CN109030694A - The method of the finger-print identification subprostrate sophora true and false - Google Patents
The method of the finger-print identification subprostrate sophora true and false Download PDFInfo
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- CN109030694A CN109030694A CN201811102471.8A CN201811102471A CN109030694A CN 109030694 A CN109030694 A CN 109030694A CN 201811102471 A CN201811102471 A CN 201811102471A CN 109030694 A CN109030694 A CN 109030694A
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- subprostrate sophora
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- 241000219784 Sophora Species 0.000 title claims abstract description 81
- 238000000034 method Methods 0.000 title claims abstract description 23
- 239000000463 material Substances 0.000 claims abstract description 44
- 238000001514 detection method Methods 0.000 claims abstract description 11
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 32
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 30
- 239000000243 solution Substances 0.000 claims description 29
- HKQYGTCOTHHOMP-UHFFFAOYSA-N formononetin Chemical compound C1=CC(OC)=CC=C1C1=COC2=CC(O)=CC=C2C1=O HKQYGTCOTHHOMP-UHFFFAOYSA-N 0.000 claims description 24
- 230000014759 maintenance of location Effects 0.000 claims description 17
- 239000007788 liquid Substances 0.000 claims description 14
- RIKPNWPEMPODJD-UHFFFAOYSA-N formononetin Natural products C1=CC(OC)=CC=C1C1=COC2=CC=CC=C2C1=O RIKPNWPEMPODJD-UHFFFAOYSA-N 0.000 claims description 12
- 239000012071 phase Substances 0.000 claims description 11
- 239000000047 product Substances 0.000 claims description 11
- 239000012085 test solution Substances 0.000 claims description 11
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 9
- 229930013930 alkaloid Natural products 0.000 claims description 9
- 150000003797 alkaloid derivatives Chemical class 0.000 claims description 9
- 238000010828 elution Methods 0.000 claims description 9
- 238000005259 measurement Methods 0.000 claims description 9
- 150000001875 compounds Chemical class 0.000 claims description 7
- QZVQYTIOGPCCRU-JOCHJYFZSA-N sophoranone Natural products COc1c(O)c(OC)c(cc1CC=C(C)C)[C@]2(O)COc3cc(O)cc(O)c3C2=O QZVQYTIOGPCCRU-JOCHJYFZSA-N 0.000 claims description 7
- HUKSJTUUSUGIDC-ZBEGNZNMSA-N (-)-maackiain Chemical compound O1C2=CC=3OCOC=3C=C2[C@H]2[C@@H]1C1=CC=C(O)C=C1OC2 HUKSJTUUSUGIDC-ZBEGNZNMSA-N 0.000 claims description 6
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 6
- 239000000287 crude extract Substances 0.000 claims description 6
- 239000000284 extract Substances 0.000 claims description 6
- 239000000706 filtrate Substances 0.000 claims description 6
- 239000000843 powder Substances 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- VGSYCWGXBYZLLE-QEEQPWONSA-N Trifolirhizin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC=C([C@H]2[C@H](C3=CC=4OCOC=4C=C3O2)CO2)C2=C1 VGSYCWGXBYZLLE-QEEQPWONSA-N 0.000 claims description 5
- FCRUGSNPZLERNO-UHFFFAOYSA-N Trifolirhizin Natural products OCC1OC(Oc2ccc3C4Oc5c6OCOc6ccc5C4COc3c2)C(O)C(O)C1O FCRUGSNPZLERNO-UHFFFAOYSA-N 0.000 claims description 5
- 238000001914 filtration Methods 0.000 claims description 5
- VGSYCWGXBYZLLE-UHFFFAOYSA-N maackiain 3-O-beta-D-galactopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1=CC=C(C2C(C3=CC=4OCOC=4C=C3O2)CO2)C2=C1 VGSYCWGXBYZLLE-UHFFFAOYSA-N 0.000 claims description 5
- 239000003643 water by type Substances 0.000 claims description 5
- 239000007791 liquid phase Substances 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims description 4
- -1 2- (2', 4'- dihydroxy Phenyl) -5,6- dioxymethylene benzofuran Chemical class 0.000 claims description 3
- XCDMHEXDCIXKLK-UHFFFAOYSA-N Anhydrosophorol Natural products O1C2=CC=3OCOC=3C=C2C2=C1C1=CC=C(O)C=C1OC2 XCDMHEXDCIXKLK-UHFFFAOYSA-N 0.000 claims description 3
