CN102353675A - Identification method of Armillaria tabescens capsule medicament - Google Patents
Identification method of Armillaria tabescens capsule medicament Download PDFInfo
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Abstract
The invention discloses an identification method of an Armillaria tabescens capsule medicament, which is prepared from Armillaria tabescens leavening, and contains no less than 60 mg of polysaccharide in per gram calculated by C6H12O6 and no less than 60 mg of polypeptide calculated by bovine serum albumin. The identification method of the Armillaria tabescens capsule medicament comprises steps of: adding 40-60 ml of distilled water into 1g of the capsule content, dissolving by heating and filtering with absorbent cotton; adding 0.5 ml of ninhydrin test solution into 1.8-2.3ml of filtrate and heating; if the filtrate turns to ianthinus, the content contains Armillaria tabescens medicament. According to the identification method of Armillaria tabescens capsule medicament, simple identification and determination of the medicament can be conveniently carried out, without quantitative analysis; therefore the method is especially suitable for primary hospitals.
Description
The application is " a kind of Armilariella tabescens pharmaceutical capsules and discrimination method and quality standard detecting method ", the applying date to be on 06 28th, 2006 the dividing an application of No. 200610021284.8 patented claims for title.The notification of examiner's opinion first time of this original bill application points out that the original bill application does not have unicity, and based on this kind reason, the applicant proposes the application of this division.For fear of identical with original bill application title and can't distinguish, and for the better content of reflection this case application, the applicant is decided to be the title of this division application " a kind of discrimination method of Armilariella tabescens pharmaceutical capsules ".
Technical field
The present invention relates to a kind of Armilariella tabescens pharmaceutical capsules and discrimination method and quality standard detecting method.
Background technology
Armilariella tabescens (Armillarella tabescens (Scop.exFr) Sing) is a kind of ability enhances human body physique; Improve immunologic function; And have anti-inflammatory, pain relieving, fall enzyme, the medicinal fungi of removing jaundice and choleretic effect, its preparation is clinical to be usually used in treating leukopenia that acute and chronic hepatitis, metastatic hepatitis, chronic cholangitis and cholecystitis and chronic, shallow table property, atrophic gastritis, tympanitis and radiotherapy chemotherapy cause etc.But existing preparation is liquid preparation, and does not have capsule preparations, and the discriminating and the detection difficult of the Armilariella tabescens extract medicine of solid, and poor repeatability makes that the Armilariella tabescens pharmaceutical capsules of solid can't suitability for industrialized production and clinical practice.The Armilariella tabescens medicine of liquid preparation, the problem of its existence is: the active drug component content is low, and curative effect has much room for improvement, and its shelf-life is short, and storage transports and takes equal inconvenience, the clinical use that serious harm has hindered the Armilariella tabescens medicine.
Summary of the invention
The object of the present invention is to provide a kind of Armilariella tabescens pharmaceutical capsules, the active drug component content of this kind Armilariella tabescens pharmaceutical capsules is high, good effect, and long shelf-life, storage transport and take all convenient.
The objective of the invention is to realize through following technical scheme:
A kind of Armilariella tabescens pharmaceutical capsules is made by the Armilariella tabescens fermentate, it is characterized in that: the polyoses content in every gram content of pharmaceutical capsules is with anhydrous dextrose (C
6H
12O
6) meter>=60mg, content of peptides is in bovine serum albumin(BSA)>=60mg.
Compared with prior art, the invention has the beneficial effects as follows: drug ingedient is present in the capsule with the solid mode, and storage is transported, carries and take all very convenient, and its medicine active drug component content is high, not perishable, long shelf-life.
The agent property of above-mentioned medicine is a hard capsule, and content is sundown or buff powder, gas delicate fragrance.
Another object of the present invention provides a kind of quality standard detecting method of above-mentioned Armilariella tabescens pharmaceutical capsules; This mass standard detecting method precision is high; Favorable reproducibility, reliable and stable, can fine control product quality to guarantee the product quality and the clinical efficacy of suitability for industrialized production.
The quality standard detecting method of a kind of Armilariella tabescens pharmaceutical capsules provided by the invention is divided into two contents of mensuration of polyoses content and content of peptides, is characterized in that described measurement of the polysaccharide content step is:
Learn from else's experience 102 ℃-110 ℃ anhydrous dextrose reference substance that is dried to constant weight of the preparation of A, reference substance solution adds water and processes the solution that every 1ml contains anhydrous dextrose 0.08-0.12mg.
The preparation a of B, need testing solution, get the content of Armilariella tabescens pharmaceutical capsules, porphyrize, accurately claim decide about 1g and write down the amount of taking by weighing; Put in the measuring bottle, add water 35-45ml, put in the 75-85 ℃ of water bath with thermostatic control and heated 25-35 minute, jolting is constantly taken out, and is chilled to room temperature, and thin up shakes up to 50ml; B, filter with absorbent cotton, discard initial filtrating, precision is measured 1ml filtrating, puts in the 10ml centrifuge tube, adds absolute ethyl alcohol 6.5-7.5ml; C, on vortex suspendible device, mix 25-35 second, leave heart 12-18 minute with per minute 2500-3000; D, get deposition and add absolute ethyl alcohol 5.5-6.5ml, repeat the c step in step, get deposition again and add absolute ethyl alcohol 5.5-6.5ml, repeat the c step in step; E, the deposition water that d step is obtained go in the measuring bottle, and thin up shakes up to 100ml, as need testing solution.
