CN113237961A - Method for detecting main drug effect components of cinnamon and storax in pharmaceutical preparation - Google Patents
Method for detecting main drug effect components of cinnamon and storax in pharmaceutical preparation Download PDFInfo
- Publication number
- CN113237961A CN113237961A CN202110043810.5A CN202110043810A CN113237961A CN 113237961 A CN113237961 A CN 113237961A CN 202110043810 A CN202110043810 A CN 202110043810A CN 113237961 A CN113237961 A CN 113237961A
- Authority
- CN
- China
- Prior art keywords
- storax
- cinnamon
- pharmaceutical preparation
- phase
- detecting
- 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.)
- Pending
Links
- 241001060310 Styracaceae Species 0.000 title claims abstract description 47
- 235000001361 Styrax officinalis Nutrition 0.000 title claims abstract description 47
- 235000019382 gum benzoic Nutrition 0.000 title claims abstract description 47
- 239000000825 pharmaceutical preparation Substances 0.000 title claims abstract description 46
- 244000223760 Cinnamomum zeylanicum Species 0.000 title claims abstract description 45
- 235000017803 cinnamon Nutrition 0.000 title claims abstract description 45
- 238000000034 method Methods 0.000 title claims abstract description 31
- 230000000857 drug effect Effects 0.000 title description 5
- WBYWAXJHAXSJNI-UHFFFAOYSA-N methyl p-hydroxycinnamate Natural products OC(=O)C=CC1=CC=CC=C1 WBYWAXJHAXSJNI-UHFFFAOYSA-N 0.000 claims abstract description 65
- 235000013985 cinnamic acid Nutrition 0.000 claims abstract description 64
- WBYWAXJHAXSJNI-VOTSOKGWSA-M .beta-Phenylacrylic acid Natural products [O-]C(=O)\C=C\C1=CC=CC=C1 WBYWAXJHAXSJNI-VOTSOKGWSA-M 0.000 claims abstract description 63
- 229930016911 cinnamic acid Natural products 0.000 claims abstract description 63
- WBYWAXJHAXSJNI-SREVYHEPSA-N Cinnamic acid Chemical compound OC(=O)\C=C/C1=CC=CC=C1 WBYWAXJHAXSJNI-SREVYHEPSA-N 0.000 claims abstract description 62
- 238000004128 high performance liquid chromatography Methods 0.000 claims abstract description 32
- 238000002360 preparation method Methods 0.000 claims abstract description 28
- 239000003960 organic solvent Substances 0.000 claims abstract description 21
- 239000012085 test solution Substances 0.000 claims abstract description 19
- 238000002137 ultrasound extraction Methods 0.000 claims abstract description 14
- 239000000706 filtrate Substances 0.000 claims abstract description 9
- 238000001914 filtration Methods 0.000 claims abstract description 7
- 238000001816 cooling Methods 0.000 claims abstract description 4
- 230000003285 pharmacodynamic effect Effects 0.000 claims abstract 2
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 57
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 33
- 238000001514 detection method Methods 0.000 claims description 33
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 30
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 27
- 239000000243 solution Substances 0.000 claims description 22
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 15
- 239000013558 reference substance Substances 0.000 claims description 14
- 239000012088 reference solution Substances 0.000 claims description 12
- 238000010828 elution Methods 0.000 claims description 9
- 239000006187 pill Substances 0.000 claims description 9
- 238000004458 analytical method Methods 0.000 claims description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 7
- 238000010812 external standard method Methods 0.000 claims description 5
- 238000002347 injection Methods 0.000 claims description 5
- 239000007924 injection Substances 0.000 claims description 5
- 230000014759 maintenance of location Effects 0.000 claims description 5
- 239000000203 mixture Substances 0.000 claims description 4
- 239000002904 solvent Substances 0.000 claims description 4
- 239000004480 active ingredient Substances 0.000 claims description 2
- 239000004615 ingredient Substances 0.000 claims 1
- 238000003908 quality control method Methods 0.000 abstract description 6
- 239000012488 sample solution Substances 0.000 description 19
- 239000000523 sample Substances 0.000 description 17
- 239000003814 drug Substances 0.000 description 12
- 239000007864 aqueous solution Substances 0.000 description 9
- 230000000694 effects Effects 0.000 description 7
- 238000012360 testing method Methods 0.000 description 6
- 239000007788 liquid Substances 0.000 description 5
- 239000000843 powder Substances 0.000 description 5
- 229940079593 drug Drugs 0.000 description 4
- 238000005259 measurement Methods 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 238000004364 calculation method Methods 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 239000012528 membrane Substances 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 208000024891 symptom Diseases 0.000 description 3
- 238000005303 weighing Methods 0.000 description 3
- 206010002383 Angina Pectoris Diseases 0.000 description 2
- 125000003118 aryl group Chemical group 0.000 description 2
- 238000000227 grinding Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 208000031225 myocardial ischemia Diseases 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 229910021642 ultra pure water Inorganic materials 0.000 description 2
- 239000012498 ultrapure water Substances 0.000 description 2
- 239000003643 water by type Substances 0.000 description 2
- 230000004580 weight loss Effects 0.000 description 2
- DTGKSKDOIYIVQL-WEDXCCLWSA-N (+)-borneol Chemical compound C1C[C@@]2(C)[C@@H](O)C[C@@H]1C2(C)C DTGKSKDOIYIVQL-WEDXCCLWSA-N 0.000 description 1
- REPVLJRCJUVQFA-UHFFFAOYSA-N (-)-isopinocampheol Natural products C1C(O)C(C)C2C(C)(C)C1C2 REPVLJRCJUVQFA-UHFFFAOYSA-N 0.000 description 1
- JHGWQSGWUPCKNT-UHFFFAOYSA-N 2-tert-butyl-4-methyl-1,3,5-trinitrobenzene Chemical compound CC1=C([N+]([O-])=O)C=C([N+]([O-])=O)C(C(C)(C)C)=C1[N+]([O-])=O JHGWQSGWUPCKNT-UHFFFAOYSA-N 0.000 description 1
- 206010002660 Anoxia Diseases 0.000 description 1
- 241000976983 Anoxia Species 0.000 description 1
- 201000001320 Atherosclerosis Diseases 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 206010004542 Bezoar Diseases 0.000 description 1
- 206010008479 Chest Pain Diseases 0.000 description 1
- 244000037364 Cinnamomum aromaticum Species 0.000 description 1
- 235000014489 Cinnamomum aromaticum Nutrition 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 241000402754 Erythranthe moschata Species 0.000 description 1
- 206010021143 Hypoxia Diseases 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 239000003088 amphibian venom Substances 0.000 description 1
- 230000007953 anoxia Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 229940116229 borneol Drugs 0.000 description 1
- CKDOCTFBFTVPSN-UHFFFAOYSA-N borneol Natural products C1CC2(C)C(C)CC1C2(C)C CKDOCTFBFTVPSN-UHFFFAOYSA-N 0.000 description 1
- 230000007211 cardiovascular event Effects 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 208000029078 coronary artery disease Diseases 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- DTGKSKDOIYIVQL-UHFFFAOYSA-N dl-isoborneol Natural products C1CC2(C)C(O)CC1C2(C)C DTGKSKDOIYIVQL-UHFFFAOYSA-N 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000020710 ginseng extract Nutrition 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000007721 medicinal effect Effects 0.000 description 1
- 208000010125 myocardial infarction Diseases 0.000 description 1
- 150000002823 nitrates Chemical class 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- 208000023482 precordial pain Diseases 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 238000004506 ultrasonic cleaning Methods 0.000 description 1
- 230000029663 wound healing Effects 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
- G01N30/62—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
-
- 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
- G01N2030/042—Standards
- G01N2030/047—Standards external
Landscapes
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Pathology (AREA)
- Biochemistry (AREA)
- Immunology (AREA)
- General Physics & Mathematics (AREA)
- Library & Information Science (AREA)
- Engineering & Computer Science (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
The invention provides a method for detecting main pharmacodynamic components of cinnamon and storax in a pharmaceutical preparation, which comprises the following steps: dissolving a medicinal preparation sample in an organic solvent, performing ultrasonic extraction, cooling, shaking up, filtering, taking a test solution obtained from a subsequent filtrate, and detecting by High Performance Liquid Chromatography (HPLC) to determine the common main effective components of cinnamon and storax in the test solution: the content of cinnamic acid. The method for detecting the main medicinal components of the cinnamon and the storax in the medicinal preparation has the advantages of good precision, reproducibility and stability, accuracy and reliability, can truly reflect the quality difference of the cinnamon and the storax in the medicinal preparation, and comprehensively improves the quality control system of the medicinal preparation.
Description
Technical Field
The invention belongs to the technical field of detection of traditional Chinese medicine components, relates to a detection method of main drug effect components of cinnamon and storax in a pharmaceutical preparation, and particularly relates to a method for detecting the common main drug effect components of the cinnamon and the storax in the pharmaceutical preparation, which comprises the following steps: a method for detecting cinnamic acid.
Background
The medicinal preparation is improved from Suhe flavor pills in Song Dynasty in the 70 s, is one of the most commonly used treatment medicaments for the coronary heart disease at present by using the advantages of definite curative effect, safety and economy, has quick symptom relieving and effect taking, does not have contraindication and drug resistance of nitrates, can protect blood vessels after being taken for a long time, prevent atherosclerosis, reduce the attack frequency of angina and reduce the occurrence of serious cardiovascular events, has aromatic and warm effects, tonifies qi and strengthens heart, is clinically used for chest stuffiness caused by qi stagnation and blood stasis, and has the symptoms of precordial pain and immobility; angina pectoris and myocardial infarction caused by myocardial ischemia are the above symptoms. Because the curative effect is excellent and the safety is high, the Chinese medicine is classified as a national Chinese medicine secret variety and protected by the national secret technology, and the listed national basic medicine catalogue honors a plurality of honors such as the scientific and technical prize of the Chinese traditional and western medicine combination society.
