CN105784914A - Detection method for traditional Chinese medicine muskiness strong bone plasters - Google Patents
Detection method for traditional Chinese medicine muskiness strong bone plasters Download PDFInfo
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Abstract
The invention relates to a detection method for traditional Chinese medicine muskiness strong bone plasters. The detection method comprises the step of identifying rhizoma atractylodis, ephedra herb, common monkshood mother roots, kusnezoff monkshood roots, dried ginger and/or rhizoma kaempferiae. Detection is carried out with thin layer chromatography, operation is easy, convenient and fast, detection cost is low, small pollution is caused to the environment, a detection result is accurate and good in repeatability, and the method can be used for detecting traditional Chinese medicine muskiness strong bone plasters.
Description
Technical field
The present invention relates to pharmaceutical technology field, particularly to the detection method of a kind of Chinese medicine SHEXIANG ZHUANGGU GAO.
Background technology
SHEXIANG ZHUANGGU GAO is recorded in 1996 version Drug Standard of Ministry of Public Health of the Peoples Republic of China Traditional Chinese medicine historical preparation the 11st.SHEXIANG ZHUANGGU GAO has analgesia, effect of antiinflammatory.For rheumatalgia, arthralgia, lumbago, neuralgia, muscular soreness, sprain, dampen.
Bone-strengthening musk paste formulation is composed as follows: Fructus Anisi Stellati, Rhizoma Kaempferiae, Radix Aconiti, Radix Aconiti Kusnezoffii, Herba Ephedrae, Rhizoma Atractylodis, the Radix Angelicae Dahuricae, Radix Angelicae Sinensis, Rhizoma Zingiberis, artificial Moschus, Mentholum, methyl salicylate, chondroitin sulfate, Borneolum Syntheticum, diphhydramine hydrochloride, Camphora etc..SHEXIANG ZHUANGGU GAO is due to respond well, according to clinical verification, evident in efficacy, enjoys patient to favor.At present, paste containing amount is only had regulation by bone-strengthening musk extractive quality standard, wherein each composition is not carried out discrimination test and assay.The report of Camphora, Mentholum, Borneolum Syntheticum and methyl salicylate content in useful gas Chromatographic Determination SHEXIANG ZHUANGGU GAO in prior art, it does not have the report qualitatively or quantitatively determined to Chinese crude drugs such as wherein Rhizoma Atractylodis, Herba Ephedrae, Rhizoma Kaempferiae, Radix Aconiti, Radix Aconiti Kusnezoffii and Rhizoma Zingiberiss.Chinese patent literature CN201010504486 discloses the discrimination method of a kind of Herba Ephedrae, and developing solvent uses chloroform to be easily make poison reagent, and easily causes environmental pollution.The quality determining method of a kind of ageratum drop pill disclosed in Chinese patent literature CN102998411A has the discriminating of Rhizoma Atractylodis, specifically add water supersound extraction, but it is not particularly suited in the emplastrum of the present invention, Chinese patent literature CN102228553A has treated disclosed in the detection method of the pharmaceutical composition of rheumatic arthritis the discriminating of a kind of aconitine, shortcoming is to use easily system poison reagent and being applied in SHEXIANG ZHUANGGU GAO not easily to operate, easy contaminated environment and harmful.SHEXIANG ZHUANGGU GAO is the rubber plaster being most widely used, and its conventional substrate has rubber, thermoplastic elastomer, Colophonium, rosin derivative, vaseline etc., and effective ingredient not easily extracts.The quality standard of SHEXIANG ZHUANGGU GAO has much room for improvement.
Summary of the invention
For overcoming the deficiencies in the prior art, the invention provides the detection method of Rhizoma Atractylodis, Herba Ephedrae, Radix Aconiti, Radix Aconiti Kusnezoffii, Rhizoma Zingiberis and/or Rhizoma Kaempferiae in a kind of Chinese medicine SHEXIANG ZHUANGGU GAO.
Term illustrates:
SHEXIANG ZHUANGGU GAO: record in the nomenclature of drug of in 1996 version Drug Standard of Ministry of Public Health of the Peoples Republic of China Traditional Chinese medicine historical preparation the 11st.Standard number: WS3-B-2268-96。
Technical scheme is as follows:
A kind of detection method of Chinese medicine SHEXIANG ZHUANGGU GAO, including one or more in following discrimination method:
A. the discriminating of Rhizoma Atractylodis
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 2～5, shred, put in conical flask, adding volume ratio 40%～50% ultrasonic 15～30min of methanol 25～50mL, filter, filtrate is evaporated, residue adds dehydrated alcohol 2mL and dissolves, and obtains need testing solution；
(2) preparation of reference substance: take Rhizoma Atractylodis control medicinal material 0.8g, put in conical flask, adds 25～50mL volume ratio 40%～50% ultrasonic 15～30min of methanol, filters, and filtrate is evaporated, and residue adds dehydrated alcohol 2mL and dissolves, and obtains control medicinal material solution；Take atisine chloride atractydin reference substance again to add methanol and make every 1mL solution containing 0.2～0.5mg, as reference substance solution；
(3) algoscopy: draw need testing solution, control medicinal material solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with volume ratio be 8～9:1～2 60～90 DEG C of petroleum ether-acetone for developing solvent, launch, take out, drying, the ethanol solution of sulfuric acid that spray is 10% with volume ratio, it is clear to spot development to heat；In test sample chromatograph, on position corresponding with control medicinal material chromatograph and reference substance chromatograph, the speckle of aobvious same color；
B. the discriminating of Herba Ephedrae
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 2～5 and be cut into slice, adds the methanol 20～50mL of volume ratio 40%～50%, supersound process 20～30min, filters, and filtrate is evaporated to 2mL, and filtrate is as need testing solution；
(2) preparation of reference substance: take ephedrine hydrochloride reference substance, adds methanol and makes every lmL solution containing lmg, as reference substance solution；
(3) algoscopy: draw each 10～15 μ L of above two solution, put respectively on same silica gel g thin-layer plate, with the methylene chloride-methanol-ammonia of volume ratio 15～20:2～6:0.5～1 for developing solvent, launch, take out, drying, spray is with ninhydrin solution, clear to spot development 105 DEG C of heating；In test sample chromatograph, on position corresponding with reference substance chromatograph, the speckle of aobvious same color；
C. the discriminating of Radix Aconiti, Radix Aconiti Kusnezoffii
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 2～5, shred, put in conical flask, add the organic solvent 25～50mL of volume ratio 40%～50%, supersound process 20～30min, filter, filtrate is evaporated, and residue adds organic solvent 2mL and dissolves, as need testing solution；
(2) preparation of reference substance: take aconitine reference substance and add organic solvent, makes every 1mL solution containing 0.5～1mg, as reference substance solution；
(3) algoscopy: draw each 5 μ L of above two solution, put respectively on same silica gel g thin-layer plate, with volume parts n-hexane-ethyl acetate-methanol than 6～7:3～4:1 for developing solvent, put in the expansion cylinder of ammonia saturated with vapor 20～30min and launch, take out, drying, spray is with rare bismuth potassium iodide test solution；In test sample chromatograph, with on reference substance chromatograph relevant position, the speckle of aobvious same color；
D. the discriminating of Rhizoma Zingiberis
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 2～5 and be cut into slice, add volume ratio 40%～50% methanol 20～50mL, supersound process 20～40min, let cool, filter, filtrate is evaporated, residue add water 20mL dissolve, extract 2～3 times with n-butyl alcohol jolting, each 30mL, merges n-butanol extracting liquid, is evaporated, residue adds 2～dissolving of 3mL methanol, obtains need testing solution；
(2) preparation of reference substance: take Rhizoma Zingiberis control medicinal material 1g, adds volume ratio 40%～50% methanol 20～50mL, supersound process 20～40min, let cool, filter, filtrate is evaporated, residue add water 20mL dissolve, extract 2～3 times with n-butyl alcohol jolting, each 30mL, merges n-butanol extracting liquid, is evaporated, residue adds 2～dissolving of 3mL methanol, obtains control medicinal material solution；
(3) algoscopy: draw need testing solution 10 μ L, control medicinal material solution 5 μ L, put respectively on same silica gel g thin-layer plate, with volume parts than 60～90 DEG C of petroleum ether-ethyl acetate-methanol-glacial acetic acid of 18～20:1～3:0.5～1:0～1 for developing solvent, launch, take out, dry, put and inspect under 365nm ultra-violet lamp, in test sample chromatograph, with on reference substance chromatograph relevant position, the fluorescence speckle of aobvious same color；
E. the discriminating of Rhizoma Kaempferiae
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 2～5, adds volume ratio 40%～50% methanol 25～50mL, supersound process 15～30min, filters, and filtrate is evaporated, and residue adds methanol 2mL and dissolves, as need testing solution；
(2) preparation of reference substance: take Ethyl p-methoxy-silicate reference substance, adds methanol and makes every 1mL solution containing 1～5mg, as reference substance solution；
(3) algoscopy: drawing need testing solution 6 μ L, reference substance solution 3 μ L puts respectively in same silica gel G F254On lamellae, with volume parts than 8～9:1～2 n-hexane-ethyl acetate for developing solvent, launch, take out, dry, put and inspect under 254nm ultra-violet lamp, in test sample chromatograph, on position corresponding with reference substance chromatograph, the speckle of aobvious same color.
