CN114252532B - Method for establishing fingerprint of Fangfengsheng particles and fingerprint thereof - Google Patents
Method for establishing fingerprint of Fangfengsheng particles and fingerprint thereof Download PDFInfo
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- 239000004378 Glycyrrhizin Substances 0.000 claims description 9
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- LPLVUJXQOOQHMX-UHFFFAOYSA-N glycyrrhetinic acid glycoside Natural products C1CC(C2C(C3(CCC4(C)CCC(C)(CC4C3=CC2=O)C(O)=O)C)(C)CC2)(C)C2C(C)(C)C1OC1OC(C(O)=O)C(O)C(O)C1OC1OC(C(O)=O)C(O)C(O)C1O LPLVUJXQOOQHMX-UHFFFAOYSA-N 0.000 claims description 9
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- UYRUBYNTXSDKQT-UHFFFAOYSA-N glycyrrhizic acid Natural products CC1(C)C(CCC2(C)C1CCC3(C)C2C(=O)C=C4C5CC(C)(CCC5(C)CCC34C)C(=O)O)OC6OC(C(O)C(O)C6OC7OC(O)C(O)C(O)C7C(=O)O)C(=O)O UYRUBYNTXSDKQT-UHFFFAOYSA-N 0.000 claims description 9
- 235000019410 glycyrrhizin Nutrition 0.000 claims description 9
- XSDQTOBWRPYKKA-UHFFFAOYSA-N amiloride Chemical compound NC(=N)NC(=O)C1=NC(Cl)=C(N)N=C1N XSDQTOBWRPYKKA-UHFFFAOYSA-N 0.000 claims description 8
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- ILRKKHJEINIICQ-OOFFSTKBSA-N Monoammonium glycyrrhizinate Chemical compound N.O([C@@H]1[C@@H](O)[C@H](O)[C@H](O[C@@H]1O[C@H]1CC[C@]2(C)[C@H]3C(=O)C=C4[C@@H]5C[C@](C)(CC[C@@]5(CC[C@@]4(C)[C@]3(C)CC[C@H]2C1(C)C)C)C(O)=O)C(O)=O)[C@@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O ILRKKHJEINIICQ-OOFFSTKBSA-N 0.000 claims description 2
- 239000001685 glycyrrhizic acid Substances 0.000 claims description 2
- 229940088594 vitamin Drugs 0.000 claims description 2
- 239000011782 vitamin Substances 0.000 claims description 2
- MQBFFYQCZCKSBX-UHFFFAOYSA-N 5-hydroxy-6,7,8-trimethoxy-2-(3,4,5-trimethoxyphenyl)chromen-4-one Chemical compound COC1=C(OC)C(OC)=CC(C=2OC3=C(OC)C(OC)=C(OC)C(O)=C3C(=O)C=2)=C1 MQBFFYQCZCKSBX-UHFFFAOYSA-N 0.000 claims 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 claims 1
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- 240000001972 Gardenia jasminoides Species 0.000 claims 1
- 229930188400 Gardenin Natural products 0.000 claims 1
- 239000012467 final product Substances 0.000 claims 1
- 229930182478 glucoside Natural products 0.000 claims 1
- 150000008131 glucosides Chemical class 0.000 claims 1
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- 235000013343 vitamin Nutrition 0.000 claims 1
- 150000003722 vitamin derivatives Chemical class 0.000 claims 1
- 238000004458 analytical method Methods 0.000 abstract description 2
- 239000000463 material Substances 0.000 abstract description 2
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 abstract description 2
- 238000003908 quality control method Methods 0.000 abstract description 2
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- FURUXTVZLHCCNA-UHFFFAOYSA-N Liquiritigenin Natural products C1=CC(O)=CC=C1C1OC2=CC(O)=CC=C2C(=O)C1 FURUXTVZLHCCNA-UHFFFAOYSA-N 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- JBQATDIMBVLPRB-UHFFFAOYSA-N isoliquiritigenin Natural products OC1=CC(O)=CC=C1C1OC2=CC(O)=CC=C2C(=O)C1 JBQATDIMBVLPRB-UHFFFAOYSA-N 0.000 description 2
- FURUXTVZLHCCNA-AWEZNQCLSA-N liquiritigenin Chemical compound C1=CC(O)=CC=C1[C@H]1OC2=CC(O)=CC=C2C(=O)C1 FURUXTVZLHCCNA-AWEZNQCLSA-N 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- GSZUGBAEBARHAW-UHFFFAOYSA-N sophoraflavone B Natural products OC1C(O)C(O)C(CO)OC1OC1=CC=C(C=2OC3=CC(O)=CC=C3C(=O)C=2)C=C1 GSZUGBAEBARHAW-UHFFFAOYSA-N 0.000 description 2
- 229940126680 traditional chinese medicines Drugs 0.000 description 2
- 201000004624 Dermatitis Diseases 0.000 description 1
- 241000218671 Ephedra Species 0.000 description 1
- 241000229182 Ledebouriella seseloides Species 0.000 description 1
- 235000006679 Mentha X verticillata Nutrition 0.000 description 1
- 235000002899 Mentha suaveolens Nutrition 0.000 description 1
- 235000001636 Mentha x rotundifolia Nutrition 0.000 description 1
- 206010057190 Respiratory tract infections Diseases 0.000 description 1
- 241000951473 Schizonepeta Species 0.000 description 1
- 206010046306 Upper respiratory tract infection Diseases 0.000 description 1
- 208000024780 Urticaria Diseases 0.000 description 1
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—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
- G01N30/62—Detectors specially adapted therefor
- G01N30/74—Optical detectors
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/86—Signal analysis
- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
- G01N30/8686—Fingerprinting, e.g. without prior knowledge of the sample components
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- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- General Physics & Mathematics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Pathology (AREA)
- General Health & Medical Sciences (AREA)
- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Engineering & Computer Science (AREA)
- Library & Information Science (AREA)
- Medicinal Preparation (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a method for establishing fingerprint of Fangfengtong sheng particles and the fingerprint thereof, belonging to the field of analysis of traditional Chinese medicine preparations. The fingerprint spectrum establishment method comprises the following steps: preparing a sample solution; preparing a mixed reference substance solution; measuring with high performance liquid chromatograph; and (3) constructing fingerprint of the Fengshong sheng particles. Chromatographic conditions: octadecylsilane chemically bonded silica gel is adopted as a filling material; the detection wavelength is 254nm; the flow rate is 1.0ml/min; the column temperature is 25 ℃; the sample injection amount is 10 μl; acetonitrile and 0.1% phosphoric acid aqueous solution were used as mobile phases for gradient elution. The method has the advantages of good repeatability, high stability and precision, simple operation and the like. The fingerprint of the Fangfengsheng particles established by the method has 22 common peaks, 7 common characteristic peaks are identified, and the baicalin peak of the No. 12 peak is selected as the reference peak in the fingerprint, so that the quality control method of the Fangfengsheng particles is perfected, the product quality can be comprehensively controlled on the whole, and the effectiveness and the safety of the Fangfengsheng particles are ensured.
Description
Technical field:
the invention relates to the field of traditional Chinese medicine preparation analysis, in particular to a method for establishing fingerprint of Fangfengsheng particles and the fingerprint thereof.
The background technology is as follows:
the Fengshengsheng granule is derived from the Xuanming theory prescription, is one of the classical representation of the four-well-known Liu-complete element of Jinyuan, is composed of seventeen traditional Chinese medicines such as radix sileris, fructus forsythiae, schizonepeta spike, mint, ephedra and the like, has the effects of clearing heat and detoxicating, relieving exterior syndrome and dredging interior, can be widely used for treating upper respiratory infection, eczema, urticaria and the like clinically, and has obvious clinical curative effect. The Fengshong sheng granule has large prescription and complex medicinal flavor, is loaded in the first part of the 2020 edition of Chinese pharmacopoeia, comprises 9 physicochemical identifications and 2 content determinations, can control the quality of medicines to a certain extent, but lacks integral control for Chinese patent medicines with complex components and multiple targets. The traditional Chinese medicine fingerprint with the characteristics of 'integrity' and 'ambiguity' is used for evaluating the quality of traditional Chinese medicine materials, semi-finished products and traditional Chinese medicine preparations, and can comprehensively reflect the internal quality of traditional Chinese medicine products.
The invention comprises the following steps:
the invention mainly aims to solve the problem that the existing Fengsheng granules lack integrity in quality control, and provides a method for establishing a fingerprint of Fengsheng granules. The method ensures the stability, consistency and controllability of the quality of the Fengsheng granules.
To achieve the above object, the present invention is achieved by:
the method for establishing the fingerprint of the Fangfengsheng granule is characterized by comprising the following steps of:
1. preparation of test solution: grinding radix Saposhnikoviae, sieving with a fifth sieve, precisely weighing, placing in a conical bottle with a plug, adding a certain amount of solvent, ultrasonic extracting, cooling, adding solvent, shaking, filtering, and filtering with microporous membrane.
2. Preparation of a mixed control solution: and (3) taking a reference substance of geniposide, cimicifuga rhizome glycoside, glycyrrhizin, 5-O-methylweisi amiloride, baicalin, ammonium glycyrrhizinate and aloe-emodin, precisely weighing, adding a proper amount of solvent for dissolving, and passing through a microporous filter membrane to obtain a mixed reference substance solution.
3. And (3) measuring: respectively precisely sucking the sample solution and the mixed reference solution, injecting into a high performance liquid chromatograph, measuring, and recording the chromatogram under the following chromatographic conditions:
chromatographic column: agilent ZORBAX Eclipse Plus C column (5 μm, 4.6X1250 mm); the detection wavelength is 254nm; the flow rate is 1.0ml/min; the column temperature is 25 ℃; the sample injection amount is 10 μl; mobile phase A is 0.1% phosphoric acid aqueous solution, mobile phase B is acetonitrile, and gradient elution is adopted.
4. Construction of fingerprint of Fengshong Sheng particles: introducing different batches of sample chromatograms into traditional Chinese medicine chromatographic fingerprint similarity evaluation software, selecting common chromatographic peaks in each batch of sample to generate a Fangtong sheng particle control fingerprint, and calculating the relative peak area and relative retention time of each common peak.
As a preferred scheme, the method for establishing fingerprint of the Fengshong sheng particles is characterized in that in the step 1, the preparation method of the sample solution is as follows: respectively taking 15 batches of Fangfengsheng particles, grinding about 2g of particles, sieving with a five-size sieve, precisely weighing, placing into a conical flask with a plug, precisely adding 25ml of 70% methanol, weighing, ultrasonically extracting for 30 minutes, cooling, weighing again, supplementing the weight of loss with 70% methanol, shaking uniformly, filtering, collecting the filtrate, and passing through a 0.45 μm microporous filter membrane.
As a preferred scheme, the method for establishing fingerprint of the Fengshong sheng particles is characterized in that in the step 2, the preparation method of the mixed reference substance solution is as follows: accurately weighing reference substances including geniposide, cimicifuga rhizome glycoside, glycyrrhizin, 5-O-methylvitamin amiloride, baicalin, ammonium glycyrrhizinate, aloe-emodin, dissolving in 70% methanol, and making into geniposide (50 μg ml) -1 ) Cimicifugal glycoside (20. Mu.g.ml) -1 ) Liquiritigenin (20 mug.ml) -1 ) 5-O-methyl-Weisi-amiloride (10. Mu.g.ml) -1 ) Baicalin (0.4 mg.mL) -1 ) Ammonium glycyrrhizinate (0.20 mg.ml) -1 ) Aloe-emodin (5 mug. Ml) -1 ) Is filtered through a microporous membrane of 0.45 μm.
As a preferred scheme, the method for establishing fingerprint of the Fengshong sheng particles is characterized in that in the step 3, the gradient elution method is as follows:
in the preferred scheme, the method for establishing the fingerprint of the Fengsheng granules is characterized in that in the step 4, the fingerprint obtained by the establishment method contains 22 characteristic peaks, wherein the No. 2 peak, the No. 3 peak, the No. 5 peak, the No. 9 peak, the No. 12 peak, the No. 19 peak and the No. 20 peak are respectively geniposide, cimicifuga rhizome glycoside, glycyrrhizin, 5-O-methyl Weis amiloride, baicalin, ammonium glycyrrhizinate and aloe-emodin. Calculating the relative retention time and relative peak area of other 21 characteristic peaks by taking baicalin of No. 12 peak as a reference peak: peak No. 1 peak 0.1902 ± 0.0003,2 peak 0.3131 ± 0.0014,3 peak 0.4669 ± 0.0019,4 peak 0.5874 ± 0.0020,5 peak 0.6102 ± 0.0020,6 peak 0.6426 ± 0.0019,7 peak 0.6565 ± 0.0017,8 peak 0.6747 ± 0.0016,9 peak 0.7332 ±0.0016, peak 10 0.9459 ±0.0003, peak 11 peak 0.9741 ±0.0003, peak 13 0.9741 ±0.0004, peak 14 peak 0.9741 ±0.0003, peak 15 peak 1.1720±0.0006, peak 16 peak 1.2308±0.0008, peak 17 peak 0.9741 ±0.0010, peak 18 peak 0.9741 ±0.0011, peak 19 peak 0.9741 ±0.0012, peak 20 peak 0.9741 ±0.0014, peak 21 peak 0.9741 ±0.0016, peak 22 peak 0.9741 ±0.0016; relative peak area: peak No. 1, peak No. 0.0581 ± 0.0002,2, peak No. 0.0581 ± 0.0010,3, peak No. 0.0414 ± 0.0007,5, peak No. 0.1306± 0.0003,6, peak No. 0.0373± 0.0005,7, peak No. 0.0232 ± 0.0004,8, peak No. 0.0523±0.0006, peak No. 10, 0427±0.0001, peak No. 11, peak No. 0.0527 ±0.0005, peak No. 13, peak No. 0.0571 ±0.0001, peak No. 14, peak No. 0318+0.0004, peak No. 15, peak No. 0.2252 ±0.0041, peak No. 0.3343 ±0.0006, peak No. 17, peak No. 0.0865 ±0.0002, peak No. 18, peak No. 0.1545 ±0.0002, peak No. 20, peak No. 0.0280±0.0001, peak No. 21, peak No. 0199±0.0002, and peak No. 22, peak No. 0.0248 ±0.0004.
By using the method to measure 15 batches of Fangtong sheng particle samples, 15 batches of Fangtong sheng particle fingerprints are obtained, and the similarity of the HPLC (high performance liquid chromatography) fingerprints of all batches of Fangtong sheng particles is evaluated by adopting 2012 edition of the similarity evaluation system of traditional Chinese medicine chromatographic fingerprints, wherein the similarity of 15 batches of Fangtong sheng particles is larger than 0.99, which indicates that the quality of Fangtong sheng particles is stable.
Compared with the prior art, the invention has the beneficial effects that:
(1) The method for establishing the fingerprint of the Fangfengsheng particles is simple, convenient and reliable, high in stability and precision and good in repeatability.
(2) The invention establishes a common mode of the fingerprint spectrum of the Fangfengsheng particles, marks 22 common peaks and identifies 7 common characteristic peaks. The method selects the number 12 peak baicalin as a reference peak in the fingerprint spectrum, and determines the relative retention time of each common peak.
(3) The fingerprint method established in the invention is used for quality monitoring of the Fengsheng particles, and can comprehensively and effectively control the quality of the Fengsheng particles on the whole, thereby ensuring the stability, reliability and safety of the product.
Drawings
Fig. 1 shows the HPLC fingerprint and the control fingerprint (R) of 15 batches of ledebouriella seseloides total holy particles.
Fig. 2 detects wavelength contrast.
FIG. 3 HPLC fingerprint of the mixed control solution and Fangfengsheng particles.
The specific embodiment is as follows:
embodiments of the present invention will be described in detail with reference to the following examples, but the scope of the present invention is not limited to the following examples.
The instruments and reagents used in the examples below were as follows:
1. instrument for measuring and controlling the intensity of light
Agilent 1260 high performance liquid chromatograph; the column was a ZORBAX Eclipse Plus C18 column (5 μm, 4.6X1250 mm). EL204 electronic balance (mertrer-toliter instruments Shanghai limited); SHB-III type circulating water multipurpose vacuum pump (zheng great wall department industry and trade limited); DFY8000 type universal pulverizer (Shanghai bilang instruments, inc.); KQ-500VDB ultrasonic cleaner (Kunshan ultrasonic instruments Co., ltd.).
2. Medicine and reagent
The drugs and reagents used in the following examples: the Fengshutongsheng granule is provided by Shandong Runshen Chinese medicine industry Co., ltd, and has the batch number of: 1803001, 1803002, 1803003, 1803004, 1803005, 1803006, 1803007, 1803008, 1803009, 1803010, 1803011, 1803012, 1803013, 1803014, 1803015. Geniposide (110749-2016617), glycyrrhizin (111610-201607), ammonium glycyrrhizate (110737-201619), baicalin (111595-201607), aloe-emodin (110795-201710), cimicifuga rhizome glycoside (111522-2016712), 5-O-methyl-vitamin amiloride (111523-201610), and the above control are all purchased from Chinese food and drug inspection institute. Acetonitrile is chromatographic purity; methanol and phosphoric acid are analytically pure.
Example 1 fingerprint of Fengshengsheng particles established by high Performance liquid chromatography
1. Preparation of test solution: grinding the Fengshong sheng particles, sieving with a fifth sieve, precisely weighing, placing in a conical flask with a plug, precisely adding 25ml of 70% methanol, weighing, ultrasonically extracting for 30 minutes, cooling, weighing again, supplementing the lost weight with 70% methanol, shaking uniformly, filtering, collecting the subsequent filtrate, and passing through a microporous filter membrane of 0.45 μm.
2. Preparation of a mixed control solution: accurately weighing reference substances including geniposide, cimicifuga rhizome glycoside, glycyrrhizin, 5-O-methylvitamin amiloride, baicalin, ammonium glycyrrhizinate, and aloe-emodin, dissolving in 70% methanol, and respectively preparing into geniposide (50μg.mL) -1 ) Cimicifugal glycoside (20. Mu.g.ml) -1 ) Liquiritigenin (20 mug.ml) -1 ) 5-O-methyl-Weisi-amiloride (10. Mu.g.ml) -1 ) Baicalin (0.4 mg.ml) -1 ) Ammonium glycyrrhizinate (0.20 mg.ml) -1 ) Aloe-emodin (5 mug. Ml) -1 ) Is filtered by a microporous filter membrane with the thickness of 0.45 mu m.
3. And (3) measuring: respectively precisely sucking the sample solution and the mixed reference solution, injecting into a high performance liquid chromatograph, measuring, and recording the chromatogram under the following chromatographic conditions:
chromatographic column: agilent ZORBAX Eclipse Plus C column (5 μm, 4.6X1250 mm); the detection wavelength is 254nm; the flow rate is 1.0ml/min; the column temperature is 25 ℃; the sample injection amount is 10 μl; the mobile phase A is 0.1% phosphoric acid aqueous solution, the mobile phase B is acetonitrile, and a gradient elution mode is adopted, and the gradient elution method is as follows:
4. construction of fingerprint of Fengshong Sheng particles: introducing 15 batches of sample chromatograms of the Fengshen particles into traditional Chinese medicine chromatographic fingerprint similarity evaluation software, selecting common chromatographic peaks in all batches of Fengshen particles, generating a Fengshen particle control fingerprint, taking a No. 12 peak as a reference peak, and calculating the relative peak area and the relative retention time of all the common peaks. Results: relative retention time: peak No. 1 peak 0.1902 ± 0.0003,2 peak 0.3131 ± 0.0014,3 peak 0.4669 ± 0.0019,4 peak 0.5874 ± 0.0020,5 peak 0.6102 ± 0.0020,6 peak 0.6426 ± 0.0019,7 peak 0.6565 ± 0.0017,8 peak 0.6747 ± 0.0016,9 peak 0.7332 ±0.0016, peak 10 0.9459 ±0.0003, peak 11 peak 0.9741 ±0.0003, peak 13 0.9741 ±0.0004, peak 14 peak 0.9741 ±0.0003, peak 15 peak 1.1720±0.0006, peak 16 peak 1.2308±0.0008, peak 17 peak 0.9741 ±0.0010, peak 18 peak 0.9741 ±0.0011, peak 19 peak 0.9741 ±0.0012, peak 20 peak 0.9741 ±0.0014, peak 21 peak 0.9741 ±0.0016, peak 22 peak 0.9741 ±0.0016; relative peak area: peak No. 1, peak No. 0.0581 ± 0.0002,2, peak No. 0.0581 ± 0.0010,3, peak No. 0.0414 ± 0.0007,5, peak No. 0.1306± 0.0003,6, peak No. 0.0373± 0.0005,7, peak No. 0.0232 ± 0.0004,8, peak No. 0.0523±0.0006, peak No. 10, 0427±0.0001, peak No. 11, peak No. 0.0527 ±0.0005, peak No. 13, peak No. 0.0571 ±0.0001, peak No. 14, peak No. 0318+0.0004, peak No. 15, peak No. 0.2252 ±0.0041, peak No. 0.3343 ±0.0006, peak No. 17, peak No. 0.0865 ±0.0002, peak No. 18, peak No. 0.1545 ±0.0002, peak No. 20, peak No. 0.0280±0.0001, peak No. 21, peak No. 0199±0.0002, and peak No. 22, peak No. 0.0248 ±0.0004.
5. The similarity evaluation is carried out on 15 batches of radix sileris Tongsheng particle fingerprints by using a research edition (2012A) of a traditional Chinese medicine chromatographic fingerprint similarity evaluation system, wherein the similarity is more than 0.99, and the results are shown in Table 1.
Example 2 methodological investigation of fingerprint of Fangfengsheng particles
1. Selection of gradient elution mode
The Fengshong sheng granule is a compound preparation consisting of seventeen traditional Chinese medicines, and contains more complex chemical components, so that a gradient elution mode is adopted. Octadecylsilane chemically bonded silica is used as a filler, 0.1% phosphoric acid aqueous solution is used as a mobile phase A, acetonitrile is used as a mobile phase B, the elution flow rate is 1.0ml/min, the column temperature is 25 ℃, the sample injection amount is 10 mu l, and the gradient elution method is as follows:
2. selection of detection wavelength
The number of chromatographic peaks in the chromatograms of the test sample solutions at detection wavelengths of 210nm, 230nm, 254nm, 280nm,300nm,320nm and 350nm are respectively examined, and the results are shown in FIG. 2.
3. Precision test
Taking a sample solution, continuously sampling for 6 times according to the chromatographic conditions, respectively measuring the retention time and the peak area, taking a number 12 peak baicalin peak as a reference peak, calculating the relative retention time and the relative peak area of each common peak and the reference peak, and calculating the RSD value, wherein the results are shown in tables 2 and 3.
TABLE 2 relative retention time of the consensus peaks in the precision test
TABLE 3 relative peak areas of the consensus peaks in the precision test
4. Stability test
The same Fengshong sheng granule test sample solution is taken, measured at 0, 2, 4, 6, 8, 10, 12 and 24 hours according to the chromatographic conditions, the baicalin peak of No. 12 is taken as a reference peak, the relative retention time and the relative peak area of each common peak and the reference peak are calculated, the RSD value is calculated, and the results are shown in tables 4 and 5.
TABLE 4 relative retention time of the individual consensus peaks in the stability test
TABLE 5 relative peak areas of the consensus peaks in stability test
5. Repeatability test
Taking the same batch of Fengshong sheng granule samples, and preparing 6 parts of sample solution according to the preparation method of the sample solution. Measured according to the chromatographic conditions described above. The relative retention time and relative peak area of each common peak and the reference peak were calculated using the number 12 baicalin peak as the reference peak, and RSD values were calculated, and the results are shown in tables 6 and 7.
TABLE 6 relative retention time of individual consensus peaks in the repeatability test
TABLE 7 relative peak areas of the consensus peaks in the stable repeatability test
The above summary and the detailed description are intended to demonstrate practical applications of the technical solutions provided by the present invention, and should not be construed as limiting the scope of the present invention. Various modifications, equivalent alterations, or improvements will occur to those skilled in the art, and are within the spirit and principles of the invention. The scope of the invention is defined by the appended claims.
Claims (3)
1. The method for establishing the fingerprint of the Fangfengsheng granule is characterized by comprising the following steps of:
(1) Preparation of test solution: taking about 2g of Fangfengsheng particles, grinding, sieving with a fifth sieve, precisely weighing, placing into a conical flask with a plug, precisely adding 25ml of 70% methanol, weighing, ultrasonically extracting for 30 minutes, cooling, weighing again, supplementing the lost weight with 70% methanol, shaking uniformly, filtering, collecting the filtrate, and passing through a microporous membrane of 0.45 μm to obtain the final product;
(2) Preparation of a mixed control solution: accurately weighing reference substances of geniposide, cimicifuga rhizome glycoside, glycyrrhizin, 5-O-methylweisi amiloride, baicalin, ammonium glycyrrhizinate and aloe-emodin, adding appropriate amount of solvent, dissolving, and filtering with microporous membrane to obtain reference substance solution;
(3) And (3) measuring: respectively precisely sucking the sample solution and the mixed reference solution, injecting into a high performance liquid chromatograph, measuring, and recording the chromatogram under the following chromatographic conditions:
chromatographic column: an Anjie ZORBAX Eclipse Plus C column, 5 μm, 4.6X1250 mm; the detection wavelength is 254nm; the flow rate is 1.0ml/min; the column temperature is 25 ℃; the sample injection amount is 10 μl; the mobile phase A is 0.1% phosphoric acid aqueous solution, the mobile phase B is acetonitrile, and a gradient elution mode is adopted, and the method is as follows:
(4) Construction of fingerprint of Fengshong Sheng particles: introducing different batches of sample chromatograms into traditional Chinese medicine chromatographic fingerprint similarity evaluation software, selecting common chromatographic peaks in each batch of sample to generate a Fangtong sheng particle control fingerprint, and calculating the relative peak area and relative retention time of each common peak.
2. The method for establishing fingerprint spectrum of Fengshong sheng particles according to claim 1, wherein in the step (2), the preparation method of the mixed reference substance solution is as follows: respectively weighing geniposide, cimicifuga rhizome glycoside, glycyrrhizin, 5-O-methyl vitamin amiloride, baicalin, ammonium glycyrrhizinate, and aloe-emodin reference substances, adding 70% methanol, dissolving, and making into 50 μg ml -1 Gardenia jasminoides ellis of (20 μg ml) -1 Cimicifugal glycoside of (A), 20 μg.ml -1 Glycyrrhizin of (C), 10 μg.ml -1 5-O-methylvisamiloride, 0.4 mg.ml) -1 Baicalin of (0.20 mg.ml) -1 Ammonium glycyrrhetate of (5. Mu.g.ml) -1 The aloe-emodin mixed reference substance solution is filtered by a microporous filter membrane with the thickness of 0.45 mu m.
3. The method for establishing the fingerprint of the Fengshengsheng particles according to claim 1, wherein in the step (4), the fingerprint obtained by the establishment method contains 22 characteristic peaks, wherein No. 2 peak, no. 3 peak, no. 5 peak, no. 9 peak, no. 12 peak, no. 19 peak and No. 20 peak are respectively gardenin, cimicifugal glucoside, glycyrrhizin, 5-O-methylweis amiloride, baicalin, ammonium glycyrrhizate and aloe-emodin, and the relative retention time and relative peak area of the other 21 characteristic peaks are calculated by taking the baicalin of the No. 12 peak as a reference peak: relative retention time: peak No. 1 peak 0.1902 ± 0.0003,2 peak 0.3131 ± 0.0014,3 peak 0.4669 ± 0.0019,4 peak 0.5874 ± 0.0020,5 peak 0.6102 ± 0.0020,6 peak 0.6426 ± 0.0019,7 peak 0.6565 ± 0.0017,8 peak 0.6747 ± 0.0016,9 peak 0.7332 ±0.0016, peak 10 0.9459 ±0.0003, peak 11 peak 0.9741 ±0.0003, peak 13 0.9741 ±0.0004, peak 14 peak 0.9741 ±0.0003, peak 15 peak 1.1720±0.0006, peak 16 peak 1.2308±0.0008, peak 17 peak 0.9741 ±0.0010, peak 18 peak 0.9741 ±0.0011, peak 19 peak 0.9741 ±0.0012, peak 20 peak 0.9741 ±0.0014, peak 21 peak 0.9741 ±0.0016, peak 22 peak 0.9741 ±0.0016; relative peak area: peak No. 1, peak No. 0.0581 ± 0.0002,2, peak No. 0.0581 ± 0.0010,3, peak No. 0.0414 ± 0.0007,5, peak No. 0.1306± 0.0003,6, peak No. 0.0373± 0.0005,7, peak No. 0.0232 ± 0.0004,8, peak No. 0.0523±0.0006, peak No. 10, 0427±0.0001, peak No. 11, peak No. 0.0527 ±0.0005, peak No. 13, peak No. 0.0571 ±0.0001, peak No. 14, peak No. 0318+0.0004, peak No. 15, peak No. 0.2252 ±0.0041, peak No. 0.3343 ±0.0006, peak No. 17, peak No. 0.0865 ±0.0002, peak No. 18, peak No. 0.1545 ±0.0002, peak No. 20, peak No. 0.0280±0.0001, peak No. 21, peak No. 0199±0.0002, and peak No. 22, peak No. 0.0248 ±0.0004.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1895408A (en) * | 2006-06-27 | 2007-01-17 | 天津中新药业集团股份有限公司达仁堂制药厂 | Radix saposhnikoviae Tong Sheng capsules, preparation and quality control thereof |
| CN1957999A (en) * | 2005-10-31 | 2007-05-09 | 成都华神集团股份有限公司 | Composition of Chinese traditional medicine, preparation method, and quality control method |
-
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1957999A (en) * | 2005-10-31 | 2007-05-09 | 成都华神集团股份有限公司 | Composition of Chinese traditional medicine, preparation method, and quality control method |
| CN1895408A (en) * | 2006-06-27 | 2007-01-17 | 天津中新药业集团股份有限公司达仁堂制药厂 | Radix saposhnikoviae Tong Sheng capsules, preparation and quality control thereof |
Non-Patent Citations (7)
| Title |
|---|
| HPLC法测定防风通圣颗粒中盐酸麻黄碱和盐酸伪麻黄碱含量;邱燕;;海峡药学(第01期);全文 * |
| RP-HPLC法测定防风通圣丸中栀子苷、芍药苷、黄芩苷、大黄素的含量;杨小军;方博文;丁永辉;;西部中医药(第09期);全文 * |
| RP-HPLC法测定防风通圣丸中芍药苷、栀子苷、黄芩苷和连翘苷的含量;刘阿静;齐广才;刘珍叶;白梓静;;西北药学杂志(第05期);全文 * |
| SPE-UPLC法同时测定复方荆防颗粒中的升麻素苷和5-O-甲基维斯阿米醇苷的含量;江丽;余辉;曹月霞;龚春燕;;中国医药导报(第01期);全文 * |
| 双波长HPLC同时测定防风通圣丸中栀子苷和黄芩苷的含量;侯志坚;师永清;;中国实验方剂学杂志(第24期);全文 * |
| 防风通圣丸(颗粒)质量研究;史宪海;郭景文;李民生;于荣;李萌;;解放军药学学报(第06期);全文 * |
| 防风通圣丸质量标准研究;尹浣姝;宋新波;刘晓明;张丽娟;;现代中药研究与实践(第01期);全文 * |
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