CN106198810A - A kind of quality determining method of the Chinese medicine composition with treatment tumor chemoradiotherapy bone marrow depression - Google Patents
A kind of quality determining method of the Chinese medicine composition with treatment tumor chemoradiotherapy bone marrow depression Download PDFInfo
- Publication number
- CN106198810A CN106198810A CN201610662320.2A CN201610662320A CN106198810A CN 106198810 A CN106198810 A CN 106198810A CN 201610662320 A CN201610662320 A CN 201610662320A CN 106198810 A CN106198810 A CN 106198810A
- Authority
- CN
- China
- Prior art keywords
- solution
- reference substance
- chinese medicine
- medicine composition
- need testing
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/11—Pteridophyta or Filicophyta (ferns)
- A61K36/12—Filicopsida or Pteridopsida
- A61K36/126—Drynaria
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/29—Berberidaceae (Barberry family), e.g. barberry, cohosh or mayapple
- A61K36/296—Epimedium
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/42—Cucurbitaceae (Cucumber family)
- A61K36/428—Trichosanthes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
- A61K36/481—Astragalus (milkvetch)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/63—Oleaceae (Olive family), e.g. jasmine, lilac or ash tree
- A61K36/638—Ligustrum, e.g. Chinese privet
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/896—Liliaceae (Lily family), e.g. daylily, plantain lily, Hyacinth or narcissus
- A61K36/8969—Polygonatum (Solomon's seal)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0087—Galenical forms not covered by A61K9/02 - A61K9/7023
- A61K9/0095—Drinks; Beverages; Syrups; Compositions for reconstitution thereof, e.g. powders or tablets to be dispersed in a glass of water; Veterinary drenches
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/90—Plate chromatography, e.g. thin layer or paper chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
Landscapes
- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Alternative & Traditional Medicine (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses the quality determining method of a kind of Chinese medicine composition with treatment tumor chemoradiotherapy bone marrow depression, the method includes that the thin layer of the Radix Astragali, Fructus Ligustri Lucidi, Rhizoma Drynariae and Herba Epimedii is identified and the Radix Astragali, the high performance liquid chromatography content assaying method of Fructus Ligustri Lucidi.The present invention is directed to many deficiencies of prior art detection method, optimal thin-layer identification method is filtered out by great many of experiments, active component detection method (includes phase composition of flowing, detection wavelength, chromatographic column, the analysis conditions such as column temperature), experiments verify that and show, the detection method that the present invention provides, can detect and evaluate the quality of the Chinese medicine composition with treatment tumor chemoradiotherapy bone marrow depression comprehensive, objective, accurately, significant for ensureing clinical efficacy.
Description
Technical field
The present invention relates to the quality determining method of a kind of Chinese medicine composition, be specifically related to one and there is treatment tumor chemoradiotherapy
The quality determining method of the Chinese medicine composition of bone marrow depression.
Background technology
Double yellow Shengbai oral liquids are a kind of natural Chinese medicines preparations with modern advanced technologies refining refinement, by Changshu
The upper pharmaceutical Co. Ltd of Lei Yun produces, by the Radix Astragali, processed with wine Rhizoma Polygonati, processed with wine Fructus Ligustri Lucidi, ironed Rhizoma Drynariae, Radix Trichosanthis, Herba Epimedii system
Become.There is supplementing QI and nourishing YIN, the raw marrow of the kidney invigorating, be clinically used for treatment leukocyte caused by tumor chemoradiotherapy bone marrow depression and reduce disease,
Curative effect is reliable.In prior art, the detection method about double yellow Shengbai oral liquids is not reported, the detection report of the most single flavour of a drug
Road, can not comprehensively, objectively detect and evaluate the quality of double yellow Shengbai oral liquid.
Summary of the invention
Goal of the invention: present invention aim to address the deficiencies in the prior art, it is provided that one has treatment tumor chemoradiotherapy
The quality determining method of the Chinese medicine composition (double yellow Shengbai oral liquid) of bone marrow depression, this detection method can be objective, comprehensive, accurate
True evaluation has treatment leukocyte caused by tumor chemoradiotherapy bone marrow depression and reduces Chinese medicine composition (double yellow liter spoken parts in traditional operas of disease
Take liquid) quality, to control this have treatment tumor chemoradiotherapy bone marrow depression caused by leukocyte reduce disease Chinese medicine composition
The quality of thing (double yellow Shengbai oral liquid) is significant with guarantee clinical efficacy.
Technical scheme: in order to realize above technical purpose, the technical scheme that the present invention takes is:
A kind of quality determining method of the Chinese medicine composition with treatment tumor chemoradiotherapy bone marrow depression, by weight
Meter, described Chinese medicine composition is by the Radix Astragali 500～1000 parts, processed with wine Rhizoma Polygonati 500～1000 parts, processed with wine Fructus Ligustri Lucidi 250～500
Part, ironed Rhizoma Drynariae 500～1000 parts, Radix Trichosanthis 500～1000 parts, Herba Epimedii 250～500 parts make oral liquid formulations, institute
State detection method and include that the thin layer of the Radix Astragali, Fructus Ligustri Lucidi, Rhizoma Drynariae and Herba Epimedii is identified, comprise the following steps:
The thin layer of the Radix Astragali is identified, comprises the following steps:
(1) prepare need testing solution: take Chinese medicine composition oral liquid 10～20ml, shake extraction 2 with water-saturated n-butanol
～3 times, each 40～80ml, merge n-butyl alcohol liquid, fully wash with ammonia solution 2～4 times, each 40～80ml, discard ammoniacal liquor,
N-butyl alcohol liquid is evaporated, and residue adds methanol and dissolves, as need testing solution；
(2) prepare reference substance solution: take astragaloside reference substance, add methanol and make the solution that every 1ml is containing 1～5mg, as
Reference substance solution；
(3) according to thin layer chromatography (one annex VI B of " Chinese Pharmacopoeia " version in 2010) test, above-mentioned need testing solution is drawn
5 μ ls each with reference substance solution, put respectively on same silica gel g thin-layer plate, are 13: 7: 2 chloroform-methanol-water with volume ratio
Lower floor's solution be developing solvent, launch, take out, dry, spray with 10% ethanol solution of sulfuric acid, be heated to spot development clear；Supply
In test product chromatograph, on position corresponding with reference substance chromatograph, aobvious identical sepia speckle under sample daylight, under ultra-violet lamp
Aobvious identical orange-yellow fluorescence speckle；
The thin layer of Rhizoma Drynariae is identified, comprises the following steps:
(1) preparing need testing solution: take Chinese medicine composition oral liquid 10～20ml, add diethyl ether extraction 2～3 times, and each 20
～40ml, discard ether solution；Medicinal liquid adds ethyl acetate and extracts 2～3 times, each 20～40ml, and combined ethyl acetate liquid is evaporated,
Residue adds methanol and dissolves, as need testing solution；
(2) prepare reference substance solution: take naringin reference substance, add methanol and be respectively prepared the solution that every 1ml is containing 1～5mg, make
For reference substance solution；
(3) according to thin layer chromatography (one annex VI B of " Chinese Pharmacopoeia " version in 2010) test, above-mentioned need testing solution is drawn
5 μ ls each with reference substance solution, put respectively on same silica gel g thin-layer plate, with the methanol-butanone-chlorine of volume ratio 4: 6: 6: 1: 0.5
Imitative-water-formic acid is developing solvent, launches, and takes out, dries, and spray, with aluminum chloride test solution, is put and inspected under ultra-violet lamp, test sample color
In spectrum, on position corresponding with reference substance chromatograph, aobvious identical sapphire fluorescence speckle；
The thin layer of Herba Epimedii is identified, comprises the following steps:
(1) prepare need testing solution: take Chinese medicine composition oral liquid 10～20ml, add diethyl ether extraction 2～3, each 20～
40ml, discards ether solution；Medicinal liquid adds ethyl acetate and extracts 2～3 times, each 20～40ml, and combined ethyl acetate liquid is evaporated, residual
Slag adds methanol makes dissolving, as need testing solution；
(2) prepare reference substance solution: take icariin reference substance, add methanol and be respectively prepared the solution that every 1ml is containing 1～5mg,
As reference substance solution；
(3) according to thin layer chromatography (one annex VI B of " Chinese Pharmacopoeia " version in 2010) test, need testing solution and right is drawn
According to each 5 μ l of product solution, put respectively on same silica gel g thin-layer plate, with chloroform-methanol-water that volume ratio is 13: 7: 2
Lower floor's solution is developing solvent, launches, and takes out, dries, and spray, with 10% ethanol solution of sulfuric acid, is heated to spot development clear, for examination
In product chromatograph, on position corresponding with reference substance chromatograph, the speckle of aobvious identical brown color；
The thin layer of Fructus Ligustri Lucidi is identified, comprises the following steps:
(1) need testing solution is prepared: take Chinese medicine composition oral liquid 10～20ml, by D101 type macroporous adsorbent resin
Post, with water elution, discards water liquid, then with volumetric concentration 20% ethanol elution, discards eluent, continues with volumetric concentration 30% ethanol
Eluting, collects eluent, is evaporated, and residue adds ethanol makes dissolving, as need testing solution；
(2) prepare reference substance solution: take specnuezhenside reference substance, add methanol and be respectively prepared the solution that every 1ml is containing 1～5mg,
As reference substance solution；
(3) according to thin layer chromatography (one annex VI B of " Chinese Pharmacopoeia " version in 2010) test, need testing solution and right is drawn
According to each 5 μ l of product solution, put respectively on same silica GF254 lamellae, be 13: 7: 2 chloroform-methanol-water with volume ratio
Lower floor's solution be developing solvent, launch, take out, dry, lamellae is put in iodine cylinder stifling, clear to spot development, test sample
In chromatograph, on position corresponding with reference substance chromatograph, aobvious identical yellow spotting.