According to currently preferred, in the discriminating of described Rhizoma Atractylodis, developing solvent is the volume parts 60～90 DEG C of petroleum ether-acetone than 9:2.
According to currently preferred, in the discriminating of described Herba Ephedrae, test sample prepares the methanol that solvent for use is volume ratio 40%, and developing solvent is volume parts methylene chloride-methanol-ammonia than 20:5:0.5.
According to currently preferred, in described Radix Aconiti, the discriminating of Radix Aconiti Kusnezoffii, developing solvent is that volume parts is than 6.4:3.6:1 n-hexane-ethyl acetate-methanol.
According to currently preferred, in the discriminating of described Rhizoma Zingiberis, developing solvent is the volume parts 60～90 DEG C of petroleum ether-ethyl acetate-methanol-glacial acetic acid than 20:3:1:0.4.
According to currently preferred, in the discriminating of described Rhizoma Kaempferiae, developing solvent is the volume parts n-hexane-ethyl acetate than 9:1.
According to the present invention it is further preferred that the detection method of a kind of Chinese medicine SHEXIANG ZHUANGGU GAO, including one or more in following discrimination method:
A. the discriminating of Rhizoma Atractylodis
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 210cm2Shredding and put in 100mL conical flask, add the 30mL volume ratio 40% ultrasonic 25min of methanol, filter, filtrate is evaporated, and residue adds dehydrated alcohol 2mL and dissolves, and obtains need testing solution；
(2) preparation of reference substance: take Rhizoma Atractylodis control medicinal material 0.8g, puts in 100mL conical flask, adds the 30mL volume ratio 40% ultrasonic 25min of methanol, filters, and filtrate is evaporated, and residue adds dehydrated alcohol 2mL and dissolves, and obtains control medicinal material solution；Take atisine chloride atractydin reference substance again to add methanol and make every 1mL solution containing 0.2mg, as reference substance solution；
(3) algoscopy: draw need testing solution 20 μ L, control medicinal material solution 6 μ L, reference substance solution 2 μ L, put respectively on same silica gel g thin-layer plate, with the 60～90 of volume ratio 9:2 DEG C of petroleum ether-acetone for developing solvent, launch, take out, dry, spraying the ethanol solution of sulfuric acid with volume fraction 10%, it is clear to spot development to heat；In test sample chromatograph, on position corresponding with control medicinal material chromatograph and reference substance chromatograph, the speckle of aobvious same color；
B. the discriminating of Herba Ephedrae
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 210cm2Being cut into slice, add volume ratio 40% methanol 30mL, ultrasonic 30min, filter, filtrate is evaporated, and residue adds methanol 2mL and dissolves, as need testing solution；
(2) preparation of reference substance: separately take ephedrine hydrochloride reference substance, adds methanol and makes every lmL solution containing lmg, as reference substance solution；
(3) algoscopy: draw each 10 μ L of above two solution, put respectively on same silica gel g thin-layer plate, with the methylene chloride-methanol-ammonia of volume ratio 20:5:0.5 for developing solvent, launch, take out, dry, spray is with ninhydrin solution, clear to spot development 105 DEG C of heating；In test sample chromatograph, on position corresponding with reference substance chromatograph, aobvious identical punctation.
C. the discriminating of Radix Aconiti, Radix Aconiti Kusnezoffii
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 210cm2, shredding, put in conical flask, add volume ratio 40% methanol 40mL, supersound process 30min, filter, filtrate is evaporated, and residue adds isopropanol 2mL and dissolves, as need testing solution；
(2) preparation of reference substance: take aconitine reference substance and add isopropanol, makes every 1mL solution containing 1mg, as reference substance solution；
(3) algoscopy: draw each 5 μ L of above two solution, put respectively on same silica gel thin-layer plate, with volume parts than 6.4:3.6:1 n-hexane-ethyl acetate-methanol for developing solvent, put in the expansion cylinder of ammonia saturated with vapor 20min and launch, taking out, dry, spray is with rare bismuth potassium iodide test solution；In test sample chromatograph, with on reference substance chromatograph relevant position, the speckle of aobvious same color；
D. the discriminating of Rhizoma Zingiberis
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 210cm2It is cut into slice, adds volume ratio 40% methanol 40mL, supersound process 40min, let cool, filter, filtrate is evaporated, residue add water 20mL dissolve, with n-butyl alcohol jolting extract 2 times, each 30mL, merging n-butanol extracting liquid, be evaporated, residue adds 2mL methanol and dissolves, and obtains need testing solution；
(2) preparation of reference substance: take Rhizoma Zingiberis control medicinal material 1g, adds volume ratio 40% methanol 40mL, supersound process 40min, let cool, filter, filtrate is evaporated, residue add water 20mL dissolve, extract 2 times with n-butyl alcohol jolting, each 30mL, merges n-butanol extracting liquid, is evaporated, residue adds 2mL methanol and dissolves, and obtains control medicinal material solution；
(3) algoscopy: draw need testing solution 10 μ L, control medicinal material solution 5 μ L, put respectively on same silica gel g thin-layer plate, with the volume parts 60～90 DEG C of petroleum ether-ethyl acetate-methanol-glacial acetic acid than 20:3:1:0.4 for developing solvent, launch, take out, dry, put and inspect under 365nm ultra-violet lamp, in test sample chromatograph, with on reference substance chromatograph relevant position, the fluorescence speckle of aobvious same color；
E. the discriminating of Rhizoma Kaempferiae
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 140cm2, adding volume ratio 40% methanol 30mL, supersound process 30min, filter, filtrate is evaporated, and residue adds methanol 2mL and dissolves, as need testing solution；
(2) preparation of reference substance: take Ethyl p-methoxy-silicate reference substance, adds methanol and makes every 1mL solution containing 5mg, as reference substance solution；
(3) algoscopy: drawing need testing solution 6 μ L, reference substance solution 3 μ L puts respectively in same silica gel G F254On lamellae, with the volume parts n-hexane-ethyl acetate than 9:1 for developing solvent, launch, take out, dry, put and inspect under 254nm ultra-violet lamp, in test sample chromatograph, on position corresponding with reference substance chromatograph, the speckle of aobvious same color.
