Bi Pills, which is used for optimizing the phased ultrasonic extraction of Yishen Ju Bi Pills (adapted to animal medicines and plant medicines). The specific implementation content includes: Implementation purposes: Verify the effect of phased ultrasonic extraction - because animal medicines (such as earthworms and centipedes) have thick skins and are difficult to extract, the extraction conditions of plant medicines are different from those of animal medicines, so the ultrasonic parameters are adjusted separately to improve the dissolution ratio of effective components of animal medicines and reduce the dissolution of impurities in plant medicines, which are used in conjunction with the core process.

[0046] Implementation steps: (1) Pre-treatment of medicinal materials: Take the same prescription medicinal materials as in Example 1, and after cleaning and removing impurities, crush them separately: animal medicines (5g of earthworms and seven other kinds) and plant medicines (10g of drynaria and 13 other kinds) are crushed to 100 mesh to obtain two kinds of fine powder.

[0047] (2) Phased ultrasonic extraction of effective components: First stage extraction of animal medicines: only animal medicine fine powder is put into the extraction tank, and the solvent is added at a ratio of 1 part of medicine to 12 parts of 50% alcohol solution, the ultrasonic power is 550W (range 500-600W), the temperature is 60℃ (range 55-65℃), and the extraction time is 45 minutes (range 40-50 minutes); Second stage extraction of plant medicines: add plant medicine fine powder to the extraction tank, adjust the ultrasonic power to 350W (range 300-400W), the temperature to 45℃ (range 40-50℃), and the extraction time to 30 minutes (range 20-40 minutes), and the extraction is completed to obtain the extraction liquid.

[0048] (3) Filtration and impurity removal + membrane concentration: As in Example 1, filter with a 0.3μm fine filter, then concentrate with a polyether sulfone membrane (300Da), 2.0MPa pressure, and 30℃ temperature, until the relative density of the solution is 1.22g / cm³ at 25℃.

[0049] Implementation effect: The dissolution ratio of insect active peptides in animal medicines is 57.2%, which is 8.6 percentage points higher than that of mixed ultrasonic extraction in Example 1, ensuring the sufficient dissolution of effective components of animal medicines; The dissolution amount of polysaccharide impurities in plant medicines is reduced by 13.5% compared with Example 1, reducing the pressure of subsequent impurity treatment; The total effective component dissolution ratio is 66.5%, and the heat component retention is 92.8%, which ensures the extraction efficiency and controls the product purity.

[0050] Embodiment 4 This embodiment proposes a new method for extracting and concentrating the high-efficiency components of Yishen Ju Bi Pills, which is used for removing impurities (excluding monosaccharides and inorganic salts) from Yishen Ju Bi Pills with macroporous adsorption resin. The specific implementation content includes: Implementation purpose: Verify the effect of removing impurities with macroporous adsorption resin. Before membrane concentration, remove the useless impurities such as monosaccharides and inorganic salts in the concentrated solution, while ensuring that the effective components are not lost, and cooperate with the core process.

[0051] Implementation steps: (1) Pre-treatment of medicinal materials + ultrasonic extraction + filtration to remove residue: As in Embodiment 1: grind the medicinal materials to 100 mesh, extract at 400W ultrasonic power and 55℃ for 60 minutes, and filter with a 0.3μm fine filter to obtain a clear extract.

[0052] (2) Remove impurities with resin: Pass the clear extract into a column filled with D101 type macroporous adsorption resin (AB-8 type can also be used), control the solution flow rate at 1.5BV / h (range 1-2BV / h), and the temperature at 28℃ (range 25-30℃). After adsorption is complete, elute with a 15% alcohol solution (range 10%-20%) and collect the eluted solution.

[0053] (3) Membrane concentration of medicinal liquid: Pass the eluted solution into a nanofiltration membrane device (as in Embodiment 1), and concentrate to a solution relative density of 1.20g / cm³ at 25℃ to obtain Yishen Ju Bi Pill concentrate.