- YLZYAUCOYZKLMA-UHFFFAOYSA-N O-Methyl-maackiain Natural products O1C2=CC=3OCOC=3C=C2C2C1C1=CC=C(OC)C=C1OC2 YLZYAUCOYZKLMA-UHFFFAOYSA-N 0.000 claims description 3
- VQYBAEAOOJBSTR-QHSBEEBCSA-N Sophoranol Chemical compound C([C@@H]12)CCC(=O)N1C[C@@]1(O)[C@H]3[C@@H]2CCCN3CCC1 VQYBAEAOOJBSTR-QHSBEEBCSA-N 0.000 claims description 3
- VQYBAEAOOJBSTR-AYRXBEOTSA-N Sophoranol Natural products O=C1N2[C@H]([C@@H]3[C@@H]4[C@](O)(C2)CCCN4CCC3)CCC1 VQYBAEAOOJBSTR-AYRXBEOTSA-N 0.000 claims description 3
- HUKSJTUUSUGIDC-UHFFFAOYSA-N Trifolirhizin-aglykon Natural products O1C2=CC=3OCOC=3C=C2C2C1C1=CC=C(O)C=C1OC2 HUKSJTUUSUGIDC-UHFFFAOYSA-N 0.000 claims description 3
- 239000012982 microporous membrane Substances 0.000 claims description 3
- 238000004064 recycling Methods 0.000 claims description 3
- 238000010992 reflux Methods 0.000 claims description 3
- 239000011347 resin Substances 0.000 claims description 3
- 229920005989 resin Polymers 0.000 claims description 3
- 239000000945 filler Substances 0.000 claims description 2
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims description 2
- 239000000741 silica gel Substances 0.000 claims description 2
- 229910002027 silica gel Inorganic materials 0.000 claims description 2
- 239000000377 silicon dioxide Substances 0.000 claims description 2
- 239000000126 substance Substances 0.000 abstract description 7
- 238000012850 discrimination method Methods 0.000 abstract description 2
- 238000012544 monitoring process Methods 0.000 abstract description 2
- 238000012360 testing method Methods 0.000 description 10
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 6
- 244000046052 Phaseolus vulgaris Species 0.000 description 6
- 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 4
- ZSBXGIUJOOQZMP-UHFFFAOYSA-N Isomatrine Natural products C1CCC2CN3C(=O)CCCC3C3C2N1CCC3 ZSBXGIUJOOQZMP-UHFFFAOYSA-N 0.000 description 4
- 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 description 4
- 229930014456 matrine Natural products 0.000 description 4
- 229930015582 oxymatrine Natural products 0.000 description 4
- 238000011156 evaluation Methods 0.000 description 3
- 239000003814 drug Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 238000013112 stability test Methods 0.000 description 2
- 239000012086 standard solution Substances 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- AQASRZOCERRGBL-UHFFFAOYSA-N Dauricine Natural products CN1CCC2=CC(OC)=C(OC)C=C2C1CC1=CC=C(O)C(OC2=CC=C(C=C2)CC2N(C)CCC=3C=C(C(=CC=32)OC)OC)=C1 AQASRZOCERRGBL-UHFFFAOYSA-N 0.000 description 1
- 208000002454 Nasopharyngeal Carcinoma Diseases 0.000 description 1
- 206010061306 Nasopharyngeal cancer Diseases 0.000 description 1
- 206010068319 Oropharyngeal pain Diseases 0.000 description 1
- 201000007100 Pharyngitis Diseases 0.000 description 1
- AAGFPTSOPGCENQ-UHFFFAOYSA-N Sophocarpin I Natural products C1CCC2CN3C(=O)C=CCC3C3C2N1CCC3 AAGFPTSOPGCENQ-UHFFFAOYSA-N 0.000 description 1
- IGXQFUGORDJEST-UHFFFAOYSA-N Sophocarpine Natural products O=C1C=CCC2C3CCCC4CCCC(CN12)C34 IGXQFUGORDJEST-UHFFFAOYSA-N 0.000 description 1
- 241000123725 Sophora tonkinensis Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229940076810 beta sitosterol Drugs 0.000 description 1
- LGJMUZUPVCAVPU-UHFFFAOYSA-N beta-Sitostanol Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(C)CCC(CC)C(C)C)C1(C)CC2 LGJMUZUPVCAVPU-UHFFFAOYSA-N 0.000 description 1
- NJKOMDUNNDKEAI-UHFFFAOYSA-N beta-sitosterol Natural products CCC(CCC(C)C1CCC2(C)C3CC=C4CC(O)CCC4C3CCC12C)C(C)C NJKOMDUNNDKEAI-UHFFFAOYSA-N 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- AQASRZOCERRGBL-ROJLCIKYSA-N dauricine Chemical compound CN1CCC2=CC(OC)=C(OC)C=C2[C@H]1CC1=CC=C(O)C(OC2=CC=C(C=C2)C[C@H]2N(C)CCC=3C=C(C(=CC=32)OC)OC)=C1 AQASRZOCERRGBL-ROJLCIKYSA-N 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 229930003944 flavone Natural products 0.000 description 1
- 150000002213 flavones Chemical class 0.000 description 1
- 235000011949 flavones Nutrition 0.000 description 1
- 229940045109 genistein Drugs 0.000 description 1
- TZBJGXHYKVUXJN-UHFFFAOYSA-N genistein Natural products C1=CC(O)=CC=C1C1=COC2=CC(O)=CC(O)=C2C1=O TZBJGXHYKVUXJN-UHFFFAOYSA-N 0.000 description 1
- 235000006539 genistein Nutrition 0.000 description 1
- ZCOLJUOHXJRHDI-CMWLGVBASA-N genistein 7-O-beta-D-glucoside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC(O)=C2C(=O)C(C=3C=CC(O)=CC=3)=COC2=C1 ZCOLJUOHXJRHDI-CMWLGVBASA-N 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- 150000002338 glycosides Chemical class 0.000 description 1
- 239000002398 materia medica Substances 0.000 description 1
- 239000012567 medical material Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 201000011216 nasopharynx carcinoma Diseases 0.000 description 1
- 150000002825 nitriles Chemical class 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
- 230000035945 sensitivity Effects 0.000 description 1
- KZJWDPNRJALLNS-VJSFXXLFSA-N sitosterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CC[C@@H](CC)C(C)C)[C@@]1(C)CC2 KZJWDPNRJALLNS-VJSFXXLFSA-N 0.000 description 1
- 229950005143 sitosterol Drugs 0.000 description 1
- AAGFPTSOPGCENQ-JLNYLFASSA-N sophocarpine Chemical compound C1CC[C@H]2CN3C(=O)C=CC[C@@H]3[C@@H]3[C@H]2N1CCC3 AAGFPTSOPGCENQ-JLNYLFASSA-N 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 238000010025 steaming Methods 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 229940126680 traditional chinese medicines Drugs 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
- G01N30/86—Signal analysis
- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
- G01N30/8686—Fingerprinting, e.g. without prior knowledge of the sample components
Landscapes
- Engineering & Computer Science (AREA)
- Library & Information Science (AREA)
- 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)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a kind of methods that finger-print identifies the subprostrate sophora true and false, judge the true and false of the subprostrate sophora medicinal material sample by measuring the chromatograms of subprostrate sophora medicinal material sample.Discrimination method provided by the invention can reflect the active chemical of subprostrate sophora comprehensively, reflect the quality of subprostrate sophora medicinal material sample on the whole, accurately and reliably, and it is easy to operation, at low cost, practical, be conducive to promote on a large scale, be highly suitable for the detection and monitoring of subprostrate sophora medicinal material and products thereof quality.
Description
Technical field
The present invention relates to a kind of methods for identifying the subprostrate sophora true and false.It is more particularly related to a kind of finger-print
The method for identifying the subprostrate sophora true and false, belongs to Materia Medica Identification method and technology field.
Background technique
Subprostrate sophora is the drying root and rhizome of leguminous plant sophora tonkinensis Gapnep, has the effect of clearing heat and detoxicating, relieving sore-throat of reducing swelling, contains
The ingredients such as matrine, oxymatrine, sophocarpine, Dauricine, genistein, β-sitosterol, flavones, existing " middle traditional Chinese medicines
Allusion quotation " it is fairly simple to subprostrate sophora method of quality control, only using matrine and oxymatrine concentration as evaluation of medical materials' quality
Index, and matrine and oxymatrine are not subprostrate sophora endemic elements, and to the index determining of matrine and oxymatrine
It is difficult to react quality of medicinal material on the whole.
A kind of effective mass control method of the fingerprint pattern technology as Multi-chemical ingredients sample, can be on the whole
The Integral Characteristic for reflecting sample to be tested is widely used in the control of Chinese medicine quality at present.So for more acurrate evaluation mountain
Beans root herb quality needs to establish a kind of finger-print that can reflect subprostrate sophora active chemical comprehensively.
Summary of the invention
It is an object of the present invention to provide a kind of methods that finger-print identifies the subprostrate sophora true and false, can reflect mountain comprehensively
Active chemical in beans root reflects the quality of subprostrate sophora medicinal material sample on the whole.
In order to realize these purposes and other advantages according to the present invention, it is true to provide a kind of finger-print identification subprostrate sophora
Pseudo- method judges the true and false of the subprostrate sophora medicinal material sample by measuring the chromatograms of subprostrate sophora medicinal material sample.
Preferably, using 10 chromatographic peaks in standard subprostrate sophora non-alkaloid finger-print as characteristic peak, when mountain beans
When there are not this 10 characteristic peaks simultaneously in the chromatograms of root herb sample, subprostrate sophora medicinal material sample is fake and poor products；When mountain beans
The relative retention time of 10 characteristic peaks and standard subprostrate sophora non-alkaloid finger-print in the chromatograms of root herb sample
When similarity is higher than 85%, subprostrate sophora medicinal material sample is high-quality product, wherein by peak sequence with the third in 10 characteristic peaks
Characteristic peak is referring to peak, the relative retention time of remaining 9 characteristic peak is respectively as follows: 0.403,0.462,1.077,1.600,
1.883、1.899、1.948、1.975、2.081。
It preferably, is respectively trifolirhizin, different screw oil expeller axis by the corresponding compound of peak sequence first four characteristic peak
Careless glycosides, formononetin, 2- (2', 4'- dihydroxy phenyl) -5,6- dioxymethylene benzofuran, the 6th characteristic peak are corresponding
Compound be 3- methoxyl group maackiain, the 9th corresponding compound of characteristic peak is Vietnam's sophoranol.
Preferably, test solution first is made in subprostrate sophora medicinal material sample, then measures the liquid phase figure of test solution
Spectrum, wherein the preparation process of test solution specifically includes: weighing subprostrate sophora medicinal material sample powder, and subprostrate sophora medicinal material sample is added
The extracting solution of 5~20 times of weight of product powder, 1~2h of refluxing extraction, filtering take filtrate, and filtrate is carried out vacuum rotary steam recycling and is mentioned
It takes liquid to being evaporated, obtains crude extract, be dissolved in water into crude extract, cross macroreticular resin, be 10~39% with volume fraction
After ethanol solution elution, then the ethanol solution for being 40~95% with volume fraction elutes, and the ethanol solution for collecting 40~95% is washed
De- part, vacuum rotary steam recycle ethanol solution to being evaporated, obtain essence extract, and into essence extract plus methanol dissolves, with 0.45 μm
After filtering with microporous membrane, constant volume.
Preferably, extracting solution is the ethanol solution that volume fraction is 65~95%.
Preferably, the actual conditions for measuring the chromatograms of subprostrate sophora medicinal material sample include: high performance liquid chromatograph institute
For chromatographic column using octadecylsilane chemically bonded silica as filler, column temperature is 25~35 DEG C；Wavelength is when liquid chromatographic detection
200~320nm, sample volume are 5~10 μ l, and flow velocity is 0.8~1.2ml/min；Liquid chromatographic detection takes gradient elution,
In, acetonitrile is mobile phase A, and water is Mobile phase B, the variation of volume ratio shared by acetonitrile specifically: in 0~5min, shared by acetonitrile
Volume ratio is 25%, and in 5~55min, volume ratio shared by acetonitrile is gradually increased from 25% to 50%, in 55~80min, second
Volume ratio shared by nitrile is gradually increased from 50% to 95%.
Preferably, the actual conditions for measuring the chromatograms of subprostrate sophora medicinal material sample include: high performance liquid chromatograph institute
Chromatographic column is Waters Sunfire-C18 column, and column internal diameter 4.6mm, pillar height 250mm, silica gel partial size is 5 μm, column temperature
It is 30 DEG C；Wavelength is 205nm when liquid chromatographic detection, and sample volume is 10 μ l, flow velocity 1.0ml/min.
The present invention is include at least the following beneficial effects: the present invention passes through the chromatograms of measurement subprostrate sophora medicinal material sample, so
It is compared afterwards with standard subprostrate sophora non-alkaloid finger-print, using 10 characteristic peaks as the evaluation index of subprostrate sophora quality, energy
The active chemical of reflection subprostrate sophora comprehensively, reflects the quality of subprostrate sophora medicinal material on the whole, can accurately and reliably identify mountain
The true and false of beans root herb has the advantages that medicinal material sample treatment is simple and convenient, testing result is reproducible, detection sensitivity is high,
It is able to achieve the stability contorting to subprostrate sophora medicinal material and products thereof quality, guarantees the security provisions of subprostrate sophora medicinal material and products thereof, has
Conducive to wideling popularize and apply subprostrate sophora.The present invention is tried by similarity test, precision test, stability test, repeatability
It tests to point out to test with characteristic peak and this method is verified, the results showed that detection method provided by the invention is accurate and reliable, can
Reflect multiple active chemicals in subprostrate sophora, reflect the quality of subprostrate sophora medicinal material on the whole, so that acquisition is more accurate
Reliable testing result.In addition, discrimination method provided by the invention is easy to operation, at low cost, practical, it is suitble to big model
Popularization is enclosed, the detection and monitoring of subprostrate sophora medicinal material and products thereof quality are highly suitable for.
Further advantage, target and feature of the invention will be partially reflected by the following instructions, and part will also be by this
The research and practice of invention and be understood by the person skilled in the art.
Detailed description of the invention
Fig. 1 is the chromatograms of 10 batches of subprostrate sophora genuine pieces described in a wherein embodiment of the invention；
Fig. 2 is standard subprostrate sophora non-alkaloid finger-print described in a wherein embodiment of the invention.
Specific embodiment
The present invention is described in further detail with reference to the accompanying drawings and examples, to enable those skilled in the art's reference
Specification word can be implemented accordingly.
It should be noted that experimental method described in following embodiments is unless otherwise specified conventional method, institute
Reagent and material are stated, unless otherwise specified, is commercially obtained.
<embodiment>
Finger-print identify the subprostrate sophora true and false method the following steps are included:
Step 1: weighing subprostrate sophora medicinal material sample powder 2g, and 20 times of weight of subprostrate sophora medicinal material sample powder, volume point is added
Number is 85% ethanol solution, and refluxing extraction 2h, filtering takes filtrate, by filtrate progress vacuum rotary steam recycling ethanol solution to steaming
It is dry, crude extract is obtained, into crude extract plus 20ml water dissolves, and crosses macroreticular resin, the ethanol solution for being 39% with volume fraction
After elution, then the ethanol solution for being 95% with volume fraction elutes, and collects 95% ethanol solution elution fraction, and vacuum rotary steam returns
Ethanol solution is received to being evaporated, obtains essence extract, into essence extract plus methanol dissolution is determined after 0.45 μm of filtering with microporous membrane
Hold to 10ml volumetric flask, obtains test solution；
Step 2: the chromatograms of test solution are measured, wherein the actual conditions for measuring chromatograms include:
Chromatograph: 2695 high performance liquid chromatograph of Waters (Waters, US)；
Chromatographic column: Waters Sunfire-C18 (4.6mm × 250mm, 5 μm) chromatographic column；
Column temperature: 30 DEG C；
Detection wavelength: 205nm；
Sample volume: 10 μ l；
Flow velocity: 1.0ml/min；
Mobile phase: acetonitrile is mobile phase A, and water is Mobile phase B, carries out gradient elution by table 1:
[table 1]
Step 3: by 10 in the chromatograms of subprostrate sophora medicinal material sample and standard subprostrate sophora non-alkaloid finger-print
Characteristic peak is compared, when there are not this 10 characteristic peaks simultaneously in the chromatograms of subprostrate sophora medicinal material sample, subprostrate sophora medicine
Material sample is fake and poor products；When the relative retention time of 10 characteristic peaks in the chromatograms of subprostrate sophora medicinal material sample and standard mountain beans
When the similarity of root non-alkaloid finger-print is higher than 85%, subprostrate sophora medicinal material sample is high-quality product.
Wherein, 10 characteristic peaks are respectively labeled as 1~No. 10 peak by peak sequence, are referring to peak with No. 3 peaks, remaining 9
The relative retention time of a characteristic peak is respectively as follows: 0.403,0.462,1.077,1.600,1.883,1.899,1.948,1.975,
2.081。
<similarity test>
10 batches of subprostrate sophora genuine pieces (source is shown in Table 2) are taken, according to the preparation method of test solution in embodiment by subprostrate sophora
Genuine piece solution is made in genuine piece, and measures 10 batches of subprostrate sophora genuine piece solution using chromatograms determination condition identical with embodiment
Chromatograms, and record the chromatograms of this 10 batches of subprostrate sophora genuine pieces, the result is shown in Figure 1.
Wherein, the chromatograms separating degree and peak shape of No. 3 subprostrate sophora genuine pieces are best, as standard subprostrate sophora non-alkaloid
Finger-print is shown in Fig. 2, and 1~10 marked in Fig. 2 is 10 selected characteristic peaks, for the liquid with subprostrate sophora medicinal material sample
Phase map is compared, to identify the true and false of subprostrate sophora medicinal material sample.The retention time of 10 characteristic peaks is respectively as follows: 15.291,
17.562,37.976,40.894,60.753,71.513,72.119,73.993,75.003,79.046 (unit: min).Due to
Formononetin occurs in each batch of subprostrate sophora genuine piece, and separating degree is good, therefore formononetin (i.e. No. 3 peaks) is selected to make
For referring to peak (be labeled as S), the relative retention time of remaining 9 characteristic peak is respectively as follows: 0.403,0.462,1.077,1.600,
1.883、1.899、1.948、1.975、2.081。
Formononetin standard specimen is taken, standard solution identical with genuine piece solution concentration, and use and embodiment are configured to
The chromatograms of identical chromatograms determination condition measurement formononetin standard solution, obtain the liquid of formononetin standard specimen
Phase map calculates 10 batches of subprostrate sophoras on the basis of the retention time of formononetin in the chromatograms of formononetin standard specimen
Referring to the relative retention time (being denoted as S1~S10) of peak (S) in the chromatograms of genuine piece, it the results are shown in Table 3.From table 3, it can be seen that
S1~S10 illustrates the reservation at formononetin peak in the chromatograms of 10 batches of subprostrate sophora genuine pieces between 0.893~0.981
The similarity of the retention time of time and formononetin standard specimen is between 0.893~0.981, and similarity is very high, and explanation is adopted
The true and false for identifying subprostrate sophora medicinal material sample with liquid-phase fingerprint is to compare accurately and reliably.
[table 2]
[table 3]
<precision test>
It takes genuine piece solution continuous sample introduction 6 times with a subprostrate sophora genuine piece, is surveyed using chromatograms identical with embodiment
Fixed condition is measured, the chromatograms after recording measurement every time, is to calculate the phase of remaining 9 characteristic peak referring to peak with No. 3 peaks
To retention time and relative peak area, as the result is shown: RSD (the i.e. relative standard of each characteristic peak relative retention time in 6 measurements
Deviation) < 0.12%, RSD < 2.93% of relative peak area shows that method precision provided by the invention is good.
<stability test>
Take the genuine piece solution with a subprostrate sophora genuine piece, respectively 0,2,4,8,12, measure its chromatograms, liquid phase for 24 hours
The determination condition of map is identical as embodiment, the chromatograms after recording measurement every time, with No. 3 peaks be referring to peak, calculate remaining 9
The relative retention time and relative peak area of a characteristic peak, as the result is shown: for 24 hours in each characteristic peak relative retention time RSD <
0.31%, RSD < 2.04% of relative peak area shows that test solution stablizes interior for 24 hours.
<repetitive test>
It takes with a collection of 6 parts of genuine piece of subprostrate sophora, according to the preparation method of test solution in embodiment by subprostrate sophora genuine piece system
At genuine piece solution, and using the chromatograms of chromatograms determination condition identical with embodiment 6 parts of genuine piece solution of measurement, record
Chromatograms after measurement every time are the relative retention time that remaining 9 characteristic peak is calculated referring to peak and opposite peak with No. 3 peaks
Area, as the result is shown: RSD < 0.15% of each characteristic peak relative retention time in 6 parts of genuine piece solution, the RSD of relative peak area <
2.94%, show method repeatability provided by the invention preferably.
<characteristic peak points out test>
By the single comparison of a variety of standard specimens, identifying has the corresponding compound of 6 characteristic peaks in 10 characteristic peaks are as follows: 1
Number peak is trifolirhizin, and No. 2 peaks are different trifolirhizin, and No. 3 peaks are formononetin, and No. 4 peaks are 2- (2', 4'- dihydroxy
Phenyl) -5,6- dioxymethylene benzofuran, No. 6 peaks are 3- methoxyl group maackiain, and No. 9 peaks are Vietnam's sophoranol.This changes in 6
The active constituent that object is the anti-nasopharyngeal carcinoma of subprostrate sophora is closed, illustrates that method provided by the invention can preferably reflect the effective of subprostrate sophora
Chemical component.
Although the embodiments of the present invention have been disclosed as above, but its is not only in the description and the implementation listed
With it can be fully applied to various fields suitable for the present invention, for those skilled in the art, can be easily
Realize other modification, therefore without departing from the general concept defined in the claims and the equivalent scope, the present invention is simultaneously unlimited
In specific details and example shown and described herein and legend.
Claims (7)
1. the method for the finger-print identification subprostrate sophora true and false, which is characterized in that by the liquid phase figure for measuring subprostrate sophora medicinal material sample
Compose the true and false to judge the subprostrate sophora medicinal material sample.
2. the method that finger-print as described in claim 1 identifies the subprostrate sophora true and false, which is characterized in that non-with standard subprostrate sophora
10 chromatographic peaks in fingerprint of alkaloid are as characteristic peak, when not occurring simultaneously in the chromatograms of subprostrate sophora medicinal material sample
When this 10 characteristic peaks, subprostrate sophora medicinal material sample is fake and poor products；When 10 characteristic peaks in the chromatograms of subprostrate sophora medicinal material sample
Relative retention time and standard subprostrate sophora non-alkaloid finger-print similarity be higher than 85% when, subprostrate sophora medicinal material sample is
High-quality product, wherein by peak sequence with the third characteristic peak in 10 characteristic peaks for referring to peak, the phase of remaining 9 characteristic peak
0.403,0.462,1.077,1.600,1.883,1.899,1.948,1.975,2.081 are respectively as follows: to retention time.
3. the method that finger-print as claimed in claim 2 identifies the subprostrate sophora true and false, which is characterized in that by before peak sequence four
The corresponding compound of a characteristic peak is respectively trifolirhizin, different trifolirhizin, formononetin, 2- (2', 4'- dihydroxy
Phenyl) -5,6- dioxymethylene benzofuran, the 6th corresponding compound of characteristic peak is 3- methoxyl group maackiain, the 9th
The corresponding compound of a characteristic peak is Vietnam's sophoranol.
4. the method that finger-print as described in claim 1 identifies the subprostrate sophora true and false, which is characterized in that first by subprostrate sophora medicinal material
Test solution is made in sample, then measures the chromatograms of test solution, wherein the preparation process of test solution is specific
Include: to weigh subprostrate sophora medicinal material sample powder, the extracting solution of 5~20 times of weight of subprostrate sophora medicinal material sample powder is added, reflux mentions
1~2h is taken, filters, takes filtrate, filtrate is subjected to vacuum rotary steam recycling extracting solution to being evaporated, crude extract is obtained, to crude extract
In be dissolved in water, cross macroreticular resin, with volume fraction be 10~39% ethanol solution elution after, then with volume fraction for 40~
95% ethanol solution elution, collects 40~95% ethanol solution elution fraction, and vacuum rotary steam recycles ethanol solution to being evaporated,
Essence extract is obtained, into essence extract plus methanol dissolves, after 0.45 μm of filtering with microporous membrane, constant volume.
5. the method that finger-print as claimed in claim 4 identifies the subprostrate sophora true and false, which is characterized in that extracting solution is volume point
The ethanol solution that number is 65~95%.
6. the method that finger-print as described in claim 1 identifies the subprostrate sophora true and false, which is characterized in that measurement subprostrate sophora medicinal material
The actual conditions of the chromatograms of sample include: chromatographic column used in high performance liquid chromatograph with octadecylsilane chemically bonded silica
For filler, column temperature is 25~35 DEG C；Wavelength is 200~320nm when liquid chromatographic detection, and sample volume is 5~10 μ l, and flow velocity is
0.8~1.2ml/min；Liquid chromatographic detection takes gradient elution, wherein acetonitrile is mobile phase A, and water is Mobile phase B, acetonitrile institute
Account for the variation of volume ratio specifically: in 0~5min, volume ratio shared by acetonitrile is 25%, in 5~55min, body shared by acetonitrile
Product ratio is gradually increased from 25% to 50%, and in 55~80min, volume ratio shared by acetonitrile is gradually increased from 50% to 95%.
7. the method that finger-print as claimed in claim 6 identifies the subprostrate sophora true and false, which is characterized in that measurement subprostrate sophora medicinal material
The actual conditions of the chromatograms of sample include: that chromatographic column used in high performance liquid chromatograph is Waters Sunfire-C18
Column, column internal diameter 4.6mm, pillar height 250mm, silica gel partial size are 5 μm, and column temperature is 30 DEG C；Wavelength is when liquid chromatographic detection
205nm, sample volume are 10 μ l, flow velocity 1.0ml/min.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111317741A (en) * | 2020-04-23 | 2020-06-23 | 广西壮族自治区药用植物园 | Traditional Chinese medicine oral preparation for treating nasopharyngeal carcinoma and identification method and application thereof |
| CN112505216A (en) * | 2020-09-04 | 2021-03-16 | 广西壮族自治区药用植物园 | One-test-multiple-evaluation method for three non-alkaloid components in subprostrate sophora and application thereof |
| CN113588857A (en) * | 2021-07-28 | 2021-11-02 | 中国人民解放军海军军医大学 | Quality detection method of subprostrate sophora and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2013100236A (en) * | 2011-11-07 | 2013-05-23 | Dhc Co | Bbf2h7 expression increasing agent |
| CN103301190A (en) * | 2012-06-19 | 2013-09-18 | 中国医学科学院药用植物研究所 | Extracts of vietnamese sophora root and application thereof |
| CN103487516A (en) * | 2012-06-12 | 2014-01-01 | 河北以岭医药研究院有限公司 | Fingerprint chromatography determination method of traditional Chinese medicine composition preparation |
| CN103837619A (en) * | 2014-03-20 | 2014-06-04 | 常熟雷允上制药有限公司 | Fingerprint detection method for traditional Chinese medicinal Ganyanlin injection and raw medicinal material thereof |
-
2018
- 2018-09-20 CN CN201811102471.8A patent/CN109030694B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2013100236A (en) * | 2011-11-07 | 2013-05-23 | Dhc Co | Bbf2h7 expression increasing agent |
| CN103487516A (en) * | 2012-06-12 | 2014-01-01 | 河北以岭医药研究院有限公司 | Fingerprint chromatography determination method of traditional Chinese medicine composition preparation |
| CN103301190A (en) * | 2012-06-19 | 2013-09-18 | 中国医学科学院药用植物研究所 | Extracts of vietnamese sophora root and application thereof |
| CN103837619A (en) * | 2014-03-20 | 2014-06-04 | 常熟雷允上制药有限公司 | Fingerprint detection method for traditional Chinese medicinal Ganyanlin injection and raw medicinal material thereof |
Non-Patent Citations (4)
| Title |
|---|
| 刘吉成 等: "广西多叶越南槐HPLC指纹图谱的建立及与山豆根HPLC指纹图谱一致性比较研究", 《中国实验方剂学杂志》 * |
| 李秋萍 等: "山豆根非生物碱化学成分研究进展", 《中国现代中药》 * |
| 陈影 等: "山豆根不同提取部位的毒性作用研究", 《中国药物警戒》 * |
| 黄亚非 等: "广西不同产地山豆根的指纹图谱特征研究", 《中药材》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111317741A (en) * | 2020-04-23 | 2020-06-23 | 广西壮族自治区药用植物园 | Traditional Chinese medicine oral preparation for treating nasopharyngeal carcinoma and identification method and application thereof |
| CN112505216A (en) * | 2020-09-04 | 2021-03-16 | 广西壮族自治区药用植物园 | One-test-multiple-evaluation method for three non-alkaloid components in subprostrate sophora and application thereof |
| CN113588857A (en) * | 2021-07-28 | 2021-11-02 | 中国人民解放军海军军医大学 | Quality detection method of subprostrate sophora and application thereof |
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