The preparation precision of C, typical curve is measured reference substance solution 0,0.2,0.4,0.8,1.0ml; Put respectively in the tool plug test tube; Respectively add water to 1.0ml, add 3% phenol solution 0.8-1.3ml again, shake up; Add sulfuric acid 4.0-5.0ml rapidly; Shaking up, put and be chilled to room temperature, is blank with 0 pipe of not measuring reference substance solution; Measure absorbance log according to spectrophotometric method in the wavelength of 490nm, the regression equation of the amount that calculates glucose and corresponding absorbance log.
D, mensuration precision are measured need testing solution 1.0ml; Add 3% phenol solution 0.8-1.2ml again; Shake up; Add sulfuric acid 4.5ml rapidly, shake up, put and be chilled to room temperature; With 0 pipe of C in the step is blank; Measure absorbance log according to spectrophotometric method in the wavelength of 490nm,, go out the polyoses content of test sample again by the regression equation calculation in C step according to the absorbance log that records; According to the medicine amount of taking by weighing of B step record, calculate the polyoses content of every gram capsule 's content again.
The quality standard detecting method of a kind of Armilariella tabescens pharmaceutical capsules of the present invention, wherein the assay method of content of peptides is:
The bovine serum albumin(BSA) reference substance is got in the preparation of A, reference substance solution, adds water and processes the solution that every 1ml contains 0.25-0.35mg.
The content of Armilariella tabescens pharmaceutical capsules is got in the preparation of B, need testing solution, and porphyrize, the accurate title decide about 1g, and the record amount of taking by weighing; Place measuring bottle, add water 35-45ml, put in the 75-85 ℃ of water bath with thermostatic control and heated 25-35 minute, jolting is constantly taken out, and is chilled to room temperature, and thin up shakes up to 50ml; Filter with absorbent cotton, discard initial filtrating, precision is measured 2ml filtrating, places measuring bottle, and thin up shakes up to 100ml.
The preparation of C, forint phenol test solution:
The preparation of forint phenol first liquid: a, get sodium potassium tartrate tetrahydrate 0.4-0.6g, add water 50ml dissolving, b, get copper sulphate 0.4-0.6g; Add water 60ml dissolving; C, get NaOH 8-12g and sodium carbonate 45-55g, add water 400ml dissolving, face with preceding a, b, three kinds of solution of c got respectively by 1: 1: 8 mixing.
The preparation of forint phenol test solution second liquid: with 8 times of above-mentioned forint phenol first liquid thin ups.
The preparation precision of D, typical curve is measured reference substance solution 0,0.2,0.4,0.8,1.0ml; Put respectively in the tool plug test tube; Respectively add water to 1.0ml, add forint phenol test solution first liquid 1.0ml again, shake up; Room temperature was placed after 8-12 minute; Add forint phenol test solution second liquid 4.0ml, shake up, insulation is 4-7 minute in 50-60 ℃ of water-bath; Take out, put and be chilled to room temperature; 0 pipe not measure reference substance solution is blank, according to spectrophotometric method in the wavelength of 650nm mensuration absorbance log, and the regression equation of the amount that calculates bovine serum albumin(BSA) and corresponding absorbance log.
E, mensuration precision are measured need testing solution 1.0ml, add forint phenol test solution first liquid 1.0ml again, shake up, and room temperature was placed after 8-12 minute, add forint phenol test solution second liquid 4.0ml, shake up, and insulation is 4-7 minute in 50-60 ℃ of water-bath, takes out, and puts and is chilled to room temperature; With 0 pipe of D in the step is blank, measures absorbance log according to spectrophotometric method in the wavelength of 650nm, is gone out the content of peptides of need testing solution again by the regression equation calculation in D step; According to the medicine amount of taking by weighing of B step record, calculate the content of peptides of every gram capsule 's content again.
Following experiment showed, above-mentioned quality standard detecting method of the present invention, precision meets the requirements, favorable reproducibility, and sample was measured stable content in 4 hours.This quality standard detecting method can fine control product quality.
1, supplies the stability test of reagent matter sample: adopt quality standard detecting method of the present invention to 5 parts in a collection of Armilariella tabescens pharmaceutical capsules sample of the present invention; Process the need testing solution that polysaccharide and polypeptide are measured; Every interval was measured its polyoses content and content of peptides in 60 minutes, result such as following table:
| Minute | 0 minute | 60 minutes | 120 minutes | 180 minutes | 240 minutes | RSD |
| Polyoses content | 78.6 | 78.4 | 78.5 | 78.4 | 78.7 | 0.17％ |
| Content of peptides | 72.3 | 72.4 | 72.1 | 72.2 | 72.1 | 0.18％ |
Polysaccharide RSD=0.17% polypeptide RSD=0.18%
Show that need testing solution is stable at 4 hours intensive amounts.
2, precision test: get 8 parts of Armilariella tabescens capsules, 5 every part, measure its polysaccharide and peptide content respectively, result such as following table:
| Sample number | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | RSD |
| Polyoses content | 78.5 | 78.6 | 78.3 | 78.4 | 78.7 | 78.5 | 78.4 | 78.5 | 0.16％ |
| Content of peptides | 72.1 | 72.4 | 72.3 | 72.2 | 72.1 | 72.3 | 72.2 | 72.3 | 0.92％ |
Polysaccharide RSD=0.16% (N=8) polypeptide RSD=0.92% (N=8)
Show that quality standard detecting method precision of the present invention meets the requirements.