Rare iodate of the present invention must prepare by potassium test solution as follows: according to 2015 editions four preparations of Chinese Pharmacopoeia, takes bismuth subnitrate/basic bismuth nitrate 0.85g, after adding glacial acetic acid 10mL and water 40mL dissolving, to obtain final product.Take 5mL before use, add liquor kalii iodide (4 → 10) 5mL, then add glacial acetic acid 20mL, be diluted with water to 100mL, to obtain final product.
Following experimental example is used for further illustrating but is not limited to the present invention.
Experimental example 1: the identification experiment of Rhizoma Atractylodis
(1) reagent and test sample
Reference substance: Rhizoma Atractylodis control medicinal material (China pharmaceutical biological product calibrating academy), lot number: 120932-201407；Atisine chloride atractydin reference substance (China pharmaceutical biological product calibrating academy), lot number: 111924-201404.
Test sample: SHEXIANG ZHUANGGU GAO (Shandong a person of good sense Fu Ruida hygienic material company limited), lot number: 20151105,20151106,20151107, specification is 7cm × 10cm.
(2) preparation of need testing solution:
Respectively with volume ratio be 0%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 100% methanol is for Extraction solvent, find that methanol volume ratio is 40% can be proposed by effective ingredient, on gained test sample position corresponding with control medicinal material chromatograph and reference substance chromatograph, the speckle of aobvious same color.The low effective ingredient of methanol volume ratio can not carry, and organic reagent used by volume ratio height increases accordingly, runs counter to experiment original intention, therefore selecting volume ratio 40% methanol is Extraction solvent.
Take SHEXIANG ZHUANGGU GAO 210cm2Shredding and put in 100mL conical flask, add the 30mL volume ratio 40% ultrasonic 25min of methanol, filter, filtrate is evaporated, and residue adds dehydrated alcohol 2mL and dissolves, and obtains need testing solution.
(3) preparation of control medicinal material solution: take Rhizoma Atractylodis control medicinal material 0.8g, puts in 100mL conical flask, adds the 30mL volume ratio 40% ultrasonic 25min of methanol, filters, and filtrate is evaporated, and residue adds dehydrated alcohol 2mL and dissolves, and obtains control medicinal material solution；
(4) preparation of reference substance solution: take atisine chloride atractydin reference substance and add methanol and make every 1mL solution containing 0.2mg, as reference substance solution；
(5) algoscopy: draw need testing solution 15 μ L, control medicinal material solution 6 μ L, reference substance solution 2 μ L, put respectively on same silica gel g thin-layer plate, with the 60～90 of volume ratio 9:2 DEG C of petroleum ether-acetone for developing solvent, launch, take out, dry, spraying the ethanol solution of sulfuric acid with volume fraction 10%, it is clear to spot development to heat.In test sample chromatograph, on position corresponding with control medicinal material chromatograph and reference substance chromatograph, the speckle of aobvious same color.
Contrast experiment: have the discriminating of Rhizoma Atractylodis in the quality determining method of a patent documentation CN102998411A ageratum drop pill, namely adds water supersound extraction by the preparation method of test sample therein, and result the speckle of same color does not occur in the position identical with comparison.
Experimental example 2: the identification experiment of Herba Ephedrae
(1) reagent and test sample
Reference substance: ephedrine hydrochloride reference substance (China pharmaceutical biological product calibrating academy), lot number: 171241-201007.
Test sample: SHEXIANG ZHUANGGU GAO (Shandong a person of good sense Fu Ruida hygienic material company limited), lot number: 20151105,20151106,20151107, specification is 7cm × 10cm.
(2) preparation of need testing solution:
Respectively with volume ratio be 0%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 100% methanol is for Extraction solvent, find that effective ingredient can be proposed by methanol volume ratio 40%, on gained test sample position corresponding with control medicinal material chromatograph and reference substance chromatograph, the speckle of aobvious same color.The low effective ingredient of methanol volume ratio can not carry, and organic reagent used by volume ratio height increases accordingly, runs counter to experiment original intention, therefore selecting volume ratio 40% methanol is Extraction solvent.
Take SHEXIANG ZHUANGGU GAO 210cm2Being cut into slice, add volume ratio 40% methanol 30mL, ultrasonic 30min, filter, filtrate is evaporated, and residue adds methanol 2mL and dissolves, as need testing solution；
(3) preparation of reference substance: take ephedrine hydrochloride reference substance, adds methanol and makes every 1mL solution containing lmg, as reference substance solution；
(4) algoscopy: draw each 10 μ L of above two solution, put respectively on same silica gel g thin-layer plate, with the methylene chloride-methanol-ammonia of volume ratio 20:5:0.5 for developing solvent, launch, take out, dry, spray is with ninhydrin solution, clear to spot development 105 DEG C of heating.In test sample chromatograph, on position corresponding with reference substance chromatograph, aobvious identical punctation.
Developing solvent is with the methylene chloride-methanol-ammonia of volume ratio 20:5:0.5 for optimal proportion, and Rf value is 0.53, meets the requirement (0.2～0.8) to Rf value of the 2015 editions thin layer chromatographys of Chinese Pharmacopoeia.
Contrast experiment: the discrimination method Extraction solvent of Chinese patent literature CN201010504486 Herba Ephedrae is methanol, developing solvent uses chloroform, Extraction solvent of the present invention is volume ratio 40% methanol, developing solvent uses dichloromethane, decrease the use of organic reagent, do not use easily system poison reagent simultaneously, alleviate the experiment injury to laboratory personnel.
Experimental example 3: Radix Aconiti, Radix Aconiti Kusnezoffii identification experiment
(1) reagent and test sample:
Reference substance: aconitine reference substance (China pharmaceutical biological product calibrating academy), lot number: 120932-201407；
Test sample: SHEXIANG ZHUANGGU GAO (Shandong a person of good sense Fu Ruida hygienic material company limited), lot number: 20151105,20151106,20151107, specification is 7cm × 10cm.
(2) preparation of need testing solution:
With isopropanol, methanol, normal hexane as Extraction solvent, on gained test sample position corresponding with control medicinal material chromatograph and reference substance chromatograph, the speckle of aobvious same color.For further optimization experiment scheme, respectively with volume ratio be 10%, 20%, 30%, 40%, 50%, 60%, 70% methanol is for Extraction solvent, find that methanol volume ratio is 40% can be proposed by effective ingredient, on gained test sample position corresponding with control medicinal material chromatograph and reference substance chromatograph, the speckle of aobvious same color, therefore selecting volume ratio 40% methanol is Extraction solvent.