[0054] Implementation effect: 86.3% of the monosaccharides and 88.5% of the inorganic salts in the extract are removed, and the purity of the concentrate is improved by 18.2% compared to Embodiment 1, meeting the product purity requirements; The alkaloid recovery rate is 95.7%, and the active peptide recovery rate is 96.2%, with no significant loss of effective components; The subsequent nanofiltration membrane clogging rate is reduced, and the concentration time is shortened by 15% compared to Embodiment 1, improving the overall process efficiency.

[0055] Embodiment 5 This embodiment proposes a new method for extracting and concentrating the high-efficiency components of Yishen Ju Bi Pills, which is used for Yishen Ju Bi Pill concentrate subsequent processing (making dry extract) and adapting to preparation (making pills) verification. The specific implementation content includes: Implementation purpose: To verify the effect of the subsequent processing of the concentrated liquid into dry extract, and the effect of the dry extract plus auxiliary materials into pills, which is the requirement of YiShenJieBi Wan, and is used in conjunction with the core process.

[0056] Implementation steps: (1) Extraction + membrane concentration: As in Example 1, complete the processing of medicinal materials, ultrasonic extraction, filtration and residue removal, and membrane concentration to obtain YiShenJieBi Wan concentrated liquid.

[0057] (2) Subsequent processing of concentrated liquid into dry extract: Put the concentrated liquid into a vacuum drying oven, control the vacuum degree to -0.09 MPa, and the temperature to 55°C (range 50-60°C), dry to a moisture content of 4.2% (requirement ≤5%), to obtain YiShenJieBi Wan dry extract.

[0058] (3) Dry extract adapted to make pills: Grind the dry extract to 130 mesh (range 120-150 mesh), add the mixed auxiliary materials of starch and microcrystalline cellulose at a mass ratio of dry extract powder to auxiliary materials 1:0.5 (range 1:0.3-0.8), mix evenly, and then make into pills.

[0059] Implementation effect: The moisture content of the dry extract is 4.2%, and there is no moisture absorption and deterioration after 3 months of storage, ensuring the stability of the product storage; The content of ferulic acid in the dry extract is 0.85 mg / g, and the content of fumarolic acid is 0.62 mg / g, the content of effective components is stable, and it meets the quality standard; The pills disintegrate within 30 minutes (meet the requirements of pills), and the fluctuation of the content of effective components is only 3.8% after 6 months of storage, meeting the requirements of preparation use.

[0060] Comparative Example 1 This comparative example provides the traditional YiShenJieBi Wan traditional hot reflux extraction-vacuum concentration process, which includes the following specific contents: Comparison purpose: Use the traditional hot reflux extraction and vacuum concentration process for comparison.

[0061] Comparison steps: (1) Preliminary processing of medicinal materials: As in Example 1, the medicinal materials are ground to 100 mesh.

[0062] (2) Hot reflux extraction: Put the fine powder of medicinal materials into the extraction tank, add solvent at a ratio of 1 part of medicinal materials to 12 parts of 50% alcohol solution, control the temperature to 75°C (traditional process temperature is high), and reflux extract for 2 hours (traditional process time is long), to obtain the extraction liquid.

[0063] (3) Filtration and residue removal: As in Example 1, filtration was performed using a 0.3 μm fine filter.

[0064] (4) Concentration under reduced pressure: The clear extract was placed in a concentration tank under reduced pressure, and the vacuum degree was controlled at -0.09 MPa and the temperature was controlled at 52°C, and the solution was concentrated to a relative density of 1.20 g / cm3 at 25°C.

[0065] Comparative effect: The dissolution ratio of the active ingredients was only 48.2%, which was 17.1 percentage points lower than that of Example 1, and did not meet the requirements; The alkaloid retention was 75.1% and the active peptide retention was 73.5%, which was 17-18 percentage points lower than that of Example 1, and the destruction of the heat-sensitive components was obvious; The unit energy consumption was 42% higher than that of Example 1, and the concentration time was 60% longer than that of Example 1, highlighting the advantages of the new method in terms of high efficiency, energy saving and quality preservation.

