CN116609448A - Construction method and identification method of characteristic patterns of lily medicinal materials with different origins and preparations thereof - Google Patents
Construction method and identification method of characteristic patterns of lily medicinal materials with different origins and preparations thereof Download PDFInfo
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- CN116609448A CN116609448A CN202310350947.4A CN202310350947A CN116609448A CN 116609448 A CN116609448 A CN 116609448A CN 202310350947 A CN202310350947 A CN 202310350947A CN 116609448 A CN116609448 A CN 116609448A
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- 241000234435 Lilium Species 0.000 title claims abstract description 328
- 239000000463 material Substances 0.000 title claims abstract description 122
- 238000000034 method Methods 0.000 title claims abstract description 80
- 238000002360 preparation method Methods 0.000 title claims abstract description 63
- 238000010276 construction Methods 0.000 title claims abstract description 20
- 239000012488 sample solution Substances 0.000 claims abstract description 52
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims abstract description 51
- 238000010828 elution Methods 0.000 claims abstract description 29
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims abstract description 26
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 26
- 238000001514 detection method Methods 0.000 claims abstract description 17
- 238000004128 high performance liquid chromatography Methods 0.000 claims abstract description 17
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 15
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims abstract description 13
- 238000005096 rolling process Methods 0.000 claims abstract description 8
- 125000001295 dansyl group Chemical group [H]C1=C([H])C(N(C([H])([H])[H])C([H])([H])[H])=C2C([H])=C([H])C([H])=C(C2=C1[H])S(*)(=O)=O 0.000 claims abstract description 7
- 239000000945 filler Substances 0.000 claims abstract description 7
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims abstract description 7
- 239000000377 silicon dioxide Substances 0.000 claims abstract description 7
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 66
- 238000001228 spectrum Methods 0.000 claims description 65
- YSCJAYPKBYRXEZ-HZPINHDXSA-N (2s,3s,4s,5r,6r)-6-[[(3s,4ar,6ar,6bs,8as,12as,14ar,14br)-4,4,6a,6b,11,11,14b-heptamethyl-8a-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxycarbonyl-1,2,3,4a,5,6,7,8,9,10,12,12a,14,14a-tetradecahydropicen-3-yl]oxy]-3-hydroxy-4-[(2s,3r,4s, Chemical compound O([C@H]1[C@H](O)[C@H](O[C@H]([C@@H]1O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)O[C@H]1CC[C@]2(C)[C@H]3CC=C4[C@@]([C@@]3(CC[C@H]2C1(C)C)C)(C)CC[C@]1(CCC(C[C@H]14)(C)C)C(=O)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)C(O)=O)[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O YSCJAYPKBYRXEZ-HZPINHDXSA-N 0.000 claims description 60
- 230000014759 maintenance of location Effects 0.000 claims description 51
- 238000012360 testing method Methods 0.000 claims description 45
- 239000013558 reference substance Substances 0.000 claims description 42
- 239000000243 solution Substances 0.000 claims description 41
- 239000000523 sample Substances 0.000 claims description 32
- MOMFXATYAINJML-UHFFFAOYSA-N 2-Acetylthiazole Chemical group CC(=O)C1=NC=CS1 MOMFXATYAINJML-UHFFFAOYSA-N 0.000 claims description 31
- 229930182470 glycoside Natural products 0.000 claims description 27
- 150000002338 glycosides Chemical class 0.000 claims description 27
- 239000007788 liquid Substances 0.000 claims description 25
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 14
- 239000007864 aqueous solution Substances 0.000 claims description 12
- 239000012088 reference solution Substances 0.000 claims description 11
- 238000001914 filtration Methods 0.000 claims description 10
- 238000000605 extraction Methods 0.000 claims description 9
- 238000002347 injection Methods 0.000 claims description 8
- 239000007924 injection Substances 0.000 claims description 8
- 238000000926 separation method Methods 0.000 claims description 8
- 238000010992 reflux Methods 0.000 claims description 7
- 239000002904 solvent Substances 0.000 claims description 7
- 241000847925 Paeonia ostii Species 0.000 claims description 5
- 238000010438 heat treatment Methods 0.000 claims description 5
- 238000001035 drying Methods 0.000 claims description 3
- 239000007787 solid Substances 0.000 claims description 3
- 238000005119 centrifugation Methods 0.000 claims description 2
- 238000002137 ultrasound extraction Methods 0.000 claims description 2
- 238000003809 water extraction Methods 0.000 claims description 2
- 230000003595 spectral effect Effects 0.000 claims 1
- 239000003814 drug Substances 0.000 abstract description 16
- 239000000843 powder Substances 0.000 description 38
- 241001359444 Lilium tenuifolium Species 0.000 description 14
- 239000008176 lyophilized powder Substances 0.000 description 13
- 238000002474 experimental method Methods 0.000 description 11
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- 239000000126 substance Substances 0.000 description 6
- 241001104043 Syringa Species 0.000 description 5
- 235000004338 Syringa vulgaris Nutrition 0.000 description 5
- 238000001816 cooling Methods 0.000 description 5
- 239000006187 pill Substances 0.000 description 5
- 230000001105 regulatory effect Effects 0.000 description 5
- 238000000862 absorption spectrum Methods 0.000 description 4
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- 238000011156 evaluation Methods 0.000 description 4
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- 238000011835 investigation Methods 0.000 description 4
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- 238000001704 evaporation Methods 0.000 description 3
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- 238000004587 chromatography analysis Methods 0.000 description 2
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- 238000012545 processing Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 230000001502 supplementing effect Effects 0.000 description 2
- 229910021642 ultra pure water Inorganic materials 0.000 description 2
- 239000012498 ultrapure water Substances 0.000 description 2
- 238000012795 verification Methods 0.000 description 2
- PUAQLLVFLMYYJJ-UHFFFAOYSA-N 2-aminopropiophenone Chemical compound CC(N)C(=O)C1=CC=CC=C1 PUAQLLVFLMYYJJ-UHFFFAOYSA-N 0.000 description 1
- WOXRFOGJAFXBAH-RSDOQXBQSA-N CC(=O)OCC(CO)O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O Chemical compound CC(=O)OCC(CO)O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O WOXRFOGJAFXBAH-RSDOQXBQSA-N 0.000 description 1
- WOXRFOGJAFXBAH-UHFFFAOYSA-N Lilioside A Natural products CC(=O)OCC(CO)OC1OC(CO)C(O)C(O)C1O WOXRFOGJAFXBAH-UHFFFAOYSA-N 0.000 description 1
- 240000008058 Lilium brownii Species 0.000 description 1
- 235000015982 Lilium brownii Nutrition 0.000 description 1
- 244000210789 Lilium lancifolium Species 0.000 description 1
- 235000002156 Lilium lancifolium Nutrition 0.000 description 1
- 240000005001 Paeonia suffruticosa Species 0.000 description 1
- 235000003889 Paeonia suffruticosa Nutrition 0.000 description 1
- 235000011096 Papaver Nutrition 0.000 description 1
- 240000001090 Papaver somniferum Species 0.000 description 1
- 241001474728 Satyrodes eurydice Species 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 238000001647 drug administration Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 238000005469 granulation Methods 0.000 description 1
- 230000003179 granulation Effects 0.000 description 1
- 235000008216 herbs Nutrition 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 229930189484 lilioside Natural products 0.000 description 1
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 238000013441 quality evaluation Methods 0.000 description 1
- QEVHRUUCFGRFIF-MDEJGZGSSA-N reserpine Chemical compound O([C@H]1[C@@H]([C@H]([C@H]2C[C@@H]3C4=C(C5=CC=C(OC)C=C5N4)CCN3C[C@H]2C1)C(=O)OC)OC)C(=O)C1=CC(OC)=C(OC)C(OC)=C1 QEVHRUUCFGRFIF-MDEJGZGSSA-N 0.000 description 1
- 239000004576 sand Substances 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000004506 ultrasonic cleaning Methods 0.000 description 1
Classifications
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- 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
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- 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
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/36—Control of physical parameters of the fluid carrier in high pressure liquid systems
-
- 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
- G01N30/50—Conditioning of the sorbent material or stationary liquid
- G01N30/52—Physical parameters
- G01N30/54—Temperature
-
- 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
- G01N30/62—Detectors specially adapted therefor
- G01N30/74—Optical detectors
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- 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)
- Spectroscopy & Molecular Physics (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention belongs to the technical field of traditional Chinese medicine detection, and particularly provides a construction method and an identification method of characteristic patterns of lily medicinal materials with different origins and preparations thereof, which comprises the following steps of (1) preparing lily sample solution; (2) Detecting lily sample solution by high performance liquid chromatography, taking octadecylsilane chemically bonded silica as filler, and performing gradient elution with mobile phase comprising acetonitrile and 0.05-0.2vt% phosphoric acid water solution, wherein the gradient elution procedure comprises: the volume percentage of acetonitrile in the mobile phase is 6% -12% -18% -40% -70%, the characteristic maps of lily medicinal materials with different base sources and preparations thereof are established, and the characteristic maps can be used for identifying and distinguishing lily medicinal materials with a rolling dansyl source and preparations thereof, lily medicinal materials with a lily base source and preparations thereof and lily medicinal materials with a fine leaf lily base and preparations thereof.