Taking SHEXIANG ZHUANGGU GAO 3, shred, put in conical flask, add volume ratio 40% methanol 40mL, supersound process 30min, filter, filtrate is evaporated, and residue adds isopropanol 2mL and dissolves, as need testing solution；
(2) preparation of reference substance: take aconitine reference substance and add isopropanol, makes every 1mL solution containing 1mg, as reference substance solution；
(3) algoscopy: draw each 5 μ L of above two solution, put respectively on same silica gel thin-layer plate, with volume parts than 6.4:3.6:1 n-hexane-ethyl acetate-methanol for developing solvent, put in the expansion cylinder of ammonia saturated with vapor 20min and launch, taking out, dry, spray is with rare bismuth potassium iodide test solution；In test sample chromatograph, with on reference substance chromatograph relevant position, the speckle of aobvious same color.
Experimental example 4: the identification experiment of Rhizoma Zingiberis
(1) reagent and test sample:
Reference substance: Rhizoma Zingiberis control medicinal material (China pharmaceutical biological product calibrating academy), lot number: 120942-200907.
Test sample: SHEXIANG ZHUANGGU GAO (Shandong a person of good sense Fu Ruida hygienic material company limited), lot number: 20151105,20151106,20151107, specification is 7cm × 10cm.
(2) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 4 and be cut into slice, add volume ratio 40%～50% methanol 40mL, supersound process 40min, let cool, filter, filtrate is evaporated, residue add water 20mL dissolve, extract 2 times with n-butyl alcohol jolting, each 30mL, merges n-butanol extracting liquid, is evaporated, residue adds 2mL methanol and dissolves, and obtains need testing solution；
(3) preparation of reference substance: take Rhizoma Zingiberis control medicinal material 1g, adds volume ratio 40%～50% methanol 40mL, supersound process 40min, let cool, filter, filtrate is evaporated, residue add water 20mL dissolve, extract 2 times with n-butyl alcohol jolting, each 30mL, merges n-butanol extracting liquid, is evaporated, residue adds 2mL methanol and dissolves, and obtains control medicinal material solution；
(4) algoscopy: draw need testing solution 10 μ L, control medicinal material solution 5 μ L, put respectively on same silica gel g thin-layer plate, with the volume parts 60～90 DEG C of petroleum ether-ethyl acetate-methanol-glacial acetic acid than 20:3:1:0.4 for developing solvent, launch, take out, dry, put and inspect under 365nm ultra-violet lamp, in test sample chromatograph, with on reference substance chromatograph relevant position, the fluorescence speckle of aobvious same color.
Experimental example 5: the identification experiment of Rhizoma Kaempferiae
(1) reagent and test sample:
Reference substance: Ethyl p-methoxy-silicate reference substance (China pharmaceutical biological product calibrating academy), lot number: 0835-9601.
Test sample: SHEXIANG ZHUANGGU GAO (Shandong a person of good sense Fu Ruida hygienic material company limited), lot number: 20151105,20151106,20151107, specification is 7cm × 10cm.
(2) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 140cm2, adding volume ratio 40%～50% methanol 50mL, supersound process 25min, filter, filtrate is evaporated, and residue adds methanol 2mL and dissolves, as need testing solution；
(3) preparation of reference substance: take Ethyl p-methoxy-silicate reference substance, adds methanol and makes every 1mL solution containing 5mg, as reference substance solution；
(4) algoscopy: drawing need testing solution 6 μ L, reference substance solution 3 μ L puts respectively in same silica gel G F254On lamellae, with volume parts than 9:1 n-hexane-ethyl acetate for developing solvent, launch, take out, dry, put and inspect under 254nm ultra-violet lamp, in test sample chromatograph, on position corresponding with reference substance chromatograph, the speckle of aobvious same color.
The method have the advantages that
The invention provides the detection method of a kind of SHEXIANG ZHUANGGU GAO, including the discrimination method of Rhizoma Atractylodis, Herba Ephedrae, Radix Aconiti, Radix Aconiti Kusnezoffii, Rhizoma Zingiberis and/or Rhizoma Kaempferiae.In discriminating, test sample preparation is organic solvent supersound extraction, and method is easy, does not need backflow, reduces the application of organic solvent, stops the use of easily system poison reagent, reduces environmental pollution and the harm to laboratory personnel.This discrimination method favorable reproducibility, specificity are strong, can be used for the quality control of SHEXIANG ZHUANGGU GAO.
Detailed description of the invention
Following embodiment is used for further illustrating but is not limited to the present invention, and SHEXIANG ZHUANGGU GAO used is prepared by Shandong person of good sense Fu Ruida hygienic material company limited.
Embodiment 1: the detection method of a kind of Chinese medicine SHEXIANG ZHUANGGU GAO, including one or more in following discrimination method:
A. the discriminating of Rhizoma Atractylodis
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 210cm2Shredding and put in 100mL conical flask, add the 30mL volume ratio 40% ultrasonic 25min of methanol, filter, filtrate is evaporated, and residue adds dehydrated alcohol 2mL and dissolves, and obtains need testing solution；
(2) preparation of control medicinal material solution: take Rhizoma Atractylodis control medicinal material 0.8g, puts in 100mL conical flask, adds the 30mL volume ratio 40% ultrasonic 25min of methanol, filters, and filtrate is evaporated, and residue adds dehydrated alcohol 2mL and dissolves, and obtains control medicinal material solution；
(3) preparation of reference substance solution: take atisine chloride atractydin reference substance and add methanol and make every 1mL solution containing 0.2mg, as reference substance solution；
(4) algoscopy: draw need testing solution 20 μ L, control medicinal material solution 6 μ L, reference substance solution 2 μ L, put respectively on same silica gel g thin-layer plate, with the 60～90 of volume ratio 9:2 DEG C of petroleum ether-acetone for developing solvent, launch, take out, dry, spraying the ethanol solution of sulfuric acid with volume fraction 10%, it is clear to spot development to heat；In test sample chromatograph, on position corresponding with control medicinal material chromatograph and reference substance chromatograph, the speckle of aobvious same color；
B. the discriminating of Herba Ephedrae
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 210cm2Being cut into slice, add volume ratio 40% methanol 30mL, ultrasonic 30min, filter, filtrate is evaporated, and residue adds methanol 2mL and dissolves, as need testing solution；
(2) preparation of reference substance: take ephedrine hydrochloride reference substance, adds methanol and makes every lmL solution containing lmg, as reference substance solution；
(3) algoscopy: draw each 10 μ L of above two solution, put respectively on same silica gel g thin-layer plate, with the methylene chloride-methanol-ammonia of volume ratio 20:5:0.5 for developing solvent, launch, take out, dry, spray is with ninhydrin solution, clear to spot development 105 DEG C of heating；In test sample chromatograph, on position corresponding with reference substance chromatograph, aobvious identical punctation.
