CN115343376A - Method for detecting or separating components of stomach-clearing coptis tablet - Google Patents
Method for detecting or separating components of stomach-clearing coptis tablet Download PDFInfo
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- CN115343376A CN115343376A CN202110529261.2A CN202110529261A CN115343376A CN 115343376 A CN115343376 A CN 115343376A CN 202110529261 A CN202110529261 A CN 202110529261A CN 115343376 A CN115343376 A CN 115343376A
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- 238000000034 method Methods 0.000 title claims abstract description 61
- 241000218202 Coptis Species 0.000 title abstract description 15
- 235000002991 Coptis groenlandica Nutrition 0.000 title abstract description 15
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- VKJGBAJNNALVAV-UHFFFAOYSA-M Berberine chloride (TN) Chemical compound [Cl-].C1=C2CC[N+]3=CC4=C(OC)C(OC)=CC=C4C=C3C2=CC2=C1OCO2 VKJGBAJNNALVAV-UHFFFAOYSA-M 0.000 claims abstract description 23
- 238000004128 high performance liquid chromatography Methods 0.000 claims abstract description 22
- DGLDSNPMIYUWGN-OOJQBDKLSA-N (13as)-3,10-dimethoxy-7-methyl-6,8,13,13a-tetrahydro-5h-isoquinolino[2,1-b]isoquinolin-7-ium-2,11-diol;chloride Chemical compound [Cl-].C1CC2=CC(OC)=C(O)C=C2[C@H]2[N+]1(C)CC(C=C(C(=C1)O)OC)=C1C2 DGLDSNPMIYUWGN-OOJQBDKLSA-N 0.000 claims abstract description 21
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- 210000002784 stomach Anatomy 0.000 claims abstract description 8
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- 238000001514 detection method Methods 0.000 claims description 23
- RLQYRXCUPVKSAW-UHFFFAOYSA-M 2,3,9,10-tetramethoxy-5,6-dihydroisoquinolino[2,1-b]isoquinolin-7-ium;chloride Chemical compound [Cl-].COC1=C(OC)C=C2CC[N+]3=CC4=C(OC)C(OC)=CC=C4C=C3C2=C1 RLQYRXCUPVKSAW-UHFFFAOYSA-M 0.000 claims description 21
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- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 6
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- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical group CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims description 3
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- IBFYXTRXDNAPMM-FZEIBHLUSA-N Geniposide Natural products COC(=O)C1=CO[C@@H](O[C@H]2O[C@@H](CO)[C@H](O)[C@@H](O)[C@@H]2O)[C@H]2[C@@H]1CC=C2CO IBFYXTRXDNAPMM-FZEIBHLUSA-N 0.000 abstract description 15
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- DTOUWTJYUCZJQD-UJERWXFOSA-N Forsythiaside Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1OC[C@@H]1[C@@H](OC(=O)\C=C\C=2C=C(O)C(O)=CC=2)[C@H](O)[C@@H](O)[C@H](OCCC=2C=C(O)C(O)=CC=2)O1 DTOUWTJYUCZJQD-UJERWXFOSA-N 0.000 abstract description 12
- 238000011084 recovery Methods 0.000 abstract description 4
- YKRGDOXKVOZESV-WRJNSLSBSA-N Paeoniflorin Chemical compound C([C@]12[C@H]3O[C@]4(O)C[C@](O3)([C@]1(C[C@@H]42)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)C)OC(=O)C1=CC=CC=C1 YKRGDOXKVOZESV-WRJNSLSBSA-N 0.000 abstract description 2
- 230000002349 favourable effect Effects 0.000 abstract description 2
- YKRGDOXKVOZESV-UHFFFAOYSA-N paeoniflorin Natural products O1C(C)(C2(CC34)OC5C(C(O)C(O)C(CO)O5)O)CC3(O)OC1C24COC(=O)C1=CC=CC=C1 YKRGDOXKVOZESV-UHFFFAOYSA-N 0.000 abstract description 2
- FUKUFMFMCZIRNT-UHFFFAOYSA-N hydron;methanol;chloride Chemical compound Cl.OC FUKUFMFMCZIRNT-UHFFFAOYSA-N 0.000 description 33
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- QIMGUQIHCNDUKU-UHFFFAOYSA-N 2-[[6-[2-(3,4-dihydroxyphenyl)ethoxy]-3,4,5-trihydroxyoxan-2-yl]methoxy]-6-methyloxane-3,4,5-triol Chemical compound OC1C(O)C(O)C(C)OC1OCC1C(O)C(O)C(O)C(OCCC=2C=C(O)C(O)=CC=2)O1 QIMGUQIHCNDUKU-UHFFFAOYSA-N 0.000 description 9
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 9
- PTPHDVKWAYIFRX-UHFFFAOYSA-N Palmatine Natural products C1C2=C(OC)C(OC)=CC=C2C=C2N1CCC1=C2C=C(OC)C(OC)=C1 PTPHDVKWAYIFRX-UHFFFAOYSA-N 0.000 description 9
- RBBVPNQTBKHOEQ-KKSFZXQISA-O Phellodendrine Chemical compound C1CC2=CC(OC)=C(O)C=C2[C@H]2[N@+]1(C)CC(C=C(C(=C1)O)OC)=C1C2 RBBVPNQTBKHOEQ-KKSFZXQISA-O 0.000 description 9
- QISXPYZVZJBNDM-UHFFFAOYSA-N berberine Natural products COc1ccc2C=C3N(Cc2c1OC)C=Cc4cc5OCOc5cc34 QISXPYZVZJBNDM-UHFFFAOYSA-N 0.000 description 9
- YBHILYKTIRIUTE-UHFFFAOYSA-N berberine Chemical compound C1=C2CC[N+]3=CC4=C(OC)C(OC)=CC=C4C=C3C2=CC2=C1OCO2 YBHILYKTIRIUTE-UHFFFAOYSA-N 0.000 description 9
- 229940093265 berberine Drugs 0.000 description 9
- 229930189432 forsythoside Natural products 0.000 description 9
- QUCQEUCGKKTEBI-UHFFFAOYSA-N palmatine Chemical compound COC1=CC=C2C=C(C3=C(C=C(C(=C3)OC)OC)CC3)[N+]3=CC2=C1OC QUCQEUCGKKTEBI-UHFFFAOYSA-N 0.000 description 9
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- 239000012498 ultrapure water Substances 0.000 description 7
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- GMBQZIIUCVWOCD-WWASVFFGSA-N Sarsapogenine Chemical compound O([C@@H]1[C@@H]([C@]2(CC[C@@H]3[C@@]4(C)CC[C@H](O)C[C@H]4CC[C@H]3[C@@H]2C1)C)[C@@H]1C)[C@]11CC[C@H](C)CO1 GMBQZIIUCVWOCD-WWASVFFGSA-N 0.000 description 2
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- 244000286838 Eclipta prostrata Species 0.000 description 1
- 244000111489 Gardenia augusta Species 0.000 description 1
- XJMPAUZQVRGFRE-SCHFUKFYSA-N Gardenoside Natural products O=C(OC)C=1[C@H]2[C@H]([C@H](O[C@H]3[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O3)OC=1)[C@@](O)(CO)C=C2 XJMPAUZQVRGFRE-SCHFUKFYSA-N 0.000 description 1
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- 206010034829 Pharyngeal oedema Diseases 0.000 description 1
- 241000972673 Phellodendron amurense Species 0.000 description 1
- RTMWIZOXNKJHRE-UHFFFAOYSA-N Tigogenin Natural products CC1COC2CC(C)(OC12)C3CCC4C5CCC6CC(O)CCC6(C)C5CCC34C RTMWIZOXNKJHRE-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 208000002399 aphthous stomatitis Diseases 0.000 description 1