The medicinal preparation is black brown and glossy pellet, has tan cross section after being crushed, and is bitter, pungent and cool in taste and numb and tongue feeling. The medicine preparation consists of 7 kinds of Chinese medicinal materials, including artificial musk, ginseng extract, storax, artificial bezoar, cassia, toad venom, borneol, etc. and contains both non-volatile component and volatile component. The cinnamon and the storax are used as traditional Chinese medicinal materials with the effects of aromatic dredging collaterals, the common main medicinal component is cinnamic acid, and the cinnamic acid has the effects of resisting inflammation, resisting bacteria, promoting wound healing, improving myocardial ischemia and anoxia damage and the like. Therefore, the method selects the cinnamic acid as an index component for measuring the content of the cinnamon and the storax in the pharmaceutical preparation. Therefore, it is necessary to detect the quality control of cinnamon and storax and perfect the quality control system of the pharmaceutical preparation.
Disclosure of Invention
In view of the above-mentioned drawbacks of the prior art, the present invention aims to provide a method for detecting the main effective components of cinnamon and storax in a pharmaceutical preparation, which is used to solve the problem that the prior art lacks the main effective components common to the cinnamon and storax components of the pharmaceutical preparation: the problem of the content determination method of the cinnamic acid.
In order to achieve the above and other related objects, a first aspect of the present invention provides a method for detecting main effective components of cinnamon and storax in a pharmaceutical preparation, comprising: adding an organic solvent into a medicinal preparation sample for dissolving, performing ultrasonic extraction, cooling, shaking up, filtering, taking a test sample solution obtained from a subsequent filtrate, and detecting by adopting High Performance Liquid Chromatography (HPLC) to determine main common effective components of cinnamon and storax in the test sample solution: the content of cinnamic acid.
Preferably, the CAS number of the cinnamic acid is 140-10-3.
Preferably, the pharmaceutical preparation sample is a ground pharmaceutical preparation powder.
Preferably, the ratio of the added mass of the drug preparation sample to the added volume of the organic solvent is 1:20-50 g/mL. Preferably, the ratio of the mass of the drug preparation sample added to the volume of the organic solvent added is 1:20, g/mL.
Preferably, the organic solvent is selected from one of methanol, ethanol, acetonitrile, 95% ethanol, and 80% methanol. Preferably, the organic solvent is 80% methanol. The 95% ethanol is 95% ethanol water solution by volume percentage. The 80% methanol is 80% methanol aqueous solution by volume percentage.
Preferably, the ultrasonic extraction time is 10-40 min. Preferably, the ultrasound extraction time is 20 min.
Preferably, the power of the ultrasonic extraction is 250-350W, and the frequency of the ultrasonic extraction is 35-45 kHz. Preferably, the power of the ultrasonic extraction is 300W, and the frequency of the ultrasonic extraction is 40 kHz.
Preferably, the pharmaceutical preparation sample is precisely weighed after the addition of the organic solvent.
Preferably, the cooling is followed by reweighing and top-up loss with organic solvent.
Preferably, the filtration means: filtering the supernatant of the shaken solution with a filter membrane, and discarding the primary filtrate to obtain the subsequent filtrate.
More preferably, the filter is a 0.45 μm filter.
Preferably, the detection is performed by High Performance Liquid Chromatography (HPLC), comprising the steps of:
1) preparing a reference solution: adding an organic solvent into a cinnamic acid reference substance for dissolving and fixing the volume to prepare a reference substance solution;
2) sample detection: respectively detecting the test solution and the reference solution in the step 1) by adopting a High Performance Liquid Chromatography (HPLC), comparing the retention time for qualitative determination, and determining the content of the cinnamic acid in the test solution by adopting an external standard method for quantitative determination.
Preferably, in step 1), the organic solvent is selected from one of methanol, ethanol, acetonitrile, 95% ethanol, and 80% methanol. . More preferably, the organic solvent is 80% methanol.
Preferably, in the step 1), the content of the cinnamic acid in the control solution is in the range of 5.34-222.40 μ g/ml.
Preferably, in step 1), the reference solution is prepared by stepwise dilution.
Preferably, in the step 2), the detector used in the High Performance Liquid Chromatography (HPLC) is a photodiode array detector (DAD).
Preferably, in the step 2), the chromatographic column in the high performance liquid chromatography is a T3 chromatographic column. More preferably, the column in the high performance liquid chromatography is a Waters X-select HSS T3 column (column length 250mm, inner diameter 4.6mm, particle size 5 μm).
Preferably, in the step 2), the detection wavelength in the high performance liquid chromatography is 285-295 nm. More preferably, the detection wavelength in the high performance liquid chromatography is 290 nm.
Preferably, in step 2), the column temperature in the high performance liquid chromatography is 25-35 ℃. More preferably, the column temperature in the high performance liquid chromatography is 25 ℃.
Preferably, in the step 2), the flow rate in the high performance liquid chromatography is 0.8-1.2 ml/min. More preferably, the flow rate in the high performance liquid chromatography is 0.8 ml/min.
Preferably, in the step 2), the sample injection amount in the high performance liquid chromatography is 5-15 μ l. More preferably, the sample amount in the high performance liquid chromatography is 10. mu.l.