Description
Technical Field
The invention belongs to the technical field of traditional Chinese medicine detection, and particularly relates to a construction method and an identification method of characteristic patterns of lily medicinal materials with different origins and preparations thereof.
Background
The "chinese pharmacopoeia" of 2020 provides that the source of lily is dried fleshy scale leaf of Lilium schoendole Lilium lancifolium thunder et al, lilium brownii f.e. brown var. The medicinal lily in the market at present mainly takes the lilium schoendole as a main component, and secondly takes the lilium schoendole as a primary component, the flow rate of the fine-leaf lily is lower, the lilium schoendole, the lilium schoendole and the lilium schoendole as a primary component has larger difference in terms of the content of effective components and the efficacy, the characteristics of lily decoction pieces from different sources are similar, no experience personnel can not accurately identify the lilium schoendole, the quality evaluation basis of the lilium schoendole in Chinese pharmacopoeia is only character thin layer identification and water-soluble extract content, no specific variety identification method is adopted, and the accuracy of drug administration is difficult to ensure, and the method is also not applicable to standard decoction or lily formula granules taking the lilium schoendole, the lilium schoendole and the fine-leaf lily as a primary component.
The traditional Chinese medicine formula granule and the standard decoction are common formulations of lily, wherein the traditional Chinese medicine formula granule is prepared by extracting traditional Chinese medicine decoction pieces with water, and by the processes of extraction, drying, granulation and the like, and the clinical curative effect of the traditional Chinese medicine formula granule is consistent with that of the decoction. The standard decoction is a material standard for measuring whether the traditional Chinese medicine formula particles are basically consistent with the decoction. However, the characteristics of the original medicinal materials are lost no matter the medicinal materials are standard decoction or Chinese medicinal granules, namely, the medicinal materials cannot be checked and identified from the shapes, the sizes, the textures and the like of the medicinal materials.
Disclosure of Invention
Therefore, the invention solves the problem that the method in the prior art cannot accurately identify the lily medicinal materials and the preparations thereof taking the lilium schoendole, the lilium schoendole and the lilium tenuifolium as the base, provides the construction method and the identification method of the characteristic patterns of the lily medicinal materials and the preparations thereof with different base, establishes the characteristic patterns of the lily medicinal materials and the preparations thereof with different base, identifies and distinguishes the lilium schoendole medicinal materials and the preparations thereof with the lilium schoendole and the lilium tenuifolium as the base, and provides a rapid and reliable detection method for the interplanetary identification of the lily medicinal materials and the preparations thereof.
The invention provides a construction method of characteristic patterns of lily medicinal materials with different basic origins and preparations thereof, which comprises the following steps:
(1) Preparing a lily test sample solution;
(2) Detecting lily sample solution by high performance liquid chromatography, taking octadecylsilane chemically bonded silica as filler, and performing gradient elution with mobile phase comprising acetonitrile and 0.05-0.2vt% phosphoric acid water solution, wherein the gradient elution procedure comprises: 0- > 10 min- > 15 min- > 23 min- > 30 min- > 35 min, the volume percentage of acetonitrile in the mobile phase is 6% → 12% → 18% → 18% → 40% → 70%.
Further, step (1) includes:
1) Extracting Bulbus Lilii sample with water to obtain extractive solution;
2) Extracting the extracting solution by adding an alcohol water solution after solid-liquid separation and drying to obtain the extracting solution;
3) Separating solid from liquid, and collecting the liquid as sample solution; alternatively, step (1) includes:
1) Extracting Bulbus Lilii sample with alcohol water solution to obtain extractive solution;
2) And (3) carrying out solid-liquid separation on the extracting solution, and taking the liquid as a sample solution.
Further, step (1) also satisfies any one or more of the following A-F:
A. the water extraction is carried out by adding water, heating and refluxing for at least 20 min, preferably 20-60min;
B. the ratio of the mass of the lily test sample to the volume of water is 0.5-2:50, and the ratio relationship of the mass to the solvent is g/mL;
C. the extraction of the alcohol aqueous solution is reflux extraction or ultrasonic extraction after adding the alcohol aqueous solution, and the extraction time is 10min-5h;
D. the alcohol aqueous solution is selected from ethanol aqueous solution or methanol aqueous solution with the volume percentage of 30-70%;
E. the solid-liquid separation is centrifugation or filtration;
F. the ratio of the mass of the lily test sample to the volume of the alcohol-water solution is 0.2-2:20, and the ratio relationship of the mass to the solvent is g/mL.
Further, in the step (2), the detection wavelength is 203-208nm before 14-20min, the detection wavelength is 308-313nm after 14-20min, the flow rate is 0.6-1.0ml/min, the column temperature is 22-28 ℃, and the sample injection amount is 1-20 mu l; and/or, use a Ultimate XB-C18 column of size 4.6x250mm,5 μm.
Further, the construction method also comprises the steps of preparing a reference substance solution by adopting at least one of Wang Baige glycoside A, wang Baige glycoside H and 2-acetyl Wang Baige glycoside A reference substance and detecting the reference substance solution by using a high performance liquid chromatography in any one of the construction methods to obtain a reference substance reference map; optionally, the reference substance solution adopts 30-70vt% methanol water solution; optionally, the concentration of the control solution is 10-100 mug/mL.
Further, the lily medicinal materials with different origins and the preparation thereof are as follows: lily medicinal material of Paeonia ostii and preparation thereof, lily medicinal material of Paeonia ostii and preparation thereof, or lily medicinal material of Paeonia ostii and preparation thereof.
In the invention, the term lily medicinal material of a proscenium substrate and a preparation thereof, lily (proscenium substrate) and a preparation thereof refer to lily medicinal materials, decoction pieces or preparations from proscenium substrate. The term lily medicinal material of lily-based raw material and its preparation, lily (lily) and its preparation refer to lily medicinal material, decoction piece or preparation of lily source. The term lily medicinal material of the fine leaf lily base raw material and preparation thereof, lily (fine leaf lily) and preparation thereof refer to lily medicinal material, decoction pieces or preparation from the fine leaf lily. The preparation can be, but not limited to, powder, formulated granules and other conventional dosage forms. Lily (lilium tenuifolium) is abbreviated as lilium tenuifolium. Lily (roll pill) is called roll pill for short.
Further, the characteristic patterns of the lily medicinal material and the preparation thereof have 9 characteristic peaks, the peak corresponding to the Wang Baige glycoside A reference substance peak is an S peak, the relative retention time of each characteristic peak and the S peak is within the range of +/-10% of a specified value, and the specified value is: 0.50 (peak 1), 0.62 (peak 2), 0.91 (peak 3), 1.02 (peak 5), 1.04 (peak 6), 1.30 (peak 7), 1.43 (peak 8), 1.46 (peak 9);
the lily medicinal material of the original rolling dansyl and the preparation thereof have 10 characteristic peaks, the peak corresponding to the reference peak of the Wang Baige glycoside A reference substance is an S peak, the relative retention time of each characteristic peak and the S peak is within the range of +/-10% of a specified value, and the specified value is: 0.50 (peak 1), 0.62 (peak 2), 0.91 (peak 3), 1.02 (peak 5), 1.04 (peak 6), 1.30 (peak 7), 1.43 (peak 8), 1.46 (peak 9); a chromatographic peak (peak 10) appears at the corresponding position of the chromatographic peak of the 2-acetyl Wang Baige glycoside A (peak 10) reference substance;
the lily medicinal material of the fine leaf lily base stock and the preparation thereof have 11 characteristic peaks, the peak corresponding to the Wang Baige glycoside A reference substance peak is an S peak, the relative retention time of each characteristic peak and the S peak is within the range of +/-10% of a specified value, and the specified value is: 0.50 (Peak 1), 0.62 (Peak 2), 0.91 (Peak 3), 1.02 (Peak 5), 1.04 (Peak 6), 1.30 (Peak 7), 1.43 (Peak 8), 1.46 (Peak 9), 1.49 (Peak 11), 1.50 (Peak 12).
The peak corresponding to the Wang Baige glycoside A reference peak in the invention is a characteristic peak with a retention time of less than 5%, 3% or 1% of the peak of the Wang Baige glycoside A reference peak and/or a characteristic peak with a coincidence degree of not less than 50% of the peak of the Wang Baige glycoside A reference peak.
Further, the chromatographic peak appearing at the corresponding position of the chromatographic peak of the 2-acetyl Wang Baige glycoside A reference substance is the original rolling dansyl, and the colorless chromatographic peak is the original lily or fine leaf lily.
The method also comprises the step of adopting fingerprint similarity evaluation software '2012 edition of traditional Chinese medicine chromatographic fingerprint similarity evaluation system' compiled by the pharmacopoeia committee to generate a comparison characteristic spectrum, and obtaining the lily medicinal material HPLC characteristic spectrum with 9-11 chromatographic peaks through the identification and the assignment of the characteristic peaks.
The invention also provides a method for identifying lily medicinal materials with different origins and preparations thereof, which comprises the step of obtaining the characteristic spectrum of lily products to be identified according to any one of the construction methods.
If the lily product to be identified has a chromatographic peak at the corresponding position of the chromatographic peak of the 2-acetyl Wang Baige glycoside A reference substance, the lily product to be identified is a rolling stock, if the lily product to be identified has no chromatographic peak at the corresponding position of the chromatographic peak of the 2-acetyl Wang Baige glycoside A reference substance and the relative peak area of the peak 9 and the peak 4 is smaller than 0.23, the lily product is a fine leaf lily stock, and if the lily product has no color spectrum and the relative peak area of the peak 9 and the peak 4 is larger than or equal to 0.23, the lily product is a lily stock.
The technical scheme of the invention has the following advantages:
1. according to the construction method of the characteristic spectrum of lily medicinal materials with different basic origins and preparations thereof, through investigation of chromatographic conditions, the invention discovers that octadecylsilane chemically bonded silica is used as a filler, a mobile phase comprises acetonitrile and 0.05-0.2vt% of phosphoric acid aqueous solution, gradient elution is carried out, and a gradient elution program comprises: 0 to 10 minutes to 15 minutes to 23 minutes to 30 minutes to 35 minutes, wherein the volume percentage of acetonitrile in the mobile phase is 6 to 12 to 18 to 40 to 70 percent, the complete separation of 9 to 11 common characteristic peaks including Wang Baige glycoside A and Wang Baige glycoside H, including 2-acetyl Wang Baige glycoside A and the like for the original rolling dansyl is realized, the peak shape is good, the base line is stable, the detection time is short, the characteristic patterns of lily medicinal materials with different base sources and preparations thereof are constructed, the chemical component characteristics of lily with different base sources are fully displayed, the basis is provided for identifying and distinguishing lily medicinal materials with different base sources and preparations thereof, and the method is stable, high in precision, good in stability, strong in characteristic and rich in chromatographic information.
2. The identification method of the lily medicinal materials and the preparations thereof with different base stocks can be used for identifying and distinguishing lily medicinal materials and the preparations thereof with the prosoma lilium, and lily medicinal materials and the preparations thereof with the prosoma lilium gracilis, the result displayed by the atlas is easy to judge by the mode of positioning or combining the relative peak area with the 2-acetyl Wang Baige glycoside A reference substance, the identification method can be directly judged without calculating the similarity with the reference characteristic atlas, the problem that the prosoma lilium and the prosoma lilium are difficult to judge in the prior art is solved, and a rapid and reliable detection method is provided for the interplanar identification of the lily medicinal materials.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the description of the embodiments or the prior art will be briefly described, and it is obvious that the drawings in the description below are some embodiments of the present invention, and other drawings can be obtained according to the drawings without inventive effort for a person skilled in the art.
FIG. 1 is a characteristic spectrum of a lily (lily) herb sample in example 1;
FIG. 2 is a characteristic spectrum of a sample of lily (lily) pieces in example 2;
FIG. 3 is a characteristic spectrum of a standard lily (lily) decoction lyophilized powder sample of example 3;
FIG. 4 is a characteristic spectrum of 18 batches of lily (lily) in example 4;
FIG. 5 is a comparison characteristic map of lily (lily) medicinal material in example 4;
FIG. 6 is a characteristic spectrum of 18 batches of lily (lily) pieces of example 5;
FIG. 7 shows the comparative characteristic patterns of examples 4-6, wherein the comparative characteristic patterns of the medicinal materials, the comparative characteristic patterns of the decoction pieces and the comparative characteristic patterns of the standard decoction are sequentially shown from bottom to top;
fig. 8 is a characteristic spectrum of lily medicinal materials of different origins in example 7, S1: 2-acetyl Wang Baige glycoside A control solution; s2: lily (lily) control medicinal material; S3-S8 are 6 batches of lily (red roll) medicinal materials respectively, and S9: lily (roll pill) control medicinal material;
Fig. 9 is a characteristic spectrum of lily medicinal materials of different origins in example 7, S1: lily (lily) control medicinal material; S2-S3 are two batches of lily (lilium tenuifolium) medicinal materials;
FIG. 10 is a characteristic spectrum of lily standard decoction freeze-dried powder of different primordia in example 8, S1:2-acetyl Wang Baige glycoside A; s2: standard lily (lily) decoction freeze-dried powder; S3-S8 are 6 batches of lily (red roll) standard decoction freeze-dried powder respectively; s9: lily (roll pill) control medicinal material;
FIG. 11 is a characteristic diagram of lily standard decoction lyophilized powder of different origins in example 8, S1, lily (Bulbus Lilii) standard decoction lyophilized powder; S2-S3 are two batches of standard decoction freeze-dried powder of lily (fine leaf lily);
FIG. 12 is a characteristic spectrum obtained by the Agilent SB AQ column in example 9;
FIG. 13 is a characteristic spectrum obtained by YMC-Pack ODS-AQ column in example 9;
FIG. 14 is a characteristic spectrum obtained by Ultimate XB-C18 column in example 9;
FIG. 15 is a characteristic spectrum obtained in the gradient elution procedure 1 of example 9;
FIG. 16 is a characteristic spectrum obtained in the gradient elution procedure 2 of example 9;
FIG. 17 is a characteristic spectrum obtained in the gradient elution procedure 3 of example 9;
FIG. 18 is a characteristic spectrum obtained in the gradient elution procedure 4 of example 9;
FIG. 19 is a characteristic spectrum obtained in the gradient elution procedure 5 of example 9;
FIG. 20 is a characteristic spectrum obtained in the gradient elution procedure 6 of example 9;
FIG. 21 is a characteristic spectrum obtained in the gradient elution procedure 7 of example 9;
FIG. 22 is a characteristic spectrum of a lily medicinal material, an ultraviolet absorption spectrum of each characteristic peak, and an ultraviolet absorption spectrum of Wang Baige glycoside H (left side) and Wang Baige glycoside A (right side) in a sample, wherein the characteristic spectrum of the lily medicinal material RK21050703 is sequentially shown from top to bottom;
FIG. 23 shows the control chromatogram and the ultraviolet absorption spectrum, from top to bottom, the control chromatogram of lilac glycoside H, the control chromatogram of Wang Baige glycoside A, and the ultraviolet absorption spectrum of control Wang Baige glycoside H (left side) and Wang Baige glycoside A (right side).
Detailed Description
The following examples are provided for a better understanding of the present invention and are not limited to the preferred embodiments described herein, but are not intended to limit the scope of the invention, any product which is the same or similar to the present invention, whether in light of the present teachings or in combination with other prior art features, falls within the scope of the present invention.
The specific experimental procedures or conditions are not noted in the examples and may be followed by the operations or conditions of conventional experimental procedures described in the literature in this field. The reagents or apparatus used were conventional reagent products commercially available without the manufacturer's knowledge.
Experimental instruments and reagents used in the examples and experimental examples of the present invention are as follows.
High performance liquid chromatograph 1: waters e2695, including a quaternary pump of G7104A, a 132 bit autosampler, a column oven of G7116B, a diode array detector.
High performance liquid chromatograph 2: agilent 1260 comprising G7111 aqut Pump VL; G7129A 1260Vialsamplar; G7114A1260VWD.
High performance liquid chromatograph 3: thermo U3000 chromatographic systems, including quaternary solvent manager (pμmp), autosampler (Autosampler), original-in-place inlet chromatographic column incubator (Col μmn component), diode array ultraviolet Detector (Detector), chromeleon chromatographic management system.
Electronic analytical balance: one ten thousandth balance MS205DU (Metrele), one ten thousandth balance FA2004 (Shunhai constant level), one-day balance LD610-2 (Shenyang Dragon electron).
Ultrasonic cleaning machine: BILON model 30-720, shanghai Bilang instruments Co., ltd.
Chromatographic column 1: agilent SB AQ,4.6X250mm 5 μm.
Chromatographic column 2: YMC-Pack ODS-AQ,4.6X250mm 5 μm.
Chromatographic column 3: hypersil GOLD AQ C18,4.6X250mm 5 μm.
Reagent: acetonitrile is chromatographic pure, and water is ultrapure water; other reagents such as phosphoric acid are analytically pure.
Wang Baige glycoside A control, lot number: 114420-66-5 from Chengdu biosciences Inc.