3, reappearance test: get with 6 parts of a collection of drug samples, 5 every part, measure its polysaccharide and peptide content respectively, result such as following table:
| Sample number | 1 | 2 | 3 | 4 | 5 | 6 | RSD |
| Polyoses content | 78.6 | 78.5 | 78.4 | 78.7 | 78.3 | 78.5 | 0.18％ |
| Content of peptides | 72.3 | 72.2 | 72.4 | 72.1 | 72.3 | 72.2 | 0.15％ |
Polysaccharide RSD=0.18% (N=6) polypeptide RSD=0.15% (N=6)
Show quality standard detecting method favorable reproducibility of the present invention.
4, the practice examining of middle trial production three batch products quality situation
To Armilariella tabescens pharmaceutical capsules of the present invention, three batch products of trial production are checked in getting according to quality standard detecting method of the present invention, and result's requirement all up to specification sees the following form.
It is thus clear that; Through above-mentioned quality standard detecting method; The polysaccharide of the Armilariella tabescens pharmaceutical capsules of the present invention that can guarantee to industrially produce and the content of two kinds of active drug compositions of polypeptide reach required standard; Guarantee pharmaceutical capsules component content consistance; Thereby guarantee the curative effect of medicine, in order to its extensive clinical use.
The 3rd purpose of the present invention provides the discrimination method of above-mentioned Armilariella tabescens pharmaceutical capsules:, under the situation that does not need quantitative test, this medicine is simply differentiated and judged with convenient, especially be fit to basic hospital and use.
First kind of discrimination method of above-mentioned Armilariella tabescens pharmaceutical capsules is: get capsule 's content 1g, and adding distil water 40-60ml heating for dissolving, absorbent cotton filters; Get filtrating 1.8-2.3ml, add ninhydrin solution 0.5ml, after the heating, be bluish violet and judge that then content contains the Armilariella tabescens medicine.
The step of second kind of discrimination method of above-mentioned Armilariella tabescens pharmaceutical capsules is:
A, get the capsule 's content porphyrize, take by weighing 0.9-1.1g, put in the measuring bottle, add water 35ml-45ml, put in 75 ℃ of-85 ℃ of waters bath with thermostatic control heating 25-35 minute, jolting is constantly taken out, and puts and is chilled to room temperature, and thin up shakes up to 50ml.
B, the solution that A is gone on foot filter with absorbent cotton, discard initial filtrating, and precision is measured 1ml filtrating, puts in the 10ml centrifuge tube, adds absolute ethyl alcohol 6.5-7.5ml, on vortex suspendible device, mix 25-35 second, leave heart 12-18 minute with per minute 2500-3000.
C, the deposition of getting B step adds water 2ml makes dissolving, moves in the test tube, adds 15% methyl naphthol ethanolic solution 1-3 and drips, and test tube wall adds the 0.7-1.5ml concentrated sulphuric acid, and two liquid meet the place, boundary if become purplish red colour circle, judge that then content contains the Armilariella tabescens medicine.
The step of the third discrimination method of above-mentioned Armilariella tabescens pharmaceutical capsules is:
A, get capsule 's content 1.5g, add water 40-60ml, boiled 12-18 minute, filter with absorbent cotton while hot; Filtrating is put cold, and the 18-23ml jolting that adds diethyl ether is extracted, and obtains ether solution, filters with absorbent cotton; Filtrating volatilizes, and residue adds absolute ethyl alcohol 0.5ml dissolving, as need testing solution.
B, get the armillarisin A reference substance in addition, add absolute ethyl alcohol and process the solution that every 1ml contains 8-12ug, as reference substance solution.
C, according to according to thin-layered chromatography test; Draw above-mentioned A, each 8-12u l of two kinds of solution in B step, put respectively on same silica gel g thin-layer plate, be 8: 2: 0.5 with proportioning: the upper strata liquid of normal butyl alcohol-methyl alcohol of 10-strong ammonia solution-water is developping agent; Launch; Take out, dry, under ultraviolet lamp (365nm), inspect; In the test sample chromatogram; With the corresponding position of reference substance chromatogram on, be the fluorescence spot of same color, then judge to supply to contain the Armilariella tabescens medicine in the examination capsule 's content.
More than three kinds of discrimination methods all can be under the situation that does not need complicated instrument; Can differentiate pharmaceutical capsules of the present invention; Judge whether capsule 's content contains the Armilariella tabescens medicine; Thereby conveniently medicine of the present invention is differentiated simply basic hospital that condition is relatively poor and pharmacy also can differentiate according to this.Thereby the clinical expansion that helps medicine of the present invention more uses.
Below in conjunction with embodiment the present invention is done further detailed description.
Embodiment one
A kind of Armilariella tabescens pharmaceutical capsules by the fermentate that Armilariella tabescens obtains, is promptly processed this medicine through existing extract drugs technology extraction and capsule preparations technology after fermentation.Through checking: the polyoses content in every gram content of pharmaceutical capsules is with anhydrous dextrose (C
6H
12O
6) meter>=60mg, content of peptides is specification product in bovine serum albumin(BSA)>=60mg.Formulation generally adopts hard capsule, and its content is sundown or buff powder, gas delicate fragrance.Certainly formulation also can be selected soft capsule for use.
The quality standard method of inspection of the pharmaceutical capsules that this is routine is divided into two contents of mensuration of polyoses content and content of peptides.
Wherein measurement of the polysaccharide content step is:
The learn from else's experience 105 ℃ anhydrous dextrose reference substance that is dried to constant weight of the preparation of A, reference substance solution adds water and processes the solution that every 1ml contains anhydrous dextrose 0.1mg.