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- LPLVUJXQOOQHMX-QWBHMCJMSA-N glycyrrhizinic acid Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@H](O[C@@H]1O[C@@H]1C([C@H]2[C@]([C@@H]3[C@@]([C@@]4(CC[C@@]5(C)CC[C@@](C)(C[C@H]5C4=CC3=O)C(O)=O)C)(C)CC2)(C)CC1)(C)C)C(O)=O)[C@@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O LPLVUJXQOOQHMX-QWBHMCJMSA-N 0.000 description 1
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- XJMPAUZQVRGFRE-AYDWLWLASA-N methyl (1s,4as,7s,7as)-7-hydroxy-7-(hydroxymethyl)-1-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-4a,7a-dihydro-1h-cyclopenta[c]pyran-4-carboxylate Chemical compound O([C@@H]1OC=C([C@@H]2[C@H]1[C@](C=C2)(O)CO)C(=O)OC)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O XJMPAUZQVRGFRE-AYDWLWLASA-N 0.000 description 1
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- RLLPVAHGXHCWKJ-UHFFFAOYSA-N permethrin Chemical compound CC1(C)C(C=C(Cl)Cl)C1C(=O)OCC1=CC=CC(OC=2C=CC=CC=2)=C1 RLLPVAHGXHCWKJ-UHFFFAOYSA-N 0.000 description 1
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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
- G01N30/30—Control of physical parameters of the fluid carrier of 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
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/32—Control of physical parameters of the fluid carrier of pressure or speed
-
- 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
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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
- G01N30/62—Detectors specially adapted therefor
- G01N30/74—Optical detectors
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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
- G01N30/86—Signal analysis
- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
- G01N30/8679—Target compound analysis, i.e. whereby a limited number of peaks is analysed
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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
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/30—Control of physical parameters of the fluid carrier of temperature
- G01N2030/3007—Control of physical parameters of the fluid carrier of temperature same temperature for whole column
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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
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/32—Control of physical parameters of the fluid carrier of pressure or speed
- G01N2030/324—Control of physical parameters of the fluid carrier of pressure or speed speed, flow rate
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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)
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- Immunology (AREA)
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- Spectroscopy & Molecular Physics (AREA)
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Abstract
The invention discloses a method for detecting or separating components of a Chinese medicinal composition for clearing stomach, which comprises the step of detecting a test solution containing the Chinese medicinal composition for clearing stomach by using high performance liquid chromatography; in the high performance liquid chromatography, the adopted mobile phase A is acetonitrile, and the mobile phase B is 0.09-0.11% phosphoric acid aqueous solution; the gradient elution conditions for the mobile phases a and B are shown in the present invention. The method for detecting or separating the components of the stomach-clearing coptis tablet can simultaneously measure the content of 6 effective components such as geniposide, phellodendrine hydrochloride, paeoniflorin, forsythoside A, baicalin, berberine hydrochloride and the like, and can simply, quickly, effectively, accurately and reliably separate and detect various effective components in the stomach-clearing coptis tablet, so that the quality of the stomach-clearing coptis tablet is controlled, the method has good repeatability, the recovery rate meets the requirements, the cost is lower, and the method is more favorable for industrial application.
Description
Technical Field
The invention relates to the field of quality analysis, in particular to a method for detecting or separating components of a stomach-clearing coptis tablet.
Background
The QINGWEIHUANG tablet is a Chinese medicinal preparation, has effects of clearing stomach-fire, removing toxic substance and relieving swelling, and can be used for treating aphtha of the mouth and tongue, gum and throat swelling and pain caused by excessive lung-stomach fire. The composition of QINGWEIHUANG tablet comprises Coptidis rhizoma, gypsum Fibrosum, radix Platycodi, glycyrrhrizae radix, rhizoma anemarrhenae, radix scrophulariae, rehmanniae radix, cortex moutan, trichosanthis radix, fructus forsythiae, fructus Gardeniae, cortex Phellodendri, scutellariae radix, and radix Paeoniae Rubra.
At present, content detection items in the stomach-clearing coptis tablet standard only use berberine hydrochloride as an index, and geniposide, baicalin and sarsasapogenin (b a qi 257) are qualitatively identified by thin-layer chromatography, so that the method is too simple, the content of active ingredients of other medicinal materials is difficult to effectively reflect, and the quality control in the production process is not facilitated. Therefore, the method for separating and measuring the effective components of the Chinese medicinal composition for clearing stomach has important significance in the aspects of process control, product effectiveness and stability judgment and the like in production.