Preferably, the quality inspection of the above-described Chinese medicine composition with treatment tumor chemoradiotherapy bone marrow depression
Survey method, counts by weight, and described Chinese medicine composition is by the Radix Astragali 500 parts, processed with wine Rhizoma Polygonati 500 parts, processed with wine Fructus Ligustri Lucidi 250
Part, ironed Rhizoma Drynariae 500 parts, Radix Trichosanthis 500 parts, Herba Epimedii 250 parts make oral liquid formulations, and described detection method includes Huang
The thin layer of stilbene, Fructus Ligustri Lucidi, Rhizoma Drynariae and Herba Epimedii is identified, comprises the following steps:
The thin layer of the Radix Astragali is identified, comprises the following steps:
(1) prepare need testing solution: take Chinese medicine composition oral liquid 10ml, shake extraction 2 times with water-saturated n-butanol, often
Secondary 40ml, merges n-butyl alcohol liquid, fully washs with ammonia solution 2 times, each 40ml, discard ammoniacal liquor, and n-butyl alcohol liquid is evaporated, and residue adds
Methanol dissolves, as need testing solution；
(2) prepare reference substance solution: take astragaloside reference substance, add methanol and make every 1ml solution containing 1mg, as right
According to product solution；
(3) according to thin layer chromatography (one annex VI B of " Chinese Pharmacopoeia " version in 2010) test, above-mentioned need testing solution is drawn
5 μ ls each with reference substance solution, put respectively on same silica gel g thin-layer plate, are 13: 7: 2 chloroform-methanol-water with volume ratio
Lower floor's solution be developing solvent, launch, take out, dry, spray with 10% ethanol solution of sulfuric acid, be heated to 105 DEG C to spot development
Clearly；In test sample chromatograph, on position corresponding with reference substance chromatograph, aobvious identical sepia speckle under sample daylight,
Aobvious identical orange-yellow fluorescence speckle under 365nm ultra-violet lamp；
The thin layer of Rhizoma Drynariae is identified, comprises the following steps:
(1) preparing need testing solution: take Chinese medicine composition oral liquid 10ml, add diethyl ether extraction 2 times, and each 20ml discards
Ether solution；Medicinal liquid adds ethyl acetate and extracts 2 times, each 20ml, and combined ethyl acetate liquid is evaporated, and residue adds methanol and dissolves, as
Need testing solution；
(2) prepare reference substance solution: take naringin reference substance, add methanol and be respectively prepared every 1ml solution containing 1mg, as
Reference substance solution；
(3) according to thin layer chromatography (one annex VI B of " Chinese Pharmacopoeia " version in 2010) test, above-mentioned need testing solution is drawn
5 μ ls each with reference substance solution, put respectively on same silica gel g thin-layer plate, with the methanol-butanone-chlorine of volume ratio 4: 6: 6: 1: 0.5
Imitative-water-formic acid is developing solvent, launches, and takes out, dries, and spray, with aluminum chloride test solution, is put and inspected under ultra-violet lamp, test sample color
In spectrum, on position corresponding with reference substance chromatograph, aobvious identical sapphire fluorescence speckle；
The thin layer of Herba Epimedii is identified, comprises the following steps:
(1) preparing need testing solution: take Chinese medicine composition oral liquid 10ml, add diethyl ether extraction 2, and each 20ml discards second
Ether liquid；Medicinal liquid adds ethyl acetate and extracts 2 times, each 20ml, and combined ethyl acetate liquid is evaporated, and residue adds methanol makes dissolving, as
Need testing solution；
(2) prepare reference substance solution: take icariin reference substance, add methanol and be respectively prepared every 1ml solution containing 1mg, make
For reference substance solution；
(3) according to thin layer chromatography (one annex VI B of " Chinese Pharmacopoeia " version in 2010) test, need testing solution and right is drawn
According to each 5 μ l of product solution, put respectively on same silica gel g thin-layer plate, with chloroform-methanol-water that volume ratio is 13: 7: 2
Lower floor's solution is developing solvent, launches, and takes out, dries, and spray, with 10% ethanol solution of sulfuric acid, is heated to spot development clear, for examination
In product chromatograph, on position corresponding with reference substance chromatograph, the speckle of aobvious identical brown color；
The thin layer of Fructus Ligustri Lucidi is identified, comprises the following steps:
(1) need testing solution is prepared: take Chinese medicine composition oral liquid 10ml, by D101 type macroporous adsorptive resins, with
Water elution, discards water liquid, then with volumetric concentration 20% ethanol 50mL eluting, discards eluent, continues with volumetric concentration 30% ethanol
50mL eluting, collects eluent, is evaporated, and residue adds ethanol makes dissolving, as need testing solution；
(2) prepare reference substance solution: take specnuezhenside reference substance, add methanol and be respectively prepared every 1ml solution containing 1mg, make
For reference substance solution；
(3) according to thin layer chromatography (one annex VI B of " Chinese Pharmacopoeia " version in 2010) test, need testing solution and right is drawn
According to each 5 μ l of product solution, put respectively on same silica GF254 lamellae, be 13: 7: 2 chloroform-methanol-water with volume ratio
Lower floor's solution be developing solvent, launch, take out, dry, lamellae is put in iodine cylinder stifling, clear to spot development, test sample
In chromatograph, on position corresponding with reference substance chromatograph, aobvious identical yellow spotting.