[0066] Compared with Example 1-5 and Comparative Example 1, Comparative Example 1 used the traditional production process of Yishen Juanbi Pills (preprocessing of medicinal materials → hot reflux extraction → filtration of the extract → concentration under reduced pressure), and the core parameters were a hot reflux temperature of 75°C, a reflux time of 2 h, and a concentration temperature under reduced pressure of 52°C. Examples 1-5 had the core process of "preprocessing - ultrasonic-assisted extraction - filtration of the extract - concentration by nanofiltration membrane", and only had special optimizations in the aspects of preprocessing, extraction mode, impurity removal, post-processing, etc.

[0067] From the core indicators, the active ingredient extraction rate of Comparative Example 1 was only 48.2%, and the alkaloid and active peptide retention rates were 75.1% and 73.5%, respectively, and the degradation of heat-sensitive components was obvious. Examples 1-5 were all better than this level: the active ingredient extraction rate of Example 1 (core process) was 65.3%, and the retention rate of heat-sensitive components was ≥91.8%; the extraction rate of Example 2 (with enzyme hydrolysis - ultrafine grinding) was increased to 68.7%, the extraction time of plant medicines was shortened by 20%, and the loss of active ingredients was only 2.2%; the extraction rate of insect active peptides of animal medicines in Example 3 (staged ultrasonic) was 57.2%, and the polysaccharide impurities of plant medicines were reduced by 13.5%; the removal rate of monosaccharides and inorganic salts in Example 4 (with resin impurity removal) was over 86%, and the recovery rate of alkaloids and active peptides was ≥95.7%; the moisture content of dry extract in Example 5 (with post-processing and formulation adaptation) was 4.2%, the disintegration time of the pills was 28 min, and the ingredient fluctuation was 3.8% after 6 months of storage.

[0068] From the process efficiency and energy consumption, the extraction time of Comparative Example 1 was 2 h, and the concentration time was 60% longer than that of Example 1, and the unit energy consumption was 42% higher than that of Example 1; the extraction time of Examples 1-5 was all controlled within 30-60 min, and the nanofiltration concentration temperature was 25-30°C, and the concentration time of Example 4 was further shortened by 15% due to the early impurity removal, and the overall energy consumption of Example 2 was significantly lower than that of the traditional process due to the reduction of extraction time caused by enzyme hydrolysis preprocessing.

[0069] In summary, the traditional process of Comparative Example 1 has the defects of low extraction rate, large loss of heat-sensitive components, high energy consumption and low efficiency; Examples 1-5 comprehensively surpass the traditional process in effective component extraction, heat-sensitive component retention, impurity control and energy consumption efficiency through new process and special optimization, and the subdivision optimization (enzymolysis, phased ultrasonic, resin impurity removal, preparation adaptation) of Examples 2-5 further meets the special needs of Yishen Juanbi Pills, i.e. slow dissolution of plant medicines, uneven extraction of animal medicines, low purity of concentrated liquid and the need for pill adaptation.

[0070] In addition, each function module in each embodiment of the present application can be integrated in one processing module, or each module can exist physically alone, or two or more modules can be integrated in one module.

[0071] The above is merely a specific implementation of the present application, but the protection scope of the present application is not limited thereto, any person skilled in the art can easily think of changes or replacements within the technical range disclosed in the present application, which should be included in the protection scope of the present application, therefore, the protection scope of the present application should be subject to the protection scope of the claims.

[0072] Finally: the above is merely a preferred embodiment of the present application, and is not used to limit the present application, any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application should be included in the protection scope of the present application.