In the existing HPLC content detection method, only the effective components of two medicinal materials of coptis chinensis and phellodendron amurense of a product can be quantitatively detected by a single wavelength, and four corresponding effective components can be respectively detected by four wavelengths, so that the effective components of various other medicinal materials cannot be quantitatively controlled conveniently and comprehensively. The content detection of the coptis tablet for clearing stomach in the prior literature comprises the following steps: (1) geniposide, baicalin, berberine hydrochloride and ammonium glycyrrhizinate are used as indicators (the multi-wavelength HPLC method is used for simultaneously measuring the content of 4 components in the eclipta alba tablet, the Tangchun, the Lushimei, the Chinese herbal medicine, the 2014 06); (2) taking berberine hydrochloride, prasudine hydrochloride and palmatine hydrochloride which are related components of coptis as indicators (determination of content of berberine hydrochloride, prasudine hydrochloride and palmatine in the stomach-clearing rhizoma coptidis tablet, huyayan, tianjian, guangzhou chemical industry, 16 years 2012). However, the determination method still needs to be further improved, and the effective components to be determined need to be further improved.
Therefore, there is an urgent need to develop a method for detecting or separating the components of a QINGWEIHUANG tablet, which can detect more components and is stable, effective, simple and low in cost.
Disclosure of Invention
The invention aims to overcome the defects of a method for detecting or separating a component of a QINGWEIHUANG tablet in the prior art, and provides a method for detecting or separating a component of a QINGWEIHUANG tablet.
In order to solve the technical problems, the invention provides a method for detecting or separating components of a coptis tablet for clearing stomach, which comprises the step of detecting a test solution containing the coptis tablet for clearing stomach by using high performance liquid chromatography;
in the high performance liquid chromatography, the adopted mobile phase A is acetonitrile, and the mobile phase B is (volume concentration is) 0.09-0.11% (v/v) of phosphoric acid aqueous solution; gradient elution was performed using the mobile phase a and the mobile phase B under the following conditions:
| |
0 | 5 | 10 | 30 | 45 | 50 | 53 | 58 | 63 |
| Mobile phase A% | 10 | 10 | 15 | 25 | 40 | 90 | 90 | 10 | 10 |
| Mobile phase B% | 90 | 90 | 85 | 75 | 60 | 10 | 10 | 90 | 90 |
Wherein, the percentage in the table is the volume percentage of each mobile phase in the total volume of the mobile phase A and the mobile phase B respectively.
Preferably, the mobile phase B is 0.1% phosphoric acid aqueous solution.
Preferably, in the high performance liquid chromatography detection, the chromatographic column is an octadecylsilane chemically bonded silica chromatographic column, such as YMC-Triant C18; the specification of the octadecylsilane bonded silica gel chromatographic column is preferably 4.6mm × 250mm,5 μm,12nm.
Preferably, in the high performance liquid chromatography detection, the column temperature of the chromatographic column is 20-25 ℃, such as 21, 22, 23, 24, 25 ℃;
preferably, the detection wavelength of the high performance liquid chromatography is 200-400 nm, preferably 205nm;
preferably, the sample injection volume of the high performance liquid chromatography is 5-20 μ L, and preferably 10 μ L.
Preferably, the flow rate of the mobile phase is between 0.9 and 1.1ml/min, preferably 1ml/min.
Preferably, the solvent in the test solution is a solution consisting of methanol and hydrochloric acid, the methanol is preferably 100% methanol, and the volume of the methanol and the hydrochloric acid is preferably (99-100): 1.
Preferably, in the test solution, the volume ratio of the qingweihuanglian tablet to the solvent is 8mg/ml.
Preferably, the method further comprises the steps of preparing a reference solution and carrying out the high performance liquid chromatography detection on the reference solution so as to determine the attribution of each peak in the test solution according to the retention time of the reference solution; the reference substance solution comprises jasminoidin reference substance, phellodendrine hydrochloride reference substance, phillyrin A reference substance, palmatine hydrochloride reference substance, berberine hydrochloride reference substance and/or baicalin reference substance.
In a preferred embodiment, the method comprises the steps of:
(1) Preparing a test solution: 100% methanol was used: hydrochloric acid =100, dissolving the coptis chinensis tablet;
(2) Carrying out high performance liquid detection: adopting a YMC-Triant C18 chromatographic column; the mobile phase is as follows: acetonitrile-0.1% phosphoric acid aqueous solution, column temperature: 25 ℃, flow rate: 1.0mL/min, and the detection wavelength is 205nm;
the gradient elution procedure was as follows:
| time (min) | 0 | 5 | 10 | 30 | 45 | 50 | 53 | 58 | 63 |
| Mobile phase A% | 10 | 10 | 15 | 25 | 40 | 90 | 90 | 10 | 10 |
| Mobile phase B% | 90 | 90 | 85 | 75 | 60 | 10 | 10 | 90 | 90 |
Wherein, the percentage in the table is the volume percentage of each mobile phase in the total volume of the mobile phase.
Preferably:
the step (1) also comprises the steps of standing to room temperature after 30min of ultrasound, constant volume and shaking up; and/or the specification of the YMC-Triant C18 chromatographic column is 4.6mm multiplied by 250mm,5 mu m and 12nm; and/or, the method also comprises the steps of preparing a reference substance solution and carrying out the high performance liquid chromatography detection on the reference substance solution, wherein the reference substance solution comprises a geniposide reference substance, a phellodendrine hydrochloride reference substance, a phillyrin A reference substance, a palmatine hydrochloride reference substance, a berberine hydrochloride reference substance and a baicalin reference substance, and the attribution of each peak in the test substance solution is determined according to the retention time of the reference substance solution. In a preferred embodiment, the HPLC is performed using an LC-2030C HPLC from Shimadzu.
On the basis of the common knowledge in the field, the above preferred conditions can be combined randomly to obtain the preferred embodiments of the invention.
The reagents and starting materials used in the present invention are commercially available.