Preferably, the quality inspection of the above-described Chinese medicine composition with treatment tumor chemoradiotherapy bone marrow depression
Survey method, it is characterised in that Chinese medicine composition oral liquid formulations is prepared by the following method and obtains: take by weight the Radix Astragali,
Processed with wine Rhizoma Polygonati, processed with wine Fructus Ligustri Lucidi, ironed Rhizoma Drynariae, Radix Trichosanthis, Herba Epimedii, add medical material 8～12 times amount water, decocts 2～3 times, often
Secondary 1.5～2 hours, filtering, merging filtrate is also concentrated into 60 DEG C of relative densities and is about 1.12～1.18, adds ethanol and makes alcohol content
Reaching 55～65%, 0 DEG C～4 DEG C of cold preservations 24～48 hours, filtration, filtrate tune pH to 7.0～7.2, decompression recycling ethanol is the denseest
Being reduced to 60 DEG C of relative densities is 1.30～1.40, adjust pH to 4.5,0 DEG C～4 DEG C of cold preservations 24～48 hours, filter, take steviosin
It is dissolved in water with xylitol, adds in filtrate and mix, regulate pH to 7.5～8.0, stir evenly, filter, embedding, 100 DEG C of sterilizings
30min, to obtain final product.
Preferably, the quality inspection of the above-described Chinese medicine composition with treatment tumor chemoradiotherapy bone marrow depression
Survey method, it is characterised in that described detection method also includes using the Radix Astragali in high performance liquid chromatography detection Chinese medicine composition
Content, comprises the following steps:
(1) preparation of reference substance solution: take astragaloside reference substance appropriate, accurately weighed, add methanol and make every 1ml and contain
The solution of 0.5～1mg, to obtain final product；
(2) preparation of need testing solution: precision measures Chinese medicine composition oral liquid 10ml, shakes with water saturated n-butyl alcohol
Extract 2～4 times, merge n-butyl alcohol liquid, fully wash with ammonia solution 2～3 times, discard ammoniacal liquor, n-butanol extracting liquid recycling design
To dry, residue adds methanol and dissolves and be transferred in volumetric flask, adds methanol to scale, shakes up, and supernatant was centrifuged film, to obtain final product.
Chromatographic condition and system suitability: with octadecylsilane chemically bonded silica as filler；With A phase acetonitrile-B phase
Water carries out gradient elution for flowing phase according to the form below；Employing evaporative light scattering detector detects, and theoretical cam curve presses astragaloside peak
Calculating should be not less than 3000；
Algoscopy: precision draws reference substance solution 5 μ l, 15 μ l and need testing solution 10 μ l respectively, injects chromatograph of liquid,
Measure, calculate with external standard two-point method logarithmic equation, to obtain final product；
Every 1ml Chinese medicine composition oral liquid in terms of astragaloside, must not be less than 0.15mg containing the Radix Astragali.
Preferably, the quality inspection of the above-described Chinese medicine composition with treatment tumor chemoradiotherapy bone marrow depression
Survey method, it is characterised in that described detection method also includes using Fructus Ligustri Lucidi in high performance liquid chromatography detection Chinese medicine composition
Content, comprise the following steps:
(1) preparation of reference substance solution: take specnuezhenside reference substance appropriate, accurately weighed, add methanol and make every 1ml and contain
The solution of 0.1mg, to obtain final product.；
(2) preparation of need testing solution: precision measures in Chinese medicine composition oral liquid 5ml to 50ml measuring bottle, adds water to carve
Degree, shakes up, and supernatant was centrifuged film, to obtain final product；
Chromatographic condition and system suitability: with octadecylsilane chemically bonded silica as filler；With volume ratio for 19:
81 acetonitrile-waters are flowing phase；Detection wavelength is 224nm；Theoretical cam curve is calculated by specnuezhenside peak should be not less than 3000；
Algoscopy: precision draws reference substance solution and need testing solution 10 μ l respectively, injects chromatograph of liquid, measures, i.e.
?；
Every 1ml Chinese medicine composition oral liquid in terms of specnuezhenside, must not be less than 0.50mg containing Fructus Ligustri Lucidi.
Chinese medicine composition (double Huangs of disease are reduced owing to the present invention treats the leukocyte caused by tumor chemoradiotherapy bone marrow depression
Shengbai oral liquid) constitute containing 6 taste medical materials, containing highly polar in extract, middle polarity isoreactivity composition, the color of prior art
Spectrum separation condition can't reflect its inherent quality comprehensively, and the present invention can be objectively responded and evaluate in this by great many of experiments screening
The detection method of drug composition, preferably obtains optimal chromatographic condition, and composition and optimal gradient including optimal flow phase are washed
De-program, detects wavelength, flow velocity, chromatographic column and column temperature etc..Use the chromatographic condition that the present invention preferably obtains, can be to Chinese drug-treated group
In compound, each composition carries out science detection, and repeatability and stability excellent.
Beneficial effect: the Chinese medicine that the present invention provides has treatment leukocyte caused by tumor chemoradiotherapy bone marrow depression to be reduced
The quality determining method of the Chinese medicine composition of disease has the advantage that
The present invention is directed to many deficiencies of prior art detection method, filter out optimal thin layer by great many of experiments and differentiate
Method, active component detection method (includes the analysis conditions such as phase composition of flowing, detection wavelength, chromatographic column, column temperature), through testing
Card shows, the detection method that the present invention provides, and can detect comprehensive, objective, accurately and evaluation has treatment tumor chemoradiotherapy
Leukocyte caused by bone marrow depression reduces the quality of the Chinese medicine composition of disease, significant for ensureing clinical efficacy.
Detailed description of the invention
Be further elucidated with the present invention below in conjunction with specific embodiment, it should be understood that these embodiments be merely to illustrate the present invention and
It is not used in restriction the scope of the present invention, after having read the present invention, those skilled in the art's shape various of equal value to the present invention
The amendment of formula all falls within the application claims limited range.
Embodiment 1
The Chinese medicine composition oral liquid formulations (double yellow Shengbai oral liquid) 1, with treatment tumor chemoradiotherapy bone marrow depression is logical
Cross following methods to prepare: take Radix Astragali 500g, processed with wine Rhizoma Polygonati 500g, processed with wine Fructus Ligustri Lucidi 250g, ironed osteomiosis by weight
Mend 500g, Radix Trichosanthis 500g, Herba Epimedii 250, add medical material 12 times amount water, decoct 2 times, each 1.5 hours, filter, merging filtrate
And be concentrated into 60 DEG C of relative densities and be about 1.12～1.18, add ethanol and make alcohol content reach 55%, 0 DEG C～4 DEG C of cold preservations 248 little
Time, filtering, filtrate adjusts pH to 7.0, decompression recycling ethanol to be concentrated into 60 DEG C of relative densities be 1.30～1.40, adjusts pH to 4.5,
0 DEG C～4 DEG C of cold preservation 48 hours, filter, take steviosin and xylitol is dissolved in water, add in filtrate and mix, regulation pH to 7.5～
8.0, stir evenly, filter, embedding, 100 DEG C of sterilizing 30min, to obtain final product.
2, instrument and reagent
MS105DU, AL204, MT5 electronic balance (Mei Tele company)；ZF-20C dark box type uv analyzer (Shanghai Baoshan
Turn round and look at electric light instrument plant of village)；KQ-100B type ultrasonic cleaner (Kunshan ultrasonic instrument company limited)；DHG-9070A type electric heating is permanent
Temperature air dry oven (above Nereid's grand experimental facilities company limited).
Astragaloside is provided (lot number: 110781-200613) by Nat'l Pharmaceutical & Biological Products Control Institute, for assay
With；Specnuezhenside is provided (lot number: 111926-201203) by Nat'l Pharmaceutical & Biological Products Control Institute, for assay；Excessive sheep
Icariin is provided (lot number: 110737-200415) by Nat'l Pharmaceutical & Biological Products Control Institute, for assay；Naringin by
State's medicine biological products assay institute provides (lot number: 110722-201111), for assay；D101 type macroporous resin (Nanjing
Nan Ao company limited)；Reagent (the Nanjing chemical reagent such as n-butyl alcohol, ammonia, ether, ethyl acetate, chloroform, ethanol, methanol
Company limited)；Lamellae (Qingdao Marine Chemical Co., Ltd.).