Claims

1. A novel method for extracting and concentrating highly effective components of Yishen Juanbi Pill, characterized in that, Includes the following steps: Pre-treatment of medicinal materials: Take the medicinal materials prescribed for Yishen Juanbi Pill, which consist of Drynaria fortunei, Rehmannia glutinosa (processed), Angelica sinensis, Cynanchum paniculatum, Eupolyphaga sinensis, Scolopendra subspinipes, Buthus martensii, Vespa nidus (stir-fried), Pheretima aspergillum (processed with wine), Zaocys dhumnades (processed with wine), Corydalis yanhusuo, Pyrola rotundifolia, Epimedium brevicornu, Clematis chinensis, Geranium wilfordii, Spatholobus suberectus, Humulus scandens, Rehmannia glutinosa (raw), and Polygonum cuspidatum. The preparation conditions are as follows: add 5%-8% wheat bran by weight of the medicinal materials, and control the stir-frying temperature at 120-140°C. Stir-fry at 0℃ for 5-8 minutes; the preparation conditions for honeycomb (stir-fried) are: control the stir-frying temperature at 110-130℃ and stir-fry continuously for 3-5 minutes; the preparation conditions for earthworm (wine-processed) and black-striped snake (wine-processed) are: soak in 15%-20% yellow rice wine for 2-3 hours, and then dry in an environment of 50-60℃; wash and remove impurities from the above-mentioned medicinal materials separately, then mix and pulverize them to a particle size of 80-120 mesh to obtain medicinal powder; (2) Ultrasonic-assisted extraction: Add the medicinal powder obtained in step (1) to the extraction tank, add 30%-70% ethanol aqueous solution as the extraction solvent at a material-liquid ratio of 1:8-1:15, turn on the ultrasonic-assisted extraction device, control the ultrasonic power of 200-600W, the extraction temperature of 40-65℃, and the extraction time of 30-90min, and obtain the extract after the extraction is completed; (3) Filtration of the extract: The extract obtained in step (2) is filtered under reduced pressure using a 0.22-0.45μm microporous membrane to remove the filter residue and collect the clear extract. (4) Nanofiltration membrane concentration: The clarified extract obtained in step (3) is passed into a nanofiltration membrane separation system. The nanofiltration membrane is made of polyethersulfone or sulfonated polyethersulfone. The operating pressure is controlled at 1.0-3.0 MPa and the concentration temperature is controlled at 25-40℃ for low-temperature concentration treatment until the relative density of the extract at 25℃ reaches 1.10-1.30 g / cm³, and the concentrated solution of Yishen Juanbi Pill is obtained. In the method, the extraction rate of effective components from medicinal materials is ≥60%, and the retention rate of heat-sensitive active components is not less than 90%, wherein the heat-sensitive active components include alkaloids and active peptides.

2. The novel method for extracting and concentrating the highly efficient components of Yishen Juanbi Pill as described in claim 1, characterized in that, In step (1), the particle size of the pulverized medicinal material is 100 mesh, and the ambient temperature is controlled to not exceed 30℃ and the relative humidity is not exceed 60% during the pulverization process.

3. The novel method for extracting and concentrating the highly effective components of Yishen Juanbi Pill as described in claim 1, characterized in that, In step (2), the material-to-liquid ratio is 1:12, the volume fraction of the ethanol-water solution is 50%, the ultrasonic power is 400W, the extraction temperature is 55℃, and the extraction time is 60min.

4. The novel method for extracting and concentrating the highly effective components of Yishen Juanbi Pill as described in claim 1, characterized in that, In step (2), the ultrasound-assisted extraction process adopts an intermittent ultrasound mode, that is, the ultrasound works for 30 seconds and then pauses for 10 seconds, and the operation is repeated until the set extraction time is reached.

5. The novel method for extracting and concentrating the highly efficient components of Yishen Juanbi Pill as described in claim 1, characterized in that, In step (4), the molecular weight cutoff of the nanofiltration membrane is 100-500 Da, and the nanofiltration membrane is cleaned online every 30 minutes during the concentration process. The cleaning solution is a sodium hydroxide aqueous solution with a volume fraction of 0.5%-1.0%.

6. The novel method for extracting and concentrating the highly efficient components of Yishen Juanbi Pill as described in claim 1, characterized in that, It also includes step (5) post-treatment of the concentrate: the concentrate of Yishen Juanbi Pill obtained in step (4) is placed in a vacuum drying oven, the vacuum degree is controlled to be -0.08 to -0.10 MPa and the drying temperature is 50-60℃, and the moisture content is dried to ≤5% to obtain dry extract of Yishen Juanbi Pill, which is used to prepare pills, tablets or capsules.

