CN115326985B - Centipede formula granule characteristic spectrum and construction method and application thereof - Google Patents
Centipede formula granule characteristic spectrum and construction method and application thereof Download PDFInfo
- Publication number
- CN115326985B CN115326985B CN202211047665.9A CN202211047665A CN115326985B CN 115326985 B CN115326985 B CN 115326985B CN 202211047665 A CN202211047665 A CN 202211047665A CN 115326985 B CN115326985 B CN 115326985B
- Authority
- CN
- China
- Prior art keywords
- peak
- centipede
- characteristic
- mobile phase
- peaks
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 241000258920 Chilopoda Species 0.000 title claims abstract description 77
- 238000001228 spectrum Methods 0.000 title claims abstract description 46
- 238000010276 construction Methods 0.000 title claims abstract description 13
- 239000008187 granular material Substances 0.000 title description 3
- 239000002245 particle Substances 0.000 claims abstract description 50
- 238000001514 detection method Methods 0.000 claims abstract description 35
- 230000014759 maintenance of location Effects 0.000 claims abstract description 24
- 239000000463 material Substances 0.000 claims abstract description 23
- 238000004704 ultra performance liquid chromatography Methods 0.000 claims abstract description 21
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 21
- 239000012488 sample solution Substances 0.000 claims abstract description 10
- 238000009210 therapy by ultrasound Methods 0.000 claims abstract description 9
- 241000131808 Scolopendra Species 0.000 claims abstract description 8
- 238000000926 separation method Methods 0.000 claims abstract description 8
- 238000001914 filtration Methods 0.000 claims abstract description 6
- 239000012088 reference solution Substances 0.000 claims abstract description 5
- 239000000243 solution Substances 0.000 claims abstract description 5
- 239000013558 reference substance Substances 0.000 claims abstract description 4
- 238000002156 mixing Methods 0.000 claims abstract description 3
- 239000012925 reference material Substances 0.000 claims abstract description 3
- 238000012216 screening Methods 0.000 claims abstract description 3
- 238000000034 method Methods 0.000 claims description 25
- 239000000523 sample Substances 0.000 claims description 14
- 239000003814 drug Substances 0.000 claims description 13
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 12
- 238000010828 elution Methods 0.000 claims description 10
- 239000007788 liquid Substances 0.000 claims description 7
- 238000003908 quality control method Methods 0.000 claims description 6
- 229940079593 drug Drugs 0.000 claims description 5
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 4
- 238000002604 ultrasonography Methods 0.000 claims description 4
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 claims description 3
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 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
- 238000012545 processing Methods 0.000 claims description 2
- 239000000741 silica gel Substances 0.000 claims description 2
- 229910002027 silica gel Inorganic materials 0.000 claims description 2
- 239000012071 phase Substances 0.000 description 18
- 230000000052 comparative effect Effects 0.000 description 9
- 239000000047 product Substances 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- IGDFDIZIHMFYGR-UHFFFAOYSA-N jineol Chemical compound OC1=CC=CC2=CC(O)=CN=C21 IGDFDIZIHMFYGR-UHFFFAOYSA-N 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 239000000706 filtrate Substances 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 239000003643 water by type Substances 0.000 description 3
- 206010010904 Convulsion Diseases 0.000 description 2
- 241001334235 Scolopendra mutilans Species 0.000 description 2
- 208000005392 Spasm Diseases 0.000 description 2
- 230000036461 convulsion Effects 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000011835 investigation Methods 0.000 description 2
- 239000008213 purified water Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 235000017166 Bambusa arundinacea Nutrition 0.000 description 1
- 235000017491 Bambusa tulda Nutrition 0.000 description 1
- 241001330002 Bambuseae Species 0.000 description 1
- 208000006877 Insect Bites and Stings Diseases 0.000 description 1
- 208000019695 Migraine disease Diseases 0.000 description 1
- 208000002193 Pain Diseases 0.000 description 1
- 235000015334 Phyllostachys viridis Nutrition 0.000 description 1
- 241000270295 Serpentes Species 0.000 description 1
- 208000004078 Snake Bites Diseases 0.000 description 1
- 206010043376 Tetanus Diseases 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 239000011425 bamboo Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000001914 calming effect Effects 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 201000007227 lymph node tuberculosis Diseases 0.000 description 1
- 206010027599 migraine Diseases 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000003252 repetitive effect Effects 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 239000000377 silicon dioxide Substances 0.000 description 1