3, the quality determining method of a kind of Chinese medicine composition with treatment tumor chemoradiotherapy bone marrow depression, detection method bag
The thin layer including the Radix Astragali, Fructus Ligustri Lucidi, Rhizoma Drynariae and Herba Epimedii is identified, comprises the following steps:
(1) discriminating of the Radix Astragali
The preparation of need testing solution: take above Chinese medicine composition oral liquid formulations 10ml, carries with water-saturated n-butanol shaking
Taking 3 times, each 40ml, merge n-butyl alcohol liquid, fully wash with ammonia solution 2 times, each 40ml, discard ammoniacal liquor, n-butyl alcohol liquid steams
Dry, residue adds 5ml methanol and dissolves, as need testing solution.
The preparation of reference substance solution: separately take astragaloside reference substance, adds methanol and makes every 1ml solution containing 1mg, as right
According to product solution.
The preparation of negative control solution: by need testing solution preparation method, prepare without Radix Astragali negative control solution with method.
Thin layer chromatography: according to thin layer chromatography (" Chinese Pharmacopoeia " 2010 editions one annex VI B) test, draw above-mentioned three kinds molten
The each 5 μ l of liquid, put respectively on same silica gel g thin-layer plate, with lower floor's solution of chloroform-methanol-water (13: 7: 2) for launching
Agent, launches, and takes out, dries, and spray, with 10% ethanol solution of sulfuric acid, is heated to spot development at 105 DEG C clear.
Result: in test sample chromatograph, on position corresponding with reference substance chromatograph, aobvious identical sepia under sample daylight
Speckle, aobvious identical orange-yellow fluorescence speckle under ultra-violet lamp (365nm), and negative immaculate.The results are shown in Table 1.
1 pair, table yellow Shengbai oral liquid Radix Astragali discrimination test result
Conclusion: verify through ten batch samples, this method differentiates that in preparation, Radix Astragali specificity is good, and favorable reproducibility, negative control is without dry
Disturb, differentiate to list quality standard text in by the thin layer of the Radix Astragali.
(2) discriminating of Rhizoma Drynariae
The preparation of need testing solution: take above Chinese medicine composition oral liquid formulations 10ml, add diethyl ether extraction 2 times, every time
20ml, discards ether solution.Medicinal liquid adds ethyl acetate and extracts 2 times, each 20ml, and combined ethyl acetate liquid is evaporated, and residue adds methanol
5ml makes dissolving, as need testing solution.
The preparation of reference substance solution: separately take naringin reference substance, adds methanol and is respectively prepared every 1ml solution containing 1mg, as
Reference substance solution.
The preparation of negative control solution: by need testing solution preparation method, prepare without Rhizoma Drynariae negative control molten with method
Liquid.
Thin layer chromatography: according to thin layer chromatography (" Chinese Pharmacopoeia " 2010 editions one annex VI B) test, draw above-mentioned three kinds molten
The each 5 μ l of liquid, put respectively on same silica gel g thin-layer plate, with methanol-butanone-chloroform-water-formic acid (4: 6: 6: 1: 0.5) for launching
Agent, launches, and takes out, dries, and spray, with aluminum chloride test solution, is put and inspected under ultra-violet lamp (365nm).
Result: in test sample chromatograph, on position corresponding with reference substance chromatograph, aobvious identical sapphire fluorescence speckle,
And negative sample is without this speckle.The results are shown in Table 2.
2 pairs, table yellow Shengbai oral liquid Rhizoma Drynariae discrimination test result
Conclusion: verify through ten batch samples, this method differentiates that Rhizoma Drynariae specificity is good in preparation, favorable reproducibility, negative control without
Interference, therefore differentiate to list quality standard text in by the thin layer of Rhizoma Drynariae.
(3) discriminating of Herba Epimedii
The preparation of need testing solution: take above Chinese medicine composition oral liquid formulations 10ml, add diethyl ether extraction 2 times, every time
20ml, discards ether solution.Medicinal liquid adds ethyl acetate and extracts 2 times, each 20ml, and combined ethyl acetate liquid is evaporated, and residue adds methanol
5ml makes dissolving, as need testing solution.
The preparation of reference substance solution: separately take icariin reference substance, adds methanol and is respectively prepared every 1ml solution containing 1mg, make
For reference substance solution.
The preparation of negative control solution: by need testing solution preparation method, prepare without Herba Epimedii negative control molten with method
Liquid.
Thin layer chromatography: according to thin layer chromatography (" Chinese Pharmacopoeia " 2010 editions one annex VI B) test, draw above-mentioned three kinds molten
The each 5 μ l of liquid, put respectively on same silica gel g thin-layer plate, with lower floor's solution of chloroform-methanol-water (13: 7: 2) for launching
Agent, launches, and takes out, dries, and spray, with 10% ethanol solution of sulfuric acid, is heated to spot development at 110 DEG C clear.
Result: in test sample chromatograph, on position corresponding with reference substance chromatograph, the speckle of aobvious identical brown color, and cloudy
Property is without this speckle.The results are shown in Table 3.
3 pairs, table yellow Shengbai oral liquid Herba Epimedii discrimination test result
Conclusion: verify through ten batch samples, this method differentiates that Herba Epimedii specificity is good in preparation, favorable reproducibility, negative control without
Interference, therefore differentiate to list quality standard text in by the thin layer of Herba Epimedii.
(4) discriminating of Fructus Ligustri Lucidi
The preparation of need testing solution: take above Chinese medicine composition oral liquid formulations 10ml, by D101 type macroporous absorption tree
Fat post (internal diameter 1.5cm, a height of 12cm of post), with water 100ml eluting, discards water liquid, then with 20% ethanol 50ml eluting, discards and wash
De-liquid, continues with 30% ethanol 50ml eluting, collects eluent, be evaporated, and residue adds 30% ethanol 5ml makes dissolving, as test sample
Solution.
The preparation of reference substance solution: separately take specnuezhenside reference substance, adds methanol and is respectively prepared every 1ml solution containing 1mg, make
For reference substance solution.
The preparation of negative control solution: by need testing solution preparation method, prepare without Fructus Ligustri Lucidi negative control molten with method
Liquid.
Thin layer chromatography: according to thin layer chromatography (" Chinese Pharmacopoeia " 2010 editions one annex VI B) test, draw above-mentioned three kinds molten
The each 5 μ l of liquid, put respectively on same silica GF254 lamellae, with lower floor's solution of chloroform-methanol-water (13: 7: 2) are
Developing solvent, launches, and takes out, dries, put and inspect under ultra-violet lamp (254nm), and is put by lamellae in iodine cylinder stifling.
Result: in test sample chromatograph, on position corresponding with reference substance chromatograph, aobvious identical brown speckle, and negative nothing
This speckle.Lamellae is put in iodine cylinder stifling, clear to spot development, in test sample chromatograph, corresponding with reference substance chromatograph
On position, aobvious identical yellow spotting, and feminine gender is without this speckle；Integrated comparative finds to use the colour developing of iodine cylinder relatively sharp substantially, therefore
The colour developing of iodine cylinder is listed in quality standard as colour developing mode.
The results are shown in Table 4.
4 pairs, table yellow Shengbai oral liquid Fructus Ligustri Lucidi discrimination test result
Conclusion: verify through ten batch samples, this method differentiates that in preparation, the specificity of Fructus Ligustri Lucidi is good, favorable reproducibility, negative control
Noiseless, therefore differentiate to list quality standard text in by the thin layer of Fructus Ligustri Lucidi.
4, the assay of astragaloside in Chinese medicine composition
(1) preparation of reference substance solution
Take astragaloside reference substance appropriate, accurately weighed, add methanol and make every 1ml solution containing 0.512mg, to obtain final product.