The preparation a of B, need testing solution, get the content of Armilariella tabescens pharmaceutical capsules, porphyrize, accurately claim decide about 1g and write down the amount of taking by weighing; Put in the measuring bottle, add water 40ml, put in 80 ℃ of waters bath with thermostatic control and heated 30 minutes, jolting is constantly taken out, and is chilled to room temperature, and thin up shakes up to 50ml; B, filter with absorbent cotton, discard initial filtrating (owing to possibly have the dirt on the absorbent cotton in the initial filtrating, therefore discard need not), precision is measured 1ml filtrating, puts in the 10ml centrifuge tube, adds absolute ethyl alcohol 7ml; C, on vortex suspendible device, mixed 30 seconds, left the heart 15 minutes with per minute 3000; D, get deposition and add absolute ethyl alcohol 6ml, repeat the c step in step, get deposition again and add absolute ethyl alcohol 6ml, repeat the c step in step; E, the deposition water that d step is obtained go in the measuring bottle, and thin up shakes up to 100ml, as need testing solution.
The preparation precision of C, typical curve is measured reference substance solution 0,0.2,0.4,0.8,1.0ml; Put respectively in the tool plug test tube, respectively add water to 1.0ml, add 3% phenol solution 1.0ml again; Shake up; Add sulfuric acid 4.5ml rapidly, shake up, put and be chilled to room temperature; 0 pipe not measure reference substance solution is a blank; With photograph the spectrophotometric method of two appendix IVB records of Chinese Pharmacopoeia version in 2000, in the wavelength mensuration absorbance log of 490nm, the regression equation of the amount that calculates glucose and corresponding absorbance log.
D, mensuration precision are measured need testing solution 1.0ml; Add 3% phenol solution 1.0ml again; Shake up; Add sulfuric acid 4.5ml rapidly, shake up, put and be chilled to room temperature; With 0 pipe of C in the step is blank; Measure absorbance log according to spectrophotometric method in the wavelength of 490nm,, go out the polyoses content of test sample again by the regression equation calculation in C step according to the absorbance log that records; According to the medicine amount of taking by weighing of B step record, calculate the polyoses content of every gram capsule 's content again.
The assay method of the content of peptides in the quality standard detecting method of Armilariella tabescens pharmaceutical capsules is:
The bovine serum albumin(BSA) reference substance is got in the preparation of A, reference substance solution, adds water and processes the solution that every 1ml contains 0.3mg.
The content of Armilariella tabescens pharmaceutical capsules is got in the preparation of B, need testing solution, and porphyrize, the accurate title decide about 1g, and the record amount of taking by weighing; Place measuring bottle, add water 40ml, put in 80 ℃ of waters bath with thermostatic control and heated 30 minutes, jolting is constantly taken out, and is chilled to room temperature, and thin up shakes up to 50ml; Filter with absorbent cotton, discard initial filtrating, precision is measured 2ml filtrating, places measuring bottle, and thin up shakes up to 100ml.
The preparation of C, forint phenol test solution:
The preparation of forint phenol first liquid: a, get sodium potassium tartrate tetrahydrate 0.5g, add water 50ml dissolving, b, get copper sulphate 0.5g; Add water 60ml dissolving; C, get NaOH 10g and sodium carbonate 50g, add water 400ml dissolving, face with preceding a, b, three kinds of solution of c got respectively by 1: 1: 8 mixing.
The preparation of forint phenol test solution second liquid: with 8 times of above-mentioned forint phenol first liquid thin ups.
The preparation precision of D, typical curve is measured reference substance solution 0,0.2,0.4,0.8,1.0ml; Put respectively in the tool plug test tube; Respectively add water to 1.0ml, add forint phenol test solution first liquid 1.0ml again, shake up; Room temperature was placed after 10 minutes; Add forint phenol test solution second liquid 4.0ml, shake up, insulation is 5 minutes in 55 ℃ of water-baths; Take out, put and be chilled to room temperature; 0 pipe not measure reference substance solution is blank, according to spectrophotometric method in the wavelength of 650nm mensuration absorbance log, and the regression equation of the amount that calculates bovine serum albumin(BSA) and corresponding absorbance log.
E, mensuration precision are measured need testing solution 1.0ml, add forint phenol test solution first liquid 1.0ml again, shake up, and room temperature was placed after 10 minutes, add forint phenol test solution second liquid 4.0ml, shake up, and insulation is 5 minutes in 55 ℃ of water-baths, takes out, and puts and is chilled to room temperature; With 0 pipe of D in the step is blank, measures absorbance log with the photograph spectrophotometric method of two appendix IVB records of Chinese Pharmacopoeia version in 2000 in the wavelength of 650nm, is gone out the content of peptides of need testing solution again by the regression equation calculation in D step; According to the medicine amount of taking by weighing of B step record, calculate the content of peptides of every gram capsule 's content again.
The discrimination method one of the Armilariella tabescens pharmaceutical capsules that this is routine is: get capsule 's content 1g, and adding distil water 50ml heating for dissolving, absorbent cotton filters; Get filtrating 2ml, add ninhydrin solution 0.5ml, after the heating, be bluish violet and judge that then content contains the Armilariella tabescens medicine.
The discrimination method two of the Armilariella tabescens pharmaceutical capsules that this is routine is:
A, get the capsule 's content porphyrize, take by weighing 1g, put in the 50ml measuring bottle, add water 40ml, put in 80 ℃ of waters bath with thermostatic control heating 30 minutes, jolting is constantly taken out, and puts and is chilled to room temperature, and thin up shakes up to 50ml.