C. the discriminating of Radix Aconiti, Radix Aconiti Kusnezoffii
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 210cm2, shredding, put in conical flask, add volume ratio 40% methanol 40mL, supersound process 30min, filter, filtrate is evaporated, and residue adds isopropanol 2mL and dissolves, as need testing solution；
(2) preparation of reference substance: take aconitine reference substance and add isopropanol, makes every 1mL solution containing 1mg, as reference substance solution；
(3) algoscopy: draw each 5 μ L of above two solution, put respectively on same silica gel thin-layer plate, with volume parts than 6.4:3.6:1 n-hexane-ethyl acetate-methanol for developing solvent, put in the expansion cylinder of ammonia saturated with vapor 20min and launch, taking out, dry, spray is with rare bismuth potassium iodide test solution；In test sample chromatograph, with on reference substance chromatograph relevant position, the speckle of aobvious same color；
D. the discriminating of Rhizoma Zingiberis
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 210cm2It is cut into slice, adds volume ratio 40% methanol 40mL, supersound process 40min, let cool, filter, filtrate is evaporated, residue add water 20mL dissolve, with n-butyl alcohol jolting extract 2 times, each 30mL, merging n-butanol extracting liquid, be evaporated, residue adds 2mL methanol and dissolves, and obtains need testing solution；
(2) preparation of reference substance: take Rhizoma Zingiberis control medicinal material 1g, adds volume ratio 40% methanol 40mL, supersound process 40min, let cool, filter, filtrate is evaporated, residue add water 20mL dissolve, extract 2 times with n-butyl alcohol jolting, each 30mL, merges n-butanol extracting liquid, is evaporated, residue adds 2mL methanol and dissolves, and obtains control medicinal material solution；
(3) algoscopy: draw need testing solution 10 μ L, control medicinal material solution 5 μ L, put respectively on same silica gel g thin-layer plate, with the volume parts 60～90 DEG C of petroleum ether-ethyl acetate-methanol-glacial acetic acid than 20:3:1:0.4 for developing solvent, launch, take out, dry, put and inspect under 365nm ultra-violet lamp, in test sample chromatograph, with on reference substance chromatograph relevant position, the fluorescence speckle of aobvious same color；
E. the discriminating of Rhizoma Kaempferiae
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 210cm2, adding volume ratio 40% methanol 30mL, supersound process 30min, filter, filtrate is evaporated, and residue adds methanol 2mL and dissolves, as need testing solution；
(2) preparation of reference substance: take Ethyl p-methoxy-silicate reference substance, adds methanol and makes every 1mL solution containing 5mg, as reference substance solution；
(3) algoscopy: drawing need testing solution 6 μ L, reference substance solution 3 μ L puts respectively in same silica gel G F254On lamellae, with the volume parts n-hexane-ethyl acetate than 9:1 for developing solvent, launch, take out, dry, put and inspect under 254nm ultra-violet lamp, in test sample chromatograph, on position corresponding with reference substance chromatograph, the speckle of aobvious same color.
Embodiment 2: the detection method of a kind of Chinese medicine SHEXIANG ZHUANGGU GAO, including one or more in following discrimination method:
A. the discriminating of Rhizoma Atractylodis
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 210cm2, shredding, put in conical flask, add the 30mL volume ratio 40% ultrasonic 30min of methanol, filter, filtrate is evaporated, and residue adds dehydrated alcohol 2mL and dissolves, and obtains need testing solution；
(2) preparation of control medicinal material solution: take Rhizoma Atractylodis control medicinal material 0.8g, put in conical flask, adds the 30mL volume ratio 40% ultrasonic 30min of methanol, filters, and filtrate is evaporated, and residue adds dehydrated alcohol 2mL and dissolves, and obtains control medicinal material solution；
(3) preparation of reference substance solution: take atisine chloride atractydin reference substance and add methanol and make every 1mL solution containing 0.2mg, as reference substance solution；
(4) algoscopy: draw need testing solution 20 μ L, control medicinal material solution 6 μ L and reference substance solution 2 μ L, put respectively on same silica gel g thin-layer plate, with volume ratio be 9:2 60～90 DEG C of petroleum ether-acetone for developing solvent, launch, take out, drying, the ethanol solution of sulfuric acid that spray is 10% with volume ratio, it is clear to spot development to heat；In test sample chromatograph, on position corresponding with control medicinal material chromatograph and reference substance chromatograph, the speckle of aobvious same color；
B. the discriminating of Rhizoma Zingiberis
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 210cm2It is cut into slice, adds volume ratio 50% methanol 40mL, supersound process 30min, let cool, filter, filtrate is evaporated, residue add water 20mL dissolve, with n-butyl alcohol jolting extract 2 times, each 20mL, merging n-butanol extracting liquid, be evaporated, residue adds 2mL methanol and dissolves, and obtains need testing solution；
(2) preparation of reference substance: take Rhizoma Zingiberis control medicinal material 1g, adds volume ratio 50% methanol 40mL, supersound process 30min, let cool, filter, filtrate is evaporated, residue add water 20mL dissolve, extract 2 times with n-butyl alcohol jolting, each 20mL, merges n-butanol extracting liquid, is evaporated, residue adds 2mL methanol and dissolves, and obtains control medicinal material solution；
(3) algoscopy: draw need testing solution 10 μ L, control medicinal material solution 5 μ L, put respectively on same silica gel g thin-layer plate, with the volume parts 60～90 DEG C of petroleum ether-ethyl acetate-methanol-glacial acetic acid than 20:3:1:0.5 for developing solvent, launch, take out, dry, put and inspect under 365nm ultra-violet lamp, in test sample chromatograph, with on reference substance chromatograph relevant position, the fluorescence speckle of aobvious same color；
C. the discriminating of Rhizoma Kaempferiae
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 210cm2, adding methanol 40mL, supersound process 30min, filter, filtrate is evaporated, and residue adds methanol 2mL and dissolves, as need testing solution；
(2) preparation of reference substance: take Ethyl p-methoxy-silicate reference substance, adds methanol and makes every 1mL solution containing 5mg, as reference substance solution；
(3) algoscopy: drawing need testing solution 6 μ L, reference substance solution 3 μ L puts respectively in same silica gel G F254On lamellae, with volume parts than 9:1 n-hexane-ethyl acetate for developing solvent, launch, take out, dry, put and inspect under 254nm ultra-violet lamp, in test sample chromatograph, on position corresponding with reference substance chromatograph, the speckle of aobvious same color.
Embodiment 3: the detection method of a kind of Chinese medicine SHEXIANG ZHUANGGU GAO, including one or more in following discrimination method:
A. the discriminating of Rhizoma Atractylodis
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 140cm2, shredding, put in conical flask, add the ultrasonic 30min of 25mL methanol, filter, filtrate is evaporated, and residue adds dehydrated alcohol 1mL and dissolves, and obtains need testing solution；
(2) preparation of reference substance: take Rhizoma Atractylodis control medicinal material 0.8g, put in conical flask, adds the ultrasonic 30min of 25mL methanol, filters, and filtrate is evaporated, and residue adds dehydrated alcohol 2mL and dissolves, and obtains control medicinal material solution；Take atisine chloride atractydin reference substance again to add methanol and make every 1mL solution containing 0.2mg, as reference substance solution；
(3) algoscopy: draw need testing solution 10 μ L, control medicinal material solution 6 μ L and reference substance solution 2 μ L, put respectively on same silica gel g thin-layer plate, with volume ratio be 9:1 60～90 DEG C of petroleum ether-acetone for developing solvent, launch, take out, drying, the ethanol solution of sulfuric acid that spray is 10% with volume ratio, it is clear to spot development to heat；In test sample chromatograph, on position corresponding with control medicinal material chromatograph and reference substance chromatograph, the speckle of aobvious same color；
B. the discriminating of Herba Ephedrae
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 140cm2, it being cut into slice, add volume ratio 60% methanol 30mL, supersound process 25min, filter, filtrate is evaporated, and residue adds methanol 2mL and dissolves, and filtrate is as need testing solution；
(2) preparation of reference substance: take ephedrine hydrochloride reference substance, adds methanol and makes every lmL solution containing lmg, as reference substance solution；
(3) algoscopy: draw each 12 μ L of above two solution, put respectively on same silica gel g thin-layer plate, with the methylene chloride-methanol-ammonia of volume ratio 20:5:0.5 for developing solvent, launch, take out, dry, spray is with ninhydrin solution, clear to spot development 105 DEG C of heating；In test sample chromatograph, on position corresponding with reference substance chromatograph, aobvious identical punctation.