Preferably, in the step 2), in the high performance liquid chromatography, the mobile phase is acetonitrile-0.02-0.04% phosphoric acid aqueous solution, wherein the phase A is acetonitrile, and the phase B is 0.02-0.04% phosphoric acid aqueous solution; the analysis time is 50 min; gradient elution.
More preferably, in the high performance liquid chromatography, the mobile phase is acetonitrile-0.03% phosphoric acid aqueous solution, wherein, the phase A is acetonitrile, and the phase B is 0.03% phosphoric acid aqueous solution; the analysis time is 50 min; gradient elution.
The 0.02-0.04% phosphoric acid aqueous solution is 0.02-0.04% phosphoric acid aqueous solution by volume percentage. The 0.03% phosphoric acid aqueous solution is 0.03% phosphoric acid aqueous solution by volume percentage.
More preferably, as shown in table 1, the specific procedure of the gradient elution is:
0-25 min, phase A: the volume ratio of the phase B is 20: 80-40: 60, adding a solvent to the mixture;
25-35 min, phase A: the volume ratio of the phase B is 40: 60-50: 50;
35-40 min, phase A: the volume ratio of the phase B is 50: 50-100: 0;
40-50 min, phase A: the volume ratio of the phase B is 100: 0-100: 0.
TABLE 1
Preferably, in step 2), the external standard method comprises the following steps:
A) preparing a series of reference substance solutions with different concentrations according to the step 1), respectively carrying out HPLC detection to obtain a linear relation between the chromatographic peak area of cinnamic acid and the content of corresponding cinnamic acid, drawing a corresponding standard working curve, and calculating to obtain a regression equation of the standard working curve of the cinnamic acid;
B) and (3) carrying out HPLC detection on the test solution, substituting the obtained chromatographic peak area of the cinnamic acid into the regression equation of the corresponding standard working curve of the cinnamic acid in the step A), and calculating to obtain the content of the cinnamic acid in the test solution.
More preferably, the standard working curve has a chromatographic peak area of cinnamic acid as ordinate (Y-axis) and its corresponding content of cinnamic acid as abscissa (X-axis).
The second aspect of the invention provides an application of a detection method of main effective components of cinnamon and storax in a pharmaceutical preparation in quality detection of the cinnamon and the storax in the pharmaceutical preparation.
The medicinal preparation is a medicinal preparation containing cinnamon and storax, and specifically includes but is not limited to musk heart-protecting pills and the like.
The third aspect of the invention provides a quality detection method for cinnamon and storax in a pharmaceutical preparation, wherein the cinnamon and storax contained in the pharmaceutical preparation are more than or equal to 0.092% in terms of cinnamic acid.
Preferably, the weight of each pill of the pharmaceutical preparation is 22.5mg, and the content of cinnamon and storax in each pill of the pharmaceutical preparation is more than or equal to 21 mu g calculated by cinnamic acid.
As described above, the method for detecting the main effective components of cinnamon and storax in the pharmaceutical preparation provided by the present invention adopts the pretreatment and instrumental detection methods of optimized conditions to detect the main effective components of cinnamon and storax in the pharmaceutical preparation: and (4) carrying out accurate quantitative and qualitative detection on the cinnamic acid. The method has good precision, reproducibility and stability, is accurate and reliable, can truly reflect the quality difference of the cinnamon and the storax in the pharmaceutical preparation, ensures the stability of the production process and quality among batches, is used for supplementing and improving the current quality control standard, and comprehensively perfects the quality control system of the pharmaceutical preparation.
Drawings
Fig. 1 shows the main active ingredients shared by cinnamon and storax components in the present invention: high performance liquid chromatograms 1a and 1b of a reference substance and a test substance of cinnamic acid, wherein fig. 1a is the high performance liquid chromatogram of the reference substance, and fig. 1b is the high performance liquid chromatogram of the test substance; reference character a is cinnamic acid.
Detailed Description
The present invention is further illustrated below with reference to specific examples, which are intended to be illustrative only and not to limit the scope of the invention.
The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification. The invention is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope of the present invention.
The reagents and equipment used in the following examples are as follows:
1. reagent
Pharmaceutical preparations (Shanghai and Huang pharmaceuticals, Inc.); cinnamic acid standard (China institute for testing and testing food and drug, purity 98% or more; batch number 110786-; acetonitrile (chromatographic grade, TEDIA); phosphoric acid (chromatographic grade, TEDIA); the rest of the reagents (analytically pure, Shanghai Zhanyun chemical Co., Ltd.); deionized water (Mili-Q ultrapure water preparation).
2. Instrument for measuring the position of a moving object
1260 high performance liquid chromatograph, configured with on-line vacuum degasser, autosampler, quaternary pump, DAD detector, thermostatted column oven; agilent ChemStation C01.05 chromatography workstation (Agilent Corp.); SB-5200 ultrasonic cleaning apparatus (Ningbo Xinzhi Biotech Co., Ltd.); Mili-Q ultra pure water instruments (Milipore Corp.).
The content of the main effective components of cinnamon and storax in the pharmaceutical preparation comprises the following determination process.