(2) preparation of need testing solution
Precision measures above Chinese medicine composition oral liquid formulations 10ml, shakes extraction 4 times with water saturated n-butyl alcohol, every time
40ml, merges n-butyl alcohol liquid, fully washs with ammonia solution 2 times, each 40ml, discard ammoniacal liquor, n-butanol extracting liquid recycling design
To dry, residue adds methanol and dissolves and be transferred in 10ml volumetric flask, adds methanol to scale, shakes up, and supernatant crosses film, to obtain final product.
(3) chromatographic condition
Chromatographic column: Waters Symmetry-C18(4.6×250mm,5μm)；Press mutually with acetonitrile (A)-water (B) for flowing
Table carries out gradient elution, such as table 5；Employing evaporative light scattering detector detects.
Table 5 gradient elution table
(4) specificity is investigated
Astragaloside and the separating degree of other components are more than 1.5 with this understanding, and peak shape is good, and theoretical cam curve is with the Radix Astragali
First glycosides peak meter is not less than 3000, and negative sample is without astragaloside peak.Result shows that this method system suitability is good.
(5) linear relationship is investigated
Precision weighs astragaloside reference substance, adds methanol and is made into every milliliter of reference substance solution containing astragaloside 512 μ g；
Accurate absorption 2,5,10,15,20 μ l reference substance solution, are measured by above-mentioned chromatographic condition sample introduction, record peak area.With sample size (μ
G) common logarithm is abscissa, and the common logarithm of peak area is vertical coordinate, calculates regression equation and is: Y=1.9822X+
4.8698, r=0.9989.Result shows: astragaloside between 1.02～10.24 μ g in good linear relationship.(it is shown in Table
6)。
Table 6 linear relationship is investigated
(6) precision is investigated
The same reference substance solution of accurate absorption, continuous sample introduction 6 times, record peak area, RSD is 1.42%, and result shows essence
Density is good.Such as table 7
Table 7 precision test
(7) study on the stability
Taking Radix Astragali sample solution, room temperature is placed, and carries out METHOD FOR CONTINUOUS DETERMINATION every 4h, measures 6 times, during with the reservation of astragaloside
Between and mass concentration evaluate sample stability, calculate RSD, result shows that sample stability is good.Such as table 8
Table 8 stability test
(8) repeatability is investigated
The same a collection of Chinese medicine composition oral liquid (double yellow Shengbai oral liquid) that the method as described in above 1 that takes prepares,
Preparing sample solution 6 parts with method, under above-mentioned chromatographic condition, sample introduction measures respectively, the average quality of astragaloside in results sample
Concentration be 0.41mg/ml, RSD be 4.24%, show that repeatability is good.Such as table 9.
Table 9 replica test
(9) accuracy is investigated
The accurate concentration known sample solution 2.5ml that draws in volumetric flask, accurate adds astragaloside reference substance solution 2ml
(concentration 0.635mg/ml), is diluted with water to 10ml, prepares 6 parts of samples by above-mentioned test sample preparation method, and sample residue adds first
Alcohol dissolves, and is settled in 5ml volumetric flask, sampling and measuring, calculates the response rate and RSD；Result mean sample recovery rate is
100.53%, RSD are 2.33%, show that this method accuracy is good.Such as table 10
Table 10 accuracy is investigated
The average recovery rate of result astragaloside is 100.53%, RSD=2.33% (n=6).
(10) sample size measures
The Chinese medicine composition oral liquid formulations (double yellow Shengbai oral liquid) 7 batches that method as described in above 1 prepares is little
Try and test agent in 3 batches, every batch parallel 2 parts, prepare sample by test sample sample preparation methods, sample introduction under above-mentioned chromatographic condition
Measuring, accurate pipette samples solution, sample introduction measures, calculates the concentration of astragaloside in every batch sample.Result such as table 11.
Table 11 assay result (n=2)
(11) conclusion
The rate of transform of astragaloside in double yellow Shengbai oral liquid lab scale seven batch sample, pilot-scale three batch sample preparation process
Basicly stable, between 63.53%～78.85%, because of in the different batches Radix Astragali decoction pieces fluctuation of Astragaloside content and cause
The content of astragaloside slightly difference in sample.According to above Chinese medicine composition oral liquid formulations 10 batch sample Astragaloside content
Measurement result (0.21～0.41mg/ml), floats downward 30% by astragaloside by content minimum, specifies above Chinese medicine composition temporarily
Oral liquid formulations every milliliter oral liquid contains the Radix Astragali with astragaloside (C41H68O14) meter, 0.15mg must not be less than.
5, the assay of specnuezhenside in Chinese medicine composition
(1) preparation of reference substance solution
Take specnuezhenside reference substance appropriate, accurately weighed, add methanol and make every 1ml and contain the right of specnuezhenside 382.7 μ g
According to product stock solution, to obtain final product.
(2) preparation of need testing solution
Dilute 5 times of samples to prepare: precision measures in above Chinese medicine composition oral liquid formulations 10ml to 50ml measuring bottle, adds water
To scale, shaking up, supernatant was centrifuged film, to obtain final product；
Dilute 10 times of samples to prepare: precision measures in above Chinese medicine composition oral liquid formulations 5ml to 50ml measuring bottle, adds water
To scale, shaking up, supernatant was centrifuged film, to obtain final product；
(3) chromatographic condition
Chromatographic column: Waters sunfire-C18(4.6×250mm 5μm)；Acetonitrile-water (19:81) is flowing phase；Flow velocity:
1.0ml/min；Column temperature: 25 DEG C；Detection wavelength: 224nm.
(4) specificity is investigated
Number of theoretical plate should be not less than 3000 in terms of specnuezhenside peak, tailing factor 1.05, under the conditions of this specnuezhenside and its
The separating degree of its component should be not less than 1.5；Negative sample is without specnuezhenside peak.Result shows that this method system suitability is good.
(5) linear relationship is investigated
The accurate reference substance stock solution of different volumes of drawing, to volumetric flask, adds methanol constant volume, is prepared as every milliliter respectively
Containing the reference substance solution of 38.27,76.54,153.08,191.35,382.7 μ g, measure by above-mentioned chromatographic condition sample introduction 10 μ l
Peak area.With integrating peak areas value as abscissa, reference substance quality is vertical coordinate, calculates regression equation and is: Y=11133X-
64127, r=0.9999.Result shows, specnuezhenside between 38.28～382.7 μ g/ml in good linear relationship.Such as table
12.
Table 12 linear relationship is investigated
(6) precision is investigated
The same reference substance solution of accurate absorption, respectively sample introduction 6 times, record peak area, and RSD is 1.09%.Such as table 13
Table 13 precision test
(7) repeatability is investigated
The same a collection of Chinese medicine composition oral liquid (double yellow Shengbai oral liquid) that the method as described in above 1 that takes prepares,
With method preparation 5 times of sample solutions of dilution 6 parts, under above-mentioned chromatographic condition, sample introduction measures respectively, specnuezhenside in results sample
Average quality concentration is 228.47 μ g, and RSD is 1.93%, shows that repeatability is good.Such as table 14.
Table 14 replica test
(8) study on the stability
Accurate 5 times of sample solutions of dilution of drawing, room temperature placement, measure every 4h continuous sample introduction, measure 6 times, record face, peak
Long-pending, the RSD of result specnuezhenside peak area is 2.04%.Such as table 15.
Table 15 stability test
(9) accuracy is investigated
In double yellow Shengbai oral liquid 5ml to the 50ml volumetric flask of accurate absorption, add water constant volume, prepares 10 times of samples of dilution, takes
Dilute in 10 times of sample 1.5ml to 5ml volumetric flasks, accurate addition specnuezhenside reference substance solution 2ml (concentration 76.54 μ g/ml),
Adding 50% methanol constant volume, 6 parts of samples of parallel preparation, sample introduction measures, and calculates the response rate and RSD.The average of result specnuezhenside returns
Yield is 104.56%, RSD=1.50% (n=6), shows that this method accuracy is good.Such as table 16.