B, the solution that A is gone on foot filter with absorbent cotton, discard initial filtrating, and precision is measured 1ml filtrating, puts in the 10ml centrifuge tube, adds absolute ethyl alcohol 7ml, on vortex suspendible device, mixes 30 seconds, leaves the heart 15 minutes with per minute 3000.
C, the deposition of getting B step adds water 2ml makes dissolving, moves in the test tube, adds 2 of 15% methyl naphthol ethanolic solutions, and test tube wall adds the 1.0ml concentrated sulphuric acid, and two liquid meet the place, boundary if become purplish red colour circle, judge that then content contains the Armilariella tabescens medicine.
The discrimination method three of the Armilariella tabescens pharmaceutical capsules that this is routine is:
A, get capsule 's content 1.5g, add water 50ml, boiled 15 minutes, filter with absorbent cotton while hot, filtrating is put cold, and the 20ml jolting that adds diethyl ether is extracted, and obtains ether solution, filters with absorbent cotton, and filtrating volatilizes, and residue adds absolute ethyl alcohol 0.5ml dissolving, as need testing solution.
B, get the armillarisin A reference substance in addition, add absolute ethyl alcohol and process the solution that every 1ml contains 10u g, as reference substance solution.
C, according to according to thin-layered chromatography test; Draw above-mentioned A, two kinds of each 10ul of solution in B step, put respectively on same silica gel g thin-layer plate, be 8: 2: 0.5 with proportioning: the upper strata liquid of normal butyl alcohol-methyl alcohol of 10-strong ammonia solution-water is developping agent; Launch; Take out, dry, under ultraviolet lamp (365nm), inspect; In the test sample chromatogram; With the corresponding position of reference substance chromatogram on, be the fluorescence spot of same color, then judge to supply to contain the Armilariella tabescens medicine in the examination capsule 's content.
Embodiment two
This routine Armilariella tabescens pharmaceutical capsules is identical with embodiment one.The quality standard detecting method of Armilariella tabescens pharmaceutical capsules and discrimination method, slightly different with embodiment one:
The quality standard method of inspection is divided into two contents of mensuration of polyoses content and content of peptides.
Wherein measurement of the polysaccharide content step is:
The learn from else's experience 102 ℃ anhydrous dextrose reference substance that is dried to constant weight of the preparation of A, reference substance solution adds water and processes the solution that every 1ml contains anhydrous dextrose 0.08mg.
The preparation a of B, need testing solution, get the content of Armilariella tabescens pharmaceutical capsules, porphyrize, accurately claim decide about 1g and write down the amount of taking by weighing; Put in the measuring bottle, add water 35ml, put in 75 ℃ of waters bath with thermostatic control and heated 25 minutes, jolting is constantly taken out, and is chilled to room temperature, and thin up shakes up to 50ml; B, filter with absorbent cotton, discard initial filtrating, precision is measured 1ml filtrating, puts in the 10ml centrifuge tube, adds absolute ethyl alcohol 6.5ml; C, on vortex suspendible device, mixed 35 seconds, left the heart 12 minutes with per minute 2500; D, get deposition and add absolute ethyl alcohol 5.5ml, repeat the c step in step, get deposition again and add absolute ethyl alcohol 5.5ml, repeat the c step in step; E, the deposition water that d step is obtained go in the measuring bottle, and thin up shakes up to 100ml, as need testing solution.
The preparation precision of C, typical curve is measured reference substance solution 0,0.2,0.4,0.8,1.0ml; Put respectively in the tool plug test tube; Respectively add water to 1.0ml, add 3% phenol solution 0.8ml again, shake up; Add sulfuric acid 4ml rapidly; Shaking up, put and be chilled to room temperature, is blank with 0 pipe of not measuring reference substance solution; Measure absorbance log according to spectrophotometric method in the wavelength of 490nm, the regression equation of the amount that calculates glucose and corresponding absorbance log.
D, mensuration precision are measured need testing solution 1.0ml; Add 3% phenol solution 0.8ml again; Shake up; Add sulfuric acid 4.5ml rapidly, shake up, put and be chilled to room temperature; With 0 pipe of C in the step is blank; Measure absorbance log according to spectrophotometric method in the wavelength of 490nm,, go out the polyoses content of test sample again by the regression equation calculation in C step according to the absorbance log that records; According to the medicine amount of taking by weighing of B step record, calculate the polyoses content of every gram capsule 's content again.
The assay method of the content of peptides in the quality standard detecting method of Armilariella tabescens pharmaceutical capsules is:
The bovine serum albumin(BSA) reference substance is got in the preparation of A, reference substance solution, adds water and processes the solution that every 1ml contains 0.25mg.
The content of Armilariella tabescens pharmaceutical capsules is got in the preparation of B, need testing solution, and porphyrize, the accurate title decide about 1g, and the record amount of taking by weighing; Place measuring bottle, add water 35ml, put in 75 ℃ of waters bath with thermostatic control and heated 25 minutes, jolting is constantly taken out, and is chilled to room temperature, and thin up shakes up to 50ml; Filter with absorbent cotton, discard initial filtrating, precision is measured 2ml filtrating, places measuring bottle, and thin up shakes up to 100ml.
The preparation of C, forint phenol test solution:
The preparation of forint phenol first liquid: a, get sodium potassium tartrate tetrahydrate 0.4g, add water 50ml dissolving, b, get copper sulphate 0.6g; Add water 60ml dissolving; C, get NaOH 12g and sodium carbonate 45g, add water 400ml dissolving, face with preceding a, b, three kinds of solution of c got respectively by 1: 1: 8 mixing.