- 238000013112 stability test Methods 0.000 description 1
- 125000004079 stearyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000012085 test solution Substances 0.000 description 1
- 238000004809 thin layer chromatography Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—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
- G01N30/34—Control of physical parameters of the fluid carrier of fluid composition, e.g. gradient
-
- 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
-
- 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
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
-
- 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
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid 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
- G01N2030/062—Preparation extracting sample from raw material
-
- 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
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
The invention provides a centipede formula particle feature map and a construction method and application thereof, wherein the construction method comprises the following steps: (1) Decocting Scolopendra with water, and performing ultrasonic treatment and filtration to obtain reference solution of reference material; (2) Mixing centipede formula particles with water and performing ultrasonic treatment to obtain a sample solution; (3) And (3) respectively carrying out UPLC detection on the reference substance solution of the reference medicinal material and the sample solution, screening out peaks with consistent retention time, good peak shape and high separation degree as characteristic peaks according to detection results, selecting the S peak with determined components and good peak shape from the characteristic peaks, and calculating the relative retention time of other characteristic peaks relative to the S peak to obtain the centipede formula particle characteristic spectrum. The constructed characteristic spectrum provided by the invention has good repeatability, high precision and good stability, and can be used for effectively identifying the quality of centipede formula particles.
Description
Technical Field
The invention belongs to the field of traditional Chinese medicine identification, and particularly relates to a centipede formula particle characteristic spectrum and a construction method and application thereof, in particular to a centipede formula particle characteristic spectrum with high precision and a construction method and application thereof.
Background
The Scolopendra medicinal material is dried body of Scolopendra (Scolopendra, scolopendra subspinipes mutilans (Scolopendra subspinipes mutilans L. Koch)). The bamboo chips are inserted into the heads and the tails, straightened and dried after capturing in spring and summer. Has effects of calming endogenous wind, relieving spasm, dredging collaterals, relieving pain, eliminating toxic substances and resolving hard mass. Can be used for treating liver wind internal movement, spasm and convulsion, infantile convulsion, and apoplexy
Hemiplegia, tetanus, rheumatism, migraine, sore, scrofula, snake and insect bite and the like. According to the examination, scolopendra was originally in Shen nong Ben Cao Jing (Shen nong's herbal), and is listed as the lower grade, pungent in flavor, warm in nature and toxic. The current legal standards only make standard restrictions on the properties, examination and extract items of centipedes. In the prior art, the centipede formula particles and medicinal materials are less researched. Some researchers develop and research the establishment of the protein fingerprint of the centipede medicinal material, and measure the nucleotide components in the centipede or measure a certain component of centipede particles.
CN105004809B discloses a quality detection method of centipede medicinal materials, which uses 3, 8-dihydroxyquinoline in centipede as an index component, and adopts high performance liquid chromatography or thin layer chromatography to detect the quality of the centipede medicinal materials. According to the technical scheme, 3, 8-dihydroxyquinoline is used as an index component for centipede medicinal material quality detection for the first time. The method has the advantages of simple operation, controllable conditions, strong specificity, good stability and reproducibility, can effectively make up for the defects of the centipede medicinal material quality detection method in Chinese pharmacopoeia, and has strong practicability.
However, the researches on the substance basis of the centipede medicinal materials or the formula particles are all aimed at, and the centipede formula particles are prepared by the steps of extraction, concentration, drying, preparation and the like of the centipede medicinal materials due to the fact that the original appearance characteristics of the medicinal materials are lost, and the substance basis of the centipede medicinal materials and the substance basis of the medicinal materials are greatly different, so that a quality control method for the centipede formula particles and the whole medicinal materials is not available at the same time at present.
Because the quality control method for centipede formula particles is less researched at present. Therefore, how to provide a characteristic spectrum of the centipede formula particle and effectively control the quality of the centipede formula particle becomes a problem to be solved urgently.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide a centipede formula particle characteristic spectrum and a construction method and application thereof, in particular to a centipede formula particle characteristic spectrum with high precision and a construction method and application thereof. The constructed characteristic spectrum provided by the invention has good repeatability, high precision and good stability, and can be used for effectively identifying the quality of centipede formula particles.
In order to achieve the aim of the invention, the invention adopts the following technical scheme:
in a first aspect, the invention provides a method for constructing a centipede formula particle characteristic spectrum, which comprises the following steps:
(1) Decocting Scolopendra with water, and performing ultrasonic treatment and filtration to obtain reference solution of reference material;
(2) Mixing centipede formula particles with water and performing ultrasonic treatment to obtain a sample solution;
(3) And (3) respectively carrying out UPLC detection on the reference substance solution of the reference medicinal material and the sample solution, screening out peaks with consistent retention time, good peak shape and high separation degree as characteristic peaks according to detection results, selecting the S peak with determined components and good peak shape from the characteristic peaks, and calculating the relative retention time of other characteristic peaks relative to the S peak to obtain the centipede formula particle characteristic spectrum.
The step (1) and the step (2) do not distinguish the sequence.
According to the method, the centipede formula particles are extracted and are compared with centipede medicinal materials, and the characteristic spectrogram is constructed together, so that the obtained characteristic spectrogram can be used for effectively identifying the centipede formula particles; and the characteristic spectrum obtained by adopting a specific method to carry out UPLC detection has good repeatability, high precision and good durability.
Preferably, the feed liquid ratio of the centipede medicinal material to water in the step (1) is 1 (15-25) g/mL.
Preferably, the time of the ultrasonic treatment in the step (1) is 25-35min.
Preferably, the feed liquid ratio of the centipede formula particles to water in the step (2) is 1 (80-120) g/mL.
Preferably, the time of the ultrasonic treatment in the step (2) is 25-35min.
The ratio of the centipede medicinal material to water may be 1:15g/mL, 1:16g/mL, 1:17g/mL, 1:18g/mL, 1:19g/mL, 1:20g/mL, 1:21g/mL, 1:22g/mL, 1:23g/mL, 1:24g/mL, or 1:25g/mL, etc., the time of the ultrasound may be 25min, 26min, 27min, 28min, 29min, 30min, 31min, 32min, 33min, 34min, 35min, etc., the ratio of the centipede formula particles to water may be 1:80g/mL, 1:90g/mL, 1:100g/mL, 1:110g/mL, 1:120g/mL, etc., the time of the ultrasound may be 25min, 26min, 27min, 28min, 29min, 30min, 31min, 32min, 33min, 34min, 35min, etc., but the above numerical values are not limited thereto.
Preferably, the packing material of the chromatographic column detected by UPLC in the step (3) is octadecylsilane chemically bonded silica.
Preferably, the mobile phase detected by the UPLC in the step (3) includes a mobile phase a and a mobile phase B, wherein the mobile phase a is acetonitrile, and the mobile phase B is water.
Preferably, the flow rate of the UPLC detection in the step (3) is 0.28-0.32mL/min, the column temperature is 32-38 ℃, wherein the flow rate can be 0.28mL/min, 0.29mL/min, 0.3mL/min, 0.31mL/min or 0.32mL/min, and the like, the column temperature can be 32 ℃, 33 ℃, 34 ℃, 35 ℃, 36 ℃, 37 ℃ or 38 ℃, and the like, but the UPLC detection method is not limited to the above-listed values, and other non-listed values in the above-listed value range are equally applicable.
Preferably, the elution procedure for the UPLC detection in step (3) is as follows:
at 0-3min, the volume fraction of the mobile phase A is 0, and the volume fraction of the mobile phase B is 100%;
at 3-8min, the volume fraction of the mobile phase A is changed from 0 to 2% at uniform speed, and the volume fraction of the mobile phase B is changed from 100% to 98% at uniform speed;
at 8-15min, the volume fraction of the mobile phase A is changed from 2% to 23% at uniform speed, and the volume fraction of the mobile phase B is changed from 98% to 77% at uniform speed;
at 15-18min, the volume fraction of mobile phase A was 23% and the volume fraction of mobile phase B was 77%.
Preferably, the wavelength of the UPLC detection is 254nm.
The specific elution program can effectively distinguish components in the sample, has high peak shape separation degree and obvious peak, can obtain characteristic peaks with good peak shapes, improves the accuracy of detection and characteristic patterns, can obviously shorten the elution time, and improves the detection efficiency.
In a second aspect, the invention provides a centipede formula particle characteristic spectrum obtained by the construction method, which is characterized in that the number of characteristic peaks in the characteristic spectrum is 7, the characteristic peaks are ordered from small to large according to retention time, the 6 # peak is an S peak, and the relative retention time of the 1 # peak, the 2 # peak, the 3 # peak, the 4 # peak, the 5 # peak and the 7 # peak is 0.24+/-10%, 0.27+/-10%, 0.32+/-10%, 0.70+/-10%, 0.76+/-10% and 1.92+/-10% respectively.
The peak component No. 6 is tryptophan.
In a third aspect, the invention provides an application of the centipede formula particle characteristic spectrum in centipede medicine quality control.
In a fourth aspect, the invention also provides a quality control method of centipede medicines, wherein different batches of centipede medicines are detected by adopting the UPLC detection method, the detection result is subjected to median processing, and a standard characteristic spectrum is obtained by combining the characteristic spectrum; detecting a sample to be detected by adopting the UPLC detection method, judging a characteristic peak according to the characteristic spectrum, and then judging the similarity, and judging whether the sample to be detected is a qualified product or a unqualified product;
in the similarity judgment, calculating the similarity between the characteristic peak of the sample to be detected and the characteristic peak in the standard characteristic map; and if the similarity is not lower than 0.85, the sample to be detected is a qualified product, and if the similarity is lower than 0.85, the sample to be detected is a disqualified product.
The centipede medicine is centipede formula particles.
Compared with the prior art, the invention has the following beneficial effects:
the invention provides a centipede formula particle characteristic spectrum, which is obtained by extracting centipede formula particles, comparing the centipede formula particles with centipede medicinal materials, and constructing a characteristic spectrum together, wherein the obtained characteristic spectrum can be used for effectively identifying the centipede formula particles; the characteristic spectrum obtained by adopting a specific method to carry out UPLC detection has good repeatability, high precision and good durability; the components in the sample can be effectively distinguished through a specific elution program, the peak shape separation degree is high, the peak is obvious, the characteristic peak with good peak shape can be obtained, the accuracy of detection and characteristic spectrum is improved, the elution time can be obviously shortened, and the detection efficiency is improved.
Drawings
FIG. 1 is a characteristic map of example 1;
FIG. 2 is a characteristic spectrum of comparative example 1;
FIG. 3 is a characteristic spectrum of comparative example 2;
FIG. 4 is a characteristic spectrum of comparative example 3;
FIG. 5 is a graph of liquid phase results of a test for testing centipede formula particles.
Detailed Description
The technical scheme of the invention is further described by the following specific embodiments. It will be apparent to those skilled in the art that the examples are merely to aid in understanding the invention and are not to be construed as a specific limitation thereof.
The instruments and reagents used in the examples below were as follows:
instrument: waters ACQUITY
An H-Class ultra-high performance liquid chromatograph, a TUV Detector ultraviolet Detector, an Empower 3 chromatographic workstation; ML204T electronic balance (mertrehler tolido); JA1002 electronic balance (Shanghai Pu Chun metering instruments limited); MSA6.6S-OCE-DM electronic balance (Sidoriko instruments, inc.); KQ-100DE numerical control ultrasonic waveCleaning machine (Kunshan ultrasonic instruments Co., ltd.).
Chromatographic column: waters
T3(2.1×100mm，1.6μm)；
Reagent: acetonitrile (merck, inc.); the water is distilled water (Chen's); the other reagents were all analytically pure.
Example 1
The embodiment provides a centipede formula particle characteristic spectrum, which is constructed by the following steps:
preparation of reference solution: taking 2.0g of centipede reference medicine, placing the centipede reference medicine into a beaker, adding 40mL of purified water, decocting for 50 minutes, filtering, evaporating filtrate, adding 20mL of water, dissolving in a conical flask, sealing, performing ultrasonic treatment (power 500W, frequency 40 kHz) for 30 minutes, cooling, filtering, and taking the subsequent filtrate as a reference substance solution of the reference medicine.
Preparation of test solution: taking appropriate amount of Scolopendra formulation granule, grinding, taking 0.2g, precisely weighing, placing into conical flask, precisely adding purified water 20mL, ultrasonic treating (power 500W, frequency 40 kHz) for 30min, shaking, filtering, and collecting subsequent filtrate.
Assay: precisely sucking 1 μl of each of the reference solution and the sample solution, and measuring with ultra-high liquid chromatograph.
Wherein the chromatographic conditions are: octadecyl bonded silica gel is used as filler (column length is 100mm, inner diameter is 2.1mm, and particle diameter is 1.6 μm); acetonitrile is taken as a mobile phase A, water is taken as a mobile phase B, and gradient elution is carried out according to the specifications in the following table; the flow rate was 0.3mL per minute; the column temperature is 35 ℃; the detection wavelength is 254nm;
a characteristic map is obtained as shown in fig. 1. The sample chromatograph shows 7 characteristic peaks and corresponds to the retention time of 7 characteristic peaks in the reference chromatographic peaks of the control medicinal material, wherein the peak 6 is identified as tryptophan; the relative retention time of characteristic peaks 1 to 5, 7 was calculated with peak number 6 as S peak, and the relative retention time was within.+ -. 10% of the prescribed value. The specified value is: 0.24 (Peak 1), 0.27 (Peak 2), 0.32 (Peak 3), 0.70 (Peak 4), 0.76 (Peak 5), and 1.92 (Peak 7) to obtain the characteristic map.
Comparative example 1
This comparative example provides a centipede formula particle profile, which is identical to example 1 except that the elution parameters are as follows:
flow rate: 0.4mL/min, gradient elution procedure was as follows:
the results are shown in FIG. 2.
It was found that under the above conditions, the chromatographic peak was earlier in time and less in peak, and it was difficult to distinguish the feature fractions.
Comparative example 2
This comparative example provides a centipede formula particle profile, consistent with example 1 except that the gradient elution procedure is as follows:
the results are shown in FIG. 3.
It was found that the chromatographic peak separation was poor within 0.5-2min, and it was difficult to distinguish the characteristic peaks.
Comparative example 3
This comparative example provides a centipede formula particle profile, which is identical to example 1 except that the elution parameters are as follows:
methanol is taken as a mobile phase A, water is taken as a mobile phase B, and the gradient elution procedure is as follows:
the results are shown in FIG. 4.
It can be found that the front chromatographic peak is relatively concentrated in the figure and the separation effect is poor.
According to the results, the invention can effectively distinguish the characteristic components in the sample by adopting the specific mobile phase and the elution program, so that the peak shape separation degree is high, the peak is obvious, the characteristic peak with better peak shape can be obtained, and the accuracy of detection and characteristic spectrum is improved; meanwhile, the peak time is short, and the detection efficiency is high.
Repeatability test:
the procedure of example 1 was repeated six times and the relative retention time was calculated as follows:
it can be found that the relative retention time RSD of each characteristic peak is between 0 and 0.7%, which indicates that the method provided by the invention has good repeatability.
Intermediate precision test:
6 parts of centipede formula particles were taken and measured as provided in example 1 (the ultra performance liquid chromatograph was replaced by another Waters UPLC H-Class, TUV detector) and the relative retention time of each characteristic peak was calculated as follows:
the relative retention time RSD of each characteristic peak can be found to be in the range of 0-0.2%, which indicates that the intermediate precision of the characteristic spectrum is better; comparison with the repetitive sample shows that the precision of the characteristic map is better.
Stability test:
the sample solutions prepared in example 1 were detected at the completion of preparation for the 0 th, 2 nd, 4 th, 6 th, 8 th, 10 th, 12 th and 24 th hours, characteristic peaks in the sample solutions were analyzed, and the relative peak areas and relative retention times were calculated, with the following results:
it can be found that the chemical components in the test sample solution can be kept stable within 24 hours, and the relative retention time RSD of each characteristic peak is within 0-1.8%, which shows that the method provided by the invention is stable and reliable.
Column temperature investigation:
based on the method of example 1, the characteristic patterns at a column temperature of 32℃and 35℃and 38℃were examined, respectively, and the results were as follows:
from the above data, it can be seen that the relative retention time of 7 peaks of the characteristic spectrum obtained at 35 ℃ is within + -10% of the relative retention time of 7 peaks of the characteristic spectrum obtained at other two temperatures, which indicates that the method has better tolerance to column temperature.
Flow rate investigation:
based on the method of example 1, the characteristic patterns at different flow rates (0.28 mL/min, 0.30mL/min and 0.32 mL/min) were examined, and the results were as follows:
from the above data, it can be seen that the relative retention time of 7 peaks of the characteristic spectrum obtained by the other two flow rates is within + -10% of the relative retention time of the characteristic spectrum obtained by the other two flow rates by taking the relative retention time of the characteristic spectrum obtained by 0.30mL/min as a reference, which shows that the method has better tolerance to the flow rates.
Centipede formula particle detection experiment:
five batches of centipede formula particles are taken, detected respectively according to the method in the embodiment 1, the detection patterns are fitted to obtain standard characteristic patterns, characteristic peaks are judged according to the characteristic patterns obtained in the embodiment 1, the similarity is calculated, and the results are shown in fig. 5 and the following table:
the similarity between the characteristic spectrum of the five batches of centipede formula particles and the contrast characteristic spectrum is 0.986-0.999, which is higher than 0.85, and the five batches of centipede formula particles are all qualified products.
The detection test fully proves that the characteristic spectrum and the construction method thereof provided by the invention can effectively judge the quality of centipede formula particles, and a novel method is provided for judging the quality of centipede medicines.
The applicant states that the invention is described by the above examples to illustrate the characteristic spectrum of the centipede formula particles of the invention and the construction method and application thereof, but the invention is not limited to the above examples, i.e. the invention is not necessarily limited to practice depending on the above examples. It should be apparent to those skilled in the art that any modification of the present invention, equivalent substitution of raw materials for the product of the present invention, addition of auxiliary components, selection of specific modes, etc., falls within the scope of the present invention and the scope of disclosure.
The preferred embodiments of the present invention have been described in detail above, but the present invention is not limited to the specific details of the above embodiments, and various simple modifications can be made to the technical solution of the present invention within the scope of the technical concept of the present invention, and all the simple modifications belong to the protection scope of the present invention.
In addition, the specific features described in the above embodiments may be combined in any suitable manner, and in order to avoid unnecessary repetition, various possible combinations are not described further.
Claims (9)
1. The construction method of the centipede formula particle characteristic spectrum is characterized by comprising the following steps of:
(1) Decocting Scolopendra with water, and performing ultrasonic treatment and filtration to obtain reference solution of reference material;
(2) Mixing centipede formula particles with water and performing ultrasonic treatment to obtain a sample solution;
(3) Respectively carrying out UPLC detection on reference substance solutions of reference medicinal materials and sample solutions, screening out peaks with consistent retention time, good peak shape and high separation degree as characteristic peaks according to detection results, selecting the characteristic peaks as S peaks with determined components and good peak shape, and calculating the relative retention time of other characteristic peaks relative to the S peaks to obtain a centipede formula particle characteristic spectrum;
step (1) and step (2) do not distinguish the sequence;
the filler of the chromatographic column detected by UPLC in the step (3) is octadecylsilane chemically bonded silica gel;
the mobile phase detected by UPLC in the step (3) comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is acetonitrile, and the mobile phase B is water;
the elution procedure for the UPLC detection in step (3) is as follows:
at 0-3min, the volume fraction of the mobile phase A is 0, and the volume fraction of the mobile phase B is 100%;
at 3-8min, the volume fraction of the mobile phase A is changed from 0 to 2% at uniform speed, and the volume fraction of the mobile phase B is changed from 100% to 98% at uniform speed;
at 8-15min, the volume fraction of the mobile phase A is changed from 2% to 23% at uniform speed, and the volume fraction of the mobile phase B is changed from 98% to 77% at uniform speed;
at 15-18min, the volume fraction of mobile phase A is 23% and the volume fraction of mobile phase B is 77%;
in the characteristic spectrum, the number of characteristic peaks is 7, the characteristic peaks are sequenced from small to large according to retention time, the number 6 peak is an S peak, and the relative retention time of the number 1 peak, the number 2 peak, the number 3 peak, the number 4 peak, the number 5 peak and the number 7 peak is 0.24+/-10%, 0.27+/-10%, 0.32+/-10%, 0.70+/-10%, 0.76+/-10% and 1.92+/-10% respectively;
the peak component No. 6 is tryptophan.
2. The construction method according to claim 1, wherein the feed liquid ratio of the centipede medicinal material to water in the step (1) is 1 (15-25) g/mL.
3. The method of claim 1, wherein the time of the ultrasound in step (1) is 25-35min.
4. The method of claim 1, wherein the ratio of feed liquid of the centipede formula particles to water in step (2) is 1 (80-120) g/mL.
5. The method of claim 1, wherein the time of the ultrasound in step (2) is 25-35min.
6. The method according to claim 1, wherein the UPLC detection in the step (3) has a flow rate of 0.28-0.32mL/min and a column temperature of 32-38 ℃.
7. The method of claim 1, wherein the UPLC detection wavelength is 254nm.
8. Use of a method for constructing a characteristic spectrum of centipede formula particles according to any one of claims 1-7 in centipede medicine quality control.
9. A quality control method of centipede medicines, which is characterized in that the UPLC detection method of any one of claims 1-7 is adopted for detection of different batches of centipede medicines, the detection result is subjected to median processing, and a standard characteristic spectrum is obtained by combining with the characteristic spectrum; detecting a sample to be detected by adopting the UPLC detection method of any one of claims 1-7, judging a characteristic peak according to the characteristic spectrum, and then judging the similarity, wherein the sample to be detected is judged to be qualified or unqualified;
in the similarity judgment, calculating the similarity between the characteristic peak of the sample to be detected and the characteristic peak in the standard characteristic map; the sample to be detected is qualified when the similarity is not lower than 0.85, and is unqualified when the similarity is lower than 0.85;
the centipede medicine is centipede formula particles.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211047665.9A CN115326985B (en) | 2022-08-30 | 2022-08-30 | Centipede formula granule characteristic spectrum and construction method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211047665.9A CN115326985B (en) | 2022-08-30 | 2022-08-30 | Centipede formula granule characteristic spectrum and construction method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115326985A CN115326985A (en) | 2022-11-11 |
| CN115326985B true CN115326985B (en) | 2023-06-02 |
Family
ID=83927237
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211047665.9A Active CN115326985B (en) | 2022-08-30 | 2022-08-30 | Centipede formula granule characteristic spectrum and construction method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115326985B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116124921A (en) * | 2022-12-06 | 2023-05-16 | 劲牌持正堂药业有限公司 | Construction method and quality control method of characteristic spectrogram of centipede formula particles |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105004809A (en) * | 2015-07-13 | 2015-10-28 | 鲁南制药集团股份有限公司 | Centipede medicinal material quality control method |
| CN106404942A (en) * | 2016-08-29 | 2017-02-15 | 广州品红制药有限公司 | Kangshen granule fingerprint construction method and standard fingerprint thereof |
| CN113960203A (en) * | 2021-10-22 | 2022-01-21 | 中国科学院上海药物研究所 | Method for detecting characteristic spectrum of tianlong formula particles |
| CN114894934A (en) * | 2022-05-17 | 2022-08-12 | 江阴天江药业有限公司 | Construction method and application of UPLC (unified Power LC) characteristic spectrum of scorpion medicinal material and formula granules thereof |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008112561A1 (en) * | 2007-03-09 | 2008-09-18 | Pfantastic Medical Research Institute, Llc | Combination therapy for alleviating pain-related conditions |
-
2022
- 2022-08-30 CN CN202211047665.9A patent/CN115326985B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105004809A (en) * | 2015-07-13 | 2015-10-28 | 鲁南制药集团股份有限公司 | Centipede medicinal material quality control method |
| CN106404942A (en) * | 2016-08-29 | 2017-02-15 | 广州品红制药有限公司 | Kangshen granule fingerprint construction method and standard fingerprint thereof |
| CN113960203A (en) * | 2021-10-22 | 2022-01-21 | 中国科学院上海药物研究所 | Method for detecting characteristic spectrum of tianlong formula particles |
| CN114894934A (en) * | 2022-05-17 | 2022-08-12 | 江阴天江药业有限公司 | Construction method and application of UPLC (unified Power LC) characteristic spectrum of scorpion medicinal material and formula granules thereof |
Non-Patent Citations (2)
| Title |
|---|
| Development of a novel UHPLC-UV combined with UHPLC-QTOF/MSfinge rprint method for the comprehensive evaluation of Nao-Luo-Xin-Tong:multi-wavelength setting based on traditional Chinese medicinal prescription composition;Lina Wang 等;Analytical Methods;第11卷;6092– 6102 * |
| 蜈蚣配方颗粒剂的研究;中国优秀硕士学位论文全文数据库 医药卫生科技(第04期);E057-7 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115326985A (en) | 2022-11-11 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN113702541B (en) | Poria cocos medicinal material characteristic spectrum construction method and poria cocos triterpene component detection method | |
| CN102520079B (en) | Method for rapidly measuring content of solanesol in tobaccos by using UPLC (Ultra Performance Liquid Chromatography) | |
| CN115326985B (en) | Centipede formula granule characteristic spectrum and construction method and application thereof | |
| CN108152399B (en) | Construction and detection method of UPLC (ultra performance liquid chromatography) characteristic spectrum of semen boitae medicinal material | |
| CN110133158B (en) | HPLC fingerprint detection method of wine steamed coptis chinensis | |
| CN110031564B (en) | Quality detection method of natural plant anticoccidial feed additive based on HPLC fingerprint | |
| CN109655558B (en) | Method for detecting effective part group of periploca forrestii schltr | |
| CN106290603B (en) | A kind of method and application detecting inorganic anion in plant, organic acid and three kinds of phytochemicals simultaneously using Vavle switching method | |
| CN113759037B (en) | Characteristic spectrum of formula granules of semen lepidii and/or semen lepidii as well as construction method and identification method thereof | |
| CN114942291B (en) | Quality detection method of yang-suppressing yin-nourishing particles | |
| CN115144522B (en) | Control method for comprehensively controlling quality of gecko formula particles | |
| CN113759011B (en) | Method for establishing characteristic spectrum of starwort root and preparation thereof | |
| CN115372534A (en) | Folium artemisiae argyi and construction method of characteristic map of preparation of folium artemisiae argyi, characteristic map and application | |
| CN114563497A (en) | Quantitative fingerprint detection method for bupleurum tenue capsule components | |
| CN110031533B (en) | Method for separating and detecting polyphenol substances in mulberry leaves by combining solid-phase extraction and capillary electrophoresis technology | |
| Luo et al. | Determination of active ingredients in Chinese medicine Danning Tablets using dispersion solid-phase extraction by molecular imprinting nanomaterials coupled with HPLC-DAD | |
| CN111257479A (en) | Method for extracting and analyzing trivalent chromium and hexavalent chromium in water body sediment | |
| CN115078588B (en) | Aspongopus and quality evaluation method of processed product thereof | |
| CN114965739B (en) | Method for detecting kadsura pepper stem and preparation and quality control method thereof | |
| CN115266997B (en) | Construction method and application of puffball medicinal material and formula particle characteristic spectrum thereof | |
| CN114965758B (en) | Construction method and application of characteristic spectrum of alisma orientale or pharmaceutical preparation thereof | |
| CN115308331B (en) | Method for measuring content of 5 ingredients in oldenlandia standard decoction freeze-dried powder or formula granules by adopting one-measurement-multiple-evaluation method | |
| CN115436520B (en) | Construction and detection method of luffa UPLC characteristic spectrum | |
| CN118330097B (en) | Method for identifying rheum officinale in rheum officinale cathartichoke tablets | |
| CN115389652B (en) | Method for establishing fingerprinting of rhizoma phragmitis medicinal material |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |