CN102091175A - Method for identifying traditional Chinese medicinal preparation for treatment or adjuvant treatment of tumor diseases - Google Patents
Method for identifying traditional Chinese medicinal preparation for treatment or adjuvant treatment of tumor diseases Download PDFInfo
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Abstract
The invention relates to a method for identifying a traditional Chinese medicinal preparation for the treatment or adjuvant treatment of tumor diseases, and belongs to the technical field of the quality control of traditional Chinese medicinal preparations. The traditional Chinese medicinal preparation of the invention is a newly developed Chinese medicinal compound preparation, and in order to control the quality of the preparation, a method for identifying placenta hominis, prepared rehmannia root, glossy privet fruit and mongolian snakegourd root in the traditional Chinese medicinal preparation by thin layer chromatography is established. The method has high specificity, repeatability and durability, and can be used for identifying corresponding medicines in the traditional Chinese medicinal preparation.
Description
Technical field
The present invention relates to a kind of discrimination method of Chinese medicine preparation, particularly relate to the discrimination method of the Chinese medicine preparation of a kind of treatment or adjuvant therapy of tumors disease, belong to Chinese medicine preparation Quality Control Technology field.
Background technology
The Chinese medicine preparation of a kind of treatment of the present invention or adjuvant therapy of tumors disease is the compound Chinese medicinal preparation that we newly develop, be mainly used in the treatment or the auxiliary treatment of QIXUELIANGXU type tumor disease, or be used for that tumor is put, deficiency of both QI and blood that chemotherapy and postoperative body void cause.In order to control this Chinese medicine preparation quality, we have carried out Study on Identification to this Chinese medicine preparation.
The discriminating of Chinese medicine preparation always is exactly a difficult point, have the saying of " pill, powder, extract and pellet, angle's difficulty are distinguished ".At present, the discrimination method of Chinese medicine preparation mainly contains: microscopical identification method, general physicochemical identification method (as: an appendix IV of Chinese Pharmacopoeia version in 2010 general identification test), ultraviolet visible spectrophotometry, paper chromatography, thin layer chromatography, high performance liquid chromatography, gas chromatography etc.; In addition, also has finger printing discriminating etc.These discrimination methods are each has something to recommend him, should select which type of discrimination method to a Chinese medicine preparation actually, need repetition test research and methodology to verify and select.If selected wherein a kind of discrimination method for use, as thin layer chromatography, need its used immobile phase of research, contrast material etc., need research developing solvent, developer, also need to study preparation method of need testing solution and contrast substance solution or the like, comprised need testing solution preparation technology, contrast material and preparation technology thereof, immobile phase, developing solvent, point sample amount, launch temperature, coloration method, inspection method etc.Just because of this, the science of these discrimination methods has progressively obtained the approval of international community, and the modernization of Chinese medicine preparation quality standard begins progressively to realize.
Thin layer chromatography is called for short the TLC method, be with the need testing solution point on lamellae, in launching container, launch with developing solvent, the test sample ingredient is separated, gained chromatogram and the suitable contrast material chromatogram contrast of gained by the same method are used for differentiating.Thin layer chromatography mainly partly is made of immobile phase, developing solvent, developer etc., is made of instruments such as lamellae, sample applicator, expansion container, display device, checking device and material.The most frequently used immobile phase of lamellae has silica gel G, silica gel G F
254, silica gel H, silica gel H F
254, microcrystalline Cellulose etc.
The specificity of discrimination method has directly determined the value of this discrimination method, if specificity is not strong, false positive takes place easily.At present, the discrimination method specificity of many Chinese medicine preparation is not strong, mixes the tripolycyanamide incident in the milk powder that China once took place, and fails in time to test out, and is exactly because discrimination method specificity at that time is not strong, has produced false positive.Placenta Hominis, Radix Rehmanniae Preparata, Fructus Ligustri Lucidi, Radix Trichosanthis are the contained flavour of a drug of preparation of the present invention, and discrimination method provided by the invention has very strong specificity.
Summary of the invention
The discrimination method that the purpose of this invention is to provide the Chinese medicine preparation of a kind of treatment or adjuvant therapy of tumors disease, adopt thin layer chromatography to differentiate whether to contain in this Chinese medicine preparation in Placenta Hominis, Radix Rehmanniae Preparata, Fructus Ligustri Lucidi, the Radix Trichosanthis any or any two or wantonly three kinds or whole, this method specificity is strong, repeatability, ruggedness are good, and the corresponding flavour of a drug that can be used in this Chinese medicine preparation are differentiated.
The technical solution used in the present invention is as follows:
The discrimination method of the Chinese medicine preparation of a kind of treatment or adjuvant therapy of tumors disease, it is characterized in that: comprise four kinds of Placenta Hominiss, Radix Rehmanniae Preparata, Fructus Ligustri Lucidi, Radix Trichosanthis in the described Chinese medicine preparation, described discrimination method comprise following discrimination method any or any two or wantonly three kinds or all:
(1) discrimination method of Placenta Hominis in the described Chinese medicine preparation is to be the thin layer chromatography discrimination method of contrast with the Placenta Hominis control medicinal material, may further comprise the steps:
1. the preparation of need testing solution: get described Chinese medicine preparation, add chloroform or dichloromethane, supersound process filters, and filtrate concentrates, and makes per 1 mL and contains the solution that the crude drug amount is equivalent to Placenta Hominis 0.2~5g, as need testing solution;
2. the preparation of control medicinal material solution: get the Placenta Hominis control medicinal material, add chloroform or dichloromethane, supersound process filters, and filtrate concentrates, and makes per 1 mL and contains the solution that the crude drug amount is equivalent to Placenta Hominis 0.2~5g, in contrast medical material solution;
3. algoscopy: according to the test of an appendix VI of Chinese Pharmacopoeia version in 2010 B thin layer chromatography, drawing each 2~20 μ L of control medicinal material solution and need testing solution, put respectively on same silica gel g thin-layer plate, is developing solvent with normal hexane-ethyl acetate 9: 1 ~ 9, launch, take out, dry, spray is with 5%~50% perchloric acid solution, it is clear to be heated to speckle colour developing at 60~120 ℃, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the principal spot of same color;
(2) discrimination method of Radix Rehmanniae Preparata in the described Chinese medicine preparation is to be the thin layer chromatography discrimination method of contrast with the Radix Rehmanniae Preparata control medicinal material, may further comprise the steps:
1. the preparation of need testing solution: get described Chinese medicine preparation, add water, warmly make fully molten loosing, be heated to and boil, put cold, centrifugal, get supernatant, concentrate, extract 1~3 time combined ethyl acetate liquid with the ethyl acetate jolting, evaporate to dryness, residue adds methanol makes dissolving, makes per 1 mL and contains the solution that the crude drug amount is equivalent to Radix Rehmanniae Preparata 2~20g, as need testing solution;
2. the preparation of control medicinal material solution: get the Radix Rehmanniae Preparata control medicinal material, decoct with water 1~2 time, filter with absorbent cotton, merging filtrate concentrates, extract 1~3 time with the ethyl acetate jolting, combined ethyl acetate liquid, evaporate to dryness, residue add methanol makes dissolving, make per 1 mL and contain the solution that the crude drug amount is equivalent to Radix Rehmanniae Preparata 2~20g, in contrast medical material solution;
3. algoscopy: according to the test of an appendix VI of Chinese Pharmacopoeia version in 2010 B thin layer chromatography, draw each 2~20 μ L of control medicinal material solution and need testing solution, put respectively on same silica gel g thin-layer plate, with o-Dimethylbenzene-ethyl acetate 1:1 is developing solvent, wherein o-Dimethylbenzene is replaceable is benzene, or toluene, or meta-xylene, or xylol, or o-Dimethylbenzene, meta-xylene, three kinds of mixture of pressing arbitrary proportion of xylol launch, and take out, dry, spray is with 0. 1% 2,4 dinitrophenyl hydrazine alcoholic solution, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the principal spot of same color;
Wherein 0.1% 2,4 dinitrophenyl hydrazine alcoholic solution is: get 2,4 dinitrophenyl hydrazine 1g, add dehydrated alcohol 1000 mL and make dissolving, slowly add concentration again and be 36%~38% hydrochloric acid 10mL, shake up, promptly;
(3) discrimination method of Fructus Ligustri Lucidi in the described Chinese medicine preparation is to be the thin layer chromatography discrimination method of contrast with Fructus Ligustri Lucidi control medicinal material and oleanolic acid reference substance, may further comprise the steps:
1. the preparation of need testing solution: get described Chinese medicine preparation, add chloroform or dichloromethane, extract 1~3 time, merge chloroform liquid or dichloromethane solution, filter, the filtrate evaporate to dryness, residue adds methanol, and perhaps residue adds dehydrated alcohol-chloroform 3:2 mixed solution, at 5~50 ℃, make per 1 mL and contain the solution that the crude drug amount is equivalent to Fructus Ligustri Lucidi 0.2~5g, as need testing solution;
2. the preparation of control medicinal material solution: get the Fructus Ligustri Lucidi control medicinal material, add chloroform or dichloromethane, supersound process, filter, the filtrate evaporate to dryness, residue adds methanol, perhaps residue adds dehydrated alcohol-chloroform 3:2 mixed solution, at 5~50 ℃, make per 1 mL and contain the solution that the crude drug amount is equivalent to Fructus Ligustri Lucidi 0.2~5g, in contrast medical material solution;
3. the preparation of reference substance solution: even up pier fruit acid reference substance, add methanol or 95% ethanol, at 5~50 ℃, be mixed with the solution that every 1mL contains 1mg, in contrast product solution;
4. algoscopy: according to the test of an appendix VI of Chinese Pharmacopoeia version in 2010 B thin layer chromatography, at 5~50 ℃, draw need testing solution, each 2~20 μ L of control medicinal material solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with cyclohexane extraction-acetone-ethyl acetate-glacial acetic acid 7~15: 2~7: 1~5:0~0.5, be developing solvent perhaps with chloroform-methanol-formic acid 20~40:1:0.3~1.1, launch, take out, dry, spray is with 2%~20% ethanol solution of sulfuric acid, perhaps spray with 1%~10% phosphomolybdic acid ethanol solution, it is clear to be heated to speckle colour developing at 60~120 ℃, in the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the principal spot of same color;
If the content of Fructus Ligustri Lucidi is lower in the preparation, before the preparation need testing solution, can carry out pretreatment to said preparation, to reach better identification result, concrete operations are: get described Chinese medicine preparation, add 0.2%~0.5% sodium hydroxide solution, warmly make fully molten loosing, be heated to and boil, put cold, filtrate or supernatant are got in filtration or centrifugal, regulate pH value to 1~2 with hydrochloric acid or sulphuric acid; If Fructus Ligustri Lucidi content height in the preparation does not generally need to carry out pretreatment;
(4) discrimination method of Radix Trichosanthis in the described Chinese medicine preparation is to be the thin layer chromatography discrimination method of contrast with Radix Trichosanthis control medicinal material and Citruline reference substance, may further comprise the steps:
1. the preparation of need testing solution: get described Chinese medicine preparation, add 30%~100% ethanol, supersound process filters, the filtrate evaporate to dryness adds 50% ethanol and makes dissolving, filters, get filtrate, make per 1 mL and contain the solution that the crude drug amount is equivalent to Radix Trichosanthis 0.01~1g, as need testing solution;
2. the preparation of control medicinal material solution: get the Radix Trichosanthis control medicinal material, add 50% ethanol, supersound process filters, and gets filtrate, makes per 1 mL and contains the solution that the crude drug amount is equivalent to Radix Trichosanthis 0.01~1g, in contrast medical material solution;
3. the preparation of reference substance solution: get the Citruline reference substance, add 50% ethanol and be mixed with the solution that every 1mL contains 1mg, in contrast product solution;
4. algoscopy: according to the test of an appendix VI of Chinese Pharmacopoeia version in 2010 B thin layer chromatography, draw need testing solution, each 1~20 μ L of control medicinal material solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with n-butyl alcohol-dehydrated alcohol-glacial acetic acid-water 7~15: 0~2:2 ~ 6:1~5, n-butyl alcohol-formic acid-water 15~25: 3:2, n-butyl alcohol-acetone-water 2~5: any among the 2:1 is developing solvent, launch, take out, dry, spray is with 1%~10% ethanol solution of ninhydrin, it is clear to be heated to speckle colour developing at 60~120 ℃, in the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the principal spot of same color.