Table 16 average recovery table
(10) sample size measures
Under above-mentioned chromatographic condition, accurate pipette samples solution, sample introduction measures, and calculates the dense of specnuezhenside in every batch sample
Degree.Such as table 17.
Table 17 assay result
(11) conclusion
Chinese medicine composition oral liquid (double yellow Shengbai oral liquid) lab scale seven batch sample, pilot-scale three batch sample preparation process
The rate of transform of middle specnuezhenside is basicly stable, between 33.04%～44.44%, because of female special in different batches Fructus Ligustri Lucidi decoction pieces
The fluctuation of loyal glycosides content and cause the content of specnuezhenside slightly difference in sample.According to above Chinese medicine composition oral liquid formulations
10 batch samples specnuezhenside assay result (0.71～1.20mg/ml), float downward 30% by specnuezhenside by content minimum,
Specify that above every milliliter of oral liquid of Chinese medicine composition oral liquid formulations contains Fructus Ligustri Lucidi with specnuezhenside (molecular formula: C31H42O17) meter,
0.50mg must not be less than.
The above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For Yuan, under the premise without departing from the principles of the invention, it is also possible to make some improvements and modifications, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (7)
1. there is a quality determining method for the Chinese medicine composition for the treatment of tumor chemoradiotherapy bone marrow depression, count by weight,
Described Chinese medicine composition by the Radix Astragali 500～1000 parts, processed with wine Rhizoma Polygonati 500～1000 parts, processed with wine Fructus Ligustri Lucidi 250～500 parts, scald
Rhizoma Drynariae (processed) 500～1000 parts, Radix Trichosanthis 500～1000 parts, Herba Epimedii 250～500 parts make oral liquid formulations, and its feature exists
In, described detection method includes that the thin layer of the Radix Astragali, Fructus Ligustri Lucidi, Rhizoma Drynariae and Herba Epimedii is identified, comprises the following steps:
The thin layer of the Radix Astragali is identified, comprises the following steps:
(1) prepare need testing solution: take Chinese medicine composition oral liquid 10～20ml, extract 2～3 with water-saturated n-butanol shaking
Secondary, each 40～80ml, merge n-butyl alcohol liquid, fully wash with ammonia solution 2～4 times, each 40～80ml, discard ammoniacal liquor, just
Butanol liquid is evaporated, and residue adds methanol and dissolves, as need testing solution；
(2) prepare reference substance solution: take astragaloside reference substance, add methanol and make the solution that every 1ml is containing 1～5mg, as comparison
Product solution；
(3) according to thin layer chromatography (one annex VI B of " Chinese Pharmacopoeia " version in 2010) test, above-mentioned need testing solution and right is drawn
According to each 5 μ l of product solution, put respectively on same silica gel g thin-layer plate, be under 13: 7: 2 chloroform-methanol-water with volume ratio
Layer solution is developing solvent, launches, and takes out, dries, and spray, with 10% ethanol solution of sulfuric acid, is heated to spot development clear；Test sample
In chromatograph, on position corresponding with reference substance chromatograph, aobvious identical sepia speckle under sample daylight, aobvious phase under ultra-violet lamp
Same orange-yellow fluorescence speckle；
The thin layer of Rhizoma Drynariae is identified, comprises the following steps:
(1) prepare need testing solution: take Chinese medicine composition oral liquid 10～20ml, add diethyl ether extraction 2～3 times, each 20～
40ml, discards ether solution；Medicinal liquid adds ethyl acetate and extracts 2～3 times, each 20～40ml, and combined ethyl acetate liquid is evaporated, residual
Slag adds methanol and dissolves, as need testing solution；
(2) prepare reference substance solution: take naringin reference substance, add methanol and be respectively prepared the solution that every 1ml is containing 1～5mg, as right
According to product solution；
(3) according to thin layer chromatography (one annex VI B of " Chinese Pharmacopoeia " version in 2010) test, above-mentioned need testing solution and right is drawn
According to each 5 μ l of product solution, put respectively on same silica gel g thin-layer plate, with the methanol-butanone-chloroform of volume ratio 4: 6: 6: 1: 0.5-
Water-formic acid is developing solvent, launches, and takes out, dries, and spray, with aluminum chloride test solution, is put and inspected under ultra-violet lamp, test sample chromatograph
In, on position corresponding with reference substance chromatograph, aobvious identical sapphire fluorescence speckle；
The thin layer of Herba Epimedii is identified, comprises the following steps:
(1) preparing need testing solution: take Chinese medicine composition oral liquid 10～20ml, add diethyl ether extraction 2～3, each 20～40ml,
Discard ether solution；Medicinal liquid adds ethyl acetate and extracts 2～3 times, each 20～40ml, and combined ethyl acetate liquid is evaporated, and residue adds first
Alcohol makes dissolving, as need testing solution；
(2) prepare reference substance solution: take icariin reference substance, add methanol and be respectively prepared the solution that every 1ml is containing 1～5mg, as
Reference substance solution；
(3) according to thin layer chromatography (one annex VI B of " Chinese Pharmacopoeia " version in 2010) test, need testing solution and reference substance are drawn
The each 5 μ l of solution, put respectively on same silica gel g thin-layer plate, with the lower floor of chloroform-methanol-water that volume ratio is 13: 7: 2
Solution is developing solvent, launches, and takes out, dries, and spray, with 10% ethanol solution of sulfuric acid, is heated to spot development clear, test sample color
In spectrum, on position corresponding with reference substance chromatograph, the speckle of aobvious identical brown color；
The thin layer of Fructus Ligustri Lucidi is identified, comprises the following steps:
(1) need testing solution is prepared: take Chinese medicine composition oral liquid 10～20ml, by D101 type macroporous adsorptive resins, with
Water elution, discards water liquid, then with volumetric concentration 20% ethanol elution, discards eluent, continues and uses volumetric concentration 30% ethanol elution,
Collecting eluent, be evaporated, residue adds ethanol makes dissolving, as need testing solution；
(2) prepare reference substance solution: take specnuezhenside reference substance, add methanol and be respectively prepared the solution that every 1ml is containing 1～5mg, as
Reference substance solution；
(3) according to thin layer chromatography (one annex VI B of " Chinese Pharmacopoeia " version in 2010) test, need testing solution and reference substance are drawn
The each 5 μ l of solution, put respectively on same silica GF254 lamellae, are under 13: 7: 2 chloroform-methanol-water with volume ratio
Layer solution is developing solvent, launches, and takes out, dries, and is put by lamellae in iodine cylinder stifling, clear to spot development, test sample chromatograph
In, on position corresponding with reference substance chromatograph, aobvious identical yellow spotting.