1. Preparation of test solution
Grinding the medicinal preparation sample into powder, adding organic solvent for dissolving, and precisely weighing. Wherein the ratio of the added mass of the drug preparation sample to the added volume of the organic solvent is 1:20-50 g/mL. Then, ultrasonic extraction is performed for 10-40 min at the power of 250-350W and the frequency of 35-45 kHz. Cooled, reweighed, and made up to weight loss with organic solvent. Shaking, collecting supernatant, filtering with 0.45 μm filter membrane, and discarding the primary filtrate to obtain subsequent filtrate as sample solution. The organic solvent is selected from one of methanol, ethanol, acetonitrile, 95% ethanol and 80% methanol.
2. Preparation of control solutions
Adding an organic solvent into a cinnamic acid reference substance for dissolving and fixing the volume to prepare a reference substance solution. The content of cinnamic acid in the control solution is 5.34-222.40 μ g/ml.
The organic solvent is selected from one of methanol, ethanol, acetonitrile, 95% ethanol and 80% methanol.
3. Measurement of
Detecting the sample solution and a series of reference solutions with different concentrations by High Performance Liquid Chromatography (HPLC), comparing the retention time, and quantifying by external standard method. A series of reference substance solutions with different concentrations are respectively subjected to HPLC detection to obtain a linear relation between the chromatographic peak area of the cinnamic acid and the content of the corresponding cinnamic acid, a corresponding standard working curve is drawn, and a regression equation of the standard working curve of the cinnamic acid is obtained through calculation. And carrying out HPLC detection on the test solution, substituting the obtained chromatographic peak area of the cinnamic acid into a regression equation of a corresponding standard working curve of the cinnamic acid, and calculating to obtain the content of the cinnamic acid in the test solution.
Wherein, the high performance liquid chromatography comprises the following detection conditions:
the detector is a photodiode array detector (DAD); the chromatographic column is a T3 chromatographic column; the detection wavelength is 285-295 nm; the column temperature is 25-35 ℃; the flow rate is 0.8-1.2 ml/min; the sample injection amount is 5-15 mu l; the mobile phase is acetonitrile-0.02-0.04% phosphoric acid water solution, wherein, the phase A is acetonitrile, and the phase B is 0.02-0.04% phosphoric acid water solution; the analysis time is 50 min; gradient elution.
The specific procedure for gradient elution was:
0-25 min, phase A: the volume ratio of the phase B is 20: 80-40: 60, adding a solvent to the mixture;
25-35 min, phase A: the volume ratio of the phase B is 40: 60-50: 50;
35-40 min, phase A: the volume ratio of the phase B is 50: 50-100: 0;
40-50 min, phase A: the volume ratio of the phase B is 100: 0-100: 0.
example 1
1. Preparation of test solution
Grinding 120 pills of the medicinal preparation sample into powder, precisely weighing 1.0g of the powder, placing the powder into a conical flask with a plug, precisely adding 20ml of 80% methanol for dissolution, sealing the plug and precisely weighing. Then ultrasonic extraction is carried out for 20min under the power of 300W and the frequency of 40 kHz. Cooled, reweighed and made up to weight loss with 80% methanol. Shaking, collecting supernatant, filtering with 0.45 μm filter membrane, discarding the initial filtrate to obtain continuous filtrate, which is sample solution No. 1.
2. Preparation of control solutions
Adding methanol into cinnamic acid reference substance, dissolving, and diluting to desired volume to obtain reference solution No. 1. The content of cinnamic acid in the control solution 1# is 5.34-222.40 μ g/ml.
3. Measurement of
Detecting the sample solution 1# and a series of reference solution 1# with different concentrations by High Performance Liquid Chromatography (HPLC), comparing retention time for qualitative determination, and quantifying by external standard method. A series of reference substance solutions 1# with different concentrations are respectively subjected to HPLC detection to obtain a linear relation between the chromatographic peak area of the cinnamic acid and the content of the corresponding cinnamic acid, a corresponding standard working curve is drawn, and a regression equation of the standard working curve of the cinnamic acid is obtained through calculation. And performing HPLC detection on the sample solution 1#, substituting the obtained chromatographic peak area of the cinnamic acid into a regression equation of a corresponding standard working curve of the cinnamic acid, and calculating to obtain the content of the cinnamic acid in the sample solution 1 #.
Wherein, the high performance liquid chromatography comprises the following detection conditions:
the detector is a photodiode array detector (DAD); the column was a Waters X-select HSS T3 column (column length 250mm, inner diameter 4.6mm, particle size 5 μm); the detection wavelength is 290 nm; the column temperature was 25 ℃; the flow rate is 0.8 ml/min; the sample amount is 10 mul; the mobile phase is acetonitrile-0.03% phosphoric acid water solution, wherein the phase A is acetonitrile, and the phase B is 0.03% phosphoric acid water solution; the analysis time is 50 min; gradient elution.
