CN106309783A - Rhizoma alismatis granules with efficacies of removing and preventing urinary calculus used for preventing and treating urinary calculus as well as preparation method and quality detection method - Google Patents
Rhizoma alismatis granules with efficacies of removing and preventing urinary calculus used for preventing and treating urinary calculus as well as preparation method and quality detection method Download PDFInfo
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Abstract
The invention provides rhizoma alismatis granules with the efficacies of removing and preventing urinary calculus used for preventing and treating urinary calculus. A preparation method of the rhizoma alismatis granules comprises the following steps: taking the five medicinal materials, including rhizoma alismatis, herba plantaginis, herba desmodii styracifolii, poria cocos and cassia twig as the raw materials, adding with 8-fold amount of water, decocting the medicinal materials for twice, wherein each time is 2h, mixing the decoctions, filtering the obtained mixed decoction, concentrating the filtrate to obtain extract, adding with 1.5-fold amount of dextrin and 0.004-fold amount of steviosin, carrying out uniform mixing by adopting 80% alcohol as a wetting agent (the use amount of alcohol is 10% of that of the extract), and preparing the granules by adopting a conventional method. A quality detection method is characterized in that a thin-layer chromatography of herba desmodii styracifolii and cassia twig in the medicinal materials is adopted as an identification method, specifically, the contents of two active ingredients, namely, plantamajoside and schaftoside in the medicinal materials are measured by adopting a high performance liquid chromatography. The product is used for preventing and treating urinary calculus, and an extraction process is simple, stable and feasible; a quality standard detection method of the rhizoma alismatis granules with the efficacies of removing and preventing urinary calculus is sensitive and reliable and is strong in specificity.
Description
Technical field
The invention belongs to field of medicaments, relate to medicine treating urinary system calculus and preparation method thereof and quality determining method.
Background technology
Urinary system calculus is a kind of common diseases of urinary system, also referred to as urinary calculus, including kidney, ureter, bladder and calculus of urethra, wherein most commonly seen with kidney and ureteral calculus.Having several factors to affect calculus, major sexual is other, age, race, heredity, dietary habit, environmental factors and occupation etc.[1].Incidence rate at mankind's urinary system calculus in life is about 10%-15%, and some middle east are even up to 20%-25%, and prevalence rises year by year[2].China's urinary calculi disease is the most common, is apt to occur in person between twenty and fifty, and sickness rate is gradually increasing in recent years.According to documents and materials, China's urinary system calculus sickness rate is 1%-5%, and southern area urinary system calculus sickness rate is more up to 5-10%[3].And, the feature of the high highly significant of urinary system calculus especially of relapse rate.It is reported, in the case of not taking any preventive measure, the urinary system calculus relapse rate of 1 year is close to 10%, and the relapse rate of 5 years is 33%, and the relapse rate of 10 years is 50%, and the relapse rate of 15 years is then up to 75%[4].So far there is no the recurrence of effective ways prevention urinary system calculus.
Calculus once generates, and often can cause urinary tract obstruction, secondary infection and epithelial lesion, and these are all that it produces the performance of secondary lesion to kidney.Do not discovered by patient when calculus is the least, have the calculus can discharged from health when proper motion.But, along with the increase of calculus volume, Most patients can the symptom such as, hematuria blunt with kidney district, also have some calculus can enter ureter and cause hydronephrosis or renal colic, make patient pain unbearably[5].Although additionally, stone is typically considered a kind of acute illness, but increasing evidence shows that calculus is that one can cause end stagerenaldisease[6], and can cause and suffer from hypertension[7], coronary heart disease[8], metabolism syndrome[9]And diabetes[10]Risk raise systemic function disorders.Therefore, urinary calculi to attract great attention, and not yet falls ill and their early stage in calculus, and calculus just should carry out preventing and treating when not yet becoming greatly in time.At present, extracorporeal shock-wave lithotripsy (ESWL) and intracavity urinary system technology can cure most of Patients with Urinary Stone by less invasive techniques, but, accepting the renal function of the patient of ESWL and Urology Surgery intracavity operation it may happen that damage, the relapse rate of urinary system calculus also can rise[11].Therefore, the medicine finding effectively prevention and treatment urinary system calculus is significant, is also the main research direction in this field and the important topic being jointly devoted to solution.
Summary of the invention
The task of the present invention is to provide Rhizoma Alismatis calculus anti-stone granule of a kind of prevention and treatment urinary system calculus and preparation method thereof.Another task of the present invention is to provide the quality determining method of described calculus anti-stone granule.
Realize the technical scheme is that prevention and the Rhizoma Alismatis calculus anti-stone granule for the treatment of urinary system calculus that the present invention provides, it is with Rhizoma Alismatis, Herba Plantaginis, Herba Desmodii Styracifolii, Poria and Ramulus Cinnamomi gomi herbs are raw material, the medicament made by the following method: take above-mentioned gomi herbs, add 8 times amount water, decoct secondary, each 2 hours, collecting decoction, filter, filtrate is concentrated at 80 DEG C the extractum obtaining relative density about 1.30, add 1.5 times of dextrin as filler, the steviosin of 0.004 times is correctives, with the ethanol of 80% concentration as wetting agent, ethanol consumption is 10% times of extractum amount, mixing, granule is made by well-established law, it is dried, granulate and get final product.The quality determining method of the calculus anti-stone granule that the present invention provides, is that the thin layer chromatography of Herba Desmodii Styracifolii and/or Ramulus Cinnamomi is as discrimination method in medicament, and specifically Sino-German to medicament Big Semen Plantaginis glycosides and/or two kinds of active constituent contents of Schaftoside are measured.Through zoopery and clinical research prove Rhizoma Alismatis calculus anti-stone granule that the present invention provides in animal body, In vitro and in vivo be respectively provided with the effect that significantly suppression calcium oxalate stone is formed.Dosage form selection of the present invention is granule, have an advantage in that: 1. Chinese medicine granules had not both had decoction problem in use, such as: the dosage taken of decoction is relatively big, decoction decocting inconvenient, decoction easily occur mould lose rotten, can have again some advantages, such as decoction absorbs fast, decoction effect rapidly etc.；2. the preparation technology of granule is suitable to commercial production；3. granule carrying, store, transport, the aspect such as to take the most convenient.Additionally, recipe quantity on the one of we is 54g, dried cream yield is about 10%, considers from dose, selects granule proper.Rhizoma Alismatis calculus of the present invention anti-stone granule is sugar type granules, reduces the ratio of adjuvant, makes dose be greatly decreased, and makes the grade of product be improved.It addition, in terms of Chinese medical theory, sugariness overrich, can grow and greasy hinder heresy, be unfavorable for the treatment of disease, the weak crowd of gastrointestinal function is the most uncomfortable.In terms of modern medicine, if modern study has turned out sugared part excess that urinary calculus patient takes in, it is unfavorable for the treatment of calculus, and the growth of promotion calculus can be made, make calculus become big.The Icing Sugar content of sugar-free granule is low, is more suitable for suffering from diabetes, hyperlipemia or old patient and takes.This project alternatives sugar-free granule, decrease patient's picked-up to sugar part, it is particularly suitable for urinary system calculus patient to use, solve the problems referred to above, expand clinical application range, create bigger economic and social benefit, make the medical value of this prescription obtain bigger performance and utilization, in like product, have more competitiveness simultaneously.