Described Chinese medicine preparation by Placenta Hominis, the Radix Astragali, Radix Rehmanniae Preparata, Radix Codonopsis, the Rhizoma Atractylodis Macrocephalae, Radix Angelicae Sinensis, Radix Glycyrrhizae, Fructus Ligustri Lucidi, Rhizoma Polygonati, Herba Gueldenstaedtiae or Herba Violae, Radix Trichosanthis ten simply Chinese medicine form, Placenta Hominis is ground into powder, all the other ten flavor medical material solvents extract, and make tablet, capsule, granule, powder, the various suitable dosage forms of pill.
Further, the prescription of described Chinese medicine preparation is counted by weight ratio: Placenta Hominis 4~7, the Radix Astragali 20~50, Radix Rehmanniae Preparata 10~30, Radix Codonopsis 5~20, the Rhizoma Atractylodis Macrocephalae 5~20, Radix Angelicae Sinensis 5~15, Radix Glycyrrhizae 5~15, Fructus Ligustri Lucidi 5~20, Rhizoma Polygonati 5~15, Herba Gueldenstaedtiae or Herba Violae 5~20, Radix Trichosanthis 5~20.
The preparation method of described Chinese medicine preparation: according to above-mentioned parts by weight, (1) Placenta Hominis is ground into 80 ~ 100 purpose powder, and is standby; (2) Radix Astragali, Radix Rehmanniae Preparata, Radix Codonopsis, the Rhizoma Atractylodis Macrocephalae, Radix Angelicae Sinensis, Radix Glycyrrhizae, Fructus Ligustri Lucidi, Rhizoma Polygonati, Herba Gueldenstaedtiae or Herba Violae, Radix Trichosanthis ten flavor medical materials decoct with water 1~3 time, each 1~3 hour, make thick paste or dried cream; (3) in thick paste or dried cream, add the Placenta Hominis powder, make suitable dosage form.
Further, every gram contains the crude drug amount and is equivalent to Placenta Hominis 0.15~0.18g, Radix Rehmanniae Preparata 0.45~0.55g, Fructus Ligustri Lucidi 0.25~0.35g, Radix Trichosanthis 0.25~0.35g in the described Chinese medicine preparation.
Optimized technical scheme is: described discrimination method comprise following discrimination method any or any two or wantonly three kinds or all:
(1) discrimination method of Placenta Hominis in the described Chinese medicine preparation is to be the thin layer chromatography discrimination method of contrast with the Placenta Hominis control medicinal material, may further comprise the steps:
1. the preparation of need testing solution: get described Chinese medicine preparation 3 ~ 10g, add chloroform or dichloromethane 20~30mL, supersound process 5~45 minutes, filter, concentrated filtrate is made per 1 mL and is contained the solution that the crude drug amount is equivalent to Placenta Hominis 0.2~5g, as need testing solution;
2. the preparation of control medicinal material solution: get Placenta Hominis control medicinal material 0.5 ~ 2g, add chloroform or dichloromethane 20~30mL, supersound process 5~45 minutes, filter, concentrated filtrate is made per 1 mL and is contained the solution that the crude drug amount is equivalent to Placenta Hominis 0.2~5g, in contrast medical material solution;
3. algoscopy: according to the test of an appendix VI of Chinese Pharmacopoeia version in 2010 B thin layer chromatography, drawing each 5~15 μ L of control medicinal material solution and need testing solution, put respectively on same silica gel g thin-layer plate, is developing solvent with normal hexane-ethyl acetate 9: 1 ~ 9, launch, take out, dry, spray perchloric acid solution with 5 ~ 50%, it is clear to be heated to speckle colour developing at 60 ~ 120 ℃, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the principal spot of same color;
(2) discrimination method of Radix Rehmanniae Preparata in the described Chinese medicine preparation is to be the thin layer chromatography discrimination method of contrast with the Radix Rehmanniae Preparata control medicinal material, may further comprise the steps:
1. the preparation of need testing solution: get described Chinese medicine preparation 6~15g, add water 100~200mL, warmly make fully molten loosing, be heated to and boil, put cold, centrifugal, get supernatant, be concentrated into 30~100mL, extract 1~3 time with the ethyl acetate jolting, each 20~30mL, combined ethyl acetate liquid, evaporate to dryness, residue add methanol makes dissolving, make per 1 mL and contain the solution that the crude drug amount is equivalent to Radix Rehmanniae Preparata 2~20g, as need testing solution;
2. the preparation of control medicinal material solution: get Radix Rehmanniae Preparata control medicinal material 2~10g, decoct with water 1 ~ 2 time, add water 50~150mL at every turn, the each decoction 0.5~2 hour filters merging filtrate with absorbent cotton, be concentrated into 30~100mL, extract 1~3 time each 20~30mL, combined ethyl acetate liquid with the ethyl acetate jolting, evaporate to dryness, residue adds methanol makes dissolving, makes per 1 mL and contains the solution that the crude drug amount is equivalent to Radix Rehmanniae Preparata 2~20g, in contrast medical material solution;
3. algoscopy: according to the test of an appendix VI of Chinese Pharmacopoeia version in 2010 B thin layer chromatography, draw each 5~15 μ L of control medicinal material solution and need testing solution, put respectively on same silica gel g thin-layer plate, with o-Dimethylbenzene-ethyl acetate 1:1 is developing solvent, wherein o-Dimethylbenzene is replaceable is benzene, or toluene, or meta-xylene, or xylol, or o-Dimethylbenzene, meta-xylene, three kinds of mixture of pressing arbitrary proportion of xylol launch, and take out, dry, spray is with 0.1% 2,4 dinitrophenyl hydrazine alcoholic solution, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the principal spot of same color;
(3) discrimination method of Fructus Ligustri Lucidi in the described Chinese medicine preparation is to be the thin layer chromatography discrimination method of contrast with Fructus Ligustri Lucidi control medicinal material and oleanolic acid reference substance, may further comprise the steps:
1. the preparation of need testing solution: get described Chinese medicine preparation 1.5~12g, add chloroform or dichloromethane 20~100mL, supersound process 5~45 minutes, filter, filtrate evaporate to dryness, residue add methanol or add dehydrated alcohol-chloroform by the blended solution of 3:2, make dissolving at 10~50 ℃, make per 1 mL and contain the solution that the crude drug amount is equivalent to Fructus Ligustri Lucidi 0.2~5g, as need testing solution;
2. the preparation of control medicinal material solution: get Fructus Ligustri Lucidi control medicinal material 0.5g, add chloroform or dichloromethane 20mL, supersound process 5 ~ 45 minutes, filter, filtrate evaporate to dryness, residue add methanol or add dehydrated alcohol-chloroform by the blended solution of 3:2, make dissolving at 10~50 ℃, make 1 mL and contain the solution that the crude drug amount is equivalent to Fructus Ligustri Lucidi 0.2~5g, in contrast medical material solution;
3. the preparation of reference substance solution: even up pier fruit acid reference substance, at 10~50 ℃, add methanol or 95% ethanol is mixed with the solution that every 1mL contains 1mg, in contrast product solution;
4. algoscopy: according to the test of an appendix VI of Chinese Pharmacopoeia version in 2010 B thin layer chromatography, at 10~50 ℃, draw need testing solution 5~15 μ L, each 2~5 μ L of control medicinal material solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with cyclohexane extraction-acetone-ethyl acetate-glacial acetic acid 7~15: 2~7: 1~5:0~0.5, be developing solvent perhaps with chloroform-methanol-formic acid 20~40:1:0.3~1.1, launch, take out, dry, spray is with 2 ~ 20% ethanol solution of sulfuric acid, perhaps spray with 1 ~ 10% phosphomolybdic acid ethanol solution, it is clear to be heated to speckle colour developing at 60~120 ℃, in the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the principal spot of same color;
(4) discrimination method of Radix Trichosanthis in the described Chinese medicine preparation is to be the thin layer chromatography discrimination method of contrast with Radix Trichosanthis control medicinal material and Citruline reference substance, may further comprise the steps:
1. the preparation of need testing solution: get described Chinese medicine preparation 6~12g, add 30%~100% ethanol, 20~50mL, supersound process 5~45 minutes, filter, the filtrate evaporate to dryness adds 50% ethanol and makes dissolving, filter, make per 1 mL and contain the solution that the crude drug amount is equivalent to Radix Trichosanthis 0.01~1g, as need testing solution;
2. the preparation of control medicinal material solution: get Radix Trichosanthis control medicinal material 2g, add 50% ethanol, supersound process 5 ~ 45 minutes filters, and makes per 1 mL and contains the solution that the crude drug amount is equivalent to Radix Trichosanthis 0.01~1g, in contrast medical material solution;
3. the preparation of reference substance solution: get the Citruline reference substance, add 50% ethanol and be mixed with the solution that every 1mL contains 1mg, in contrast product solution;
4. algoscopy: according to the test of an appendix VI of Chinese Pharmacopoeia version in 2010 B thin layer chromatography, draw need testing solution 5~15 μ L, each 1~5 μ L of control medicinal material solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with n-butyl alcohol-dehydrated alcohol-glacial acetic acid-water 7~15: 0~2:2 ~ 6:1~5, n-butyl alcohol-formic acid-water 15~25: 3:2, n-butyl alcohol-acetone-water 2~5: any among the 2:1 is developing solvent, launch, take out, dry, spray is with 1 ~ 10% ethanol solution of ninhydrin, it is clear to be heated to speckle colour developing at 60 ~ 120 ℃, in the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the principal spot of same color.
Preferred technical scheme is: described discrimination method comprise following discrimination method any or any two or wantonly three kinds or all:
(1) discrimination method of Placenta Hominis in the described Chinese medicine preparation may further comprise the steps:
Get described Chinese medicine preparation 6g, add chloroform 20mL, supersound process 30 minutes filters, and filtrate is concentrated into 1mL, as need testing solution; Other gets Placenta Hominis control medicinal material 1g, shines medical material solution in pairs with legal system; Test according to an appendix VI of Chinese Pharmacopoeia version in 2010 B thin layer chromatography, draw each 10 μ L of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with normal hexane-ethyl acetate is developing solvent at 9: 5, launch, take out, dry, spray is with 20% perchloric acid solution, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the principal spot of same color;
(2) discrimination method of Radix Rehmanniae Preparata in the described Chinese medicine preparation may further comprise the steps:
Get described Chinese medicine preparation 10g, add water 150mL, warmly make fully molten loosing, be heated to and boil, put cold, centrifugal, get supernatant, be concentrated into 100mL, extract 2 times each 30mL with the ethyl acetate jolting, combined ethyl acetate liquid, evaporate to dryness, residue add methanol 0.5mL makes dissolving, as need testing solution; Other gets Radix Rehmanniae Preparata control medicinal material 5g, decocts with water 2 times, adds water 80mL at every turn, 2 hours for the first time, 1 hour for the second time, filter with absorbent cotton, merging filtrate, be concentrated into 100mL, extract 2 times, each 30mL with the ethyl acetate jolting, combined ethyl acetate liquid, evaporate to dryness, residue add methanol 0.5mL makes dissolving, in contrast medical material solution; Test according to an appendix VI of Chinese Pharmacopoeia version in 2010 B thin layer chromatography, 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 o-Dimethylbenzene-ethyl acetate is developing solvent at 1: 1, launch, take out, dry, spray is with 0.1% 2,4 dinitrophenyl hydrazine alcoholic solution; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the principal spot of same color;
(3) discrimination method of Fructus Ligustri Lucidi in the described Chinese medicine preparation may further comprise the steps:
Get described Chinese medicine preparation 6g, add chloroform 20mL, supersound process 30 minutes filters, and filtrate evaporate to dryness, residue add methanol 1mL, make dissolving at 35~40 ℃, as need testing solution; Other gets Fructus Ligustri Lucidi control medicinal material 0.5g, shines medical material solution in pairs with legal system; Even up pier fruit acid reference substance again, add methanol and be mixed with the solution that every 1mL contains 1mg, in contrast product solution at 35~40 ℃; Test according to an appendix VI of Chinese Pharmacopoeia version in 2010 B thin layer chromatography, at 35~40 ℃, draw each 5 μ L of need testing solution 10 ~ 15 μ L, control medicinal material solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with cyclohexane extraction-acetone-ethyl acetate-glacial acetic acid 12.5: 5: 3:0.1 was developing solvent, launches, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to speckle colour developing at 110 ℃; In the test sample chromatograph, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, show the principal spot of same color;
(4) discrimination method of Radix Trichosanthis in the described Chinese medicine preparation may further comprise the steps:
Get described Chinese medicine preparation 10g, add 50% ethanol 30mL, supersound process 30 minutes filters, and the filtrate evaporate to dryness adds 50% ethanol 10mL and makes dissolving, filters, and gets filtrate as need testing solution; Other gets Radix Trichosanthis control medicinal material 2g, adds 50% ethanol 20mL, and supersound process 30 minutes filters, and gets filtrate medical material solution in contrast; Get the Citruline reference substance again, add 50% ethanol and make the solution that every 1mL contains 1mg, in contrast product solution; Test according to an appendix VI of Chinese Pharmacopoeia version in 2010 B thin layer chromatography, draw each 2 μ L of need testing solution 10 μ L, control medicinal material solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with n-butyl alcohol-dehydrated alcohol-glacial acetic acid-water 8: 2:4:3 is developing solvent, launch, take out, dry, spray is with 2% ethanol solution of ninhydrin, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, show the principal spot of same color.
In the above-mentioned technical scheme that relates to, in the discrimination method about Fructus Ligustri Lucidi, can take to serve as contrast separately or be the thin layer chromatography discrimination method of contrast with the oleanolic acid reference substance separately with the Fructus Ligustri Lucidi control medicinal material; In the discrimination method about Radix Trichosanthis, can take to serve as contrast separately or be the thin layer chromatography discrimination method of contrast with the Citruline reference substance separately with the Radix Trichosanthis control medicinal material.
The discrimination method of Chinese medicine preparation provided by the invention, control medicinal material be main tlc identification method be one practicable, easy, fast, controlled, economic, science, modern discrimination method.The present invention is by comparing several different methods such as ultraviolet visible spectrophotometry, paper chromatography, thin layer chromatography, high performance liquid chromatography, having filtered out with control medicinal material, chemical reference substance is the silica gel G thin layer chromatography of contrast, differentiates the method for Placenta Hominis in the Chinese medicine preparation of the present invention, Radix Rehmanniae Preparata, Fructus Ligustri Lucidi, Radix Trichosanthis.Placenta Hominis, Radix Rehmanniae Preparata, Fructus Ligustri Lucidi, Radix Trichosanthis are the main flavour of a drug of this Chinese medicine preparation, and discrimination method specificity provided by the invention is strong, and repeatability, ruggedness are good, can control the quality of Chinese medicine preparation of the present invention preferably.
The specific embodiment
Embodiment 1
Prescription: Placenta Hominis 50g, Radix Astragali 300g, Radix Rehmanniae Preparata 150g, Radix Codonopsis 100g, Rhizoma Atractylodis Macrocephalae 100g, Radix Angelicae Sinensis 75g, Radix Glycyrrhizae 75g, Fructus Ligustri Lucidi 100g, Rhizoma Polygonati 75g, Herba Gueldenstaedtiae 100g, Radix Trichosanthis 100g.
Method for making: above ten simply, and Placenta Hominis is ground into 80 ~ 100 purpose powder, and ten flavors such as all the other Radixs Astragali decoct with water twice, 2 hours for the first time, 1 hour for the second time, collecting decoction, filter, it is 1.32~1.35(50 ℃ that filtrate is concentrated into relative density) thick paste, add the Placenta Hominis powder, mixing, granulate, incapsulate, every dress 0.3g, make 1000, promptly.
Differentiate: the discriminating of this capsule comprise following discrimination method any or any two or wantonly three kinds or whole, but be not limited only to following discrimination method:
(1) get this product content 6g, add chloroform 20mL, supersound process 30 minutes filters, and filtrate is concentrated into 1mL, as need testing solution.Other gets Placenta Hominis control medicinal material 1g, shines medical material solution in pairs with legal system.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), draw each 10 μ L of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with normal hexane-ethyl acetate (9: 5) is developing solvent, launch, take out, dry, spray is with 20% perchloric acid solution, and it is clear to be heated to the speckle colour developing at 105 ℃.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the principal spot of same color.
(2) get this product content 10g, add water 150mL, warmly make fully molten loosing, be heated to and boil, put cold, centrifugal, get supernatant, be concentrated into 100mL, extract 2 times each 30mL with the ethyl acetate jolting, combined ethyl acetate liquid, evaporate to dryness, residue add methanol 0.5mL makes dissolving, as need testing solution.Other gets Radix Rehmanniae Preparata control medicinal material 5g, decocts with water 2 times, adds water 80mL at every turn, 2 hours for the first time, 1 hour for the second time, filter with absorbent cotton, merging filtrate, be concentrated into 100mL, extract 2 times, each 30mL with the ethyl acetate jolting, combined ethyl acetate liquid, evaporate to dryness, residue add methanol 0.5mL makes dissolving, in contrast medical material solution.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), 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 o-Dimethylbenzene-ethyl acetate (1: 1) is developing solvent, launch, take out, dry, spray is with 0.1% 2,4 dinitrophenyl hydrazine alcoholic solution.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the principal spot of same color.
Wherein: 0.1% 2,4 dinitrophenyl hydrazine alcoholic solution compound method: get 2,4 dinitrophenyl hydrazine 1g, add dehydrated alcohol 1000 mL and make dissolving, slowly add concentration again and be 36%~38% hydrochloric acid 10mL, shake up, promptly.
(3) get need testing solution under above-mentioned (1), evaporate to dryness, residue add methanol 1mL, make dissolving at 35~40 ℃, as need testing solution.Other gets Fructus Ligustri Lucidi control medicinal material 0.5g, adds chloroform 20mL, and supersound process 30 minutes filters, and filtrate evaporate to dryness, residue add methanol 1mL, make dissolving at 35~40 ℃, in contrast medical material solution.Even up pier fruit acid reference substance again, add methanol, be mixed with the solution that every 1mL contains 1mg at 35~40 ℃, in contrast product solution.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), at 35~40 ℃, draw each 5 μ L of need testing solution 10 μ L, control medicinal material solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with cyclohexane extraction-acetone-ethyl acetate-glacial acetic acid (12.5: 5: 3:0.1) be developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to speckle colour developing at 110 ℃.In the test sample chromatograph, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, show the principal spot of same color.
(4) get this product content 10g, add 50% ethanol 30mL, supersound process 30 minutes filters, and the filtrate evaporate to dryness adds 50% ethanol 10mL and makes dissolving, filters, and gets filtrate as need testing solution.Other gets Radix Trichosanthis control medicinal material 2g, adds 50% ethanol 20mL, and supersound process 30 minutes filters, and gets filtrate medical material solution in contrast.Get the Citruline reference substance again, add 50% ethanol, be mixed with the solution that every 1mL contains 1mg, in contrast product solution.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), draw each 2 μ L of need testing solution 10 μ L, control medicinal material solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with n-butyl alcohol-dehydrated alcohol-glacial acetic acid-water (8: 2:4:3) be developing solvent, launch, take out, dry, spray is with 2% ethanol solution of ninhydrin, and it is clear to be heated to the speckle colour developing at 105 ℃.In the test sample chromatograph, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, show the principal spot of same color.
Embodiment 2
Prescription: Placenta Hominis 50g, Radix Astragali 300g, Radix Rehmanniae Preparata 150g, Radix Codonopsis 100g, Rhizoma Atractylodis Macrocephalae 100g, Radix Angelicae Sinensis 80g, Radix Glycyrrhizae 80g, Fructus Ligustri Lucidi 80g, Rhizoma Polygonati 100g, Herba Violae 100g, Radix Trichosanthis 100g.
Method for making: above ten simply, and Placenta Hominis is ground into 80 ~ 100 purpose powder, and ten flavors such as all the other Radixs Astragali decoct with water twice, 2 hours for the first time, 1 hour for the second time, collecting decoction, filter, filtrate is condensed into thick paste, further is dried to dried cream, be ground into 80 ~ 100 purpose powder again, add the Placenta Hominis powder, mixing, granulate tabletting, every heavy 0.3g, the bag film-coat is made 1000, promptly.
Differentiate: the discriminating of this tablet comprise following discrimination method any or any two or wantonly three kinds or whole, but be not limited only to following discrimination method:
(1) get the powder 6g that this product grinds, add chloroform 25mL, supersound process 20 minutes filters, and residue is standby, and filtrate is concentrated into 0.5mL, as need testing solution.Other gets Placenta Hominis control medicinal material 1g, shines medical material solution in pairs with legal system.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), draw each 15 μ L of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with normal hexane-ethyl acetate (9: 4) is developing solvent, launch, take out, dry, spray is with 20% perchloric acid solution, and it is clear to be heated to the speckle colour developing at 105 ℃.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the principal spot of same color.
(2) get the powder 6g that this product grinds, add water 100mL, warmly make fully molten loosing, be heated to and boil, put cold, centrifugal, get supernatant, extract 2 times each 30mL with the ethyl acetate jolting, combined ethyl acetate liquid, evaporate to dryness, residue add methanol 0.5mL makes dissolving, as need testing solution.Other gets Radix Rehmanniae Preparata control medicinal material 4g, adds water 60mL, decocts 30 minutes, filters with absorbent cotton, and filtrate is extracted 2 times with the ethyl acetate jolting, each 20mL, and combined ethyl acetate liquid, evaporate to dryness, residue add methanol 0.5mL makes dissolving, in contrast medical material solution.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), draw need testing solution 15 μ L, control medicinal material solution 5 μ L, put respectively on same silica gel g thin-layer plate, with dimethylbenzene-ethyl acetate (1: 1) is developing solvent, launch, take out, dry, spray is with 0.1% 2,4 dinitrophenyl hydrazine alcoholic solution.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the principal spot of same color.
Wherein: 1. 0.1% 2,4 dinitrophenyl hydrazine alcoholic solution compound method: get 2,4 dinitrophenyl hydrazine 1g, add dehydrated alcohol 1000 mL and make dissolving, slowly add concentration again and be 36%~38% hydrochloric acid 10mL, shake up, promptly; 2. dimethylbenzene is o-Dimethylbenzene, meta-xylene, three kinds of mixture of isomers of xylol.
(3) get the powder 1.5g that this product grinds, add chloroform 20mL, supersound process 30 minutes filters, and filtrate evaporate to dryness, residue add methanol 1mL, make dissolving at 30~35 ℃, as need testing solution.Other gets Fructus Ligustri Lucidi control medicinal material 0.5g, shines medical material solution in pairs with legal system.Even up pier fruit acid reference substance again, add methanol, be mixed with the solution that every 1mL contains 1mg at 30~35 ℃, in contrast product solution.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), at 30~35 ℃, draw each 4 μ L of need testing solution 15 μ L, control medicinal material solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with cyclohexane extraction-acetone-ethyl acetate-glacial acetic acid (12.5: 5: 3:0.1) be developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to speckle colour developing at 110 ℃.In the test sample chromatograph, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, show the principal spot of same color.
(4) get above-mentioned (1) item standby residue down, add 60% ethanol 30mL, supersound process 30 minutes filters, and the filtrate evaporate to dryness adds 50% ethanol 10mL to make dissolving, filters, and gets filtrate as need testing solution.Other gets Radix Trichosanthis control medicinal material 2g, adds 50% ethanol 20mL, and supersound process 30 minutes filters, and gets filtrate medical material solution in contrast.Get the Citruline reference substance again, add 50% ethanol, be mixed with the solution that every 1mL contains 1mg, in contrast product solution.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), draw each 2 μ L of need testing solution 15 μ L, control medicinal material solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with n-butyl alcohol-dehydrated alcohol-glacial acetic acid-water (8: 2:5:3) be developing solvent, launch, take out, dry, spray is with 2% ethanol solution of ninhydrin, and it is clear to be heated to the speckle colour developing at 105 ℃.In the test sample chromatograph, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, show the principal spot of same color.
Embodiment 3
Prescription: Placenta Hominis 50g, Radix Astragali 300g, Radix Rehmanniae Preparata 150g, Radix Codonopsis 100g, Rhizoma Atractylodis Macrocephalae 100g, Radix Angelicae Sinensis 75g, Radix Glycyrrhizae 75g, Fructus Ligustri Lucidi 100g, Rhizoma Polygonati 75g, Herba Violae 100g, Radix Trichosanthis 100g.
Method for making: above ten simply, and Placenta Hominis is ground into 80 ~ 100 purpose powder, and ten flavors such as all the other Radixs Astragali decoct with water twice, 2 hours for the first time, 1 hour for the second time, collecting decoction, filter, it is 1.32~1.35(50 ℃ that filtrate is concentrated into relative density) thick paste, add the Placenta Hominis powder, mixing, pill, drying, polishing, make 2000 concentrated pills, every heavy 0.15g, promptly.
Differentiate: the discriminating of this concentrated pill comprise following discrimination method any or any two or wantonly three kinds or whole, but be not limited to following discrimination method:
(1) get the powder 6g that this product grinds, the 30mL that adds methylene chloride, supersound process 25 minutes filters, and filtrate is concentrated into 1mL, as need testing solution.Other gets Placenta Hominis control medicinal material 1g, shines medical material solution in pairs with legal system.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), draw each 5 μ L of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with normal hexane-ethyl acetate (9: 3) is developing solvent, launch, take out, dry, spray is with 20% perchloric acid solution, and it is clear to be heated to the speckle colour developing at 105 ℃.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the principal spot of same color.
(2) get the powder 6g that this product grinds, add water 100mL, warmly make fully molten loosing, be heated to and boil, put cold, centrifugal, get supernatant, be concentrated into 40mL, extract 2 times each 20mL with the ethyl acetate jolting, combined ethyl acetate liquid, evaporate to dryness, residue add methanol 1mL makes dissolving, as need testing solution.Other gets Radix Rehmanniae Preparata control medicinal material 4g, adds water 60mL, decocts 30 minutes, filters with absorbent cotton, and filtrate is extracted 2 times with the ethyl acetate jolting, each 20 mL, and combined ethyl acetate liquid, evaporate to dryness, residue add methanol 1mL makes dissolving, in contrast medical material solution.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), 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 toluene-ethyl acetate (1: 1) is developing solvent, launch, take out, dry, spray is with 0.1% 2,4 dinitrophenyl hydrazine alcoholic solution.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the principal spot of same color.
Wherein: 0.1% 2,4 dinitrophenyl hydrazine alcoholic solution compound method: get 2,4 dinitrophenyl hydrazine 1g, add dehydrated alcohol 1000 mL and make dissolving, slowly add concentration again and be 36%~38% hydrochloric acid 10mL, shake up, promptly.
(3) get the powder 10g that this product grinds, add chloroform 30mL, supersound process 20 minutes filters, and filtrate evaporate to dryness, residue add methanol 1mL, make dissolving at 25~30 ℃, as need testing solution.Other gets Fructus Ligustri Lucidi control medicinal material 0.5g, shines medical material solution in pairs with legal system.Even up pier fruit acid reference substance again, add methanol, be mixed with the solution that every 1mL contains 1mg at 25~30 ℃, in contrast product solution.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), at 25~30 ℃, draw need testing solution 15 μ L, control medicinal material solution 2 μ L, reference substance solution 5 μ L, putting respectively on same silica gel g thin-layer plate, is developing solvent with chloroform-methanol-formic acid (25:1:1), launches, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to speckle colour developing at 105 ℃.In the test sample chromatograph, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, show the principal spot of same color.
(4) get the powder 12g that this product grinds, add 70% ethanol 50mL, supersound process 20 minutes filters, and the filtrate evaporate to dryness adds 50% ethanol 5mL and makes dissolving, filters, and gets filtrate as need testing solution.Other gets Radix Trichosanthis control medicinal material 2g, adds 50% ethanol 20mL, and supersound process 30 minutes filters, and gets filtrate medical material solution in contrast.Get the Citruline reference substance again, add 50% ethanol, be mixed with the solution that every 1mL contains 1mg, in contrast product solution.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), draw each 2 μ L of need testing solution 5 μ L, control medicinal material solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with n-butyl alcohol-glacial acetic acid-water (13: 3:4) be developing solvent, launch, take out, dry, spray is with 2% ethanol solution of ninhydrin, and it is clear to be heated to the speckle colour developing at 105 ℃.In the test sample chromatograph, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, show the principal spot of same color.
Embodiment 4
Prescription: Placenta Hominis 50g, Radix Astragali 300g, Radix Rehmanniae Preparata 150g, Radix Codonopsis 100g, Rhizoma Atractylodis Macrocephalae 100g, Radix Angelicae Sinensis 75g, Radix Glycyrrhizae 75g, Fructus Ligustri Lucidi 100g, Rhizoma Polygonati 75g, Herba Gueldenstaedtiae 100g, Radix Trichosanthis 100g.
Method for making: above ten simply, and Placenta Hominis is ground into 80 ~ 100 purpose powder, and ten flavors such as all the other Radixs Astragali decoct with water twice, 2 hours for the first time, 1 hour for the second time, collecting decoction filtered, filtrate is concentrated into half amount, adds 95% ethanol of same capability, places 12 hours for 2~10 ℃, filter, being concentrated into relative density behind the filtrate recycling ethanol is 1.29~1.31(50 ℃) thick paste, add the Placenta Hominis powder, mixing is granulated, and incapsulates, every dress 0.3g makes 1000, promptly.
Differentiate: the discriminating of this capsule comprise following discrimination method any or all, but be not limited only to following discrimination method:
(1) get this product content 12g, the 100mL that adds methylene chloride, supersound process 10 minutes filters, and filtrate is concentrated into 1mL, and evaporate to dryness, residue add dehydrated alcohol-chloroform by the blended solution 1mL of 3:2, make dissolving at 20~25 ℃, as need testing solution.Other gets Fructus Ligustri Lucidi control medicinal material 1g, shines medical material solution in pairs with legal system.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), at 20~25 ℃, 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 cyclohexane extraction-acetone-ethyl acetate (10:4: 2.5) be developing solvent, launch, take out, dry, spray is with 5% phosphomolybdic acid ethanol solution, and is clear to the speckle colour developing 105 ℃ of heat.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the principal spot of same color.
(2) get this product content 10g, add 30% ethanol 20mL, supersound process 5 minutes filters, and the filtrate evaporate to dryness adds 50% ethanol 10mL and makes dissolving, filters, and gets filtrate as need testing solution.Other gets the Citruline reference substance, adds 50% ethanol, is mixed with the solution that every 1mL contains 1mg, in contrast product solution.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), draw need testing solution 10 μ L, reference substance solution 2 μ L, put respectively on same silica gel g thin-layer plate, with n-butyl alcohol-acetone-water (4: 2:1) be developing solvent, launch, take out, dry, spray is with 2% ethanol solution of ninhydrin, and it is clear to be heated to the speckle colour developing at 105 ℃.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
Embodiment 5
Prescription and three batches of capsules of method for making preparation by embodiment 1 capsule carry out the methodology checking:
(1) the methodology checking of Placenta Hominis discriminating:
The preparation of control medicinal material solution: get Placenta Hominis control medicinal material 1g, add chloroform 20mL, supersound process 30 minutes filters, and filtrate is concentrated into 1mL, in contrast medical material solution.
The preparation of need testing solution: get capsule 's content 6g, add chloroform 20mL, supersound process 30 minutes filters, and filtrate is concentrated into 1mL, as need testing solution.Method prepares the need testing solution of three batches of capsules according to this.
The preparation of negative control solution: in the negative control sample of capsule prescription ratio and the scarce Placenta Hominis of prepared, get negative control sample 6g, add chloroform 20mL, supersound process 30 minutes filters, and filtrate is concentrated into 1mL, as negative control solution.
Algoscopy: according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B) test, draw control medicinal material solution, negative control solution and three crowdes of each 10 μ L of the capsular need testing solution of different batches, put respectively on same silica gel g thin-layer plate, with normal hexane-ethyl acetate (9: 5) is developing solvent, launch, take out, dry, spray is with 20% perchloric acid solution, and it is clear to be heated to the speckle colour developing at 105 ℃.
Result of the test: in three batches of test sample chromatographs, with the corresponding position of control medicinal material chromatograph on, show the principal spot of same color; And in the negative control chromatograph, with the corresponding position of control medicinal material chromatograph on, display dot not.
Conclusion: this method specificity is strong, good reproducibility, can be used for the discriminating of Placenta Hominis in this capsule.
(2) the methodology checking of Radix Rehmanniae Preparata discriminating:
The preparation of control medicinal material solution: get Radix Rehmanniae Preparata control medicinal material 5g, decoct with water 2 times, add water 80mL at every turn, 2 hours for the first time, 1 hour for the second time, filter with absorbent cotton, merging filtrate is concentrated into 100mL, extract 2 times with the ethyl acetate jolting, each 30mL, combined ethyl acetate liquid, evaporate to dryness, residue adds methanol 0.5mL makes dissolving, in contrast medical material solution.
The preparation of need testing solution: get capsule 's content 10g, add water 150mL, warmly make fully molten loosing, be heated to and boil, put cold, centrifugal (2000 change) 5 minutes, get supernatant, be concentrated into 100mL, extract 2 times, each 30mL with the ethyl acetate jolting, combined ethyl acetate liquid, evaporate to dryness, residue add methanol 0.5mL makes dissolving, as need testing solution.Method prepares the need testing solution of three batches of capsules according to this.
The preparation of negative control solution: in the negative control sample of capsule prescription ratio and the scarce Radix Rehmanniae Preparata of prepared, get negative control sample 10g, add water 150mL, warmly make fully molten loosing, be heated to and boil, put cold, centrifugal (2000 change) 5 minutes gets supernatant, is concentrated into 100mL, extract 2 times with the ethyl acetate jolting, each 30mL, combined ethyl acetate liquid, evaporate to dryness, residue adds methanol 0.5mL makes dissolving, as negative control solution.
Algoscopy: according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B) test, draw control medicinal material solution 5 μ L, negative control solution and three crowdes of each 10 μ L of the capsular need testing solution of different batches, put respectively on same silica gel g thin-layer plate, with o-Dimethylbenzene-ethyl acetate (1: 1) is developing solvent, launch, take out, dry, spray is with 0.1% 2,4 dinitrophenyl hydrazine alcoholic solution.Wherein: 0.1% 2,4 dinitrophenyl hydrazine alcoholic solution compound method: get 2,4 dinitrophenyl hydrazine 1g, add dehydrated alcohol 1000 mL and make dissolving, slowly add concentration again and be 36%~38% hydrochloric acid 10mL, shake up, promptly.
Result of the test: in three batches of test sample chromatographs, with the corresponding position of control medicinal material chromatograph on, show the principal spot of same color; And in the negative control chromatograph, with the corresponding position of control medicinal material chromatograph on, display dot not.
Conclusion: this method specificity is strong, good reproducibility, can be used for the discriminating of Radix Rehmanniae Preparata in this capsule.
(3) the methodology checking of Fructus Ligustri Lucidi discriminating:
The preparation of control medicinal material solution: get Fructus Ligustri Lucidi control medicinal material 0.5g, add chloroform 20mL, supersound process 30 minutes filters, and filtrate evaporate to dryness, residue add methanol 1mL, and 35~40 ℃ make dissolving, in contrast medical material solution.
The preparation of reference substance solution: even up pier fruit acid reference substance, add methanol, be mixed with the solution that every 1mL contains 1mg at 35~40 ℃, in contrast product solution.
The preparation of need testing solution: get capsule 's content 6g, add chloroform 20mL, supersound process 30 minutes filters, and filtrate evaporate to dryness, residue add methanol 1mL, and 35~40 ℃ make dissolving, as need testing solution.Method prepares the need testing solution of three batches of capsules according to this.
The preparation of negative control solution: in the negative control sample of capsule prescription ratio and the scarce Fructus Ligustri Lucidi of prepared, get negative control sample 6g, add chloroform 20mL, supersound process 30 minutes filters the filtrate evaporate to dryness, residue adds methanol 1mL, and 35~40 ℃ make dissolving, as negative control solution.
Algoscopy: according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B) test, at 35~40 ℃, draw each 5 μ L of control medicinal material solution and reference substance solution, negative control solution and three crowdes of each 10 μ L of the capsular need testing solution of different batches, put respectively on same silica gel g thin-layer plate, with cyclohexane extraction-acetone-ethyl acetate-glacial acetic acid (12.5: 5: 3:0.1) be developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 110 ℃.
Result of the test: in three batches of test sample chromatographs, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, show the principal spot of same color; And in the negative control chromatograph, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, display dot not.
Conclusion: this method specificity is strong, good reproducibility, can be used for the discriminating of Fructus Ligustri Lucidi in this capsule.
(4) the methodology checking of Radix Trichosanthis discriminating:
The preparation of control medicinal material solution: get Radix Trichosanthis control medicinal material 2g, add 50% ethanol 20mL, supersound process 30 minutes filters, and gets filtrate medical material solution in contrast.
The preparation of reference substance solution: get the Citruline reference substance, add 50% ethanol, be mixed with the solution that every 1mL contains 1mg, in contrast product solution.
The preparation of need testing solution: get capsule 's content 10g, add 50% alcoholic solution 20mL, supersound process 30 minutes filters, and the filtrate evaporate to dryness adds 50% ethanol 10mL and makes dissolving, filters, and gets filtrate as need testing solution.Method prepares the need testing solution of three batches of capsules according to this.
The preparation of negative control solution: in the negative control sample of capsule prescription ratio and the scarce Radix Trichosanthis of prepared, get negative control sample 10g, add 50% ethanol 20mL, supersound process 30 minutes filters the filtrate evaporate to dryness, add 50% ethanol 10mL and make dissolving, filter, get filtrate as need testing solution.
Algoscopy: according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B) test, draw each 2 μ L of control medicinal material solution and reference substance solution, negative control solution and three crowdes of each 10 μ L of the capsular need testing solution of different batches, put respectively on same silica gel g thin-layer plate, with n-butyl alcohol-dehydrated alcohol-glacial acetic acid-water (8: 2:4:3) be developing solvent, launch, take out, dry, spray is with 2% ethanol solution of ninhydrin, and it is clear to be heated to the speckle colour developing at 105 ℃.
Result of the test: in three batches of test sample chromatographs, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, show the principal spot of same color; And in the negative control chromatograph, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, display dot not.
Conclusion: this method specificity is strong, good reproducibility, can be used for the discriminating of Radix Trichosanthis in this capsule.
The discrimination method of other situations of mentioning among the present invention has also carried out above-mentioned corresponding methodology checking, proves that the same specificity of discrimination method of other situations is strong, good reproducibility, can be used for corresponding discriminating.
Embodiment 6
Prescription and three batches of tablets of method for making preparation by embodiment 2 tablets carry out the methodology checking:
(1) the methodology checking of Placenta Hominis discriminating:
The preparation of control medicinal material solution: get Placenta Hominis control medicinal material 1g, add chloroform 25mL, supersound process 20 minutes filters, and filtrate is concentrated into 0.5mL, in contrast medical material solution.
The preparation of need testing solution: get the powder 6g that this product grinds, add chloroform 25mL, supersound process 20 minutes filters, and filtrate is concentrated into 0.5mL, as need testing solution.Method prepares the need testing solution of three batches of tablets according to this.
The preparation of negative control solution: the negative control sample that lacks Placenta Hominis in embodiment 2 tablet formulation ratios and prepared.Get negative control sample 6g, add chloroform 25mL, supersound process 20 minutes filters, and filtrate is concentrated into 0.5mL, as negative control solution.
Algoscopy: according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B) test, draw each 15 μ L of need testing solution of control medicinal material solution, negative control solution and three batches of different batches tablets, put respectively on same silica gel g thin-layer plate, with normal hexane-ethyl acetate (9: 4) is developing solvent, launch, take out, dry, spray is with 20% perchloric acid solution, and it is clear to be heated to the speckle colour developing at 105 ℃.
Result of the test: in three batches of test sample chromatographs, with the corresponding position of control medicinal material chromatograph on, show the principal spot of same color; And in the negative control chromatograph, with the corresponding position of control medicinal material chromatograph on, display dot not.
Conclusion: this method specificity is strong, good reproducibility, can be used for the discriminating of Placenta Hominis in this tablet.
(2) the methodology checking of Radix Rehmanniae Preparata discriminating:
The preparation of control medicinal material solution: get Radix Rehmanniae Preparata control medicinal material 4g, add water 60mL and decocted 30 minutes, filter with absorbent cotton, extract 2 times each 20mL, combined ethyl acetate liquid with the ethyl acetate jolting, evaporate to dryness, residue add methanol 0.5mL makes dissolving, in contrast medical material solution.
The preparation of need testing solution: get the powder 6g that this product grinds, add water 100mL, warmly make fully molten loosing, be heated to and boil, put cold, centrifugal, get supernatant, extract 2 times each 30mL with the ethyl acetate jolting, combined ethyl acetate liquid, evaporate to dryness, residue add methanol 0.5mL makes dissolving, as need testing solution.Method prepares the need testing solution of three batches of tablets according to this.
The preparation of negative control solution: the negative control sample that lacks Radix Rehmanniae Preparata in embodiment 2 tablet formulation ratios and prepared.Get the powder 6g that the negative control sample grinds, add water 100mL, warmly make fully molten loosing, be heated to and boil, put cold, centrifugal, get supernatant, extract 2 times each 30mL with the ethyl acetate jolting, combined ethyl acetate liquid, evaporate to dryness, residue add methanol 0.5mL makes dissolving, as negative control solution.
Algoscopy: according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B) test, draw need testing solution each 15 μ L, control medicinal material solution 5 μ L of negative control solution and three batches of different batches tablets, put respectively on same silica gel g thin-layer plate, with dimethylbenzene-ethyl acetate (1: 1) is developing solvent, launch, take out, dry, spray is with 0.1% 2,4 dinitrophenyl hydrazine alcoholic solution.
Wherein: 1. 0.1% 2,4 dinitrophenyl hydrazine alcoholic solution compound method: get 2,4 dinitrophenyl hydrazine 1g, add dehydrated alcohol 1000 mL and make dissolving, slowly add concentration again and be 36%~38% hydrochloric acid 10mL, shake up, promptly; 2. dimethylbenzene is o-Dimethylbenzene, meta-xylene, three kinds of mixture of isomers of xylol.
Result of the test: in three batches of test sample chromatographs, with the corresponding position of control medicinal material chromatograph on, show the principal spot of same color; And in the negative control chromatograph, with the corresponding position of control medicinal material chromatograph on, display dot not.
Conclusion: this method specificity is strong, good reproducibility, can be used for the discriminating of Radix Rehmanniae Preparata in this tablet.
(3) the methodology checking of Fructus Ligustri Lucidi discriminating:
The preparation of control medicinal material solution: get Fructus Ligustri Lucidi control medicinal material 0.5g, add chloroform 20mL, supersound process 30 minutes filters, and filtrate evaporate to dryness, residue add methanol 1mL, make dissolving at 30~35 ℃, in contrast medical material solution.
The preparation of reference substance solution: even up pier fruit acid reference substance, add methanol, be mixed with the solution that every 1mL contains 1mg at 30~35 ℃, in contrast product solution.
The preparation of need testing solution: get the powder 1.5g that this product grinds, add chloroform 20mL, supersound process 30 minutes filters, and filtrate evaporate to dryness, residue add methanol 1mL, make dissolving at 30~35 ℃, as need testing solution.Method prepares the need testing solution of three batches of tablets according to this.
The preparation of negative control solution: the negative control sample that lacks Fructus Ligustri Lucidi in embodiment 2 tablet formulation ratios and prepared.Get the powder 1.5g that the negative control sample grinds, add chloroform 20mL, supersound process 30 minutes filters, and filtrate evaporate to dryness, residue add methanol 1mL, make dissolving at 30~35 ℃, as negative control solution.
Algoscopy: according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B) test, at 30~35 ℃, draw each 15 μ L of need testing solution, control medicinal material solution and each 4 μ L of reference substance solution of negative control solution and three batches of different batches tablets, put respectively on same silica gel g thin-layer plate, with cyclohexane extraction-acetone-ethyl acetate-glacial acetic acid (12.5: 5: 3:0.1) be developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 110 ℃.
Result of the test: in three batches of test sample chromatographs, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, show the principal spot of same color; And in the negative control chromatograph, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, display dot not.
Conclusion: this method specificity is strong, good reproducibility, can be used for the discriminating of Fructus Ligustri Lucidi in this tablet.
(4) the methodology checking of Radix Trichosanthis discriminating:
The preparation of control medicinal material solution: get Radix Trichosanthis control medicinal material 2g, add 50% ethanol 20mL, supersound process 30 minutes filters, and gets subsequent filtrate medical material solution in contrast.
The preparation of reference substance solution: get the Citruline reference substance, add 50% ethanol, be mixed with the solution that every 1mL contains 1mg, in contrast product solution.
The preparation of need testing solution: get the powder 6g that this product grinds, add 60% ethanol 30mL, supersound process 30 minutes filters, and the filtrate evaporate to dryness adds 50% ethanol 10mL and makes dissolving, filters, and gets subsequent filtrate as need testing solution.Method prepares the need testing solution of three batches of tablets according to this.
The preparation of negative control solution: the negative control sample that lacks Radix Trichosanthis in embodiment 2 tablet formulation ratios and prepared.Get the powder 6g that the negative control sample grinds, add 60% ethanol 30mL, supersound process 30 minutes filters, and the filtrate evaporate to dryness adds 50% ethanol 10mL and makes dissolving, filters, and gets subsequent filtrate as need testing solution.
Algoscopy: according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B) test, draw each 15 μ L of need testing solution, control medicinal material solution and each 2 μ L of reference substance solution of negative control solution and three batches of different batches tablets, put respectively on same silica gel g thin-layer plate, with n-butyl alcohol-dehydrated alcohol-glacial acetic acid-water (8: 2:5:3) be developing solvent, launch, take out, dry, spray is with 2% ethanol solution of ninhydrin, and it is clear to be heated to the speckle colour developing at 105 ℃.
Result of the test: in three batches of test sample chromatographs, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, show the principal spot of same color; And in the negative control chromatograph, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, display dot not.
Conclusion: this method specificity is strong, good reproducibility, can be used for the discriminating of Radix Trichosanthis in this tablet.
Embodiment 7
Prescription and three batches of concentrated pills of method for making preparation by embodiment 3 concentrated pills carry out the methodology checking:
(1) the methodology checking of Placenta Hominis discriminating:
The preparation of control medicinal material solution: get Placenta Hominis control medicinal material 1g, the 30mL that adds methylene chloride, supersound process 25 minutes filters, and filtrate is concentrated into 1mL, in contrast medical material solution.
The preparation of need testing solution: get the powder 6g that this product grinds, the 30mL that adds methylene chloride, supersound process 25 minutes filters, and filtrate is concentrated into 1mL, as need testing solution.Method prepares the need testing solution of three batches of concentrated pills according to this.
The preparation of negative control solution: the negative control sample that lacks Placenta Hominis in embodiment 3 concentrated pills prescription ratio and prepared.Get the powder 6g that the negative control sample grinds, the 30mL that adds methylene chloride, supersound process 25 minutes filters, and filtrate is concentrated into 1mL, as negative control solution.
Algoscopy: according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B) test, draw each 5 μ L of need testing solution of control medicinal material solution, negative control solution and three batches of different batches concentrated pills, put respectively on same silica gel g thin-layer plate, with normal hexane-ethyl acetate (9: 3) is developing solvent, launch, take out, dry, spray is with 20% perchloric acid solution, and it is clear to be heated to the speckle colour developing at 105 ℃.
Result of the test: in three batches of test sample chromatographs, with the corresponding position of control medicinal material chromatograph on, show the principal spot of same color; And in the negative control chromatograph, with the corresponding position of control medicinal material chromatograph on, display dot not.
Conclusion: this method specificity is strong, good reproducibility, can be used for the discriminating of Placenta Hominis in this concentrated pill.
(2) the methodology checking of Radix Rehmanniae Preparata discriminating:
The preparation of control medicinal material solution: get Radix Rehmanniae Preparata control medicinal material 4g, add water 60mL, decocted 30 minutes, filter with absorbent cotton, filtrate is extracted 2 times with the ethyl acetate jolting, each 20 mL, combined ethyl acetate liquid, evaporate to dryness, residue add methanol 1mL makes dissolving, in contrast medical material solution.
The preparation of need testing solution: get the powder 6g that this product grinds, add water 100mL, warmly make fully molten loosing, be heated to and boil, put cold, centrifugal, get supernatant, be concentrated into 40mL, extract 2 times, each 20mL with the ethyl acetate jolting, combined ethyl acetate liquid, evaporate to dryness, residue add methanol 1mL makes dissolving, as need testing solution.Method prepares the need testing solution of three batches of concentrated pills according to this.
The preparation of negative control solution: the negative control sample that lacks Radix Rehmanniae Preparata in embodiment 3 concentrated pills prescription ratio and prepared.Get the powder 6g that the negative control sample grinds, add water 100mL, warmly make fully molten loosing, be heated to and boil, put cold, centrifugal, get supernatant, be concentrated into 40mL, extract 2 times, each 20mL with the ethyl acetate jolting, combined ethyl acetate liquid, evaporate to dryness, residue add methanol 1mL makes dissolving, as negative control solution.
Algoscopy: according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B) test, draw need testing solution each 10 μ L, control medicinal material solution 5 μ L of negative control solution and three batches of different batches concentrated pills, put respectively on same silica gel g thin-layer plate, with toluene-ethyl acetate (1: 1) is developing solvent, launch, take out, dry, spray is with 0.1% 2,4 dinitrophenyl hydrazine alcoholic solution.
Wherein: 0.1% 2,4 dinitrophenyl hydrazine alcoholic solution compound method: get 2,4 dinitrophenyl hydrazine 1g, add dehydrated alcohol 1000 mL and make dissolving, slowly add concentration again and be 36%~38% hydrochloric acid 10mL, shake up, promptly.
Result of the test: in three batches of test sample chromatographs, with the corresponding position of control medicinal material chromatograph on, show the principal spot of same color; And in the negative control chromatograph, with the corresponding position of control medicinal material chromatograph on, display dot not.
Conclusion: this method specificity is strong, good reproducibility, can be used for the discriminating of Radix Rehmanniae Preparata in this concentrated pill.
(3) the methodology checking of Fructus Ligustri Lucidi discriminating:
The preparation of control medicinal material solution: get Fructus Ligustri Lucidi control medicinal material 0.5g, add chloroform 30mL, supersound process 20 minutes filters, and filtrate evaporate to dryness, residue add methanol 1mL, make dissolving at 25~30 ℃, in contrast medical material solution.
The preparation of reference substance solution: even up pier fruit acid reference substance, add methanol, be mixed with the solution that every 1mL contains 1mg at 25~30 ℃, in contrast product solution.
The preparation of need testing solution: get the powder 10g that this product grinds, add chloroform 30mL, supersound process 20 minutes filters, and filtrate evaporate to dryness, residue add methanol 1mL, make dissolving at 25~30 ℃, as need testing solution.Method prepares the need testing solution of three batches of concentrated pills according to this.
The preparation of negative control solution: the negative control sample that lacks Fructus Ligustri Lucidi in embodiment 3 concentrated pills prescription ratio and prepared.Get the powder 10g that the negative control sample grinds, add chloroform 30mL, supersound process 20 minutes filters, and filtrate evaporate to dryness, residue add methanol 1mL, make dissolving at 25~30 ℃, as negative control solution.
Algoscopy: according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B) test, at 25~30 ℃, draw need testing solution each 15 μ L, control medicinal material solution 2 μ L, the reference substance solution 5 μ L of negative control solution and three batches of different batches concentrated pills, putting respectively on same silica gel g thin-layer plate, is developing solvent with chloroform-methanol-formic acid (25:1:1), launches, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to speckle colour developing at 105 ℃.
Result of the test: in three batches of test sample chromatographs, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, show the principal spot of same color; And in the negative control chromatograph, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, display dot not.
Conclusion: this method specificity is strong, good reproducibility, can be used for the discriminating of Fructus Ligustri Lucidi in this concentrated pill.
(4) the methodology checking of Radix Trichosanthis discriminating:
The preparation of control medicinal material solution: get Radix Trichosanthis control medicinal material 2g, add 50% ethanol 20mL, supersound process 30 minutes filters, and gets filtrate medical material solution in contrast.
The preparation of reference substance solution: get the Citruline reference substance, add 50% ethanol, make the solution that every 1mL contains 1mg, in contrast product solution.
The preparation of need testing solution: get the powder 12g that this product grinds, add 70% ethanol 50mL, supersound process 20 minutes filters, and the filtrate evaporate to dryness adds 50% ethanol 5mL and makes dissolving, filters, and gets filtrate as need testing solution.Method prepares the need testing solution of three batches of concentrated pills according to this.
The preparation of negative control solution: the negative control sample that lacks Radix Trichosanthis in embodiment 3 concentrated pills prescription ratio and prepared.Get the powder 12g that the negative control sample grinds, add 70% ethanol 50mL, supersound process 20 minutes filters, and the filtrate evaporate to dryness adds 50% ethanol 5mL and makes dissolving, filters, and gets filtrate as need testing solution.
Algoscopy: according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B) test, draw each 5 μ L of need testing solution, control medicinal material solution and each 2 μ L of reference substance solution of negative control solution and three batches of different batches concentrated pills, put respectively on same silica gel g thin-layer plate, with n-butyl alcohol-glacial acetic acid-water (13: 3:4) be developing solvent, launch, take out, dry, spray is with 2% ethanol solution of ninhydrin, and it is clear to be heated to the speckle colour developing at 105 ℃.
Result of the test: in three batches of test sample chromatographs, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, show the principal spot of same color; And in the negative control chromatograph, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, display dot not.
Conclusion: this method specificity is strong, good reproducibility, can be used for the discriminating of Radix Trichosanthis in this concentrated pill.
Embodiment 8
Prescription and three batches of capsules of method for making preparation by embodiment 4 capsules carry out the methodology checking:
(1) the methodology checking of Fructus Ligustri Lucidi discriminating:
The preparation of control medicinal material solution: get Fructus Ligustri Lucidi control medicinal material 1g, the 100mL that adds methylene chloride, supersound process 10 minutes, filter, filtrate is concentrated into 1mL, evaporate to dryness, residue adds dehydrated alcohol-chloroform by the blended solution 1mL of 3:2, makes dissolving at 20~25 ℃, in contrast medical material solution.
The preparation of need testing solution: get this product content 12g, the 100mL that adds methylene chloride, supersound process 10 minutes, filter, filtrate is concentrated into 1mL, evaporate to dryness, residue adds dehydrated alcohol-chloroform by the blended solution 1mL of 3:2, makes dissolving at 20~25 ℃, as need testing solution.Method prepares the need testing solution of three batches of capsules according to this.
The preparation of negative control solution: the negative control sample that lacks Fructus Ligustri Lucidi in embodiment 4 capsules prescription ratio and prepared.Get negative control sample contents 12g, the 100mL that adds methylene chloride, supersound process 10 minutes filters, and filtrate is concentrated into 1mL, and evaporate to dryness, residue add dehydrated alcohol-chloroform by the blended solution 1mL of 3:2, make dissolving at 20~25 ℃, as negative control solution.
Algoscopy: according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B) test, at 20~25 ℃, draw negative control solution and three crowdes of capsular need testing solutions of different batches each 10 μ L, control medicinal material solution 5 μ L, put respectively on same silica gel g thin-layer plate, with cyclohexane extraction-acetone-ethyl acetate (10:4: 2.5) be developing solvent, launch, take out, dry, spray is with 5% phosphomolybdic acid ethanol solution, and it is clear to be heated to speckle colour developing at 105 ℃.
Result of the test: in three batches of test sample chromatographs, with the corresponding position of control medicinal material chromatograph on, show the principal spot of same color; And in the negative control chromatograph, with the corresponding position of control medicinal material chromatograph on, display dot not.
Conclusion: this method specificity is strong, good reproducibility, can be used for the discriminating of Fructus Ligustri Lucidi in this capsule.
(2) the methodology checking of Radix Trichosanthis discriminating:
The preparation of reference substance solution: get the Citruline reference substance, add 50% ethanol, make the solution that every 1mL contains 1mg, in contrast product solution.
The preparation of need testing solution: get this product content 10g, add 30% ethanol 20mL, supersound process 5 minutes filters, and the filtrate evaporate to dryness adds 50% ethanol 10mL and makes dissolving, filters, and gets filtrate as need testing solution.Method prepares the need testing solution of three batches of capsules according to this.
The preparation of negative control solution: the negative control sample that lacks Radix Trichosanthis in embodiment 4 capsules prescription ratio and prepared.Get negative control sample contents 10g, add 30% ethanol 20mL, supersound process 5 minutes filters, and the filtrate evaporate to dryness adds 50% ethanol 10mL and makes dissolving, filters, and gets filtrate as need testing solution.
Algoscopy: according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B) test, draw negative control solution and three crowdes of capsular need testing solutions of different batches each 10 μ L, reference substance solution 2 μ L, put respectively on same silica gel g thin-layer plate, with n-butyl alcohol-acetone-water (4: 2:1) be developing solvent, launch, take out, dry, spray is with 2% ethanol solution of ninhydrin, and it is clear to be heated to the speckle colour developing at 105 ℃.
Result of the test: in three batches of test sample chromatographs, with the corresponding position of reference substance chromatograph on, show the speckle of same color; And in the negative control chromatograph, with the corresponding position of reference substance chromatograph on, display dot not.
Conclusion: this method specificity is strong, good reproducibility, can be used for the discriminating of Radix Trichosanthis in this capsule.
Claims (8)
1. the discrimination method of the treatment or the Chinese medicine preparation of adjuvant therapy of tumors disease, it is characterized in that: comprise four kinds of Placenta Hominiss, Radix Rehmanniae Preparata, Fructus Ligustri Lucidi, Radix Trichosanthis in the described Chinese medicine preparation, described discrimination method comprise following discrimination method any or any two or wantonly three kinds or all:
(1) discrimination method of Placenta Hominis in the described Chinese medicine preparation is to be the thin layer chromatography discrimination method of contrast with the Placenta Hominis control medicinal material, may further comprise the steps:
1. the preparation of need testing solution: get described Chinese medicine preparation, add chloroform or dichloromethane, supersound process filters, and filtrate concentrates, and makes per 1 mL and contains the solution that the crude drug amount is equivalent to Placenta Hominis 0.2~5g, as need testing solution;
2. the preparation of control medicinal material solution: get the Placenta Hominis control medicinal material, add chloroform or dichloromethane, supersound process filters, and filtrate concentrates, and makes per 1 mL and contains the solution that the crude drug amount is equivalent to Placenta Hominis 0.2~5g, in contrast medical material solution;
3. algoscopy: according to the test of an appendix VI of Chinese Pharmacopoeia version in 2010 B thin layer chromatography, drawing each 2~20 μ L of control medicinal material solution and need testing solution, put respectively on same silica gel g thin-layer plate, is developing solvent with normal hexane-ethyl acetate 9: 1 ~ 9, launch, take out, dry, spray is with 5%~50% perchloric acid solution, it is clear to be heated to speckle colour developing at 60~120 ℃, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the principal spot of same color;
(2) discrimination method of Radix Rehmanniae Preparata in the described Chinese medicine preparation is to be the thin layer chromatography discrimination method of contrast with the Radix Rehmanniae Preparata control medicinal material, may further comprise the steps:
1. the preparation of need testing solution: get described Chinese medicine preparation, add water, warmly make fully molten loosing, be heated to and boil, put cold, centrifugal, get supernatant, concentrate, extract 1~3 time combined ethyl acetate liquid with the ethyl acetate jolting, evaporate to dryness, residue adds methanol makes dissolving, makes per 1 mL and contains the solution that the crude drug amount is equivalent to Radix Rehmanniae Preparata 2~20g, as need testing solution;
2. the preparation of control medicinal material solution: get the Radix Rehmanniae Preparata control medicinal material, decoct with water 1~2 time, filter with absorbent cotton, merging filtrate concentrates, extract 1~3 time with the ethyl acetate jolting, combined ethyl acetate liquid, evaporate to dryness, residue add methanol makes dissolving, make per 1 mL and contain the solution that the crude drug amount is equivalent to Radix Rehmanniae Preparata 2~20g, in contrast medical material solution;
3. algoscopy: according to the test of an appendix VI of Chinese Pharmacopoeia version in 2010 B thin layer chromatography, draw each 2~20 μ L of control medicinal material solution and need testing solution, put respectively on same silica gel g thin-layer plate, with o-Dimethylbenzene-ethyl acetate 1:1 is developing solvent, wherein o-Dimethylbenzene is replaceable is benzene, or toluene, or meta-xylene, or xylol, or o-Dimethylbenzene, meta-xylene, three kinds of mixture of pressing arbitrary proportion of xylol launch, and take out, dry, spray is with 0. 1% 2,4 dinitrophenyl hydrazine alcoholic solution, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the principal spot of same color;
(3) discrimination method of Fructus Ligustri Lucidi in the described Chinese medicine preparation is to be the thin layer chromatography discrimination method of contrast with Fructus Ligustri Lucidi control medicinal material and oleanolic acid reference substance, may further comprise the steps:
1. the preparation of need testing solution: get described Chinese medicine preparation, add chloroform or dichloromethane, extract 1~3 time, merge chloroform liquid or dichloromethane solution, filter, the filtrate evaporate to dryness, residue adds methanol, and perhaps residue adds dehydrated alcohol-chloroform 3:2 mixed solution, at 5~50 ℃, make per 1 mL and contain the solution that the crude drug amount is equivalent to Fructus Ligustri Lucidi 0.2~5g, as need testing solution;
2. the preparation of control medicinal material solution: get the Fructus Ligustri Lucidi control medicinal material, add chloroform or dichloromethane, supersound process, filter, the filtrate evaporate to dryness, residue adds methanol, perhaps residue adds dehydrated alcohol-chloroform 3:2 mixed solution, at 5~50 ℃, make per 1 mL and contain the solution that the crude drug amount is equivalent to Fructus Ligustri Lucidi 0.2~5g, in contrast medical material solution;
3. the preparation of reference substance solution: even up pier fruit acid reference substance, add methanol or 95% ethanol, at 5~50 ℃, be mixed with the solution that every 1mL contains 1mg, in contrast product solution;
4. algoscopy: according to the test of an appendix VI of Chinese Pharmacopoeia version in 2010 B thin layer chromatography, at 5~50 ℃, draw need testing solution, each 2~20 μ L of control medicinal material solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with cyclohexane extraction-acetone-ethyl acetate-glacial acetic acid 7~15: 2~7: 1~5:0~0.5, be developing solvent perhaps with chloroform-methanol-formic acid 20~40:1:0.3~1.1, launch, take out, dry, spray is with 2%~20% ethanol solution of sulfuric acid, perhaps spray with 1%~10% phosphomolybdic acid ethanol solution, it is clear to be heated to speckle colour developing at 60~120 ℃, in the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the principal spot of same color;
(4) discrimination method of Radix Trichosanthis in the described Chinese medicine preparation is to be the thin layer chromatography discrimination method of contrast with Radix Trichosanthis control medicinal material and Citruline reference substance, may further comprise the steps:
1. the preparation of need testing solution: get described Chinese medicine preparation, add 30%~100% ethanol, supersound process filters, the filtrate evaporate to dryness adds 50% ethanol and makes dissolving, filters, get filtrate, make per 1 mL and contain the solution that the crude drug amount is equivalent to Radix Trichosanthis 0.01~1g, as need testing solution;
2. the preparation of control medicinal material solution: get the Radix Trichosanthis control medicinal material, add 50% ethanol, supersound process filters, and gets filtrate, makes per 1 mL and contains the solution that the crude drug amount is equivalent to Radix Trichosanthis 0.01~1g, in contrast medical material solution;
3. the preparation of reference substance solution: get the Citruline reference substance, add 50% ethanol, be mixed with the solution that every 1mL contains 1mg, in contrast product solution;
4. algoscopy: according to the test of an appendix VI of Chinese Pharmacopoeia version in 2010 B thin layer chromatography, draw need testing solution, each 1~20 μ L of control medicinal material solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with n-butyl alcohol-dehydrated alcohol-glacial acetic acid-water 7~15: 0~2:2 ~ 6:1~5, n-butyl alcohol-formic acid-water 15~25: 3:2, n-butyl alcohol-acetone-water 2~5: any among the 2:1 is developing solvent, launch, take out, dry, spray is with 1%~10% ethanol solution of ninhydrin, it is clear to be heated to speckle colour developing at 60~120 ℃, in the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the principal spot of same color.
2. the discrimination method of the Chinese medicine preparation of treatment according to claim 1 or adjuvant therapy of tumors disease, it is characterized in that: described Chinese medicine preparation by Placenta Hominis, the Radix Astragali, Radix Rehmanniae Preparata, Radix Codonopsis, the Rhizoma Atractylodis Macrocephalae, Radix Angelicae Sinensis, Radix Glycyrrhizae, Fructus Ligustri Lucidi, Rhizoma Polygonati, Herba Gueldenstaedtiae or Herba Violae, Radix Trichosanthis ten simply Chinese medicine form, Placenta Hominis is ground into powder, all the other ten flavor medical material solvents extract, and make tablet, capsule, granule, powder, the various suitable dosage forms of pill.
3. the discrimination method of the Chinese medicine preparation of treatment according to claim 2 or adjuvant therapy of tumors disease is characterized in that: the prescription of described Chinese medicine preparation is counted by weight ratio: Placenta Hominis 4~7, the Radix Astragali 20~50, Radix Rehmanniae Preparata 10~30, Radix Codonopsis 5~20, the Rhizoma Atractylodis Macrocephalae 5~20, Radix Angelicae Sinensis 5~15, Radix Glycyrrhizae 5~15, Fructus Ligustri Lucidi 5~20, Rhizoma Polygonati 5~15, Herba Gueldenstaedtiae or Herba Violae 5~20, Radix Trichosanthis 5~20.
4. the discrimination method of the Chinese medicine preparation of treatment according to claim 3 or adjuvant therapy of tumors disease is characterized in that: every gram contains the crude drug amount and is equivalent to Placenta Hominis 0.15~0.18g, Radix Rehmanniae Preparata 0.45~0.55g, Fructus Ligustri Lucidi 0.25~0.35g, Radix Trichosanthis 0.25~0.35g in the described Chinese medicine preparation.
5. the discrimination method of the Chinese medicine preparation of treatment according to claim 1 or adjuvant therapy of tumors disease, it is characterized in that: described 0.1% 2,4-dinitrophenylhydrazine alcoholic solution is: get 2,4-dinitrophenylhydrazine 1g, add dehydrated alcohol 1000 mL and make dissolving, slowly add concentration again and be 36%~38% hydrochloric acid 10mL, shake up, promptly.
6. the discrimination method of the Chinese medicine preparation of treatment according to claim 1 or adjuvant therapy of tumors disease, it is characterized in that: the discrimination method of Fructus Ligustri Lucidi in the described Chinese medicine preparation, before the preparation need testing solution, carry out pretreatment to Chinese medicine preparation, concrete operations are: get described Chinese medicine preparation, add 0.2%~0.5% sodium hydroxide solution, warmly make fully molten loosing, be heated to and boil, put cold, filtrate or supernatant are got in filtration or centrifugal, regulate pH value to 1~2 with hydrochloric acid or sulphuric acid.
7. according to the discrimination method of the Chinese medicine preparation of described treatment of claim 1 ~ 6 or adjuvant therapy of tumors disease, it is characterized in that: described discrimination method comprise following discrimination method any or any two or wantonly three kinds or all:
(1) discrimination method of Placenta Hominis in the described Chinese medicine preparation is to be the thin layer chromatography discrimination method of contrast with the Placenta Hominis control medicinal material, may further comprise the steps:
1. the preparation of need testing solution: get described Chinese medicine preparation 3 ~ 10g, add chloroform or dichloromethane 20~30mL, supersound process 5~45 minutes, filter, concentrated filtrate is made per 1 mL and is contained the solution that the crude drug amount is equivalent to Placenta Hominis 0.2~5g, as need testing solution;
2. the preparation of control medicinal material solution: get Placenta Hominis control medicinal material 0.5 ~ 2g, add chloroform or dichloromethane 20~30mL, supersound process 5~45 minutes, filter, concentrated filtrate is made per 1 mL and is contained the solution that the crude drug amount is equivalent to Placenta Hominis 0.2~5g, in contrast medical material solution;
3. algoscopy: according to the test of an appendix VI of Chinese Pharmacopoeia version in 2010 B thin layer chromatography, drawing each 5~15 μ L of control medicinal material solution and need testing solution, put respectively on same silica gel g thin-layer plate, is developing solvent with normal hexane-ethyl acetate 9: 1 ~ 9, launch, take out, dry, spray perchloric acid solution with 5 ~ 50%, it is clear to be heated to speckle colour developing at 60 ~ 120 ℃, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the principal spot of same color;
(2) discrimination method of Radix Rehmanniae Preparata in the described Chinese medicine preparation is to be the thin layer chromatography discrimination method of contrast with the Radix Rehmanniae Preparata control medicinal material, may further comprise the steps:
1. the preparation of need testing solution: get described Chinese medicine preparation 6~15g, add water 100~200mL, warmly make fully molten loosing, be heated to and boil, put cold, centrifugal, get supernatant, be concentrated into 30~100mL, extract 1~3 time with the ethyl acetate jolting, each 20~30mL, combined ethyl acetate liquid, evaporate to dryness, residue add methanol makes dissolving, make per 1 mL and contain the solution that the crude drug amount is equivalent to Radix Rehmanniae Preparata 2~20g, as need testing solution;
2. the preparation of control medicinal material solution: get Radix Rehmanniae Preparata control medicinal material 2~10g, decoct with water 1 ~ 2 time, add water 50~150mL at every turn, the each decoction 0.5~2 hour filters merging filtrate with absorbent cotton, be concentrated into 30~100mL, extract 1~3 time each 20~30mL, combined ethyl acetate liquid with the ethyl acetate jolting, evaporate to dryness, residue adds methanol makes dissolving, makes per 1 mL and contains the solution that the crude drug amount is equivalent to Radix Rehmanniae Preparata 2~20g, in contrast medical material solution;
3. algoscopy: according to the test of an appendix VI of Chinese Pharmacopoeia version in 2010 B thin layer chromatography, draw each 5~15 μ L of control medicinal material solution and need testing solution, put respectively on same silica gel g thin-layer plate, with o-Dimethylbenzene-ethyl acetate 1:1 is developing solvent, wherein o-Dimethylbenzene is replaceable is benzene, or toluene, or meta-xylene, or xylol, or o-Dimethylbenzene, meta-xylene, three kinds of mixture of pressing arbitrary proportion of xylol launch, and take out, dry, spray is with 0.1% 2,4 dinitrophenyl hydrazine alcoholic solution, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the principal spot of same color;
(3) discrimination method of Fructus Ligustri Lucidi in the described Chinese medicine preparation is to be the thin layer chromatography discrimination method of contrast with Fructus Ligustri Lucidi control medicinal material and oleanolic acid reference substance, may further comprise the steps:
1. the preparation of need testing solution: get described Chinese medicine preparation 1.5~12g, add chloroform or dichloromethane 20~100mL, supersound process 5~45 minutes, filter, filtrate evaporate to dryness, residue add methanol or add dehydrated alcohol-chloroform by the blended solution of 3:2, make dissolving at 10~50 ℃, make per 1 mL and contain the solution that the crude drug amount is equivalent to Fructus Ligustri Lucidi 0.2~5g, as need testing solution;
2. the preparation of control medicinal material solution: get Fructus Ligustri Lucidi control medicinal material 0.5g, add chloroform or dichloromethane 20mL, supersound process 5 ~ 45 minutes, filter, filtrate evaporate to dryness, residue add methanol or add dehydrated alcohol-chloroform by the blended solution of 3:2, make dissolving at 10~50 ℃, make per 1 mL and contain the solution that the crude drug amount is equivalent to Fructus Ligustri Lucidi 0.2~5g, in contrast medical material solution;
3. the preparation of reference substance solution: even up pier fruit acid reference substance, at 10~50 ℃, add methanol or 95% ethanol is mixed with the solution that every 1mL contains 1mg, in contrast product solution;
4. algoscopy: according to the test of an appendix VI of Chinese Pharmacopoeia version in 2010 B thin layer chromatography, at 10~50 ℃, draw need testing solution 5~15 μ L, each 2~5 μ L of control medicinal material solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with cyclohexane extraction-acetone-ethyl acetate-glacial acetic acid 7~15: 2~7: 1~5:0~0.5, be developing solvent perhaps with chloroform-methanol-formic acid 20~40:1:0.3~1.1, launch, take out, dry, spray is with 2 ~ 20% ethanol solution of sulfuric acid, perhaps spray with 1 ~ 10% phosphomolybdic acid ethanol solution, it is clear to be heated to speckle colour developing at 60~120 ℃, in the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the principal spot of same color;
(4) discrimination method of Radix Trichosanthis in the described Chinese medicine preparation is to be the thin layer chromatography discrimination method of contrast with Radix Trichosanthis control medicinal material and Citruline reference substance, may further comprise the steps:
1. the preparation of need testing solution: get described Chinese medicine preparation 6~12g, add 30%~100% ethanol, supersound process 5~45 minutes, filter, the filtrate evaporate to dryness adds 50% ethanol and makes dissolving, filter, make per 1 mL and contain the solution that the crude drug amount is equivalent to Radix Trichosanthis 0.01~1g, as need testing solution;
2. the preparation of control medicinal material solution: get Radix Trichosanthis control medicinal material 2g, add 50% ethanol, supersound process 5 ~ 45 minutes filters, and makes per 1 mL and contains the solution that the crude drug amount is equivalent to Radix Trichosanthis 0.01~1g, in contrast medical material solution;
3. the preparation of reference substance solution: get the Citruline reference substance, add 50% ethanol and be mixed with the solution that every 1mL contains 1mg, in contrast product solution;
4. algoscopy: according to the test of an appendix VI of Chinese Pharmacopoeia version in 2010 B thin layer chromatography, draw need testing solution 5~15 μ L, each 1~5 μ L of control medicinal material solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with n-butyl alcohol-dehydrated alcohol-glacial acetic acid-water 7~15: 0~2:2 ~ 6:1~5, n-butyl alcohol-formic acid-water 15~25: 3:2, n-butyl alcohol-acetone-water 2~5: any among the 2:1 is developing solvent, launch, take out, dry, spray is with 1 ~ 10% ethanol solution of ninhydrin, it is clear to be heated to speckle colour developing at 60 ~ 120 ℃, in the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the principal spot of same color.
8. the discrimination method of the Chinese medicine preparation of treatment according to claim 7 or adjuvant therapy of tumors disease is characterized in that: described discrimination method comprise following discrimination method any or any two or wantonly three kinds or all:
(1) discrimination method of Placenta Hominis in the described Chinese medicine preparation may further comprise the steps:
Get described Chinese medicine preparation 6g, add chloroform 20mL, supersound process 30 minutes filters, and filtrate is concentrated into 1mL, as need testing solution; Other gets Placenta Hominis control medicinal material 1g, shines medical material solution in pairs with legal system; Test according to an appendix VI of Chinese Pharmacopoeia version in 2010 B thin layer chromatography, draw each 10 μ L of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with normal hexane-ethyl acetate is developing solvent at 9: 5, launch, take out, dry, spray is with 20% perchloric acid solution, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the principal spot of same color;
(2) discrimination method of Radix Rehmanniae Preparata in the described Chinese medicine preparation may further comprise the steps:
Get described Chinese medicine preparation 10g, add water 150mL, warmly make fully molten loosing, be heated to and boil, put cold, centrifugal, get supernatant, be concentrated into 100mL, extract 2 times each 30mL with the ethyl acetate jolting, combined ethyl acetate liquid, evaporate to dryness, residue add methanol 0.5mL makes dissolving, as need testing solution; Other gets Radix Rehmanniae Preparata control medicinal material 5g, decocts with water 2 times, adds water 80mL at every turn, 2 hours for the first time, 1 hour for the second time, filter with absorbent cotton, merging filtrate, be concentrated into 100mL, extract 2 times, each 30mL with the ethyl acetate jolting, combined ethyl acetate liquid, evaporate to dryness, residue add methanol 0.5mL makes dissolving, in contrast medical material solution; Test according to an appendix VI of Chinese Pharmacopoeia version in 2010 B thin layer chromatography, 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 o-Dimethylbenzene-ethyl acetate is developing solvent at 1: 1, launch, take out, dry, spray is with 0.1% 2,4 dinitrophenyl hydrazine alcoholic solution; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the principal spot of same color;
(3) discrimination method of Fructus Ligustri Lucidi in the described Chinese medicine preparation may further comprise the steps:
Get described Chinese medicine preparation 6g, add chloroform 20mL, supersound process 30 minutes filters, and filtrate evaporate to dryness, residue add methanol 1mL, make dissolving at 35~40 ℃, as need testing solution; Other gets Fructus Ligustri Lucidi control medicinal material 0.5g, shines medical material solution in pairs with legal system; Even up pier fruit acid reference substance again, add methanol and be mixed with the solution that every 1mL contains 1mg, in contrast product solution at 35~40 ℃; Test according to an appendix VI of Chinese Pharmacopoeia version in 2010 B thin layer chromatography, at 35~40 ℃, draw each 5 μ L of need testing solution 10 ~ 15 μ L, control medicinal material solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with cyclohexane extraction-acetone-ethyl acetate-glacial acetic acid 12.5: 5: 3:0.1 was developing solvent, launches, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to speckle colour developing at 110 ℃; In the test sample chromatograph, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, show the principal spot of same color;
(4) discrimination method of Radix Trichosanthis in the described Chinese medicine preparation may further comprise the steps:
Get described Chinese medicine preparation 10g, add 50% ethanol 30mL, supersound process 30 minutes filters, and the filtrate evaporate to dryness adds 50% ethanol 10mL and makes dissolving, filters, and gets filtrate as need testing solution; Other gets Radix Trichosanthis control medicinal material 2g, adds 50% ethanol 20mL, and supersound process 30 minutes filters, and gets filtrate medical material solution in contrast; Get the Citruline reference substance again, add 50% ethanol and make the solution that every 1mL contains 1mg, in contrast product solution; Test according to an appendix VI of Chinese Pharmacopoeia version in 2010 B thin layer chromatography, draw each 2 μ L of need testing solution 10 μ L, control medicinal material solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with n-butyl alcohol-dehydrated alcohol-glacial acetic acid-water 8: 2:4:3 is developing solvent, launch, take out, dry, spray is with 2% ethanol solution of ninhydrin, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, show the principal spot of same color.
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CN105181846A (en) * | 2015-09-28 | 2015-12-23 | 成都维恒医药科技有限公司 | Method for identifying human placenta |
CN105675791A (en) * | 2016-02-05 | 2016-06-15 | 四川德成动物保健品有限公司 | Antipyretic-and-antitoxic-powder pretreatment method for detecting rehmannia in thin-layer chromatography mode |
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CN101804155A (en) * | 2010-04-22 | 2010-08-18 | 王志敏 | Medicine used for treating qi and blood deficiency caused by radiotherapy and chemotherapy of tumor and postoperative body weakness |
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CN101804155A (en) * | 2010-04-22 | 2010-08-18 | 王志敏 | Medicine used for treating qi and blood deficiency caused by radiotherapy and chemotherapy of tumor and postoperative body weakness |
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CN105181846A (en) * | 2015-09-28 | 2015-12-23 | 成都维恒医药科技有限公司 | Method for identifying human placenta |
CN105675791A (en) * | 2016-02-05 | 2016-06-15 | 四川德成动物保健品有限公司 | Antipyretic-and-antitoxic-powder pretreatment method for detecting rehmannia in thin-layer chromatography mode |
CN111024876A (en) * | 2019-12-25 | 2020-04-17 | 石家庄平安医院有限公司 | Gradient full-information thin-layer identification method for glossy privet fruit medicinal materials |
CN111024876B (en) * | 2019-12-25 | 2021-08-10 | 石家庄平安医院有限公司 | Gradient full-information thin-layer identification method for glossy privet fruit medicinal materials |
CN113514597A (en) * | 2021-07-14 | 2021-10-19 | 鲁南厚普制药有限公司 | Thin-layer chromatography identification method for Gastrodia elata dizziness relieving granules |
CN113514597B (en) * | 2021-07-14 | 2024-05-31 | 鲁南厚普制药有限公司 | Thin-layer chromatography identification method of gastrodia tuber dizziness relieving granule |
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