The specific procedure for gradient elution was:
0-25 min, phase A: the volume ratio of the phase B is 20: 80-40: 60, adding a solvent to the mixture;
25-35 min, phase A: the volume ratio of the phase B is 40: 60-50: 50;
35-40 min, phase A: the volume ratio of the phase B is 50: 50-100: 0;
40-50 min, phase A: the volume ratio of the phase B is 100: 0-100: 0.
example 2
A sample of the pharmaceutical formulation of batch No. H160350 is taken and the test solution is prepared in step 1 of example 1. Meanwhile, a reference solution was prepared in step 2 of example 1, wherein the cinnamic acid content in the reference solution was 60 μ g/ml. Respectively and precisely sucking 10 mu l of reference solution and test solution, injecting into a liquid chromatograph, measuring according to the step 2 in the example 1, comparing the retention time, and performing qualitative determination, specifically referring to the figure 1, determining the peak appearance condition of the cinnamic acid in the test solution, wherein the specific data is shown in the table 2.
TABLE 2
Example 3
The common main drug effect components of the cinnamon and the storax in the pharmaceutical preparation of the invention are as follows: the detection method of the cinnamic acid is verified by methodology, and the performance index result is as follows.
1. Precision degree
Taking the pharmaceutical preparation (batch number: 180501), preparing a sample solution according to the step 1 in the example 1, precisely sucking 10 mu l of the same sample solution, injecting the sample solution into a gas chromatograph, continuously feeding and analyzing for 6 times according to the chromatographic conditions of the step 3 in the example 1, measuring the peak area, and calculating the content of cinnamic acid, wherein the specific precision result is shown in the table 3. The results show that the chromatographic peak areas RSD of the cinnamic acid are all less than 0.5%, which indicates that the method has good precision.
TABLE 3
2. Repeatability of
6 parts of the same batch of the pharmaceutical preparation (batch No. 180501) are taken, a sample solution is prepared according to the step 1 in the example 1, 10 mu l of the sample solution is precisely absorbed and injected into a gas chromatograph, sample injection analysis is respectively carried out according to the chromatographic conditions of the step 3 in the example 1, the peak area is measured, and the specific repeatability result is shown in Table 4. The results show that the chromatographic peak areas RSD of the cinnamic acid are all less than 1%, which indicates that the method has good repeatability.
TABLE 4
3. Stability of
Taking the pharmaceutical preparation (batch number: 180501), preparing a sample solution according to the step 1 in the example 1, precisely sucking 10 mu l of the same sample solution, injecting the sample solution into a gas chromatograph, injecting the sample solution into the gas chromatograph for analysis respectively for 0, 4, 8, 12, 24 and 48 hours according to the chromatographic conditions of the step 3 in the example 1, measuring the peak area, calculating the content of cinnamic acid, and the specific stability result is shown in the table 5. The result shows that the chromatographic peak area RSD of the cinnamic acid is less than 2%, and the sample solution is basically stable within 0-48 hours, which indicates that the method has good stability.
TABLE 5
4. Linear relationship of detection method
An appropriate amount of cinnamic acid reference substances are precisely weighed, and methanol is added according to the step 2 in the example 1 to prepare a series of reference substance solutions with different concentrations. According to the chromatographic conditions of the step 3 in the example 1, 10 μ l of the reference solution is precisely sucked and injected into a gas chromatograph, the concentration (μ g/ml) of the reference is taken as the abscissa and the peak area is taken as the ordinate to perform plotting, and the standard regression equation, the correlation coefficient and the linear range of the cinnamic acid are obtained through measurement and calculation, and the specific results are shown in the table 6. As can be seen from Table 6, cinnamic acid showed good linear relationship (r) in the concentration range examined2＝1.0000)。
TABLE 6
5. Recovery rate
6 parts of a pharmaceutical preparation (lot: 180501) of known concentration (0.5 g) were weighed out precisely, and a predetermined amount of 20ml of an 80% methanol solution of a cinnamic acid control was added accurately, and a sample solution was prepared according to step 1 of example 1, and subjected to sample injection analysis according to the chromatographic conditions of step 3 of example 1, and the results are shown in Table 7. As can be seen from Table 7, the average sample recovery rate of cinnamic acid was 102.61% and the RSD value was 0.83%, respectively, and the recovery rates of the measurement results were good, indicating that the method is more accurate.
TABLE 7
6. Limit of detection (LOD) and limit of quantitation (LOQ)
The limit of detection (LOD) is the concentration of the substance to be measured at a signal-to-noise ratio (S/N) of 3; the limit of quantitation (LOQ) is the concentration of the measured substance at a signal-to-noise ratio (S/N) of 10. The results are shown in Table 8, and the LOD of the cinnamic acids is 3.3 mug/ml respectively; LOQ was 2.8. mu.g/ml.
TABLE 8
Example 3
15 batches (180309-171217) of pharmaceutical preparation samples produced in 2018 are collected, a test solution is prepared according to the step 1 in the example 1, and the detection is carried out according to the chromatographic conditions in the step 3 in the example 1. As a result, as shown in Table 9, the content of cinnamic acid was 0.118 to 0.141%. The minimum 70% of the total content of cinnamon and storax in the pharmaceutical preparation is determined by that the pharmaceutical preparation contains cinnamon and storax not less than 0.092% of cinnamic acid. The weight of each pill of the medicinal preparation is converted into 22.5mg, namely the content of cinnamon and storax in each pill of the medicinal preparation is not less than 21 mu g based on the content of cinnamic acid.
TABLE 9
In conclusion, the detection method for the main medicinal effect components of the cinnamon and the storax in the medicinal preparation provided by the invention has the advantages of good precision density, reproducibility and stability, accuracy and reliability, can truly reflect the quality difference of the cinnamon and the storax in the medicinal preparation, and comprehensively improves the quality control system of the medicinal preparation. Therefore, the invention effectively overcomes various defects in the prior art and has high industrial utilization value.
The foregoing embodiments are merely illustrative of the principles and utilities of the present invention and are not intended to limit the invention. Any person skilled in the art can modify or change the above-mentioned embodiments without departing from the spirit and scope of the present invention. Accordingly, it is intended that all equivalent modifications or changes which may be accomplished by those skilled in the art without departing from the spirit and scope of the present invention as set forth in the appended claims.
Claims (10)
1. A method for detecting main effective components of cortex Cinnamomi and storax in pharmaceutical preparation comprises: dissolving a medicinal preparation sample in an organic solvent, performing ultrasonic extraction, cooling, shaking up, filtering, taking a test solution obtained from a subsequent filtrate, and detecting by adopting a high performance liquid chromatography to determine the common main medicinal components of the cinnamon and the storax in the test solution: the content of cinnamic acid.
2. The method for detecting the main effective components of cinnamon and storax in a pharmaceutical preparation according to claim 1, wherein the preparation of the test solution comprises any one or more of the following conditions:
A) the ratio of the added mass of the medicinal preparation sample to the added volume of the organic solvent is 1:20-50, g/mL;
B) the organic solvent is selected from one of methanol, ethanol, acetonitrile, 95% ethanol and 80% methanol;
C) the ultrasonic extraction time is 10-40 min;
D) the power of the ultrasonic extraction is 250-350W, and the frequency of the ultrasonic extraction is 35-45 kHz.
3. The method for detecting the main effective components of cinnamon and storax in the pharmaceutical preparation according to claim 1, wherein the detection is performed by high performance liquid chromatography, comprising the following steps:
1) preparing a reference solution: adding an organic solvent into a cinnamic acid reference substance for dissolving and fixing the volume to prepare a reference substance solution;
2) sample detection: respectively detecting the test solution and the reference solution in the step 1) by adopting a high performance liquid chromatography, comparing the retention time for qualitative determination, and determining the content of cinnamic acid in the test solution by adopting an external standard method for quantitative determination.
4. The method for detecting the main effective components of cinnamon and storax in a pharmaceutical preparation according to claim 3, wherein in step 1), the content of cinnamic acid in the control solution is in the range of 5.34-222.40 μ g/ml.
5. The method for detecting the major pharmacodynamic ingredients of cinnamon and storax in a pharmaceutical preparation according to claim 3, wherein the detection wavelength is 285-295nm in the HPLC; the mobile phase is acetonitrile-0.02-0.04% phosphoric acid water solution, wherein, the phase A is acetonitrile, and the phase B is 0.02-0.04% phosphoric acid water solution; the analysis time is 50 min; gradient elution.
6. The method for detecting the main effective components of cinnamon and storax in the pharmaceutical preparation according to claim 5, wherein the specific procedure of gradient elution is as follows: 0-25 min, phase A: the volume ratio of the phase B is 20: 80-40: 60, adding a solvent to the mixture; 25-35 min, phase A: the volume ratio of the phase B is 40: 60-50: 50; 35-40 min, phase A: the volume ratio of the phase B is 50: 50-100: 0; 40-50 min, phase A: the volume ratio of the phase B is 100: 0-100: 0.
7. the method for detecting the main effective components of cinnamon and storax in the pharmaceutical preparation according to claim 3, wherein in step 3), the HPLC comprises the following detection conditions: the detector is a photodiode array detector; the chromatographic column is a T3 chromatographic column; the column temperature is 25-35 ℃; the flow rate is 0.8-1.2 ml/min; the sample injection amount is 5-15 mul.
8. The use of the method according to any one of claims 1 to 7 for detecting the major active ingredients of cinnamon and storax in a pharmaceutical preparation for quality detection of cinnamon and storax in a pharmaceutical preparation.
9. A quality detection method for cinnamon and storax in a pharmaceutical preparation comprises that the cinnamon and storax in the pharmaceutical preparation are more than or equal to 0.092% in terms of cinnamic acid.
10. The method for detecting the quality of cinnamon and storax in a pharmaceutical preparation according to claim 9, wherein the weight of the pharmaceutical preparation is 22.5mg per pill, and the content of cinnamon and storax in each pill is more than or equal to 21 μ g in terms of cinnamic acid.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2020112966954 | 2020-11-18 | ||
| CN202011296695 | 2020-11-18 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113237961A true CN113237961A (en) | 2021-08-10 |
Family
ID=77129993
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110043810.5A Pending CN113237961A (en) | 2020-11-18 | 2021-01-13 | Method for detecting main drug effect components of cinnamon and storax in pharmaceutical preparation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113237961A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115267005A (en) * | 2022-08-09 | 2022-11-01 | 上海和黄药业有限公司 | Method for detecting content of multi-index components in Kaixiong powder |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2659442A1 (en) * | 2006-10-18 | 2008-07-31 | Jiangsu Kanion Pharmaceuticals, Co. Ltd. | Cinnamomi and poria composition and uses thereof |
| WO2017148418A1 (en) * | 2016-03-03 | 2017-09-08 | 石家庄以岭药业股份有限公司 | Method for determining component contents of chinese medicine composition |
| CN108802233A (en) * | 2018-07-03 | 2018-11-13 | 上海和黄药业有限公司 | The detection method of a variety of effective components in a kind of Chinese cassia tree |
| CN111398505A (en) * | 2020-04-28 | 2020-07-10 | 健民药业集团股份有限公司 | Method for simultaneously detecting contents of five components of traditional Chinese medicine for treating infantile enuresis |
-
2021
- 2021-01-13 CN CN202110043810.5A patent/CN113237961A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2659442A1 (en) * | 2006-10-18 | 2008-07-31 | Jiangsu Kanion Pharmaceuticals, Co. Ltd. | Cinnamomi and poria composition and uses thereof |
| WO2017148418A1 (en) * | 2016-03-03 | 2017-09-08 | 石家庄以岭药业股份有限公司 | Method for determining component contents of chinese medicine composition |
| CN108802233A (en) * | 2018-07-03 | 2018-11-13 | 上海和黄药业有限公司 | The detection method of a variety of effective components in a kind of Chinese cassia tree |
| CN111398505A (en) * | 2020-04-28 | 2020-07-10 | 健民药业集团股份有限公司 | Method for simultaneously detecting contents of five components of traditional Chinese medicine for treating infantile enuresis |
Non-Patent Citations (3)
| Title |
|---|
| 李洪芹: "HPLC法同时测定独红骨刺丸中的3种成分", 《海峡药学》 * |
| 李静 等: "HPLC法测定养心定悸口服液中肉桂酸的含量", 《实用医技杂志》 * |
| 郝福 等: "反相高效液相色谱法测定苏合香丸中肉桂酸的含量", 《中国现代应用药学》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115267005A (en) * | 2022-08-09 | 2022-11-01 | 上海和黄药业有限公司 | Method for detecting content of multi-index components in Kaixiong powder |
| CN115267005B (en) * | 2022-08-09 | 2023-12-26 | 上海和黄药业有限公司 | Method for detecting content of multi-index components in yippee powder |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110376294B (en) | Method for constructing fingerprint spectrum of snakegourd fruit formula particles | |
| CN113960203B (en) | Method for detecting characteristic spectrum of tianlong formula particles | |
| CN110780007A (en) | Method for evaluating 6 component contents of mango cough relieving tablet by HPLC (high performance liquid chromatography) method | |
| CN113237961A (en) | Method for detecting main drug effect components of cinnamon and storax in pharmaceutical preparation | |
| CN110133158B (en) | HPLC fingerprint detection method of wine steamed coptis chinensis | |
| CN111272941A (en) | Ultra-high performance liquid chromatography determination method for bear gall powder in phlegm heat clearing injection | |
| CN113237960A (en) | Method for detecting main drug effect components of calculus bovis factitius in pharmaceutical preparation | |
| CN114184719B (en) | Dual-wavelength fingerprint spectrum establishment method of Baoyuan decoction and standard fingerprint spectrum thereof | |
| CN112611823A (en) | Method for detecting main active ingredients of borneol and artificial musk in medicinal preparation | |
| CN115524424A (en) | Capsella bursa-pastoris sample quality control method | |
| CN108802233A (en) | The detection method of a variety of effective components in a kind of Chinese cassia tree | |
| CN111896637B (en) | Detection method of Jinqing intermediate and fingerprint spectrum construction method thereof | |
| CN110031577B (en) | Quality detection method and identification application of traditional Chinese medicine or traditional Chinese medicine composition preparation | |
| CN114755338A (en) | Detection method of agastache rugosus and preparation thereof | |
| CN113655165A (en) | Fingerprint spectrum detection method of postpartum rehabilitation ointment | |
| CN111505156A (en) | Fingerprint spectrogram quality determination method for herba Cirsii formulation granules | |
| CN111413429A (en) | Detection method of gardenia intermediate and fingerprint spectrum construction method thereof | |
| CN110118841A (en) | A kind of construction method of the HPLC characteristic spectrum of oral liquid for clearing liver and gallbladder | |
| CN115420830B (en) | GC-MS method for quantitatively detecting volatile components of golden bell oral liquid | |
| CN114755344B (en) | Method for determining dicaffeoylquinic acid content in Monascus purpureus extract | |
| CN113533598B (en) | High performance liquid chromatography analysis method for content of scutellaria baicalensis in infantile lung heat cough and asthma granules | |
| AU2021106162A4 (en) | High-Performance Liquid Chromatography (HPLC) Characteristic Chromatogram of Huatuo Zaizao Pill, and Construction Method and Application thereof | |
| CN113791147B (en) | Quality detection method of cudrania root | |
| CN113820405A (en) | Quality control method for pyrrosia lingua formula granules | |
| CN110873774B (en) | Method for establishing high performance liquid chromatography fingerprint spectrum of cough-relieving immediate effect pills |
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 | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20210810 |
|
| RJ01 | Rejection of invention patent application after publication |