Experimental data
Study on extraction
Instrument: silica gel g thin-layer plate (Shanghai Ke Xing commerce and trade company limited)；Polyamide film (City of Taizhou road and bridge tetramethyl biochemistry plastic molding and processing plant product)；Glass point sample capillary tube (production of instrument plant of Huaxi Medical Univ)；Double-range electronic analytical balance AUW-220D type；Agilent 1100 series high performance liquid chromatograph；Shimadzu LC-20A type high performance liquid chromatograph；Agilent TC-C18 (4.6 × 250mm) chromatographic column；RE-52A type Rotary Evaporators (Ying Yu Yu Hua instrument plant of Gongyi City)；DZF-6020 type vacuum drying oven (Shanghai Suo Pu Instrument Ltd.)；Pulverizer (HY-02, converge rich sea of clouds development in science and technology company limited in Hebei)；HH-4 type digital display thermostat water bath (sunlight instrument plant of Jintan City of Jiangsu Province)；Electronic balance (JD1000-2, Longteng Electronic Weighing Instrument Co., Ltd., Shenyang)；Visible uv scan instrument (UV-1601PC type, Shimadzu International Trading Company Ltd).Reagent: acetonitrile (TEDIA company of the U.S., chromatographically pure), methanol (TEDIA company of the U.S., chromatographically pure), purified water；Methanol, toluene, ethanol, butanone, chloroform, n-butyl alcohol, ether, petroleum ether, acetic acid ethyl ester, hexamethylene, formic acid, glacial acetic acid, sulphuric acid, vanillin, aluminum chloride, silico-tungstic acid, dinitrophenylhydrazine (being analytical pure)；Neutral alumina (Qingdao Shuo Yuan Chemical Co., Ltd., chromatography is used)；(lot number is Alisol B monoacetate: 111846), (lot number is Big Semen Plantaginis glycosides: 111914), (lot number is cinnamic aldehyde: 110710), (lot number is Herba Desmodii Styracifolii control medicinal material: 121248), (lot number is Herba Plantaginis control medicinal material: 110710), (lot number is Poria control medicinal material: 121117), (lot number is Ramulus Cinnamomi control medicinal material: 121191), be all purchased from Nat'l Pharmaceutical & Biological Products Control Institute；(lot number is Schaftoside: 110227), is purchased from Chengdu Pu Feide biological product company limited.Medical material: (lot number is Rhizoma Alismatis: 130212), Herba Plantaginis: (lot number is: 130123), Herba Desmodii Styracifolii: (lot number is: 121209), (lot number is Poria: 130712), (lot number is Ramulus Cinnamomi: 130110), and above medical material is all purchased from Hubei Kang Sheng Pharmaceuticals Ltd.Surveyor: Tongji Medical College, Huazhong Science and Technology Univ. associate professor Yin Chunping.
Medical material medical material is checked: 1. quality of medicinal material standard: Rhizoma Alismatis is the tuber of version " Chinese Pharmacopoeia " Notes On Alism At Aceae Rhizoma Alismatis Alisma orientalis (Sam.) Juzep. recorded in 2010.The Rhizoma Alismatis bought should comply with pertinent regulations under version " Chinese Pharmacopoeia " Rhizoma Alismatis kind item in 2010.Herba Desmodii Styracifolii is the dry aerial parts of version " Chinese Pharmacopoeia " leguminous plant Herba Desmodii Styracifolii Desmodium styracifolium (Osb.) Merr. recorded in 2010[43].The Herba Desmodii Styracifolii medical material bought should comply with pertinent regulations under version " Chinese Pharmacopoeia " Herba Desmodii Styracifolii kind item in 2010.Herba Plantaginis is version " Chinese Pharmacopoeia " Plantaginaceae plant Herba Plantaginis Plantago asiatica L. recorded in 2010 or the dry herb of Plantago depressa Willd Plantago depressa Willd..The Herba Plantaginis medical material bought should comply with pertinent regulations under version " Chinese Pharmacopoeia " Herba Plantaginis kind item in 2010.Poria is the dry sclerotia of version " Chinese Pharmacopoeia " On Polyporaceae Poria Poria cocos (Schw.) Wolf recorded in 2010[43].The Poria medical material bought should comply with pertinent regulations under version " Chinese Pharmacopoeia " Poria kind item in 2010.Ramulus Cinnamomi is the dry twig of version " Chinese Pharmacopoeia " canella Cortex Cinnamomi Cinnamomum cassia Presl recorded in 2010[43].The Ramulus Cinnamomi medical material bought should comply with pertinent regulations under version " Chinese Pharmacopoeia " Ramulus Cinnamomi kind item in 2010.2. the qualitative identification of medical material: Rhizoma Alismatis carries out thin layer discriminating according to pertinent regulations under one Rhizoma Alismatis Variety identification item of " Chinese Pharmacopoeia " version in 2010.Rhizoma Alismatis test sample is aobvious same color speckle on relevant position with Alisol B monoacetate and Rhizoma Alismatis control medicinal material, meets regulation.Herba Plantaginis carries out thin layer discriminating according to pertinent regulations under one Herba Plantaginis Variety identification item of " Chinese Pharmacopoeia " version in 2010.Under uviol lamp, Herba Plantaginis test sample is aobvious same color fluorescence speckle on relevant position with Herba Plantaginis control medicinal material and Herba Plantaginis reference substance Big Semen Plantaginis glycosides, meets regulation.Herba Desmodii Styracifolii carries out thin layer discriminating according to pertinent regulations under one Herba Desmodii Styracifolii Variety identification item of " Chinese Pharmacopoeia " version in 2010.Under uviol lamp, Herba Desmodii Styracifolii test sample is aobvious same color fluorescence speckle on relevant position with Herba Desmodii Styracifolii reference substance Schaftoside and Herba Desmodii Styracifolii control medicinal material, meets regulation.Poria carries out thin layer discriminating according to pertinent regulations under one Poria Variety identification item of " Chinese Pharmacopoeia " version in 2010.Poria test sample and Poria control medicinal material be aobvious same color speckle on relevant position, meets regulation.Ramulus Cinnamomi carries out thin layer discriminating according to pertinent regulations under one Ramulus Cinnamomi Variety identification item of " Chinese Pharmacopoeia " version in 2010.Ramulus Cinnamomi test sample is aobvious same color speckle on relevant position with Ramulus Cinnamomi reference substance cinnamic aldehyde and Ramulus Cinnamomi control medicinal material, meets regulation.
2.2.2.3 the assay of medical material: 1. Rhizoma Alismatis carries out assay according to pertinent regulations under one Rhizoma Alismatis kind assay item of " Chinese Pharmacopoeia " version in 2010.Recording the content of Alisol B monoacetate in Rhizoma Alismatis is 0.059%, more than 0.050%, meets regulation.2. Herba Plantaginis carries out assay according to pertinent regulations under one Herba Plantaginis kind assay item of " Chinese Pharmacopoeia " version in 2010.Recording the content of Big Semen Plantaginis glycosides in Herba Plantaginis medical material is 0.72%, more than 0.10%, meets regulation.3. Herba Desmodii Styracifolii carries out assay according to pertinent regulations under one Herba Desmodii Styracifolii kind assay item of " Chinese Pharmacopoeia " version in 2010.Recording the content of Schaftoside in Herba Desmodii Styracifolii medical material is 0.14%, more than 0.13%, meets regulation.4. without assay requirement under one Poria kind item of Poria " Chinese Pharmacopoeia " version in 2010.5. Ramulus Cinnamomi carries out assay according to pertinent regulations under one Ramulus Cinnamomi kind assay item of " Chinese Pharmacopoeia " version in 2010.In the Ramulus Cinnamomi medical material recorded, Determination of Cinnamaldehyde is 0.10%, equal to 0.10%, meets regulation.
2.2.3 medical material water absorption rate is investigated with soak time: Chinese medicinal material extraction process is to improve the basis of extractum quality, extraction process must improve quality and yield, shorten man-hour with the extractum of preparation on the premise of ensureing curative effect as far as possible, reduce cost, reduce material consumption energy consumption.Physicochemical property design extraction process according to effective ingredient or effective site, with water boiling and extraction.For ensureing that we have stable curative effect, improve the extracted amount of effective ingredient, should first soak, owing to soak time is less with other factor reciprocal actions, it be carried out single factor exploration.Extraction is made a strong impact by soak time with water absorption rate.But because this examination can need not survey active constituent content with visual evaluation, exclude in formal screening programme.