The preparation of forint phenol test solution second liquid: with above-mentioned forint phenol first liquid 8 times of thin ups by volume.
The preparation precision of D, typical curve is measured reference substance solution 0,0.2,0.4,0.8,1.0ml; Put respectively in the tool plug test tube; Respectively add water to 1.0ml, add forint phenol test solution first liquid 1.0ml again, shake up; Room temperature was placed after 8 minutes; Add forint phenol test solution second liquid 4.0ml, shake up, insulation is 4 minutes in 50 ℃ of water-baths; Take out, put and be chilled to room temperature; 0 pipe not measure reference substance solution is blank, according to spectrophotometric method in the wavelength of 650nm mensuration absorbance log, and the regression equation of the amount that calculates bovine serum albumin(BSA) and corresponding absorbance log.
E, mensuration precision are measured need testing solution 1.0ml, add forint phenol test solution first liquid 1.0ml again, shake up, and room temperature was placed after 8 minutes, add forint phenol test solution second liquid 4.0ml, shake up, and insulation is 4 minutes in 50 ℃ of water-baths, takes out, and puts and is chilled to room temperature; With 0 pipe of D in the step is blank, measures absorbance log according to spectrophotometric method in the wavelength of 650nm, is gone out the content of peptides of need testing solution again by the regression equation calculation in D step; According to the medicine amount of taking by weighing of B step record, calculate the content of peptides of every gram capsule 's content again.
The discrimination method one of the Armilariella tabescens pharmaceutical capsules that this is routine is: get capsule 's content 1g, and adding distil water 40ml heating for dissolving, absorbent cotton filters; Get filtrating 2.3ml, add ninhydrin solution 0.5ml, after the heating, be bluish violet and judge that then content contains the Armilariella tabescens medicine.
The discrimination method two of the Armilariella tabescens pharmaceutical capsules that this is routine is:
A, get the capsule 's content porphyrize, take by weighing 0.9g, put in the 50ml measuring bottle, add water 35ml, put in 75 ℃ of waters bath with thermostatic control heating 25 minutes, jolting is constantly taken out, and puts and is chilled to room temperature, and thin up shakes up to 50ml.
B, the solution that A is gone on foot filter with absorbent cotton, discard initial filtrating, and precision is measured 1ml filtrating, puts in the 10ml centrifuge tube, adds absolute ethyl alcohol 6.5ml, on vortex suspendible device, mixes 25 seconds, leaves the heart 12 minutes with per minute 2500.
C, the deposition of getting B step adds water 2ml makes dissolving, moves in the test tube, adds 1 of 15% methyl naphthol ethanolic solution, and test tube wall adds the 1.5ml concentrated sulphuric acid, and two liquid meet the place, boundary if become purplish red colour circle, judge that then content contains the Armilariella tabescens medicine.
The discrimination method three of the Armilariella tabescens pharmaceutical capsules that this is routine is: A, get capsule 's content 1.5g; Add water 40ml; Boiled 12 minutes, and filtered with absorbent cotton while hot, filtrating is put cold; The 18ml jolting that adds diethyl ether is extracted; Obtain ether solution, filter with absorbent cotton, filtrating volatilizes; Residue adds absolute ethyl alcohol 0.5ml dissolving, as need testing solution.
B, get the armillarisin A reference substance in addition, add absolute ethyl alcohol and process the solution that every 1ml contains 8u g, as reference substance solution.
C, according to according to thin-layered chromatography test; Draw above-mentioned A, each 12u l of two kinds of solution in B step, put respectively on same silica gel g thin-layer plate, be 8: 2: 0.5 with proportioning: the upper strata liquid of normal butyl alcohol-methyl alcohol of 10-strong ammonia solution-water is developping agent; Launch; Take out, dry, under ultraviolet lamp (365nm), inspect; In the test sample chromatogram; With the corresponding position of reference substance chromatogram on, be the fluorescence spot of same color, then judge to supply to contain the Armilariella tabescens medicine in the examination capsule 's content.
Embodiment three
This routine Armilariella tabescens pharmaceutical capsules is identical with embodiment one.The quality standard detecting method of Armilariella tabescens pharmaceutical capsules and discrimination method, slightly different with embodiment one:
The quality standard method of inspection is divided into two contents of mensuration of polyoses content and content of peptides.
Wherein measurement of the polysaccharide content step is:
The learn from else's experience 110 ℃ anhydrous dextrose reference substance that is dried to constant weight of the preparation of A, reference substance solution adds water and processes the solution that every 1ml contains anhydrous dextrose 0.12mg.
The preparation a of B, need testing solution, get the content of Armilariella tabescens pharmaceutical capsules, porphyrize, accurately claim decide about 1g and write down the amount of taking by weighing; Put in the measuring bottle, add water 45ml, put in 85 ℃ of waters bath with thermostatic control and heated 35 minutes, jolting is constantly taken out, and is chilled to room temperature, and thin up shakes up to 50ml; B, filter with absorbent cotton, discard initial filtrating, precision is measured 1ml filtrating, puts in the 10ml centrifuge tube, adds absolute ethyl alcohol 7.5ml; C, on vortex suspendible device, mixed 25 seconds, left the heart 18 minutes with per minute 3000; D, get deposition and add absolute ethyl alcohol 6.5ml, repeat the c step in step, get deposition again and add absolute ethyl alcohol 6.5ml, repeat the c step in step; E, the deposition water that d step is obtained go in the measuring bottle, and thin up shakes up to 100ml, as need testing solution.