The positive progress effects of the invention are as follows:
the method for detecting or separating the components of the stomach-clearing coptis tablet can simultaneously measure the content of 6 effective components such as geniposide, phellodendrine hydrochloride, paeoniflorin, forsythoside A, baicalin, berberine hydrochloride and the like, and can simply, quickly, effectively, accurately and reliably separate and detect various effective components in the stomach-clearing coptis tablet, so that the quality of the stomach-clearing coptis tablet is controlled, the method has good repeatability, the recovery rate meets the requirements, the cost is lower, and the method is more favorable for industrial application.
Drawings
FIG. 1 is a graph showing the results of elution performed in example 2 with acetonitrile-0.1% acetic acid as a mobile phase and with a gradient of the method shown in Table 3.
FIG. 2 is a graph showing the results of elution performed with the mobile phase of acetonitrile-0.1% acetic acid water in example 2 and using the gradient of the method shown in Table 4.
FIG. 3 is a graph showing the results of elution performed with the mobile phase of acetonitrile-0.1% phosphoric acid aqueous solution in example 2 by the gradient method shown in Table 5.
FIG. 4 is a graph showing the results of elution performed in example 2 using acetonitrile-0.1% phosphoric acid aqueous solution as a mobile phase and a gradient of the method shown in Table 6.
FIG. 5 is a graph showing the results of elution performed with the mobile phase of acetonitrile-0.1% phosphoric acid aqueous solution in example 2 by the gradient method shown in Table 7.
FIG. 6 is a graph showing the results of elution performed with the mobile phase of acetonitrile-0.1% phosphoric acid aqueous solution in example 2 by the gradient method shown in Table 8.
FIG. 7 is a graph showing the results of elution performed at 20 ℃,25 ℃ and 30 ℃ in example 2.
FIGS. 8 and 15 show the profiles detected in example 3.
FIG. 9 is a jasminoidin standard curve.
FIG. 10 is a standard curve of phellodendrine hydrochloride.
Fig. 11 is a forsythoside a standard curve.
Fig. 12 is a palmatine hydrochloride standard curve.
FIG. 13 is a standard curve of berberine hydrochloride.
FIG. 14 is a baicalin standard curve.
Detailed Description
The invention is further illustrated by the following examples, which are not intended to limit the invention thereto. The experimental methods without specifying specific conditions in the following examples were selected according to the conventional methods and conditions, or according to the commercial instructions.
The samples of QINGWEIHUANGLIAN tablet used in the following examples were purchased from Wuhan Kangle pharmaceutical Co., ltd. Samples are produced from 2019 to 2021, and the national standard of medicine is Z20020080.
Example 1 target Peak determination
According to the comparative analysis of the bulk drugs, the medicinal material extracts and the finished product preparations, selecting an elution peak with obvious ultraviolet response under the HPLC condition, and determining the following target peaks: geniposide, phellodendrine hydrochloride, forsythoside A, palmatine hydrochloride, berberine hydrochloride and baicalin. Wherein the geniposide is from fructus Gardeniae, the phellodendrine hydrochloride is from cortex Phellodendri, forsythiaside A is from fructus forsythiae, palmatine hydrochloride is from Coptidis rhizoma, berberine hydrochloride is from Coptidis rhizoma and cortex Phellodendri, and baicalin is from Scutellariae radix.
Example 2 determination of chromatographic conditions
2.1 selection of sample solvents
1) Taking 0.40g of stomach-clearing coptis chinensis tablet powder, precisely weighing, placing in a 50mL volumetric flask, adding about 40mL of sample solvent, carrying out ultrasonic treatment to fully dissolve the sample solvent, standing to room temperature, metering to a certain volume, and shaking up to obtain the stomach-clearing coptis chinensis tablet. The sample solvents formulated are shown in table 1:
TABLE 1
| (1) 30% ethanol | (2) 50% ethanol | (3) 80% ethanol |
| (4) 30% methanol | (5) 50% methanol | (6) 80% methanol |
| (7) 30% acetonitrile | (8) 50% acetonitrile | (9) 80% acetonitrile |
As a result, it was found that the dissolution effect was the best in the three solvents (5), (6) and (8), but the flocculent precipitate at the bottom of the bottle was too much, and thus it was not adopted.
2) Taking 0.40g of clear stomach-clearing rhizoma coptidis tablet clear paste powder, precisely weighing, placing in a 50mL volumetric flask, adding about 40mL of sample solvent, performing ultrasonic treatment to fully dissolve the clear stomach-clearing rhizoma coptidis tablet clear paste powder, standing to room temperature, fixing the volume to the scale, and shaking up to obtain the finished product. The sample solvents formulated are shown in table 2:
TABLE 2
| (1) 50% methanol (1% hydrochloric acid) | (2) 80% methanol (1% hydrochloric acid) |
| (3) 100% methanol (1% hydrochloric acid) | (4) 50% acetonitrile (1% hydrochloric acid) |
As a result, it was found that the dissolution effect was optimized by the solvent (3), and the floc precipitation at the bottom of the bottle was very small, and this solvent was feasible.
2.2 selection of chromatographic column and Mobile phase System
1) The mobile phase was eluted with acetonitrile-0.1% acetic acid water in the gradient of the method in table 3: the separation effect is very poor, as shown in figure 1;
TABLE 3
2) The mobile phase was eluted with acetonitrile-0.1% acetic acid water using the gradient of the method in table 4: the analysis was very poor, see FIG. 2. Therefore, acetonitrile-0.1% phosphoric acid is selected as the mobile phase system.
TABLE 4
The above compares the peak appearance of acetonitrile-0.1% acetic acid solution under 2 kinds of gradients, and the baseline shift is found to be larger, so acetonitrile-0.1% acetic acid solution is not used.
2.3 selection of wavelength
And when the full-wavelength scanning graph of the 6 target peaks is combined, the wavelength of 205nm can simultaneously ensure that the 6 target peaks have higher absorption.
2.4 optimization of gradient method
Adopting YMC-Triant C18 (4.6 mm × 250mm,5 μm,12 nm) chromatographic column; the mobile phase is as follows: acetonitrile-0.1% phosphoric acid aqueous solution, column temperature: 25 ℃, flow rate: 1.0mL/min, and the detection wavelength is 205nm. The gradient methods and their results are as follows:
the method comprises the following steps: the gradient method in table 5 was used for detection, and the results are shown in fig. 3: phellodendrine hydrochloride and forsythiaside A are not separated.
TABLE 5
| Time (min) | 0 | 5 | 50 | 53 | 56 | 63 |
| Acetonitrile (%) | 10 | 10 | 55 | 90 | 10 | 10 |
| 0.1% phosphoric acid (%) | 90 | 90 | 85 | 10 | 90 | 90 |
The method 2 comprises the following steps: the gradient method in Table 6 was used for detection, and the results are shown in FIG. 4: the purity chart shows that the palmatine hydrochloride peak contains a foreign peak.
TABLE 6
The method 3 comprises the following steps: the gradient method in table 7 was used for detection, and the results are shown in fig. 5: the purity chart shows that the palmatine hydrochloride peak contains a hybrid peak.
TABLE 7
The method 4 comprises the following steps: the gradient method in Table 8 was used for detection, and the results are shown in FIG. 6:6 target peaks can be separated, and the method is feasible.
TABLE 8
2.5 selection of column temperature
Comparing the column temperatures of 20 deg.C, 25 deg.C and 30 deg.C, the separation effect of the sample was best at 20 deg.C and 25 deg.C, as shown in FIG. 7.
EXAMPLE 3 sample testing
1. Preparation of a test solution: taking 0.4mg of stomach-clearing coptis chinensis tablet powder, accurately weighing, placing in a 50mL volumetric flask, adding 100% methanol: hydrochloric acid =100 (v/v), shaking up for 30min, standing to room temperature, fixing the volume to the scale, and shaking up to obtain the final product.
2. Preparation of control solutions: taking a proper amount of geniposide, phellodendrine hydrochloride, forsythin A, palmatine hydrochloride, berberine hydrochloride and baicalin reference substances (all the reference substances are from China pharmaceutical and biological product inspection institute, the geniposide batch number is 110749-201919 content is 97.1%, the phellodendrine hydrochloride batch number is 111895-201805 content is 94.9%, the forsythin A batch number is 111810-201707 content is 97.2%, the palmatine hydrochloride batch number is 110732-201611 content is 86.8%, the berberine hydrochloride batch number is 110713-201814 content is 86.7%, and the baicalin batch number is 110715-201821 content is 95.4%), precisely weighing, placing the precisely weighed materials in a 50mL volumetric flask, adding 100% methanol: hydrochloric acid =100 (v/v), 1 (v/v), about 40ml of mixed solution, shaking up, performing ultrasonic treatment for 30min, standing to room temperature, fixing the volume to scale, and shaking up to obtain the mixed control solution, wherein the mixed control solution is prepared from 0.1mg/ml of geniposide, 0.1mg/ml of berberine hydrochloride, 0.1mg/ml of baicalin, 0.01mg/ml of phellodendrine hydrochloride, 0.01mg/ml of forsythoside A and 0.01mg/ml of palmatine hydrochloride.
3. Setting high performance liquid detection conditions: adopting YMC-Triant C18 (4.6 mm × 250mm,5 μm,12 nm) chromatographic column; the mobile phase is as follows: acetonitrile-0.1% phosphoric acid aqueous solution, column temperature: 25 ℃, flow rate: 1.0mL/min, and the detection wavelength is 205nm. The number of theoretical plates is not less than 60000 calculated by berberine hydrochloride peak, jasminoidin is not less than 40000, phellodendrine hydrochloride is not less than 40000, phillyrin A is not less than 50000, palmatine hydrochloride is not less than 200000, and baicalin is not less than 300000.
The gradient elution procedure is shown in table 9 below.
TABLE 9
| |
0 | 5 | 10 | 30 | 45 | 50 | 53 | 58 | 63 |
| 0.1% phosphoric acid solution (%) | 90 | 90 | 85 | 75 | 60 | 10 | 10 | 90 | 90 |
| Acetonitrile (%) | 10 | 10 | 15 | 25 | 40 | 90 | 90 | 10 | 10 |
4. Precisely sucking 10 μ L of each of the reference solution and the sample solution, injecting into a liquid chromatograph, and recording chromatogram.
The results are shown in FIGS. 8 and 15.
In fig. 8, the degrees of separation are good for geniposide No. 1 (t = 16.121), phellodendrine hydrochloride No. 2 (t = 16.88), phillyrin a No. 3 (t = 25.203), palmatine hydrochloride No. 4 (t = 40.445), berberine hydrochloride No. 5 (t = 41.515), and baicalin No. 6 (t = 43.449). B-D, which are the enlarged isolated views of the corresponding maps in the boxes from left to right in a, respectively, wherein B is mazetasin 1 (t = 16.121), mazetasin hydrochloride 2 (t = 16.88), C is mazetasin 3 (t = 25.203), D is mazetasin hydrochloride 4 (t = 40.445), mazetasin hydrochloride 5 (t = 41.515), and mazetasin 6 (t = 43.449), and the isolation is good.
Example 4 method verification of stomach-clearing Coptis chinensis tablet
1) Apparatus and materials
1.1 Experimental instruments
ME204 ten thousandth analytical balance (METTLER TOLEDO);
a synergy ultra-pure water machine (MERCK MILLIPORE);
KUDOS SK7200HP model ultrasonic cleaner (shanghai kokai ultrasonic instruments ltd);
LC-2030C high performance liquid chromatograph (Shimadzu);
1.2 Experimental reagents
Isopropyl alcohol (analytical pure AR, national chemical group, ltd);
phosphoric acid (chromatographic grade, camey chemie ltd, tianjin);
methanol (chromatographic grade, MREDA);
acetonitrile (chromatographic grade, MREDA);
hydrochloric acid (analytically pure AR, chemical reagents of national medicine group Co., ltd.)