C. the discriminating of Radix Aconiti, Radix Aconiti Kusnezoffii
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 210cm2, shredding, put in conical flask, add isopropanol 40mL, supersound process 30min, filter, filtrate is evaporated, and residue adds isopropanol 2mL and dissolves, as need testing solution；
(2) preparation of reference substance: take aconitine reference substance and add aqueous isopropanol, makes every 1mL solution containing 1mg, as reference substance solution；
(3) algoscopy: draw each 5 μ L of above two solution, put respectively on same silica gel thin-layer plate, with volume parts than 6.4:3.6:1 n-hexane-ethyl acetate-methanol for developing solvent, put in the expansion cylinder of ammonia saturated with vapor 20min and launch, taking out, dry, spray is with rare bismuth potassium iodide test solution；In test sample chromatograph, with on reference substance chromatograph relevant position, the speckle of aobvious same color；
D. the discriminating of Rhizoma Zingiberis
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 210cm2Be cut into slice, add methanol 40mL, supersound process 40min, let cool, filter, filtrate is evaporated, residue add water 20mL dissolve, with n-butyl alcohol jolting extract 2 times, each 30mL, merge n-butanol extracting liquid, be evaporated, residue add 2mL methanol dissolve, obtain need testing solution；
(2) preparation of reference substance: take Rhizoma Zingiberis control medicinal material 1g, add methanol 40mL, supersound process 40min, let cool, filter, filtrate is evaporated, residue add water 20mL dissolve, extract 2 times with n-butyl alcohol jolting, each 30mL, merges n-butanol extracting liquid, is evaporated, residue adds 2mL methanol and dissolves, and obtains control medicinal material solution；
(3) algoscopy: draw need testing solution 10 μ L, control medicinal material solution 5 μ L, put respectively on same silica gel g thin-layer plate, with the volume parts 60～90 DEG C of petroleum ether-ethyl acetate-methanol-glacial acetic acid than 19:3:1:0.5 for developing solvent, launch, take out, dry, put and inspect under 365nm ultra-violet lamp, in test sample chromatograph, with on reference substance chromatograph relevant position, the fluorescence speckle of aobvious same color.
Embodiment 4: the detection method of a kind of Chinese medicine SHEXIANG ZHUANGGU GAO, including following discrimination method:
A. the discriminating of Radix Aconiti, Radix Aconiti Kusnezoffii
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 140cm2Sheet, shreds, and puts in conical flask, adds isopropanol 25mL, supersound process 30min, and filtered solution is concentrated into 1mL, as need testing solution；
(2) preparation of reference substance: take aconitine reference substance and add aqueous isopropanol, makes every 1mL solution containing 1mg, as reference substance solution；
(3) algoscopy: draw each 5 μ L of above two solution, put respectively on same silica gel thin-layer plate, with volume parts than 6.4:3.6:1 n-hexane-ethyl acetate-methanol for developing solvent, put in the expansion cylinder of ammonia saturated with vapor 20min and launch, taking out, dry, spray is with rare bismuth potassium iodide test solution；In test sample chromatograph, with on reference substance chromatograph relevant position, the speckle of aobvious same color.
Claims (8)
1. the detection method of a SHEXIANG ZHUANGGU GAO, it is characterised in that include following discrimination method:
A. the discriminating of Rhizoma Atractylodis
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 2～5, shred, put in conical flask, adding volume ratio 40%～50% ultrasonic 15～30min of methanol 25～50mL, filter, filtrate is evaporated, residue adds dehydrated alcohol 2mL and dissolves, and obtains need testing solution；
(2) preparation of reference substance: take Rhizoma Atractylodis control medicinal material 0.8g, put in conical flask, adds 25～50mL volume ratio 40%～50% ultrasonic 15～30min of methanol, filters, and filtrate is evaporated, and residue adds dehydrated alcohol 2mL and dissolves, and obtains control medicinal material solution；Take atisine chloride atractydin reference substance again to add methanol and make every 1mL solution containing 0.2～0.5mg, as reference substance solution；
(3) algoscopy: draw need testing solution, control medicinal material solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with volume ratio be 8～9:1～2 60～90 DEG C of petroleum ether-acetone for developing solvent, launch, take out, drying, the ethanol solution of sulfuric acid that spray is 10% with volume ratio, it is clear to spot development to heat；In test sample chromatograph, on position corresponding with control medicinal material chromatograph and reference substance chromatograph, the speckle of aobvious same color；
B. the discriminating of Herba Ephedrae
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 2～5 and be cut into slice, adds the methanol 20～50mL of volume ratio 40%～50%, supersound process 20～30min, filters, and filtrate is evaporated, and residue adds methanol 2mL and dissolves, as need testing solution；
(2) preparation of reference substance: take ephedrine hydrochloride reference substance, adds methanol and makes every lmL solution containing lmg, as reference substance solution；
(3) algoscopy: draw each 10～15 μ L of above two solution, put respectively on same silica gel g thin-layer plate, with the methylene chloride-methanol-ammonia of volume ratio 15～20:2～6:0.5～1 for developing solvent, launch, take out, drying, spray is with ninhydrin solution, clear to spot development 105 DEG C of heating；In test sample chromatograph, on position corresponding with reference substance chromatograph, the speckle of aobvious same color；
C. the discriminating of Radix Aconiti, Radix Aconiti Kusnezoffii
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 2～5, shred, put in conical flask, add the organic solvent 25～50mL of volume ratio 40%～50%, supersound process 20～30min, filter, filtrate is evaporated, and residue adds organic solvent 2mL and dissolves, as need testing solution；
(2) preparation of reference substance: take aconitine reference substance and add organic solvent, makes every 1mL solution containing 0.5～1mg, as reference substance solution；
(3) algoscopy: draw each 5 μ L of above two solution, put respectively on same silica gel g thin-layer plate, with volume parts n-hexane-ethyl acetate-methanol than 6～7:3～4:1 for developing solvent, put in the expansion cylinder of ammonia saturated with vapor 20～30min and launch, take out, drying, spray is with rare bismuth potassium iodide test solution；In test sample chromatograph, with on reference substance chromatograph relevant position, the speckle of aobvious same color；
D. the discriminating of Rhizoma Zingiberis
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 2～5 and be cut into slice, add volume ratio 40%～50% methanol 20～50mL, supersound process 20～40min, let cool, filter, filtrate is evaporated, residue add water 20mL dissolve, extract 2～3 times with n-butyl alcohol jolting, each 30mL, merges n-butanol extracting liquid, is evaporated, residue adds 2～dissolving of 3mL methanol, obtains need testing solution；
(2) preparation of reference substance: take Rhizoma Zingiberis control medicinal material 1g, adds volume ratio 40%～50% methanol 20～50mL, supersound process 20～40min, let cool, filter, filtrate is evaporated, residue add water 20mL dissolve, extract 2～3 times with n-butyl alcohol jolting, each 30mL, merges n-butanol extracting liquid, is evaporated, residue adds 2～dissolving of 3mL methanol, obtains control medicinal material solution；
(3) algoscopy: draw need testing solution 10 μ L, control medicinal material solution 5 μ L, put respectively on same silica gel g thin-layer plate, with volume parts than 60～90 DEG C of petroleum ether-ethyl acetate-methanol-glacial acetic acid of 18～20:1～3:0.5～1:0～1 for developing solvent, launch, take out, dry, put and inspect under 365nm ultra-violet lamp, in test sample chromatograph, with on reference substance chromatograph relevant position, the fluorescence speckle of aobvious same color；
E. the discriminating of Rhizoma Kaempferiae
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 2～5, adds volume ratio 40%～50% methanol 25～50mL, supersound process 15～30min, filters, and filtrate is evaporated, and residue adds methanol 2mL and dissolves, as need testing solution；
(2) preparation of reference substance: take Ethyl p-methoxy-silicate reference substance, adds methanol and makes every 1mL solution containing 1～5mg, as reference substance solution；
(3) algoscopy: drawing need testing solution 6 μ L, reference substance solution 3 μ L puts respectively in same silica gel G F254On lamellae, with volume parts than 8～9:1～2 n-hexane-ethyl acetate for developing solvent, launch, take out, dry, put and inspect under 254nm ultra-violet lamp, in test sample chromatograph, on position corresponding with reference substance chromatograph, the speckle of aobvious same color.