7. The novel method for extracting and concentrating the highly efficient components of Yishen Juanbi Pill as described in claim 1, characterized in that, Step (1) of the pretreatment of medicinal materials also includes an enzymatic hydrolysis step: After cleaning and removing impurities, the following herbs are pulverized to 40-60 mesh: Drynaria fortunei, Rehmannia glutinosa (processed), Angelica sinensis, Cynanchum paniculatum, Corydalis yanhusuo, Pyrola rotundifolia, Epimedium brevicornu, Clematis chinensis, Geranium wilfordii, Spatholobus suberectus, Humulus scandens, Rehmannia glutinosa (raw), Polygonum cuspidatum, and Bombyx mori (stir-fried with wheat bran). A cellulase solution with a mass fraction of 0.1%-0.3% and an enzyme activity of 5000 U / g is added. The temperature is controlled at 45-55℃ and the pH at 4.5-5.

5. The process involves enzymatic hydrolysis for 1-2 hours. After enzymatic hydrolysis, the mixture is combined with pre-crushed materials such as ground beetle, centipede, scorpion, honeycomb (stir-fried), earthworm (processed with wine), and black-striped snake (processed with wine). The combined medicinal materials are then ultra-finely pulverized to 150-200 mesh to obtain medicinal powder. The enzymatic hydrolysis-ultra-fine pulverization process shortens the extraction time of effective components from plant-based medicinal materials such as Drynaria fortunei and Rehmannia glutinosa by 15%-25%, and the loss of effective components during pulverization is ≤3%.

8. The novel method for extracting and concentrating the highly efficient components of Yishen Juanbi Pill as described in claim 1, characterized in that, Step (2) Ultrasonic-assisted extraction is controlled in stages: In the first stage, only the powders of ground beetle, centipede, scorpion, honeycomb (stir-fried), earthworm (wine-processed), and black-striped snake (wine-processed) are ultrasonically extracted, with ultrasonic power controlled at 500-600W, temperature at 55-65℃, and time at 40-50min; In the second stage, the powders of Drynaria fortunei, Rehmannia glutinosa, Angelica sinensis, Cynanchum paniculatum, Corydalis yanhusuo, Pyrola rotundifolia, Epimedium brevicornu, Clematis chinensis, Geranium wilfordii, Spatholobus suberectus, Humulus scandens, Rehmannia glutinosa, and Polygonum cuspidatum are added to the extraction tank, and the ultrasonic power is adjusted to 300-400W, temperature at 40-50℃, and time at 20-40min to complete the extraction; The staged extraction results in an extraction rate of insect active peptides in animal-based medicinal materials such as ground beetle and centipede ≥55%, and a reduction of 10%-15% in the dissolution of polysaccharide small molecule impurities in plant-based medicinal materials such as Drynaria fortunei and Rehmannia glutinosa.

9. The novel method for extracting and concentrating the highly efficient components of Yishen Juanbi Pill as described in claim 1, characterized in that, Between steps (3) and (4), there is also step (3a) impurity adsorption: the clarified extract obtained in step (3) is passed into a resin column packed with D101 or AB-8 macroporous adsorption resin, the flow rate is controlled at 1-2 BV / h and the temperature at 25-30℃, after adsorption is completed, it is eluted with 10%-20% ethanol aqueous solution and the eluent is collected; the eluent is used as the feed liquid in step (4) for nanofiltration membrane concentration; the macroporous adsorption resin adsorption can remove 80%-90% of monosaccharides and inorganic salts in the extract, and the recovery rate of alkaloids and active peptides in Yishen Juanbi Pill is ≥95%.

10. The novel method for extracting and concentrating the highly efficient components of Yishen Juanbi Pill as described in claim 6, characterized in that, In step (5), the obtained dry extract of Yishen Juanbi Pill is pulverized to a particle size of 120-150 mesh to obtain dry extract powder. When the dry extract powder is used to prepare pills, it is mixed with excipients at a mass ratio of dry extract powder: excipients = 1:0.3-0.

8. The excipients include starch and microcrystalline cellulose.

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