The preparation precision of C, typical curve is measured reference substance solution 0,0.2,0.4,0.8,1.0ml; Put respectively in the tool plug test tube; Respectively add water to 1.0ml, add 3% phenol solution 1.3ml again, shake up; Add sulfuric acid 5ml rapidly; Shaking up, put and be chilled to room temperature, is blank with 0 pipe of not measuring reference substance solution; Measure absorbance log according to spectrophotometric method in the wavelength of 490nm, the regression equation of the amount that calculates glucose and corresponding absorbance log.
D, mensuration precision are measured need testing solution 1.0ml; Add 3% phenol solution 1.2ml again; Shake up; Add sulfuric acid 4.5ml rapidly, shake up, put and be chilled to room temperature; With 0 pipe of C in the step is blank; Measure absorbance log according to spectrophotometric method in the wavelength of 490nm,, go out the polyoses content of test sample again by the regression equation calculation in C step according to the absorbance log that records; According to the medicine amount of taking by weighing of B step record, calculate the content of peptides of every gram capsule 's content again.
The assay method of the content of peptides in the quality standard detecting method of Armilariella tabescens pharmaceutical capsules is:
The bovine serum albumin(BSA) reference substance is got in the preparation of A, reference substance solution, adds water and processes the solution that every 1ml contains 0.35mg.
The content of Armilariella tabescens pharmaceutical capsules is got in the preparation of B, need testing solution, and porphyrize, the accurate title decide about 1g, and the record amount of taking by weighing; Place measuring bottle, add water 45ml, put in 85 ℃ of waters bath with thermostatic control and heated 35 minutes, jolting is constantly taken out, and is chilled to room temperature, and thin up shakes up to 50ml; Filter with absorbent cotton, discard initial filtrating, precision is measured 2ml filtrating, places measuring bottle, and thin up shakes up to 100ml.
The preparation of C, forint phenol test solution:
The preparation of forint phenol first liquid: a, get sodium potassium tartrate tetrahydrate 0.6g, add water 50ml dissolving, b, get copper sulphate 0.4g; Add water 60ml dissolving; C, get NaOH 8g and sodium carbonate 55g, add water 400ml dissolving, face with preceding a, b, three kinds of solution of c got respectively by 1: 1: 8 mixing.
The preparation of forint phenol test solution second liquid: with 8 times of above-mentioned forint phenol first liquid thin ups.
The preparation precision of D, typical curve is measured reference substance solution 0,0.2,0.4,0.8,1.0ml; Put respectively in the tool plug test tube; Respectively add water to 1.0ml, add forint phenol test solution first liquid 1.0ml again, shake up; Room temperature was placed after 12 minutes; Add forint phenol test solution second liquid 4.0ml, shake up, insulation is 7 minutes in 60 ℃ of water-baths; Take out, put and be chilled to room temperature; 0 pipe not measure reference substance solution is blank, according to spectrophotometric method in the wavelength of 650nm mensuration absorbance log, and the regression equation of the amount that calculates bovine serum albumin(BSA) and corresponding absorbance log.
E, mensuration precision are measured need testing solution 1.0ml, add forint phenol test solution first liquid 1.0ml again, shake up, and room temperature was placed after 12 minutes, add forint phenol test solution second liquid 4.0ml, shake up, and insulation is 7 minutes in 60 ℃ of water-baths, takes out, and puts and is chilled to room temperature; With 0 pipe of D in the step is blank, measures absorbance log according to spectrophotometric method in the wavelength of 650nm, is gone out the content of peptides of need testing solution again by the regression equation calculation in D step; According to the medicine amount of taking by weighing of B step record, calculate the content of peptides of every gram capsule 's content again.
The discrimination method one of the Armilariella tabescens pharmaceutical capsules that this is routine is: get capsule 's content 1g, and adding distil water 60ml heating for dissolving, absorbent cotton filters; Get filtrating 1.8ml, add ninhydrin solution 0.5ml, after the heating, be bluish violet and judge that then content contains the Armilariella tabescens medicine.
The discrimination method two of the Armilariella tabescens pharmaceutical capsules that this is routine is:
A, get the capsule 's content porphyrize, take by weighing 1.1g, put in the measuring bottle, add water 45ml, put in 85 ℃ of waters bath with thermostatic control heating 35 minutes, jolting is constantly taken out, and puts and is chilled to room temperature, and thin up shakes up to 50ml.
B, the solution that A is gone on foot filter with absorbent cotton, discard initial filtrating, and precision is measured 1ml filtrating, puts in the 10ml centrifuge tube, adds absolute ethyl alcohol 7.5ml, on vortex suspendible device, mixes 35 seconds, leaves the heart 18 minutes with per minute 3000.
C, the deposition of getting B step adds water 2ml makes dissolving, moves in the test tube, adds 3 of 15% methyl naphthol ethanolic solutions, and test tube wall adds the 0.7ml concentrated sulphuric acid, and two liquid meet the place, boundary if become purplish red colour circle, judge that then content contains the Armilariella tabescens medicine.
The discrimination method three of the Armilariella tabescens pharmaceutical capsules that this is routine is:
A, get capsule 's content 1.5g, add water 60ml, boiled 18 minutes, filter with absorbent cotton while hot, filtrating is put cold, and the 23ml jolting that adds diethyl ether is extracted, and obtains ether solution, filters with absorbent cotton, and filtrating volatilizes, and residue adds absolute ethyl alcohol 0.5ml dissolving, as need testing solution.