Lily control drug, lot number: 121100-201906, available from China food and drug verification institute.
Wang Baige glycoside H control, lot number: PS012362 has purity not less than 98.0%, and is available from Chengdu Pusi Biotech Co., ltd.
The standard decoction freeze-dried powder can be prepared by adopting the conventional technical means in the field, and the standard decoction freeze-dried powder of lily (lily), the standard decoction freeze-dried powder of lily (lilium schoendole) and the standard decoction freeze-dried powder of lily (fine leaf lily) are respectively prepared by adopting lily (lily) medicinal materials, lily (lilium schoendole) medicinal materials and lily (fine leaf lily) medicinal materials according to the following steps.
Taking lily medicinal materials, and processing (processing technique: lily, removing impurities) according to the relevant regulations of the 2020 edition of Chinese pharmacopoeia to obtain lily decoction pieces meeting the requirements. About 150g of lily decoction pieces are taken, put into a sand tank, added with water with the weight being 10 times that of the lily decoction pieces, soaked for 30 minutes, boiled with strong fire, decocted with slow fire for 30 minutes, filtered with 200-mesh filter cloth while the decoction pieces are hot, added with water with the weight being 8 times that of the lily decoction pieces, decocted with slow fire for 25 minutes after the decoction pieces are boiled with strong fire, filtered with 200-mesh filter cloth while the decoction pieces are hot, and the filtrates are combined; concentrating the filtrate under reduced pressure (65deg.C) to obtain fluid extract with relative density of about 1.02; freeze drying, taking out, mixing, packaging, and sealing to obtain Bulbus Lilii standard decoction lyophilized powder.
Example 1
The embodiment provides a construction method of lily medicinal material characteristic patterns, which comprises the following steps:
(1) Preparation of test solution: taking lily (lily) medicinal material as a test sample, taking 1g of the test sample, placing the test sample into a conical bottle with a plug, adding 50ml of water, heating and refluxing for 30 minutes, filtering, evaporating filtrate to dryness, performing ultrasonic treatment (250W, 40 KHz) on residues by using 20ml of 50% methanol for 30 minutes, cooling, uniformly mixing, filtering, and taking the subsequent filtrate as a test sample solution.
(2) High performance liquid chromatography: precisely sucking 10 μl of the sample solution, injecting into a liquid chromatograph, and measuring under the following chromatographic conditions: octadecylsilane chemically bonded silica is used as filler (column length of 250mm, inner diameter of 4.6mm, and particle diameter of 5 μm), and the chromatographic column is XU Ultimate XB-C18,4.6mm×250mm,5 μm column; acetonitrile is taken as a mobile phase A,0.1% phosphoric acid is taken as a mobile phase B, and gradient elution is carried out according to the specifications in the following table; the flow rate is 0.8ml per minute; the column temperature is 25 ℃; the detection wavelength is 205nm,15 min tangential wavelength is 310nm, and theoretical plate number is Wang Baige glycoside A to be no less than 3000.
The results are shown in the following table.
TABLE 1 Peak results of Lily herbs
The results are shown in the table and in fig. 1, the characteristic spectrum of lily (lily) medicinal material has 9 characteristic peaks, peak 4 is taken as a reference peak, and the relative retention time of peaks 1-3 and 5-9 and peak 4 are respectively as follows: 0.50, 0.62, 0.91, 1.02, 1.03, 1.30, 1.44 and 1.47.
Example 2
The embodiment provides a construction method of lily decoction piece characteristic patterns, which comprises the following steps:
(1) Preparation of test solution: taking lily medicinal materials, removing impurities to obtain lily (lily) decoction pieces, taking lily (lily) decoction pieces as a test sample, taking 1g of the test sample, placing the test sample into a conical flask with a plug, adding 50ml of water, heating and refluxing for 30 minutes, filtering, evaporating filtrate to dryness, performing ultrasonic treatment (250W, 40 KHz) on residues with 20ml of 50% methanol for 30 minutes, cooling, uniformly mixing, filtering, and taking the subsequent filtrate as a test sample solution.
(2) Preparation of a control solution: taking a proper amount of Wang Baige glycoside H, wang Baige glycoside A and 2-acetyl Wang Baige glycoside A reference substances, precisely weighing, adding 50% methanol to prepare a mixed solution containing Wang Baige glycoside H40 mug, wang Baige glycoside 40 mug and 2-acetyl Wang Baige glycoside 40 mug per 1ml, shaking uniformly, and taking the mixed solution as a reference substance solution;
(3) High performance liquid chromatography: precisely sucking 10 μl of the sample solution and reference solution, injecting into a liquid chromatograph, and measuring under the following chromatographic conditions: octadecylsilane chemically bonded silica is used as filler (column length 250mm, inner diameter 4.6mm, particle diameter 5 μm); acetonitrile is taken as a mobile phase A,0.1% phosphoric acid is taken as a mobile phase B, and gradient elution is carried out according to the specifications in the following table; the flow rate is 0.8ml per minute; the column temperature is 25 ℃; the detection wavelength is 205nm,15 min tangential wavelength is 310nm, and theoretical plate number is Wang Baige glycoside A to be no less than 3000.
The results are shown in the following table.
TABLE 2 Peak results of Lily decoction pieces
The results are shown in the table and fig. 2, the characteristic spectrum of lily (lily) decoction pieces has 9 characteristic peaks, the peak 4 is taken as a reference peak, and the relative retention time of the peaks 1-3 and 5-9 and 4 is respectively as follows: 0.50, 0.62, 0.91, 1.02, 1.03, 1.30, 1.44 and 1.46. Wherein, the peak 3 and the peak 4 are a lilac glycoside H peak and a Wang Baige glycoside A peak, and no 2-acetyl Wang Baige glycoside A peak.
Example 3
The embodiment provides a construction method of a lily standard decoction freeze-dried powder characteristic spectrum, which comprises the following steps:
(1) Preparation of test solution: preparing test solution, namely taking lily standard decoction freeze-dried powder as a test sample, wherein the batch number is 21051801 (corresponding medicinal material is RK 21050703), taking 0.2g of the test sample, precisely weighing, placing in a conical bottle with a plug, adding 20ml of 50% methanol, weighing, performing ultrasonic treatment (250W, 40 KHz) for 30 minutes, cooling, supplementing the weight of loss with 50% methanol, uniformly mixing, filtering, and taking the subsequent filtrate.
(2) Preparation of a control solution: as in example 2.
(3) High performance liquid chromatography: as in example 2.
TABLE 3 Peak results for Lily Standard decoction lyophilized powder
The results are shown in the table and in fig. 3, the characteristic spectrum of lily (lily) decoction pieces has 9 characteristic peaks in total, and peak 4 is taken as a reference peak, and the relative retention time of peaks 1-3 and peaks 5-9 and peak 4 are respectively as follows: 0.45, 0.58, 0.90, 1.02, 1.03, 1.25, 1.46 and 1.50. Wherein, the peak 3 and the peak 4 are a lilac glycoside H peak and a Wang Baige glycoside A peak, and no 2-acetyl Wang Baige glycoside A peak.
Example 4
The embodiment provides a construction method of lily medicinal material characteristic patterns, which comprises the following steps:
(1) Preparation of test solution: taking 18 batches of lily (lily) medicinal materials as test substances, and preparing a test substance solution according to the method of the example 1;
(2) Preparation of reference solution: about 1g of lily (lily) reference medicine is taken, put into a conical flask with a plug, added with 50ml of water, heated and refluxed for 30 minutes, filtered, evaporated to dryness, added with 20ml of 50% methanol in residues, subjected to ultrasonic treatment (250W, 40 KHz) for 30 minutes, cooled, shaken uniformly, filtered, and taken as a reference substance solution of the reference medicine. Taking a proper amount of Wang Baige glycoside H and Wang Baige glycoside A reference substances, precisely weighing, and adding 50% methanol to prepare a solution containing 40 mug of each reference substance per 1 ml.
(3) High performance liquid chromatography: 10. Mu.l of the sample solution was precisely aspirated, and the sample solution was injected into a liquid chromatograph to measure the sample solution, and the chromatographic conditions were the same as those in example 1.
And (3) generating a control characteristic spectrum by adopting characteristic spectrums of 18 samples (shown in fig. 4) and adopting fingerprint spectrum similarity evaluation software 'a traditional Chinese medicine chromatographic fingerprint spectrum similarity evaluation system 2012 edition' compiled by pharmacopoeia committee, wherein the total number of the obtained lily medicinal material HPLC characteristic spectrums is 9 by identifying and identifying characteristic peaks, and the chromatographic peak numbers 1-9 are arranged according to the sequence of the chromatographic peaks (shown in fig. 5). And determining that the peak 3 is the wang lily glycoside H and the peak 4 is the wang lily glycoside A according to the peak identification and the peak selection result of the characteristic peak in the lily medicinal material, wherein the peak of the reference substance has larger peak response and better stability, and the peak is an index component of content measurement control, and taking Wang Baige glycoside A as an S peak of the characteristic map. The relative retention time of each characteristic peak was calculated with Wang Baige glycoside a as the S peak. And the relative peak areas of the peaks and S peaks were calculated.
According to the reference medicinal material, the corresponding peaks of 18 batches of medicinal materials and the reference substance (Wang Baige glycoside A) are S peaks, the relative retention time of each characteristic peak and S peak is calculated to be within the range of +/-10% of the mean value, so that a lily characteristic spectrum standard is established, namely 9 characteristic peaks should be presented in a sample characteristic spectrum and correspond to the relative retention time of 9 characteristic peaks in the chromatogram of the reference substance of the reference medicinal material, the corresponding peak of Wang Baige glycoside A reference substance is taken as the S peak, the relative retention time of peaks 1-3, peaks 5-9 and S peaks is calculated, the relative retention time of each characteristic peak is specified to be within the range of +/-10% of a specified value, and the specified value is: 0.50 (Peak 1), 0.62 (Peak 2), 0.91 (Peak 3), 1.02 (Peak 5), 1.04 (Peak 6), 1.30 (Peak 7), 1.43 (Peak 8), 1.46 (Peak 9).
The peak areas of the 18 batches of medicinal material characteristic maps are large in difference, the peak area of each peak in the batches of medicinal materials is 0.254-0.801 with respect to the peak 1 of Wang Baige glycoside A, the peak 2 is 0.348-0.980, the peak 3 is 0.109-0.158, the peak 5 is 0.093-0.275, the peak 6 is 0.036-0.208, the peak 7 is 0.022-0.096, the peak 8 is 0.158-0.490 and the peak 9 is 0.331-0.525; and combining characteristic spectrum identification results, wherein the peak 3 is the lilac glycoside H, the peak 4 is the lilac glycoside A, and combining characteristic spectrum conditions of different base sources, the relative peak areas of the peak 9 and the peak 4 are temporarily determined.
TABLE 4 characteristic peak relative retention times
TABLE 5 characteristic peak to peak area
Example 5
The embodiment provides a construction method of lily decoction piece characteristic patterns, which comprises the following steps:
(1) Preparation of test solution: taking 18 batches of lily (lily) decoction pieces as test samples, and preparing a test sample solution according to the method of the example 2;
(2) High performance liquid chromatography: 10. Mu.l of the sample solution was precisely aspirated, and the sample solution was injected into a liquid chromatograph to measure the sample solution, and the chromatographic conditions were the same as those in example 2.
And (3) generating a control characteristic spectrum by adopting characteristic spectrums of 18 samples (shown in fig. 6) and adopting fingerprint spectrum similarity evaluation software 'a traditional Chinese medicine chromatographic fingerprint spectrum similarity evaluation system 2012 edition' compiled by pharmacopoeia committee, wherein the total number of the obtained lily medicinal material HPLC characteristic spectrums is 9 by identifying and identifying characteristic peaks, and the chromatographic peak numbers 1-9 are arranged according to the sequence of the chromatographic peaks (shown in fig. 7). The results are shown in the following table, the corresponding peak of Wang Baige glycoside A reference substance peaks is taken as an S peak, the relative retention time of peaks 1-3, peaks 5-9 and the S peak in 18 batches of lily (lily) decoction piece characteristic maps is calculated to be within +/-10% of a specified value, and the specified value is: 0.50 (Peak 1), 0.62 (Peak 2), 0.91 (Peak 3), 1.02 (Peak 5), 1.04 (Peak 6), 1.30 (Peak 7), 1.43 (Peak 8), 1.46 (Peak 9) are all satisfactory. And the relative peak area of peak 9 and peak 4 is not less than 0.23.
TABLE 6 peak to peak retention times of decoction pieces
TABLE 7 Peak to Peak area characteristic of decoction pieces
Example 6
The embodiment provides a construction method of a lily standard decoction freeze-dried powder characteristic spectrum, which comprises the following steps:
(1) Preparation of test solution: taking 18 batches of lily (lily) medicinal materials to obtain standard decoction freeze-dried powder as a test sample, and preparing a test sample solution according to the method of the example 3;
(2) High performance liquid chromatography: 10. Mu.l of the sample solution was precisely aspirated, and the sample solution was injected into a liquid chromatograph to measure the sample solution, and the chromatographic conditions were the same as those in example 3.
The obtained characteristic spectrum of 18 samples adopts fingerprint spectrum similarity evaluation software programmed by pharmacopoeia committee, namely 2012 edition of traditional Chinese medicine chromatographic fingerprint spectrum similarity evaluation system, a control characteristic spectrum is generated, 9 chromatographic peaks are shared by the obtained lily medicinal material HPLC characteristic spectrum through identification and assignment of characteristic peaks, chromatographic peak numbers 1-9 are arranged according to the sequence of the chromatographic peaks (as shown in figure 7), the corresponding peak of Wang Baige glycoside A reference substance peak is taken as an S peak, and the relative retention time of peaks 1-3, peaks 5-9 and the S peak in 18 batches of lily (lily) standard decoction freeze-dried powder characteristic spectrum is within a range of +/-10% of a specified value, wherein the specified value is: 0.50 (Peak 1), 0.62 (Peak 2), 0.91 (Peak 3), 1.02 (Peak 5), 1.04 (Peak 6), 1.30 (Peak 7), 1.43 (Peak 8), 1.46 (Peak 9) are all satisfactory. And the relative peak area of peak 9 and peak 4 is not less than 0.23.
TABLE 8 results of characteristic peaks of standard decoction (lyophilized powder) of 18 batches of lily (Bulbus Lilii) versus retention time
TABLE 9 results of characteristic peak to peak area of standard decoction (lyophilized powder) of 18 batches of lily (Bulbus Lilii)
Example 7
The embodiment provides a method for identifying lily medicinal materials with different origins, which comprises the following steps:
(1) Preparation of test solution: preparing test solution by taking 6 batches of lily (Paeonia ostii) medicinal materials and 2 batches of lily (Lilium tenuifolium) medicinal materials as test substances respectively, taking 1g of the test substances, placing in a conical flask with a plug, adding 50ml of water, heating and refluxing for 30 minutes, filtering, evaporating filtrate to dryness, performing ultrasonic treatment (250W, 40 KHz) on residues by using 20ml of 50% methanol for 30 minutes, cooling, uniformly mixing, filtering, and taking the subsequent filtrate as the test substance solution.
(2) Preparation of reference solution: about 1g of lily (lily) control medicinal material (provided by Chinese inspection hospital) and lily (lily) control medicinal material (provided by Chinese inspection hospital) are respectively taken, put into a conical flask with a plug, added with 50ml of water, heated and refluxed for 30 minutes, filtered, evaporated to dryness, added with 20ml of 50% methanol into residues, subjected to ultrasonic treatment (250W, 40 KHz) for 30 minutes, cooled, shaken uniformly, filtered, and taken as a reference solution of the control medicinal material. Taking 2-acetyl Wang Baige glycoside A reference substance with proper amount, precisely weighing, adding 50% methanol to prepare solution containing 40 μg 2-acetyl Wang Baige glycoside A per 1 ml.
(3) High performance liquid chromatography: precisely sucking 10 μl of the sample solution and the reference solution, respectively, injecting into a liquid chromatograph, and measuring under the following chromatographic conditions: octadecylsilane chemically bonded silica is used as filler (column length 250mm, inner diameter 4.6mm, particle diameter 5 μm); acetonitrile is taken as a mobile phase A,0.1% phosphoric acid is taken as a mobile phase B, and gradient elution is carried out according to the specifications in the following table; the flow rate is 0.8ml per minute; the column temperature is 25 ℃; the detection wavelength is 205nm,15 min tangential wavelength is 310nm, and theoretical plate number is Wang Baige glycoside A to be no less than 3000.
Compared with lily (lily) medicinal materials, the characteristic spectrum relative peak area of lily (prosopam) medicinal materials is compared with that of 18 batches of lily (lily) medicinal materials, and the results are shown in the following table.
Table 10 Lily bulb extract characteristic spectrum relative peak area
Compared with lily (lily) medicinal materials, the characteristic spectrum relative peak area of lily (fine leaf lily) medicinal materials is compared with that of 18 batches of lily (lily) medicinal materials, and the results are shown in the following table.
Table 11 Lily bulb extract characteristic spectrum relative peak area
The results are shown in fig. 8 and 9 and the table above, compared with the lily (lily) medicinal material, the peak 9 characteristic peak response value in the lily (lilium schoendole) medicinal material and the lily (lilium tenuifolium) medicinal material characteristic spectrum is extremely low and is obviously lower than that of the lily (lily) medicinal material, for example, when the relative peak area of the peak 9 and the peak 4 is smaller than 0.23, the lily (lilium schoendole) medicinal material is obtained; when the relative peak area of the peak 9 and the peak 4 is more than or equal to 0.23, the medicinal material is lily (lily).
In addition, 2-acetyl Wang Baige glycoside A chromatographic peaks do not exist in the characteristic maps of lily (lily) medicinal materials and lily (lilium tenuifolium) medicinal materials, but peaks 11 and 12 are obviously exist in the lilium tenuifolium (lily) medicinal materials, and no lily (lily tenuifolium) medicinal materials and lily (lilium tenuifolium) medicinal materials exist.
The lily medicinal material of the original rolling dansyl has 10 characteristic peaks, the peak corresponding to the Wang Baige glycoside A reference substance peak is an S peak, the relative retention time of each characteristic peak and the S peak is within the range of +/-10% of a specified value, and the specified value is: 0.50 (peak 1), 0.62 (peak 2), 0.91 (peak 3), 1.02 (peak 5), 1.04 (peak 6), 1.30 (peak 7), 1.43 (peak 8), 1.46 (peak 9); a chromatographic peak appears at the corresponding position of the chromatographic peak of the 2-acetyl Wang Baige glycoside A (peak 10) reference substance;
the lily medicinal material of the fine leaf lily base stock has 11 characteristic peaks, the peak corresponding to the Wang Baige glycoside A reference substance peak is an S peak, the relative retention time of each characteristic peak and the S peak is within the range of +/-10% of a specified value, and the specified value is: 0.50 (Peak 1), 0.62 (Peak 2), 0.91 (Peak 3), 1.02 (Peak 5), 1.04 (Peak 6), 1.30 (Peak 7), 1.43 (Peak 8), 1.46 (Peak 9), 1.49 (Peak 11), 1.50 (Peak 12).
Example 8
The embodiment provides a method for identifying lily standard decoction freeze-dried powder with different primordia, which comprises the following steps:
(1) Preparation of test solution: taking 6 batches of standard decoction freeze-dried powder of lily (lilium tenuifolium) and 2 batches of standard decoction freeze-dried powder of lily (lilium tenuifolium) as test products respectively, taking about 0.2g of test product powder, precisely weighing, placing into a conical bottle with a plug, precisely adding 20ml of 50% methanol, weighing, performing ultrasonic treatment (250W, 40 KHz) for 30 minutes, cooling, weighing again, supplementing the lost weight with 50% methanol, uniformly mixing, filtering, and taking the subsequent filtrate.
(2) Preparation of reference solution: about 1g of lily (volume pill) reference medicinal material (provided by Chinese inspection hospital) is taken, put into a conical flask with a plug, added with 50ml of water, heated and refluxed for 30 minutes, filtered, evaporated to dryness, and the residue is added with 20ml of 50% methanol, treated by ultrasonic (250W, 40 KHz) for 30 minutes, cooled, shaken uniformly, filtered, and the subsequent filtrate is taken as reference substance solution of the reference medicinal material. Taking 2-acetyl Wang Baige glycoside A reference substance with proper amount, precisely weighing, adding 50% methanol to prepare solution containing 40 μg 2-acetyl Wang Baige glycoside A per 1 ml.
(3) High performance liquid chromatography: same as in example 7.
As shown in fig. 10 and 11 and the following table, 2-acetyl Wang Baige glycoside a chromatographic peak exists in the characteristic spectrum of the standard decoction freeze-dried powder of lily (lilium) and fine leaf lily, 2-acetyl Wang Baige glycoside a chromatographic peak does not exist in the standard decoction freeze-dried powder of lily (lilium) and fine leaf lily, 2-acetyl Wang Baige glycoside a chromatographic peak does not exist in the characteristic spectrum of the fine leaf lily freeze-dried powder, but chromatographic peak 11 and chromatographic peak 12 are obvious.
The characteristic spectrum relative peak area of the standard lily (lilium) decoction freeze-dried powder is compared with that of 18 batches of standard lily (lilium) decoction freeze-dried powder, and the results are shown in the following table.
Table 12 characteristic spectrum of Standard decoction of Papaver and Lily relative peak area
The characteristic spectrum relative peak area of the standard lily (lilium tenuifolium) decoction lyophilized powder is compared with that of 18 batches of standard lily (lilium tenuifolium) decoction lyophilized powder, and the results are shown in the following table.
Table 13 Standard decoction feature patterns of Lily tenuifolia versus Peak area
The results are shown in fig. 10 and 11 and the table above, wherein the relative peak area of peak 8 in 18 batches of lily (lily) standard decoction lyophilized powder ranges from 0.19 to 0.28, the mean value is 0.24, and the relative peak area of peak 9 ranges from 0.315 to 0.44, and the mean value is 0.37. Peak 8 and Peak 9 integration difficulties in standard lily (Paeonia suffruticosa) freeze-dried powder with a peak area range of 0.00-0.01 and a peak 9 relative to a peak area range of 0.04-0.07. Considering comprehensively that the relative peak area of the standard lily (lily) decoction freeze-dried powder peak 9 is regulated, the standard lily (lily) decoction freeze-dried powder is recommended to be regulated according to-30% of the lower limit of a plurality of batches of practical measurement ranges (peak 9:0.315 x 0.7= 0.2205, and the standard lily (lily) decoction freeze-dried powder is regulated to be 0.23 or more when the relative peak area of the peak 9 and the peak 4 in the standard lily decoction freeze-dried powder to be identified is 0.23 or more, and the standard lily (lily) decoction freeze-dried powder is regulated to be lily (prosoma) or lily (fine-leaf lily) standard decoction freeze-dried powder when the relative peak area of the peak 9 and the peak 4 in the standard lily decoction freeze-dried powder to be identified is less than 0.23, and the standard lily (prosoma) decoction freeze-dried powder is regulated to be lily (prosoma) when the 2-acetyl Wang Baige glycoside A chromatographic peak exists.
The lily standard decoction freeze-dried powder of the original rolling dansyl has 10 characteristic peaks, the peak corresponding to the Wang Baige glycoside A reference substance peak is an S peak, the relative retention time of each characteristic peak and the S peak is within the range of +/-10% of a specified value, and the specified value is: 0.50 (peak 1), 0.62 (peak 2), 0.91 (peak 3), 1.02 (peak 5), 1.04 (peak 6), 1.30 (peak 7), 1.43 (peak 8), 1.46 (peak 9); a chromatographic peak (peak 10) appears at the corresponding position of the chromatographic peak of the 2-acetyl Wang Baige glycoside A (peak 10) reference substance;
the lily standard decoction freeze-dried powder of the fine leaf lily base source has 11 characteristic peaks, wherein the peak corresponding to the Wang Baige glycoside A reference substance peak is an S peak, the relative retention time of each characteristic peak and the S peak is within the range of +/-10% of a specified value, and the specified value is: 0.50 (Peak 1), 0.62 (Peak 2), 0.91 (Peak 3), 1.02 (Peak 5), 1.04 (Peak 6), 1.30 (Peak 7), 1.43 (Peak 8), 1.46 (Peak 9), 1.49 (Peak 11), 1.50 (Peak 12).
Example 9
1. Instrument:
high performance liquid chromatograph 1: waters e2695, including a quaternary pump of G7104A, a 132 bit autosampler, a column oven of G7116B, a diode array detector.
High performance liquid chromatograph 2: agilent 1260 comprising G7111A Quat Pump VL; G7129A 1260Vialsamplar; G7114A 1260VWD.
Electronic analytical balance: METTER TOLEDO (METHOD. Switzerland) ME36S, XS, XS205, XSE205 (parts per million); SK5200H Shanghai Kogyo ultrasonic instruments Co.
Chromatographic column 1: agilent SB AQ,4.6x250mm 5 μm;
chromatographic column 2: YMC-Pack ODS-AQ,4.6x250mm 5 μm;
chromatographic column 3: ultimate XB-C18,4.6x250mm 5 μm;
2. reagent and reagent:
reagent: acetonitrile is chromatographic pure, and water is ultrapure water; other reagents such as phosphoric acid are analytically pure.
Wang Baige glycoside A reference (lot number 114420-66-5, available from Chengdu Posi Biotechnology Co., ltd.)
Lily control medicinal material (lot number 121100-201906, purchased from China food and drug inspection institute).
The preparation method of the sample solution comprises the following steps: same as in example 7.
3. Investigation experiment of chromatographic conditions
(1) Screening experiments on chromatography columns
The sample solutions were tested using the following three different chromatographic columns.
Chromatographic condition 1: agilent SB AQ,4.6x250mm 5 μm chromatographic column, acetonitrile as mobile phase A,0.1% phosphoric acid as mobile phase B, eluting according to gradient 1; the flow rate is 0.8ml per minute; the column temperature is 25 ℃; the detection wavelength is 205nm, and the 15 minute wavelength is 310nm.
Chromatographic condition 2: YMC-Pack ODS-AQ,4.6x250mm 5 μm chromatographic column, acetonitrile as mobile phase A,0.1% phosphoric acid as mobile phase B, eluting according to gradient 1; the flow rate is 0.8ml per minute; the column temperature is 25 ℃; the detection wavelength is 205nm, and the 15 minute wavelength is 310nm. Chromatographic condition 3: ultimate XB-C18,4.6x250mm 5 μm chromatographic column, eluting with acetonitrile as mobile phase A and 0.1% phosphoric acid as mobile phase B according to gradient 1; the flow rate is 0.8ml per minute; the column temperature is 25 ℃; the detection wavelength is 205nm, and the 20 minute wavelength is 310nm.
The test results are shown in figures 12-14, the characteristic spectrum peak separation of the Ultimate XB-C18 chromatographic column is better than that of YMC-Pack ODS-AQ and YMC-Pack ODS-AQ chromatographic columns, and the target peaks are more, so that the Ultimate XB-C18.6x250mm 5 μm chromatographic column is selected for further investigation.
(2) Gradient optimization
Taking the sample solution, respectively testing under the following gradient elution procedures, eluting by taking acetonitrile as a mobile phase A and 0.1% phosphoric acid as a mobile phase B according to a gradient elution procedure 1; the flow rate is 0.8ml per minute; the column temperature is 25 ℃; the detection wavelength is 205nm, and the gradient is 20 minutes tangential wavelength to 310nm. The gradient was from 4 to 7 at 15 minutes tangential wavelength to 310nm.
TABLE 14 gradient elution procedure 1
TABLE 15 gradient elution procedure 2
TABLE 16 gradient elution procedure 3
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TABLE 17 gradient elution procedure 4
Table 18 gradient elution procedure 5
TABLE 19 gradient elution procedure 6
TABLE 20 gradient elution procedure 7
TABLE 21 Peak results
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The results are shown in the table and fig. 15-21, and compared with the gradient elution procedure 1-5, the optimized gradient elution procedure 7 of the invention has 9 characteristic peaks, the characteristic peaks are well separated, and the separation degree of the characteristic peaks reaches more than 1.5, so the gradient elution procedure 7 is tentatively determined for the next investigation.
(3) Different instruments
According to the method, samples are respectively taken under different brands of chromatographs (waters e2695, agilent 1290 II and Thermo U3000), the other methods are the same as the chromatographic conditions of the example 1, different instrument durability is examined, and the result shows that the RSD of the relative retention time of 9 characteristic peaks is within +/-10% of a specified value, and the optimized method has better instrument durability.
4. Characteristic peak assignment
Preparation of each control solution: taking a proper amount of Wang Baige glycoside H, wang Baige glycoside A and 2-acetyl Wang Baige glycoside A reference substances, precisely weighing, adding 50% methanol to prepare a mixed solution containing Wang Baige glycoside H40 mug, wang Baige glycoside 40 mug and 2-acetyl Wang Baige glycoside 40 mug per 1ml, shaking uniformly, and taking the mixed solution as a reference substance solution;
the reference solution and the sample solution (prepared by the method of example 1) were respectively sucked, and the sample solution was tested under the chromatographic conditions of example 1 to obtain a characteristic spectrum. Detection was performed by ultraviolet chromatography.
As shown in fig. 22 and 23, comparing with the reference solution map of the reference substance to determine that peak 3 in the characteristic map of lily (lily) is lilioside H; peak 4 is lilioside A; there was no 2-acetyl Wang Baige glycoside A chromatographic peak.
Example 10 methodological verification
1. Precision of
(1) Precision of instrument
About 1g of lily (lily) medicinal material is taken, precisely weighed, a sample solution is prepared according to the method of the example 1, sample injection is carried out for 6 times continuously, a chromatogram is recorded, the relative retention time and the RSD of the relative peak areas of 9 characteristic peaks are calculated, and the result shows that the relative retention time and the RSD of the relative peak areas of the 9 characteristic peaks are less than 3 percent. The method has good precision and meets the requirements of characteristic patterns.
About 0.2g of lily (lily) decoction piece standard decoction (freeze-dried powder) is taken, precisely weighed, a sample solution is prepared according to the method of example 3, sample injection is continuously carried out for 6 times, a chromatogram is recorded, the relative retention time and RSD (relative peak area) of 9 characteristic peaks of the invention are calculated, and the result shows that the relative retention time and RSD% of the relative peak areas of the 9 characteristic peaks are less than 2%. The method has good precision and meets the requirements of characteristic patterns,
(2) Repeatability experiments
About 1g of lily (lily) medicinal material in the same batch is taken, precisely weighed, 6 parts of test sample solution are prepared in parallel according to the method of the example 1, sample injection analysis is carried out, a chromatogram is recorded, the relative retention time and RSD (reactive) of the relative peak areas of 9 characteristic peaks are calculated, and the result shows that the relative retention time and RSD% of the relative peak areas of the 9 characteristic peaks are less than 2%. The method has good precision and meets the requirements of characteristic patterns.
About 0.2g of standard decoction (freeze-dried powder) of lily (lily) decoction pieces in the same batch is taken, precisely weighed, 6 parts of test sample solutions are prepared in parallel according to the method of example 3, sample injection analysis is carried out, a chromatogram is recorded, and the relative retention time and the RSD% of the relative peak areas of 9 characteristic peaks of the invention are calculated. The results showed that the relative retention time and relative peak area RSD% for the 9 characteristic peaks were less than 2%. The method has good precision and meets the requirements of characteristic patterns.
(3) Different personnel (middle precision)
Taking the same batch of lily (lily) medicinal materials, and performing independent operation by an experiment personnel A, an experiment personnel B and an experiment personnel C, preparing a sample solution according to the method of the embodiment 1, performing sample injection analysis under the chromatographic condition of the embodiment 1, recording a chromatogram, and calculating the relative retention time of 9 characteristic peaks and the RSD% of the relative peak areas, wherein the result shows that: the relative retention time and relative peak area RSD% for the 9 characteristic peaks was less than 3%. The comprehensive judgment is carried out, the intermediate precision (different personnel) of the method is good, and the requirements of the characteristic spectrum are met.
Taking the same batch of lily (lily) decoction piece standard decoction (freeze-dried powder), and performing independent operation by an experiment personnel A, an experiment personnel B and an experiment personnel C, preparing a sample solution according to the method of the embodiment 3, performing sample injection analysis under the chromatographic condition of the embodiment 3, recording a chromatogram, and calculating the relative retention time of 9 characteristic peaks and the RSD of the relative peak areas, wherein the result shows that: the relative retention time and relative peak area RSD% for the 9 characteristic peaks is less than 2%. The comprehensive judgment is carried out, the intermediate precision (different personnel) of the method is good, and the requirements of the characteristic spectrum are met.
2. Stability of
Taking the same lily (lily) medicinal material sample solution prepared according to the method of example 1, measuring at 0h, 4h, 8h, 12h, 16h and 24h according to the chromatographic conditions of example 1, recording a chromatogram, and calculating the relative retention time of each characteristic peak and the RSD% of the relative peak area so as to investigate the stability of the sample solution. The results show that: the relative retention time and relative peak area RSD% for the 9 characteristic peaks is less than 2%. The comprehensive judgment is carried out, the stability of the method is better, and the method meets the requirements of characteristic patterns.
Taking the same standard decoction (lyophilized powder) sample solution of lily (lily) decoction pieces prepared according to the method of example 3, respectively measuring at 0h, 4h, 8h, 12h, 16h and 24h according to the chromatographic conditions of example 3, recording a chromatogram, and calculating the relative retention time of each characteristic peak and the RSD% of the relative peak area to examine the stability of the sample solution. The results show that: the relative retention time and relative peak area RSD% for the 9 characteristic peaks is less than 2%. The comprehensive judgment is carried out, the stability of the method is better, and the method meets the requirements of characteristic patterns.
3. Specialization of
About 1g of lily (lily) medicinal material is taken, precisely weighed, a test sample solution is prepared according to the method of the embodiment 1, the test sample is taken, 50% methanol is adopted as an extraction solvent, 10 mu L of each of the test sample solution and a negative control solution (50% methanol) is precisely sucked, and the test sample solution and the negative control solution are respectively injected into a high performance liquid chromatograph, and tested according to the method of the embodiment 1, and the result shows that the negative result has no interference.
About 0.2g of lily decoction piece standard decoction (freeze-dried powder) is taken, precisely weighed, a test sample solution is prepared according to the method of the embodiment 3, the test sample adopts 50% methanol as an extraction solvent, 10 mu L of each of the test sample solution and a negative control solution (50% methanol) is precisely sucked, and the test sample solution and the negative control solution are respectively injected into a high performance liquid chromatograph, and tested according to the method of the embodiment 3, and the result shows that the test sample solution is negative without interference.
4. Durability of
(1) Different flow rates
Taking the same part of lily (lily) medicinal material sample solution prepared according to the method of example 1, respectively testing at different flow rates of 0.75mL/min, 0.80mL/min and 0.85mL/min, recording a chromatogram under the same conditions as those of example 1, and calculating the relative retention time of each characteristic peak and the RSD% of the relative peak area. The result shows that the relative retention time RSD% of 9 characteristic peaks is 0% -3.7%, and the relative peak area RSD% is 0% -8.6%, so that the influence of flow velocity fluctuation on the characteristic peaks is small.
Test was carried out at different flow rates of 0.75mL/min, 0.80mL/min and 0.85mL/min, respectively, using the same sample solution of standard decoction (lyophilized powder) of lily (lily) pieces prepared according to the method of example 3, and the other conditions were the same as those of example 1, and the chromatograms were recorded to calculate the relative retention time and RSD of the relative peak area of each characteristic peak, with the following results. The result shows that the relative retention time RSD% of 9 characteristic peaks is 0% -3.6%, and the relative peak area RSD% of 9 characteristic peaks is 0% -5.4%, so that the influence of different flow rates on the characteristic peaks is smaller.
(2) Different column temperatures
Test was carried out at 22 deg.C, 25 deg.C and 28 deg.C respectively on the same lily (lily) medicinal material sample solution prepared by the method of example 1, the other conditions were the same as those of example 1, the chromatogram was recorded, and the relative retention time of each characteristic peak and the RSD% of the relative peak area were calculated. The result shows that the relative retention time RSD% of 9 characteristic peaks is 0% -7.9%, and the relative peak area RSD% of 9 characteristic peaks is 0% -9.7%, so that the influence on the characteristic peaks under different column temperature conditions is smaller.
Test was carried out at 22 deg.c, 25 deg.c and 28 deg.c with the same standard decoction (lyophilized powder) of lily (lily) pieces prepared in the same manner as in example 3, and the other conditions were the same as in example 1, and the chromatograms were recorded to calculate the relative retention time of each characteristic peak and RSD% of the relative peak area, and the results were as follows. The result shows that the relative retention time RSD% of 9 characteristic peaks is 0% -7.8%, and the relative peak area RSD% of 9 characteristic peaks is 0% -5.7%, so that the influence on the characteristic peaks under different column temperature conditions is smaller.
(3) Different instruments
The same tester takes the lily (lily) medicinal material sample solution prepared according to the method of example 1 at different times, tests are carried out on Agilent and Waters different instruments respectively, other conditions are the same as those of example 1, a chromatogram is recorded, and the relative retention time of each characteristic peak and the RSD% of the relative peak area are calculated. The results showed that the RSD of the relative retention time of 9 characteristic peaks was within the range of 0% -8.3% of the prescribed value.
It is apparent that the above examples are given by way of illustration only and are not limiting of the embodiments. Other variations or modifications of the above teachings will be apparent to those of ordinary skill in the art. It is not necessary here nor is it exhaustive of all embodiments. While still being apparent from variations or modifications that may be made by those skilled in the art are within the scope of the invention.
Claims (10)
1. A construction method of characteristic patterns of lily medicinal materials with different origins and preparations thereof is characterized by comprising the following steps,
(1) Preparing a lily test sample solution;
(2) Detecting lily sample solution by high performance liquid chromatography, taking octadecylsilane chemically bonded silica as filler, and performing gradient elution with mobile phase comprising acetonitrile and 0.05-0.2vt% phosphoric acid water solution, wherein the gradient elution procedure comprises: 0- > 10 min- > 15 min- > 23 min- > 30 min- > 35 min, the volume percentage of acetonitrile in the mobile phase is 6% → 12% → 18% → 18% → 40% → 70%.
2. The method of claim 1, wherein step (1) comprises:
1) Extracting Bulbus Lilii sample with water to obtain extractive solution;
2) Separating solid from liquid, drying, and extracting with alcohol water solution to obtain extractive solution;
3) Separating solid from liquid, and collecting the liquid as sample solution; alternatively, step (1) includes:
1) Extracting Bulbus Lilii sample with alcohol water solution to obtain extractive solution;
2) And (3) carrying out solid-liquid separation on the extracting solution, and taking the liquid as a sample solution.
3. The method of claim 2, wherein step (1) further satisfies any one or more of the following a-F:
A. the water extraction is carried out by adding water, heating and refluxing for at least 20min, preferably 20-60min;
B. the ratio of the mass of the lily test sample to the volume of water is 0.5-2:50, and the ratio relationship of the mass to the solvent is g/mL;
C. the extraction of the alcohol aqueous solution is reflux extraction or ultrasonic extraction after adding the alcohol aqueous solution, and the extraction time is 10min-5h;
D. the alcohol aqueous solution is selected from ethanol aqueous solution or methanol aqueous solution with the volume percentage of 30-70%;
E. the solid-liquid separation is centrifugation or filtration;
F. the ratio of the mass of the lily test sample to the volume of the alcohol-water solution is 0.2-2:20, and the ratio relationship of the mass to the solvent is g/mL.
4. The method according to any one of claims 1 to 3, wherein in the step (2), the detection wavelength is 203 to 208nm before 14 to 20min, the detection wavelength is 308 to 313nm after 14 to 20min, the flow rate is 0.6 to 1.0ml/min, the column temperature is 22 to 28 ℃, and the sample injection amount is 1 to 20 μl; and/or, use a Ultimate XB-C18 column of size 4.6x250mm,5 μm.
5. The method according to any one of claims 1 to 4, further comprising the step of preparing a reference solution using at least one of Wang Baige glycoside a, wang Baige glycoside H, and 2-acetyl Wang Baige glycoside a reference, and the step of detecting the reference solution by high performance liquid chromatography according to any one of claims 1 to 5 to obtain a reference map.
6. The construction method according to claim 5, wherein the reference solution is 30-70vt% methanol aqueous solution; optionally, the concentration of the control solution is 10-100 mug/mL.
7. The method according to any one of claims 1 to 6, wherein the lily medicinal materials and preparations thereof of different base are lily medicinal materials and preparations thereof of a Paeonia ostii base, lily medicinal materials and preparations thereof of a lily base or lily medicinal materials and preparations thereof of a fine leaf lily base.
8. The construction method according to claim 7, wherein the characteristic pattern of the lily medicinal material of lily base and its preparation has 9 characteristic peaks, the peak corresponding to the Wang Baige glycoside a reference substance peak is S peak, the relative retention time of each characteristic peak and S peak is within ±10% of the prescribed value, the prescribed value is: 0.50 (peak 1), 0.62 (peak 2), 0.91 (peak 3), 1.02 (peak 5), 1.04 (peak 6), 1.30 (peak 7), 1.43 (peak 8), 1.46 (peak 9);
The lily medicinal material of the original rolling dansyl and the preparation thereof have 10 characteristic peaks, the peak corresponding to the reference peak of the Wang Baige glycoside A reference substance is an S peak, the relative retention time of each characteristic peak and the S peak is within the range of +/-10% of a specified value, and the specified value is: 0.50 (peak 1), 0.62 (peak 2), 0.91 (peak 3), 1.02 (peak 5), 1.04 (peak 6), 1.30 (peak 7), 1.43 (peak 8), 1.46 (peak 9); a chromatographic peak (peak 10) appears at the corresponding position of the chromatographic peak of the 2-acetyl Wang Baige glycoside A reference substance;
the lily medicinal material of the fine leaf lily base stock and the preparation thereof have 11 characteristic peaks, the peak corresponding to the Wang Baige glycoside A reference substance peak is an S peak, the relative retention time of each characteristic peak and the S peak is within the range of +/-10% of a specified value, and the specified value is: 0.50 (Peak 1), 0.62 (Peak 2), 0.91 (Peak 3), 1.02 (Peak 5), 1.04 (Peak 6), 1.30 (Peak 7), 1.43 (Peak 8), 1.46 (Peak 9), 1.49 (Peak 11), 1.50 (Peak 12).
9. The method for identifying lily medicinal materials with different origins and preparations thereof is characterized by comprising the step of obtaining and constructing the lily product to be identified according to the construction method of any one of claims 1-8 to obtain the characteristic spectrum of the lily product.
10. The method for identifying lily medicinal materials and preparations thereof according to claim 9, wherein the lily medicinal material is a lilium base if the lily product to be identified has a chromatographic peak at the corresponding position of the chromatographic peak of the 2-acetyl Wang Baige glycoside A reference substance, the lilium medicinal material is a lilium base if the lily product to be identified has no chromatographic peak at the corresponding position of the chromatographic peak of the 2-acetyl Wang Baige glycoside A reference substance and the relative peak area between the peak 9 and the peak 4 is less than 0.23, the lilium base is a fine-leaf lilium base if the colorless spectral peak and the relative peak area between the peak 9 and the peak 4 is greater than or equal to 0.23.
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