2. the detection method of Chinese medicine SHEXIANG ZHUANGGU GAO according to claim 1, it is characterised in that in the discriminating of described Rhizoma Atractylodis, developing solvent is the volume parts 60～90 DEG C of petroleum ether-acetone than 9:2.
3. the detection method of SHEXIANG ZHUANGGU GAO according to claim 1, it is characterised in that in the discriminating of described Herba Ephedrae, solvent for use is the methanol of volume ratio 40%.
4. the detection method of Chinese medicine SHEXIANG ZHUANGGU GAO according to claim 1, it is characterised in that in the discriminating of described Herba Ephedrae, developing solvent is volume parts methylene chloride-methanol-ammonia than 20:5:0.5.
5. the detection method of Chinese medicine SHEXIANG ZHUANGGU GAO according to claim 1, it is characterized in that, in described Radix Aconiti, the discriminating of Radix Aconiti Kusnezoffii, organic reagent can be isopropanol, chloroform, methanol, and developing solvent is volume parts n-hexane-ethyl acetate-methanol than 6.4:3.6:1.
6. the detection method of Chinese medicine SHEXIANG ZHUANGGU GAO according to claim 1, it is characterised in that in the discriminating of described Rhizoma Zingiberis, developing solvent is the volume parts 60～90 DEG C of petroleum ether-ethyl acetate-methanol-glacial acetic acid than 20:3:1:0.4.
7. the detection method of Chinese medicine SHEXIANG ZHUANGGU GAO according to claim 1, it is characterised in that in the discriminating of described Rhizoma Kaempferiae, developing solvent is the volume parts n-hexane-ethyl acetate than 9:1.
8. the detection method of SHEXIANG ZHUANGGU GAO according to claim 1, it is characterised in that include following discrimination method:
A. the discriminating of Rhizoma Atractylodis
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 210cm2Shredding and put in 100mL conical flask, add the 30mL volume ratio 40% ultrasonic 25min of methanol, filter, filtrate is evaporated, and residue adds dehydrated alcohol 2mL and dissolves, and obtains need testing solution；
(2) preparation of reference substance: take Rhizoma Atractylodis control medicinal material 0.8g, puts in 100mL conical flask, adds the 30mL volume ratio 40% ultrasonic 25min of methanol, filters, and filtrate is evaporated, and residue adds dehydrated alcohol 2mL and dissolves, and obtains control medicinal material solution；Take atisine chloride atractydin reference substance again to add methanol and make every 1mL solution containing 0.2mg, as reference substance solution；
(3) algoscopy: draw need testing solution 20 μ L, control medicinal material solution 6 μ L, reference substance solution 2 μ L, put respectively on same silica gel g thin-layer plate, with the 60～90 of volume ratio 9:2 DEG C of petroleum ether-acetone for developing solvent, launch, take out, dry, spraying the ethanol solution of sulfuric acid with volume fraction 10%, it is clear to spot development to heat；In test sample chromatograph, on position corresponding with control medicinal material chromatograph and reference substance chromatograph, the speckle of aobvious same color；
B. the discriminating of Herba Ephedrae
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 210cm2Being cut into slice, add volume ratio 40% methanol 30mL, ultrasonic 30min, filter, filtrate is evaporated, and residue adds methanol 2mL and dissolves, as need testing solution；
(2) preparation of reference substance: take ephedrine hydrochloride reference substance, adds methanol and makes every lmL solution containing lmg, as reference substance solution；
(3) algoscopy: draw each 10 μ L of above two solution, put respectively on same silica gel g thin-layer plate, with the methylene chloride-methanol-ammonia of volume ratio 20:5:0.5 for developing solvent, launch, take out, dry, spray is with ninhydrin solution, clear to spot development 105 DEG C of heating；In test sample chromatograph, on position corresponding with reference substance chromatograph, aobvious identical punctation；
C. the discriminating of Radix Aconiti, Radix Aconiti Kusnezoffii
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 210cm2, shredding, put in conical flask, add volume ratio 40% methanol 40mL, supersound process 30min, filter, filtrate is evaporated, and residue adds isopropanol 2mL and dissolves, as need testing solution；
(2) preparation of reference substance: take aconitine reference substance and add isopropanol, makes every 1mL solution containing 1mg, as reference substance solution；
(3) algoscopy: draw each 5 μ L of above two solution, put respectively on same silica gel thin-layer plate, with volume parts than 6.4:3.6:1 n-hexane-ethyl acetate-methanol for developing solvent, put in the expansion cylinder of ammonia saturated with vapor 20min and launch, taking out, dry, spray is with rare bismuth potassium iodide test solution；In test sample chromatograph, with on reference substance chromatograph relevant position, the speckle of aobvious same color；
D. the discriminating of Rhizoma Zingiberis
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 210cm2It is cut into slice, adds volume ratio 40% methanol 40mL, supersound process 40min, let cool, filter, filtrate is evaporated, residue add water 20mL dissolve, with n-butyl alcohol jolting extract 2 times, each 30mL, merging n-butanol extracting liquid, be evaporated, residue adds 2mL methanol and dissolves, and obtains need testing solution；
(2) preparation of reference substance: take Rhizoma Zingiberis control medicinal material 1g, adds volume ratio 40% methanol 40mL, supersound process 40min, let cool, filter, filtrate is evaporated, residue add water 20mL dissolve, extract 2 times with n-butyl alcohol jolting, each 30mL, merges n-butanol extracting liquid, is evaporated, residue adds 2mL methanol and dissolves, and obtains control medicinal material solution；
(3) algoscopy: draw need testing solution 10 μ L, control medicinal material solution 5 μ L, put respectively on same silica gel g thin-layer plate, with the volume parts 60～90 DEG C of petroleum ether-ethyl acetate-methanol-glacial acetic acid than 20:3:1:0.4 for developing solvent, launch, take out, dry, put and inspect under 365nm ultra-violet lamp, in test sample chromatograph, with on reference substance chromatograph relevant position, the fluorescence speckle of aobvious same color；
E. the discriminating of Rhizoma Kaempferiae
According to Chinese Pharmacopoeia four general rule 0502 tlc determinations of version in 2015；
(1) preparation of need testing solution: take SHEXIANG ZHUANGGU GAO 210cm2, adding volume ratio 40% methanol 30mL, supersound process 30min, filter, filtrate is evaporated, and residue adds methanol 2mL and dissolves, as need testing solution；
(2) preparation of reference substance: take Ethyl p-methoxy-silicate reference substance, adds methanol and makes every 1mL solution containing 5mg, as reference substance solution；
(3) algoscopy: drawing need testing solution 6 μ L, reference substance solution 3 μ L puts respectively in same silica gel G F254On lamellae, with the volume parts n-hexane-ethyl acetate than 9:1 for developing solvent, launch, take out, dry, put and inspect under 254nm ultra-violet lamp, in test sample chromatograph, on position corresponding with reference substance chromatograph, the speckle of aobvious same color.