B, get the armillarisin A reference substance in addition, add absolute ethyl alcohol and process the solution that every 1ml contains 12ug, as reference substance solution.
C, test according to photograph the thin-layered chromatography of two appendix VB records of Chinese Pharmacopoeia version in 2000; Draw above-mentioned A, two kinds of each 8ul of solution in B step; Put respectively on same silica gel g thin-layer plate; With proportioning is 8: 2: 0.5: the upper strata liquid of normal butyl alcohol-methyl alcohol of 10-strong ammonia solution-water is developping agent; Launch; Take out; Dry; Under ultraviolet lamp (365nm), inspect; In the test sample chromatogram; With the corresponding position of reference substance chromatogram on, be the fluorescence spot of same color, then judge to supply to contain the Armilariella tabescens medicine in the examination capsule 's content.
Claims (3)
1. the discrimination method of an Armilariella tabescens pharmaceutical capsules, said Armilariella tabescens pharmaceutical capsules is made by the Armilariella tabescens fermentate, and the polyoses content in every gram content of pharmaceutical capsules is with anhydrous dextrose (C
6H
120
6) meter>=60mg, content of peptides is in bovine serum albumin(BSA)>=60mg; The discrimination method of described Armilariella tabescens pharmaceutical capsules is: get capsule 's content 1g, and adding distil water 40-60ml heating for dissolving, absorbent cotton filters; Get filtrating 1.8-2.3ml, add ninhydrin solution 0.5ml, after the heating, be bluish violet and judge that then content contains the Armilariella tabescens medicine.
2. the discrimination method of an Armilariella tabescens pharmaceutical capsules, said Armilariella tabescens pharmaceutical capsules is made by the Armilariella tabescens fermentate, and the polyoses content in every gram content of pharmaceutical capsules is with anhydrous dextrose (C
6H
12O
6) meter>=60mg, content of peptides is in bovine serum albumin(BSA)>=60mg; The discrimination method of described Armilariella tabescens pharmaceutical capsules is:
A, get the capsule 's content porphyrize, take by weighing 0.9-1.1g, put in the measuring bottle, add water 35ml-45ml, put in 75 ℃ of-85 ℃ of waters bath with thermostatic control heating 25-35 minute, jolting is constantly taken out, and puts and is chilled to room temperature, and thin up shakes up to 50ml;
B, the solution that A is gone on foot filter with absorbent cotton, discard initial filtrating, and precision is measured 1ml filtrating, puts in the 10ml centrifuge tube, adds absolute ethyl alcohol 6.5-7.5ml, on vortex suspendible device, mix 25-35 second, leave heart 12-18 minute with per minute 2500-3000;
C, the deposition of getting B step adds water 2ml makes dissolving, moves in the test tube, adds 15% methyl naphthol ethanolic solution 1-3 and drips, and test tube wall adds the 0.7-1.5ml concentrated sulphuric acid, and two liquid meet the place, boundary if become purplish red colour circle, judge that then content contains the Armilariella tabescens medicine.
3. the discrimination method of an Armilariella tabescens pharmaceutical capsules, said Armilariella tabescens pharmaceutical capsules is made by the Armilariella tabescens fermentate, and the polyoses content in every gram content of pharmaceutical capsules is with anhydrous dextrose (C
6H
12O
6) meter>=60mg, content of peptides is in bovine serum albumin(BSA)>=60mg; The discrimination method of described Armilariella tabescens pharmaceutical capsules the steps include:
A, get capsule 's content 1.5g, add water 40-60ml, boiled 12-18 minute, filter with absorbent cotton while hot; Filtrating is put cold, and the 18-23ml jolting that adds diethyl ether is extracted, and obtains ether solution, filters with absorbent cotton; Filtrating volatilizes, and residue adds absolute ethyl alcohol 0.5ml dissolving, as need testing solution;
B, get the armillarisin A reference substance in addition, add absolute ethyl alcohol and process the solution that every 1ml contains 8-12u g, as reference substance solution;
C, according to according to thin-layered chromatography test; Draw above-mentioned A, two kinds of each 8-12ul of solution in B step, put respectively on same silica gel g thin-layer plate, be 8: 2: 0.5 with proportioning: the upper strata liquid of normal butyl alcohol-methyl alcohol of 10-strong ammonia solution-water is developping agent; Launch; Take out, dry, under ultraviolet lamp (365nm), inspect; In the test sample chromatogram; With the corresponding position of reference substance chromatogram on, be the fluorescence spot of same color, then judge to supply to contain the Armilariella tabescens medicine in the examination capsule 's content.
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1569044A (en) * | 2004-05-11 | 2005-01-26 | 四川大学 | Healthy product raw liquor of armilariella tabescens for strengthening the body and protecting the liver and method for improving the formulation of armilariella tabescens |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1569044A (en) * | 2004-05-11 | 2005-01-26 | 四川大学 | Healthy product raw liquor of armilariella tabescens for strengthening the body and protecting the liver and method for improving the formulation of armilariella tabescens |
Non-Patent Citations (3)
| Title |
|---|
| 宋晓凯: "《天然药物化学》", 31 August 2004, 化学工业出版社 * |
| 崔征: "《生药学》", 31 May 1996, 中国医药科技出版社 * |
| 袁锡炳: "亮菌甲素(假密环菌甲素) 含量测定方法的研究", 《药学学报》 * |
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