Ultrapure water (self-made)
Geniposide (Chinese medicine biological product inspection institute, batch number: 110749-201919, content 97.1%)
Phellodendrine hydrochloride (Chinese medicine biological product inspection institute, batch number: 111895-201805, content 94.9%)
Forsythoside A (China pharmaceutical and biological products institute, batch number: 111810-201707, content 97.2%)
Palmatine hydrochloride (China pharmaceutical and biological products institute, batch number: 110732-201611, content 86.8%)
Berberine hydrochloride (China pharmaceutical biological products institute, batch number: 110713-201814, content 86.7%)
Baicalin (Chinese medicine biological product inspection institute, batch number: 110715-201821, content 95.4%)
1.3 preparation of solution
0.1% phosphoric acid aqueous solution: measuring phosphoric acid in a volumetric flask of 1mL to 1000mL, metering the volume to the scale with ultrapure water, shaking up, performing suction filtration, filtering through a 0.45 mu m filter membrane, and performing ultrasonic treatment for 5min to obtain the product;
acetonitrile: weighing 1000mL of acetonitrile, performing suction filtration, filtering through a 0.45-micron filter membrane, and performing ultrasonic treatment for 5min to obtain the product;
methanol: weighing 1000mL of methanol, performing suction filtration, filtering with 0.45 μm filter membrane, and performing ultrasonic treatment for 5min to obtain the final product;
10% isopropyl alcohol: respectively weighing 100mL of isopropanol and 900mL of ultrapure water, mixing, shaking, filtering with a 0.45-micrometer filter membrane, and performing ultrasonic treatment for 5min to obtain the final product;
10% of methanol: respectively weighing 100mL of methanol and 900mL of ultrapure water, mixing, shaking, performing suction filtration, filtering through a 0.45-micrometer filter membrane, and performing ultrasonic treatment for 5min to obtain the product;
50% methanol: respectively weighing 500mL of methanol and 500mL of ultrapure water, mixing, shaking, performing suction filtration, filtering through a 0.45-micrometer filter membrane, and performing ultrasonic treatment for 5min to obtain the product;
1% hydrochloric acid methanol (solvent): measuring hydrochloric acid 10mL to 1000mL volumetric flask, metering to a certain volume with ultrapure water to scale, shaking up, and performing ultrasonic treatment for 5min to obtain the product;
2) Experimental methods
2.1 preparation of test solutions
Weighing 0.40g of sample (QINGWEIHUANGLIAN tablet powder), placing in a 50mL volumetric flask, adding about 40mL of 1% methanol hydrochloride, performing ultrasonic treatment to fully dissolve the sample, standing to room temperature, fixing the volume to the scale, shaking up, filtering with a 0.45 μm microporous membrane, and taking the subsequent filtrate to obtain the sample solution.
2.2 preparation of control solutions
Precisely weighing each reference substance, placing in a 50mL volumetric flask, adding about 30mL of 1% methanol hydrochloride, ultrasonically dissolving, standing to room temperature, fixing the volume to the scale, shaking uniformly, and preparing into a mixed reference solution of 0.1mg/mL of geniposide, 0.1mg/mL of berberine hydrochloride, 0.1mg/mL of baicalin, 0.01mg/mL of phellodendrine hydrochloride, 0.01mg/mL of forsythin and 0.01mg/mL of palmatine hydrochloride.
2.3 chromatographic conditions
Adopting YMC-Triant C18 (4.6 mm × 250mm,5 μm,12 nm) chromatographic column; the mobile phase is as follows: acetonitrile-0.1% phosphoric acid aqueous solution, column temperature: 25 ℃, flow rate: 1.0mL/min, and the detection wavelength is 205nm. Gradient elution was performed as specified in table 10:
watch 10
| Time (min) | 0 | 5 | 10 | 30 | 45 | 50 | 53 | 58 | 63 |
| Acetonitrile (%) | 10 | 10 | 15 | 25 | 40 | 90 | 90 | 10 | 10 |
| 0.1% phosphoric acid (%) | 90 | 90 | 85 | 75 | 61 | 10 | 10 | 90 | 90 |
3. Verification experiment
3.1 Linear Range inspection
The control solutions of each control were precisely prepared as follows. Sample introduction is carried out according to the chromatographic conditions under the item of 2.3 respectively, and the chromatographic peak area is recorded. Linear regression analysis was performed with the concentration x (mg/mL) as the abscissa and the peak area y (mAU. Min) as the ordinate. The results are shown in FIGS. 9-14.
Preparation of control solutions
Geniposide-1 control: precisely weighing 8.04mg of geniposide, placing in a 25mL volumetric flask, adding about 15mL of 1% methanol hydrochloride, performing ultrasonic treatment to fully dissolve the geniposide, standing to room temperature, fixing the volume to scale, and shaking up to obtain the gardenoside;
geniposide-2 control: gardenoside-1 control solution 12.5mL +1% methanol hydrochloride 12.5mL;
geniposide-3 control: gardenoside-2 control solution 12.5mL +1% methanol hydrochloride 37.5mL;
geniposide-4 control: geniposide-3 control liquid 10.0mL +1% methanol hydrochloride 40.0mL;
geniposide-5 control solution: jasminoidin-4 control solution 10.0mL +1% methanol hydrochloride 90.0mL;
phellodendrine hydrochloride-1 control solution: taking 5.02mg of phellodendrine hydrochloride, precisely weighing, placing in a 50mL volumetric flask, adding about 30mL of 1% methanol hydrochloride, performing ultrasonic treatment to fully dissolve the phellodendrine hydrochloride, standing to room temperature, metering volume to a scale, and shaking up to obtain the phellodendrine hydrochloride scale;
phellodendrine hydrochloride-2 control solution: phellodendrine hydrochloride-1 reference solution 5mL of 20mL of 1% methanol hydrochloride;
phellodendrine hydrochloride-3 control solution: phellodendrine hydrochloride-2 reference solution 12.5mL +1% methanol hydrochloride 12.5mL;
phellodendrine hydrochloride-4 control solution: phellodendrine hydrochloride-3 reference solution 10.0mL +1% methanol hydrochloride 90.0mL;
phellodendrine hydrochloride-5 control solution: phellodendrine hydrochloride-4 reference solution 10.0mL +1% methanol hydrochloride 40.0mL;
forsythoside A-1 control solution: taking 5.17mg of forsythiaside A as a reference substance, precisely weighing, placing in a 50mL volumetric flask, adding about 30mL of 1% methanol hydrochloride, performing ultrasonic treatment to fully dissolve, standing to room temperature, fixing the volume to a scale, and shaking up to obtain the forsythiaside A;
forsythoside A-2 control solution: forsythoside A-1 control liquid 5mL +1% hydrochloric acid methanol 20mL;
forsythoside A-3 control solution: forsythoside A-2 control solution 12.5mL +1% methanol hydrochloride 12.5mL;
forsythoside A-4 control solution: forsythoside A-3 control solution 10.0mL +1% methanol hydrochloride 90.0mL;
forsythoside A-5 control solution: forsythoside A-4 control solution 10.0mL +1% methanol hydrochloride 40.0mL;
palmatine hydrochloride-1 control solution: precisely weighing 4.80mg of palmatine hydrochloride, placing in a 50mL volumetric flask, adding about 30mL of 1% methanol hydrochloride, performing ultrasonic treatment to fully dissolve the palmatine hydrochloride, standing to room temperature, fixing the volume to scale, and shaking up to obtain the palmatine hydrochloride tablet;
palmatine hydrochloride-2 control solution: bamagastine hydrochloride-1 reference liquid 5mL +1% methanol hydrochloride 20mL;
palmatine hydrochloride-3 control solution: palmatine hydrochloride-2 reference solution 12.5mL +1% methanol hydrochloride 12.5mL;
palmatine hydrochloride-4 control solution: palmatine hydrochloride-3 control solution 10.0mL +1% methanol hydrochloride 90.0mL;
palmatine hydrochloride-5 control solution: palmatine hydrochloride-4 control solution 10.0mL +1% methanol hydrochloride 40.0mL;
note: phellodendrine hydrochloride, forsythoside A and palmatine hydrochloride are three kinds of control mixed liquid.
Berberine hydrochloride-1 control solution: taking 8.08mg of berberine hydrochloride, precisely weighing, placing in a 25mL volumetric flask, adding about 15mL of 1% methanol hydrochloride, performing ultrasonic treatment to fully dissolve the berberine hydrochloride, standing to room temperature, fixing the volume to scale, and shaking up to obtain the berberine hydrochloride;
berberine hydrochloride-2 reference solution: berberine hydrochloride-1 reference solution 12.5mL +1% methanol hydrochloride 12.5mL;
berberine hydrochloride-3 reference solution: berberine hydrochloride-2 reference solution 12.5mL +1% methanol hydrochloride 37.5mL;
berberine hydrochloride-4 reference solution: berberine hydrochloride-3 contrast solution 10.0mL +1% methanol hydrochloride 40.0mL;
berberine hydrochloride-5 reference solution: berberine hydrochloride-4 contrast solution 10.0mL +1% methanol hydrochloride 90.0mL;
baicalin-1 control solution: precisely weighing 9.78mg of baicalin, placing in a 10mL volumetric flask, adding about 7mL of 1% methanol hydrochloride, performing ultrasonic treatment to fully dissolve the baicalin, standing to room temperature, fixing the volume to scale, and shaking up to obtain the product;
baicalin-2 control solution: baicalin-1 control solution 4mL of 1% methanol hydrochloride 6mL;
baicalin-3 control solution: baicalin-2 control solution 5mL of 1% methanol hydrochloride 5mL;
baicalin-4 control solution: baicalin-3 control liquid, 45mL of 1% methanol hydrochloride and 5mL;
baicalin-5 control solution: baicalin-4 control liquid 10mL +1% methanol hydrochloride 90mL;
as a result, the
Linear regression analysis was performed with the concentration x (g/mL) as the abscissa and the peak area y (mAU. Min) as the ordinate. The linear regression equations and correlation coefficients for the six controls are shown in FIGS. 9-14 below.
3.2 precision test
Precisely sucking 10 μ L of QINGWEIHUANGLIAN tablet sample solution (respectively labeled as precision samples-1-6), repeatedly injecting sample under the condition of chromatography of "2.3" for 6 times, and measuring peak area. The peak areas and RSD values of the six reference substances are shown in Table 11, the RSD% of the reference substances is less than 2%, and the results show that the precision of the instrument method is good.
TABLE 11 results of precision tests
3.3 stability test
Precisely sucking 10 mu L of the sample solution of the stomach-clearing coptis tablet, respectively injecting samples (respectively marked as stability samples of 0h, 2h, 8h, 12h and 24 h) at the chromatographic conditions of '2.3', and determining peak areas. The content of the index substances in each time period and the RSD value thereof are shown in a table 12, the RSD% of the index substances is less than 2%, and the result shows that the precision of the instrument method is good.
Table 12 stability test results
3.4 repeatability tests
Taking 6 parts of the same batch of stomach-clearing coptis chinensis tablet samples (respectively marked as repetitive samples-1 to 6), preparing a sample solution according to the method under the item '2.1', respectively injecting samples according to the chromatographic condition under the item '2.3', measuring the retention time and peak area of six indexes, calculating the content of the six indexes by an external standard method, and calculating the RSD% value of the six indexes. The RSD% of the method meets the requirements within the limit range of the content of each component, the result shows that the repeatability of the determination method is good, and the repeatability test results are shown in tables 13 and 14.
TABLE 13 retention time test results
TABLE 14 content test results
3.5 sample application recovery test
Precisely weighing 6 parts of the same batch of stomach-clearing coptis chinensis tablet samples with known content, adding an equal-content mixed reference substance, preparing a sample solution according to the method under the item '2.1', respectively injecting samples according to the chromatographic condition under the item '2.3', measuring the retention time and peak area of six indexes, and calculating the content by an external standard method. The results show that the recovery rate of the determination method meets the requirements, and the test results are shown in Table 15.