The quality testing side of the Chinese medicine composition with treatment tumor chemoradiotherapy bone marrow depression the most according to claim 1
Method, counts by weight, described Chinese medicine composition by the Radix Astragali 500 parts, processed with wine Rhizoma Polygonati 500 parts, processed with wine Fructus Ligustri Lucidi 250 parts, scald
Rhizoma Drynariae (processed) 500 parts, Radix Trichosanthis 500 parts, Herba Epimedii 250 parts make oral liquid formulations, it is characterised in that described detection method bag
The thin layer including the Radix Astragali, Fructus Ligustri Lucidi, Rhizoma Drynariae and Herba Epimedii is identified, comprises the following steps:
The thin layer of the Radix Astragali is identified, comprises the following steps:
(1) prepare need testing solution: take Chinese medicine composition oral liquid 10ml, shake extraction 2 times with water-saturated n-butanol, every time
40ml, merges n-butyl alcohol liquid, fully washs with ammonia solution 2 times, each 40ml, discard ammoniacal liquor, and n-butyl alcohol liquid is evaporated, and residue adds first
Alcohol dissolves, as need testing solution；
(2) prepare reference substance solution: take astragaloside reference substance, add methanol and make every 1ml solution containing 1mg, as reference substance
Solution；
(3) according to thin layer chromatography (one annex VI B of " Chinese Pharmacopoeia " version in 2010) test, above-mentioned need testing solution and right is drawn
According to each 5 μ l of product solution, put respectively on same silica gel g thin-layer plate, be under 13: 7: 2 chloroform-methanol-water with volume ratio
Layer solution be developing solvent, launch, take out, dry, spray with 10% ethanol solution of sulfuric acid, be heated to 105 DEG C clear to spot development；
In test sample chromatograph, on position corresponding with reference substance chromatograph, aobvious identical sepia speckle under sample daylight, 365nm is purple
Aobvious identical orange-yellow fluorescence speckle under outer light modulation；
The thin layer of Rhizoma Drynariae is identified, comprises the following steps:
(1) preparing need testing solution: take Chinese medicine composition oral liquid 10ml, add diethyl ether extraction 2 times, and each 20ml discards ether
Liquid；Medicinal liquid adds ethyl acetate and extracts 2 times, each 20ml, and combined ethyl acetate liquid is evaporated, and residue adds methanol and dissolves, as examination
Product solution；
(2) prepare reference substance solution: take naringin reference substance, add methanol and be respectively prepared every 1ml solution containing 1mg, as comparison
Product solution；
(3) according to thin layer chromatography (one annex VI B of " Chinese Pharmacopoeia " version in 2010) test, above-mentioned need testing solution and right is drawn
According to each 5 μ l of product solution, put respectively on same silica gel g thin-layer plate, with the methanol-butanone-chloroform of volume ratio 4: 6: 6: 1: 0.5-
Water-formic acid is developing solvent, launches, and takes out, dries, and spray, with aluminum chloride test solution, is put and inspected under ultra-violet lamp, test sample chromatograph
In, on position corresponding with reference substance chromatograph, aobvious identical sapphire fluorescence speckle；
The thin layer of Herba Epimedii is identified, comprises the following steps:
(1) preparing need testing solution: take Chinese medicine composition oral liquid 10ml, add diethyl ether extraction 2, and each 20ml discards ether solution；
Medicinal liquid adds ethyl acetate and extracts 2 times, each 20ml, and combined ethyl acetate liquid is evaporated, and residue adds methanol makes dissolving, as examination
Product solution；
(2) prepare reference substance solution: take icariin reference substance, add methanol and be respectively prepared every 1ml solution containing 1mg, as right
According to product solution；
(3) according to thin layer chromatography (one annex VI B of " Chinese Pharmacopoeia " version in 2010) test, need testing solution and reference substance are drawn
The each 5 μ l of solution, put respectively on same silica gel g thin-layer plate, with the lower floor of chloroform-methanol-water that volume ratio is 13: 7: 2
Solution is developing solvent, launches, and takes out, dries, and spray, with 10% ethanol solution of sulfuric acid, is heated to spot development clear, test sample color
In spectrum, on position corresponding with reference substance chromatograph, the speckle of aobvious identical brown color；
The thin layer of Fructus Ligustri Lucidi is identified, comprises the following steps:
(1) need testing solution is prepared: take Chinese medicine composition oral liquid 10ml, by D101 type macroporous adsorptive resins, with washing
De-, discard water liquid, then with volumetric concentration 20% ethanol 50mL eluting, discard eluent, continue with volumetric concentration 30% ethanol 50mL
Eluting, collects eluent, is evaporated, and residue adds ethanol makes dissolving, as need testing solution；
(2) prepare reference substance solution: take specnuezhenside reference substance, add methanol and be respectively prepared every 1ml solution containing 1mg, as right
According to product solution；
(3) according to thin layer chromatography (one annex VI B of " Chinese Pharmacopoeia " version in 2010) test, need testing solution and reference substance are drawn
The each 5 μ l of solution, put respectively on same silica GF254 lamellae, are under 13: 7: 2 chloroform-methanol-water with volume ratio
Layer solution is developing solvent, launches, and takes out, dries, and is put by lamellae in iodine cylinder stifling, clear to spot development, test sample chromatograph
In, on position corresponding with reference substance chromatograph, aobvious identical yellow spotting.
The quality testing of the Chinese medicine composition with treatment tumor chemoradiotherapy bone marrow depression the most according to claim 1 and 2
Method, it is characterised in that Chinese medicine composition oral liquid formulations is prepared by the following method and obtains: take the Radix Astragali, wine by weight
Rhizoma Polygonati (processed), processed with wine Fructus Ligustri Lucidi, ironed Rhizoma Drynariae, Radix Trichosanthis, Herba Epimedii, add medical material 8～12 times amount water, decocts 2～3 times, every time
1.5～2 hours, filtering, merging filtrate is also concentrated into 60 DEG C of relative densities and is about 1.12～1.18, adds ethanol and makes alcohol content reach
To 55～65%, 0 DEG C～4 DEG C of cold preservations 24～48 hours, filtering, filtrate adjusts pH to 7.0～7.2, and decompression recycling ethanol also concentrates
Be 1.30～1.40 to 60 DEG C of relative densities, adjust pH to 4.5,0 DEG C～4 DEG C of cold preservations 24～48 hours, filter, take steviosin and
Xylitol is dissolved in water, and adds in filtrate and mixes, and regulates pH to 7.5～8.0, stirs evenly, and filters, embedding, 100 DEG C of sterilizing 30min,
Obtain.
The quality testing of the Chinese medicine composition with treatment tumor chemoradiotherapy bone marrow depression the most according to claim 1 and 2
Method, it is characterised in that described detection method also includes using in high performance liquid chromatography detection Chinese medicine composition the containing of the Radix Astragali
Amount, comprises the following steps:
(1) preparation of reference substance solution: take astragaloside reference substance appropriate, accurately weighed, add methanol make every 1ml containing 0.5～
The solution of 1mg, to obtain final product；
(2) preparation of need testing solution: precision measures Chinese medicine composition oral liquid 10ml, extracts with the shaking of water saturated n-butyl alcohol
2～4 times, merging n-butyl alcohol liquid, fully wash with ammonia solution 2～3 times, discard ammoniacal liquor, n-butanol extracting liquid recycling design is the most dry,
Residue adds methanol and dissolves and be transferred in volumetric flask, adds methanol to scale, shakes up, and supernatant was centrifuged film, to obtain final product.
Chromatographic condition and system suitability: with octadecylsilane chemically bonded silica as filler；With A phase acetonitrile-B phase water it is
Flowing carries out gradient elution mutually；Employing evaporative light scattering detector detects, and theoretical cam curve presses the calculating of astragaloside peak should not be low
In 3000；
Algoscopy: precision draws reference substance solution 5 μ l, 15 μ l and need testing solution 10 μ l respectively, injects chromatograph of liquid, surveys
Fixed, calculate with external standard two-point method logarithmic equation, to obtain final product；
Every 1ml Chinese medicine composition oral liquid in terms of astragaloside, must not be less than 0.15mg containing the Radix Astragali.
The quality testing side of the Chinese medicine composition with treatment tumor chemoradiotherapy bone marrow depression the most according to claim 4
Method, it is characterised in that condition of gradient elution is:
The quality testing of the Chinese medicine composition with treatment tumor chemoradiotherapy bone marrow depression the most according to claim 1 and 2
Method, it is characterised in that described detection method also includes using Fructus Ligustri Lucidi in high performance liquid chromatography detection Chinese medicine composition
Content, comprises the following steps:
(1) preparation of reference substance solution: take specnuezhenside reference substance appropriate, accurately weighed, add methanol and make every 1ml and contain 0.1
～the solution of 0.5mg, to obtain final product；
(2) preparation of need testing solution: precision measures in Chinese medicine composition oral liquid 5ml to 50ml measuring bottle, adds water to scale, shakes
Even, supernatant was centrifuged film, to obtain final product；
Chromatographic condition and system suitability: with octadecylsilane chemically bonded silica as filler；With acetonitrile-water for flowing
Phase；Detection wavelength is 224～240nm；
Algoscopy: precision draws reference substance solution and need testing solution 10 μ l respectively, injects chromatograph of liquid, measures, to obtain final product.
The quality testing side of the Chinese medicine composition with treatment tumor chemoradiotherapy bone marrow depression the most according to claim 6
Method, it is characterised in that described detection method also includes using in high performance liquid chromatography detection Chinese medicine composition the containing of Fructus Ligustri Lucidi
Amount, comprises the following steps:
(1) preparation of reference substance solution: take specnuezhenside reference substance appropriate, accurately weighed, add methanol and make every 1ml and contain
The solution of 0.1mg, to obtain final product；
(2) preparation of need testing solution: precision measures in Chinese medicine composition oral liquid 5ml to 50ml measuring bottle, adds water to scale, shakes
Even, supernatant was centrifuged film, to obtain final product；
Chromatographic condition and system suitability: with octadecylsilane chemically bonded silica as filler；With volume ratio for 19:81 second
Nitrile-water is flowing phase；Detection wavelength is 224nm；Theoretical cam curve is calculated by specnuezhenside peak should be not less than 3000；
Algoscopy: precision draws reference substance solution and need testing solution 10 μ l respectively, injects chromatograph of liquid, measures, to obtain final product；
Every 1ml Chinese medicine composition oral liquid in terms of specnuezhenside, must not be less than 0.50mg containing Fructus Ligustri Lucidi.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610662320.2A CN106198810B (en) | 2016-08-12 | 2016-08-12 | A kind of quality determining method of the Chinese medicine composition with treatment tumor chemoradiotherapy bone marrow suppression |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610662320.2A CN106198810B (en) | 2016-08-12 | 2016-08-12 | A kind of quality determining method of the Chinese medicine composition with treatment tumor chemoradiotherapy bone marrow suppression |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106198810A true CN106198810A (en) | 2016-12-07 |
| CN106198810B CN106198810B (en) | 2018-11-20 |
Family
ID=57515137
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610662320.2A Active CN106198810B (en) | 2016-08-12 | 2016-08-12 | A kind of quality determining method of the Chinese medicine composition with treatment tumor chemoradiotherapy bone marrow suppression |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106198810B (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107796908A (en) * | 2017-10-13 | 2018-03-13 | 江苏七0七天然制药有限公司 | A kind of quality determining method of FEIQIZHONG PIAN |
| CN108362788A (en) * | 2018-01-18 | 2018-08-03 | 成都中医药大学 | A kind of steamed sealwort Quality Detection analysis method |
| CN109557192A (en) * | 2018-07-05 | 2019-04-02 | 湖北梦阳药业股份有限公司 | A kind of raw white kalgan lamb skin takes the detection method of liquid |
| CN112067738A (en) * | 2020-08-19 | 2020-12-11 | 广东一方制药有限公司 | Method for identifying dampness-resolving toxin-vanquishing composition |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101020027A (en) * | 2006-12-01 | 2007-08-22 | 广州中医药大学热带医学研究所 | Medicine composition for preventing and treating AIDS and its prepn process and use |
| CN101721612A (en) * | 2008-10-31 | 2010-06-09 | 上海润知生物科技有限公司 | Chinese medicinal composition for raising leucocytes and preparation method and application thereof |
| CN104922365A (en) * | 2015-05-27 | 2015-09-23 | 广州金域医学检验中心有限公司 | Traditional Chinese medicine composition for treating osteoporosis and detection method thereof |
-
2016
- 2016-08-12 CN CN201610662320.2A patent/CN106198810B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101020027A (en) * | 2006-12-01 | 2007-08-22 | 广州中医药大学热带医学研究所 | Medicine composition for preventing and treating AIDS and its prepn process and use |
| CN101721612A (en) * | 2008-10-31 | 2010-06-09 | 上海润知生物科技有限公司 | Chinese medicinal composition for raising leucocytes and preparation method and application thereof |
| CN104922365A (en) * | 2015-05-27 | 2015-09-23 | 广州金域医学检验中心有限公司 | Traditional Chinese medicine composition for treating osteoporosis and detection method thereof |
Non-Patent Citations (3)
| Title |
|---|
| 史秀峰等: "双黄升白颗粒絮凝沉淀与醇沉工艺的比较研究", 《上海中医药杂志》 * |
| 袁春平等: "升血调元颗粒的质量标准研究", 《中国现代中药》 * |
| 雷健等: "益生胶囊质量标准研究", 《中国药师》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107796908A (en) * | 2017-10-13 | 2018-03-13 | 江苏七0七天然制药有限公司 | A kind of quality determining method of FEIQIZHONG PIAN |
| CN108362788A (en) * | 2018-01-18 | 2018-08-03 | 成都中医药大学 | A kind of steamed sealwort Quality Detection analysis method |
| CN109557192A (en) * | 2018-07-05 | 2019-04-02 | 湖北梦阳药业股份有限公司 | A kind of raw white kalgan lamb skin takes the detection method of liquid |
| CN112067738A (en) * | 2020-08-19 | 2020-12-11 | 广东一方制药有限公司 | Method for identifying dampness-resolving toxin-vanquishing composition |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106198810B (en) | 2018-11-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101850070B (en) | Detection method for Chinese medicament Tangcao tablets | |
| CN105259295B (en) | Quality detection method for ginseng, cassia twig and poria cocos oral solution | |
| CN104502518B (en) | A kind of detection method for the treatment of the Chinese medicine preparation of baby anorexia | |
| CN106198810A (en) | A kind of quality determining method of the Chinese medicine composition with treatment tumor chemoradiotherapy bone marrow depression | |
| CN104777249B (en) | The method measuring effective ingredient amygdaloside content in cough syrup of loquat leaf | |
| CN101703611B (en) | Quality detection method of Chinese angelica oral liquid for benefiting blood | |
| CN101204434A (en) | Quality standard for thrombus dispelling pill and test method thereof | |
| CN101181339B (en) | Detection method of aidi injection preparations | |
| CN102068627A (en) | Quality control method for Chinese medicine preparation Xinnaojing tabelets | |
| CN104897811B (en) | A kind of detection method of Moschus 'Xintongning ' | |
| CN101703610A (en) | Quality detection method of Qingnao antihypertensive tablet | |
| CN101991636B (en) | Detection method of Aidi injection preparation | |
| CN106680414A (en) | Detection method of compound ardisia japonica tablet | |
| CN101317935B (en) | Rujietai formulation and quality detection method | |
| CN102000139B (en) | Detection method for Aidi injection | |
| CN101987115A (en) | Jade screen oral preparation as well as preparation method and detection method thereof | |
| CN112051352B (en) | New method for controlling quality of Fukean tablets | |
| CN101011450B (en) | Quality control method for lucid ganoderma gynostemma pentaphylla oral liquid | |
| CN114544852A (en) | Improved quality detection method for Longqing capsule | |
| CN110857939B (en) | Detection method of traditional Chinese medicine composition with sleep improvement function | |
| CN102068599B (en) | Detection method for phlegm eliminating and asthma relieving cough syrup | |
| CN110297063A (en) | A kind of quality determining method of Rhizoma Chuanxiong hair washing water extract | |
| CN104749261B (en) | A kind of method checking Fuzheng Jiedu powder quality | |
| CN109709256A (en) | Scutelloside and chlorogenic acid thin layer identify detection method in a kind of Compound Jinyinhua Granules | |
| CN110763642B (en) | Detection method of traditional Chinese medicine preparation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |