CN108760945A - A kind of detection method of Qijiaoshenbai capsule - Google Patents
A kind of detection method of Qijiaoshenbai capsule Download PDFInfo
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- 239000002775 capsule Substances 0.000 title claims abstract description 46
- 238000001514 detection method Methods 0.000 title claims abstract description 38
- ZSBXGIUJOOQZMP-UHFFFAOYSA-N Isomatrine Natural products C1CCC2CN3C(=O)CCCC3C3C2N1CCC3 ZSBXGIUJOOQZMP-UHFFFAOYSA-N 0.000 claims abstract description 75
- 239000000706 filtrate Substances 0.000 claims abstract description 70
- 241001247821 Ziziphus Species 0.000 claims abstract description 66
- ZSBXGIUJOOQZMP-JLNYLFASSA-N Matrine Chemical compound C1CC[C@H]2CN3C(=O)CCC[C@@H]3[C@@H]3[C@H]2N1CCC3 ZSBXGIUJOOQZMP-JLNYLFASSA-N 0.000 claims abstract description 65
- 229930014456 matrine Natural products 0.000 claims abstract description 65
- 238000000034 method Methods 0.000 claims abstract description 61
- 239000003814 drug Substances 0.000 claims abstract description 45
- 238000001914 filtration Methods 0.000 claims abstract description 44
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 44
- 229940079593 drug Drugs 0.000 claims abstract description 41
- TZJALUIVHRYQQB-XFDQAQKOSA-N Icariin Natural products O(C)c1ccc(C2=C(O[C@H]3[C@@H](O)[C@H](O)[C@@H](O)[C@H](C)O3)C(=O)c3c(O)cc(O[C@H]4[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O4)c(C/C=C(\C)/C)c3O2)cc1 TZJALUIVHRYQQB-XFDQAQKOSA-N 0.000 claims abstract description 28
- TZJALUIVHRYQQB-XLRXWWTNSA-N icariin Chemical compound C1=CC(OC)=CC=C1C1=C(O[C@H]2[C@@H]([C@H](O)[C@@H](O)[C@H](C)O2)O)C(=O)C2=C(O)C=C(O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O3)O)C(CC=C(C)C)=C2O1 TZJALUIVHRYQQB-XLRXWWTNSA-N 0.000 claims abstract description 28
- TZJALUIVHRYQQB-UHFFFAOYSA-N icariine Natural products C1=CC(OC)=CC=C1C1=C(OC2C(C(O)C(O)C(C)O2)O)C(=O)C2=C(O)C=C(OC3C(C(O)C(O)C(CO)O3)O)C(CC=C(C)C)=C2O1 TZJALUIVHRYQQB-UHFFFAOYSA-N 0.000 claims abstract description 28
- 239000009636 Huang Qi Substances 0.000 claims abstract description 19
- 238000003556 assay Methods 0.000 claims abstract description 16
- 108010010803 Gelatin Proteins 0.000 claims abstract description 13
- 239000008273 gelatin Substances 0.000 claims abstract description 13
- 229920000159 gelatin Polymers 0.000 claims abstract description 13
- 235000019322 gelatine Nutrition 0.000 claims abstract description 13
- 235000011852 gelatine desserts Nutrition 0.000 claims abstract description 13
- 239000000843 powder Substances 0.000 claims abstract description 12
- 235000019640 taste Nutrition 0.000 claims abstract description 9
- -1 mixing is added Substances 0.000 claims abstract description 7
- 238000012850 discrimination method Methods 0.000 claims abstract description 6
- 239000002245 particle Substances 0.000 claims abstract description 6
- 241000522169 Lespedeza Species 0.000 claims abstract description 5
- 238000004097 X-ray Buerger Methods 0.000 claims abstract description 5
- 238000002156 mixing Methods 0.000 claims abstract description 5
- 238000003756 stirring Methods 0.000 claims abstract description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 267
- 239000000243 solution Substances 0.000 claims description 145
- 238000004587 chromatography analysis Methods 0.000 claims description 106
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 104
- 239000013558 reference substance Substances 0.000 claims description 104
- 239000012085 test solution Substances 0.000 claims description 89
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 84
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical class CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 80
- 238000012360 testing method Methods 0.000 claims description 76
- 239000007788 liquid Substances 0.000 claims description 75
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 69
- 239000000523 sample Substances 0.000 claims description 62
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 61
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 58
- 239000000463 material Substances 0.000 claims description 56
- 235000019441 ethanol Nutrition 0.000 claims description 43
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 40
- 238000000605 extraction Methods 0.000 claims description 38
- 238000010992 reflux Methods 0.000 claims description 37
- 239000000741 silica gel Substances 0.000 claims description 33
- 229910002027 silica gel Inorganic materials 0.000 claims description 33
- 239000002904 solvent Substances 0.000 claims description 31
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims description 30
- 239000008187 granular material Substances 0.000 claims description 30
- 238000002360 preparation method Methods 0.000 claims description 30
- 239000000047 product Substances 0.000 claims description 26
- 238000005406 washing Methods 0.000 claims description 22
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims description 21
- 239000007921 spray Substances 0.000 claims description 21
- 239000003208 petroleum Substances 0.000 claims description 20
- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 16
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 15
- 238000002474 experimental method Methods 0.000 claims description 15
- 229920006395 saturated elastomer Polymers 0.000 claims description 15
- 229960000583 acetic acid Drugs 0.000 claims description 14
- 239000012362 glacial acetic acid Substances 0.000 claims description 14
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 14
- 239000000377 silicon dioxide Substances 0.000 claims description 14
- 239000000945 filler Substances 0.000 claims description 13
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims description 13
- 238000010438 heat treatment Methods 0.000 claims description 12
- 239000006210 lotion Substances 0.000 claims description 12
- 238000002604 ultrasonography Methods 0.000 claims description 11
- SIHHLZPXQLFPMC-UHFFFAOYSA-N chloroform;methanol;hydrate Chemical compound O.OC.ClC(Cl)Cl SIHHLZPXQLFPMC-UHFFFAOYSA-N 0.000 claims description 10
- 238000010828 elution Methods 0.000 claims description 10
- ZSBXGIUJOOQZMP-BHPKHCPMSA-N sophoridine Chemical compound C1CC[C@@H]2CN3C(=O)CCC[C@@H]3[C@@H]3[C@H]2N1CCC3 ZSBXGIUJOOQZMP-BHPKHCPMSA-N 0.000 claims description 10
- 239000000284 extract Substances 0.000 claims description 9
- 238000009736 wetting Methods 0.000 claims description 9
- 238000004128 high performance liquid chromatography Methods 0.000 claims description 8
- 239000012488 sample solution Substances 0.000 claims description 8
- 239000012467 final product Substances 0.000 claims description 7
- 239000006071 cream Substances 0.000 claims description 6
- 238000011161 development Methods 0.000 claims description 6
- 239000008186 active pharmaceutical agent Substances 0.000 claims description 5
- 229910021529 ammonia Inorganic materials 0.000 claims description 5
- QMNWISYXSJWHRY-YLNUDOOFSA-N astragaloside IV Chemical compound O1[C@H](C(C)(O)C)CC[C@]1(C)[C@@H]1[C@@]2(C)CC[C@]34C[C@]4(CC[C@H](O[C@H]4[C@@H]([C@@H](O)[C@H](O)CO4)O)C4(C)C)[C@H]4[C@@H](O[C@H]4[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O4)O)C[C@H]3[C@]2(C)C[C@@H]1O QMNWISYXSJWHRY-YLNUDOOFSA-N 0.000 claims description 5
- QMNWISYXSJWHRY-BCBPIKMJSA-N astragaloside IV Natural products CC(C)(O)[C@@H]1CC[C@@](C)(O1)[C@H]2[C@@H](O)C[C@@]3(C)[C@@H]4C[C@H](O[C@@H]5O[C@H](CO)[C@H](O)[C@@H](O)[C@H]5O)[C@H]6C(C)(C)[C@H](CC[C@@]67C[C@@]47CC[C@]23C)O[C@@H]8OC[C@@H](O)[C@H](O)[C@H]8O QMNWISYXSJWHRY-BCBPIKMJSA-N 0.000 claims description 5
- PFKIBRPYVNVMRU-UHFFFAOYSA-N cyclosieversioside F Natural products CC(C)(O)C1COC(C)(C1)C2C(O)CC3(C)C4CC(OC5OC(CO)C(O)C(O)C5O)C6C(C)(C)C(CCC67CC47CCC23C)OC8OCC(O)C(O)C8O PFKIBRPYVNVMRU-UHFFFAOYSA-N 0.000 claims description 5
- 229940088679 drug related substance Drugs 0.000 claims description 5
- OAYLNYINCPYISS-UHFFFAOYSA-N ethyl acetate;hexane Chemical compound CCCCCC.CCOC(C)=O OAYLNYINCPYISS-UHFFFAOYSA-N 0.000 claims description 5
- 238000000079 presaturation Methods 0.000 claims description 5
- 239000006228 supernatant Substances 0.000 claims description 5
- 241000238370 Sepia Species 0.000 claims description 3
- 239000007902 hard capsule Substances 0.000 claims description 3
- 239000002893 slag Substances 0.000 claims description 3
- 238000010521 absorption reaction Methods 0.000 claims 2
- XYIBRDXRRQCHLP-UHFFFAOYSA-N ethyl acetoacetate Chemical compound CCOC(=O)CC(C)=O XYIBRDXRRQCHLP-UHFFFAOYSA-N 0.000 claims 1
- 239000012071 phase Substances 0.000 description 27
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 12
- 238000012545 processing Methods 0.000 description 12
- 238000004088 simulation Methods 0.000 description 10
- 238000004458 analytical method Methods 0.000 description 9
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 8
- 241000246044 Sophora flavescens Species 0.000 description 7
- 239000007864 aqueous solution Substances 0.000 description 5
- 239000003795 chemical substances by application Substances 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 239000003513 alkali Substances 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- 238000005259 measurement Methods 0.000 description 4
- 238000005070 sampling Methods 0.000 description 4
- 238000004809 thin layer chromatography Methods 0.000 description 4
- 238000012546 transfer Methods 0.000 description 4
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonium chloride Substances [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 3
- 241000446313 Lamella Species 0.000 description 3
- ONBIUAZBGHXJDM-UHFFFAOYSA-J bismuth;potassium;tetraiodide Chemical compound [K+].[I-].[I-].[I-].[I-].[Bi+3] ONBIUAZBGHXJDM-UHFFFAOYSA-J 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000003292 glue Substances 0.000 description 3
- 239000004615 ingredient Substances 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- 238000012417 linear regression Methods 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- 238000000527 sonication Methods 0.000 description 3
- XVPBINOPNYFXID-JARXUMMXSA-N 85u4c366qs Chemical compound C([C@@H]1CCC[N@+]2(CCC[C@H]3[C@@H]21)[O-])N1[C@@H]3CCCC1=O XVPBINOPNYFXID-JARXUMMXSA-N 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 241001494479 Pecora Species 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- PJANXHGTPQOBST-VAWYXSNFSA-N Stilbene Natural products C=1C=CC=CC=1/C=C/C1=CC=CC=C1 PJANXHGTPQOBST-VAWYXSNFSA-N 0.000 description 2
- 238000009835 boiling Methods 0.000 description 2
- 229940126678 chinese medicines Drugs 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 229930182470 glycoside Natural products 0.000 description 2
- 150000002338 glycosides Chemical class 0.000 description 2
- 238000011835 investigation Methods 0.000 description 2
- 239000007791 liquid phase Substances 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 2
- 229930015582 oxymatrine Natural products 0.000 description 2
- PJANXHGTPQOBST-UHFFFAOYSA-N stilbene Chemical compound C=1C=CC=CC=1C=CC1=CC=CC=C1 PJANXHGTPQOBST-UHFFFAOYSA-N 0.000 description 2
- 235000021286 stilbenes Nutrition 0.000 description 2
- 241000208340 Araliaceae Species 0.000 description 1
- 241000512000 Asclepias curassavica Species 0.000 description 1
- 208000000059 Dyspnea Diseases 0.000 description 1
- 206010013975 Dyspnoeas Diseases 0.000 description 1
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 description 1
- 235000003140 Panax quinquefolius Nutrition 0.000 description 1
- 150000001343 alkyl silanes Chemical group 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 208000002173 dizziness Diseases 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000008434 ginseng Nutrition 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000005304 joining Methods 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 201000002364 leukopenia Diseases 0.000 description 1
- 231100001022 leukopenia Toxicity 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- 230000003252 repetitive effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 208000013220 shortness of breath Diseases 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 238000004611 spectroscopical analysis Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 230000002269 spontaneous effect Effects 0.000 description 1
- 230000035900 sweating Effects 0.000 description 1
- 230000001550 time effect Effects 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 238000010200 validation analysis Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/90—Plate chromatography, e.g. thin layer or paper chromatography
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Medicinal Preparation (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
A kind of detection method of Qijiaoshenbai capsule, the drug are prepared by the following method by jujube, donkey-hide gelatin, buerger lespedeza root, Herba Epimedii, kuh-seng, Radix Astragali and Radix Angelicae Sinensis：Above seven taste, in addition to donkey-hide gelatin, Radix Angelicae Sinensis is ground into fine powder；The five tastes such as remaining jujube add water to cook three times, 2 hours for the first time, second 1.5 hours, third time 1 hour, collecting decoction, filtration, filtrate are added the donkey-hide gelatin after molten, stir evenly, it is concentrated into the thick paste that relative density is 1.30 at 75-85 DEG C, above-mentioned Radix Angelicae Sinensis fine powder, mixing is added, particle is made, it is dry, be packed into capsule to get；The detection method includes discrimination method and content assaying method；The discrimination method includes thin layer discriminating, the thin layer discriminating of Radix Angelicae Sinensis, the thin layer discriminating of kuh-seng, the thin layer discriminating of Radix Astragali and/or the thin layer discriminating of Herba Epimedii to jujube；The content assaying method includes the assay to matrine and/or icariin.
Description
Technical field
The present invention relates to a kind of detection methods of Qijiaoshenbai capsule, belong to the field of drug technology；
Background technology
Qijiaoshenbai capsule be a seedling doctor Chinese medicine complementation ethnic drug, mainly by jujube, donkey-hide gelatin, buerger lespedeza root, Herba Epimedii,
The compositions such as kuh-seng, Radix Astragali, Radix Angelicae Sinensis have effects that enriching the blood and tonifying qi, are clinically used to treat the dizzy eye caused by deficiency of vital energy and blood
Flower, shortness of breath and fatigue, spontaneous sweating and leukopenia etc..
Currently, the standard that Qijiaoshenbai capsule executes is State Food and Drug Administration's standard (number WS-10026
(ZD-0026-2002-2012Z)) Qualitive test only, in the standard, is carried out to Radix Angelicae Sinensis, Radix Astragali, kuh-seng, Herba Epimedii, to excessive sheep
Leaves of pulse plants glycosides carries out quantitative check.But the compound Chinese medicinal preparation that Qijiaoshenbai capsule is formed as 7 taste Chinese medicines, prescription element are multiple
It is miscellaneous, according to current quality standard, it is that cannot preferably control its quality standard, promotes the inherent quality of product.
Invention content
The object of the present invention is to provide a kind of detection methods of Qijiaoshenbai capsule, and the detection method is accurate, specially
Attribute is strong, can be as the method for quality control of Qijiaoshenbai capsule capsule.The present invention in Qijiaoshenbai capsule to jujube, when
Return, the carry out thin layer discriminating of kuh-seng, Radix Astragali and Herba Epimedii；Assay is carried out to matrine and/or icariin, increases jujube
Indentification by TLC；The amount for measuring matrine in Sophora flavescens, to establish effectively reliable quality standard, in capable of accurately reflecting
Medicine total quality provides data reference.The treatment of the product can preferably be controlled, it is ensured that its clinical efficacy.
In order to solve the above technical problems, the present invention adopts the following technical scheme that realization：A kind of detection of Qijiaoshenbai capsule
Method, the drug is by 240-260 parts of jujube, 240-250 parts of donkey-hide gelatin, 365-385 parts of buerger lespedeza root, 365-385 parts of Herba Epimedii, hardship
240-260 parts of 240-260 parts of ginseng, 740-760 parts of Radix Astragali and Radix Angelicae Sinensis are prepared by the following method：Above seven taste removes donkey-hide gelatin
Outside, Radix Angelicae Sinensis is ground into fine powder；The five tastes such as remaining jujube add water to cook three times, 2 hours for the first time, second 1.5 hours, third
Secondary 1 hour, collecting decoction filtered, and the donkey-hide gelatin after molten is added in filtrate, stirs evenly, and it is 1.30 to be concentrated into relative density at 75-85 DEG C
Thick paste, above-mentioned Radix Angelicae Sinensis fine powder is added, particle is made in mixing, dry, be packed into capsule to get；The detection method includes differentiating
Method and content assaying method；The discrimination method include the thin layer of jujube is differentiated, the thin layer of Radix Angelicae Sinensis differentiates, the thin layer of kuh-seng
Differentiate, the thin layer of Radix Astragali differentiates and/or the thin layer of Herba Epimedii differentiates；The content assaying method includes to matrine and/or excessive
The assay of sheep leaves of pulse plants glycosides.
In the detection method of aforementioned Qijiaoshenbai capsule, the thin layer discriminating of the jujube is：Drug granule content 3g is taken,
Add ethyl acetate 30ml, be ultrasonically treated 30 minutes, filtering is evaporated filtrate, residue adds methanol 0.5ml to make dissolving, as test sample
Solution；Jujube control medicinal material 2g separately is taken, adds water 50ml, is heated to reflux 2 hours, filters while hot, filtrate volatilizes moisture and jujube is made
Thick paste, cream add ethyl acetate 30ml, are ultrasonically treated 30 minutes, and filtering, filtrate is evaporated, and adds methanol 1ml to make dissolving, as a contrast medicine
Material solution draws 5 μ l of test solution and 10 μ l of control medicinal material solution, is put respectively on same silica gel H lamellae, with toluene-
Ethyl acetate-glacial acetic acid=60:6:0.05 is solvent, is unfolded, and takes out, dries, and is sprayed with 10% ethanol solution of sulfuric acid, 105
It is clear DEG C to be heated to spot development, it is to be inspected under 365nm ultraviolet lamps to set wavelength, in test sample chromatography, with reference substance chromatography
On corresponding position, the fluorescence spot of same color is shown.
In the detection method of aforementioned Qijiaoshenbai capsule, the thin-layer identification method of the Radix Angelicae Sinensis is：Take drug granule content
Object 5g adds n-hexane 20ml, is ultrasonically treated 20 minutes, and filtration, filtrate is waved to 1ml, as test solution；Separately Radix Angelicae Sinensis is taken to compare
Medicinal material 0.5g is made in the same way of control medicinal material solution；It is tested according to thin-layered chromatography (one VI B of annex of Chinese Pharmacopoeia version in 2010),
Each 5 μ l of above two solution are drawn, are put respectively on same silica gel g thin-layer plate, with n-hexane-ethyl acetate=9:1 is expansion
Agent is unfolded, and takes out, dries, and it is to be inspected under 365nm ultraviolet lamps to set wavelength, in test sample chromatography, is that control medicinal material chromatography is corresponding
Position on, the fluorescence spot of first same color；
The thin-layer identification method of the kuh-seng is：Drug granule content 1g is taken, ethyl alcohol 30ml is added, is heated to reflux 30 points
Clock is let cool, and filtration, filtrate is evaporated residue and water 20ml is added to make dissolving, is filtered, and filtrate is set in separatory funnel, and a concentration of 25%- is added
28% strong ammonia solution 0.5ml is merged chloroform liquid, is evaporated, residue adds with chloroform shaking extraction 2 times, each 10ml
Absolute ethyl alcohol 1ml makes dissolving, as test solution；Matrine and Sophoridine reference substance separately are taken, respectively plus absolute ethyl alcohol is made
Per solution of the 1ml containing 1mg, product solution, is tried according to thin-layered chromatography (one VI B of annex of Chinese Pharmacopoeia version in 2010) as a contrast
It tests, draws above-mentioned each 5 μ l of three kinds of solution, put respectively on same silica gel g thin-layer plate, it is dense with cyclohexane-acetone-ethyl acetate-
Ammonia solution=2:3:4:0.5 is solvent, is set in the pre-saturated exhibition cylinder of ammonia steam, presaturation 30 minutes, is unfolded, and takes out, dries,
Spray is with dilute bismuth potassium iodide test solution；In test sample chromatography, it is on the corresponding position of reference substance chromatography, shows the spot of same color；
The thin-layer identification method of the Radix Astragali is：Drug granule content 5g is taken, 60-90 DEG C of petroleum ether 30ml is added, is surpassed
Sonication 20 minutes, filtration, the dregs of a decoction wave most petroleum ether, add methanol 50ml, are heated to reflux 30 minutes, filter, a small amount of first of filter residue
Alcohol washs, and washing lotion merges with filtrate, is evaporated, and residue adds water 10ml, heating to make dissolving, let cool, set in centrifuge tube and centrifuge 10 minutes,
Supernatant is taken, with chloroform shaking extraction 2 times, each 15ml discards chloroform liquid, is shaken to water saturated n-butanol
Extraction 3 times adds 10ml for the first time, and 10ml, third time plus 5ml, merging n-butanol liquid is added to ammoniate test solution washing 3 times, often for the second time
Secondary 15ml, discards ammoniacal liquor；N-butanol liquid is washed to neutrality to what n-butanol was saturated, discards aqueous, and n-butanol liquid is evaporated, residue
Methanol 1ml is added to make dissolving, as test solution；Astragaloside IV reference substance separately is taken, adds methanol that solution of every 1ml containing 1mg is made,
Product solution as a contrast；It is tested according to thin-layered chromatography (one VI B of annex of Chinese Pharmacopoeia version in 2010), draws test solution 10
μ l, 5 μ l of reference substance solution are put respectively on same silica gel g thin-layer plate, with chloroform-methanol-water=13:7:2,10 DEG C with
Lower layer's solution of lower placement is solvent, is unfolded, and takes out, dries, and sprays with 10% ethanol solution of sulfuric acid, spot is heated at 105 DEG C
Point colour developing is clear；In test sample chromatography, on position corresponding with reference substance chromatography, the spot of same color is shown；Wavelength is set again
To be inspected under 365nm ultraviolet lamps, in test sample chromatography, on position corresponding with reference substance chromatography, the glimmering of same color is shown
Hot spot；
The thin-layer identification method of Herba Epimedii is：Drug granule content 5g is taken, 60-90 DEG C of petroleum ether 30ml is added, ultrasound
Processing 20 minutes, filtration, the dregs of a decoction wave most petroleum ether, add methanol 50ml, are heated to reflux 30 minutes, filter, and filter residue is washed with 5ml methanol
It washs, washing lotion merges with filtrate, is evaporated, and residue adds water 10ml, heating to make dissolving, let cool, set in centrifuge tube and centrifuge 10 minutes, take
Clear liquid, with chloroform shaking extraction 2 times, each 15ml discards chloroform liquid, shakes and extracts to water saturated n-butanol
3 times, add 10ml for the first time, 10ml, third time plus 5ml, merging n-butanol liquid is added to ammoniate test solution washing 3 times, every time for the second time
15ml discards ammoniacal liquor；N-butanol liquid is washed to neutrality to what n-butanol was saturated, discards aqueous, and n-butanol liquid is evaporated, and residue adds
Methanol 1ml makes dissolving, as test solution；Icariin reference substance is taken, adds methanol that solution of every 1ml containing 1mg is made, as
Reference substance solution；It is tested according to thin-layered chromatography (one VI B of annex of Chinese Pharmacopoeia version in 2010), draws 2~4 μ l of test solution
With 5 μ l of reference substance solution, put respectively on same silica gel g thin-layer plate, with chloroform-methanol-water=13:7:2,10 DEG C or less
Lower layer's solution of placement is solvent, is unfolded, and takes out, dries, and sprays with 10% ethanol solution of sulfuric acid, spot is heated at 105 DEG C
Colour developing is clear；In test sample chromatography, on position corresponding with reference substance chromatography, the spot of same color is shown；Setting wavelength again is
It is inspected under 365nm ultraviolet lamps, in test sample chromatography, on position corresponding with reference substance chromatography, shows the fluorescence of same color
Spot.
In the detection method of aforementioned Qijiaoshenbai capsule, the content assaying method of the matrine is：According to high-efficient liquid phase color
Spectrometry (four 0512 methods of Chinese Pharmacopoeia version in 2015) measures：
Chromatographic condition and system suitability：Using octadecylsilane chemically bonded silica as filler；Mobile phase：Methanol is
A phases, 0.1% triethylamine are B phases, elution requirement：53%A-47%B；Detection wavelength is 220nm；Theoretical cam curve presses matrine
Peak, which calculates, should be not less than 4000；
The preparation of reference substance solution：Precision weighs matrine reference substance, adds methanol that solution of every 1ml containing 0.2mg is made, shakes
It is even to get；
The preparation of test solution：Drug granule content 1.0g is taken, it is accurately weighed, it sets in conical flask with cover, is added dense
The strong ammonia solution 2ml wettings that degree is 25%-28%, are added chloroform 30ml, are heated to reflux 1.5 hours, filter, filtrate is steamed
Dry, residue adds 10ml methanol to dissolve, shake up to get；
Measuring method：Draw reference substance solution and each 5 μ l of test solution respectively, inject liquid chromatograph, measure to get.
In the detection method of aforementioned Qijiaoshenbai capsule, the content assaying method of the icariin is：According to efficient liquid phase
Chromatography (one VI D of annex of Chinese Pharmacopoeia version in 2010) measures：
Chromatographic condition and system suitability：Using octadecylsilane chemically bonded silica as filler；Mobile phase：Acetonitrile is
A phases, water are B phases, elution requirement：25.5%A-74.5%B；Detection wavelength is 270nm；Theoretical cam curve is based on icariin peak
3000 should be not less than by calculating；
The preparation of reference substance solution：Icariin reference substance is taken, it is accurately weighed, add methanol that every 1ml is made molten containing 25 μ g
Liquid to get；
The preparation of test solution：Drug substance contents are taken, after finely ground, take 1.5g, it is accurately weighed, it sets in conical flask with cover,
70% ethyl alcohol 25ml, close plug is added in precision, and weighed weight is ultrasonically treated, 1 hour, ultrasonic power 250W, supersonic frequency 35kHz,
Take out, let cool, then weighed weight, the weight of less loss supplied with 70% ethyl alcohol, is shaken up, filter, take subsequent filtrate to get；
Measuring method：It is accurate respectively to draw reference substance solution and each 10 μ l of test solution, liquid chromatograph is injected, is measured,
To obtain the final product.
In the detection method of aforementioned Qijiaoshenbai capsule, the drug detects in this way：
(1) character：Hard capsule content is the particle and powder of sepia；Bitter；
(2) differentiate：Include the thin layer of jujube is differentiated, the thin layer of Radix Angelicae Sinensis differentiates, the thin layer of kuh-seng differentiates, the thin layer of Radix Astragali
Differentiate and/or the thin layer of Herba Epimedii differentiates；
The thin layer of the jujube differentiates：Drug granule content 3g is taken, ethyl acetate 30ml is added, is ultrasonically treated 30 points
Clock, filtering, is evaporated filtrate, residue adds methanol 0.5ml to make dissolving, as test solution；Jujube control medicinal material 2g separately is taken, adds water
50ml is heated to reflux 2 hours, filters while hot, and filtrate volatilizes moisture and is made jujube thick paste, and cream adds ethyl acetate 30ml, at ultrasound
Reason 30 minutes, filtering, filtrate is evaporated, and adds methanol 1ml to make dissolving, as a contrast medicinal material solution, draws 5 μ l of test solution and right
According to 10 μ l of medicinal material solution, put respectively on same silica gel H lamellae, with toluene-ethyl acetate-glacial acetic acid=60:6:0.05 is
Solvent is unfolded, and takes out, dries, and sprays with 10% ethanol solution of sulfuric acid, is heated to that spot development is clear, and setting wavelength is at 105 DEG C
It is inspected under 365nm ultraviolet lamps, in test sample chromatography, on position corresponding with reference substance chromatography, shows the fluorescence of same color
Spot；
The thin-layer identification method of the Radix Angelicae Sinensis is：Drug granule content 5g is taken, n-hexane 20ml is added, is ultrasonically treated 20 points
Clock, filtration, filtrate is waved to 1ml, as test solution；Radix Angelicae Sinensis control medicinal material 0.5g separately is taken, is made in the same way of control medicinal material solution；
According to thin-layered chromatography (one VI B of annex of Chinese Pharmacopoeia version in 2010) test, draw each 5 μ l of above two solution, put respectively in
On same silica gel g thin-layer plate, with n-hexane-ethyl acetate=9:1 is solvent, is unfolded, and takes out, dries, and it is 365nm to set wavelength
It inspects under ultraviolet lamp, in test sample chromatography, is on the corresponding position of control medicinal material chromatography, the fluorescence spot of first same color；
The thin-layer identification method of the kuh-seng is：Drug granule content 1g is taken, ethyl alcohol 30ml is added, is heated to reflux 30 points
Clock is let cool, and filtration, filtrate is evaporated residue and water 20ml is added to make dissolving, is filtered, and filtrate is set in separatory funnel, and a concentration of 25%- is added
28% strong ammonia solution 0.5ml is merged chloroform liquid, is evaporated, residue adds with chloroform shaking extraction 2 times, each 10ml
Absolute ethyl alcohol 1ml makes dissolving, as test solution；Matrine and Sophoridine reference substance separately are taken, respectively plus absolute ethyl alcohol is made
Per solution of the 1ml containing 1mg, product solution, is tried according to thin-layered chromatography (one VI B of annex of Chinese Pharmacopoeia version in 2010) as a contrast
It tests, draws above-mentioned each 5 μ l of three kinds of solution, put respectively on same silica gel g thin-layer plate, it is dense with cyclohexane-acetone-ethyl acetate-
Ammonia solution=2:3:4:0.5 is solvent, is set in the pre-saturated exhibition cylinder of ammonia steam, presaturation 30 minutes, is unfolded, and takes out, dries,
Spray is with dilute bismuth potassium iodide test solution；In test sample chromatography, it is on the corresponding position of reference substance chromatography, shows the spot of same color；
The thin-layer identification method of the Radix Astragali is：Drug granule content 5g is taken, 60-90 DEG C of petroleum ether 30ml is added, is surpassed
Sonication 20 minutes, filtration, the dregs of a decoction wave most petroleum ether, add methanol 50ml, are heated to reflux 30 minutes, filter, a small amount of first of filter residue
Alcohol washs, and washing lotion merges with filtrate, is evaporated, and residue adds water 10ml, heating to make dissolving, let cool, set in centrifuge tube and centrifuge 10 minutes,
Supernatant is taken, with chloroform shaking extraction 2 times, each 15ml discards chloroform liquid, is shaken to water saturated n-butanol
Extraction 3 times adds 10ml for the first time, and 10ml, third time plus 5ml, merging n-butanol liquid is added to ammoniate test solution washing 3 times, often for the second time
Secondary 15ml, discards ammoniacal liquor；N-butanol liquid is washed to neutrality to what n-butanol was saturated, discards aqueous, and n-butanol liquid is evaporated, residue
Methanol 1ml is added to make dissolving, as test solution；Astragaloside IV reference substance separately is taken, adds methanol that solution of every 1ml containing 1mg is made,
Product solution as a contrast；It is tested according to thin-layered chromatography (one VI B of annex of Chinese Pharmacopoeia version in 2010), draws test solution 10
μ l, 5 μ l of reference substance solution are put respectively on same silica gel g thin-layer plate, with chloroform-methanol-water=13:7:2,10 DEG C with
Lower layer's solution of lower placement is solvent, is unfolded, and takes out, dries, and sprays with 10% ethanol solution of sulfuric acid, spot is heated at 105 DEG C
Point colour developing is clear；In test sample chromatography, on position corresponding with reference substance chromatography, the spot of same color is shown；Wavelength is set again
To be inspected under 365nm ultraviolet lamps, in test sample chromatography, on position corresponding with reference substance chromatography, the glimmering of same color is shown
Hot spot；
The thin-layer identification method of Herba Epimedii is：Drug granule content 5g is taken, 60-90 DEG C of petroleum ether 30ml is added, ultrasound
Processing 20 minutes, filtration, the dregs of a decoction wave most petroleum ether, add methanol 50ml, are heated to reflux 30 minutes, filter, and filter residue is washed with 5ml methanol
It washs, washing lotion merges with filtrate, is evaporated, and residue adds water 10ml, heating to make dissolving, let cool, set in centrifuge tube and centrifuge 10 minutes, take
Clear liquid, with chloroform shaking extraction 2 times, each 15ml discards chloroform liquid, shakes and extracts to water saturated n-butanol
3 times, add 10ml for the first time, 10ml, third time plus 5ml, merging n-butanol liquid is added to ammoniate test solution washing 3 times, every time for the second time
15ml discards ammoniacal liquor；N-butanol liquid is washed to neutrality to what n-butanol was saturated, discards aqueous, and n-butanol liquid is evaporated, and residue adds
Methanol 1ml makes dissolving, as test solution；Icariin reference substance is taken, adds methanol that solution of every 1ml containing 1mg is made, as
Reference substance solution；It is tested according to thin-layered chromatography (one VI B of annex of Chinese Pharmacopoeia version in 2010), draws 2~4 μ l of test solution
With 5 μ l of reference substance solution, put respectively on same silica gel g thin-layer plate, with chloroform-methanol-water=13:7:2,10 DEG C or less
Lower layer's solution of placement is solvent, is unfolded, and takes out, dries, and sprays with 10% ethanol solution of sulfuric acid, spot is heated at 105 DEG C
Colour developing is clear；In test sample chromatography, on position corresponding with reference substance chromatography, the spot of same color is shown；Setting wavelength again is
It is inspected under 365nm ultraviolet lamps, in test sample chromatography, on position corresponding with reference substance chromatography, shows the fluorescence of same color
Spot；
(3) assay：It include the assay to matrine and/or icariin；
The content assaying method of the matrine is：According to high performance liquid chromatography (Chinese Pharmacopoeia version in 2015 four 0512
Method) it measures：
Chromatographic condition and system suitability：Using octadecylsilane chemically bonded silica as filler；Mobile phase：Methanol is
A phases, 0.1% triethylamine are B phases, elution requirement：53%A-47%B；Detection wavelength is 220nm；Theoretical cam curve presses matrine
Peak, which calculates, should be not less than 4000；
The preparation of reference substance solution：Precision weighs matrine reference substance, adds methanol that solution of every 1ml containing 0.2mg is made, shakes
It is even to get；
The preparation of test solution：Drug granule content 1.0g is taken, it is accurately weighed, it sets in conical flask with cover, is added dense
The strong ammonia solution 2ml wettings that degree is 25%-28%, are added chloroform 30ml, are heated to reflux 1.5 hours, filter, filtrate is steamed
Dry, residue adds 10ml methanol to dissolve, shake up to get；
Measuring method：Draw reference substance solution and each 5 μ l of test solution respectively, inject liquid chromatograph, measure to get；
The content assaying method of the icariin is：According to high performance liquid chromatography, (Chinese Pharmacopoeia version one in 2010 is attached
Record VI D) it measures：
Chromatographic condition and system suitability：Using octadecylsilane chemically bonded silica as filler；Mobile phase：Acetonitrile is
A phases, water are B phases, elution requirement：25.5%A-74.5%B；Detection wavelength is 270nm；Theoretical cam curve is based on icariin peak
3000 should be not less than by calculating；
The preparation of reference substance solution：Icariin reference substance is taken, it is accurately weighed, add methanol that every 1ml is made molten containing 25 μ g
Liquid to get；
The preparation of test solution：The drug substance contents under content uniformity item are taken, it is finely ground, 1.5g is taken, it is accurately weighed, set tool
It fills in conical flask, 70% ethyl alcohol 25ml, close plug is added in precision, and weighed weight is ultrasonically treated, 1 hour, ultrasonic power 250W, is surpassed
Acoustic frequency 35kHz takes out, lets cool, then weighed weight, the weight of less loss is supplied with 70% ethyl alcohol, is shaken up, and filters, takes subsequent filtrate,
To obtain the final product；
Measuring method：It is accurate respectively to draw reference substance solution and each 10 μ l of test solution, liquid chromatograph is injected, is measured,
To obtain the final product.
Inventor has carried out a large amount of experiment, is the research of detection method of the present invention below
Experimental example 1：Jujube thin layer differentiates
1.1 instruments and reagent
ZF-20D uv analyzers；Silica G plate (100*100mm, lot number：20100321；100*200mm；Lot number：
20100317, producer：Haiyang Chemical Plant, Qingdao), silica gel H plate (100*100mm, lot number：20100503；100*200mm；Lot number：
20100417, producer：Haiyang Chemical Plant, Qingdao)；Silica gel H lamellae (self-control).
Absolute ethyl alcohol (lot number：20100705；Producer：Sinopharm Chemical Reagent Co., Ltd.), ethyl acetate (lot number：
1100101；Producer：Shanghai Bo Hua Co., Ltds), methanol (lot number：20100901；Producer：The limited public affairs of Chinese medicines group chemical reagent
Department), sulfuric acid (lot number：20090201；Producer：Chongqing Chuan Dong Chemical Co., Ltd.s), chloroform (lot number：0907142；Producer：
Xi Long science limited liability company), ether (lot number：20101125；Producer：Sinopharm Chemical Reagent Co., Ltd.), toluene
(lot number：20101225；Producer：Sinopharm Chemical Reagent Co., Ltd.), glacial acetic acid lot number：S189K040), ethyl acetate
(lot number：20090418；Chengdu Ke Long Chemical Co., Ltd.s).
Make lamellae by oneself：1 part of silica gel H is taken, is uniformly mixed with 1% 3 parts of CMC-Na aqueous solutions, bed board dries, 105
It DEG C dry 30 minutes, takes out, is put into drier, it is spare.
Qijiaoshenbai capsule (lot number：2527025,2527026,2528001), jujube negative sample (lot number：
20171218), jujube control medicinal material (lot number：121040-201408).Control medicinal material is purchased to Chinese pharmaceutical biological product and is identified
Institute；N-hexane, chloroform, methanol, ether, ethyl acetate, glacial acetic acid, toluene, ethyl alcohol etc. are that analysis is pure.
1.2 thin layer identification experiment conditions are investigated
1.2.1 test solution preparation condition is investigated
Condition 1：This product content 3g is taken, adds ethyl acetate 30ml ultrasounds 30 minutes, filtration, filtrate is evaporated, and residue adds first
Alcohol 0.5ml make dissolving to get.
Condition 2：This product content 3g is taken, the 30ml ultrasounds that add diethyl ether 30 minutes, filtration, filtrate is evaporated, and residue adds methanol
0.5ml make dissolving to get.
Condition 3：This product content 3g is taken, adds methanol 30ml ultrasounds 30 minutes, filtration, filtrate is evaporated, and residue adds methanol
0.5ml make dissolving to get.
1.2.2 prepared by control medicinal material solution
Condition 1：Jujube control medicinal material 2g is taken, ethyl acetate 30ml is added, is ultrasonically treated 30 minutes, filtering, filtrate is evaporated, residual
Slag add methanol 1ml dissolving to get.
Condition 2：Jujube control medicinal material 2g is taken, water 50ml is added, is heated to reflux 2 hours, filters while hot, filtrate volatilizes moisture system
At jujube thick paste, add ethyl acetate 50ml, be ultrasonically treated 30 minutes, filtering, filtrate is evaporated, add methanol 1ml make dissolving to get.
Condition 3：Jujube control medicinal material 2g is taken, water 50ml is added, is heated to reflux 2 hours, filters while hot, filtrate volatilizes moisture system
At jujube thick paste, add diethyl ether 50ml, is ultrasonically treated 30 minutes, and filtering, filtrate is evaporated, add methanol 1ml make dissolving to get.
Condition 4：Jujube control medicinal material 2g is taken, water 50ml is added, is heated to reflux 2 hours, filters while hot, filtrate volatilizes moisture system
At jujube thick paste, add methanol 50ml, be ultrasonically treated 30 minutes, filtering, filtrate is evaporated, add methanol 1ml make dissolving to get
1.2.3 prepared by negative test solution：The negative sample 3g for taking scarce jujube is prepared negative according to test solution preparation method
Contrast solution.
1.2.4 it is unfolded and inspects condition investigation
Condition 1：According to thin-layered chromatography (《Chinese Pharmacopoeia》Four 0502 methods of version in 2015[3]) experiment, control medicinal material is taken respectively
10 μ l of solution, 5 μ l points of test solution are on same silica gel H lamellae, respectively with toluene-ethyl acetate-glacial acetic acid (60:6:
0.05), n-hexane-ethyl acetate (3:1) it is solvent, is unfolded, takes out, dry, then spray with 10% sulfuric acid ethyl alcohol color developing agent,
105 DEG C are heated to clear spot, then set and inspected under ultraviolet lamp (365nm).
Condition 2：According to thin-layered chromatography (《Chinese Pharmacopoeia》Four 0502 methods of version in 2015) experiment, take control medicinal material molten respectively
10 μ l of liquid, 5 μ l points of test solution are on same silica gel g thin-layer plate, with toluene-ethyl acetate-glacial acetic acid (60:6:0.05) it is
Solvent is unfolded, and takes out, dries, and sprays with 10% sulfuric acid ethyl alcohol color developing agent, 105 DEG C are heated to clear spot, set ultraviolet lamp
It is inspected under (365nm).
Condition 3：According to thin-layered chromatography (《Chinese Pharmacopoeia》Four 0502 methods of version in 2015) experiment, the investigation of point sample amount：Respectively
Take 10 μ l of control medicinal material solution, 3 μ l of test solution, 5 μ l, 10 μ l, 15 μ l points on same silica gel H lamellae, with toluene-second
Acetoacetic ester-glacial acetic acid (60:6:0.05) it is solvent, is unfolded, takes out, dry, then spray with 10% sulfuric acid ethyl alcohol color developing agent,
105 DEG C are heated to clear spot, then set and inspected under ultraviolet lamp (365nm).
The result shows that taking this product content 3g, add ethyl acetate, ether, methanol that can propose jujube ingredient, from chromatogram
In as can be seen that ether, methanol are weaker than ethyl acetate, such as Fig. 1 to the extraction effect of jujube：
Compare from the chromatogram of chromatography condition and obtains, H lamellaes, toluene-ethyl acetate-glacial acetic acid (60:6:0.05)
Separating degree is good, and colour developing is clear, such as Fig. 2, Fig. 3, Fig. 4：5 μ l of point sample amount are that optimum condition investigates collection of illustrative plates such as Fig. 5：
In conjunction with above-mentioned experiment, this product content 3g is taken, adds ethyl acetate 30ml, is ultrasonically treated 30 minutes, filtering is evaporated filter
Liquid, residue adds methanol 0.5ml to make dissolving, as test sample.Jujube control medicinal material 2g separately is taken, adds water 50ml, it is small to be heated to reflux 2
When, it filters while hot, filtrate volatilizes moisture and jujube thick paste is made, and adds ethyl acetate 50ml, is ultrasonically treated 30 minutes, filtering, filtrate
It is evaporated, adds methanol 1ml to make dissolving, as a contrast medicinal material solution, draw 5 μ l of test solution, 10 μ l of control medicinal material solution, respectively
Point is on same silica gel H lamellae, with toluene-ethyl acetate-glacial acetic acid (60:6:0.05) it is solvent, is unfolded, takes out, dry in the air
Dry, with 10% ethanol solution of sulfuric acid, it is clear to be heated to spot development at 105 DEG C, sets and is inspected under ultraviolet lamp (365nm), is being supplied for spray
In test product chromatography, on position corresponding with reference substance chromatography, the fluorescence spot of same color is shown, spot separating degree is high, develops the color
Clearly, negative control is noiseless (such as Fig. 6), therefore selection is included in quality standard text.
1.3 durability
1.3.1 the comparison of different humidity
When 25 DEG C of temperature, it is unfolded under the conditions of humidity 35% and 75% respectively.As a result see (Fig. 7,8).
1.3.2 the comparison of different temperatures
When humidity 75%, it is unfolded under 4 DEG C, 10 DEG C and 40 DEG C of temperature environment respectively.As a result see (Fig. 9,10,
11)。
1.3.3 different label lamellaes (the result is shown in Figure 1 2- Figure 14)
2 determination of matrine of experimental example
2.1 instruments and reagent
Shimadzu LC-30AD high performance liquid chromatographs；1260 high performance liquid chromatograph of Agilent；KQ-500V DB type double frequency numbers
Control supersonic wave cleaning machine (Kunshan Ultrasonic Instruments Co., Ltd.)；Qijiaoshenbai capsule has Guizhou Hanfang Pharmaceutical Co., Ltd. to carry
For；Matrine reference substance (National Institute for Food and Drugs Control, lot number：110805-200508, for assay)；Reagent：First
Alcohol (chromatographically pure, analysis are pure), ethyl alcohol (analysis is pure), water (WP-UP-LH-40 physico-chemical analysis ultrapure water machine), triethylamine (analysis
It is pure), chloroform (analysis is pure), n-hexane (analysis is pure), ether (analysis is pure), phosphoric acid (analysis is pure).
2.2 assay method
1.2.1 chromatography condition：
(1) chromatographic column：With reference to Products Shu An wet tissues, Shu An washing lotions[5]Middle determination of matrine selection 18
Alkyl silane bonded silica gel is that the pillar of filler is investigated.The result shows that using octadecylsilane chemically bonded silica for filling
The pillar separating effect of agent is preferable.
(2) mobile phase：With reference to version pharmacopeia in 2015[4]And Products standard, grope through overtesting, using methanol-
0.1% triethylamine aqueous solution (53:47) it is mobile phase, ingredient matrine to be measured and other impurities are preferably detached.
(3) Detection wavelength：With reference to version pharmacopeia Sophora flavescens determination of matrine in 2015, select Detection wavelength for
220nm。
(4) theoretical cam curve：With reference to determination of matrine selection reason in Products Shu An wet tissues, Shu An washing lotions
It is no less than 4000 by the number of plates.
1.2.2 the preparation of test solution：
This product content 1.0g is taken, it is accurately weighed, it sets in conical flask with cover, strong ammonia solution 2ml wettings is added, trichlorine is added
Methane 30ml is heated to reflux 1.5 hours, and filtering, filtrate is evaporated, and residue adds 10ml methanol to dissolve, shake up to get.
1.2.3 the preparation of reference substance solution：
Precision weighs that matrine reference substance is appropriate, adds methanol that solution of every 1ml containing 0.2mg is made, shake up to get.
1.3 method validation
1.3.1 accuracy is tested
Take the Qijiaoshenbai capsule (lot number for having measured content：2527025, average content：1.7499mg/g) about 1.0g, essence
It is close weighed, it sets in conical flask with cover, matrine reference substance solution (0.22100mg/ml) 7ml is added in precision, by test solution
Preparation method prepares sample solution, and the content of matrine is measured by above-mentioned chromatographic condition, calculates the rate of recovery, the results are shown in Table 1 with
And accuracy chromatogram (Figure 15) sample size is 5 μ l, illustrates there is the preferable rate of recovery.
1 recovery test data (n=6) of table
1.3.2 repetitive test
It takes with a collection of Qijiaoshenbai capsule (2527025), 6 parts is prepared according to test solution preparation method, by above-mentioned color
Spectral condition measures the content of matrine, the results are shown in Table 2 and repeated chromatogram (Figure 16), illustrates there is preferable repeatability.
2 repeated experiment of table (n=6)
1.3.3 precision test
Precision is measured with a 5 μ l of Qijiaoshenbai capsule test solution, and continuous sample introduction 6 times records chromatogram, as a result sees
Table 3 and precision chromatogram (see Figure 17), illustrate there is preferable precision.
3 precision test of table (n=6)
1.3.4 specificity
Take Qijiaoshenbai capsule (2527025) and negative sample by test solution preparation method prepare sample solution with
And negative sample solution takes reference substance solution, sample solution, 5 μ l injection liquid of negative sample solution respectively by above-mentioned chromatographic condition
Chromatography, sample solution chromatography is in the corresponding chromatographic peak of reference substance solution chromatography, and feminine gender is noiseless, sees specificity chromatogram
(see Figure 18).
1.3.5 linear relationship is investigated
By above-mentioned chromatographic condition, precision take 3 μ l of matrine reference substance (0.22100mg/ml), 5 μ l, 7 μ l, 9 μ l, 11 μ l,
13 μ l are implanted sequentially liquid chromatograph, measure peak area.Using reference substance sample size X as abscissa, peak area Y is ordinate, into
Row linear regression calculates, and drawing curve (Figure 19) show that equation of linear regression is：Y=48.811X+17.542, R^2=
0.999.The matrine sample size of reference substance and peak area in the range of 0.663 μ g-2.873 μ g are in good linear relationship.
See linear chromatography figure (Figure 20).
4 matrine reference substance linear relationship of table
1.3.6 range
By normal 80% and 120% sampling and measuring for measuring sampling amount, the precision being measured by sampling in height is investigated.Remove stilbene
Glue paste capsule of rising white blood cell (2527025) is just sampling each 6 parts, is prepared by test solution preparation method, is surveyed by above-mentioned chromatographic condition
The content for determining matrine, the results are shown in Table 5.
5 range finding result of table
1.3.7 durability
1.3.7.1 the selection of sample extraction solvent
Qijiaoshenbai capsule (2527025) sample is taken, it is finely ground, it is uniformly mixed, takes 3 parts, every part of about 1.0g, it is accurately weighed,
It sets in conical flask with cover respectively, after being separately added into the wetting of 2ml strong ammonia solutions, then accurate addition ether, chloroform, methanol are each
30ml is heated to reflux 1.5 hours, is taken out, and filtering, filtrate is evaporated, and residue adds methanol 10ml to dissolve, and obtains test solution, respectively
Each 5 μ l injections liquid chromatograph of above-mentioned 4 parts of solution is taken, calculates, the results are shown in Table 6, illustrate this product using chloroform as extraction
Solvent, content extraction are more complete.
The selection of 6 Extraction solvent of table
1.3.7.2 the selection of sample extraction method
Qijiaoshenbai capsule (2527025) sample is taken, it is finely ground, it is uniformly mixed, takes 2 parts every part about 1.0g, it is accurately weighed, point
Do not set in conical flask with cover, be separately added into it is accurate after the wetting of 2ml strong ammonia solutions be added chloroform 30ml, at a copy of it ultrasound
Reason 1 hour, another heating and refluxing extraction 1 hour are taken out, and filtering, filtrate is evaporated.Residue adds methanol 10ml to dissolve, and obtains for examination
Product solution takes above-mentioned 2 parts of solution each 5 μ l injection liquid chromatograph, calculates, the results are shown in Table 7 respectively, is ultrasonically treated and refluxing extraction
Comparision contents, refluxing extraction content is more complete, therefore selects heating and refluxing extraction.
The selection of 7 extracting method of table
1.3.7.3 sample extraction time effects
Qijiaoshenbai capsule (2527025) sample is taken, it is finely ground, it is uniformly mixed, takes 3 parts, every part of about 1.0g is accurately weighed,
It sets in conical flask with cover respectively, respectively after the wetting of 2ml strong ammonia solutions, then accurate addition chloroform 30ml, it is heated to reflux respectively
1.0, it 1.5,2.0 hours, takes out, filtering, filtrate is evaporated, and residue adds methanol 10ml to dissolve, and obtains test solution, takes respectively above-mentioned
Each 5 μ l of 3 parts of solution inject liquid chromatograph, calculate, the results are shown in Table 8, consider, select to be heated to reflux 1.5 hours as sample
Extraction time.
8 extraction time of table influences
1.3.7.4 the influence of different brands pillar
Different brands octadecylsilane chemically bonded silica is used to determine Qijiaoshenbai capsule respectively for the pillar of filler
(2527025) content of matrine in, investigates the influence of the chromatographic column of different brands.Chromatogram is shown in durability chromatogram, content
Measurement result is shown in Table 9.The result shows that different brands octadecylsilane chemically bonded silica be filler chromatographic column to assay
Without influence.
The measurement result of the pillar of the different labels of table 9
1.3.7.5 the influence of mobile phase ratio
In the case where not changing mobile phase member condition, change mobile phase ratio, investigates the influence of different proportion mobile phase, as a result
Under selected mobile phase ratio, sample separating degree is preferable, and assay the results are shown in Table 10 and Figure 21.
Measurement result under the conditions of the different mobile phase ratios of table 10
1.3.7.6 sample stability is tested
Qijiaoshenbai capsule (2527025) is taken, is prepared by test solution preparation method, is measured by above-mentioned chromatographic condition bitter
The content for joining alkali, the results are shown in Table 11 and Figure 22.
11 sample stability of table is tested
1.4 samples measure
By above-mentioned condition, 15 batches of samples are measured respectively, the results are shown in Table 12.
12 sample measurement result of table
This product Sophora flavescens are used as medicine for extract, therefore per 1g containing kuh-seng with matrine (C15H24N2O it) counts, content limit is answered
For：Kuh-seng content * Sophora flavescens limit (matrine, Oxymatrine total alkali content)/obtained capsule grain number * 0.7=in prescription
250g*1.2%/1000 * 0.7*1000mg=2.10mg/, every 0.5g, therefore calculated by per 1g, it is 4.2mg/g.
Due to kuh-seng boiling in the production process, matrine has partial loss and cream powder to mix during being somebody's turn to do, and 75
Kuh-seng alkali loss is serious in~95 DEG C of drying processes, and being tested in conjunction with production process proves, boiling matrine transfer during two
Up to 58%, total rate of transform mixed matrine in drying process and can only achieve 41.5%, total rate of transform can only fail rate of about 40%
The rate of transform for reaching 70% is shown in Table 13 simulation matrine Transfer Experiment results and matrine Transfer Experiment chromatogram (Figure 23).
Therefore tentative this product is per 1g (C containing matrine15H24N2O it) counts, 1.3mg/g must not be less than.
Table 13 simulates matrine Transfer Experiment result
The beneficial effects of the present invention are：The present invention provides a kind of detection methods of Qijiaoshenbai capsule, including to have
It is index to imitate ingredient matrine and icariin, using the content of high effective liquid chromatography for measuring matrine and icariin；With
And in drug jujube, Radix Angelicae Sinensis, kuh-seng, Radix Astragali, Herba Epimedii thin-layer chromatography (TLC) discrimination method；The detection method is accurate, spirit
Sensitivity is high, reproducible, and testing result is stablized, and increases jujube indentification by TLC；Measure Sophora flavescens in matrine amount, be
Effectively reliable quality standard is established, Chinese medicine total quality can be accurately reflected, data reference is provided.Stilbene glue liter can effectively be controlled
The quality of white glue capsule had both been more advantageous to the monitoring of manufacturer and supervisory and management department to product quality, or medical portion
The treatment of door and patient, which provide, preferably to be ensured.
Description of the drawings
Fig. 1 be expansion and inspect condition investigate result figure (1, jujube extraction conditions 2；2 jujube extraction conditions 3；Jujube extracts
Condition；4, Qijiaoshenbai capsule extraction conditions 1；5, Qijiaoshenbai capsule extraction conditions 2；6, Qijiaoshenbai capsule extraction conditions
3)；
Fig. 2 is H plate toluene：Ethyl acetate：Glacial acetic acid=60:6:When 0.05, from the chromatogram of chromatography condition；(1, jujube
Control medicinal material solution；2,2527025；3,2527026；4,2528001)；
Fig. 3 is H plate n-hexanes：Ethyl acetate (3:1) when, from the chromatogram of chromatography condition；(1, jujube control medicinal material it is molten
Liquid；2,2527025；3,2527026；4,2528001)；
Fig. 4 is G lamellaes, toluene：Ethyl acetate：Glacial acetic acid (60:6:0.05) when, from the chromatogram of chromatography condition；(1,
Jujube control medicinal material solution；2,2527025；3,2527026；4,2528001)；
Fig. 5 be different point sample spirograms spectrum (1,10 μ l of jujube control medicinal material solution；2,3 μ l of test sample；3,5 μ l of test sample；4,
10 μ l of test sample；5,15 μ l of test sample)；
Fig. 6 be specificity collection of illustrative plates (1, jujube control medicinal material solution 2,25270253,25270264,2,528,001 5, lack it is big
Jujube negative sample)；
When Fig. 7 is 25 DEG C of temperature, humidity 35%, durability result figure (1, jujube control medicinal material solution；2,2527025；3,
2527026；4,2528001)；
When Fig. 8 is 25 DEG C of temperature, humidity 75%, durability result figure (1, jujube control medicinal material solution；2,2527025；3,
2527026；4,2528001)；
When Fig. 9 is humidity 75%, 4 DEG C of temperature, durability result figure (1, jujube control medicinal material solution；2,2527025；3,
2527026；4,2528001)；
When Figure 10 is humidity 75%, 10 DEG C of temperature, durability result figure (1, jujube control medicinal material solution；2,2527025；
3,2527026；4,2528001)；
When Figure 11 is humidity 75%, 40 DEG C of temperature, durability result figure (1, jujube control medicinal material solution；2,2527025；
3,2527026；4,2528001)；
When Figure 12 is humidity 75%, 25 DEG C of temperature, Haiyang Chemical Plant, Qingdao's board lamellae durability result figure (1, jujube pair
According to medicinal material solution；2,2527025；3,2527026；4,2528001)；
When Figure 13 is humidity 75%, 25 DEG C of temperature, Qingdao wave chemical industry label lamellae durability result figure；(1, jujube
Control medicinal material solution 2,2,527,025 3,2,527,026 4,2528001)
When Figure 14 is humidity 75%, 25 DEG C of temperature, self-control H lamellae durabilities result figure (1, jujube control medicinal material solution
2,2527025；2527026 4,2528001)
Figure 15 is matrine accuracy chromatogram；(cis-matrine control 1, test number (TN) 1；B- matrines control 2, experiment
Number 2；C- matrines control 3, test number (TN) 3；D-Matrine control 4, test number (TN) 4；E- matrines control 5, test number (TN)
5；F- matrines control 6, test number (TN) 6)；
Figure 16 is matrine repeatability chromatogram (cis-matrine control 1, test number (TN) 1；B- matrines control 2, experiment time
Number 2；C- matrines control 3, test number (TN) 3；D-Matrine control 4, test number (TN) 4；E- matrines control 5, test number (TN) 5；
F- matrines control 6, test number (TN) 6)；
Figure 17 is matrine precision chromatogram (cis-matrine control 1, test number (TN) 1；B- matrines control 2, experiment time
Number 2；C- matrines control 3, test number (TN) 3；D-Matrine control 4, test number (TN) 4；E- matrines control 5, test number (TN) 5；
F- matrines control 6, test number (TN) 6)；
Figure 18 is matrine specificity chromatogram (cis-matrine control 1, test number (TN) 1；B- matrines control 2, experiment time
Number 2；3) c- matrines compare；
When Figure 19 is determination of matrine, linear regression curves figure；
When Figure 20 is determination of matrine, linear chromatography figure (cis-matrine control 1, test number (TN) 1；B- matrines pair
According to 2, test number (TN) 2；C- matrines control 3, test number (TN) 3；D-Matrine control 4, test number (TN) 4；E- matrines control 5,
Test number (TN) 5；F- matrines control 6, test number (TN) 6)；
When Figure 21 is determination of matrine, mobile phase ratio influences result figure；(a- methanol：0.1% triethylamine aqueous solution
=53：47；B- methanol：0.1% triethylamine aqueous solution=50:45；C- methanol：0.1% triethylamine aqueous solution=50:50)；
When Figure 22 is determination of matrine, stability chromatogram；
When Figure 23 is determination of matrine, matrine processing simulation chromatogram；Wherein a is 1 (kuh-seng of processing simulation sequence
Alkali reference substance)；B is processing simulation sequence 2 (25278001 total mixed preceding medicinal extract)；C is that (25278001 is total mixed for processing simulation sequence 3
Afterwards)；D is processing simulation sequence 4 (25278002 total mixed preceding medicinal extract)；E is processing simulation sequence 5 (25278002 is total mixed rear)；F is
Processing simulation sequence 6 (matrine, oxymatrine mixing control)；G is 7 (Sophora flavescens of processing simulation sequence
ZY17112001)；H is processing simulation sequence 8 (Sophora flavescens ZY17112002).
With reference to embodiment, the present invention is further illustrated
Specific implementation mode
Embodiment 1：The detection method of Qijiaoshenbai capsule is
Formula：Jujube 250g, donkey-hide gelatin 250g, buerger lespedeza root 375g, Herba Epimedii 375g, kuh-seng 250g, Radix Astragali 750g, Radix Angelicae Sinensis
250g。
Technique：Above seven taste, in addition to donkey-hide gelatin, Radix Angelicae Sinensis is ground into fine powder；The five tastes such as remaining jujube add water to cook three times, the
One time 2 hours, second 1.5 hours, third time 1 hour, collecting decoction filtered, and the donkey-hide gelatin after molten is added in filtrate, stirs evenly,
It is concentrated into the thick paste that relative density is 1.30 at 75-85 DEG C, above-mentioned Radix Angelicae Sinensis fine powder is added, particle is made in mixing, dry, is packed into
Capsule to get.
Detection method：
(1) character：Hard capsule content is the particle and powder of sepia；Bitter；
(2) differentiate：Include the thin layer of jujube is differentiated, the thin layer of Radix Angelicae Sinensis differentiates, the thin layer of kuh-seng differentiates, the thin layer of Radix Astragali
Differentiate and/or the thin layer of Herba Epimedii differentiates；
The thin layer of the jujube differentiates：Drug granule content 3g is taken, ethyl acetate 30ml is added, is ultrasonically treated 30 points
Clock, filtering, is evaporated filtrate, residue adds methanol 0.5ml to make dissolving, as test solution；Jujube control medicinal material 2g separately is taken, adds water
50ml is heated to reflux 2 hours, filters while hot, and filtrate volatilizes moisture and is made jujube thick paste, and cream adds ethyl acetate 30ml, at ultrasound
Reason 30 minutes, filtering, filtrate is evaporated, and adds methanol 1ml to make dissolving, as a contrast medicinal material solution, draws 5 μ l of test solution and right
According to 10 μ l of medicinal material solution, put respectively on same silica gel H lamellae, with toluene-ethyl acetate-glacial acetic acid=60:6:0.05 is
Solvent is unfolded, and takes out, dries, and sprays with 10% ethanol solution of sulfuric acid, is heated to that spot development is clear, and setting wavelength is at 105 DEG C
It is inspected under 365nm ultraviolet lamps, in test sample chromatography, on position corresponding with reference substance chromatography, shows the fluorescence of same color
Spot；
The thin-layer identification method of the Radix Angelicae Sinensis is：Drug granule content 5g is taken, n-hexane 20ml is added, is ultrasonically treated 20 points
Clock, filtration, filtrate is waved to 1ml, as test solution；Radix Angelicae Sinensis control medicinal material 0.5g separately is taken, is made in the same way of control medicinal material solution；
According to thin-layered chromatography (one VI B of annex of Chinese Pharmacopoeia version in 2010) test, draw each 5 μ l of above two solution, put respectively in
On same silica gel g thin-layer plate, with n-hexane-ethyl acetate=9:1 is solvent, is unfolded, and takes out, dries, and it is 365nm to set wavelength
It inspects under ultraviolet lamp, in test sample chromatography, is on the corresponding position of control medicinal material chromatography, the fluorescence spot of first same color；
The thin-layer identification method of the kuh-seng is：Drug granule content 1g is taken, ethyl alcohol 30ml is added, is heated to reflux 30 points
Clock is let cool, and filtration, filtrate is evaporated residue and water 20ml is added to make dissolving, is filtered, and filtrate is set in separatory funnel, and a concentration of 25%- is added
28% strong ammonia solution 0.5ml is merged chloroform liquid, is evaporated, residue adds with chloroform shaking extraction 2 times, each 10ml
Absolute ethyl alcohol 1ml makes dissolving, as test solution；Matrine and Sophoridine reference substance separately are taken, respectively plus absolute ethyl alcohol is made
Per solution of the 1ml containing 1mg, product solution, is tried according to thin-layered chromatography (one VI B of annex of Chinese Pharmacopoeia version in 2010) as a contrast
It tests, draws above-mentioned each 5 μ l of three kinds of solution, put respectively on same silica gel g thin-layer plate, it is dense with cyclohexane-acetone-ethyl acetate-
Ammonia solution=2:3:4:0.5 is solvent, is set in the pre-saturated exhibition cylinder of ammonia steam, presaturation 30 minutes, is unfolded, and takes out, dries,
Spray is with dilute bismuth potassium iodide test solution；In test sample chromatography, it is on the corresponding position of reference substance chromatography, shows the spot of same color；
The thin-layer identification method of the Radix Astragali is：Drug granule content 5g is taken, 60-90 DEG C of petroleum ether 30ml is added, is surpassed
Sonication 20 minutes, filtration, the dregs of a decoction wave most petroleum ether, add methanol 50ml, are heated to reflux 30 minutes, filter, a small amount of first of filter residue
Alcohol washs, and washing lotion merges with filtrate, is evaporated, and residue adds water 10ml, heating to make dissolving, let cool, set in centrifuge tube and centrifuge 10 minutes,
Supernatant is taken, with chloroform shaking extraction 2 times, each 15ml discards chloroform liquid, is shaken to water saturated n-butanol
Extraction 3 times adds 10ml for the first time, and 10ml, third time plus 5ml, merging n-butanol liquid is added to ammoniate test solution washing 3 times, often for the second time
Secondary 15ml, discards ammoniacal liquor；N-butanol liquid is washed to neutrality to what n-butanol was saturated, discards aqueous, and n-butanol liquid is evaporated, residue
Methanol 1ml is added to make dissolving, as test solution；Astragaloside IV reference substance separately is taken, adds methanol that solution of every 1ml containing 1mg is made,
Product solution as a contrast；It is tested according to thin-layered chromatography (one VI B of annex of Chinese Pharmacopoeia version in 2010), draws test solution 10
μ l, 5 μ l of reference substance solution are put respectively on same silica gel g thin-layer plate, with chloroform-methanol-water=13:7:2,10 DEG C with
Lower layer's solution of lower placement is solvent, is unfolded, and takes out, dries, and sprays with 10% ethanol solution of sulfuric acid, spot is heated at 105 DEG C
Point colour developing is clear；In test sample chromatography, on position corresponding with reference substance chromatography, the spot of same color is shown；Wavelength is set again
To be inspected under 365nm ultraviolet lamps, in test sample chromatography, on position corresponding with reference substance chromatography, the glimmering of same color is shown
Hot spot；
The thin-layer identification method of Herba Epimedii is：Drug granule content 5g is taken, 60-90 DEG C of petroleum ether 30ml is added, ultrasound
Processing 20 minutes, filtration, the dregs of a decoction wave most petroleum ether, add methanol 50ml, are heated to reflux 30 minutes, filter, and filter residue is washed with 5ml methanol
It washs, washing lotion merges with filtrate, is evaporated, and residue adds water 10ml, heating to make dissolving, let cool, set in centrifuge tube and centrifuge 10 minutes, take
Clear liquid, with chloroform shaking extraction 2 times, each 15ml discards chloroform liquid, shakes and extracts to water saturated n-butanol
3 times, add 10ml for the first time, 10ml, third time plus 5ml, merging n-butanol liquid is added to ammoniate test solution washing 3 times, every time for the second time
15ml discards ammoniacal liquor；N-butanol liquid is washed to neutrality to what n-butanol was saturated, discards aqueous, and n-butanol liquid is evaporated, and residue adds
Methanol 1ml makes dissolving, as test solution；Icariin reference substance is taken, adds methanol that solution of every 1ml containing 1mg is made, as
Reference substance solution；It is tested according to thin-layered chromatography (one VI B of annex of Chinese Pharmacopoeia version in 2010), draws 2~4 μ l of test solution
With 5 μ l of reference substance solution, put respectively on same silica gel g thin-layer plate, with chloroform-methanol-water=13:7:2,10 DEG C or less
Lower layer's solution of placement is solvent, is unfolded, and takes out, dries, and sprays with 10% ethanol solution of sulfuric acid, spot is heated at 105 DEG C
Colour developing is clear；In test sample chromatography, on position corresponding with reference substance chromatography, the spot of same color is shown；Setting wavelength again is
It is inspected under 365nm ultraviolet lamps, in test sample chromatography, on position corresponding with reference substance chromatography, shows the fluorescence of same color
Spot；
(3) assay：It include the assay to matrine and/or icariin；
The content assaying method of the matrine is：According to high performance liquid chromatography (Chinese Pharmacopoeia version in 2015 four 0512
Method) it measures：
Chromatographic condition and system suitability：Using octadecylsilane chemically bonded silica as filler；Mobile phase：Methanol is
A phases, 0.1% triethylamine are B phases, elution requirement：53%A-47%B；Detection wavelength is 220nm；Theoretical cam curve presses matrine
Peak, which calculates, should be not less than 4000；
The preparation of reference substance solution：Precision weighs matrine reference substance, adds methanol that solution of every 1ml containing 0.2mg is made, shakes
It is even to get；
The preparation of test solution：Drug granule content 1.0g is taken, it is accurately weighed, it sets in conical flask with cover, is added dense
The strong ammonia solution 2ml wettings that degree is 25%-28%, are added chloroform 30ml, are heated to reflux 1.5 hours, filter, filtrate is steamed
Dry, residue adds 10ml methanol to dissolve, shake up to get；
Measuring method：Draw reference substance solution and each 5 μ l of test solution respectively, inject liquid chromatograph, measure to get；
The content assaying method of the icariin is：According to high performance liquid chromatography, (Chinese Pharmacopoeia version one in 2010 is attached
Record VI D) it measures：
Chromatographic condition and system suitability：Using octadecylsilane chemically bonded silica as filler；Mobile phase：Acetonitrile is
A phases, water are B phases, elution requirement：25.5%A-74.5%B；Detection wavelength is 270nm；Theoretical cam curve is based on icariin peak
3000 should be not less than by calculating；
The preparation of reference substance solution：Icariin reference substance is taken, it is accurately weighed, add methanol that every 1ml is made molten containing 25 μ g
Liquid to get；
The preparation of test solution：The drug substance contents under content uniformity item are taken, it is finely ground, 1.5g is taken, it is accurately weighed, set tool
It fills in conical flask, 70% ethyl alcohol 25ml, close plug is added in precision, and weighed weight is ultrasonically treated, 1 hour, ultrasonic power 250W, is surpassed
Acoustic frequency 35kHz takes out, lets cool, then weighed weight, the weight of less loss is supplied with 70% ethyl alcohol, is shaken up, and filters, takes subsequent filtrate,
To obtain the final product；
Measuring method：It is accurate respectively to draw reference substance solution and each 10 μ l of test solution, liquid chromatograph is injected, is measured,
To obtain the final product.
Claims (6)
1. a kind of detection method of Qijiaoshenbai capsule, it is characterised in that：The drug is by 240-260 parts of jujube, donkey-hide gelatin 240-
250 parts, 365-385 parts of buerger lespedeza root, 365-385 parts of Herba Epimedii, 240-260 parts of kuh-seng, 740-760 parts of Radix Astragali and Radix Angelicae Sinensis 240-
260 parts are prepared by the following method：Above seven taste, in addition to donkey-hide gelatin, Radix Angelicae Sinensis is ground into fine powder；The five tastes such as remaining jujube, add
Water decocts three times, 2 hours for the first time, second 1.5 hours, third time 1 hour, collecting decoction, filtration, after molten is added in filtrate
Donkey-hide gelatin, stir evenly, be concentrated into the thick paste that relative density at 75-85 DEG C is 1.30, be added above-mentioned Radix Angelicae Sinensis fine powder, mixing is made
Grain, it is dry, be packed into capsule to get；The detection method includes discrimination method and content assaying method；The discrimination method includes
To the thin of the thin layer discriminating of jujube, the thin layer discriminating of Radix Angelicae Sinensis, the thin layer discriminating of kuh-seng, the thin layer discriminating of Radix Astragali and/or Herba Epimedii
Layer differentiates；The content assaying method includes the assay to matrine and/or icariin.
2. the detection method of Qijiaoshenbai capsule as described in claim 1, it is characterised in that：The thin layer of the jujube differentiates：
Drug granule content 3g is taken, ethyl acetate 30ml is added, is ultrasonically treated 30 minutes, filtering is evaporated filtrate, residue adds methanol
0.5ml makes dissolving, as test solution；Jujube control medicinal material 2g separately is taken, adds water 50ml, is heated to reflux 2 hours, while hot mistake
Filter, filtrate volatilize moisture and jujube thick paste are made, and cream adds ethyl acetate 30ml, is ultrasonically treated 30 minutes, and filtering, filtrate is evaporated, adds
Methanol 1ml makes dissolving, as a contrast medicinal material solution, draws 5 μ l of test solution and 10 μ l of control medicinal material solution, puts respectively in same
On one silica gel H lamellae, with toluene-ethyl acetate-glacial acetic acid=60:6:0.05 is solvent, be unfolded, take out, dry, spray with
10% ethanol solution of sulfuric acid, it is clear to be heated to spot development at 105 DEG C, and it is to be inspected under 365nm ultraviolet lamps to set wavelength, in test sample
In chromatography, on position corresponding with reference substance chromatography, the fluorescence spot of same color is shown.
3. the detection method of Qijiaoshenbai capsule as described in claim 1, it is characterised in that：The thin-layer identification method of the Radix Angelicae Sinensis
For：Drug granule content 5g is taken, n-hexane 20ml is added, is ultrasonically treated 20 minutes, filtration, filtrate is waved to 1ml, as test sample
Solution；Radix Angelicae Sinensis control medicinal material 0.5g separately is taken, is made in the same way of control medicinal material solution；According to thin-layered chromatography (Chinese Pharmacopoeia version in 2010
One VI B of annex) experiment, each 5 μ l of above two solution are drawn, are put respectively on same silica gel g thin-layer plate, with n-hexane-second
Acetoacetic ester=9:1 is solvent, is unfolded, and takes out, dries, and it is to be inspected under 365nm ultraviolet lamps to set wavelength, in test sample chromatography,
In on the corresponding position of control medicinal material chromatography, the fluorescence spot of first same color；
The thin-layer identification method of the kuh-seng is：Drug granule content 1g is taken, ethyl alcohol 30ml is added, is heated to reflux 30 minutes, puts
Cold, filtration, filtrate is evaporated residue and water 20ml is added to make dissolving, filters, and filtrate is set in separatory funnel, adds a concentration of 25%-28%'s
Strong ammonia solution 0.5ml is merged chloroform liquid, is evaporated, residue adds anhydrous second with chloroform shaking extraction 2 times, each 10ml
Alcohol 1ml makes dissolving, as test solution；Matrine and Sophoridine reference substance separately are taken, respectively plus absolute ethyl alcohol is made every 1ml and contains
The solution of 1mg, product solution, is tested according to thin-layered chromatography (one VI B of annex of Chinese Pharmacopoeia version in 2010) as a contrast, in absorption
Each 5 μ l of three kinds of solution are stated, are put respectively on same silica gel g thin-layer plate, with cyclohexane-acetone-ethyl acetate-strong ammonia solution=2:
3:4:0.5 is solvent, is set in the pre-saturated exhibition cylinder of ammonia steam, presaturation 30 minutes, is unfolded, and takes out, dries, and is sprayed with dilute iodate
Bismuth potassium test solution；In test sample chromatography, it is on the corresponding position of reference substance chromatography, shows the spot of same color；
The thin-layer identification method of the Radix Astragali is：Drug granule content 5g is taken, adds 60-90 DEG C of petroleum ether 30ml, at ultrasound
Reason 20 minutes, filtration, the dregs of a decoction wave most petroleum ether, add methanol 50ml, are heated to reflux 30 minutes, filter, and filter residue is washed with a small amount of methanol
It washs, washing lotion merges with filtrate, is evaporated, and residue adds water 10ml, heating to make dissolving, let cool, set in centrifuge tube and centrifuge 10 minutes, take
Clear liquid, with chloroform shaking extraction 2 times, each 15ml discards chloroform liquid, shakes and extracts to water saturated n-butanol
3 times, add 10ml for the first time, 10ml, third time plus 5ml, merging n-butanol liquid is added to ammoniate test solution washing 3 times, every time for the second time
15ml discards ammoniacal liquor；N-butanol liquid is washed to neutrality to what n-butanol was saturated, discards aqueous, and n-butanol liquid is evaporated, and residue adds
Methanol 1ml makes dissolving, as test solution；Astragaloside IV reference substance separately is taken, adds methanol that solution of every 1ml containing 1mg is made, is made
For reference substance solution；It is tested according to thin-layered chromatography (one VI B of annex of Chinese Pharmacopoeia version in 2010), draws 10 μ of test solution
L, 5 μ l of reference substance solution are put respectively on same silica gel g thin-layer plate, with chloroform-methanol-water=13:7:2,10 DEG C or less
Lower layer's solution of placement is solvent, is unfolded, and takes out, dries, and sprays with 10% ethanol solution of sulfuric acid, spot is heated at 105 DEG C
Colour developing is clear；In test sample chromatography, on position corresponding with reference substance chromatography, the spot of same color is shown；Setting wavelength again is
It is inspected under 365nm ultraviolet lamps, in test sample chromatography, on position corresponding with reference substance chromatography, shows the fluorescence of same color
Spot；
The thin-layer identification method of Herba Epimedii is：Drug granule content 5g is taken, 60-90 DEG C of petroleum ether 30ml is added, is ultrasonically treated
20 minutes, filtration, the dregs of a decoction waved most petroleum ether, add methanol 50ml, are heated to reflux 30 minutes, filtered, and filter residue is washed with 5ml methanol,
Washing lotion merges with filtrate, is evaporated, and residue adds water 10ml, heating to make dissolving, let cool, set in centrifuge tube and centrifuge 10 minutes, take supernatant
Liquid, with chloroform shaking extraction 2 times, each 15ml discards chloroform liquid, to water saturated n-butanol shaking extraction 3
It is secondary, add 10ml for the first time, 10ml, third time plus 5ml, merging n-butanol liquid is added to ammoniate test solution washing 3 times, every time for the second time
15ml discards ammoniacal liquor；N-butanol liquid is washed to neutrality to what n-butanol was saturated, discards aqueous, and n-butanol liquid is evaporated, and residue adds
Methanol 1ml makes dissolving, as test solution；Icariin reference substance is taken, adds methanol that solution of every 1ml containing 1mg is made, as
Reference substance solution；It is tested according to thin-layered chromatography (one VI B of annex of Chinese Pharmacopoeia version in 2010), draws 2~4 μ l of test solution
With 5 μ l of reference substance solution, put respectively on same silica gel g thin-layer plate, with chloroform-methanol-water=13:7:2,10 DEG C or less
Lower layer's solution of placement is solvent, is unfolded, and takes out, dries, and sprays with 10% ethanol solution of sulfuric acid, spot is heated at 105 DEG C
Colour developing is clear；In test sample chromatography, on position corresponding with reference substance chromatography, the spot of same color is shown；Setting wavelength again is
It is inspected under 365nm ultraviolet lamps, in test sample chromatography, on position corresponding with reference substance chromatography, shows the fluorescence of same color
Spot.
4. the detection method of Qijiaoshenbai capsule as described in claim 1, it is characterised in that：The assay side of the matrine
Method is：It is measured according to high performance liquid chromatography (four 0512 methods of Chinese Pharmacopoeia version in 2015)：
Chromatographic condition and system suitability：Using octadecylsilane chemically bonded silica as filler；Mobile phase：Methanol is A phases,
0.1% triethylamine is B phases, elution requirement：53%A-47%B；Detection wavelength is 220nm；Theoretical cam curve is based on matrine peak
4000 should be not less than by calculating；
The preparation of reference substance solution：Precision weighs matrine reference substance, adds methanol that solution of every 1ml containing 0.2mg is made, shakes up,
To obtain the final product；
The preparation of test solution：Drug granule content 1.0g is taken, it is accurately weighed, it sets in conical flask with cover, is added a concentration of
The strong ammonia solution 2ml wettings of 25%-28%, are added chloroform 30ml, are heated to reflux 1.5 hours, filter, filtrate is evaporated, residual
Slag add 10ml methanol dissolve, shake up to get；
Measuring method：Draw reference substance solution and each 5 μ l of test solution respectively, inject liquid chromatograph, measure to get.
5. the detection method as described in claim 1 for stating Qijiaoshenbai capsule, it is characterised in that：The content of the icariin
Assay method is：It is measured according to high performance liquid chromatography (one VI D of annex of Chinese Pharmacopoeia version in 2010)：
Chromatographic condition and system suitability：Using octadecylsilane chemically bonded silica as filler；Mobile phase：Acetonitrile is A phases,
Water is B phases, elution requirement：25.5%A-74.5%B；Detection wavelength is 270nm；Theoretical cam curve is answered by the calculating of icariin peak
Not less than 3000；
The preparation of reference substance solution：Icariin reference substance is taken, it is accurately weighed, add methanol that the solution that every 1ml contains 25 μ g is made, i.e.,
?；
The preparation of test solution：Drug substance contents are taken, after finely ground, take 1.5g, it is accurately weighed, it sets in conical flask with cover, it is accurate
70% ethyl alcohol 25ml, close plug is added, weighed weight is ultrasonically treated, takes within 1 hour, ultrasonic power 250W, supersonic frequency 35kHz
Go out, let cool, then weighed weight, the weight of less loss is supplied with 70% ethyl alcohol, is shaken up, filter, take subsequent filtrate to get；
Measuring method：It is accurate respectively to draw reference substance solution and each 10 μ l of test solution, inject liquid chromatograph, measure to get.
6. the detection method of Qijiaoshenbai capsule as described in claim 1, it is characterised in that：The drug detects in this way：
(1) character：Hard capsule content is the particle and powder of sepia；Bitter；
(2) differentiate：Include thin layer discriminating, the thin layer discriminating of Radix Angelicae Sinensis, the thin layer discriminating of kuh-seng, the thin layer discriminating of Radix Astragali to jujube
And/or the thin layer of Herba Epimedii differentiates；
The thin layer of the jujube differentiates：Drug granule content 3g is taken, ethyl acetate 30ml is added, is ultrasonically treated 30 minutes, mistake
Filter, is evaporated filtrate, residue adds methanol 0.5ml to make dissolving, as test solution；Jujube control medicinal material 2g separately is taken, adds water 50ml,
It is heated to reflux 2 hours, filters while hot, filtrate volatilizes moisture and jujube thick paste is made, and cream adds ethyl acetate 30ml, is ultrasonically treated 30 points
Clock, filtering, filtrate are evaporated, and add methanol 1ml to make dissolving, as a contrast medicinal material solution, draw 5 μ l of test solution and control medicinal material
10 μ l of solution are put respectively on same silica gel H lamellae, with toluene-ethyl acetate-glacial acetic acid=60:6:0.05 is solvent,
Expansion is taken out, is dried, and spray is heated to that spot development is clear, and it is 365nm purple to set wavelength at 105 DEG C with 10% ethanol solution of sulfuric acid
It is inspected under outer lamp, in test sample chromatography, on position corresponding with reference substance chromatography, shows the fluorescence spot of same color；
The thin-layer identification method of the Radix Angelicae Sinensis is：Drug granule content 5g is taken, n-hexane 20ml is added, is ultrasonically treated 20 minutes,
Filtration, filtrate is waved to 1ml, as test solution；Radix Angelicae Sinensis control medicinal material 0.5g separately is taken, is made in the same way of control medicinal material solution；According to
Thin-layered chromatography (one VI B of annex of Chinese Pharmacopoeia version in 2010) is tested, and is drawn each 5 μ l of above two solution, is put respectively in same
On one silica gel g thin-layer plate, with n-hexane-ethyl acetate=9:1 is solvent, is unfolded, and takes out, dries, and it is 365nm purple to set wavelength
It inspects under outer lamp, in test sample chromatography, is on the corresponding position of control medicinal material chromatography, the fluorescence spot of first same color；
The thin-layer identification method of the kuh-seng is：Drug granule content 1g is taken, ethyl alcohol 30ml is added, is heated to reflux 30 minutes, puts
Cold, filtration, filtrate is evaporated residue and water 20ml is added to make dissolving, filters, and filtrate is set in separatory funnel, adds a concentration of 25%-28%'s
Strong ammonia solution 0.5ml is merged chloroform liquid, is evaporated, residue adds anhydrous second with chloroform shaking extraction 2 times, each 10ml
Alcohol 1ml makes dissolving, as test solution；Matrine and Sophoridine reference substance separately are taken, respectively plus absolute ethyl alcohol is made every 1ml and contains
The solution of 1mg, product solution, is tested according to thin-layered chromatography (one VI B of annex of Chinese Pharmacopoeia version in 2010) as a contrast, in absorption
Each 5 μ l of three kinds of solution are stated, are put respectively on same silica gel g thin-layer plate, with cyclohexane-acetone-ethyl acetate-strong ammonia solution=2:
3:4:0.5 is solvent, is set in the pre-saturated exhibition cylinder of ammonia steam, presaturation 30 minutes, is unfolded, and takes out, dries, and is sprayed with dilute iodate
Bismuth potassium test solution；In test sample chromatography, it is on the corresponding position of reference substance chromatography, shows the spot of same color；
The thin-layer identification method of the Radix Astragali is：Drug granule content 5g is taken, adds 60-90 DEG C of petroleum ether 30ml, at ultrasound
Reason 20 minutes, filtration, the dregs of a decoction wave most petroleum ether, add methanol 50ml, are heated to reflux 30 minutes, filter, and filter residue is washed with a small amount of methanol
It washs, washing lotion merges with filtrate, is evaporated, and residue adds water 10ml, heating to make dissolving, let cool, set in centrifuge tube and centrifuge 10 minutes, take
Clear liquid, with chloroform shaking extraction 2 times, each 15ml discards chloroform liquid, shakes and extracts to water saturated n-butanol
3 times, add 10ml for the first time, 10ml, third time plus 5ml, merging n-butanol liquid is added to ammoniate test solution washing 3 times, every time for the second time
15ml discards ammoniacal liquor；N-butanol liquid is washed to neutrality to what n-butanol was saturated, discards aqueous, and n-butanol liquid is evaporated, and residue adds
Methanol 1ml makes dissolving, as test solution；Astragaloside IV reference substance separately is taken, adds methanol that solution of every 1ml containing 1mg is made, is made
For reference substance solution；It is tested according to thin-layered chromatography (one VI B of annex of Chinese Pharmacopoeia version in 2010), draws 10 μ of test solution
L, 5 μ l of reference substance solution are put respectively on same silica gel g thin-layer plate, with chloroform-methanol-water=13:7:2,10 DEG C or less
Lower layer's solution of placement is solvent, is unfolded, and takes out, dries, and sprays with 10% ethanol solution of sulfuric acid, spot is heated at 105 DEG C
Colour developing is clear；In test sample chromatography, on position corresponding with reference substance chromatography, the spot of same color is shown；Setting wavelength again is
It is inspected under 365nm ultraviolet lamps, in test sample chromatography, on position corresponding with reference substance chromatography, shows the fluorescence of same color
Spot；
The thin-layer identification method of Herba Epimedii is：Drug granule content 5g is taken, 60-90 DEG C of petroleum ether 30ml is added, is ultrasonically treated
20 minutes, filtration, the dregs of a decoction waved most petroleum ether, add methanol 50ml, are heated to reflux 30 minutes, filtered, and filter residue is washed with 5ml methanol,
Washing lotion merges with filtrate, is evaporated, and residue adds water 10ml, heating to make dissolving, let cool, set in centrifuge tube and centrifuge 10 minutes, take supernatant
Liquid, with chloroform shaking extraction 2 times, each 15ml discards chloroform liquid, to water saturated n-butanol shaking extraction 3
It is secondary, add 10ml for the first time, 10ml, third time plus 5ml, merging n-butanol liquid is added to ammoniate test solution washing 3 times, every time for the second time
15ml discards ammoniacal liquor；N-butanol liquid is washed to neutrality to what n-butanol was saturated, discards aqueous, and n-butanol liquid is evaporated, and residue adds
Methanol 1ml makes dissolving, as test solution；Icariin reference substance is taken, adds methanol that solution of every 1ml containing 1mg is made, as
Reference substance solution；It is tested according to thin-layered chromatography (one VI B of annex of Chinese Pharmacopoeia version in 2010), draws 2~4 μ l of test solution
With 5 μ l of reference substance solution, put respectively on same silica gel g thin-layer plate, with chloroform-methanol-water=13:7:2,10 DEG C or less
Lower layer's solution of placement is solvent, is unfolded, and takes out, dries, and sprays with 10% ethanol solution of sulfuric acid, spot is heated at 105 DEG C
Colour developing is clear；In test sample chromatography, on position corresponding with reference substance chromatography, the spot of same color is shown；Setting wavelength again is
It is inspected under 365nm ultraviolet lamps, in test sample chromatography, on position corresponding with reference substance chromatography, shows the fluorescence of same color
Spot；
(3) assay：It include the assay to matrine and/or icariin；
The content assaying method of the matrine is：It is surveyed according to high performance liquid chromatography (four 0512 methods of Chinese Pharmacopoeia version in 2015)
It is fixed：
Chromatographic condition and system suitability：Using octadecylsilane chemically bonded silica as filler；Mobile phase：Methanol is A phases,
0.1% triethylamine is B phases, elution requirement：53%A-47%B；Detection wavelength is 220nm；Theoretical cam curve is based on matrine peak
4000 should be not less than by calculating；
The preparation of reference substance solution：Precision weighs matrine reference substance, adds methanol that solution of every 1ml containing 0.2mg is made, shakes up,
To obtain the final product；
The preparation of test solution：Drug granule content 1.0g is taken, it is accurately weighed, it sets in conical flask with cover, is added a concentration of
The strong ammonia solution 2ml wettings of 25%-28%, are added chloroform 30ml, are heated to reflux 1.5 hours, filter, filtrate is evaporated, residual
Slag add 10ml methanol dissolve, shake up to get；
Measuring method：Draw reference substance solution and each 5 μ l of test solution respectively, inject liquid chromatograph, measure to get；
The content assaying method of the icariin is：According to high performance liquid chromatography (Chinese Pharmacopoeia one annex VI of version in 2010
D it) measures：
Chromatographic condition and system suitability：Using octadecylsilane chemically bonded silica as filler；Mobile phase：Acetonitrile is A phases,
Water is B phases, elution requirement：25.5%A-74.5%B；Detection wavelength is 270nm；Theoretical cam curve is answered by the calculating of icariin peak
Not less than 3000；
The preparation of reference substance solution：Icariin reference substance is taken, it is accurately weighed, add methanol that the solution that every 1ml contains 25 μ g is made, i.e.,
?；
The preparation of test solution：The drug substance contents under content uniformity item are taken, it is finely ground, 1.5g is taken, it is accurately weighed, set tool plug cone
In shape bottle, 70% ethyl alcohol 25ml, close plug is added in precision, and weighed weight is ultrasonically treated, 1 hour, ultrasonic power 250W, supersonic frequency
Rate 35kHz takes out, lets cool, then weighed weight, the weight of less loss is supplied with 70% ethyl alcohol, is shaken up, and filters, takes subsequent filtrate, i.e.,
?；
Measuring method：It is accurate respectively to draw reference substance solution and each 10 μ l of test solution, inject liquid chromatograph, measure to get.
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