2.2.3.1 experimental technique: the most accurately weigh one day recipe quantity (54g) Chinese crude drug, totally 9 parts, it is placed in 500ml beaker, numbering, then it is separately added into water 300ml, start to soak, the soak time of 9 parts of medical materials is followed successively by 10,20,30,40,50,60,70,80,90 (min), then filter to the greatest extent, medicinal residues are weighed, finally calculates water absorption rate.Computing formula: water absorption rate (%)=(medicinal residues weight in wet base-medical material dry weight)/medical material dry weight × 100%.
2.2.3.2 experimental result: result is as follows, is shown in Table 1, Fig. 1.
Table 1 medical material water absorption rate and soak time investigation table
2.2.3.3 interpretation of result: medical material is after soaking 40min, and water absorption basically reaches balance, illustrates that the water absorption of medical material had been maxed out value when 40 minutes, and water absorption now is the 179.63% of medical material total amount.From producing reality, in order to allow medical material fully soak, being beneficial to extract, the final soak time selecting medical material is 40min.
2.2.4 orthogonal test research is extracted
2.2.4.1 quadrature factor water-glass: application " orthogonal design assistant 3.1 " software, it is three inspection target with Schaftoside, Big Semen Plantaginis glycosides extracted amount and dried cream yield, with extraction time, amount of water, extraction time for investigating factor, carrying out orthogonal test, its factor level is shown in Table 2.
Table 2 water extraction factor level table
2.2.4.2 test method: after determining that soak time is 40min, in addition it is also necessary to influence factor's impacts on the extraction ratio of medical material such as examination extraction time, extraction time, amount of water.Therefore design orthogonal test, for index, it is investigated with dried cream yield, extracts active ingredients amount, screen optimum extraction process.Having gomi herbs in we's extraction process by water, according to composition and the dosage form requirement of five kinds of Chinese medicine, selecting dried cream yield, Big Semen Plantaginis glycosides, Schaftoside three is evaluation index, carries out overall merit.Dried cream yield: a is as the reference index of selection dosage form, if it is the highest, it is difficult to make granule；B affects the selection of impurity-removing method, dry degree, the stability of finished product；C is granule day dosing and the foundation that determines of packing specification.Therefore must be using dried cream yield as one of evaluation index, but it does not become dose-effect relationship with curative effect intensity, therefore during overall merit, weight coefficient should be less, the most suitable with 0.1.Big Semen Plantaginis glycosides, Schaftoside are one of effective ingredient of Herba Plantaginis, the index components of Herba Desmodii Styracifolii and the anti-stone of calculus respectively, and weight coefficient should be relatively large, is respectively advisable with 0.45.Factor level table according to above-mentioned design, select L9 (34) orthogonal design table experiment arrangement: take 0.2 times of recipe quantity medical material, 720g altogether, carry out testing (when adding water for the first time by the requirement of orthogonal design table, 1296ml water is added) by medical material water absorption rate 180%, it is concentrated into 500ml (1.44g crude drug/ml) after filtrate being merged, standby.Content assaying method further accordance with Big Semen Plantaginis glycosides, Schaftoside measures Big Semen Plantaginis glycosides and the Schaftoside content in concentrated solution, and calculates the extracted amount of 0.2 times of recipe quantity medical material；Measure dried cream yield.
2.2.4.2.1 the content assaying method (seeing 3.2.7) of Big Semen Plantaginis glycosides: 1. chromatographic condition and system suitability test: with acetonitrile-water (18:82) for flowing phase；With 18 alkyl silica gel as filler；UV-detector.Number of theoretical plate is calculated by Big Semen Plantaginis glycosides and is not less than 3000.2. the preparation of reference substance solution: it is appropriate that precision weighs Big Semen Plantaginis glycosides reference substance, record, the methanol solution adding 60% makes every 1ml solution containing about 15 μ g, calculates, to obtain final product.The preparation of 3 need testing solutions: precision measures orthogonal water boiling and extraction liquid 1ml, place it in evaporating dish, water bath method, residue dissolves with the methanol of 60% and is transferred in the volumetric flask of 10ml, the methanol solution adding 60% is to volumetric flask scale, shaking up, microporous filter membrane (0.45 μm) filters, and to obtain final product.4 algoscopys: accurate Big Semen Plantaginis glycosides reference substance solution, each 10 μ l of need testing solution of drawing, in injection high performance liquid chromatograph, measure, then calculate the content of Big Semen Plantaginis glycosides, to obtain final product.
2.2.4.2.2 the method (seeing 3.2.7) of Schaftoside assay: 1. chromatographic condition and system suitability test: with acetonitrile-water (16:84) for flowing phase；With 18 alkyl silica gel as filler；UV-detector.Number of theoretical plate is calculated by Schaftoside and is not less than 3000.2. the preparation of reference substance solution: it is appropriate that precision weighs Schaftoside reference substance, record adds 50% methanol and makes every 1ml solution containing about 30 μ g, calculates, to obtain final product.3. the preparation of need testing solution: precision measures orthogonal water boiling and extraction liquid 1ml, place it in evaporating dish, water bath method, residue dissolves with the methanol of 50% and is transferred in the volumetric flask of 10ml, the methanol solution adding 50% again is to volumetric flask scale, shaking up, microporous filter membrane (0.45 μm) filters, and to obtain final product.4. algoscopy: accurate Schaftoside reference substance solution, each 10 μ l of need testing solution of drawing, in injection high performance liquid chromatograph, is measured, then calculates the content of Schaftoside, to obtain final product.
2.2.4.2.3 dried cream yield assay method: precision measures each group of orthogonal each 50ml of soak by water concentrated solution respectively, it is respectively placed in drying again in the clean evaporating dish of constant weight, by its water bath method, then, place in 105 DEG C of dry 3h in baking oven, take out, be placed in exsiccator placement 30min, weigh, calculate dried cream yield.
2.2.4.3 result of the test: result of the test is shown in Table 3, table 4.
Table 3. extraction process by water Orthogonal Experiment and Design and result
Note: dried cream yield scoring=(10/ maximum dried cream yield) × dried cream yield
Big Semen Plantaginis glycosides scoring=(45/ maximum Big Semen Plantaginis glycosides amount) × Big Semen Plantaginis glycosides amount
Schaftoside scoring=(45/ maximum Schaftoside amount) × Schaftoside amount
Comprehensive grading=dried cream yield scoring+Big Semen Plantaginis glycosides scoring+Schaftoside scoring
Table 4 variance analysis
| Soruces of variation | Deviation square | Degree of freedom | Variance | F ratio | Significance |
| A | 119.20 | 2 | 59.60 | 2.21 | |
| B | 208.42 | 2 | 104.21 | 3.86 | |
| C | 2579.20 | 2 | 1289.60 | 47.82 | * |
| D | 53.95 | 2 | 26.97 |
F0.05 (2,2)=19, * significance
Intuitively being analyzed from upper table, the practice factor order affecting extraction effect is: extraction time > extraction time > amount of water, wherein with extraction time impact maximum.Understanding through above-mentioned variance analysis the most again, extraction time, amount of water are on extraction effect there are no significant impact, and extraction time has a significant impact, and optimal extraction time is 2h, considers extraction efficiency and cost.The soak by water optimised process finally determined is: A1B2C3, it may be assumed that 8 times amount water, to extract 2 times, each extraction time is 2h.
2.2.4.4 extraction process checking: weigh medical material, totally three parts, by A1B2C3Carry out checking test, operation repetitive, the results are shown in Table 5.
The checking test of table 5 extraction process by water
Result shows, this process stabilizing, feasible.
2.3 separation, purification, concentration technology research
2.3.1 separate, purifying process is studied: Chinese medicinal material has the macromole impurity of a lot of silt, medicinal residues fragment and invalid thing in the decocting liquid after decocting, these materials are the principal elements affecting Chinese medicine preparation dose and mouthfeel.Appointed condition according to industrialized production and technological requirement, select Filtration and sedimentation to process.Filtration: use rustless steel flame filter press.Medicinal liquid can be removed most solid impurity from extraction pot by plate filter filtration.Sedimentation: the medicinal liquid after plate filter filters is stored in vertical settling tank and stands overnight, has many precipitations in the bottom of settling tank, the fragment of precipitation mainly medical material and insoluble impurity.
In industrial operation, it is contemplated that practical situation, determine remove impurity in the following order:
2.3.2 concentrate and study with drying process: concentration condition: the medicinal liquid after filtration, settlement treatment, use concentrating under reduced pressure tank, be 1.30 (80 DEG C) by concentration of liquid medicine to relative density.
Drying condition: the dry employing of granule 60 DEG C～80 DEG C is dried.
2.4. preparations shaping technical study: Chinese medical concrete mixes with appropriate excipient, after soft material processed, sieves, and is dried, just can be prepared as granule after granulate.Application comparison Chinese medicine preparation excipient widely has sucrose, starch, dextrin, lactose etc..It is reported, urinary system calculus patient eats sugar too much, not only hinders calculi therapy, also can promote calculus further growth [44,45], therefore the adjuvant of this product can not use sucrose.And, this granule is water-soluble granular formulation, therefore can not select starch is excipient.Chinese medicine sugar-free granule, the most the more commonly used excipient has dextrin, soluble starch, lactose, microcrystalline Cellulose etc..Add different types of excipient, the quality of Chinese medicinal granule can be produced a very large impact.This experiment, according to relevant documents and materials and trial test result, compares analysis to dextrin, soluble starch and lactose etc..With ratio of briquetting, moisture, melting as inspection target, carry out comprehensive evaluation.
2.4.1 instrument and reagent
2.4.1.1 instrument: sieve (No., No. four, pharmacopeia is sieved)；Electric heating constant-temperature blowing drying box (DHG-914, the permanent Science and Technology Ltd. in Shanghai one).
2.4.1.2 reagent: soluble starch (Tianjin Tian Li chemical reagent company limited)；Dextrin, mannitol, microcrystalline Cellulose (MCC) (Beijing extensive and profound in meaning star biotechnology responsibility company limited)；Lactose (Shanghai reagent three factory)；Ethanol (Tianjin chemical reagent three factory)；Steviosin (Qufu Xiang Zhou Flos Chrysanthemi goods Co., Ltd).
2.4.2 adjuvant screening
2.4.2.1 minimum supplementary product consumption is investigated
2.4.2.1.1 experimental technique: weigh the extractum l0g under 2.3.2 item, totally 12 parts, mix in the ratio in table 6 with soluble starch, dextrin, lactose, mannitol, microcrystalline Cellulose respectively, ethanol solution with 85% is as wetting agent, to " hands is pinched agglomerating, pressure the most scattered ", make soft material, granule is made again with the sieve of No., it is positioned in time in baking oven, is dried 5 hours in 80 DEG C, granule is taken out, finally with the sieve granulate of No. four, to obtain final product.
2.4.2.1.2 experimental result: the results are shown in Table 6.
Table 6 adjuvant minimum amount investigation table
2.4.2.1.3 interpretation of result: can draw to draw a conclusion from upper table 6, extractum is 1:1.5 with adjuvant soluble starch, the minimum amount ratio of dextrin；Extractum is 1:3 with adjuvant lactose, the minimum scale of mannitol；Extractum is 1:1 with the minimum scale of microcrystalline Cellulose.
The investigation of the most single adjuvant
2.4.2.2.1 test method: weigh the extractum 10g under 2.3.2 item, totally 5 parts, respectively with adjuvant soluble starch, dextrin, lactose, mannitol, microcrystalline Cellulose mixing, the consumption of adjuvant is 30g, ethanol solution with 85% is as wetting agent, make soft material, become granule with a sieve series, granule is placed in baking oven, 80 DEG C are dried 5 hours, taking-up cools, then with the sieve granulate of 12 mesh, to obtain final product.Measure the most using the following method prepared the ratio of briquetting of five parts of particulate samples, melting, hygroscopicity.The mensuration of ratio of briquetting: the Rhizoma Alismatis calculus anti-stone granule prepared is weighed, record, the most first to cross a sieve, collect the granule by a sieve, this partial particulate, after No. four sieves, is collected and can not be weighed by the granule of No. four sieves, record.Each group of grain forming rate is calculated again respectively by equation below.
Granular mass × 100% before granular mass after ratio of briquetting=sieve/sieve.
The mensuration of melting: according to " Chinese Pharmacopoeia " 2010 editions one annex IC granule " melting " defined, takes the granule 10g prepared, and adds hot water 200ml, stirs to all, records the time, observes.
Hygroscopic mensuration: weigh a certain amount of Rhizoma Alismatis calculus anti-stone particulate samples, be placed in constant weight 48 hours in the baking oven of 30 DEG C.In bottom is placed with the glass desicator of NaCl saturated solution, NaCl is put in timing, until forming the supersaturated solution of NaCl, the relative humidity being now placed with in the glass desicator of NaCl saturated solution is 75%.Putting into the thick granule of about 2mm bottom the flat weighing botle of constant weight, correct amount is placed in above-mentioned exsiccator (flat weighing botle is opened) and again weighs after 48 hours, calculates Moisture percentage, respectively does two groups, calculate its meansigma methods.The hydroscopicity of each group of granule is calculated again respectively by equation below.
Weight × 100% before Moisture percentage %=(weight before weight-moisture absorption after moisture absorption)/moisture absorption
2.4.2.2.2 result of the test: the results are shown in Table 7.
Table 7 particulates' properties index determining table
| Adjuvant | Ratio of briquetting (%) | Solution time (S) | 72h hydroscopicity (%) |
| Dextrin | 91.40 | 125 | 19.16 |
| Lactose | 92.30 | 80 | 12.74 |
| Mannitol | 91.60 | 90 | 11.96 |
| Soluble starch | 87.20 | 150 (in suspendible shapes) | 19.86 |
| Mcc | 92.10 | There is precipitation | 14.35 |
Ratio of briquetting: lactose > MCC > mannitol > dextrin > soluble starch.
Thawing time: soluble starch > dextrin > mannitol > lactose.
Hydroscopicity: soluble starch > dextrin > MCC > lactose > mannitol.
2.4.2.2.3 interpretation of result: comprehensive every result, MCC is water insoluble, unavailable；Lactose, mannitol ratio of briquetting are high, solution time is short, hydroscopicity is little, but supplementary product consumption is relatively big, expensive, the most inapplicable；Dextrin, soluble starch minimum supplementary product consumption are less, it may be considered that.
2.4.2.3 the investigation of mixed accessories
2.4.2.3.1 test method: weigh the extractum 10g under 2.3.2 item, totally 6 parts, mixing with the mixed accessories of different proportion respectively by table 8, the consumption of mixed accessories is 30g, ethanol solution with 85% is as wetting agent, make soft material, become granule with a sieve series, granule is placed in baking oven, 80 DEG C are dried 5 hours, taking-up cools, then with a sieve granulate, to obtain final product.Measure respectively by the method in 2.4.2.2.1 prepared the ratio of briquetting of six parts of particulate samples, melting, hygroscopicity.
2.4.2.3.2 result of the test: the results are shown in Table 8.
Table 8 different proportion mixed accessories is preferred
| Adjuvant | Ratio | Solution time (s) | Ratio of briquetting (%) |
| Dextrin: soluble starch | 5:0 | 120 | 95.26 |
| 4:1 | 130 | 93.38 | |
| 3:2 | 130 | 93.26 | |
| 2:3 | 130 | 93.35 | |
| 1:4 | 140 | 89.83 | |
| 0:5 | 150 | 87.22 |
2.4.2.3.3 interpretation of result: the effect individually pelletized with dextrin is substantially better than dextrin soluble starch mixed accessories mixing granulation；Finally select with dextrin as diluent；With measuring its minimum supplementary product consumption 1:1.5.
2.4.2.4 the selection of correctives and the screening of consumption: Rhizoma Alismatis calculus anti-stone granular raw medicinal liquid is more bitter, oral is difficult to swallow, and needs to improve its mouthfeel.According to Rhizoma Alismatis calculus anti-stone granule contained drug composition and the feature of dosage form, select to add correctives the most suitable.Sweeting agent should be added for covering the bitterness of medicinal liquid.Steviosin is also referred to as stevioside, is leaf of Folium Stevlae Rebaudianae seed extract, without sugar and heat；Color and luster white is to the most slightly yellow, and mouthfeel is suitable, free from extraneous odour, its natural low caloric value and closely sucrose taste, and sugariness is about 150～300 times of sucrose, is the wide new sugar source of development prospect.Steviosin belongs to non-fermented material, stable in properties, is difficult to go mouldy, and is eaten for a long time and will not cause dental caries, advantageously reduces viscosity, bacteria growing inhibiting simultaneously, extends shelf life of products.Steviosin is the sweeting agent that the current world has been found that and uses through the approval of China Ministry of Public Health, is to continue outside Caulis Sacchari sinensis, beet sugar that the third has Development volue and healthy natural sweetener praised highly, is described as " third place in the world sugar source " in the world.In a word, steviosin have cheap, by the advantage such as thermally-stabilised, high safety, the high and low heat of sugariness, pure in mouth feel, be preferable sweeting agent.Therefore select steviosin to replace sucrose as sweeting agent.
2.4.2.4.1 test method: take the extractum 30g under 2.3.2 item, adds the dextrin (45g) of 1.5 times, and the ethanol with 85%, as wetting agent, makes granule.Take prepared granule 30g, be equally divided into 6 parts, every part of 5g, add steviosin, mixing in table 9 ratio, be separately added into after 100ml boiled water dissolves, many people taste.
2.4.2.4.2 result of the test: the results are shown in Table 9.
Table 9 steviosin consumption optimization test result
| Sampling amount (g) | Steviosin usage ratio (%) | Mouthfeel |
| 5 | 0.1 | Bitter |
| 5 | 0.2 | Bitter |
| 5 | 0.3 | Slightly bitter |
| 5 | 0.4 | Moderate |
| 5 | 0.5 | Cross sweet |
| 5 | 0.6 | Cross sweet |
2.4.2.4.3 interpretation of result: as can be known from the above table, steviosin consumption is when 0.4%, and mouthfeel is moderate, the most properly.
2.4.3 pelletizing orthogonal test research: in industrialized production, most widely used a kind of method of granulating is exactly wet granulation at present, it of main cause is the highest to equipment requirements, easy and simple to handle.Therefore Rhizoma Alismatis calculus anti-stone granule also selects wet granulation.Extractum i.e. adds 1.5 times of dextrin and the steviosin of 0.004 times, makes granule, be dried.And screen optimal granulating process condition with orthogonal experiment method.
2.4.3.1 test method: from preliminary experiment and documents and materials, extractum relative density, the consumption of wetting agent ethanol and concentration are the principal element affecting grain forming rate, respectively take three levels, carries out orthogonal test, and gauge outfit design is shown in Table 10.
Table 10 factor level table
According to above-mentioned factor level table, select L9(34) orthogonal design table tests, it may be assumed that take 100g clear paste, add the dextrin of 1.5 times, the steviosin of 0.4% times, test by the orthogonal table requirement of design.The another ratio of briquetting calculating granule respectively.
2.4.3.2 result of the test: result of the test is such as table 11 below, as shown in 12.
Table 11. Orthogonal Experiment and Design and result
Table 12 variance analysis
| Soruces of variation | Sum of square of deviations | Degree of freedom | F ratio | Significance |
| A | 684.14 | 2 | 19 | * |
| B | 71.28 | 2 | 19 | |
| C | 79.20 | 2 | 19 | |
| D | 33.95 | 2 | 19 |
" * " represents significance.
2.4.3.3 interpretation of result: process table 11, table 12 data and variance analysis, with the percentage rate of conforming particle as evaluation index, result shows that factor A has significance: A2>A3>A1, optimum A should be taken2, factor B, C do not have significance；The selected optimum process condition of experiment is combined as A2B2C2, i.e. extractum relative density is 1.30, sprays into 10% times amount 80% alcohol granulation.
2.4.3.4 verification experimental verification: verify 3 times by the most selected experimental technique, calculate the ratio of briquetting of granule respectively, the results are shown in Table 13.
Result of the test verified by table 13
Result above shows, the checking result of the test of Rhizoma Alismatis calculus anti-stone granule formation technology is consistent with orthogonal experiments, and this process stabilizing, feasible is described.Dose processing method of the present invention and process chart are shown in Fig. 2 and embodiment.
The quality standard of 3.2 Rhizoma Alismatis calculus anti-stone granule is formulated and drafts explanation
3.2.1 title: Rhizoma Alismatis calculus anti-stone granule: we are made up of Rhizoma Alismatis, Herba Plantaginis, Herba Desmodii Styracifolii, Poria and Ramulus Cinnamomi five kinds of Chinese medicine.Can be clinically used for functioning of bladder unfavorable, the damp-heat accumulation part of the body cavity below the umbilicus, housing the bladder, kidneys and bowels, urine is decocted by damp and hot, and turbidity condenses, and hinders mechanism of qi, the not smooth caused sand Stranguria of QI-blood circulation, stranguria caused by urinary stone.In side, " diuretic the first non-defective unit " Rhizoma Alismatis is monarch drug, and prescription has effect of clearing away heat-damp and promoting diuresis, Tonglin Paishi, therefore named " the anti-stone of Rhizoma Alismatis calculus ".Granule has conveniently, stable, in good taste, can pour in water drink into, apply and carry more convenient, dissolution and the advantage such as infiltration rate is very fast, therefore dosage form selection is granule.According to the relevant regulations " name of drug product; before nomenclature of drug arranges; after dosage form ranks " in " China's adopted drug name " nomenclature principle, " before in drug product title, the adjective of illustrative purposes or feature etc. is preferably listed in medicine name ", " use main Chinese medicinal materials name and function to combine and add dosage form name ", by named for this product " Rhizoma Alismatis calculus anti-stone granule "
3.2.2 prescription: same to text, sees above.
3.2.3 preparation method: same to text, sees above.
3.2.4 the same text of character.Each batch of finished product of pilot scale all describes consistent with text.
3.2.5 differentiate
3.2.5.1 instrument and reagent: silica gel g thin-layer plate (Shanghai Ke Xing commerce and trade company limited), polyamide film (City of Taizhou road and bridge tetramethyl biochemistry plastic molding and processing plant product), glass point sample capillary tube (production of instrument plant of Huaxi Medical Univ), HH-4 type digital display thermostat water bath (sunlight instrument plant of Jintan City of Jiangsu Province), ultrasonic washing unit (MP5200H, Shaanxi Peng Zhan Science and Technology Ltd.), ten very much one electronic analytical balances (Shenyang Longteng Electronic Co., Ltd.)；
Methanol, water, acetic acid, butanone, formic acid, chloroform, ether, petroleum ether, ethyl acetate, sulphuric acid, vanillin, aluminum chloride；(lot number is Schaftoside reference substance: 110227) be purchased from Chengdu Pu Feide biological product company limited；(for differentiating, lot number is Herba Desmodii Styracifolii control medicinal material: 121248), (for differentiating, lot number is Ramulus Cinnamomi control medicinal material: 121191), be purchased from Nat'l Pharmaceutical & Biological Products Control Institute；
Rhizoma Alismatis calculus anti-stone granule (20130912,20130916,20130920, self-control).
3.2.5.2 the thin layer of Herba Desmodii Styracifolii differentiates: the preparation of need testing solution takes Rhizoma Alismatis calculus anti-stone particulate samples 1g, finely ground, puts in tool plug conical flask, add the methanol 25ml of 80%, supersound process 20min, filter, filtrate is evaporated, and the methanol solution of residue 10ml50% dissolves, both.3 batch samples are prepared with method.The preparation of control medicinal material solution: take Herba Desmodii Styracifolii control medicinal material powder 0.2g, prepare with method with test sample.The preparation of reference substance solution: take Herba Desmodii Styracifolii reference substance Schaftoside appropriate, adds 50% methanol and prepares the Schaftoside reference substance solution of about 75ug/ml.The preparation of negative control solution: weigh the ingredients in addition to Herba Desmodii Styracifolii according to recipe quantity, negative granules sample is prepared by the preparation method of Rhizoma Alismatis calculus anti-stone granule, take 1g again, prepare according to the preparation method of need testing solution and lack Herba Desmodii Styracifolii negative control solution.
Thin layer chromatography: take three batches of need testing solutions, reference substance solution, negative sample solution, each 1ul of control medicinal material solution, put respectively on same polyamide film, with methanol-water-acetic acid (10:10:0.5) solution as developing solvent, launch, taking out, dry, spray is with 1%AlCl3Developer, dries up, and puts and inspects under uviol lamp (365nm).In test sample chromatograph, with on reference substance chromatograph, the corresponding position of control medicinal material chromatograph, show identical weak blue fluorescence speckle.Negative control solution is without corresponding speckle, noiseless, sees Fig. 3.In the quality standard text of income this product.
3.2.5.3 the thin layer of Ramulus Cinnamomi differentiates: 1, the preparation of need testing solution takes Rhizoma Alismatis calculus anti-stone particulate samples 5g, finely ground, is placed in tool plug conical flask, adds ether 30ml, soaking 20min, shake constantly, filter, filtrate is evaporated, residue adds chloroform 1ml, makes dissolving, to obtain final product.3 batch samples are prepared with method.2, the preparation of control medicinal material solution takes Ramulus Cinnamomi control medicinal material powder 5g, add 200ml soak by water 30min, filtering, medicinal residues add 200ml soak by water 30min again, merge decoction liquor and are concentrated into 20ml, extract 2 times with ether 20ml shaking, combined ether layer, water bath method, residue adds chloroform 1ml, make dissolving, to obtain final product.3, the preparation of negative sample solution takes recipe quantity ingredients in addition to Ramulus Cinnamomi and prepares negative sample by the preparation method of finished product, then takes 5g, lacks Ramulus Cinnamomi negative sample solution by the preparation method preparing need testing solution with method preparation.4, thin layer is extracted need testing solution, negative sample solution, each 10ul of control medicinal material solution, put respectively on same silica gel g thin-layer plate, with petroleum ether (60-90 DEG C)-ethyl acetate (15:5) solution as developing solvent, launch, take out, drying, spray is with 2% vanillin-sulfuric acid solution developer, and 105 DEG C are heated to colour developing uniformly.In test sample chromatograph, on position corresponding with control medicinal material chromatograph, aobvious identical yellow principal spot.Negative control solution then without corresponding speckle, does not interferes with and sees Fig. 4.In the quality standard text of income this product.
3.2.5.4 Rhizoma Alismatis thin layer differentiate: use various method, in test sample chromatograph, with on reference substance chromatograph, the corresponding position of control medicinal material chromatograph, all without corresponding speckle.Therefore do not take in the quality standard text of this product.
3.2.5.5 Herba Plantaginis thin layer differentiate: use various method, in test sample chromatograph, with on reference substance chromatograph, the corresponding position of control medicinal material chromatograph, all without corresponding speckle.Therefore do not take in the quality standard text of this product.
3.2.5.6 the thin layer of Poria differentiates: use various method, all cannot get rid of negative interference.Therefore do not take in the quality standard text of this product.
3.2.6 check
3.2.6.1 granularity: according to the assay method of defined in " granularity " under version " Chinese Pharmacopoeia " annex IC granule item in 2010: use the granularity of double sieve method stone anti-to Rhizoma Alismatis calculus granule to check, take test sample 30g (taking unit dose package granule 6 to wrap), its weight weighed, it is placed in a sieve, transfers on No. four sieves.Left and right comes and goes, and keeps level to sieve, and sieves while tapping 3 minutes, takes and can not cross a sieve and can be calculated the percentage ratio of granule gross weight shared by it by No. four granules sieved and powder, weighed gross weight, must not be crossed 15% [43].The results are shown in Table 14.
Table 14 granularity checks result table
Result shows, the granularity of Rhizoma Alismatis calculus anti-stone granule meets regulation.
3.2.6.2 moisture: according to the assay method of defined in " moisture " under version " Chinese Pharmacopoeia " annex IC granule item in 2010: take granule 2-5g, accurately weighed, it is laid in drying in the flat weighing botle of constant weight, its thickness is less than 5mm, accurately weighed, open lid, 105 DEG C of baking ovens are dried 5 hours, bottle cap is built, move in exsiccator, after cooling down 0.5 hour, accurately weighed, it is dried 1 hour under the conditions of said temperature again, to constant weight, the weight alleviated according to granule, calculate the water content (%) of granule, 6.0% [43] must not be crossed.The results are shown in Table 15.
Result table is examined in table 15 moisture inspection
| Lot number | 1 | 2 | 3 | Mean (%) | RSD (%) |
| 130912 | 4.37 | 4.34 | 4.51 | 4.41 | 2.06 |
| 130916 | 3.95 | 3.95 | 4.08 | 3.99 | 1.88 |
| 130920 | 4.18 | 4.12 | 4.26 | 4.19 | 1.68 |
Result shows, the moisture of Rhizoma Alismatis calculus anti-stone granule meets regulation.
3.2.6.3 melting: according to the assay method of defined in " Chinese Pharmacopoeia " 2010 editions one annex IC granule " melting ": take test sample granule one bag, add hot water 200ml, stirs five minutes, observes [43] immediately；The results are shown in Table 16.
Table 16 melting checks result table
| Lot number | Melting |
| 130912 | All dissolve |
| 130916 | All dissolve |
| 130920 | All dissolve |
Result shows, the melting of Rhizoma Alismatis calculus anti-stone granule meets regulation.
3.2.6.4 content uniformity: according to the assay method [43] of defined in " Chinese Pharmacopoeia " 2010 editions one annex IC granule " content uniformity ": take test sample 10 bags, the respectively weight of weighed every bag of content, every packed amount is compared with indicating loading amount.The content uniformity of stone granule anti-to Rhizoma Alismatis calculus checks, the results are shown in Table 17.
Table 17 content uniformity checks result table
| Lot number | Weight differential |
| 130912 | Meet regulation |
| 130916 | Meet regulation |
| 130920 | Meet regulation |
Result shows, the content uniformity of Rhizoma Alismatis calculus anti-stone granule meets regulation.
3.2.6.5 microbial limit: according to the detection method [43] of defined in " Chinese Pharmacopoeia " 2010 editions one annex IC granule " microbial limit ": the number of antibacterial should≤1000/g；Mycete, saccharomycetic number≤100/g；Escherichia coli must not detect.Check with conventional method stone anti-to Rhizoma Alismatis calculus granule.The results are shown in Table 18.
Table 18 limit test of microbe
Result shows, the microbial limit of Rhizoma Alismatis calculus anti-stone granule meets regulation.
In sum, the relevant check item of Rhizoma Alismatis calculus anti-stone granule all meets regulation, can perform above-mentioned standard.
3.2.7 assay
3.2.7.1 instrument and reagent: 100,000/electronic analytical balance (Shenyang Longteng Electronic Co., Ltd.), double-range electronic analytical balance AUW-220D type (SHIMADZU CORPORATION JAPAN), Agilent 1100series high performance liquid chromatograph, Agilent TC-C18 (4.6mm × 250mm) chromatographic column；
Acetonitrile (TEDIA company of the U.S., chromatographically pure), redistilled water；
(lot number is Big Semen Plantaginis glycosides reference substance: 111914), for containing surveying, being purchased from Nat'l Pharmaceutical & Biological Products Control Institute；(lot number is Schaftoside reference substance: 110227) for containing surveying, being purchased from Chengdu Pu Feide biological product company limited.
3.2.7.2 the foundation of Big Semen Plantaginis glycosides content assaying method
3.2.7.2.1 chromatographic condition: determine that UV detection wavelength is 330nm, sample size is 10 μ l, by changing mobile phase ratio, and flow velocity etc., using target peak retention time T, peak type, separating degree and symmetry etc. as inspection target, selecting optimum chromatogram condition, the chromatographic condition finally determined is: chromatographic column: Agilent TC-C18 (4.6mm × 250mm) chromatographic column；Flowing phase: acetonitrile-water (18:82)；Flow velocity: 0.7ml/min；UV detects wavelength: 330nm；Column temperature: 30 DEG C；Sample size: 10 μ l.Under selected conditions, Big Semen Plantaginis glycosides and other component color spectral peaks can reach baseline separation, and Big Semen Plantaginis glycosides chromatographic peak is more than 1.5 with adjacent chromatographic peak separating degree；Theoretical cam curve is calculated more than 3000 by Big Semen Plantaginis glycosides.
3.2.7.2.2 the preparation of reference substance solution: it is appropriate that precision weighs Big Semen Plantaginis glycosides reference substance, adds 60% methanol and makes reference substance solution (0.0212mg/ml), 0.45um membrane filtration, takes subsequent filtrate, to obtain final product.
3.2.7.2.3 the preparation of need testing solution: take this product under content uniformity item, mixing, take appropriate, finely ground, take about 2.5g, accurately weighed, being placed in tool plug conical flask, precision measures 50ml 60% methanol solution, weighs, ultrasonic (power 250W, frequency 50KHZ) 30min, lets cool, weigh, supply the weight of less loss with 60% methanol solution, with 0.45 μm membrane filtration, take subsequent filtrate, to obtain final product.
3.2.7.2.4 specificity test: preparation lacks Herba Plantaginis negative granules sample, is prepared by the compound method of need testing solution.Drawing reference substance solution, need testing solution, each 10 μ l of negative sample solution respectively, inject in chromatograph of liquid and measure, result shows: Herba Plantaginis negative sample solution does not interferes with.See Fig. 5, Fig. 6, Fig. 7.
3.2.7.2.5 the investigation of linear relationship: precision pipettes 0.1ml, 0.2ml, 0.4ml, 0.6ml, 0.8ml, 1.0ml reference substance solution in 1ml volumetric flask respectively, and 60% methanol constant volume is the reference substance of series concentration.Measuring the peak area of each group of concentration Big Semen Plantaginis glycosides reference substance under fixed chromatographic condition, with sample size (C) as abscissa, peak area (A) is vertical coordinate, Criterion curve.Trying to achieve regression equation is: y=35456x 18.539, r=0.9992；Result show Big Semen Plantaginis glycosides sample size in 0.0212～0.212 μ g linearly (be shown in Table 19, Fig. 8).
Result investigated by table 19 linear relationship
3.2.7.2.6 precision test: accurate absorption Big Semen Plantaginis glycosides reference substance solution, repeats sample introduction 6 times under the chromatographic condition determined, records peak area, seek RSD value.
Table 20 Big Semen Plantaginis glycosides precision test (n=6)
3.2.7.2.7 stability test: take finite concentration Big Semen Plantaginis glycosides reference substance solution, under the conditions of HPLC in 0,1,2,4,6,8h sample introduction respectively, record peak area, seek RSD value.Shown in result see table.
Table 21 Big Semen Plantaginis glycosides stability test
3.2.7.2.8 replica test: by test sample processing method, precision weighs same batch sample 6 parts, measures each group of peak area by the chromatographic condition determined, calculates content and RSD value result is as shown in the table
Table 22 Big Semen Plantaginis glycosides replica test (n=6)
3.2.7.2.9 accuracy test (recovery test): precision weighs 0.25g this product powder in 6 10ml tool plug conical flasks respectively, the Schaftoside solution 1.5ml drawing 0.212mg/ml respectively adds, another addition 8ml 60% methanol solution, separately take 1.5ml Schaftoside solution, be not added with sample, be set to blank group, after processing constant volume by test sample preparation method, measure each group of peak area by the HPLC method set up, calculate the content of Big Semen Plantaginis glycosides, calculate the response rate.Because to have measured the average content of Big Semen Plantaginis glycosides in sample in replica test be 0.1296%, then adding the known quantity (mg) of sample=weigh powder quality (mg) × 0.1296%, its result see table.
Table 23 Big Semen Plantaginis glycosides response rate result (n=6)
Recovery test result shows: the Big Semen Plantaginis glycosides response rate is between 95%～100%, and sample-adding reclaims good.
3.2.7.2.10 the determination of content limit: stone granule ten batch sample anti-to Rhizoma Alismatis calculus is measured, and the results are shown in Table 24.
Big Semen Plantaginis glycosides assay result in table 24 sample
According to said determination result, considering actual content and the factor such as preparation technology, big production of Herba Plantaginis medical material, tentative this product every bag (every packed 5g granule), with Big Semen Plantaginis glycosides (C29H36O16) meter, 5.0mg must not be less than.
3.2.7.3 the foundation of Schaftoside content assaying method
3.2.7.3.1 chromatographic condition: determine that UV detection wavelength is 272nm, sample size is 10 μ l, by changing mobile phase ratio, and flow velocity etc., using target peak retention time T, peak type, separating degree and symmetry etc. as inspection target, selecting optimum chromatogram condition, the chromatographic condition finally determined is: chromatographic column: Agilent TC-C18 (4.6mm × 250mm) chromatographic column；Flowing phase: acetonitrile-water (16:84)；Flow velocity: 0.8ml/min；UV detects wavelength: 272nm；Sample size: 10 μ l.Under selected conditions, Schaftoside and other component color spectral peaks can reach baseline separation, and Schaftoside chromatographic peak is more than 1.5 with adjacent chromatographic peak separating degree；Theoretical cam curve is pressed Schaftoside and is calculated more than 3000.
3.2.7.3.2 the preparation of reference substance solution: it is appropriate that precision weighs Schaftoside reference substance, adds 50% methanol and makes reference substance solution (0.01288mg/ml), 0.45 μm membrane filtration, takes subsequent filtrate, to obtain final product.
3.2.7.3.3 the preparation of need testing solution: take this product under content uniformity item, mixing, take appropriate, finely ground, take about 1g, accurately weighed, being placed in tool plug conical flask, precision measures 50ml 50% methanol solution, weighs, ultrasonic (power 250W, frequency 50KHZ) 30min, lets cool, weigh, supply the weight of less loss with 50% methanol solution, with 0.45 μm membrane filtration, take subsequent filtrate, to obtain final product.
3.2.7.3.4 specificity test: preparation lacks Herba Desmodii Styracifolii negative granules sample, prepares according to the compound method of need testing solution.Drawing reference substance solution, need testing solution, each 10 μ l of negative sample solution respectively, inject in chromatograph of liquid and be measured, result shows: negative sample solution does not produce interference (see Fig. 9, Figure 10, Figure 11).
3.2.7.3.5 linear relationship is investigated: take Schaftoside reference substance solution (0.01288mg/ml), precision pipettes 0.1ml, 0.2ml, 0.4ml, 0.6ml, 0.8ml, 1.0ml reference substance solution in 1ml volumetric flask respectively, and 50% methanol constant volume is the reference substance of series concentration.Measuring the peak area of each group of concentrations control product under fixed chromatographic condition, with the concentration of each group of solution Schaftoside as abscissa, peak area A is vertical coordinate, Criterion curve.Trying to achieve regression equation is: y=37299X-16.1114, r=0.9995.Result shows that Schaftoside sample size linearly (is shown in Table 25, Figure 12) in 0.01288～0.1288 μ g.
Result investigated by table 25 linear relationship
3.2.7.3.6 precision is investigated precision and is drawn Schaftoside reference substance solution, repeats sample introduction 6 times, record peak area, seek RSD value under the chromatographic condition determined.
Table 26 Schaftoside precision test (n=6)
3.2.7.3.7 stability test: take finite concentration Schaftoside reference substance solution, under the conditions of HPLC in 0,1,2,4,6,8h sample introduction respectively, record peak area, seek RSD value.Shown in result see table.
Table 27 Schaftoside stability test
3.2.7.3.8 replica test: by test sample processing method, precision weighs same batch sample 6 parts, measures each group of peak area by the chromatographic condition determined, calculates content and RSD value result is as shown in the table
Table 28 Schaftoside replica test (n=6)
3.2.7.3.9 accuracy test (response rate): precision weighs 0.1g this product powder in 6 10ml tool plug conical flasks respectively, pipette, extract 0.1288mg/ml Schaftoside reference substance solution 2ml, another addition 7ml 50% methanol solution, separately take 2ml reference substance solution, be not added with sample, be set to blank group, after processing constant volume by test sample preparation method, measure each group of peak area by the HPLC method set up, calculate the content of Schaftoside, calculate the response rate.Because to have measured Schaftoside content in sample in replica test be 0.2581%, then adding the known quantity (mg) of sample=weigh powder quality (mg) × 0.2581%, its result see table.
Table 29 Schaftoside response rate result (n=6)
3.2.7.3.10 the determination of content limit: ten batch samples of stone granule anti-to Rhizoma Alismatis calculus carry out assay, mensuration the results are shown in Table 30.
Schaftoside assay table in table 30 sample
According to said determination result, considering actual content and the factor such as preparation technology, big production of Herba Desmodii Styracifolii medical material, tentative this product every bag (every packed 5g) is with Schaftoside (C26H28O14) calculate, 8.0mg must not be less than.
3.2.7.4 brief summary: establish the thin-layer identification method of Herba Desmodii Styracifolii, Ramulus Cinnamomi in the quality standard of Rhizoma Alismatis calculus anti-stone granule；Establish the high performance liquid chromatography content assaying method of Big Semen Plantaginis glycosides, Schaftoside.Empirical tests, methodological science, stable, reliable.
Accompanying drawing explanation
Fig. 1 medical material water absorption rate investigates figure.
The process chart of Fig. 2 Rhizoma Alismatis calculus anti-stone granule.
Fig. 3 thin-layer chromatogram.
Fig. 4 thin-layer chromatogram.
Fig. 5 Big Semen Plantaginis glycosides reference substance HPLC schemes.
Fig. 6 Rhizoma Alismatis calculus anti-stone particulate samples HPLC schemes.
Fig. 7 negative sample Solution H PLC is schemed.
Fig. 8 Big Semen Plantaginis glycosides standard curve.
Fig. 9 Schaftoside reference substance HPLC schemes.
Figure 10 Rhizoma Alismatis calculus anti-stone particulate samples HPLC schemes.
Figure 11 negative sample Solution H PLC is schemed.
Figure 12 Schaftoside standard curve.
Detailed description of the invention
Embodiment 1: take Rhizoma Alismatis, Herba Plantaginis, Herba Desmodii Styracifolii, Poria and Ramulus Cinnamomi gomi herbs, adds 8 times amount water, decocts secondary, each 2 hours, collecting decoction, filter, filtrate is concentrated into appropriate relative density about 1.30 (80 DEG C), add 80% ethanol of 1.5 times of dextrin, the steviosin of 0.004 times and 10% times amount, mixing, make granule, it is dried, granulate, makes 1000g, to obtain final product.The process chart of the Rhizoma Alismatis calculus anti-stone granule that the present invention provides is shown in Fig. 2.The function of the Rhizoma Alismatis calculus anti-stone granule that the present invention provides cures mainly: clearing away heat and promoting diuresis, the anti-stone of calculus.Curing mainly the pyretic stranguria caused by damp-heat in lower-JIAO, stranguria caused by urinary stone, disease sees yellowish urine dysurea, waist abdomen pain；Lithangiuria (renal calculus, ureteral calculus), is shown in above-mentioned disease person.The formation of prevention urinary system calculus and recurrence.The usage and dosage of suggestion: boiled water is taken after mixing it with water.One time 1 bag, 3 times on the one.Suggestion specification: every packed 5g.Sealing storage, effect duration fixes tentatively 12 months.Process chart is shown in Fig. 2.The medical material of one recipe quantity (54g) of this granule can go out cream about 6g, calculates with daily three times, and need to take dry cream amount is 2g every time.Medicine is 1:1.5 with the ratio of adjuvant.Therefore this product particle size is about every packed 5g.Calculating by 1000 preparation unit i.e. (l000g), one preparation prescription crude drug amount of Rhizoma Alismatis calculus anti-stone granule is 3600g, makes granule l000g.
Claims (5)
1. a prevention and the Rhizoma Alismatis calculus anti-stone granule for the treatment of urinary system calculus, it is characterised in that it is with Rhizoma Alismatis, car
Front grass, Herba Desmodii Styracifolii, Poria and Ramulus Cinnamomi gomi herbs are raw material, the medicament made by the following method: take above-mentioned gomi herbs,
Adding 8 times amount water, decoct secondary, each 2 hours, collecting decoction, filtration, filtrate was concentrated at 80 DEG C to obtain relative density about 1.30
Extractum, adding 1.5 times of dextrin as the steviosin of filler, 0.004 times is correctives, with the ethanol of 80% concentration as wetting agent,
Ethanol consumption is 10% times of extractum amount, mixing, makes granule by well-established law, is dried, granulate and get final product.
2. the quality determining method of the anti-stone of calculus described in claim 1 granule, it is characterised in that with Herba Desmodii Styracifolii in medicament and/
Or the thin layer chromatography of Ramulus Cinnamomi is as discrimination method.
3. the quality determining method of the anti-stone of calculus described in claim 1 granule, it is characterised in that to the Big Semen Plantaginis glycosides in medicament
And/or two kinds of active constituent contents of Schaftoside are measured.
The quality determining method of calculus the most according to claim 3 anti-stone granule, it is characterised in that use efficient liquid phase
Big Semen Plantaginis glycosides in medicament and/or two kinds of active constituent contents of Schaftoside are measured by chromatograph.
5. a preparation method for the Rhizoma Alismatis calculus anti-stone granule of prevention and treatment urinary system calculus, comprises the following steps: take
Rhizoma Alismatis, Herba Plantaginis, Herba Desmodii Styracifolii, Poria and Ramulus Cinnamomi gomi herbs are raw material, add 8 times amount water, decoct secondary, each 2 hours,
Collecting decoction, filters, and filtrate is concentrated at 80 DEG C to obtain the extractum of relative density about 1.30, add 1.5 times of dextrin as filler,
The steviosin of 0.004 times is correctives, and with the ethanol of 80% concentration as wetting agent, ethanol consumption is 10% times of extractum amount, and mixing is pressed
Well-established law makes granule, is dried, granulate and get final product.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109557234A (en) * | 2019-01-29 | 2019-04-02 | 广西壮族自治区药用植物园 | A kind of herba lysimachiae drenches clearly the thin-layer chromatography detection method of particle |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101953883A (en) * | 2010-10-22 | 2011-01-26 | 重庆科瑞制药有限责任公司 | Chinese medicinal granules for treating calculosis and preparation method thereof |
| CN102028827A (en) * | 2010-11-22 | 2011-04-27 | 姜永芳 | Medicinal composition for treating urinary tract infection and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101953883A (en) * | 2010-10-22 | 2011-01-26 | 重庆科瑞制药有限责任公司 | Chinese medicinal granules for treating calculosis and preparation method thereof |
| CN102028827A (en) * | 2010-11-22 | 2011-04-27 | 姜永芳 | Medicinal composition for treating urinary tract infection and preparation method thereof |
Non-Patent Citations (1)
| Title |
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| 杨明主编: "《中药药剂学》", 31 July 2012, 中国中医药出版社 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109557234A (en) * | 2019-01-29 | 2019-04-02 | 广西壮族自治区药用植物园 | A kind of herba lysimachiae drenches clearly the thin-layer chromatography detection method of particle |
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