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| CN106546691A (en) * | 2016-10-20 | 2017-03-29 | 中悦民安（北京）科技发展有限公司 | The discrimination method of Rhizoma Atractylodis in Chinese medicine compound |
| CN107167543A (en) * | 2017-06-20 | 2017-09-15 | 安徽安科余良卿药业有限公司 | The structure and its quality determining method of SHEXIANG ZHUANGGU GAO standard feature collection of illustrative plates |
| CN113588852A (en) * | 2021-08-16 | 2021-11-02 | 浙江鼎泰药业股份有限公司 | Method for measuring content of volatile components in high-activity transdermal plaster |
| CN113740198A (en) * | 2021-08-17 | 2021-12-03 | 马应龙药业集团股份有限公司 | Method for determining yellow vaseline content in Mayinglong musk hemorrhoid ointment |
| CN115260032A (en) * | 2022-10-08 | 2022-11-01 | 中国科学院昆明植物研究所 | Method for separating and purifying ethyl p-methoxycinnamate and/or ethyl cinnamate by high-speed counter-current chromatography |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2732259B2 (en) * | 1988-09-10 | 1998-03-25 | 三和生薬株式会社 | New aconitine compounds and analgesic / anti-inflammatory agents |
| US20030026850A1 (en) * | 2001-07-13 | 2003-02-06 | Bottaro Jeffrey C. | N,N-dinitramide salts as solubilizing agents for biologically active agents |
| CN102429966A (en) * | 2011-01-14 | 2012-05-02 | 成都中医药大学 | Gall-free Heishunpian or rhizoma typhonii tablet and processing method thereof |
| CN102507781A (en) * | 2011-11-09 | 2012-06-20 | 云南金乌黑药制药有限公司 | Method for controlling quality of medicine for treating rheumatism and apoplexy diseases |
| CN103784664A (en) * | 2014-01-27 | 2014-05-14 | 贵州师范大学 | Dual-phase capsule for preventing and treating chronic pelvic inflammation and preparation method and detection method thereof |
| CN104949955A (en) * | 2014-06-26 | 2015-09-30 | 中国人民解放军第二军医大学 | Method for detecting ephedrine additives doped in weight-losing traditional Chinese medicines |
-
2016
- 2016-04-18 CN CN201610240222.XA patent/CN105784914A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2732259B2 (en) * | 1988-09-10 | 1998-03-25 | 三和生薬株式会社 | New aconitine compounds and analgesic / anti-inflammatory agents |
| US20030026850A1 (en) * | 2001-07-13 | 2003-02-06 | Bottaro Jeffrey C. | N,N-dinitramide salts as solubilizing agents for biologically active agents |
| CN102429966A (en) * | 2011-01-14 | 2012-05-02 | 成都中医药大学 | Gall-free Heishunpian or rhizoma typhonii tablet and processing method thereof |
| CN102507781A (en) * | 2011-11-09 | 2012-06-20 | 云南金乌黑药制药有限公司 | Method for controlling quality of medicine for treating rheumatism and apoplexy diseases |
| CN103784664A (en) * | 2014-01-27 | 2014-05-14 | 贵州师范大学 | Dual-phase capsule for preventing and treating chronic pelvic inflammation and preparation method and detection method thereof |
| CN104949955A (en) * | 2014-06-26 | 2015-09-30 | 中国人民解放军第二军医大学 | Method for detecting ephedrine additives doped in weight-losing traditional Chinese medicines |
Non-Patent Citations (9)
| Title |
|---|
| 刘松青 等: "必克敏胶囊质量标准的研究", 《中国药房》 * |
| 国家药典委员会 编: "《中华人民共和国药典2015年版一部》", 30 June 2015 * |
| 国家药品监督管理局 发布: "《国家药品监督局标准（试行）WS-11263（ZD-1263）-2002》", 1 December 2002 * |
| 徐小芳 等: "复方附子口服液中黄芪甲苷含量测定和乌头碱限量检查方法研究", 《中南药学》 * |
| 成差群 等: "追风活血膏不同制法化学等量性研究", 《中药材》 * |
| 李萌 等: "LC-MS/MS测定乌头属中药中8个生物碱含量", 《辽宁中医药大学学报》 * |
| 柳俊: "麝香壮骨膏中药材浸膏的提取工艺及质量标准研究", 《中国优秀硕士学位论文全文数据库 工程科技Ⅰ辑》 * |
| 胡丹 等: "新型狗皮膏中的鉴别试验", 《中国医药指南》 * |
| 蔡博 等: "超高效液相色谱质谱联用技术同时测定小金丸中 7 种成分的含量", 《中国药学杂志》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106546691A (en) * | 2016-10-20 | 2017-03-29 | 中悦民安（北京）科技发展有限公司 | The discrimination method of Rhizoma Atractylodis in Chinese medicine compound |
| CN107167543A (en) * | 2017-06-20 | 2017-09-15 | 安徽安科余良卿药业有限公司 | The structure and its quality determining method of SHEXIANG ZHUANGGU GAO standard feature collection of illustrative plates |
| CN113588852A (en) * | 2021-08-16 | 2021-11-02 | 浙江鼎泰药业股份有限公司 | Method for measuring content of volatile components in high-activity transdermal plaster |
| CN113740198A (en) * | 2021-08-17 | 2021-12-03 | 马应龙药业集团股份有限公司 | Method for determining yellow vaseline content in Mayinglong musk hemorrhoid ointment |
| CN113740198B (en) * | 2021-08-17 | 2024-03-08 | 马应龙药业集团股份有限公司 | Method for measuring yellow vaseline content in musk hemorrhoid ointment |
| CN115260032A (en) * | 2022-10-08 | 2022-11-01 | 中国科学院昆明植物研究所 | Method for separating and purifying ethyl p-methoxycinnamate and/or ethyl cinnamate by high-speed counter-current chromatography |
| CN115260032B (en) * | 2022-10-08 | 2023-01-06 | 中国科学院昆明植物研究所 | Method for separating and purifying ethyl p-methoxycinnamate and/or ethyl cinnamate by high-speed counter-current chromatography |
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