3.6 limit of quantitation
TABLE 16
3.7 detection Limit
TABLE 17
| Name of index | Gardenia glycoside | Phellodendrine hydrochloride | Forsythoside A |
| Concentration (g/mL) | 2.6×10 -07 | 6.3×10 -08 | 6.7×10 -08 |
| Name of index | Palmatine hydrochloride | Berberine hydrochloride | Baicalin |
| Concentration (g/mL) | 5.7×10 -08 | 2.3×10 -07 | 6.3×10 -07 |
Claims (10)
1. A method for detecting or separating components of a Chinese medicinal composition for clearing stomach comprises detecting a test solution containing the Chinese medicinal composition for clearing stomach by high performance liquid chromatography;
in the high performance liquid chromatography, the adopted mobile phase A is acetonitrile, and the mobile phase B is 0.09-0.11% phosphoric acid aqueous solution; gradient elution was performed using the mobile phase a and the mobile phase B under the following conditions:
Wherein, the percentage in the table is the volume percentage of each mobile phase in the total volume of the mobile phases.
2. The method of claim 1, wherein the mobile phase B is a 0.1% aqueous phosphoric acid solution.
3. The method of claim 1 or 2, wherein the high performance liquid chromatography is performed using an octadecylsilane-bonded silica gel column, such as YMC-Triant C18.
4. The method of claim 3, wherein the octadecylsilane bonded silica chromatography column has a specification of 4.6mm x 250mm,5 μm,12nm.
5. The method according to any one of claims 1 to 4, wherein the column temperature of the chromatographic column in the high performance liquid chromatography detection is 20 to 25 ℃;
and/or the detection wavelength of the high performance liquid chromatography is 200-400 nm, preferably 205nm;
and/or the sample injection volume of the high performance liquid chromatography is 5-20 mu L, preferably 10 mu L.
6. The method according to any one of claims 1 to 5, wherein the flow rate of the mobile phase is between 0.9 and 1.1ml/min, preferably 1ml/min;
and/or, in the test solution, the volume ratio of the QINGWEIHUANG tablet to the solvent is 8mg/ml.
7. The method of claim 6, wherein the solvent in the test solution is a solution of methanol and hydrochloric acid, wherein the methanol is preferably 100% methanol, and the volume of the methanol and the hydrochloric acid is preferably (99-100): 1.
8. The method of claims 1-7, further comprising the steps of formulating a control solution and subjecting the control solution to the high performance liquid chromatography assay to determine the assignment of peaks in the test solution based on the retention time of the control solution; the reference substance solution comprises jasminoidin reference substance, phellodendrine hydrochloride reference substance, phillyrin A reference substance, palmatine hydrochloride reference substance, berberine hydrochloride reference substance and/or baicalin reference substance.
9. The method according to any one of claims 1 to 8, characterized in that it comprises the steps of:
(1) Preparation of a test solution: 100% methanol was used: hydrochloric acid =100, dissolving the coptis chinensis tablet;
(2) Carrying out high performance liquid detection: adopting a YMC-Triant C18 chromatographic column; the mobile phase is as follows: acetonitrile-0.1% phosphoric acid aqueous solution, column temperature: 25 ℃, flow rate: 1.0mL/min, and the detection wavelength is 205nm;
the gradient elution procedure was as follows:
wherein, the percentage in the table is the volume percentage of each mobile phase in the total volume of the mobile phases.
10. The method of claim 9, wherein the step (1) preferably further comprises the steps of standing to room temperature after 30min of ultrasound treatment, constant volume and shaking up; and/or, the size of the YMC-Triant C18 chromatographic column is preferably 4.6mm multiplied by 250mm,5 mu m and 12nm.
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|---|---|---|---|---|
| CN115343377A (en) * | 2021-05-14 | 2022-11-15 | 武汉康乐药业股份有限公司 | Fingerprint spectrum of stomach-clearing coptis tablet and construction method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102068673A (en) * | 2009-11-20 | 2011-05-25 | 天津中新药业集团股份有限公司乐仁堂制药厂 | Quality control method of stomach-strengthening and chest-relieving pill as Chinese herbal preparation |
| CN102707009A (en) * | 2012-06-18 | 2012-10-03 | 天津中新药业集团股份有限公司达仁堂制药厂 | Method for detecting rhizoma coptidis pills for clearing away heat of upper part of body |
| CN115343377A (en) * | 2021-05-14 | 2022-11-15 | 武汉康乐药业股份有限公司 | Fingerprint spectrum of stomach-clearing coptis tablet and construction method and application thereof |
-
2021
- 2021-05-14 CN CN202110529261.2A patent/CN115343376A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102068673A (en) * | 2009-11-20 | 2011-05-25 | 天津中新药业集团股份有限公司乐仁堂制药厂 | Quality control method of stomach-strengthening and chest-relieving pill as Chinese herbal preparation |
| CN102707009A (en) * | 2012-06-18 | 2012-10-03 | 天津中新药业集团股份有限公司达仁堂制药厂 | Method for detecting rhizoma coptidis pills for clearing away heat of upper part of body |
| CN115343377A (en) * | 2021-05-14 | 2022-11-15 | 武汉康乐药业股份有限公司 | Fingerprint spectrum of stomach-clearing coptis tablet and construction method and application thereof |
Non-Patent Citations (4)
| Title |
|---|
| 国家药典委员会: "中国药典2010年版一部", 31 December 2010, 中国医药科技出版社, pages: 63 - 64 * |
| 孙承浩等: "HPLC同时分析芎菊上清丸中7个化学成分的含量研究", 中国生化药物杂志, vol. 34, no. 7, 31 December 2014 (2014-12-31), pages 177 - 183 * |
| 王静竹等: "三种中成药中小檗碱和巴马汀的含量测定", 中国实验方剂学杂志, vol. 3, no. 1, 31 December 1997 (1997-12-31), pages 2 - 3 * |
| 高岩等: "基于多成分同时定量清胃黄连丸质量研究", 中药材, vol. 43, no. 8, 31 August 2020 (2020-08-31), pages 1946 - 1951 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115343377A (en) * | 2021-05-14 | 2022-11-15 | 武汉康乐药业股份有限公司 | Fingerprint spectrum of stomach-clearing coptis tablet and construction method and application thereof |
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